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COOOOO <IM I - 9 ^ W Tennant Farm Herd Health Investigation _> r i cS ' 1. 2 <+ Cattle Team Report i?.Stahl Tennant Farm Herd Health Investigation Cattle Team Report r: :f EPA/DuPont Cattle Team Dr. Perry L. Habecker Dr. Lisa A. Davis-Heller Dr. Peter G. .Vloisan Dr. Robert J. Munson Dr. Robert H. Poppenga Dr. Greg P. Sykes u> V* CD - o m o CO -- i r n X o m rK cO ro December 23. 1999 000004 EID151682 R G S000885 Tennant Farm Herd Health Investigation TABLE OF CONTENTS Section 1.0. 1.1. 2.0. SUMMARY Signatures INTRODUCTION 3.0. 3.1. 3.1.1. 3.1.2. 3.2. 3.2.1. 3.2.2. 3.2.3. 3.2.4. 3.2.5. METHODS Cattle Team members meetings Animal Data videotapes diagnostic pathology reports April 1999 site visit data miscellaneous data wildlife 4.0. 4.1. 4.1.1. 4.1.2. 4.1.3. 4.2 4.3. 4.3.1. 4.3.2. 4.3.3. RESULTS Cattle videotapes diagnostic pathology reports April 1999 site visit data a. herd historv (4/8/99 interview) b. physical examination of cattle (4/7/99) - age. pregnancy status - body condition scores, girth - Clinical signs c. clinical pathology - hematology - clinical chemistry - special chemistry - serology - parasitologv d. hay and grain analysis Miscellaneous Data Wildlife videotapes small mammal data deer studies 5.0. DISCUSSION - endophyte toxicity - pinkeye - malnutrition Cattle Team Repon Page 4 5 6 6 6 6 7 7 7 8 8 11 12 12 12 12 14 15 15 16 16 16 16 16 16 16 17 17 18 18 19 19 19 19 19 20 20 22 23 2 Q00005 EID151683 R G S000886 Tennant Farm Herd Health Investigation Cattle Team Report Section 5.0. TABLE OF CONTENTS (continued) DISCUSSION (continued) - copper deficiency - toxicology issues 6.0. RECOMMENDATIONS 7.0 CONCLUSION 8.0 REFERENCES - ...................................................... - -- 9.0 GRAPHS Graph 1: Individual Prolactin Values 10.0 FIGURES Figures 1and 2: Faceflies Figure 3: Corneal opacity Figure 4: Delayed shedding Figures 5 and 6: Coronitis Figures 7 and 8: Cattle standing in water 11.0 TABLES Table 1: Review of Tennant Farm Videotapes Table 2: Individual Animal Data: Clinical Signs Table 3: IndivL jal Animal Data: Hematology (erythron, platelets) Table 4: Individual Animal Data: Hematology (leukon), and Special Chemistry Table 5: Individual Animal Data: Clinical Chemistry Table 6: Individual Animal Data: Serology and Fecal Exam Table 7: Tennant Farm Grain and Hay: Nutritional Analysis 12.0 APPENDICES Appendix A: Abbreviated Curriculum Vitae of Cattle Team Members Appendix B. Diagnostic Pathology Reports (Ohio Dept. Agric.: #497797; Michigan Animal Health Diagnostic Lab.: #1792571; Univ. Penn. Lab. of Large Animal Pathol, and Toxicol.: #UP9902702, #9901437) Appendix C: Dry Run Safety Plan Appendix D: Figures 1-42 : Photographs of the 42 adult cattle Figures 43-66 : Miscellaneous photographs of the Tennant Farm and cattle. Appendix E: Herd Health History (April 8, 1999 interview) Appendix F: Diet Analysis: Computer Software Simulation Page 23 24 24 26 26 29 30 31 32 33 33 34 35 36 37 44 46 48 50 52 54 55 56 58 70 71 106 109 3 000006 EID151684 R G S000887 Tennant Farm Herd Health Investigation Cattle Team Report 1.0 SUMMARY Introduction: The objective of this investigation was to evaluate the health status of Mr. Earl Tennant's cattle herd and determine possible causes of any problems. The investigation included an examination of relevant historical data as well as the collection and evaluation of new data. The investigation terminated in the production of this report, which outlines study findings and makes recommendations to improve herd health. Methods: The Tennant farm herd health investigation was conducted by a team of six veterinarians with collective experience in bovine diseases, herd health management, toxicology, pathology, and wildlife diseases. In addition to numerous meetings and discussions, the cattle team visited the Tennant Farm on April 7-8, 1999 in order to collect relevant data. The cattle team reviewed historical data (e.g., videotapes, diagnostic laboratory reports) and contemporary data (e.g., clinical examinations, blood tests). Diagnoses and recommendations were based upon the data collected. Results: The multifaceted disease investigation of the adult cattle in the Tennant beef herd revealed health problems that were related to endophyte toxicity, infectious keratoconjunctivitis (pinkeye), malnutrition, and copper deficiency. Clinical and historical data, with prolactin values of some animals, were consistent with endophyte mycotoxicosis. An examination of videotapes made during the summer months and clinical examinations of previously affected adult animals during the herd visit indicated prolonged severe enzootics of facefly {Musca autumncilis) infestation and concomitant pinkeye. Hay analysis, cattle body condition scoring, and an evaluation of the mineral and grain rations fed were consistent with protein-energy malnutrition and macromineral/trace nutrient deficiencies. Earlier laboratory data, clinical signs, and serum testing at the time of the herd visit were indicative of severe copper deficiency in the cattle. Conclusion: There was conclusive evidence that the Tennant cattle herd was. and continues to be. suffering from four major disease entities, some of which were potentially interrelated: endophyte toxicity (fescue mycotoxicosis), pinkeye, malnutrition, and copper deficiency. As substantiated by the clinical and laboratoryfindings. and historical data, these four conditions readily account for the chronic herd health problems on the Tennant farm. The herd health investigation revealed deficiencies in herd management, including poor nutrition, inadequate veterinary care, and lack of fly control. The lack of vaccination and internal parasite control programs did not appear to have a substantial impact on this relatively isolated herd. Despite an exhaustive review of historical and contemporary herd data, there was no evidence of toxicitv associated with chemical contamination of the environment. 000007 4 EID151685 R G S000888 Tennant Farm Herd Health Investigation 1.1 Signatures Cattle Team Report L. Perry(LJflabecker, VMD, Dipl. ACVP /z /g .3 (9 9 Date ] C t Lisa & Davis-Heller, DVM Dat Peter G. Moisan, DVM, Dipl. ACVP, ABVP Date 11 Robert J. Munson, VMD oo iU l Date obert H. Poppenga, DVM, PhD, Dipl. ABVT /-3 -O o Date Greg P. Dipl. ACVP, ABT Date 000008 5 EID151686 R G S000889 Tennant Farm Herd Health Investigation Cattle Team Report 2.0 INTRODUCTION The objective of this investigation was to evaluate the health status of Mr. Earl Tennant's cattle herd and determine possible causes of any problems. The investigation included an examination of relevant historical data as well as the collection and evaluation of new data. The investigation terminated in the production of this report, which outlines study findings and makes recommendations to improve herd health 3.0 METHODS 3.1. Cattle Team 3.1.1. Members The Tennant Farm Herd Health Investigation was assigned to a team of six veterinarians ("cattle team"). The team was constituted to include expertise in bovine diseases, herd health management, toxicology, pathology, and wildlife diseases. Representatives of the DuPont Company (DuPont) selected three members (Drs. Sykes, Davis-Heller, Moisan); representatives of the United States Environmental Protection Agency (EPA) selected three members (Drs. Habecker, Poppenga, Munson). Abbreviated Curriculum Viiae of the cattle team members arc included in this report (Appendix A). Cattle Team Members Associations Perry Habecker. VMD. Dipl. ACVP Chief. Large Animal Pathology. Laboratory of Pathology and Toxicology. Univ. of Penn. School of Veterinary Medicine. N>. ' Bolton Center, Kennett Square. PA. Lisa Davis-Heller. DVM Private Practitioner. St. Mary's Veterinary Clinic. St. Mary's. WV. Peter Moisan. DVM. Dipl. ACVP. Dipl. Veterinary Pathologist, Rollins Animal Disease Diagnostic ABVP Laboratory (state laboratory), Raleigh, NC. Robert Munson. VMD Field Investigator. Center for Animal Health and Productivity. Univ. of Penn. School of Veterinary Medicine. Kennett Square. PA. Robert Poppenga. DVM. PhD. Dipl. ABVT Chief. Toxicology. Laboratory of Pathology and Toxicology, Univ. of Penn. School of Veterinary Medicine. New Bolton Center, Kennett Square. PA. Greg Sykes. VMD. Dipl. ACVP, Dipl. ABT Pathologist, Safety Assessment, DuPont Pharmaceuticals Company, Stine Research Center, Newark, DE. DVM. VMD = Docior of Veterinary Medicine Dipl. ACVP = Diplomate, American College of Veterinary Pathologists Dipl. ABVP = Diplomate. American Board of Veterinary Practice (Food Animal and Beef Cattle Specialist) Dipl. ACVT = Diplomate. American College of Veterinary Toxicologists Dipl. ABT = Diplomate. American Board of Toxicology At the first meeting of the cattle team (March 9. 1999), Perry Habecker was elected Scientific Leader (i.e., chairman) and Greg Sykes was elected Coordinator. The 000009 6 EID151687 R G S000890 Tennant Farm Herd Health Investigation Cattle Team Report chairman and coordinator were instructed to communicate with the "steering committee" leaders, Drs. Sarah Caspar (EPA, Region III) and Ralph Stahl (DuPont Corporate Remediation Group), as needed. This "steering committee" included Drs. Caspar, Stahl, Mike Horne (USFWS), Mark Sprenger (EPA-OERR), and Rudy Valentine (DuPont Specialities Chemicals toxicologist). 3.1.2. Cattle Team Meetings The cattle team met formally on four occasions, including the Tennant Farm site visit. Cattle Team Meetings Date Location March 9. 1999 New Bolton Center, Kenne Square, PA March 23. 1999 New Bolton Center, Kenne Square, PA April 7-8, 1999 Tennant Farm, Wood County, WV* July 28-29. 1999 New Bolton Center, Kenne Square, PA * including pre-meeting planning session at Holiday Inn. Parkersburg. WV In addition, several in-person discussions took place at New' Bolton Center between and among Drs. Habecker, Munson, Poppenga, and Sykes between March 1999 and the issuance of this report. All cattle team members utilized the telephone, internet email, and US postal systems for pair and group discussions and information sharing. 3.2. Animal Data 3.2.1. Videotapes Two videotapes were supplied to the cattle team by the steering committee. Each of the tapes was a collection of videotapes made, and narrated, by Mr. Earl Tennant on his farm and the adjacent landfill area. Although the tapes were edited and include material from different seasons, it appeared that these tapes were all produced between 1995 and 1997. Tennant Farm Videotapes Tape Title #1 "Tennant Farm: New England. Wood County. WVa - January and February, 1997" #2 "Dry Run: Harris PC. Wood Co.; Off North Fork of Lee Creek. New England. WVa" Animal Case Numbers 1 - 18 19-60 Both of these tapes were viewed, in their entirety, by the cattle team members individually. They were also reviewed at a cattle team meeting on July 28,h (Dr. DavisHeller not present). A total of 60 animal cases, ranging from a dead crayfish to groups of sick cows, were evaluated. Items on the tapes that were not animal related (e.g., water treatment) were noted but not evaluated. Table d is a compilation of the relevant animal health data derived from these tapes. This table does not include many assertions and OOOOIO 7 EID151688 R G S000891 Tennant Farm Herd Health Investigation Cattle Team Report interpretations, made by the narrator in the tapes, which the team members considered to be incomplete or erroneous. 3.2.2. Diagnostic Pathology Reports Two pathology reports (Appendix B) were supplied to the cattle team by the steering committee. Both were issued by state animal diagnostic laboratories in response to the submission of dead animals or tissues from the Tennant Farm. These reports were reviewed by the cattle team. A third pathology report (Appendix B) was produced subsequent to the elective sacrifice of one animal (# 37) on June 10, 1999. The cattle committee decided, at a meeting on May 28, 1999, that it would be informative to have a toxicology screen on tissues from one or two older cows. Mr. Tennant was asked to select one or two of the cattle that he considered to be in the worst condition. He chose to sacrifice cow # 37, a 7-year old red cow that had freshened a few months earlier. This animal was euthanized by gunshot by Mr. Tennant on the farm on June 10, 1999. Mr. Tennant performed the dissection in the presence of Dr. Davis-Heller. Tissues were collected (Dr. Davis-Heller) for histopathologv and analytical toxicology. Individual Animal Pathology Reports Pathology Report Date Name of Diagnostic Laboratory March 10, 1997 Animal Disease Diagnostic Laboratory. Ohio Department of Agriculture. Reynoldsburg. OH March 12. 1997 Animal Health Diagnostic Laboratory. College of Veterinary Medicine. Michigan State University. Lansing MI July 5, 1999 Laboratory of Large Animal Pathology and Toxicology. New Bolton Center. University of - - -- --i Pennsvlvania. Kennett Square, PA Case Material (case number) dead 6-month-old bull calf (#4977-97) tissues 4-year-old Holstein cow (#1792571) tissues 9-year-old Holstein cow C4 79257!) tissues 7-vear-old cow (#UP9902702: UP9901437) 3.2.3. April 1999 Site Visit Data The cattle team visited the Tennant Farm and adjacent Dry Run landfill site on April 7 and 8, 1999 in order to collect data and biological samples relevant to the herd health investigation. Before proceeding to the site on April 7th, a brief safety meeting was held at the Parkersburg Holiday Inn during which the Dry Run Safety Plan (Appendix C) was reviewed. The 6 cattle team members, 2 of the steering committee members (Caspar, Horne) and 2 USFWS employees were present. April 7. 1999 The cattle team had planned to inspect the farm and adjacent landfill, view the herd grazing, and interview Mr. Tennant on the afternoon of Wednesday April 7lh. However, 8 OOOOll EID151689 R G S000892 Tennant Farm Herd Health Investigation Cattle Team Report due to a misunderstanding, Mr. Tennant had corralled his entire herd on Wednesday morning. When the cattle team and EPA/USFWS personnel arrived at approximately 3:15 PM, the herd had been contained in a small enclosure, without food or water, for several hours. The team therefore elected to conduct the individual animal examinations and specimen collections immediately. A new head-hold restraint device had been installed next to the enclosure to facilitate animal handling. Mr. Earl Tennant and his brother were very helpful in handling the cattle and moving them into the restraint device. The 41 adult cattle were individually run into the chute, secured with the head hold and a halter, and examined. Each adult (38 cows, 2 bulls, and 1 steer) was given a general physical examination including observation of gross abnormalities, grading of body condition, and estimation of age based on the incisors. Blood was collected by jugular venipuncture (2 red top, 1 green top, and 1 lavender top tubes) from each cow and fecal samples were rectally collected from each adult. Cows were rectally palpated to determine their pregnancy status. All examined adult animals received two ear tags, one with an identification number and one with an impregnated insecticide. One Hereford cow (i.e.. the 39thcow. 42nd adult) escaped the chute prior to examination, aging, uterine palpation, blood collection, and ear tagging. Each adult bovine was individually photographed and identified (Appendix D, photographs 1-42). Only 1 cow (animal #22) received treatment - a mass, most consistent w'ith a dermoid cyst, was incised and drained (Appendix D, photographs 49-51). Additional photographs of the farm (Appendix D, photographs 43, 54-58), individual animal "lesions" observed on April 7, 1999 (Appendix D, photographs 4 ' 48, 52, 53). photographs supplied to the team by Mr. Tennant (Appendix D, photographs 60-65). and a photograph of cattle team and steering committee members (Appendix D, photograph 66) are also included in this report. Adult Cattle Procedures physical examination body condition scored ase estimation rectal prepnancv examination pirth measurement fecal sample collection blood collection ear tapped: animal identification ear tapped: insecticide photopraphed Twelve calves were examined as a group. The smaller seven received insecticidal ear tags; the larger five received no ear tags as Mr. Tennant indicated that they were to be sold in the very near future. Although the cattle team was prepared to do a post-mortem examination on one or more cattle, the team decided that there was no animal sufficiently ill to justify euthanasia and necropsy at the time of the visit. 9 000012 EID151690 R G S000893 Tennant Farn. Herd Health Investigation Cattle Team Report All animal biological samples (blood and feces) were taken back to New Bolton Center by Dr. Habecker. Blood samples were subjected to hematology, serum chemistry, and serology. Randomly selected fecal samples were subjected to fecal flotation. The following clinical pathology parameters were evaluated: Hematology leukoevte (total and differential) count ` hematocrit ervthrocyte count ` mean corpuscular hemoglobin hemoglobin concentration ` mean corpuscular hemoglobin concentration mean corpuscular volume ` red cell distribution width platelet count ` platelet distribution width mean platelet volume `These parameters were calculated using the measured data. Serum Chemistry aspartate aminotransferase potassium gamma glutamvl transferase calcium creatine kinase chloride blood urea nitrogen magnesium creatinine phosphorus total protein copper albumin selenium ` globulin pepsinogen sodium " prolactin * globulin was calculated from the measured albumin and total protein data. " prolactin was measured from blood plasma derived from EDTA tubes (lavender top). Serology bovine leukemia virus (et epizootic hemorrhagic disease of deer (agid) bovine virus diarrhea virus (sn) M \c o b a c te r iu m p a ra tu b e rc u lo sis (Johne's) (e) bovine virus diarrhea virus (mpi leptospirosis (sn> blue tongue virus (agidl brucellosis (sn) (cl = elisa test: (sni = serum neutralization test: (mpt = microplate assay; (agid) = agarose gel immunodiffusion test April 8. 1999 Three members of the cattle team (Drs. Moisan, Munson, and Davis-Heller) met with Mr. Earl Tennant and recorded his recollection of the 1998-1999 herd health history (Appendix E). Drs. Caspar and Home gave all of the cattle team members a driving tour of the property between the Tennant barn and the Dry Run landfill, including a drive within the landfill. During those two trips, the team members were able to see the connecting pastures. 000013 10 EID151691 R G S000894 Tennant Farm Herd Health Investigation Cattle Team Report adjacent creeks, neighboring cattle grazing, and wildlife. The landfill was open and being utilized at the time of the site visit. While on site, the cattle team made general observations regarding the environment, including the flora, fauna, and man-made structures. Core samples of three large round hay bales were collected (3 samples/bale) from bales in the bam yard. Mr. Tennant gave the team some receipts (dates ranged from 2/98 to 12/98) from complete, mixed feed, and mineral supplement he had purchased. All plant biological samples (bailed hay core samples and grain sample) were subjected to nutritional analysis by the Forage Testing Laboratory of the North Carolina Department of Agriculture (Table 7). These data were used in an Excel spreadsheet program (CNCPS v 3.1) from Cornell University to evaluate the diets in two cow models under two feeding situations (Appendix F, models 1-4). The following nutritional parameters were evaluated: Hay and Grain Analysis drv matter crude protein unavailable protein adjusted crude protein acid determent fiber total digestible nitrogen NE (lactation) calcium phosphorus sodium magnesium sulphur potassium copper iron manganese zinc nitrate ion selenium 3.2.4. Miscellaneous Data During the site visit in April. Mr. Tennant was asked to notify Dr. Lisa Davis-Heller or Dr. Sarah Caspar if there were any future signs of disease problems in his herd. The team wished to collect whatever data might be relevant to their assessment of the health status of the Tennant herd. Between April 8 and December 23, 1999, Mr. Tennant consulted twice with Dr. DavisHeller. Subsequent to notifying Dr. Caspar of a newborn calf which was born with a "cloudy eye", Mr. Tennant brought the calf to Dr. Davis-Heller's clinic (April 21, 1999). Dr. Davis-Heller examined the calf and made a diagnosis. Mr. Tennant left with the calf. On May 26, 1999, Dr. Davis-Heller examined cow #30 for a lump under the right mandible. 000014 11 EID151692 R G S000895 Tennant Farm Herd Health Investigation Cattle Team Report 3.2.5. Wildlife EPA and DuPont each sponsored environmental studies that included the trapping of small mammals in the grazing fields of Mr. Tennant's cattle. These reports were supplied to the cattle team by the steering committee. The data included in these reports, as it relates to small mammal pathology, were evaluated by the cattle team. Small Mammal Studies Report Date Report Title November. 1997 Dry Run Creek: Washington, Wood County, West (draft) Virginia. USEPA Environmental Response Team December 3. 1998 (letter i Small Mammal Trapping Effort; Dry Run Landfill, Washington. WV (URS Greiner Woodward Clyde study) Animal Data Section Table 39: Results of Histopathology for the Meadow Vole Table 1: Data Summary, Small Mammal investigation The cattle team was also made aware of periodic deer reproduction surveys conducted by Dr. Crum and associates of the West Virginia Department of Natural Resources (WV DNR) in the Dry Run area, however no reports were available. 4.0 RESULTS 4.1. Cattle 4.1.1. Videotapes (Table 1) Two videotapes of several hours duration were reviewed. Forty cattle scenes and 20 wildlife scenes were presented, depicting numerous lesions. The individual scenes (i.e.. cases), along with the cattle team's diagnosis/comment, are presented in Table 1 in the order in which they were presented in the videotapes. The herd health problems identified in these videos were summarized as follows: 1. Keratitis: Corneal ulcers, scars and opacities, as well as blepharospasm, were observed in many cattle. These were all considered to be manifestations of "pinkeye", an infectious keratoconjunctivitis caused by bacteria such as Morexella bovis. The facefly (Musca autunmalis) problem observed in these videos and the lack of effective fly control were consistent with the diagnosis of severe "pinkeye" in these cattle. Affected young and adult cattle were observed to have 2-3 to 25-50 or more faceflies feeding at the medial canthus of each eye and on the lacrimal secretions. Figures 1 and 2 are prints from videotape #2, illustrating the facefly problem on a calf and her dam, respectively. Both of these animals were blind from chronic keratoconjunctivitis. Figure 3 (videotape #2) demonstrates an early central corneal ulcer on the left eye of a calf; faceflies are seen by the lower eyelid. 000015 12 EID151693 R G S000896 Tennant Farm Herd Health Investigation Cattle Team Report 2. Hair loss, neck and tail: The cervical alopecia was most consistent with a lice problem. The alopecia observed on the tails of some cattle (e.g., loss of "switch") may also have been due to lice, or may have been related to fescue mycotoxicity. 3. Poor shedding of hair coats; This clinical sign, most prominent in the late spring and summer segments of the videotapes, is consistent with multiple nutritional deficiencies as well as fescue mycotoxicosis. Figure 4 demonstrates a cow from videotape #2 that was described by Mr. Tennant as having delayed shedding of the winter coat. 4. Hair depigmentation: The discoloration of the cattle hair coats is a change most often associated with a nutritional deficiency, and is especially common in cases of copper deficiency. 5. Coronitis: Alopecia and erythema above the coronary band may be seen with fescue mycotoxicosis, mechanical trauma, or as a non-specific dermatitis. Figures 5 and 6 (videotape #2) are representative of the erythematous foot lesions observed by Mr. Tennant in many of his cattle. Figures 7 and 8 (videotape #2)) demonstrate the cattle's preference for standing in the water when these lesions were present. Fescue mycotoxicosis ("fescue foot"-) was considered the most likely diagnosis of this condition. 6. "Hunched up" or painful stance: This is a clinical sign often associated with abdominal or foot pain, but has been specifically described as a manifestation of the fescue endophyte toxicity complex. 7. Thin body condition: Poor body condition is a non-specific finding and the cases presented most likely represented different etiologies. The emaciated dead calf with serous atrophy appears to have starved while others appear to have had diarrhea or mastication problems. Considering the other findings in this herd (e.g., feed analysis), nutritional insufficiency (protein-energy malnutrition) was also a likely contributing factor to poor body condition. Some findings described in these tapes were difficult to evaluate by video although they probably were real. For example, the described "lumps" in a cow udder (case # 54) was difficult to see but would be considered an incidental finding in any case. A few cattle were described as "hunched-up" or "humped-up" (cases # 1, 5, J7) - these may have been related to lameness, hardware disease (traumatic reticulopericarditis), fescue mycotoxicosis, or other condition. An individual cow that was slobbering, losing cud, and urinating (case # 13) may also have had hardware disease, although the diagnosis was not clear. Another individual cow (case # 45) which was panting was consistent with fescue mycotoxicosis (i.e., "summer fescue" hyperthermia), although this presentation was certainly non-specific. 000016 13 EID151694 R G S000897 Tennant Farm Herd Health Investigation Cattle Team Report 4.1.2. Diagnostic Pathology Reports (Appendix B) Only four of Mr. Tennant's cattle were submitted for post-mortem diagnostic evaluation of some type. The most significant finding was moderate to marked copper deficiency in the three cows analyzed for heavy metals. Keratitis was observed in the only animal that was formally necropsied. This 6-month old bull calf apparently died from winter starvation (i.e., negative energy balance or protein-energy malnutrition) complicated by untreated intestinal coccidiosis. These reports are included in Appendix B; summaries are presented below: a. Dead 6-month old bull calf submitted to the Animal Disease Diagnostic Laboratory, Ohio Department of Agriculture, Reynoldsburg, OH on 2/27/97; Laboratory report dated 3-10-97 (Appendix B) History: Gross diagnosis: Histopathologic diagnosis: 6-month old bull calf; underweight; diarrhea for one month; received no medication; corneal opacities; died. Emaciation with serous atrophy of fat Intestinal parasitism (trichuriasis) Enteric coccidiosis Keratitis, moderate, chronic, focally extensive b. Tissues from a 4-year old Holstein cow submitted to the Animal Health Diagnostic Laboratory. College of Veterinary Medicine, Michigan State University, Lansing MI on 3/4/97; Laboratory report dated 3-12-97 (Appendix B) History: 4-year old Holstein cow; died 2-18-97; dissected by owner; tissues submitted by EPA Heavy metal screen: Copper deficiency in liver (1.9 ppm vs. 25-150 reference range) and kidney (2.37 ppm vs. 4 - 6 ppm reference range). Manganese was marginally low in liver; cadmium was slightly increased in the kidney. No heavy metals found in urine (urine fluoride was 6.6 ug/ml)[reference range: toxic if > 14 jig/mL] c. Tissues from a 9-year old Holstein cow submitted to the Animal Health Diagnostic Laboratory, College of Veterinary Medicine, Michigan State University, Lansing MI on 3/4/97; Laboratory report dated 3-12-97 (Appendix B) History: 9-year old Holstein cow; died 3-2-97; dissected by owner; tissues submitted by EPA 000017 ta EID151695 R G S000898 Tennant Farm Herd Health Investigation Cattle Team Report Histopathologic diagnosis: (only heart, liver, and kidneys submitted: autolysis and freeze/thaw artifact) Myocardial sarcocysts Heavy metal screen: Copper deficiency in liver (2.03 ppm vs. 25-150 reference range) and kidney (2.33 ppm vs. 4 - 6 ppm reference range). Manganese was marginally low in liver; iron was elevated in the liver; cadmium was slightly increased in the kidney. Urine heavy metals were normal (urine fluoride was 5.55 ug/ml)[reference range: toxic if > 14 pg/mL] Clinical pathology: No significant findings d. Tissues from a 7-year old red cow (euthanized on 6/10/99 and dissected by Mr. Tennant) submitted to the Laboratory of Large Animal Pathology and Toxicology, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA on 6/27/99; Laboratory report dated 7-5-99 (Appendix B) History: 7-year old red cow; euthanized 6-10-99; dissected by owner; tissues submitted by Dr. Davis-Heller. Histopathologic diagnosis: Enteric lesions of minimal significance (forestomach abscesses; intestinal coccidiosis; myocardial sarcocysts). Heavy metal screen: Copper deficiency in liver (7.71 ppm vs. 25-150 reference range). 4.1,3. April 1999 Site Visit Data a. Herd History (Appendix E) The herd history (Appendix E) was based on Mr. Tennant's recollections during the interview on April 8, 1999. A written record of herd health was conspicuously absent. Much of this can be attributed to the lack of outside intervention. There is no record of veterinary care or consultation with an animal nutritionist. Minimal medications have been used on these animals, and there has been no use of vaccines or modern dewormers. Except for the few' animals noted in this document (see Diagnostic Pathology Reports, above), no diagnostic laboratory tests were done on dead animals. The level of cattle herd management has not changed since the installation of the landfill. 000018 15 EID151696 R G S000899 Tennant Farm Herd Health Investigation Cattle Team Report b. Physical Examination (Table 2) Age. Pregnancy Status On April 7. 1999, the cattle team examined 41 animals, including 38 adult cows, 2 adult bulls, and 1 adult steer (Table 2). Estimated ages for the adult cows ranged from 4 to greater than 9 years. This is a herd of aged cattle; 32 of the 38 cows were estimated to be older that 9 years. Twenty-seven (27/38) cows were pregnant. Body Condition Scores Body condition scores (BCS) were based on a 1-9 scale. Scores ranged from 2 to 7; the average score was 3.5 (3=thin, 4=borderline, 5-7 is considered optimum). Clinical Signs Haircoat abnormalities were evident in 9 of the 41 cattle. One animal had an epidermal inclusion cyst that was lanced and emptied. Mammary gland lumps, consistent with chronic mastitis, were interpreted as abscesses or accumulations of fibrous connective (scar) tissue. Rectal examination of one cow suggested the presence of intra-abdominal fat necrosis. c. Clinical Pathology (Tables 3 - 6 ; Graph 1) Hematology Erythron (Table 3): Red blood cell indices were generally in the low-normal range. No animals were considered anemic. Leukon (Table 4): The total white blood cell counts were within the normal range. Differential cell counts were considered invalid due to the 36 hour delay between collection and testing. Clinical chemistry (Table 5) Most electrolyte values were in the high-normal to slightly elevated range, consistent with mild dehydration. Since the cattle were penned during the daylight hours of April 7, which was unseasonably warm, and had no access to water, clinical dehydration was not unexpected. Dehydration is also the most likely explanation for the elevated creatinine in 40/41 cattle. Most (27/41) of the cattle had elevated total protein, which also correlates highly with a diagnosis of dehydration (total protein = globulin fraction + albumin fraction). In all cattle (41/41) the globulin fraction was elevated while the albumin fraction was within the reference range. 000019 16 EID151697 R G S000900 Tennant Farm Herd Health Investigation Cattle Team Report Gamma glutamyltransferase (GGT), a sensitive indicator of active liver disease, was consistently within the normal reference range. Creatine kinase (CK), an indicator of muscle necrosis, was also normal in all of the cattle. However, aspartate aminotransferase (AST), a less sensitive indicator of liver and muscle degeneration, was minimally elevated in 33/41 cattle. This might be from leukocyte dissolution, the result of the 36 hour interval between blood collection and testing. Special chemistry Prolactin: Plasma prolactin levels (Table 4 and Graph 1) were significantly depressed. According to the testing laboratory (Dr. Neal Schrick, University of Tennessee), an average prolactin level for cattle on fescue-free pasture (for April) was 171.6 micrograms/liter (ng/mL) (i.e., reference mean = 171.6 ng/mL). The average of the 41 Tennant samples was 110.1 ng/mL and 36/41 of the Tennant cattle were below the reference mean (171.6 ng/mL). In contrast, an average prolactin level for cattle on endophyte-infected fescue (simulated by ergotamine tartrate administration) was 105.8 ng/mL (data from Dr. Schrick). This average was similar to the Tennant herd average (110.1 ng/mL) and 24/41 of the cattle were below this reference level (105.8 ng/mL). According to Dr. Schrick, these findings were highly supportive of endophyte toxicity in the Tennant herd. Copper: Blood copper levels were in the deficient range for 26 of 41 animals; one copper assay was below the limit of detection. Selenium: Blood selenium levels were normal for 41 of 41 cattle. Pepsinogen: Pepsinogen, a blood enzyme indicator of parasite damage to the abomasum, was measured in 10 cows, one steer and one bull. All values were within the normal reference range. These data supported the conclusion that abomasal ostertagiasis was not a problem in adult cattle in this herd. Serology (Table 6) BLV (Bovine Leukemia Virus) Johne's (Mycobacterium paraiuberculosis antigen) Brucella BVD (Bovine Virus Diarrhea antigen) BVD Microplate Assay (viremia detection) Bluetongue Leptospira interrogans (5 subvarieties) EHD type 2 (Epizootic Hemorrhagic Disease of deer) 1/41 positive 1/14 positive 0/41 2/41 positive 0/41 0/41 3/41 positive 2/12 positive None of the serology values suggested a herd-wide problem. Positive titers for BVD and Leptospira might reflect residual vaccination titers in purchased animals or natural exposure to these diseases. The discovery of two positive EHD titers was evidence that this disease has been in the local deer population. 000020 17 EID151698 R G S000901 Tennant Farm Herd Health Investigation Cattle Team Report Parasitology Routine fecal flotation (Table 4) revealed parasite ova in 11 of 14 randomly selected fecal samples. The magnitude of ova shedding could be categorized as rare to moderate. Parasite groups included coccidia, tapeworms, and strongyle-type nematodes. Intestinal parasitism was not considered to be a major herd problem in the adult cattle. The shedding of small numbers of oocysts and helminth ova is expected of mature cattle. d. Hay and Grain Analysis (Table 7) During the cattle team's visit to the Tennant farm (4/8/99), Mr. Tennant was interviewed regarding herd feeding practices. Three large round bales of hay in the barnyard, identified as representative of the herd's forage, were visually inspected and core samples (3/bale) were collected for analysis. This forage (i.e., hay, grass/fescue) was fed ad libitum. During the site visit, forage quality was subjectively evaluated as poor. The fiber level of the hay appeared to be high and forage analysis (Table 7) later substantiated that observation. Fiber levels of forage play an important role in determining how much of a feed can be consumed. Dry matter intake (DMI) is inversely related to fiber levels. According to Mr. Tennant, mature animals, on the Tennant farm, were also fed five pounds per head per day of a grain mix composed of dry ear com, soybean meal, and minerals (rate could not be validated, despite the existence of grain receipts). A sample of this feed was taken and analyzed. An Excel spreadsheet program (CNCPS v 3.1) from Cornell University was used to evaluate the diets. Using the forage and grain nutritional analysis conducted by the Forage Testing Laboratory of the North Carolina Department of Agriculture (Table 7), two models for mature cows (early lactation and dry pregnant) were selected to examine the current feeding program (Appendix F). According to the computer models, a cow in early lactation (model #1) in the spring of 1999 fed five pounds of this grain and free choice hay was in severe negative energy balance (72.5% of requirement) and less severe protein deficiency (90.9% of requirement). The limited nutrients available would have a negative impact on milk production, persistency of production, and calf growth. A cow under this feeding system would require 2.4 times the corn currently being fed to be isocaloric (model #2). Similarly, a dry pregnant cow (model #3) in the same feeding system would also be in negative energy balance (75.2% of requirement) but not deficient in protein (125%). The same dry pregnant cow maximizing dry matter intake (model #4) would still be in negative energy balance (87.7%). 000021 18 EID151699 R G S000902 Tennant Farm Herd Health Investigation Cattle Team Report The body condition scores (BCSs) of the herd were consistent with the energy deficit of the diets. The impact of the poor quality forage on this farm and subsequent energy deficits were considered to have chronic effects well after the cows return to adequate pasture. 4.2. Miscellaneous Data Following the site visit, Mr. Tennant consulted with Dr. Davis-Heller regarding a suspected sick animal. Mr. Tennant brought a calf with a "cloudy eye" to Dr. DavisHeller's clinic (April 21, 1999). Dr. Davis-Heller examined the calf and determined that there was nothing wrong with the eye, except for a slight mucoid exudate on the cornea which was easily wiped away. On May 26, 1999, Dr. Davis-Heller examined cow #30 for a lump under the right mandible. The lesion was aspirated (pus) and subsequently lanced. 4.3. Wildlife 4.3.1. Videotapes (Table 1) The 20 wildlife cases presented in the videotapes were all dead and, except for one case (case # 49), of no diagnostic value. These were all considered to be incidental deaths since there did not appear to be any consistency in the species, location, or timing of the deaths. In fact, these deaths appeared to be most consistent with the random wildlife carcasses which would be found in a healthy ecosystem. The individual dead deer (case # 49) which presented with hemorrhage from the nostrils may have died from epizootic hemorrhagic disease (EHD). This diagnosis would correspond with the known existence of this disease in local deer (personal communication: Dr. Crum to Dr. Sykes) and the finding of some EHD seropositive cattle in the Tennant herd. 4.3.2. Small Mammal Data Two pathologists reviewed the liver and kidney histology from 45 small rodents (voles, shrews, and mice) captured on the Tennant property in 1997 (USEPA Environmental Response Team draft report: Dry Run Creek, Washington, Wood County, West Virginia; Table 39). No lesions characteristic of hepatotoxicitv or nephrotoxicity were evident. Although several anomalies were identified, none were considered suggestive of toxic teratogenesis. The 1998 small mammal trapping (URS Greiner Woodward Clyde study, December 3, 1998) observed no gross abnormalities in the captured animals (voles, shrews, and mice). 4.3.3. Deer Studies The cattle team was made aware of the periodic deer reproduction surveys conducted by the West Virginia Department of Natural Resources (WV DNR) in the Dry Run area. Although the team reviewed some of the data sheets from these studies, no reports were available. One team member (Sykes) had a telephone conversation with Dr. Crum of the 000022 EID151700 R G S000903 Tennant Farm Herd Health Investigation Cattle Team Report WV DNR regarding deer herd health in the Dry Run area in order to acquire some additional information. From this conversation, it was the cattle team's understanding that: - evidence of low fertility rates in young does was probably attributable to deer overpopulation; - epizootic hemorrhagic disease (EHD) has been diagnosed clinically and serologically in dead and captured deer, respectively; - bovine virus diarrhea (BVD) has been diagnosed serologically in deer not far from Dry Run; In addition, the team has learned that outbreaks of EHD in West Virginia deer were reported between 1980 and 1989 [Southeastern Cooperative Wildlife Disease Study publication, Field Manual of Wildlife Diseases in the Southeastern U.S., 2nd ed.J. Since there was no evidence of an EHD or BVD outbreak in the Tennant herd, the deer population was not considered to be a contributing factor in the cattle herd health problems. Also, there was no evidence that the deer herd was a useful sentinel species for any of the disease entities identified in the Tennant herd. 5.0 DISCUSSION The six veterinarians comprising the cattle team agreed that there was conclusive evidence that the Tennant herd was suffering from four major disease entities: endophyte toxicity, pinkeye, malnutrition, and copper deficiency. The clinical, laboratory, and historical data substantiate that these four conditions can readily account for the chronic herd health problems on the Tennant farm. Endophyte toxicity According to the history acquired on April 8, 1999, the herd has been fed a diet of KY31 fescue hay in the winter and pasture that approaches 100% KY31 fescue during the summer. There has been no supplemental pasture or other forage fed to these animals. The clinical signs in the Tennant herd that highly suggest endophyte toxicity (fescue mycotoxicosis) include: swelling and pain at the coronary band (coronitis) and above, with cattle that stand in the creek during hot weather (Figures 5-8); poor shedding of winter coats (Figure 4); . patchy tail alopecia with loss of tail switch; birth of undersized calves; poor conception and calving rates: numerous vague diseases suggestive of immune dysfunction; 000023 20 EID151701 R G S000904 Tennant Farm Herd Health Investigation Cattle Team Report Other clinical signs that are consistent with endophyte toxicity include: fat necrosis, presumptive, as diagnosed by rectal palpation in cow #15; concomitant presence of copper deficiency; panting in cow #45, suggestive of hyperthermia; general failure of the cattle to thrive. Vasoconstriction is seen as a result of ingestion of a number of ergot-like alkaloids. The endophyte fungus associated with fescue mycotoxicosis is Acremonium coenophialum which produces perloline, perlolidine, N-acetyl loline, and N-formyl loline (Stuedemann, et al., 1985). These alkaloids accumulate in the fescue grass seed during growth, acquiring the highest concentrations at the greatest maturity of the grass. Additionally, the alkaloids may be found in stored hay. The coronitis problem observed in the videotapes (Figures 5 and 6) was typical of that produced by endophyte toxicity in the "fescue foot" syndrome. Signs of fescue foot generally start with reduced weight gain, or loss of weight, rough hair coat, arched back, and soreness in one or both rear limbs. Hyperemia of the coronary band occurs between the dewclaws and hooves and is generally accompanied by some swelling (Hemken, et al., 1984). Arching of the back was reported by the owner in the videotapes as "hunched" and "humped" posture in some cattle. Peripheral vasoconstriction from these vasoactive alkaloids results in decreased ability to dissipate heat during hot weather. This phenomenon is known as "summer slump" (Osborne, et al., 1992). The panting and drooling by a red cow (#45) in videotape #2 was suggestive of the summer slump syndrome. T: e birth of undersized calves to dams ingesting a diet high in endophyte-infested fescue has also been documented (Bolt, et ah, 1986). This may be by a mechanism similar to the vasoconstriction caused by the ergot-like alkaloids. Vasoconstriction of the uterine blood vessels may result in decreased circulation to the growing fetus. The effect has been documented in mice that were exposed to the endophyte during gestation. Mouse pups from females fed the material during gestation were a lighter birth weight, were delayed in post-partum development, and grew slower than control mice (Varney, et ah. 1991) Evidence that endophyte toxicity in fescue grazing cattle may be responsible for the loss of the tail switch is largely anecdotal, from veterinary practitioners that have experience with similarly affected cattle. The most severe forms of endophyte-induced vasoconstriction result in dry gangrene of the tail and digits (Radostits, et ah, 1994; Hemken. et ah, 1984). Reproductive effects from the ingestion of endophyte-infested fescue include incrementally depressed conception rates in cattle grazing infected pasture (Paterson, et ah, 1995). In the Tennant herd, the grazing of pastures with 100% KY31 fescue could potentially play a major role in the unacceptable conception rate. A poor conception rate 000024 21 EID151702 R G S000905 Tennant Farm Herd Health Investigation Cattle Team Report could also be secondary to a lack of intensive breeding management at the Tennant farm, where breeding occurs year round. The advanced age of most animals in the breeding herd could also be expected to be a cause for depressed fertility and an extended calving interval. Likely, a combination of all three factors have contributed to suboptimal fertility. The poor growth of the nursing calves could be multifactorial as well. The advanced age of the cows or a limited genetic potential in the herd could lead to poor milk production and poor calf growth. Similarly, lack of proper nutrition or a lack of a balanced diet among the brood cows could depress calf growth. The ingestion of endophyte-infested fescue could lead to depressed milk production due to prolactin inhibition (Hurley, et al,, 1980; Paterson, et al., 1995). The mechanism is considered to be due to the dopaminergic effects of the endophyte ergopeptides on prolactin production in the bovine pituitary gland (Schultze, et al., 1999). Endophyte intoxication is also considered to have an immunosuppresive effect by an apparently indirect route. Although anecdotal evidence exists to suggest that fescue intoxication is responsible for depressed immune function, there appears to be no apparent lack of immune response to vaccinations of cattle fed diets high in fescue (Rice, et al.. 1997). Indirectly, however, there is a relationship between immune function, endophyte intoxication and copper status of cattle on a high fescue diet (Dennis, et al., 1998; Saker. el al.. 1998). Steers grazing endophyte-infested tall fescue have lower serum copper concentrations than those not on a high-endophyte tall fescue diet, resulting in depressed monocyte-macrophage functions. This is apparently an effect of the decreased uptake of copper by the infected tall fescue plants. Another source has recorded depressed globulin levels in animals grazing infected pastures (Schultze, et al., 1999). Fat necrosis is a herd problem of cattle grazing tall fescue with high endophyte levels. The necrotic fat accumulations are mostly within the omental and retroperitoneal fat depots, and consist of hard masses of partly mineralized fat. Cow #15 had palpable masses within the abdominal cavity that were identified during the herd exam on April 7. 1999. These masses were most suggestive of masses of necrotic fat. The biochemical reaction causing the fat necrosis is unclear, however, in affected animals that are on a diet high in N-formyl and N-acetyl loline, there is a consistent reduction of circulating levels of cholesterol iStuedemann. et al., 1985). Animals with increased dietary levels of endophyte-infected fescue had decreased body condition and increased fat necrosis. Pinkeye Faceflies were a serious problem in the cattle in the videotaped recordings taken on the Tennant farm (Figures 1 and 2). Additionally, they have also been a problem in other years, according to the history provided by Mr. Tennant during the April 8, 1999 interview. Faceflies were seen in small numbers at the eyes of the cattle during the April 7, 1999 herd visit, representing an early spring infestation. The facefly, Musca autumnalis, is a vector for Moraxella bovis, the etiology of most bovine pinkeye. Control 000025 22 EID151703 R G S000906 Tennant Farm Herd Health Investigation Cattle Team Report of these insects is based on preventing the flies from feeding on the lacrimal secretions of the cattle. Insecticides and insecticidal ear tags, as applied during the visit, are effective controls. Secondarily, the insecticides tend to limit the spread of M. bovis between susceptible cattle during outbreaks of pinkeye. Severe facefly infestations can result in poor feed consumption and conversion among affected cattle. Some of the poor weight gain described in the history of the herd and the videos may be due to "fly worry" The manic stampeding of the cattle, that was described in one of the videotapes, was likely due to the large number of flies. Malnutrition Malnutrition was considered to be a significant problem in the Tennant herd. The owner had identified poor growth rates and infertility as specific problems among his cattle. This was considered a manifestation of protein-energy insufficiency and was reflected in the low body condition scores. Dietary requirements of beef cattle change dramatically during the life cycle of the animals and during the year, particularly with respect to such environmental and physiological stressors as fluctuating ambient temperatures and moisture conditions, lactation, growth, and breeding (Rice, 1991). The clinical impression of malnutrition that followed the visits of April 7 and 8 were confirmed by the results of the feed analysis of the hay and grain mix rations taken during the visits. Copper deficiency As indicated by the heavy metal analysis performed by two laboratories, on tissues from three animals, copper deficiency is a problem in the Tennant herd (Appendix B). In 1997, copper deficiency was diagnosed in a 4-year old Holstein cow and a 9-year old Holstein cow by the Animal Health Diagnostic Laboratory of Michigan State University in East Lansing, Michigan. In 1999, copper deficiency was revealed in tissues from a sacrificed 7-vear old red cow by the Laboratory of Large Animal Pathology and Toxicology of the University of Pennsylvania, Kennett Square, Pennsylvania. Furthermore, serum samples taken from 41 adult animals at the farm on April 7. 1999 revealed that 26 of the 41 (63%) had serum copper levels below that considered deficient at the reference laboratory (Table 4). Clinical signs of copper deficiency seen on the Tennant farm were lightening of the hair coats, poor quality hair on the cows, and overgrown hooves. This was reported by Mr. Tennant and shown in videotapes made of the herd. The clinical effects of copper deficiency are multiple, and the clinical and physiological effects of such deficiency are well documented (NRC, 1996). The effects include immune deficiencies, poor quality hair coats, increased fragility of bone and sudden death in extreme cases. During disease, copper deficiency can result in depressed levels of tumor necrosis factor (a cytokine) with resulting abnormal temperature responses to infections and depressed feed intake (Gengelbach, et al., 1997). 000026 23 EID151704 R G S000907 Tennant Farm Herd Health Investigation Cattle Team Report Immune deficiencies related to copper deficiency are multiple and include depressed acute phase protein response and lymphocyte responsiveness to mitogen stimulation (Arthington, et al.. 1996; Gengelbach. et al., 1997). As there has been no documentation of any copper supplementation of the herd, the deficiency is expected to be present at this time and to continue. The relationship between high endophyte fescue grazing and copper status in the feed has been described (Dennis, et al., 1998). Copper deficiency is widespread throughout the United States (Dargatz, et al., 1999). Toxicology issues Based upon the draft report entitled Dry Run Creek, 1997 from the Environmental Response Team of the US EPA, carnivorous, piscivorous, omnivorous, insectivorous and herbivorous mammals in the Dry Run Creek study area are at increased health risk due to exposure to metals, fluoride and trichlorofluoromethane. At least with regard to metals of concern, diagnostic evaluation of tissue and fluids collected from three animals from the Tennant farm (two submitted to Michigan State University and one submitted to New Bolton Center) do not suggest elevated concentrations of any metal. In addition, urine (two samples submitted to Michigan State University) and bone (one sample submitted to Michigan State University from the cow evaluated at New Bolton Center) samples for fluoride analysis did not measure concentrations above expected or "background'' values. The urinary fluoride values were 6.60 ppm and 5.55 ppm and one bone fluoride value was 1090 ppm (expressed on a fat free, dry weight basis). Urine and bone fluoride concentrations in adult cattle consistent with fluorosis are 15 to 20 ppm or greater and 3000 ppm or greater (expressed on a fat free, dry weight basis), respectively (Osweiler et al., 1985). In addition, there was no clinical evidence of chronic fluorosis in the Tennant herd. Exposure to trichlorofluoromethane was considered to be a default risk factor based upon a lack of toxicologic benchmarks for this compound. However, available toxicity data derived from inhalation studies using common laboratory animals indicate that chlorofluorocarbons such as trichlorofluoromethane have low acute and chronic toxicity (Magda. 1999). Signs associated with acute toxicity are reversible effects on the central nervous system such as lethargy and incoordination. Chlorofluorocarbons are not developmental toxicants, do not affect reproductive performance and are not genotoxic. 6.0 RECOMMENDATIONS The most important general recommendation from the cattle team, after completion of the herd visit and review of the investigatory data, would be for the owner of the Tennant herd to engage veterinary and nutritional consultants in the design of a herd health program. 000027 24 EID151705 R G S000908 Tennant Farm Herd Health Investigation Cattle Team Report Endophyte toxicity. With dietary forage that contains nearly 100% KY31 fescue, fed during all seasons, the immediate goal would be for the dilution of the endophyte in the diet by mixing the hay and pasture with either non-endophyte infested fescue or another different forage source. A short-term goal would be to reduce the content of endophyteinfested hay and pasture to 50% or less of the diet, primarily by supplementing the diet with higher quality forage. An acceptable long-term plan would be to replace the fescue pasture and hay fields with a non-endophyte-infested forage crop or crops over time, again with the goal of diluting the dietary endophyte consumption. Crop management should be attempted under the supervision of specialists in forage and grazing management. Pinkeye: Prevention of pinkeye (keratoconjunctivitis) in the immediate future should be attempted by an approach that limits the spread of the infectious organism, Moraxella bovis, from carrier cattle to susceptible cattle. Effective fly control, through the use of insecticide-impregnated eartags or some other proven method, is perhaps the most effective control method that should be practiced during the warm months. The insecticide-laden eartags should be rotated every year in order to circumvent the development of insecticide resistance by the local facefly population. An additional long-term approach that may be employed is the breeding for cattle that have dark faces. By selecting for dark-faced cattle, the corneal damage that is caused by the ultraviolet light of the sun is minimized. However, while minimizing the damage to the corneas of the cattle may prevent overt lesions of keratoconjunctivitis, the loss of condition from severe fly infestations will not be avoided by selecting for face color in cattle. Thus regardless of the type of cattle in the herd, insecticide fly repellants should be used every year. Malnutrition: Rations and pasture nutrition should be planned with the assistance of a specialist in beef cattle nutrition. In the Tennant herd, there was evidence of gross protein-energy malnutrition, as well as concerns about macromineral (calcium, phosphorus, and magnesium) nutrition. Trace mineral and vitamin deficiencies are common to many beef herds and should also be addressed. In the development of a ration for this herd, particular attention should be paid by the nutritionist to the local trace mineral and macromineral deficiencies. Copper deficiency: Copper is a trace mineral that is crucial to the health of cattle yet is missing from the diet of cattle in many areas of the North America. As copper deficiency was suspected clinically and confirmed biochemically in the Tennant herd, the provision of a high copper trace mineral supplement, again under the supervision of a beef cattle nutritionist, should be a priority year round. This is especially critical considering the prevalence of endophyte-infested fescue on the Tennant farm. 000026 25 EID151706 R G S000909 Tennant Farm Herd Health Investigation Cattle Team Report 7.0 CONCLUSION There was conclusive evidence that the Tennant cattle herd was, and continues to be, suffering from four major disease entities, some of which were potentially interrelated: endophyte toxicity (fescue mycotoxicosis), pinkeye, malnutrition, and copper deficiency. As substantiated by the clinical and laboratory findings, and historical data, these four conditions readily account for the chronic herd health problems on the Tennant farm. The herd health investigation revealed deficiencies in herd management, including poor nutrition, inadequate veterinary care, and lack of fly control. The lack of vaccination and internal parasite control programs did not appear to have a substantial impact on this relatively isolated herd. Despite an exhaustive review of historical and contemporary herd data, there was no evidence of toxicity associated with chemical contamination of the environment. 8.0 REFERENCES Endophyte Toxicity References 1. Bolt DJ, Bond J: Effects in pregnant beef heifers grazing fungus-infected tall fescue on calf birth weight, milk yield, and calf growth. J Anim Sei 63(suppl):297, 1986. 2. Dennis SB, Allen VG, Saker KE. Fontenot JP, Ayad JYM, Brown CP: Influence of Neotxphodiuni coeiiophiahtni_on copper concentration in tall fescue. J Anim Sei 76:2687-2693, 1998. 3. Hemken RW, Jackson JA, Boling JA: Toxic factors in tall fescue. J Anim Sei 58:1011-1016,1984. 4. Hurley WL. Convey EM. Leung K, Hemken RW: Bovine prolactin. TSH, T4 and T3 concentrations as affected by tall fescue summer toxicosis and temperature. J Anim Sci 51:374-379. 1980. 5. Osborne TG, Schmidt SP, Marple DN, Rahe CH, Steenstra JR: Effect of consuming fungus-infected and fungus-free fescue and ergotaminc tartrate on selected physiological variables of cattle in environmentally controlled conditions. J Anim Sci 70:2501-2509, 1992. 6. Paterson J, Forcherio C, Larson B, Samford M, Kerley M: The effects of fescue toxicosis on beef cattle productivity. J Anim Sci 73:889-898, 1995. 7. Radostits OM, Blood DC. Gay CC: Diseases caused by toxins in plants, fungi, cyanobacteria, clavibacteria, insects, and animals. In: Veterinary Medicine: A 000029 26 EID151707 RGS000910 Tennant Farm Herd Health Investigation Cattle Team Report textbook of the diseases of cattle, sheep, pigs, goats, and horses, 8lh ed., Radostits OM, Blood DC, Gay CC, editors. Baillire Tindall, Philadelphia 1532-1611, 1994. 8. Rice RL, Blodgett DJ, Schurig GG, Swecker WS, Fontenot JP, Allen VG, Akers RM: Evaluation of humoral immune responses in cattle grazing endophyte-infected or endophyte-free fescue. Vet Immun Immunopathol 59:285-291, 1997. 9. Saker KE. Allen VG, Kalnitsky J, Thatcher CD, Swecker, Jr. WS, Fontenot, JP: Monocyte immune cell response and copper status in beef steers that grazed endophyte-infected tall fescue. J Anim Sci 76:2694-2700, 1998. 10. Schultze AE, Rohrbach BW, Fribourg HA, Waller JC, Oliver JW: Alterations in bovine serum biochemistry profiles associated with prolonged consumption of endophyte-infected tall fescue. Vet Human Toxicol 41:133-139, 1999. 11. Stuedemann JA, Rumsey TS, Bond J, Wilkinson SR, Bush LP, Williams DJ, Caudle AB: Association of blood cholesterol with occurrence of fat necrosis in cows and tall fescue summer toxicosis in steers. Am J Vet Res 46:1990-1995, 1985. 12. Varney DR, Varney LA, Zavos PM. Wiglesworth MD, Siegel MR: Tall fescue endophyte: effect on congenital development and pup growth in mice. J Dairy Sci 74:460-466,1991. Malnutrition References 13. Rice LE: The effects of nutrition on reproductive performance in beef cattle. Vet Clin North Amer - Food Anim Pract 7:1-26. 1991. Copper Deficiency References 14. Arthington JD. Corah LR, Blecha F: The effect of molybdenum-induced copper deficiency on acute-phase protein concentrations, superoxide dismutase activity, leukocyte numbers, and lymphocyte proliferation in beef heifers inoculated with bovine herpesvirus-1. J Anim Sci 74:211-217, 1996. 15. Dargatz DA, Garry FB, Clark GB: Serum copper concentrations in beef cows and heifers. JAVMA 215:1828-1832, 1999. 16. Dennis SB, Allen VG, Saker KE, Fontenot JP, Ayad JYM, Brown CP: Influence of Neoth\phodium coenophialum_on copper concentration in tall fescue. J Anim Sci 76:2687-2693, 1998. 000030 27 EID151708 R G S000911 Tennant Farm Herd Health Investigation Cattle Team Report 17. Gengelbach GP, Ward JD, Spears JW, Brown TT: Effects of copper deficiency and copper deficiency coupled with high dietary iron or molybdenum on phagocytic cell function and response of calves to a respiratory disease challenge. J Anim Sci 75:1112-1118, 1997. 18. National Research Council: Minerals, in: Nutrient Requirements of Beef. 7th ed. National Academy Press, Washington, D. C. 54-74, 1996. Toxicology Issues References 19. Magda S: Fluorocarbons. In: Toxicology, 1st ed., Marquardt H, Schafer SG, McClellan R and Welsch F, editors., Academic Press, San Diego, 659-662, 1999. 20. Osweiler GD, Carson TL, Buck WB, Van Gelder GA, editors: Fluoride. Clinical and Diagnostic Toxicology. Kendall/Hunt Publishing, Dubuque, IA, 183-188, 1985. 21. U.S.E.P.A. Environmental Response Team: Dry Run Creek. 1997 (draft). 000031 28 EID151709 R G S000912 Tennant Farm Herd Health Investigation Cattle Team Report 9.0 GRAPHS Graph 1: Individual Cattle Plasma Prolactin (April 7, 1999). Heparinized blood samples taken from 41 adult cattle during the cattle team's visit to the Tennant farm were analyzed for the endophyteresponsive hormone, prolactin. Graph illustrates the laboratory's reference means for endophyte-free (171.6 ng/mL) and endophyteinfested (105.8 ng/mL) April pasture. The average plasma prolactin value for the Tennant herd (110.1 ng/mL) was similar to the endophyte-infested reference. These results were highly supportive of the diagnosis of endophyte mycotoxicosis in the Tennant herd. (Testing laboratory: Dr. Neil Schrick, University of Tennessee) 000032 29 EID151710 R G S000913 Graph 1: Individual Cattle Plasma Prolactin (April 7, 1999) 600 A 550 500 450 400 I 350 "&> . 300 +<U+0 S 250 Cl 200 A A A 150 100 50 0 A A A AAAA AA A A A A A *A * AA T-- P-- T 1------- 1------- r ' < ,m~r 5 I 1------- , --- 1 1 1-- 10 15 20 25 Animal Number A A A A A -- I-------- 1-----------------1--------1--------j r i 30 35 i--------1------- i------- 1------- 1- r 40 k Prolactin] 171.6 110.1 105.8 45 Tennant Farm Herd Health Investigation Cattle Team Report 10.0 FIGURES Figure 1: Faceflies on the head of a calf. Facefly (Musca autumnalis) infestation was considered to be a major disease problem on the Tennant farm. In addition to the constant harassment by flies, this calf had severe bilateral keratoconjunctivitis and appeared to be clinically blind on the videotape (photo was taken from Tennant videotape # 2 ). Figure 2: Faceflies on the head of a cow. This cow was the dam of the calf in figure 1. Like her calf, she had severe bilateral keratoconjunctivitis, (photo was taken from Tennant videotape #2). Figure 3: Left eye of a calf with faceflies and central corneal opacity. Faceflies are a vector for the bacterial agents of pinkeye (keratoconjunctivitis) such as Moraxella bovis. Central corneal opacities may progress to diffuse corneal ulceration and clinical blindness, (photo was taken from Tennant videotape #2). Figure 4: Delayed shedding of the winter coat on a beef cow. The videotapes presented multiple cattle with delayed shedding of winter coats. This is a clinical sign often associated with endophyte toxicity as well as nutritional deficiencies, (photo was taken from Tennant videotape #2). Figure 5: Alopecia and erythema above the coronary band (coronitis). This lesion was present in several cattle in the videotapes. It is a common sign of endophyte toxicity and is generally referred to as "fescue foot", (photo was taken from Tennant videotape #2). Figure 6: Alopecia and erythema above the coronary band (coronitis). This example of "fescue foot" is similar to that presented in figure 5. (photo was taken from Tennant videotape #2). Figure 7: Three cattle standing in the creek. A unusual predilection for standing in water was described for the Tennant cattle in the summer. This behavior is known to be associated with endophyte toxicity as it provides some relief for both coronitis ("fescue foot") and hyperthermia ("summer fescue"), (photo was taken from Tennant videotape #2). Figure 8: Two cattle standing in a puddle. As described for figure 7. standing in water provides some clinical relief to cattle with coronitis. Although non-specific, a predilection for this behavior is associated with "fescue foot", (photo was taken from Tennant videotape # 2). 31 000034 EID151712 R G S000915 3f *f * m # 3* t }i /'<j Tennant Farm Herd Health Investigation Cattle Team Report 11.0 TABLES Table 1: Review of Tennant Farm Videotapes Table 2: Individual Animal Data: Clinical Signs Table 3: Individual Animal Data: Hematology (erythron, platelets) Table 4: Individual Animal Data: Hematology (leukon), and Special Chemistry Table 5: Individual Animal Data: Clinical Chemistry Table 6: Individual Animal Data: Serology and Fecal Exam Table 7: Tennant Farm Grain and Hay: Nutritional Analysis 000039 EID151717 R G S000920 Tennant Farm Herd Health Investigation Cattle Team Report Table 1. Review of Tennant Farm Videotapes Tape #1 (cases 1 -18): Tennant Farm: New England, Wood Co., WVa - January & February, 1997__________________ # Subject Animal(s) Presentation Diagnosis/Comment 1 Cow, Hereford neck: alopecia, scaling, probable lice, described as "humped- "humped-up" difficult to appreciate up". . (snowing) on tape. 2 Cow, Hereford corneal opacity, comeal scar, probably secondary to (snowing) pinkeye. 3 2 black bulls possible discoloration not clear from tape. of hair (not clear), (snowing) 4 Cow, Hereford thin, very thin, probably due to described as poor teeth, decreased food consumption and (snowing) mastication problem, although teeth not seen on video, age unknown; cause of possible teeth problem unknown. 5 Cow, Hereford possible hunched back not clear from tape. (not clear), (snowing) 6 Cow (tan) alopecia, tail switch, tail hair loss differential diagnosis (snowing) would include mechanical, lice, fescue toxicosis, and selenium toxicosis. 7 Cow . alopecia, tail switch, tail hair loss differential diagnosis (snowing) would include mechanical, lice, fescue toxicosis, and selenium toxicosis. 8 Calf, black dead (dated 2/9/97) in hooves a little longer than normal, snow, but not unusual, overgrown hooves, "cold cataracts" (post mortem), bilateral lens opacities, teeth normal, black/brown teeth, fecal mucous normal for stagnant mucous on feces, feces in rectum, necropsy: no other lesions, . stated: had diarrhea necropsy: lack of fat; serous atrophy of fat: emaciation, probably due to starvation. Table 1: Review of Tennant Farm Videotapes 000040 7 EID151718 R G S000921 Tennant Farm Herd Health Investigation Cattle Team Report Table 1 (continued): Review of Tennant Farm Videotapes Tape #1 (cases 1- 18): Tennant Farm: New England, Wood Co., WVa - January & February, 1997__________________ # Subject Animal(s) 9 Calf, black/white face Presentation neck: alopecia, diarrhea, lens or corneal opacities Diagnosis/Comment probable lice, cause of diarrhea unknown, probable comeal scar (hard to tell). 10 Cow, Hereford thin, comeal opacity probably secondary comeal opacity, to pinkeye, above hooves: alopecia differential diagnosis of hoof and hyperemia, lesions include mechanical diarrhea dermatitis, fescue toxicosis, and moist dermatitis due to unknown cause of diarrhea and thinning unknown, 11 Cow, black neck: alopecia probable lice. 12 Cow, red neck: alopecia probable lice. 13 Cow , red 14 Cow, red 15 Cow, red slobbering, losing cud (on ground), urinating, tail switch alopecia neck: alopecia, tail switch: short hair neck: alopecia slobbering/urinating differential diagnosis includes hardware and cholinesterase inhibition (less likely due to individual cow affected), alopecia differential diagnosis includes mechanical, lice, and fescue or selenium toxicosis. neck: probable lice, tail hair differential diagnosis includes mechanical, lice, and fescue or selenium toxicosis. probable lice. 16 Cow , red odd chewing behavior not clear from tape. 17 Cow, Hereford possible hunched back not clear from tape; differential diagnosis includes hardware disease (traumatic reticulopericarditis). Table I: Review of Tennant Farm Videotapes 000041 36 EID151719 R G S000922 Tennant Farm Herd Health Investigation Cattle Team Report Table 1 (continued): Review of Tennant Farm Videotapes Tape #1 (cases 1 - 18): Tennant Farm: New England, Wood Co., WVa - January & February, 1997__________________ # Subject Animal(s) Presentation Diagnosis/Comment 18 Cow, red dead in bam, (is this the 9-year-old cow (same as #13) corneal opacity, submitted to Michigan?), necropsy: no other comeal opacity may be postmortem lesions artifact, . necropsy: serous atrophy of fat, noted: normal postmortem changes include moderate to severe autolysis, pseudomelanosis of intestines, interlobular emphysema (agonal), . also noted: normal front teeth large gallbladder suggests period of anorexia, cause of death unknown. Table I: Review o f Tennant Farm Videotapes 000042 39 EID151720 RGS000923 Tennant Farm Herd Health Investigation Cattle Team Report Table 1 (continued): Review of Tennant Farm Videotapes Tape #2 (cases 19- 60): Dry Run Harris PC. Wood Co. - Off North Fork of ____________ Lee Creek, New England, WVA_________________ # Subject Presentation Diagnosis/Comments Animal(s) 19 Calves, multiple comeal opacities of fly problem with secondary varying severity; some pinkeye, with exudate and/or hair coat shedding and coloring blepharospasm, fly problem (summer), probably due to a nutritional problem. some with poorly shedded and/or light- colored coats 20 Fish (sucker?) dead incidental death: no diagnosis possible. 21 Small mammal skeleton incidental death: no diagnosis (raccoon?) possible. 22 Cow comeal opacity comeal scar secondary to pinkeye. 23 Calf 24 Calf 25 Calf 26 Snake 27 Calf 28 Fish 29 Calf 30 Calf comeal opacities, possible head tilt (not clear) corneal opacities, . flies dead (-1 month-old), corneal opacity, (burning carcass) dead corneal opacities, fly problem dead corneal opacity corneal opacity comeal scar secondary to pinkeye, . head tilt not clear on tape. comeal scar secondary to pinkeye. . cause of death unknown, comeal scar secondary to pinkeye. incidental death: no diagnosis possible. . fly problem with secondary pinkeye. incidental death: no diagnosis possible. comeal scar secondary to pinkeye. corneal scar secondary to pinkeye. Table 1: Review o f Tennant Farm Videotapes 000043 0 EID151721 R G S000924 Tennant Farm Herd Health Investigation Cattle Team Report Table 1 (continued): Review of Tennant Farm Videotapes Tape #2 (cases 19 - 60): Dry Run Harris PC. Wood Co. - Off North Fork of _________________ Lee Creek, New England, WYA_________________ # Subject Presentation Diagnosis/Comments Animal(s) 31 Cow dead, worn incisors, . diarrhea, cause of diarrhea, emaciation and death unknown, worn incisors may be age or feed emaciated, sunken eyes related, (dehydrated?), serous lung emphysema was agonal and atrophy of fat (heart), insignificant. . agonal lung emphysema 32 Crow dead incidental death: no diagnosis possible. 33 Crow dead incidental death: no diagnosis possible. 34 Calves, corneal opacity comeal scar secondary to pinkeye. multiple 35 Cow corneal opacity, comeal scar secondary to pinkeye, thin diagnosis of the cause of thin condition not possible. 36 Fish dying incidental death: no diagnosis possible. 37 Crayfish . dead incidental death: no diagnosis possible. 38 Salamander . dead incidental death: no diagnosis possible. 39 Toad dead incidental death: no diagnosis possible. 40 Newborn calf contracted tendons, big hocks difficult to evaluate from tape; may be congenital contracted tendons. 41 Deer dead . incidental death: no diagnosis possible. 42 Cow dead (rotten) no diagnosis possible. 43 Deer 44 Deer . rotten carcass bones incidental death: no diagnosis possible. incidental death: no diagnosis possible. Table 1: Review o f Tennant Farm Videotapes 000044 n EID151722 R G S000925 Tennant Farm Herd Health Investigation Cattle Team Report Table l (continued): Review of Tennant Farm Videotapes Tape #2 (cases 19 - 60): Dry Run Harris PG. Wood Co. - Off North Fork of ____________ Lee Creek, New England, WYA_________________ # Subject Presentation Diagnosis/Comments Animal(s) 45 Cow panting; increased respiration differental diagnosis includes hyperthermia due to summer fescue toxicosis, 46 Cow diarrhea diagnosis uncertain. cause of diarrhea unknown. 47 Raccoon 48 Deer 49 Deer 50 Rabbit 51 Turkey 52 Hawk 53 Fish 54 Cow skull skeleton dead (fresh), bloody nose dead dead dead dead lumpy udder incidental death: no diagnosis possible. incidental death: no diagnosis possible. death possibly due to epizootic hemorrhagic disease of deer (EHD). incidental death: no diagnosis possible. incidental death: no diagnosis possible. incidental death: no diagnosis possible. incidental death: no diagnosis possible. unclear from videotape. 55 Cows, multiple above hooves: alopecia, differential diagnosis includes possibly erythema, mechanical dermatitis, fescue overgrown hooves toxicosis, and moist dermatitis due to unknown, hoof length not significant. 56 Bull, cows, corneal opacities, comeal scar secondary to pinkeye, multiple alopecia and erythema hoof differential diagnosis includes above hooves mechanical dermatitis, fescue toxicosis, and moist dermatitis due to unknown. 57 Cow lumpy udder unclear from videotape. Table I: Review o f Tennant Farm Videotapes 000045 EID151723 R G S000926 Tennant Farm Herd Health Investigation Cattle Team Repon Table 1 (continued): Review of Tennant Farm Videotapes Tape #2 (cases 19 - 60): Dry Run Harris PC. Wood Co. - Off North Fork of _________________ Lee Creek, New England, WVA_________________ # Subject Presentation Diagnosis/Comments Animal (s) 58 Cows, multiple above hooves: alopecia, hoof differential diagnosis includes possibly erythema, mechanical dermatitis, fescue . poor shedding of coat toxicosis, and moist dermatitis due in some animals to unknown, poor hair coat shedding suggests nutritional problem or fescue mycotoxicosis. 59 Cow (tan) unusual gait lame due to unknown cause. 60 Cows, heifers, thin. multiple delayed shedding definitive diagnosis not possible from tape - probably a nutritional deficiency. Note: This table excludes many of the clinical signs and pathological conditions that were proposed by the videotapes' narrator but were considered by the cattle team to be inaccurate. For example, the team did not agree with the narrator's assertion that blackened teeth or patchy melanosis were abnormal. Also numerous speculative comments regarding normal organs, during the dissections shown in the videotapes, were not included in this table. Table I: Review o f Tennant Farm Videotapes ~00004l> EID151724 R G S000927 Tennant Farm Herd Health Investigation Cattle Team Report ooo o W Q CO Individual Animal Data: Cattle from Tennant Farm (Washington, West Virginia) Table 2: Clinical Signs Animal Age Preg. Number Status Description Body Condition Score (1 - 1(1) Clinical Signs years I Girth BCS (in .) (1-10) I >9 4 m yellow - 3 possible lice 2 >9 open red, white face - 3 lice, corneal scar (r) 3 > 9 open yellow, white face - 3 (large uterus) 4 >9 7 m red, white face 70 4 lice 5 >9 6 m red, white face 75 4 corneal scar (1) 6 >9 3 m red, white face - 4- 7 >9 6 m red, white face 71 4 hair loss (switch) 8 > 9 open red, white face -4 9 > 9 open red, white face 70 4 (muzzle: melanin considered normal) 10 >9 term red, white face 70 3 - 11 >9 8.5 m red, white face - 3- 12 > 9 term red, white face 72 3 light coat color; small corneal scar (r) 13 >9 open red, white face 66 2 slight corneal scar (1), 14 >9 calf red, white face 78 4 alopecia (brisket) 15 >9 open yellow 71 2 palpation difficult (fat necrosis?) 16 6 open black 68 3 - 17 >9 18 >9 4 m red, white face 4 m red, white face ....'. 7_.8.. . 77 mammary gland lumps (right front)(abscesses?) 19 >9 J Calf red, brockleface 68 3 slight corneal scars (1 and r) 20 >9 6 m red, white face 21 >9 __5_m__ red, white face ....7 4 ...... .... .3. ..... 69 small corneal scar (l)(5 mm) 22 >9 5 m red, brockleface 76 4 mass (1 2 cm) prescap. neck (r) - lanced: brown serum + hair 23 - bull black - 4- 24 >9 6 m red, white face 78 4 _ hair loss (neck) t Photo. # ______ 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18,53 19 20 21 22, 49-51 23 24 Data Tables EID151725 Tennant Farm Herd Health Investigation Cattle Team Report Individual Animal Data: Cattle from Tennant Farm (Washington, West Virginia) Table 2: Clinical Sign.v(continued) Animal Ago Prog. Description Number Status Hotly Condition Score (1 10) Clinical Signs Photo # years | Girth BCS (in.) (1-10) 25 4 5 m black ' 7 2 .... 4 corneal scar (1) 25,44 26 6 hull black - 5 corneal opacity (1) 26, 45 27 >9 8 m red, brockleface 64 3- 27 28 7 8 m black, white face 74 4- 28 29 >9 term red, brockleface 77 2 mammary gland lumps (left rear) (abscesses?) 29 30 >9 8 m red, white face 72 3 lice, hair loss (abd.), extra teat, corneal opacity (l=3mm; r=5mm) 30, 46 31 8 6 m red, white face 77 5 corneal scar (r) (2 mm); (tongue: melanin considered normal) 31,52 32 >9 6 in black, white face 70 3 lice 32 33 >9 34 >9 8 m red, white face 8 m red, brockleface 72 73 3T- 33 34 35 >9 5 m red, white face 72 3 - 35 36 >9 8 m red, white face 75 3 corneal scar (1), severe 36 37 7 calf red, brockleface 77 4 corneal scar (r) (1 cm) 37,47 38 >9 7 in red, white face 73 4 - 38 39 >9 calf red, white face 68 2 - 39 40 4 6 m black 75 4 light hair color, corneal opacity (1) (2mm) 40,48 41 4 steer black, white face 71 - lame (Ir), corneal opacity (r) severe; phthysical eye (1) 41 1 cow - red, whiteface - - cow escaped run before being tagged or examined calves < 6 m - 12 mise, calves - - 5 bigger calves not tagged; 7 smaller calves tagged 42 - Cattle examinations conducted on April 7, 1999. Age determined by teeth examination (Dr. Moisan); Pregnancy Status determined by rectal palpation (Dr. Munson): Girth measured by tape (Dr. Habecker); BCS assigned by Drs. Moisan and Munson; Clinical Signs identified by cattle team (Drs. Davis-Heiler, Habecker, Moisan, Munson, Poppenga, Sykes); Photos taken by Dr. Sykes. Cow #22: Gross diagnosis (no histology) of skin lesion: epidermal inclusion cyst (incidental lesion of no significance). O CO Data Tables EID151726 Tennani Farm Herd Health Investigation Cattle Team Report Individual Animal Data: Cattle from Tennant Farm (Washington, West Virginia) Table J: Hematology (Erythron, Platelets) Animal Age Preg. It 111' Hgb Hit MCV MCH M t'HC RDU 1'I.T MPV PCT l'l)\V Number Status years I xlOVul ..t/ 1 % fl Pg g/dl % xlOVul n % Reference 5-10 8-15 24- 40-60 Range: 46 1 4 4 m 5.85 9.91 28.8 49.3 16.9 34.4 18.3 378 7.04 .266 15.9 2 3 open 4.68 8.82 26.0 55.6 18.8 33.8 19.5 186 9.10 .170 19.0 3 >9 open 6.22 12.0 34.8 55.9 19.3 34.6 17.8 233 8.50 .198 18.9 4 >9 7m 5.81 10.4 29.7 51.1 17.9 35.1 19.3 259 8.08 .209 18.4 5 >9 6 m 5.86 9.87 29.2 49.8 16.9 33.8 18.6 142 8.82 .126 19.7 6 >9 3 m 5.21 10.1 28.8 55.2 19.4 35.1 19.3 201 8.98 .180 18.7 7 >9 6m 5.06 10.6 29.3 58.0 21.0 36.1 17.8 378 6.49 .246 16.9 8 >9 open 4.44 9.58 26.6 59.9 21.6 36.0 19.2 448 6.42 .287 17.2 9 >9 open 4.48 8.66 25.2 56.3 19.3 34.3 20.2 184 13.1 .240 23.4 10 >9 term 5.23 10.5 29.7 56.9 20.1 35.3 17.1 369 8.02 .296 18.1 11 >9 8.5 m 4.81 9.84 28.2 58.6 20.4 34.9 17.8 142 - - - 12 >9 term 4.99 9.41 27.9 55.9 18.9 33.7 19.0 113 10.9 .123 21.8 13 >9 open 4.29 8.62 23.7 55.2 20.1 36.4 20.4 595 6.73 .400 17.7 14 >9 calf 6.04 12.0 33.7 55.8 19.9 35.7 18.8 361 8.91 .321 19.2 15 >9 pregn. 4.65 8.41 24.9 53.6 18.1 33.7 20.6 238 7.73 .184 19.2 16 6 open 5.47 9.58 28.2 ; 51.5 17.5 34.0 20.0 284 8.27 .235 18.3 17 >9 4 m 6.16 3.1 " y i n " ] 61.1 ...21 2 34/7 19.2.... 87.4 - - - 18 >9 4 m 6.30 12.5 35.2 55.9 19.9 35.6 19.4 368 7.71 .283 17.2 19 >9 calf 4.92 9.98 29.1 59.2 20.3 34.3 ' 17.3 317 5.75 .182 16.1 20 >9 6m 4.85 9.51 28.0 57.7 19.6 34.0 18.0 275 7.55 .207 18.6 21 >9 5 m 5.80 10.7 30.9 53.2 18.5 "'34.8 19.0 320 8.30 .266 19.4 22 >9 5 m 5.55 0,5 ""t 30.3_ ..*54.7 18.9 34.6 .....9.1 391 7.24 .283 18.2 i 23 bull 6.13 \ 2 1*34.8 " 56.7 20.3 35.7 j 19.0 i 283 8.74 .248 20.9 24 >9 6m 6.02 [ 11.8 33.5 55.7 19.6 35.2 21.8 .....270 8.23 .222 17.9 Data Tables EID151728 ! ~ --------- ---- - Tennant Farm Herd Health Investigation i Cattle Team Report Individual Animal Data: Catje from Tennant Farm (Washington, West Virginia) Table J: Hematology (Erythwn, Timelets) (continued) Animal Age Prejj. ICIIC Hub Hit MCV MCH MCHC RDU p i / r MPV IT T PDW Number Status years I xltfVul 8/dl % n Pg g/dl % xl03/ul n % Reference 5-10 24- 40-60 Range: 46 25 4 5 m 6.93 12.5 35.9 51.8 18.0 34.8 18.8 231 8.33 .193 18.1 26 6 Ml 6.13 11.4 33.1 54.0 18.5 34.3 18.4 278 7.71 .215 18.2 27 >9 8 m 4.90 9.88 28.3 57.8 20.1 34.9 18.3 406 7.05 .286 17.8 28 7 8 m 6.12 11.7 34.1 55.7 19.2 34.4 17.8 180 7.18 .129 17.6 29 >9 term 5.37 9.93 28.7 i 53.4 18.5 34,6 19.2 220 9.57 j 311 24.4 30 >9 8 m 5.08 10.2 28.6 56.3 20.0 ~35.6 20.1 4 0 ] _ 7.09 .288 16.9 3I 8 6 m ' 6 03 ~ 12.6 36.0 ..."59.7.... ...'20.9 ....35.1..... ! 19.5 .... 2 7 8.07 .175 ; 18.2 32 >9 6 m 6.17 n .3 ... 32.4 i 52.5 i 18.2 34.7 19.2 138 7.33 .101 19.5 33 >9 8 m 5.42 10.1 30.1 55.5 i 18.7 33.6 17.8 186 7.77 .145 20.2 34 >9 8 m 5.41 11.6 33.0 60.9 21.4 35.1 18.2 361 7.20 .260 17.8 35 >9 5m 5.86 11.1 732.2 i~ ~5. 19.0 34.6_ 21.2 246 6.33 .156 16.8 36 >9 8 m 4~34~~ 9.44 "2673 j 6 0 6 ....... 2 I T .. "3 5 .9 18.1 _ ! 315 ; 8.00 .252 18.0 37 7 calf 5.48 T0.9 30.9 ~ 56.4 19.8 35.1 18.8 363 ! 8.56 .310 19.5 38 >9 7 m 5.88 11.5 32.2 54.8 196 35.7 18.3 397 j 5.82 .231 16.7 39 >9 calf 4.68 8.34 24.5 52.4 17.8 34.1 18.5 345 6.28 .217 16.4 oo 40 4 6 m 6.94 12.4 34.4 .. 49.6 .. .. 17.9 ... 36.1 _ 19.1 j 449 6.65 .298 \7 A 41 4 steer 6.78 12.2 34.7 18.0 ... 35.2 ' 20.0 298 6.94 .207 17.6 Blood collected by jugular venipuncture during examination while cattle restrained in a head-hold device. Blood refrigerated for approximately 36 hours before hematology was run on an Abbott Cell-Dyne 3500. Reference values are for cattle (New Bolton Center), but not from the Cell-Dyne 3500. Q i CO Data Tables I Tennant Farm Herd Health Investigation Cattle Team Report oooo Vt 0 CO Individual Animal Data: Cattle from Tennant Farm (Washington, West Virginia) Table 4: Hematology (leukon), Special Chemistry, ami Fecal Exam Animal Ago Prog. WIK Number Status Ncut Lymph \lGho Eos Iluso years | Reference Range: 14 4m 2 3 open 3 >9 open 4 >9 7 m 5 >9 6 m 6 >9 3 m 7 >9 6 m 8 >9 open 9 >9 open 10 >9 term tl >9 8.5 m 12 >9 term 13 >9 open 14 >9 calf 15 >9 pregn. 16 6 open 17 >9 4 m 18 >9 4 m 19 >9 calf 20 >9 6 m 21 >9 5 m 22 >9 5 m 23 - 24 >9 bull 6m xIO^ul 4-12 8.74 8.46 7.10 6.32 6.69 8.48 6.80 7.38 4.81 6.54 6.25 6.13 5.37 5.21 4.29 7.38 6.83 8.44 5.28 3.59 7.90 5.58 6.02 5.84 % %%% % 15-45 45-75 2-7 2-20 0-2 35.9 28.7 ~ 7.48 27.9 .016 32.0 44.2 9.36 14.4 .037 63.6 5.39 10.0 21.0 .024 42.5 28.9 7.39 21.1 .109 31.6 29.4 7.41 31.6 0.00 35.6 25.1 7.07 32.3 0.00 58.9 6.15 7.42 27.5 .029 53.7 16.3 .207 29.8 0.00 43.9 33.6 9.93 12.6 .054 62.2 15.3 10.5 11.8 .113 34.8 35.8 9.72 19.6 .061 45.7 33.0 7.75 13.5 .037 61.4 18.0 10.0 10.4 .151 31.2 41.9 8.25 18.5 .137 56.5 25.0 12.1 j 5.89 .428 49.7 33.7 1(17 1 5.76-"- ,..... .-2-4-0-- 44.8 27.6 10.2 45.1.... " T u ' ' 5.23 26.0 0.00 57.2 19.4 11.3 11.9 .058 53.4 19.2 8.19 I 19.2 0.00 32.0 22.6 .. 4 68 ] 40.7 0.00 44.6 14.3 ... 12.1 "28.9.... i.....167'" ' 59.1 '' 22 " 7.31 12.4 0.00 42.5 25.5 10.6 21.4 .028 Prolactin pldsnci ng/ml 171.6 (mean) 58.70 137.60 124.68 197.19 48.34 37.01 51.61 40.21 113.08 562.10 106.50 124.43 38.18 54.40 141.36 32.04 92.87 142.25 42.28 335 47.37 62.95 89.60 87.94 Cu scrum ppm 0.61.20 .282 .305 .518 .593 .693 .667 .473 .633 .518 .525 .344 .462 .586 .549 .244 .487 .838 .773 .414 .205 .623 .415 .437 .649 Sv-AA ppm > 0.080 .121 .091 .120 .121 .091 .110 .138 .105 .105 .105 .102 .104 .100 .128 .115 .101 .156 .174 .091 .134 .135 .151 .121 .135 Ftps scruni odu 0.0-0.3 Fecal Exam - i;s(f) _ .276 - - neg. -- - c(r); 1 -- --- -- - j1- -- .304 s(f) -- .302 s(0 ---- 1s(f) .334 s(D .390 - -- c(r);s(f) -- -- - c(r);s(m) -- -tCo Data Tables EID151729 a . c/1 to 53 I QW o o o to w w ti Tennant Farm Herd Health Investigation Cattle Team Report Individual Animal Data: Cattle from Tennant Farm (Washington, West Virginia) Table 4: Hematology (leukonj, Spet tai Chemistry, unti Fecal Exam (continued) Animal Age Preg. Number Status years | W ill' xloVul Neut Lymph Mono % % :;'o Eos % Huso % Prolactin plilMILI ng/ml Cu vertuti ppm Se-AA blmul ppm lVps scrum odu Fecal exam Reference 4-12 15-45 45-75 2-7 2-20 0-2 171.6 0.6- > 0.080 0.0-0.3 Range: 25 4 5 m 8.98 54.8 6.61 7.76 30.8 .026 (105.8) 47.76 1.20 .643 .161 -- 26 6 m 6.09 64.6 17.4 6.87 10.9 .168 27 >9 8 m 5.69 56.4 16.2 8.79 18.4 .160 68.95 .778 .142 .184 s (m) 122.27 .443 .116 - 28 7 8 m 5.92 69.9 19.1 .525 10.5 0.00 86.90 .396 .114 -- 29 >9 term 5.49 53.5 13.7 7.72 25.1 0.00 106.37 <0.20 .146 .224 neg. 30 >9 31 8 8m 6m 3.58 ... -6-4.-8- - 14.5 5.79 5.41 ~...i8.2.... ....3.61 15.0 __j 0.00 9.0 1 0.00 44.53 .289 .114 - 157.00 .683 .139 .098 - 32 >9 6m 6.86 59.0 19.8 3.36 17.8 0.00 361.97 .488 .132 - neg. 33 >9 8 m 6.76 70.4 4.69 6.72 18.2 .026 64.19 .533 .105 -- 34 >9 8 m 5.19 57.6 19.5 9.81 13.0 .056 109.60 .434 .161 .324 - 35 >9 36 >9 5m 8m 5.40 38.0 24.0 . 12.8 . --)3y - i 25.1 j .055 : 4.39 " '57~i... 17.5 |T . 9 ' 1 0.00 i 47.45 306.10 .679 .596 .140 .101 :- s (0 37 7 calf 5.89 54.0 14.4 10.2 21.3 .090 43.89 .732 .142 -- 00 >9 7 m 6.49 53.8 10.7 9.62 25.9 .022 143.13 .681 .124 -- 39 >9 calf 4.72 35.5 35.5 7.47 21.5 0 .0 0 48.70 .762 .109 qns - 40 4 6 m 9.05 26.7 29.7 12.6 30.9 .055 207.43 .607 .146 - s(f) 41 4 steer 9.57 52.3.... 31.3 2.73 13.7 0 .0 0 80.12 .582 .143 .304 Blood collected byjugularvenipunctureduring examination while cattle restrained in a head-hold device. Blood refrigerated for approximately 36 hours before hematology was run on an Abbott Cell-Dyne 3500. WBC (WIC values) considered valid; WBC differential (neutrophils, lymphocytes, monocytes, eosinophils, basophils) considered invalid due to 36-hour storage of blood. Hematologyreference values are for cattle (New Bolton Center), but not fromthe Cell-Dyne 3500. Seleniumvalues of 0.050ppmare considered marginal; below 0.050 ppmare seleniumdeficient. Prolactin (heparin tubesXDr. N. Schrick, Univ. ofTenn., Dept. An. Sei.). Ref. means: April non-fescue (171.6) and ergotamine tartrate (105.8 ng/ml). Cu =copper; Se-AA- selenium. (New Bolton Center) Peps =serumpepsinogen; odu - optical density units (Texas Veterinary Diagnostic Laboratory, College Station, TX) Fecal exam: c =coccidia oocysts; s = strongyle eggs; t - Monezia tapewormeggs; (r) = rare; (f) = few; (m) = moderate. (NewBolton Center),_____ Data Tables Tennant Farm Herd Health Investigation Cattle Team Report I oooo C/l Co 0CO o o o m D to W -uI $ Individual Animal Data: Cattle from Tennant Farm (Washington, West Virginia) Table 5: Clinical Chemistry A n im a l A ge P re g . IMJN C r t N a+ K+ C l- Ca Mg Phos Tot A lb (ilo b N um ber S ta tu s P ro t years ma/dl mg/dl mmol/1 inmo1/1 mmol/ mg/dl mg/dl nig/dl g/dl R/dl R/dl Reference 6 - 2 2 0.5 - 134 - 4 .0 - 9 6 - 8 .2 - 2 .0 - 4.7 - 5 .8 - 2 .4 - 3.0- Range: 1.1 144 5.7 104 10 2.8 9 .0 7.5 3.5 3.5 14 4 m 14.7 1.68 145 5.14 :0 2 9 .8 2.3 5.73 7.2 2.79 4.41 23 open 10.5 1.23 143 4.95 106 8.75 1.9 7.02 7.6 2.70 4 .90 3 >9 open 11.5 2.00 144 6.0 0 105 9.42 2.2 5.8 0 7.8 3.20 4 .60 4 >9 7 m 10.8 1.59 144 4.7 8 106 9.21 2.1 7.24 7.3 2.92 4.38 5 >9 6 m 7.9 1.82 146 5.06 107 7.71 2.0 9 31 7.8 2.96 4.84 6 >9 3 m 8.1 1.73 145 4.55 107 9.87 2.2 5.09 7.6 3.17 4.43 7 >9 6m 13.5 2.30 143 6.27 102 9.00 1.7 6.6 6 7.6 3.15 4.45 8 >9 open 10.0 1.47 145 6.13 106 9.58 1.9 5.3 0 8.0 3.03 4.97 9 >9 open 8.5 1.49 145 5.6 0 104 9.3 9 2.1 5.35 7.6 2.85 4.75 10 >9 term 17.0 2.15 150 4.91 109 9.93 2.2 6.11 7 .0 3.04 3.96 11 >9 8.5 m 11.6 1.86 149 4.81 111 10.24 ^ 2.3 5.02 ; 7.1 2,86 4.24 12 >9 term 21.0 1.62 ""7 4 6 '"'4 .6 4 109 " 9 .5 6 ' 1.7 "7 7 6 ! 7.6 2.86 4.74 13 >9 open 15.9 1.06 145 5.52 105 9 .1 0 1.8 7 20 ! 7.8 2.74 5.06 14 >9 calf 19.6 1.72 146 5.55 103 9.83 1.8 7.35 7.9 3.30 4 .60 15 >9 16 6 17 >9 Prcgn. open 4m 21.2 17.8 12.0 1.83 1.78 " 1.87 144 140 ....151 5.02 " 5.44 6.46 104 101 " 169" 8.42 2.0 6.37 9.44 2.1 . 5-~-49 7 0 .2 9 "__2 A__ 7.0 6.8 8.3 2.46 4.54 2.82 3.98 3.35 4.95 18 >9 4 m 8.2 1.73 144 4.85 102 9.52 2.0 6.76 7.9 3.42 4.48 19 >9 calf 14.3 1.69 147 4.72 105 9.34 2.0 5.07 7.1 3.01 4.09 20 >9 6m 11.3 2.09 146 5.36 107 10.32 2.1 5.95 8.1 3.07 5.03 21 >9 5 m 17.4 1.47 ' 146 " 4.77 " " IO S " 10.25 2.3 6.07 7.9 3.06__ 4.84 22 >9 5m 15.6 __2.09... .... 145 5.42 " "08.... 7 7 4 0 .. 2.2 " 7 9 3 .... 7.8 ' 3.25 4.55 23 - M l 8.5 2.55 ' 143 ~ 4.55 102 ' 9 . 4 2 " 2.2 j 5.55 7.6 3.08 4.52 24 >9 6 m 9 .6 1.98 148 4.67 110 8.19 I ! i!...7.03 " 7.5 3.03 4.47 AST IJ/L 58100 126 98 138 84 98 126 109 141 111 99 101 97 141 127 106 96 109 141 126 113 103 101 109 109 ( K c;C T U/L U/l. 22 64 89 __ 4 0 _ _ 77 3 6 "" 110 32 57 29 71 45 181 37 155 30 187 49 83 34 49 35 105 27 76 37" 97 42 86 35 52 34 85 30 87 31 298 39 79 28 92 35 96 30 9 0 34*" 97 33 116 37 Data Tables Tennant Farm Herd Health Investigation Cattle Team Report Individual Animal Data: CaLie from Tennant Farm (Washington, West Virginia) Table 5: Clinical Chemistry {continued) Animal Age Prog. HUN fre u t Na+ K+ Cl- Cu Mr Pitos Tot Alb (lini) AST CK CGT Number Status Pro! years I mg/dl mg/dl mmol/1 mmol/1 mmol/1 mg/dl mg/dl 3 1 g/dl g/dl g/dl U/L U/L U/L Reference 6 -2 2 0.5 - 134- 4.0- 96- 8.2- 2.0- 4.7- 5.8- 2.4- 3.0- 58- 22- Range: 1.1 144 5.7 104 10 2.8 9.0 7.5 3.5 3.5 100 64 25 4 5 m 10.0 1.99 149 4.93 108 9.90 2.4 7.12 7.7 3.34 4.36 122 138 29 26 6 M ! 7.6 2.08 147 4.69 107 9.72 2 2 5.31 7.7 3.13 4.57 139 150 54 27 >9 8 m 14.3 1.72 149 4.97 113 10.18 1.8 3.67 7.4 2.94 4.46 138 133 39~ 28 7 8 m 14.7 2.07 147 5.44 110 9.83 2.4 5.04 7.6 3.20 4.40 102 123 33 29 >9 term 12.9 2.16 146 5.26 109 10.19 2.2 5,35 7.3 2.83 4,47 98 50 37 30 >9 3l 8 8 m 14.1 164 ' 43 5.42 103 8.80 2.3 _6;77_ 7.5 2.99 4.51 132 90 rn6m II.3 L66 ... 48~... 4.55 109 i'33 2 ~ 2 ~ `~5.24'"' 7.3 3.25 ! 4.05 99 35 29 32 >9 6 m 18.8 1.72 143 4.36 106 9.80 2.2 4.72 7.3 2.98 4.32 89 126 43 33 >9 8 m 13.0 1.95 152 4.50 112 9.61 1.4 7.14 7.9 3.14 4.76 150 137 38 34 >9 8m 19.6 2.56 149 6.27 n o 10.79 2.5 6.75 7.9 3.52 4.38 139 99 35 35 >9 5 m 7.9 J . 8 I 148_ 5.77 - 107 9.82 j 2.3 5.30 8.0 3.27 4.73 120 106 30 36 >9 8 m 15.1 iTo9 ....147 "" 5 56 "" n o ... " 9.50 i 543 7.7 3.05 4.65 112 92 29 37 7 calf 9.8 1.92 146 5.55 106 9.43 2.0 4.12 8.2 3.43 4.77 119 94 36 38 >9 7 m 13.0 1.97 ..149..... 4.35 7 !2 9.65 2.1 5.34 8.2 3.47 4.73 206 173 39 O 39 >9 calf 6.4 2.03 145 4.90 102 9.44 1.9 5.31 8.1 3.01 5.09 135 79 33 o 40 4 6 m 15.0 1.94 147 5.05 108 10.20 2.2 6.28 7.6 3.12 4.48 147 79 37 o 4l 4 steer 8.3 1.72 143 6.37 106 ~9.95" 2.3 6.99 7.4 3.10 4.30 n o 192 26 o Blood collected by jugular venipuncture during examination while cattle restrained in a head-hold device. Blood refrigerated for approximately 36 hours C before serum chemistiy was run on a Kodak Ektachem. f r Cattle were corailed for several hours on a warm day, without water, before blood samples were collected. Reference values are for cattle (New Bolton Center, Kodak Ektachem), except for globulin (Duncan and Prasse). EID151732 W0 * Data Tables Tennant Farm Herd Health Investigation Cattle Team Report Individual Animal Data: Cattle from Tennant Farm (Washington, West Virginia) o oo C/l QCO o o o m o two -a o\ tCoO T able 6: S ero lo g y Animal Age Prig. ltl.V Johne's Brucella UVD BVD Blue l.i'plo Number Slatus Tongue tvrsl scrum lest: elisa elisa sn sn microplale AGID sn serum Reference samples Range: 14 4m neg neg neg neg neg neg 2 3 open neg neg neg neg neg neg neg 3 >9 open neg neg neg neg ... ."eg neg neg 4 >9 7m neg neg neg ___ neg neg neg neg 5 >9 6m neg neg neg neg neg neg neg 6 >9 3m neg neg neg neg neg neg neg 7 >9 6m neg neg neg neg neg neg neg 8 >9 9 >9 open open neg neg neg neg neg___ . nejL neg ...... neg__ _ neg neg neg neg neg neg 10 >9 term neg neg neg_ neg neg neg neg 11 >9 8.5 m neg - nc . ncS___ neg .... neg neg neg 12 >9 term neg neg neg neg neg neg neg 13 >9 open neg neg neg neg neg neg neg 14 >9 15 >9 calf pregn. neg __ _neg___ neg neg neg neg neg .... neg........ neg neg neg neg neg 16 6 open neg .... nee neg neg neg neg neg 17 >9 4m neg neg neg neg neg neg neg 18 >9 4 m neg ___ neg_._ n eg __ ; neg nejj neg neg 19 >9 20 >9 calf __ jneg___ _.... neg...... ___neg__ : neg 6 m neg _......neg___ neg __ .neg___ ___ neg___ neg neg neg 1:101) 2! >9 5m neg neg neg neg neg neg neg 22 >9 5m neg neg neg "eg neg neg neg 23 - bull neg ___neg__ neg ___ ?.?.&.___ ___ neg..... .....neS....J ___neg____ L _ J L _ >9 _ __6 m ___neg.... .....n e g ...... neg neg ___neg....... .... ...neg...... ' ___ neg___ FUI) (type 21 AGID neg neg - - - - - neg - >1 St) - neg neg - - - - ' v! Data Tables O C/T 53 Q CO OOO m 0 (0 01 0v)j -*4 * Tennant Farm Herd Healtli Investigation Cattle Team Report Individual Animal Data: Cattle from Tennant Farm (Washington, West Virginia) Table 6: Serology (continued) Animal Age Prig. BLV Johne's Hnii'i'lla nvn nvn Blue Lepto Mill) Number Stani'. urs) Tongue (type 2) f serum test: elisa elisa sn sn microplate AGID sn AGID serum samples Reference Range: i1 25 4 5m neg (neg)1 neg neg neg neg neg - 26 6 bull neg neg neg neg__ "g___ neg neg neg 27 >9 8m POS neg neg 1:256 neg neg neg - 28 7 8m neg neg neg neg neg neg neg - 29 >9 term neg ___neg.... . .....neg____ ......neg___ neg neg 1-40(1 >1 SO 30 >9 8m neg neg neg neg neg neg neg - 31 8 6m neg neg neg neg neg neg neg neg 32 >9 6m neg neg neg neg neg neg neg - 33 >9 8m neg neg neg neg neg neg 1:100 - 34 >9 8 m neg neg neg ___ neg___ neg neg neg neg 35 >9 36 >9 5m 8m neg neg neg. neg neg neg ... neg neg neg___ __ neg___ neg neg neg neg - 37 7 calf neg neg neg neg neg neg neg - 38 >9 39 >9 40 4 7m calf 6m neg neg neg neg neg neg neg ___ QS___ ___ qs___ ; - neg __ neg____ neg ____eg neg ___ neg neg POS neg neg neg neg neg - 41 4 steer neg BLV = bovine leukemia virus antibody neg neg . neg . neg neg neg Johne's = Mycobacterium paratuberculosis antibody (technique has a 1% false positive error rate) Brucella = Brucella abortus antibody Lepto = antibody to Leptospira spp. (Lpomona, L icterohemorrhagiae, L hardjo, L grippotyphosa, L canicola). BLV, Johne's, Brucella, and Lepto assays conducted by the Animal Diagnostic Laboratory, Penn. State Univ., University Park, PA. EHD = 2 of 12 tested cattle were ``positive" for EHD-type 2 (Alberta strain); (Texas Veterinary Diagnostic Laboratory, College Station, TX). Data Tables Tennant Farm Herd Health Investigation Cattle Team Report Tennant Farm (Washington, West Virginia): Grain and Hay Analysis Tabla 7: Nutritional Analysis Hay Grain Analyte units as sampled dry matter as sampled dry matter . Dry Matter basis basis % 86.34 basis 89.66 basis --I Crude Protein % 6.77 ........ 7JB4...... 10.21 ..... 11.39 .... Unavailable Protein % 1.90 2.20 - Adj. Crude Protein % 5.55 6.42 - - Acid Detergent Fiber TDN NE (Lactation) Calcium Phosphorus % % Mcal./lb ppm ppm __ 40.44 46.84 47.67 55.21 0.33 ...... ....... 0.38 ~ i i 0.40 0.46 0.10 0.12 !""1 7.59 69.35 0.72 0.95 0.30 8.46 77.35 0.81 1.07 0.33 Sodium Magnesium Sulfur Potassium % 0.02 0.02 %. 0.13 0.15 % 0.18 0.21 % 0.87 1.01 0.37 0.48 0.30 0.34 0.13 0.14 0.60 0.67 Copper Iron ppm 5 6 ppm 206 239 67 305 340 Manganese Zinc Nitrate Ion Selenium PP1 PPm % ppm _ 331 43 0.00 384 50 0.00 0.12 22 24 39 43 r- 0.19 Hay and grain feed analysis performed by NC Dept. Agric., Food and Drug Protection Division, Forage Testing Laboratory, 4000 Reedy Creek Rd, Raleigh, NC 27607. Hay and grain samples collected by cattle team during April 1999 site visit to Tennant Farm. Hay sample consisted of 3 _ core samples each from 3 large round bales of 1998 cut hay. Data Tables Tennant Farm Herd Health Investigation Cattle Team Report 12.0 APPENDICES Appendix A: Abbreviated Curriculum Vitae of Cattle Team Members Appendix B: Diagnostic Pathology Reports Ohio Department of Agriculture: #4977-97 Michigan Animal Health Diagnostic Laboratory: #1792571 University of Pennsylvania Laboratory of Large Animal Pathology and Toxicology: #UP9902702, #9901437 Appendix C: Dry Run Safety Plan Appendix D: Figures 1-42 : Photographs of the 42 adult cattle; Figures 43-66 : Miscellaneous photographs of the Tennant Farm and cattle Appendix E: Herd Health History (April 8, 1999 interview) Appendix F: Diet Analysis: Computer Software Modelling 000058 sr EID151736 R G S000939 Tennant Farm Herd Health Investigation Cattle Team Report Appendix A: Abbreviated C u r ric u lu m V itae of the Cattle Team Members Name: Degrees, Certifications: Employment: Education; Dr. Perry L. Habecker VMD. Diplomate ACVP Chief. Large Animal Pathology. Laboratory of Pathology and Toxicology. Univ. of Penn. School of Veterinary Medicine, New Bolton Center. Kennett Square, PA. Juniata College (BS. I972-I976) University of Pennsylvania. School of Veterinary Medicine (VMD 1969-1973) University of Pennsylvania. School of Veterinary Medicine (resident, 1989-1992) Affiliations: American College of Veterinary Pathologists Pennsylvania Veterinary Medical Association American Association of Veterinary Laboratory Diagnosticians Name: Degrees. Certifications: Employment: Education: Affiliations: Dr. Lisa Davis-Heller DVM Private Practitioner. St. Mary's Veterinary Clinic. St. Mary's. WV. Ohio Slate University Ohio State University, College of Veterinary Medicine (DVM. 1979-1983) American Veterinary Medical Association Ohio Veterinary Medical Association West Virginia Veterinary Medical Association International Veterinary Acupuncture Society Holistic Veterinary Society Name: Degrees. Certifications: Employment: Education: Affiliations: Dr. Peter G. Moisan ** DVM. Diplomate ACVP.. Diplomate ABVP (food animal specialist: beef cattle specialist) Veterinary Pathologist and Field Investigator. Rollins Animal Disease DiagnosticLaboratory. North Carolina Department of Agriculture. Raleigh. NC. Cleteland State Community College (AS. 1974) East Tennessee State University (BS, Microbiology, 1975) East Tennessee State University (MS. Microbiology, 1978) University of Tennessee, College of Veterinary Medicine (DVM, 1981) Michigan State University (Resident, Veterinary Pathology, 1995) Kansas State University (Research. Veterinary Pathology, 1995-1998 American Board of Veterinary Practitioners American College of Veterinary Pathologists American Association of Bovine Practitioners American Association of Swine Practitioners Academy of Veterinary Consultants American Veterinary Medical Association American Association of Veterinary Laboratory Diagnosticians_______________ Appendix A: Abbreviated C urriculum Vitae of Cattle Team Members 000059 S<o EID151737 R G S000940 Tennant Farm Herd Health Investigation Cattle Team Report Name: Degrees, Certifications: Employment: Education: Affiliations: Dr. Robert J. Munson VMD Field Investigator, Center for Animal Health and Productivity, Univ. of Penn. School of Veterinary Medicine, Kennett Square, PA. Dickinson College (BS, 1963-1967) University of Pennsylvania School of Veterinary Medicine (VMD 1969-1973) American Veterinary Medical Association American Association of Bovine Practitioners American Dairy Science Association Name: Degrees, Certifications: Employment: Education: Affiliations: Dr. Robert H. Poppenga DVM, PhD, Diplomate, ABVT Chief, Toxicology, Laboratory of Pathology and Toxicology, Univ. of Penn. School of Veterinary Medicine, New Bolton Center, Kennett Square, PA. Western Illinois University (biological sciences, pre-vet medicine, 1971-1974) University of Illinois. College of Veterinary Medicine (DVM, 1974-1978) University of Illinois. College of Veterinary Medicine (PhD, 1983-1987) American Veterinary Medical Association Pennsylvania Veterinary Medical Association American Academy of Veterinary and Comparative Toxicology American Academy of Clinical Toxicology Society of Environmental Toxicology and Chemistry Society of Toxicologic Pathologists Society of Toxicology American Association of Veterinary Laboratory Diagnosticians Name: Degrees. Certifications: Employment: Education: Affiliations: Dr. Greg P. Sykes VMD, Diplomate ACVP, Diplomate ABT Pathologist. Safety Assessment, DuPont Pharmaceuticals Company (wholly- owned subsidiary of the DuPont Co.), Stine Research Center, Newark. DE. Cornell University: (biological sciences; 1968-1971); University of Pennsylvania School of Veterinary Medicine (VMD 1971-1975) Cornell University. NYS College of Vet Medicine (resident/fellow, 1979-1983} American Veterinary Medical Association American College of Veterinary Pathologists International Academy of Pathology C.L.Davis Foundation for the Advancement of Veterinary Pathology Royal Microscopical Society American Board of Toxicology American Association of Laboratory Animal Science Association of Wildlife Veterinarians Appendix A: Abbreviated C urriculum Vitae of Cattle Team Members 000060 S '7 EID151738 R G S000941 REPORT OF LABORATORY EXAMINATION ANIMAL HEALTH DIAGNOSTIC LABORATORY P.O. Box 30076 Lansing. Ml, 48909 Phono (517) 353-5275 PRIVILEGED INFORMATION NOT FOR PUBLICA TION ^ nm ^ F IN A L FACH 1 OP S GROSS NECROPSY C asa N ustber: R ep o rted R eceiv ed P a th o lo g ist : C ass O rig in : 1792571 0 3 /1 2 /9 7 0 3 /0 4 /9 7 JS P NECROPSY C lie n t A cco u n t: 258989 C lin ic : Jx) CA SPA R, SARAH U .S . BPA REG. 3 8 41 CHESTNUT BUILDING PHILADELPHIA PA 1 9 1 0 7 HISTORY : M any c a t t l e o n o n e f a r m ( o v e r ISO o u t o f 3 0 0 ) h a v e d i e d o v e r t h e p a e t y e a r. C lin ic a l s ig n s in c lu d e b lack en ed te e th , p a tc h y h a ir lo s s an d g ra y in g o f th e h a ir c o a t, lo s s o f h a ir in th e t a i l , o v erg ro w th o f th e ho o v es, and w eig h t lo s s . N ecropsy fin d in g s in c lu d e d p a tc h y , b la c k d is c o lo ra tio n o f th e ru m in a l m ucosa and p o s s ib le a b n o rm a litie s in th e k id n e y s. C lin ic a l s ig n s and n e cro p sy fin d in g s w ere d e s c rib e d in a v id e o ta p e w hich w as s u b m itte d w ith th e tis s u e s . T is s u e s from a 4 y e a r - o l d c o w (AKDL 1 7 9 2 5 7 1 - 1 ) w h i c h d i e d o n 2 / 1 8 / 9 7 a n d t i s s u e s f r o m a 9 - y e a r - o l d c o w (AHDL 1 7 9 2 5 7 1 - 2 ) w h i c h d i e d o n 3 / 2 / 9 7 w e r e su b m itte d . E xposure to an e n v iro n m en ta l c o n tam in an t i s s u s p e c te d . A nim . # : 1 Name: 2 /1 8 B r e e d : HOLSTEIN Age: 4y S e x : FEMALE LABORATORY FIN D IN G S TOXICOLOGY RESULTS AND COMMENTS SPECIM EN: LIVER T e s t : T IS S U E MXNBRAL AN ALYSIS (v a lu e s i n ppm ) B ( < 1 .0 0 Co ( < 0 .1 0 0 Mo ( 0 .8 8 6 AS ( < 0 .5 0 0 Pb ( < 0 .5 0 0 K < 2070 ) Ba ( < 0 .1 0 0 ) Fe { 170 ) P ( 2850 ) C r ( < 0 .2 0 0 ) Se ( < 2 .0 0 ) ) Ca ( 5 1 .7 ) Mg ( 1 56 ) Zn ( 4 3 .2 > Cd 1 0 .1 3 0 ) T1 ( < 2 .5 0 ) Cu ( 1 .9 0 ) Ma ( 1 .5 3 ) Sb < < 1 .0 0 ) Hg ( < 2 .0 0 ) Na ( 1360 ) ) ) ) ) T est C om m ent: C opper i s w ith in d e fic ie n t ra n g e . C opper d e fic ie n c y cau ses a c u te d e a th and d is c o lo r a tio n o f h a ir . M anganese i s m a rg in a lly lo w . O th er tis s u e e le m en t c o n c e n tra tio n s a re w ith in e x p e c te d ra n g e s. WXR 0 3 / 0 7 / 9 7 *DENOTES ADDm ONA l TEST RESULTS ____ MSU IS AN AFFIRMATIVE ACTION/EQUAL OPPORTUNITY INSTITUTION 000061 53 EID151739 RGS 000942 F IN A L PAGE 2 OF C ase N um ber: 1792571 5 (1) SPECIM EN : KIDNEY T e a t : T IS S U E MINERAL ANALYSIS (v a lu e s in ppm ) B ( < 1 .0 0 Co ( < 0 .1 0 0 Mo ( 0 .3 3 4 A s ( < 0 .5 0 0 Pb ( < 0 .5 0 0 K ( 2040 ) Ba ( 0 .2 0 1 ) Fe ( n a ) F ( 2230 ) C r ( < 0 .2 0 0 ) Se ( < 2 .0 0 ) ) Ca ( 9 3 .5 ) Mg ( 169 ) Zn ( 1 9 .8 ) Cd ( 0 .8 3 8 ) T I < < 2 .5 0 ) Cu ( 2 .3 7 ) Mn ( 0 . 6 0 5 ) Sb ( < 1 .0 0 ) Hg C < 2 .0 0 ) Na ( ISSO ) ) ) ) ) T eat C om m ent: C o p p er i# below n o rm al ra n g e (< 0 .5 ppm). WKR 0 3 / 0 7 / 9 7 ra n g e (4 -6 ppm) a n d cadm ium i a a b o v e n o rm a l SPEC IM EN : URINE T e s t : T IS S U E MINERAL ANALYSIS (v a lu e s in ppm ) B ( 2 .9 7 ) Ba ( 0 .9 5 6 Co ( < 0 .0 5 0 0 ) Fe ( 0 .5 2 3 Mo ( < 0 .1 0 0 ) P ( 7 6 .2 As ( < 0 .2 5 0 ) C r ( < 0 .1 0 0 Pb ( < 0 .2 5 0 ) Se ( < 1 .0 0 K < 6430 ) ) Ca { 409 ) Mg ( 4 2 8 ) Zn ( 0 .6 3 6 ) Cd ( < 0 .0 5 0 0 ) T I ( < 1 .2 5 ) Cu ( 0 .0 6 5 0 > Mn ( < 0 . 0 2 5 0 ) Sb ( < 0 .5 0 0 ) Hg ( < 1 .0 0 ) Na ( 240 ) ) ) ) ) T e s t C om m ent: No heavy m etal fo u n d in th e u rin e a t th e re p o rte d d e te c tio n lim i ts . WKR 0 3 / 0 7 / 9 7 B o th l i v e r s an d k id n e y s had low c o p p e r. T h is i s th e m a jo r f in d in g so f a r . R e p o rte d a d eq u a te l iv e r m in e ra l ra n g e s f o r c a t t le a re : c a lc iu m 30 t o 200 ppm; c o p p e r, 25 to 150 ppm ; i r o n , 45 to 300 ppm ; m a g n e s iu m , 1 0 0 t o 2 0 0 p p m ; m a n g a n e s e , 2 . 5 t o 4 . 0 ppm,- m o ly b d e n u m , 0 . 1 4 t o 1 . 4 0 ppm ; p h o s p h o r u s , 1 5 0 0 t o 4 1 0 0 ppm,- z i n c , 2 5 t o 2 0 0 ppm ; so d iu m , 600 to 1900 ppm ; and p o ta e siu m , 1200 to 3300 ppm. T o x i c a n t s c r e e n b y GC/MS o n t h e l i v e r i s i n p r o g r e s s a n d w i l l b e re p o rte d o u t in a su p p le m e n tal r e p o rt when a v a ila b le . N u tr itio n a l e x a m in a tio n : U rin e f lu o r id e c o n c e n tra tio n waa 6 .6 u g /m l. A nim . 2 Name: 3 /2 B r e e d : HOLSTEIN A ge: 9y S e x : FEMALE ` DENOTES ADOm ONAi TEST RESULTS 000062 SV EID151740 R G S000943 P IN A L PAGE 3 OF C ase N um ber: 1792571 5 {1, SPSC IH SN : FIXED TISSU ES HISTOPATHOLOGIC EXAMINATION S e c tio n s o f h e a r t, l i v e r , an d k id n e y w ere ex am in ed . In s e c tio n s o f l i v e r , th e r e waa ad v an ced p o stm o rte m a u to ly s ia , w ith fre e z e /th a w a r t i f a c t s in one s e c tio n . M oat h e p a to c y te a c o n ta in e d s in g le , sm a ll, ro u n d , c le a r, in tr c y to p la sm ic v a c u o le s , in d ic a tin g m ild , d iffu s e h e p a tic lip id o s is . In s e c tio n s o f k id n ey , sw at g lo m e ru li c o n ta in e d s l i g h tl y in c re a s e d q u a n titie s o f e o s in o p h ilic m esan g ial m a trix . The s i g n i f ic a n c e o f t h i s f in d in g i s unknow n, a n d i t may b e a p o stm o rte m a r t i f a c t . In s e c tio n s o f h e a r t, sm a ll num bers o f m y o fib ere c o n ta in e d S a rc o c y stis c y sts. LABORATORY FINDINGS TOXICOLOGY RESULTS AND COMMENTS SPECIM EN: LIVER T a s t T IS S U E MINERAL ANALYSIS ( v a lu e s in ppm ) B ( < 1 .0 0 Co ( 0 .1 4 1 Mo ( 0 . 7 5 8 Aa ( < 0 .5 0 0 Pb ( < 0 .5 0 0 K < 2670 ) Ba ( < 0 .1 0 0 ) Fe ( 356 ) P ( 2810 ) C r ( < 0 .2 0 0 ) Se ( < 2 .0 0 ) ) Ca ( 3 2 .7 ) Mg- < 149 ) Zn ( 8 8 .6 ) Cd ( 0 .2 1 9 ) T l ( < 2 .5 0 ) Cu 2 .0 3 ) Mn 1 .7 6 ) Sb { < 1 .0 0 ) Hg < 2 .0 0 ) Na 1040 T e a t C oirm ene : C opper ia w ith in d e f ic ie n t ra n g e . The fo llo w in g e le m en t(a) b elo w n orm al ra n g e (a ) : m an g an ese. The fo llo w in g e le m e n t(s) a b o v e n o rm al ra n g e (a) : I r o n WKR 0 3 / 0 7 / 9 7 ia /a re is/a re SPEC IM EN : KIDNEY T e a t T IS S U E MINBRAL ANALYSIS (v a lu e s in ppm ) B ( < 1 .0 0 Co ( < 0 .1 0 0 MO ( 0 . 3 8 8 AS ( < 0 .5 0 0 Pb ( < 0 .5 0 0 K ( 2100 ) Ba { 0 .3 2 7 ) Fe ( 7 1 .5 ) P ( 1880 ) C r ( < 0 .2 0 0 ) Se ( < 2 .0 0 ) ) Ca ( 6 3 .7 ) Kg ( 126 ) Zn ( 2 6 .5 ) Cd ( 1 .7 6 ) T l ( < 2 .5 0 ) Cu ( 2 .3 3 ) Mn ( 0 .5 9 5 > Sb ( < 1 .0 0 Kg < 2 .0 0 ) Na ( 2020 DENOTES AOOmONAL TEST RESULTS MSU IS AN AFFIRMATIVE ACTION/EQUAL OPPORTUNITY WSTITUTION 000063 60 __ EID151741 R G S000944 F IN A L PAGB 4 OF C ase R um bar: 1792571 5 (1) SPECIMEN : HAIR T e s t : GENERAL MINERAL ANALYSIS (v a lu e s in ppm ) B ( < 4 .0 0 Co ( 0 .5 1 7 Mo ( < 0 .8 0 0 A s ( < 2 .0 0 Pb ( < 2 .0 0 K ( 973 Ba ( 9 .3 7 Fe ( 263 P ( 288 C r < < 0 .8 0 0 Se ( < 8 .0 0 ) ) Ca ( 1740 ) Mg ( 1 0 9 0 ) Zn ( 7 1 .8 ) Cd ( < 0 .4 0 0 ) T1 < < 1 0 .0 ) Cu < 7 .2 3 ) Mn ( 2 9 . 9 ) Sb ( < 4 .0 0 ) Hg ( < 8 .0 0 ) Na ( 6 8 .4 ) ) ) ) ) SPECIM EN: URINE T e s t : FLUID MINERAL ANALYSIS (v a lu e s in ppm ) B 2 .7 0 CO < 0 . 0 5 0 0 Mo < 0 . 1 0 0 As < 0 .2 5 0 Pb < 0 .2 5 0 K 8400 Ba ( 0 .3 2 8 Fe ( < 0 .2 5 0 P ( 131 C r ( < 0 .1 0 0 Se < < 1 .0 0 ) Ca ( 1 8 .2 ) Cu ( < 0 .0 2 5 0 ) ) Mg ( IBB ) Mn ( < 0 .0 2 5 0 ) ) Zn ( 0 .1 0 1 ) Sb ( < 0 .5 0 0 ) ) Cd ( < 0 .0 5 0 0 ) Hg ( < 1 .0 0 ) ) T1 ( < 1 .2 5 ) Na ( 231 ) B oth liv e rB and k id n e y s h ad la v c o p p e r. T h is i s th e m a jo r f in d in g so f a r . R e p o rte d a d eq u a te liv e r m in e ra l ra n g e s f o r c a t t l e a r e : calciu m 30 t o 200 ppm ; c o p p e r, 25 to 150 ppm ; ir o n , 45 t o 300 ppm ; m ag n esiu m , 100 t o 200 ppm ; m a n g a n e se , 2 .5 t o 4 .0 ppm ; m olybdenum , 0 .1 4 to 1 .4 0 ppm ; p h o sp h o ru s, 1500 to 4100 ppm ; z in c , 25 to 200 ppm; sodium , 600 to 1900 ppm ; and p o ta s s iu m , 1200 to 3300 ppm . T o x i c a n t s c r e e n b y GC/MS o n t h e l i v e r i s i n p r o g r e s s a n d r e s u l t s w i l l be re p o rte d in a su p p le m e n tal r e p o rt when a v a ila b le . N u tr itio n a l e x a m in a tio n : U rin e flu o r id e c o n c e n tra tio n was 5 .5 5 u g /m l. SPEC IM EN : SERUM C L IN IC A L PATHOLOGY Serum c h e m is try p r o f i le w as ru n on a sam p le s u b m itte d from cow # 2 . A b n o rm a litie s in c lu d e d s l i g h t l y low sod iu m c o n c e n tr a tio n (1 3 7 .6 m m ol/L ; r e f e r e n c e ra n g e - 1 3 9 -1 4 5 ), s l i g h t l y low so d iu m t o p o ta s s iu m r a t i o (2 4 ; r e f e r e n c e ra n g e 2 5 -3 5 ), low seru m a lb u m in c o n c e n tr a tio n (2 .9 g / d l ; r e fe r e n c e ra n g e 3 .5 - 5 .0 ) , s l i g h t l y low alb u m in to g lo b u lin r a ti o (0 .8 1 ; re fe re n c e ra n g e 1 .0 1 -3 .3 9 ), s lig h tly e le v a te d t o t a l b i l i r u b i n < 0.4 m g /d l; r e f e r e n c e ra n g e < 0 .3 ) , s l i g h t l y e le v a te d c r e a ti n i n e (1 .7 m g /d l; r e f e r e n c e ra n g e - 0 .6 - 1 .3 ) , low c a lc iu m c o n c e n tra tio n (6 .5 m g /d l; r e fe re n c e ra n g e 7 .9 -1 0 .7 ), s l i g h t l y low s o r b i t o l d e h y d ro g e n a s e a c t i v i t y ( 8 .6 UL; r e f e r e n c e ra n g e 1 3 .4 - 7 0 .7 ) , s l i g h t l y e le v a t e d se ru m i r o n c o n c e n tr a t io n (138 u g / d l ; r e f e r e n c e ra n g e - 4 0 -1 3 6 ), a n d s l i g h t l y lo w o s m o l a l i ty (283 m O s/kg; re fe re n c e range 286-302). 6/ DENOTES ADDITIONAL TEST RESULTS MSU IS AN AFFIRMATIVE ACTION/EQUAL OPPORTUNITY INSTITUTION 000064 EID151742 R G S000945 FIN A L PACK 5 OP C aae N um ber: 1792571 S (I) COMMENTS: No a ig n if ic a n t p a th o lo g ic fin d in g . [T o x ic o lo g ic a n a ly a e a a re in p ro g re a a an d a a u p p le m en tal r e p o rt w ill # b e B en t w hen r e a u l ta a re a v a i l a b l e . R . P u la in 'M in e ra l L e v a la i n A nim al H e a lth * g iv e a th e n o rm a l le v e l o f f lu o r id e i n c a t t l e u r in e a a 0 .7 to 5 .0 ppm, th e h ig h le v e l a a 5 .0 to 1 5 .0 ppm, a n d th e to x ic l e v e l a a 1 4 .0 - 1 2 0 .0 p p m .] Jo n S. P a tte ra o n P a th o lo g ia t }ma (517) 353-5275 9 * * t * % t l * * * % % l DENOTES ADDITIONAL TEST RESULTS MSU IS AN AFFIRMATIVE ACHON/EQUAL OPPORTUNITY WSTTTUTION EID151743 000065 R G S000946 FRCM : BEURRE P H I*. CLINIC F K * e M3. : 614 423 1750 3A W A n im a l D ise u se Diogn* ' v Kr f 'n tto r y -- 2 -J L - c ~ % pe -X 2 S * % A tam lo n : Rpori data: O w n: Coordinator: 4 9 7 7 -8 7 J-10-97/SC Cuatar. l n 0 M 7-6135 Dr. S la b Grin .aa BELPRE. OH 48714 Ohio Caparimant ol Agrtovihura 8999 East Main St. Reynoldsburg. O H 1 3 0 6 8 6 1 4 -7 2 8 -6 2 2 0 FAX: 728-0310 SUBM tSSJON SUM M ARY Taken: not given Received: 2 26 87 Speolee Bovine Animala Tecta Completed 18 8 PATHOLOGY ADOCNDCD MEFOHT ANIM AL ID: 1: (not provided). Bovine. M ixed B reed B eef Sample: Doad Animal N E C R O PSY 4 H'STOPATHOLOCY 2 -2 7 -9 7 (1 1 :2 4 N ecropsy: H istory: Cp M O X d e a d . a iv -m o o th -olH h u t c all w a s s u b m i tt e d . T h e c a l f ta u n d e rw e ig h t a n d h a s h a d d ii. ih e a lo r o n m o n th . A ccording lo Ih h k to ry , tHe calf h a s n o t b e e n dow orm ed end hae n o t racu rad any m ed icatio n . The aye ol the cow w ere o b serv ed to h a v e oloudy earner*. Grose examination: T h e m ale oaH w ac presented dead. T he c a rc a s s w tlg h M 1SO U . The nutritional s ta ti v a s poor. M inim a) b o d y fat w a s p re s e n t o n t h e c a r c a s s a n d s e r o u c a tr o p h y e l fa t w e e p r e s e n t a/> u n d th e h e a r t a n d k id n ey s. T he s ta te of h y d ra tio n w a s fair. A ta w T n c ttu m w e re p ro ean t in th e e e c tu n . H ay. b u t no grain w as p re ca n t In th e fo resto m ac Ire. Form ed facet belie w e re p u m in t h e r e d u rn . F e o e s w e r e m a n e d e ro u tid t h e a n u s .. M i d f o a m w a s p r e s e n t in th e tra o h e .1 N o additional g ross lesions w ere observed. M orphologic diagnosis: E m aciation w ith ssrous atrophy of fat In te stin a l parasitism (trichuriasis) Comments: T h e c a l f w a s in an e x tre m e ly p o o r n u tritio n a l s t a l e . Outing in e te m e m w e a th e r e e n d ltlo r C e . . c o l d a s s o c i a t e d w ith w in te r) n u tritio n a l r e q u ir e m e n ts e f e n im e la in c re a s e a n d it is e x tr e r r >ly im p o r ta n t tn irw.rrunta th e feed in tak e e t t h e a n im a ls . H e y , b u t n o grain, w s s o b s e rv e d in th e f o r e s t o m a c h s . S u p p le m e n ta tio n e f t h e h a y w i t h a d d itio n a l fo o d c o u re o c m a y b e b e n e fie I. A d d itio n a lly , ssv ara l parasite ova w e re o b se rv e d in th e f e c e s an d w hipw orm * w ere o b e* v e d in th e c e c u m . A p a rasitic burden raduesa th e ttvo n u tritio n al in ta k e o f tn arw nal, com pound; th a 000066 0 EID151744 R G S000947 FROM : BELPRE PNIflFL CLINIC P W > M3. : 6 1 4 4 2 3 1750 POl A eoeealon: A S7747 t c e i l t l n u a d ) ------ V stadnariw . A Joway. Clyde H. Jr. O w m r C u tter. K avh 1 '"= = = 1 1 ....... ......... -- --. Page: 2 NECROPSY & HISTOPATHOLOQY 2-27-57/11:24c (Continued) pravioualy pear nutritional atata. Histopathologic. baeteriologic and virologlc aaaminationa ara _currently In pregreee. Laalana obaerved in the bulPa eyeamary have bean a reeult efthu eold, but a hktopathelogic examination af tha ayea la eurrantiy in progress. Sheila Grimes. OVM. PhO. Oiplomata. ACVP Veterinary Pathologist Pathology Section Hiatopathologic axaminatien: Sootlont of earbrum, cerebeSum. brslnstam. king. lymph node, k w , Udnay. adrenal, duedonum. jejunum. ileum. colon, heart and ebelatel m a d e were examined. Moderate a aevera paatmortam autolyala waa present In the aaettona af Intestine. Several ooonldbl eohlseitta. ranging In atea from 200 to 500 mloroiie In diameter, ware praaam In the emal bnextfcval mueeco. W linln tha aaction of colon, ariruat atagea n l cnrcidia. including caocidial aaaya. gamenta and gnmataa. * ware eraeant within orypt aphhalal oalW. Crypt abaaaaaaa ware praaam within aoma a- Ionia erypta. The orypu war* atari and dried by attenuated aplthadum. Tha eoetlen at ttvar had mild ta mederxia. diffuaa micieveaieular vacualatien of hepalacytav. Hepetocytee eantainsd m ultiple, poorly daBnaatad vaeuolec withIn thaIf ayteplaama. Ctiologic diagnoaia: Enteric oocoldleei sf ft Shaila C rim ea. OVM. PhO. Oiplomata. ACVP V eterin ary Patholofliat Pathology Saetlon Hiatopathologic examination: T wo aoctlona of eye ware examined. The aye had a chronlo karathia leornoal aaer) ahi racterlted by tha precence of a foealy extnaive area of flbropieela consisting af ralathraiy leaaali arranged fibrobleata artmiml with a few tymphocytaa and plaeme ceNe in tha eemae. 4Hd vascularization and edema was preaent in the araaa of flbroare. The cemeei ephhelVum overlying the area o l fibroek waa hyperplaatio and undergoing apidarmal metapUaia whh the formation of rate ridge. Oeacamot's membrane waa cut and retracted leaded). M orphologic diagnosis: M odorats. chronlo. focefly axtonahro fibreelng karatkic Sheila Grime. OVM. PhO. Oiplomata. ACVP Veterinary Pathologist Pathology Section 000067 & EID151745 R G S000948 FRCn: B8LPREANInPLCLINIC not to. :6144Z31750 A-- Jon: M 7 7 4 7 Voesrtoarian: A iow ey. Clyd H. Jr. (oOntJniMd) *-: i- I -- ........ -- a t u t u - j <mmr. C u n er. Kevin go: 0 P02 FECA L FLOTATION 3-37-C7/11:34o (Continued) FIC A L FLOTATION 2-37-7/11:24 R esulti: Numerous suongyle type egge. moderne numbers el oooeidUl ooc y t e and a few whipworm egge wot* sb n rrrd . ANIMAL IO: 1: (not provided), bovine. Mixed Creed | e l SALM ON Ft t A CUt TURF 3-3-87/1 1:24e Resulta: Negative ' D IA G N O ST IC VIRO LO G Y ANIMAL IO: 1: (not provided), levine. Mixed Creed Beef . FA -CO RO N A VIRUS 2-27-97/4;C1p Results: Negative ANIMAL IO: 1: (not provided). Bovine, Mixed Breed Boo/ FA-CORONA VIRUS 2-27-*7/4:9tp Results: Negativo FA-OIAROlA 2-27-87/4:81 p AeaulU: Negative FA -C R Y FT O S PORIDIA 2-27-7/4:91p Resulta: Negative F n p ls t Pooled Tissue Culture Compie: Intanine Compia: Ileum 000068 bS EID151746 R G S000949 . FSQM:BELPIPN1MPLCLINIC FKNO. :6144231750 A ccessio n : 4677-67 Veterinarian: A*oway. Clyde H. Jr. Ownan Cuatar. Kevin FINAL DIAGNOSIS 4 COMMENTS S-4-S7/5i4Cp (Continued) PATHOLOGY AN IM AL 10: 1: Inal provided). tavina. Mbcad Braad Baaf FIN A L D IAGN O SIS 4 COMMENTS S-a,S7/ti40p DIA G N O SIS em aciation Atrophy. Saroua Ceoeldloele, Enteric T ric h u ria s is Keratitis tempi: Daad Animal Comments: Please refer to iha previous commanta. The acoura obaarvad in tha animal waa probably a raault of enteric caccidioaia. Hielopsthelogic anamination of tha eya a currently in progress and results will be reported in an addendum. Shelia Grim#. DVM. PhO. Diplomats. ACVP Veterinary Pathologist Pathology Section Addendum: Plaeea refer to the histopathologic description of the aye. leeiene in tha eye were inda ctiao o f o prior oornoal Injury Cornoal Injury may result from physioal or ehemloal trauma, m icrobial agarrta. Inoraaaad ocular proaaura and taraly. Inborn errors of motaboliom. * ha exact cause of tha karetitia or cornoal wound in this case waa not determined. Lesions -n the eye ware euggeetiva of theca associated wkh trauma, but a Vaumatlo Injury in both ayes would be unuausl. Tha ulcerating and suppurative karatitla associated with AforuoOe waa net obaarvad In thin case. Howavar. fibrosis may occur with healing subsequent to tha inftetien. Sheila Crim ea. DVM. PhD. Diplomats. ACVP Veterinary Pathologist Pathology Section 000069 u EID151747 R G S000950 NEW BOL TON CENTER University of Pennsylvania, New Bolton Center Laboratory of Large Animal Pathology and Toxicology 382 W. Street Road Kennett Square, PA 19348 Phone: (610)444-5800 Fax: (610) 925-8110 FINAL REPORT Accession No.: UP 9902707 Submitter Dr. Perry Habecker Pg* Report Address: Dry Run Investigation Team Case Tracking ft: N o p. Case Coordinator Perry L. Habecker, VMD Date Submitted: 6/27/99 Report Date: 7/5/99 TA(D): 8 Species: Bovine Production type: Breed: Sex: F Age:__ l y Animal I D. Sample: .... 14..... Fixed Tissue Date obtained: Reference Lab: m w d Adult O Fetus O Juvenile O Unknown HISTORY SUMMARY: Killed and necropsied for Dr. Lisa Heller on June 10, 1999. Tissues also submitted to toxicology (ADDENDUM: tissues for toxicologic assay were received 7/28 and accessioned as 9903263) DIAGNOSIS: Enteric lesions of minima] significance. COMMENTS: The gastrointestinal eosinophilia is attributed to previous and current bouts of endoparasitism. Epithelial abscessation in the forestomachs is associated with acidosis due to excessive carbohydrate ingestion. LABORATORY FINDINGS: H1STOPATHOLOGY EXAMINATION 1) Heart, 2 pieces: Minimal, focal lymphocytic myocarditis', focal Sarcocyst spp.. 2) Lung, 2 pieces: Normal 3) Liver, 2 pieces Normal 4) Spleen: Normal 5) Kidney: Normal 6) Teat Normal 7) Adrenal, 2 pieces: Normal 8) Lymph node, 2 pieces: Normal 9&10) Small intestine, 4 pieces: Mucosa--moderately severe, diffuse eosinophilia; focal coccidian parasite. 11) Abomasum, 2 pieces: Mucosa--mild, multifocal eosinophilia. 12) Omasum: Epithelium-abscesses, moderately severe, acute, multifocal. 13) Reticulum: Epithelium-abscesses, mild, acute, multifocal; tunica muscularis--granuloma, minimal, focal with intralesional plant foreign body. 14) Thymus: Normal Perry L. Habecker, VMD OOOXFTOr EID151748 RGS000951 NEW BOLTON CENTER University of Pennsylvania, New Bolton Center Laboratory of Large Animal Pathology and Toxicology 382 W. Street Road Kennet! Square, PA 19348 Phone: (610)444-5800 Fax: (610)925-8110 FINAL REPORT Accession No.: UP 9903263 Vet: Dr. Lisa Davis-Heller Submitter Dr. Perry Habecker Pg#: 1 Report Address: j ) r y R y j, investigation Case Tracking #: Toxicology Case Coordinator: Robert H. Poppenga, DVM, PhD Date Submitted: 7/29/99 Report Date: 8/19/99 TA(D): 21 Species: Bovine Production type: Beef Breed: Sex: F A ge:.... .7 y Animal I.D. #37 Sample . Frozen Tissue Date obtained: Reference Lab: m w d Adult O Fetus O Juvenile O Unknown HISTORY SUMMARY: Killed and necropsied for Dr Lisa Heller on June 10, 1999. Tissues also submitted to histology (see 9902707) DIAGNOSIS: Copper deficiency. COMMENTS: The following bovine liver mineral ranges can be utilized for the interpretation of the analytical results contained in this report The indicated ranges are only guidelines and the results need to be interpreted in conjunction with management dietary, and clinical variables. Herd evaluations should be based upon analysis of samples from an adequate number of animals. Mineral Calcium Cobalt Copper Iron Magnesium Manganese Molybdenum Selenium Zinc Deficient < 40 ppm < 0.005 ppm < 25 ppm < 30 ppm < 1.00 ppm 0 02 to 0.17 j m < 20 ppm Marginal 0.005 to 0.017 ppm 1.50 to 3 00 ppm 0.12 to 0.25 ppm 25 to 40 ppm Adequate 30 to 200 ppm 0.020 to 0.085 ppm 25 to 150 ppm 45 to 300 ppm 100 to 200 ppm 0.14 to 1.40 ppm 0.25 to 0.50 ppm 25 to 200 ppm Ranges obtained from Puls' Mineral Levels in Animal Health. LABORATORY FINDINGS: TOXICOLOGY RESULTS: ) Sample: Liver and kidney 1s Test Complete Metal Screen Liver Kidney 1 Arsenic < 0.50 < 0.50 k I i 000071 46 EID151749 R6S000952 NEW BOLTON CENTER University of Pennsylvania, New Bolton Center Laboratory of Large Animal Pathology and Toxicology 382 W. Street Road Kcnnett Square, PA 19348 Phone: (610) 444-5800 Fax: (610) 925-8110 FINAL REPORT Accession No.: UP 9903263 V et Dr. Lisa Davis-Heller Submitter: Dr. Perry Habecker Pg#: 2 Cadmium Chromium Mercury Lead Selenium Thallium Calcium Cobalt Copper Iron Magnesium Manganese Molybdenum Selenium Zinc <0.10 <0.20 <1.00 <0.50 < 1.00 <2.00 42.4 <0.10 7.71 67.0 178 4.97 1.09 0.236 39.4 0.555 <0.20 <1.00 <0.50 < 1.00 <2.00 70.9 <0.10 4.29 93.9 148 1.66 <0.30 19.0 Beryllium Nickel Vanadium < 0 02 <0.30 < 0 30 < 0 02 <0.30 < 0 30 All metal results are reported as ppm on a tissue wet weight basis. Beryllium, vanadium and nickel axe pending Sample: Liver and fat Test: Organic Chemical Screen by GC/MS No toxic compounds were detected in the submitted sample by our gas chromatography - mass spectroscopy organic chemical screen. Both conventional extraction and SPME were used on the liver and fat samples. Sample: Rib Bone Test: Fluoride Fluoride was detected in the submitted bone sample at 1009 ppm (expressed on a fat free, dry bone basis). Robert H. Poppenga, DVM, PhD 000072 L? EID151750 R G S000953 Tennant Farm Herd Health Investigation Cattle Team Report DrWiur^afet^Plan Hazard 1. Physical injury from cattle | The team will follow the direction of the 3 most experienced cattle handlers (Bob M., Lisa, and Pete). Mr. Tennant and his brother will help with the handling of the cattle (note: added during 4/7/99 safety meeting). Rectal palpations will be performed only by veterinarians. All members of the cattle team will be responsible for their personal safety, the safety of the team, and the safety of other personnel in the animal handling area. All persons in the animal-handling area will be expected to abide by these safe practices. Physical restraint devices (corral, headgate, halters) will be examined for adequacy before use. Chemical restraint (e.g., sedatives) will be used on individual cattle if needed. 2. Physical injury from devices (e.g., knives. All members of the cattle team are experienced in the handling of veterinary tools. Necropsies, phlebotomies, needles) ear tagging, and similar procedures will be conducted only by the veterinarians. Disposable sharps (e.g., blades, needles) will be placed in an approved sharps container and returned to New Bolton Center. ! 3. Biological safety ; The cattle team will be appropriately dressed (coveralls I and boots) when working with the livestock. Soapy water, scrub brush, and paper towels will be available. Rectal sleeves will be used for rectal palpations. Latex gloves will be used for necropsies and when otherwise considered appropriate. (For the biosafety of the cattle, "single use" collection devices will be used.) If clinical signs warrant, the cattle team may implement additional biological safety measures._______________ 4. Chemical safetv Formalin will be used as a preservative during the | collection of necropsy tissues. All personnel will avoid _______________ ___________ |___skin contact with formalin.________________________ Notes: This Safety Plan will be reviewed by all members o f the cattle team prior to their arrival at Mr. Tennant's Farm in the Dry Run area, (note: plan was reviewed and accepted during safety meetingon 4/7/98 at the Parkersburg Holiday Inn. Cattle team and 4 EPA/USFWS persons present) The cattle team consists o f six veterinarians: Drs. Lisa Davis-Heller, Perry Habecker, Peter Moisan, Bob Munson, Bob Poppenga. and Greg Sykes._____________________________________________________________ Appendix C: Dry Run Safety Plan 000073 10 EID151751 R G S000954  Tennant Farm Herd Health Investigation Cattle Team Report Photo Photograph Legend # 1-41 Cattle 1 -4 1 . respectively. Each photo number corresponds to the ear tag number (placed following examination) of the cow, bull, or steer and the animal number in Table 1 of this report. 42 Cow # 42: The individual cow which escaped the run and was therefore not ear tagged or examined. 43 Livestock in corral. The 42 adult cattle (39 cows, 2 bulls, 1 steer) and 12 calves were held in this corral for several hours prior to examination in the late afternoon of April 7, 1999. 44 Cow # 25: Corneal scar 45 Cow # 26: Corneal opacity 46 Cow # 30: Corneal opacity 47 Cow # 37: Corneal scar 48 Cow # 40: Corneal opacity 49 Cow # 22: Mass (12 cm diameter), prescapular, subcutaneous, neck, right. 50 Cow # 22: Scalpel puncture and drainage of mass during examination. Brown-red fluid contents drained. 51 Cow # 22: Manual exploration of cystic cavity revealed a large mass of red hair within the cyst. Diagnosis was "epidermal inclusion cyst" of spontaneous origin which was of no significance to herd health. 52 Cow #31: Tongue with spotty melanosis. This was an incidental finding of no pathological significance. Melanosis of epidermal tissues, as seen here, is normal. 53 Cow # 18: Mammary gland lump adjacent to teat. This was considered to most likely be an abscess. 54 Group of cattle, owned by a neighbor of Mr. Tennant, grazing on an adjacent property. 55 Deer carcass on Tennant property. One of three deer carcasses observed by Mr. Tennant and Dr. Sykes, within 500 feet of the Tennant barn on April 8, 1999. All three appeared to have died in the past few' months. 56 Tennant barn and barnyard adjacent to the corral where the cattle examinations were conducted. 57 Grazing pasture along the Dry Run Creek between the landfill site and the Tennant barn. 58 Grazing pasture along the Dry Run Creek between the landfill site and the # Tennant barn. 9 59 Group of 16 cow skulls (photo supplied by Mr. Tennant) m 60 Worn incisors of an unidentified cow. bull, or steer (photo supplied by Mr. Tennant). ______ _____________________________ m 9 * 9 9 9 9 Appendix D: Phoios 1-66 9 9 7/ 9, 000074 RGS000955 Tennant Farm Herd Health Investigation Cattle Team Report Photo # 61 62 63 64 65 66 Photograph Legend (continued) Neck alopecia in an unidentified animal (photo supplied by Mr. Tennant). The cattle team considered this most likely due to lice. Neck alopecia in an unidentified animal (photo supplied by Mr. Tennant). The cattle team considered this most likely due to lice. Nose with spotty melanosis (photo supplied by Mr. Tennant). The cattle team considered this pigmentation to be normal. Nose with diffuse melanosis (photo supplied by Mr. Tennant). The cattle team considered this pigmentation to be normal. Hard palate with spotty melanosis (photo supplied by Mr. Tennant). The cattle team considered this pigmentation to be normal. Cattle team (CT) and steering committee (SC) members at the Tennant Farm during the April 7-8 1999 site visit. From left to right: Dr. Robert Poppenga (CT: veterinary toxicologist), Dr. Robert Munson (CT: veterinary clinician): Dr. Peter Moisan (CT: veterinary clinician/pathologist), Sarah Caspar (SC: EPA On-Scene Coordinator), Dr. Perry Habecker (CT: veterinary pathologist), Dr. Lisa Davis-Heller (CT: veterinary clinician). Dr. Michael Home (SC: USFWS, Environmental Response Team). Not shown: Dr. Greg Sykes (CT: veterinary pathologist/toxicologist; taking picture); Dr. Ralph Stahl (SC: DuPont biologist; not present) and Dr. Rudolph Valentine (SC: DuPont toxicologist; not present). Appendix D: Photos 1-66 000075 72- EID151753 R G S000956 i * 4( , i ,i r ,s ' 1* s* > * ' ` 4 1 I ' i f t ,V* '* }9 6 8 , - ^ |t- <-*s' I ' >* 1'M l '" ' ' RGS009 m -'000096 E ID l51774 R G S000977 Vf If I 1 I c l, EID1517E3 R , > .mW Tennant Farm Herd Health Investigation Cattle Team Report Herd Investigation - Beef Cattle - Date: April 8. 1999_______ Name: Wilbur Earl Tennant Address: Route 3 Box 17. Washington. WV 26181_________ [Derived from interview of Mr. Earl Tennant by Drs. Peter Moisan, Lisa Heller, and Robert Munson on 4/8/99] PARTI: 1998 A. History from last calvins season Months during which cows calved: All year, 1998. Number of cows in herd, 1998 calving season: 40 . Number of calves born: 35 live. Number of cows calved 35_ or 87.5 % of cows exposed. Abortions/Stillborn in 1998: 5 - 6 Other calving problems in 1998: None . Sick calves in 1998: Scours: About 6 (No deaths') . Weak calves in 1998: Several were weak and had enlarged joints. Other illness in unweaned calves in 1998: Most calves were slow-growing and underweight during nursing period. Birth weight was low in the calves born in 1998 . Age at weaning in 1998: 5 m os.. Number of calves weaned: 30 or % of cows exposed: 75%. Illness in weaned calves in 1998: All calves were sold at weaning time. A few calves (5). born in 1998 (fall) are still nursing the cows. B. Cows Illness in cows in 1998: 1 cow disappeared in 1998. Ocular disease: Some cattle have darker eves when put in the hollow during the summer. The darker eves resolve when they are kept away from the creek. Skin disease: Some with lesions around the coronary bands. Thought to be "mild". Diarrhea: A few cows had diarrhea when calves were 3 - 4 weeks old. Lameness: Cows seem lame in cold damp weather (arthritis?). Respiratory disease: 3 cows had a transient epistaxis in 1998. Other disease in 1998: Cows and calves seem more healthy than in earlier years. Treatments for disease in 1998: Alum to cows for calf scours. Treated cows with ground garlic for worms. Vaccinations in herd in 1998: None. Brucellosis vaccinations are not done in WV. Diagnostic work performed in herd in 1998: Include necropsy or blood sampling: None. (4 years ago. 12 blood samples were taken and paired serology was inconclusive for brucellosis and leptospirosis). Estimated body condition score (BCS range 1-9) of cows in 1998 calving season: Same as 1999. May have been slightly better BCS than 1999. Appendix E: Herd Health History /O. ^00109 EID151787 R6S000990 Tennant Farm Herd Health Investigation C. Feeding History Cattle Team Report Feed to cows during calving season 1998 (Include minerals): Com silage. Ground corn with supplement. Orchard grass/timothv/brome/KY31 fescue hav. Fairly high percentage fescue. 5 pounds of grain per cow per day, with slightly more during cold weather. Feed to cows in winter before calving season in 1998 (Include minerals): Same as during calving season with higher mineral. 5 pounds of grain per cow per dav. with slightly more during cold weather. Feed to nursing cows during grazing season 1998 (Include minerals): Trace mineral salt (TDN salt). Feed Test Results for 1998: None. D. Other Other comments for 1998: Since the problems first started in 1988. the acreage grazed has decreased from 514 acres in 1988 to 196 acres now. In 1988 there were 200 cows. Silage was fed to these cows in the winter. Since 1988 there have been 176 dead cows and about 200 calves. The first to die appeared to have pneumonia, with shivering, fever, and foaming from the mouth. Other cows (15-20) were circling, then died.__________________ PART II: 1999 A. History from current calving season First calving: About March 1. 1999 . Length of breeding season 1998: year long. 1998. Anticipated last calving date in 1999: About October 1999 or later . Number of cows in herd. 1999 calving season: 39.. Number of calves born so far: 7_. Number of cows calved so far: 7. or 18 % of cows exposed. Abonions/stillbirths so far in 1999: 1 Other calving problems in 1999: i stillborn in 1999 and 1 placenta found without a dead cal f. Sick calves in 1999: Scours None . Weak calves in 1999: None . Other illness in unweaned calves in 1999: A few of last fall's calves, still nursing cows on pasture: are loose. Birth weights are around 35 pounds and appear lower than normal. Calves are fairly thin but otherwise healthy in 1999. B. Cows Illness in cows in 1999: None . Ocular disease:____________ . Skin disease: ____________ . Diarrhea: . Lameness: . Appendix E: Herd Healih History l'7 O O O llO EID151788 RGS000991 Tennant Farm Herd Health Investigation Cattle Team Report Respiratory disease: None . Other disease in 1999: Cows are healthy in 1999, so far. Owner notices that the cows are fairly thin . Treatments for disease in 1999: None . Vaccinations in herd in 1999: None . Diagnostic work performed in herd in 1999: Include necropsy or blood sampling: None . Owner estimated body condition score (BCS-range 1-9) of cows in 1998 breeding season: Clinical BCS of cows (average) on day of herd visit: 3 - 3.5 (Range BCS 2 - 7 ) . C. Feeding History Feed to cows during calving season 1999 (Include minerals): Hav and ground feed as in 1998. No silaee . Feed to cows in winter before calving season 1999 (Include minerals): Ground feed, same as in 1998, with higher mineral mix . Feed to cows during grazing season 1999 (Include minerals):_____ . Feed Test Results for 1999: None . D. Other Other comments for 1999 to date: It seems that problems with cows and calves have decreased by 75 - 80% since the water from the landfill was hauled awav for treatment. No new cows have been purchased in past 5 years. Bulls were purchased in 1996. estimated, but could have been in 1995. Bulls haven't been sick or poor breeders. There are reports of 2 neighbors with premature calves born, for total of 6 calves . Appendix E: Herd Health History OOOJ fO & EID1517B9 R G S000992 a Results for: Tennant Lactating beef Mooeu 1 Ration Nutrients SuDDlied and Reauired ME Avail ME Reqd Mcal/d Mcal/d Totals 16 23 Percent difference Maintenance 16 15.1 Pregnancy 10 Lactation 17 Gain Reserves -6 0 06 Difference Mcal/d -6 72.5% 1 1 -6 -6 -6 MP Avail g/d 639 639 320 320 -43 0 Intake and Performance Predictions DMI predicted DMI entered-% predicted: 66% Relative DMI Target ADG w/conceptus ME Allowable Gain MP Allowable Gain AA Allowable Gain 23.3 Ibs/d 15.3 Ibs/d 100% 0.24 Ibs/d 0.11 Ibs/d -0.17 Ibs/d 0.02 Ibs/d Pred. Max Forage Intake Entered Forage Intake ME Allowable Milk MP Allowable Milk AA Allowable Milk Days to lose 1 C S 12/15/99 MP Reqd Difference g/d g/d 702 -64 90.9% 318 320 0 320 363 -43 21 -64 64 -64 12.0 Ibs/d 12.0 Ibs/d 13.7 Ibs/d 11.3 lbs/d 13.9 Ibs/d 43 Diet Concentrations and Rumen Balances Effective NDF required 3.1 Ibs/d Effective NDF supplied 8.7 Ibs/d NDF in ration 61% DM Diet ME 1.07 MCAL/LB DM Diet NEI 0.69 MCAL/LB DM Diet NEm 0.67 MCAL/LB DM Diet NEg 0.41 MCAL/LB DM Pred. Ruminal pH 6.46 Q/d % of reauirements Ruminal N Balance -26 79% Pept Bal. 22 177% First limiting AA: MET 1.19 109% 2nd limiting AA: LY S 2.29 106% MP from bacteria MP from undeg feed Diet CP DIP Soluble Protein Total N FC in ration Fat in ration (total): Total N balance Predicted MUN Urea cost % Forage in ration Ration DM DMI/Maint DMI Predicted Excretion N excretion: Fecal Urinary Total P excretion: Fecal Urinary Total Ration Costs Cost/day Cost/cwt. milk production App*\*tJ\.y F 0.17 $/day 1.27 $/cwt Cost/cwt. ME allow, milk Cost/cwt. MP allow, milk Cost/cwt. AA allow, milk 467 g/d 172 g/d 8.6% DM 61% CP 23% CP 18% DM 4.7% DM 0 g/d 0.00 Mcal/d 71% DM 90% 1.1 X Maint. 0.2 lbs/hd/d 0.1 lbs/hd/d 0.3 lbs/hd/d 37.2 g/hd/d 0.9 g/hd/d 38.1 g/hd/d 1.27 $/cwt 1.54 $/cwt 1.25 $/cwt /*? OOOUU2 EID151790 R G S000993 I Ration for Tennant Lactating beef cow Enter amt. fed in blue cells: As-fed Lbs Forage Pasture Rel DMI * Pounds % of ration Home grown As-fed as fed As-fed Fescue, K31 Corn Dry Soybean Hay, F. bloom Ear45 Meal 44 12.00 4.20 0.30 y n n n n n y 12.00 12.00 70.6% y 4.20 4.20 24.7% n 0.30 .30 1.8% Blank 0.00 n n n 0.00 .00 0.0% Blank 0.00 n n n 0.00 .00 0.0% Blank 0.00 n n n 0.00 .00 0.0% Blank 0.00 n n n 0.00 .00 0.0% Blank 0.00 n n n 0.00 .00 0.0% Blank 0.00 n n n 0.00 .00 0.0% Blank 0.00 n n n 0.00 .00 0.0% Blank 0.00 n n n 0.00 .00 0.0% Dry Cow Min 0.50 n n n 0.50 .50 2.9% Blank 0.00 n n n 0.00 .00 0.0% Blank 0.00 n n n 0.00 .00 0.0% Total 17.00 17.00 17.00 100% DMI Predicted 23.28 Entered DM - Predicted -7.98 DMI Entered: 15.30 Relative DMI % 100.0% MP Balance -63.8 g/d 91% Pred DMI 23.3 IbS/d ME Balance mcal/d 73% Actual DMI 15.3 Ibs/d Rumen N Balance -26.5 g/d -21% Effective NDF Balance 5.6 Ibs/d Peptide Balance 21.7 g/d 77% Days to lose 1 C S 43 O Cost : $./day $ 0.17 J* t CM CD EID151791 0w <5 Page 1 General Factors Farm Name: Diet Number in group: Days to feed NDF capacity Units Feed Entry Basis LB/Head/Day Milk Price Feed Losses File Name Animal Factors Grade Animal Type Age Sex Body Weight Breed Type Mature Weight Condition Score Breeding System Dam breed Dam's maternal Dam's paternal Days Pregnant Days Since Calving Lactation # Rolling Herd Average (dairy) Milk Production (dairy) Milk Fat (dairy) Milk Protein (dairy) assume C P Relative Milk Prod'n (beef) Expected Calf Birth Weight Management Factors Additive Grazing Unit Size Daily pasture allowance initial pasture mass Selection-Pressure for growth/milk Feeding Frequency Feeding Method Calf Implanted Environmental Factors Wind Speed Previous Temperature Previous RH Current Temperature Tennant Lactating beef cow 1 365 1 % of BW 1 English 1 As-Fed /cwt 0% % dry matter :\casestudy\test 5 5 Enter 1 to F A L S E 0 Lactating beef cow (predicted milk 70 Months 4 Cow 900 Lb 1 Beef Maturity 900 Lb 3 1=v.thin - 9=v.fleshy 2 2way cross 17 #N/A Hereford 1 1 Angus 17 17 Hereford 18 18 0 Days 30 Days DLW 4 0=dry or heifer 0 Lb Predicted values 0.00 Lb 7.0 0.00 % 3.6 0% 3.8 2 1-9 16.53 45 Lb 73.8 0 1 none 0.00 Acres 0 DM Ib/PDMI 1847 lb DM/acre 0 1-5 Scale 2 # of Times Fed Daily 2 1=forage&grain separate,2=TMR 1 1=no,2=yes 0 5 mph 40 Degrees F 30 % 40 Degrees F Page 1 12/15/99 3:28 PM /// 00013.4 EID151792 RGS000995 Current RH Storm Exposure Night Cooling Hair Depth Hide Hair Coat Cattle Panting (Heat Stress) DM1 Scaler In summer, cattle are exposed to: Rectal Temperature (optional) Animal Activity Functions Time spent standing Number of body position changes Distance walked Flat Slooed Mud Depth Feedbunk Characteristics Bunk level Bunk Surface Target Growth Age at tst calving Herd Calving Interval 30 % 1 1=no,2=yes 1 1=none,2=with night cooling 1 in 2 1=thin,2=avg,3=thick 1 1=no mud,2=some mud on lower b 1 1=no,2=rapid shallow,3=open mout 1 % (110=+10%; 90= -10%) 12 1=No direct sunlight, 2=direct sunlig 101.5 Degrees F 0 18 hrs per day 6 (lying down and standing again) 6562 feet/d feet/d 0 inches 6 inches (distance between cow floor 1 see codes-----> 24 months 16 months Page 2 000115 12/15/99 3:28 PM tt? - EID151793 RGS000996 Results for: Tennant Lactating beef MoO EL X Ration Nutrients Supplied and Required ME Avail ME Reqd Mcal/d Mcal/d Totals 23 23 Percent difference Maintenance 23 15.0 Pregnancy 80 Lactation 87 Gain 0 0 Reserves 00 Difference Mcal/d 0 100.2% 8 8 0 0 0 MP Avail g/d 883 883 508 507 144 123 Intake and Performance Predictions DMI predicted DMI entered-% predicted: 85% Relative DMI Target ADG w/conceptus ME Allowable Gain MP Allowable Gain AA Allowable Gain 23.9 Ibs/d 20.3 Ibs/d 100% 1.26 Ibs/d 0.11 Ibs/d 0.65 Ibs/d 0.02 Ibs/d Pred. Max Forage Intake Entered Forage Intake ME Allowable Milk MP Allowable Milk AA Allowable Milk Days to gain 1 C S 12/15/99 MP Reqd Difference g/d g/d 759 123 116.3% 375 508 0 507 363 144 21 123 0 123 12.0 Ibs/d 12.0 Ibs/d 13.7 Ibs/d 18.3 Ibs/d 22.8 Ibs/d 7586 Diet Concentrations and Rumen Balances Effective NDF required 4.1 Ibs/d Effective NDF supplied 9.5 Ibs/d NDF in ration 54% DM Diet ME 1.11 M C A L ` 3 DM Diet NEI 0.72 MCAL/LB DM Diet NEm 0.71 MCAL/LB DM Diet NEg 0.44 MCAL/LB DM Pred. Ruminal pH 6.46 Ruminal N Balance Pept Bal. m % of requirements -40 76% 13 124% First limiting AA: MET 5.45 139% 2nd limiting AA; L Y S 13.80 131% MP from bacteria MP from undeg feed Diet C P DIP Soluble Protein Total N FC In ration Fat in ration (total): Total N balance Predicted MUN Urea cost % Forage in ration Ration DM DMI/Maint DMI Predicted Excretion N excretion: Fecal Urinary Total P excretion: Fecal Urinary Total Ration Costs Cost/day Cost/cwt. milk production 0.17 $/day 1.27 $/cwt Cost/cwt. ME allow, milk Cost/cwt. MP allow, milk Cost/cwt. AA allow, milk 636 g/d 246 g/d 8.7% DM 60% CP 21% CP 28% DM 4.4% DM 20 g/d 0 22 Mcal/d 53% DM 69% 1.5 X Maint. 0.2 lbs/hd/d 0.1 lbs/hd/d 0.4 lbs/hd/d 43.2 g/hd/d 0.9 g/hd/d 44.2 g/hd/d 1.27 $/cwt 0.95 $/cwt 0.76 $/cwt U3 000116 .EID151794. R G S000997 Ration for Tennant Lactating beef cow Enter amt. fed in blue cells: As-fed Lbs Forage Pasture Rei DMI * Pounds % of ration Home grown As-fed as fed As-fed Fescue, K31 Corn Dry Soybean Hay, F. bloom Ear45 Meal - 44 12.00 10.00 0.30 Blank 0.00 Blank 0.00 Blank 0.00 Blank 0.00 Blank 0.00 Blank 0.00 Blank 0.00 Blank 0.00 Dry Cow Min Blank Blank 0.50 0.00 0.00 Total 22.80 DMI Predicted 23.86 Entered DM - Predicted Relative DMI % -3.57 100.0% MP Balance ME Balance 123.5 g/d 0.0 mcal/d Rumen N Balance 39.9 g/d Peptide Balance 12.6 g/d yn nn nn nn nn nn nn nn nn nn nn nn nn nn y y n n n n n n n n n n n n DMI Entered: 116% Pred DMI 100% Actual DMI -24% Effective NDF Balance 24% Days to gain 1 C S 12.00 10.00 0.30 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.50 0.00 0.00 22.80 12.00 10.00 .30 .00 .00 .00 .00 .00 .00 .00 .00 .50 .00 .00 22.80 20.29 23.9 Ibs/d 20.3 Ibs/d 5.5 Ibs/d 7586 52.6% 43.9% 1.3% 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 2.2% 0.0% 0.0% 100% Cost : $./day $ 0.17 EID151795 WQ Page 1 Results for: Tennant Dry pregnant m o del 3 Ration Nutrients Suoplied and Required ME Avail ME Reqd Mcal/d Mcal/d Totals 16 22 Percent difference "Maintenance 16 15.3 Pregnancy 13 Lactation -1 0 Gain -1 4 Reserves 05 Difference Mcal/d -5 75.2% 1 -1 -1 -5 -5 12/15/99 U5/W6 fcftbC e pfitEOlCTtoM E'Q C A rro ld MP Avail g/d 639 639 320 235 235 113 MP Reqd Difference g/d g/d 526 113 121.4% 318 320 85 235 0 235 122 113 0 113 Intake and Performance Predictions DMI predicted DMI entered-% predicted: 83% Relative DMI Target ADG w/conceptus ME Allowable Gain MP Allowable Gain AA Allowable Gain ADGpreg Conceptus Weight & Tissue 18.5 Ibs/d 15.3 Ibs/d 100% 0.24 Ibs/d 1.23 Ibs/d 1.82 Ibs/d 0.58 Ibs/d 0.58 Ibs/day 43.86 lbs DMI predicted (Chase) Pred. Max Forage Intake Entered Forage Intake ME Allowable Milk MP Allowable Milk AA Allowable Milk Days to lose 1 C S 12.0 Ibs/d 12.0 Ibs/d 0.0 Ibs/d 0.0 Ibs/d 0.0 Ibs/d 49 Diet Concentrations and Rumen Balances Effective NDF required 3.1 Ibs/d Effective NDF supplied 8.7 Ibs/d NDF in ration 61% DM Diet ME 1.07 MCAL/LB DM Diet NEI 0.69 MCAL/LB DM Diet NEm 0.67 MCAL/LB DM Diet NEg 0.41 MCAL/LB DM Pred. Ruminal pH 6.46 a/d % of reauirements Ruminal N Balance -26 79% Pept Bal. 22 177% First limiting AA: MET 4.59 149% 2nd limiting AA: L Y S 12.59 142% MP from bacteria MP from undeg feed Diet C P DIP Soluble Protein Total NFC In ration Fat in ration (total): Total N balance Predicted MUN Urea cost % Forage in ration Ration DM DMI/Maint DMI 467 g/d 172 g/d 8.6% DM 61% CP 23% CP 18% DM 4.7% DM 18 g/d 0.20 Mcal/d 71% DM 90% 1.1 X Maint. Predicted Excretion N excretion: Fecal Urinary Total P excretion: Fecal Urinary Total 0.2 lbs/hd/d 0.1 lbs/hd/d 0.3 lbs/hd/d 30.3 g/hd/d 0.8 g/hd/d 31.1 g/hd/d Ration Costs Cost/day Cost/cwt. milk production fip p e ijji > 0.17 5/day #N/A $/cwt Cost/cwt. ME allow, milk Cost/cwt. MP allow, milk Cost/cwt. AA allow, milk #N/A #N/A #N/A $/cwt $/cwt $/cwt ns 000118 EID151796 R G S 0 0 0 9 9 8 .0 1 Ration for Tennant Dry pregnant cow Enter amt. fed in blue cells: As-fed Lbs Forage Pasture Rel DMI * Pounds % of ration Home grown As-fed as fed As-fed Fescue, K31 Corn Dry Hay, F. bloom Ear45 12.00 4.20 Soybean Meal - 44 0.30 Blank 0.00 Blank 0.00 Blank 0.00 Blank 0.00 Blank 0.00 Blank 0.00 Blank 0.00 Blank Dry Cow Min 0.00 0.50 Blank Blank 0.00 0.00 Total 17.00 DMI Predicted 18.46 Entered DM - Predicted -3.16 Relative DMI % I 100.0% MP Balance ME Balance 112.7 g/d -5.4 mcal/d Rumen N Balance Peptide Balance -26.5 g/d 21.7 g/d yn nn nn nn nn nn nn nn nn nn nn nn nn nn y y n n n n n n n n n n n n DMI Entered: 121% Pred DMI 75% Actual DMI -21% Effective NDF Balance 77% Days to lose 1 C S 12.00 4.20 0.30 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.50 0.00 0.00 17.00 12.00 4.20 .30 .00 .00 .00 .00 .00 .00 .00 .00 .50 .00 .00 17.00 15.30 18.5 Ibs/d 15.3 Ibs/d 5.6 Ibs/d 49 70.6% 24.7% 1.8% 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 0.0% 2.9% 0.0% 0.0% 100% Cost : $./day $ 0.17 W Owooo m 5 10 01 to to <-40 Page 1 Farm Name: Diet Number in group: Days to feed NDF capacity Units Feed Entry Basis LB/Head/Day Milk Price Feed Losses File Name Animal Factors Grade Animal Type Age Sex Body Weight Breed Type Mature Weight Condition Score Breeding System Dam breed Dam's maternal Dam's paternal Days Pregnant Days Since Calving Lactation # Rolling Herd Average (dairy) Milk Production (dairy) Milk Fat (dairy) Milk Protein (dairy) assume C P Relative Milk Prod'n (beef) Expected Calf Birth Weight Management Factors Additive Grazing Unit Size Daily pasture allowance Initial pasture mass Selection-Pressure for growth/milk Feeding Frequency Feeding Method Calf Implanted Environmental Factors Wind Speed Previous Temperature Previous RH Current Temperature Tennant Dry pregnant cow U 5 / iV ^ m / ? eu >r i 1 F o q c a P f t O lC .Tlc"ts 365 1 % of BW E Q u TIC fV 1 English 1 As-Fed $ /cwt 0% % dry matter C:\casestudy\test 5 5 Enter 1 to FA LSE 3 Dry Cow 70 Months 4 Cow 900 Lb 1 Beef Maturity 900 Lb 3 1=v.thin - 9:=v. fleshy 2 2way cross 17 #N/A Hereford 1 1 Angus 17 17 Hereford 18 18 240 Days 360 Days DLW 4 0=dry or heifer 0 Lb Predicted vai 45.40 Lb 1.2 3.70 % 6.8 3% 4.5 1 1-9 16.53 45 Lb 73.8 0 1 none 0.00 Acres 0 DM Ib/PDMI 1847 lb DM/acre 0 1-5 Scale 2 # of Times Fed Daily 2 1=forage&grain separate,2=TMR 1 1=no.2=yes 0 5 mph 40 Degrees F 30 % 40 Degrees F Page 1 12/15/99 1:25 PM m ______________________________ EID151798 000120 R G S O O IO O O Current RH Storm Exposure Night Cooling Hair Depth Hide Hair Coat Cattle Panting (Heat Stress) DMI Scaler In summer, cattle are exposed to: Rectal Temperature (optional) Animal Activity Functions Time spent standing Number of body position changes Distance walked Flat Sloped Mud Depth Feedbunk Characteristics Bunk level Bunk Surface Target Growth Age at 1st calving Herd Calving Interval 30 % 1 1=no,2=yes 1 1=none,2=with night cooling 1 in 2 1=thin,2=avg,3=thick 1 1=no mud,2=some mud on lower b 1 1=no,2=rapid shallow,3=open mout 1 % (110=+10%; 90= -10%) 12 1=No direct sunlight, 2=direct sunlig 101.5 Degrees F 0 18 hrsperday 6 (lying down and standing again) 6562 feet/d feet/d 0 inches 6 inches (distance between cow floor 1 see codes-----> 24 months 16 months Page 2 000121 12/15/99 1:16 PM ns EID151799 R G S O O lO O l Results for: Tennant Dry pregnant 12/15/99 r A o p e t , H- UIW C IN TA KE" (O f* * ) /VlATT-<r<3. Ration Nutrients SuDDlied and Required ME Avail ME Reqd Mcal/d Mcal/d Totals 19 22 Percent difference Maintenance 19 15.5 Pregnancy 43 Lactation 10 Gain 1 4 Reserves 03 Difference MP Avail Mcal/d ____________________ ________ -3 762 87.7% 4 762 1 385 1 300 3 300 3 177 MP Reqd Difference g/d g/d 584 177 130.3% 377 385 85 300 0 300 122 177 0 177 Intake and Performance Predictions DMI predicted DMI entered-% predicted: 100% Relative DMI Target ADG w/conceptus ME Allowable Gain MP Allowable Gain AA Allowable Gain ADGpreg Conceptus Weight & Tissue 18.5 Ibs/d 18.5 Ibs/d 100% 0.68 Ibs/d 1.23 Ibs/d 2.17 Ibs/d 0.58 Ibs/d 0.58 Ibs/day 43.86 lbs DMI predicted (Chase) Pred. Max Forage Intake Entered Forage Intake ME Allowable Milk MP Allowable Milk AA Allowable Milk Days to lose 1 C S 12.0 Ibs/d 15.5 Ibs/d 0.0 Ibs/d 0.0 Ibs/d 0.0 Ibs/d 97 Diet Concentrations and Rumen Balances Effective NDF required 3.7 Ibs/d Effective NDF supplied 11.0 Ibs/d NDF in ration 63% DM Diet ME 1.05 MCAL/LB DM Diet NEI 0.68 MCAL/LB DM Diet NEm 0.65 MCAL/LB DM Diet NEg 0.39 MCAL/LB DM Pred. Ruminal pH 6.46 a/d % of reauirements Ruminal N Balance -31 79% Pept Bal. 27 190% First limiting AA: MET 5.90 156% 2nd limiting AA: L Y S 16.66 149% MP from bacteria MP from undeg feed Diet C P DIP Soluble Protein Total N FC in ration Fat in ration (total): Total N balance Predicted MUN Urea cost % Forage in ration Ration DM DMl/Maint DMI Predicted Excretion N excretion: Fecal Urinary Total P excretion: Fecal Urinary Total Ration Costs Cost/day Cost/cwt. milk production App*SSc/>y. P 0.17 $/day #N/A $/cwt Cost/cwt. ME allow, milk Cost/cwt. MP allow, milk Cost/cwt. AA allow, milk 548 g/d 214 g/d 8.4% DM j 0% CP 23% CP 16% DM 4.8% DM 28 g/d 0.32 Mcal/d 76% DM 90% 1.3 X Maint. 0.2 lbs/hd/d 0.1 lbs/hd/d 0.3 lbs/hd/d 32.2 g/hd/d 0.8 g/hd/d 33.0 g/hd/d #N/A #N/A #N/A $/cwt $/cwt S/cwt II? 000122 EID151800 R G S001002 Ration for Tennant Dry pregnant cow Enter amt. fed in blue cells; As-fed Lbs Forage Pasture Rei DMI * Pounds % of ration Home grown As-fed as fed As-fed Fescue, K31 Corn Dry Soybean Blank Hay, F, bloom Ear45 Meal - 44 15.50 4.20 0.30 0.00 y n n n n n n n y 15.50 15.50 75.6% y 4.20 4.20 20.5% n 0.30 .30 1.5% n 0.00 .00 0.0% Blank Blank Blank Blank Blank 0.00 n n n 0.00 .00 0.0% 0.00 n n n 0.00 .00 0.0% 0.00 n n n 0.00 .00 0.0% 0.00 n n n 0.00 .00 0.0% 0.00 n n n 0.00 .00 0.0% Blank 0.00 n n n 0.00 .00 0.0% Blank Dry Cow Min 0.00 n n n 0.00 .00 0.0% 0.50 n n n 0.50 .50 2.4% Blank 0.00 n n n 0.00 .00 0.0% Blank 0.00 n n n 0.00 .00 0.0% Total 20.50 20.50 20.50 100% DMI Predicted 18.46 Entered DM - Predicted Relative DMI % MP Balance ME Balance 0.02 | 100.0% 177.2 g/d -2.7 mcal/d DMI Entered: 130% Pred DMI 88% Actual DMI 18.49 18.5 Ibs/d 18.5 Ibs/d O Rumen N Balance Peptide Balance O 1-* 10 CO -30.5 g/d 27.2 g/d -21% Effective NDF Balance 90% Days to lose 1 C S Cost ; $./day 7.3 Ibs/d 97 $ 0.17 Q o CO Page 1  tft Tennant Farm Herd Health Inveslijation Cattle Team Report (blank page) 000124 EID151802i RGS 0 0 1 0 0 4 NEW BOLTON CENTER University of Pennsylvania, New Bolton Center Laboratory of Large Animal Pathology and Toxicology 382 W. Street Road Kennett Square, PA 19348 Phone: (610) 444-5800 Fax: (610) 925-8110 FINAL REPORT Accession No.: UP 9902707 Submitter Dr. Perry Habecker Pg#: 1 Report Address: Dry Run Investigation Team Case Tracking #: N o.p. Case Coordinator: Perry L. Habecker, VMD Date Submitted: 6/27/99 Report Date: 7/5/99 TA(D): 8 Species: Bovine Production type: Breed: Sex: F Age:..... .7.y Animal I.D. #37 Sample -- 14--. Fixed Tissue Date obtained: Reference Lab: m w d Adult O Fetus O Juvenile O Unknown HISTORY SUMMARY: ~'illed and necropsied for Dr. Lisa Heller on June 10, 1999. Tissues also submitted to toxicology. DIAGNOSIS: Enteric lesions o f minimal significance. COMMENTS: The gastrointestinal eosinophilia is attributed to previous and current bouts of endoparasitism. Epithelial abscessation in the forestomachs is associated with acidosis due to excessive carbohydrate ingestion. LABORATORY FINDINGS: HISTOPATHOLOGY EXAMINATION: 1) Heart, 2 pieces: Minimal, focal lymphocytic myocarditis; focal Sarcocyst spp.. 2) Lung, 2 pieces: Normal 3) Liver, 2 pieces Normal 4) Spleen: Normal 5) Kidney: Normal 6) T eat Normal 7) Adrenal, 2 pieces: Normal 8) Lymph node, 2 pieces: Normal 9&10) Small intestine, 4 pieces: Mucosa--moderately severe, diffuse eosinophilia; focal coccidian parasite. 11) Abomasum, 2 pieces: Mucosa--mild, multifocal eosinophilia. 12) Omasum: Epithelium--abscesses, moderately severe, acute, multifocal. 13) Reticulum: Epithelium--abscesses, mild, acute, multifocal; tunica muscularis--granuloma, minimal, focal with intralesional plant reign body. .) Thymus: Normal Perry L. Habecker, VMD 000125 DUP 004 Photo Photograph Legend ____# ________________________________________________________________________________________ _____ 1-41 Cattle 1-41, respectively. Each photo number corresponds to the ear tag number (placed following examination) of the cow, bull, or steer and the animal number in Table 1of this report. 42 Cow # 42: The individual cow which escaped the run and was therefore not ear tagged or examined. 43 Livestock in corral. The 42 adult cattle (39 cows, 2 bulls, 1steer) and 12 calves were held in this corral for several hours prior to examination in the late afternoon of April 7, 1999. 44 Cow # 25: Comeal scar 45 Cow # 26: Comeal opacity 46 Cow # 30: Comeal opacity 47 Cow # 37: Comeal scar 48 Cow # 40: Comeal opacity 49 Cow # 22: Mass (12 cm diameter), prescapular, subcutaneous, neck, right. 50 Cow # 22: Scalpel puncture and drainage of mass during examination. Brown-red fluid contents drained. 51 Cow # 22: Manual exploration of cystic cavity revealed a large mass of red hair within the cyst. Diagnosis was "epidermal inclusion cyst" of spontaneous origin which was of no significance to herd health. 52 Cow #31: Tongue with spotty melanosis. This was an incidental finding of no pathological significance. Melanosis of epidermal tissues, as seen here, is normal. 53 Cow # 18: Mammary gland lump adjacent to teat. This was considered to most likely be an abscess. 54 Group of cattle, owned by a neighbor of Mr. Tennant, grazing on an adjacent property. 55 Deer carcass on Tennant property. One of three deer carcasses observed by Mr. Tennant and Dr. Sykes, within 500 feet of the Tennant bam on April 8,1999. All three appeared to have died in the past few months. 56 Tennant barn and barnyard adjacent to the corral where the cattle examinations were conducted. 57 Grazing pasture along the Dry Run Creekbetween the landfill site and the Tennant bam. 58 Grazing pasture along the Dry Run Creek between the landfill site and the Tennant bam. 59 Group of 16 cow skulls (photo supplied by Mr. Tennant) 60 Worn incisors of an unidentified cow, bull, or steer (photo supplied by Mr. ______________Tennant)._______________ ______________________________ DRAFT 07/30/99 000126 DUP 028 Photo # 61 62 63 64 65 66 Photograph Legend (continued) Neck alopecia in an unidentified animal (photo supplied by Mr. Tennant). The cattle teamconsidered this most likely due to lice. Neck alopecia in an unidentified animal (photo supplied by Mr. Tennant). The cattle teamconsidered this most likely due to lice. Nose with spotty melanosis (photo supplied by Mr. Tennant). The cattle teamconsidered this pigmentation to be normal. Nose with diffuse melanosis (photo supplied by Mr. Tennant). The cattle teamconsidered this pigmentation to be normal. Hard palate with spotty melanosis (photo supplied by Mr. Tennant). The cattle teamconsidered this pigmentation to be normal. Cattle team (CT) and steering committee (SC) members at the Tennant Farmduring the April 7-8 1999 site visit. From left to right: Dr. Robert Poppenga (CT: veterinary toxicologist), Dr. Robert Munson (CT: veterinary clinician); Dr. Peter Moisan (CT: veterinary clinician/pathologist), Sarah Caspar (SC: EPA On-Scene Coordinator), Dr. Perry Habecker (CT: veterinary pathologist), Dr. Lisa Davis-Heller (CT: veterinary clinician), Dr. Michael Home (SC: USFWS, Environmental Response Team). Not shown: Dr. Greg Sykes (CT: veterinary pathologist/toxicologist; taking picture); Dr. Ralph Stahl (SC: DuPont biologist; not present) and Dr. Rudolph Valentine (SC: DuPont toxicologist; not present). DRAFT 07/30/99 000127 1 -,; : - , >1 DUP. 036 '' 1 ,} .'H I i . 11- Afilli iB St i h l 053' J i ` I ' i ' ' : '5 r'-pP JOIBil P PPB r Afszb - \h-l*h 000161 DRAFT REPORT DRY RUN CREEK WASHINGTON, WOOD COUNTY, WEST VIRGINIA NOVEMBER 1997 PREPARED BY: Mark D. Sprenger, Ph.D. Environmental Response Team AND Michael T. Horne, Ph.D. U.S. Fish & Wildlife Service/Environraental Response Team IN CONJUNCTION WITH: Mark Huston REAC/ERT Environmental Response Team Center Office of Emergency & Remedial Response 000162 USEPA 6861 0 0 0 - it < .\ USFW 0579 TABLE OF CONTENTS LIST OF TABLES....................................................................................................................................... .... LIST OF FIGURES ....................................... ........................................................................................... viii SECTION I. 1.0 2.0 3.0 TECHNICAL APPROACH. SUMMARY OF FIELD EFFORT RESULTS. AND PRELIMINARY RISK SCREEN.................................................................................................................. 1 INTRODUCTION.............................................................................................................. 1 1.1 Objective .............................................................................................................. 1 1.2 Site Background.................................................................................................... I METHODOLOGY .................................................................................................. 1 2.1 Soil Sampling ....................................................................................................... 1 2.2 Sediment Sampling................................................................................................ 2 2.3 Surface Water Sampling .......................................................................................... 2 2.4 Drinking Water Well Sampling............................................................. '. ............. 2 2.5 Biological Sampling.............................................................................................. 3 2.5.1 Small Mammal Study................................................................................ 3 2.5.2 Vegetation Sampling................................................................................ 3 2.5.3 Aquatic Macroinvenebrate Sampling......................................................... 4` 2.5.4 Fish Collection......................................................................................... 4 2.6 Toxicity Testing..................................................................................................... 4 2.6.1 Eiseniafoerida (Earthworm) Toxicity Tests............................................... 4 2.6.2 Hyalella azseca (Amphipod) Toxicity Tests................................................ 5 2.6.3 Pimephalespromelas (Minnow) Toxicity Tests ........................................ 5 2.7 Sampling Equipment Decontamination.................................................................... 5 2.8 Standard Operating Procedures.............................................................................. 5 2.8.1 Documentation......................................................................... '............... 5 2.8.2 Sample Packaging, Shipment, Storage, Preservation, and Handling .......... 5 2.8.3 Field Sampling and Analytical Techniques................................................ 5 2.8.4 Health and Safety..................................................................................... 6 RESULTS.................................................................................................................'. . . 1 6 3.1 Water, Soil, and SedimentAnalysis ......................................................................... 6 3.1.1 BNAs ............................................................................................. 6 C0 k-J A -- 000163 USEPA 6862 USFW 0580 3.1.2 TAL Metals............................................................................................. 7 3.1.3 Pesticides/PCBs...................................................................................... 8 3.1.4 VOCs ................................................................................................... 8 3.1.5 Total Fluoride ........................................................................................ 9 3.1.6 Organofluorides...................................................................................... 9 3.1.7 Total Organic Carbon and Grain Size of Soil and Sediment......................... 10 3.1.8 Water Quality Parameters.......................................................................... 10 3.1.9 Bovine Fecal Samples ...............................................................................10 3.2 Biotic Sampling and Tissue Analysis........................................................................10 3.2.1 Benthic Macroinvenebrates........................................................................11 3.2.2 Mammal................................................................................................... 12 3.2.3 Fish.......................................................................................................... 13 3.2.4 Earthworm ............................................................................................... 13 3.2.5 Vegetation................................................................................................. 14 3.3 Histological assay of small mammal liver and kidney............................................ 14 . 3.4 Toxiciry Testing.................................................................................................... 14 4.0 SUMMARY OF PRELIMINARY ECOLOGICAL RISKASSESSMENT SCREEN.............. 15 5.0 DISCUSSION .................................................................................................................... 15 SECTION II 1.0 2.0 ECOLOGICAL RISK ASSESSMENT ....................................................................16 INTRODUCTION........................................................................: ...................................16 1.1 Objective............................................................................................................... 16 1.2 Site Background...................................................................................................... 16 PROBLEM FORMULATION ........................................................................................... 16 2.1 Ecological Risk Assessment ....................................................................................16 2.2 Identification of the Contaminants of Concern.........................................................17 2.3 Exposure Characterization ...................................................................................... 17 2.4 Hazard Characterization/Toxicity Assessment.......................................................... 17 2.4.1 Fluoride.................................................................................................... 17 2.4.2 Organofluorides.........................................................................................18 2.4.3 Aluminum..................................................................................................18 2.4.4 Arsenic .................................................................................................... 18 2.4.5 Beryllium.................................................................................................. 19 n 0 0 0 - ;. : USEPA 6863 000164 USFW 0581 2.4.6 Chromium.................................................................................................. 19 2.4.7 Copper...................................................................................................... 20 2.4.8 Iro n ......................................... 21 2.4.9 Lead ......................................................................................................... 21 2.4.10 Manganese ............................................................................................... 21 2.4.11 Nickel ...................................................................................................... 22 2.4.12 Vanadium..................................................................................................22 2.4.13 Zinc .........................................................................................................23 2.5 Selection of Assessment Endpoints...........................................................................23 2.6 Production of Testable Hypotheses...........................................................................24 2.7 Conceptual Model .................................................................................................. 25 2.8 Selection of Measurement Endpoints.........................................................................26 2.9 Life History/Exposure Profile Information............................................................... 29 2.9.1 The amphipod {HyalleLa azteca) as Representative of Benthic Invertebrates ................................................................................................................ 29 2.9.2 Earthworm (Eiseniafoetida) as Representative of Terrestrial Invertebrates ................................................................................................................ 30 2.9.3 Fathead Minnow (Pimephalespromelas) as Representative of FishCommunity 31 2.9.4 American Robin (Turdus migratorius) as Representative of Worm-eating Birds ................................................................................................................ 32 2.9.5 Red-tailed Hawk (Buteo jam aciertsis) as Representative of Carnivorous Birds. ...............................................................................................................33 2.9.6 Red Fox (Vulpes vulpes) as Representative of Carnivorous Mammals . . . . 34 2.9.7 Mink (Aiusiela vison) as Representative of Carnivorous M ammals..............35 2.9.8 Raccoon (Procyon lotor) as Representative of Omnivorous Mammals . . . . 37 2.9.9 Short-tailed Shrew (Biarina brevicauda) as Representative of Insectivorous M am m als................................................................................................. 38 2.9.10 Meadow Vole (Microtus penrtsylvanicus) as Representative of Herbivorous Mammals................................................................................................. 40 ASSUMPTIONS................................................................................................................. 42 EFFECTS PROFILE.......................................................................................................... 43 4.1 Fluoride.................................................................................................................. 43 m eoo4^ o o o l6 5 4.2 Organofluorides.......................................................... 4 .3 Alu m i n u m ......................................................................................... 4.4 Arsenic ..................................................................... 4.5 Beryllium'................................................................... 4.6 Chromium ................................................................. 4.7 Copper ...................................................................... 4.8 Iron .......................................................................... 4.9 Lead .......................................................................... 4.10 Manganese................................................................. 4.11 N ic k e l............................................................................... 4.12 Vanadium................................................................... 4.13 Zinc .......................................................................... 5.0 RISK CHARACTERIZATION................................................. 5.1 Benthic Invertebrate Community Structureand Function 5.2 Soil Invertebrate Community Structure and Function . . 5.3 Fish Communities ................................................... 5.4 Worm-eating Birds..................................................... 5.5 Carnivorous Birds ..................................................... 5.6 Carnivorous Mammals (Terrestrially feeding).............. 5.7 Piscivorous Mammals ................................................. 5.8 Omnivorous Mammals .............................................. 5.9 Insectivorous Mammals............................................. 5.10 Herbivorous Mammals .............................................. 6.0 UNCERTAINTY ANALYSIS ................................................. 7.0 CONCLUSIONS..................................................................... 7.1 Benthic Invertebrate Community Structureand Function 7.2 Soil Invertebrate Community Structure and Function . 7.3 Fish Communities ................................................... 7.4 Worm-eating Birds................................................... 7.5 Carnivorous Birds ....................................... .. 7.6 Carnivorous Mammals ............................................ 7.7 Piscivorous Mammals............................................... 7.8 Omnivorous Mammals ............................................ o 000166 43 44 44 44 45 45 45 46 46 46 47 47 47 48 48 48 48 48 48 49 49 49 49 50 50 50 , 51 . 51 . 51 . 51 7.9 Insectivorous Mammals...........................................................................................52 7.10 Herbivorous Mammals ...................................................................... 52 8.0 SUMMARY......................................................................................... .............................. ;2 LITERATURE C ITE D ...................................................................................................................................53 APPENDIX A Small Mammal Data S h eets................................................................................................................................... 60 APPENDIX B Analytical Reports ............................................................................................................................61 APPENDIX C Toxicity Testing Reports..................................................................................................................... 62 APPENDIX D Field N otes........................................................................................................................................ 63 APPENDIX E Statistical Analysis ............................................................................................................................ 64 000167 000 V USEPA 6866 LIST OF TABLES NUMBER 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 T E IL E Concentration of BNA's in Water Concentration of BNA's in Soil Concentration of BNA's in Sediment Concentrations of Metals in Water Results of Concentrations of Metals in Soil Results of Concentrations of Metals in Sediment Results of the Analysis for Pesticide/PCB in Water Results of the Analysis for Pesticide/PCB in Soil Results of the Analysis for Pesticide/PCB in Sediment VOA Concentrations in Water VOA Concentrations in Soil VOA Concentrations in Sediment Concentrations of Fluoride in Water Concentrations of Fluoride in Soil Concentrations of Fluoride in Sediment Results of the Organo-fluoride Analysis in Sediment Concentrations of TOC in Soil Results of the Analysis for Grain Size in Soil Concentrations of TOC in Sediment Results of the Analysis for Grain Size in Sediment In Situ Water Quality Parameters -Concentrations of Bromide, Chloride, Nitrate, Phosphorus, and Sulfate in Water VI OGO-.Ov 000168 USEPA 6867 USFW 0585 23 Concentrations of BNA's in Fecal Samples 24 Concentrations of Metals in Fecal Samples 25 Concentrations of Fluoride in Fecal Samples 25 Frequency and Abundance of Benthic Macroinvertebrates 27 Concentrations of Metals in Small Mammals 28 Concentrations of Fluoride in Small Mammals 29 Lipid Concentrations in Mammal Tissue 30 Concentrations of Metals in Fish Tissue 31 Concentrations of Puoride in Fish Tissue 32 Lipid Concentrations in Fish Tissue 33 Results of the Analysis for TAL Metals in Earthworm Tissue 34 Results of the Analysis for Fluoride in Earthworm Tissue 35 Lipid Concentrations in Earthworm Tissue 16 Concentrations of Metals in Vegetation 7 Concentrations of Fluoride in Vegetation Lipid Concentrations in Plant Tissue Results of Histopathology for the (Meadow Vole, Short-tail shrew, Meadow jumping mouse, and White-footed mouse) Summary of Toxicity Test Results Summary of Initial Risk Screen Risk Calculations Based on Wet Weight * usey^ 6S6S S1v**i'>. ` oovii 0 * 6 9 NUMBER 1 LIST OF FIGURES IITLE Sampling Site Map vm 0 0 9 -U i', USEPA 6869 000170 USFW 0587 SECTION I. TECHNICAL APPROACH, SUMMARY OF FIELD EFFORT RESULTS. AND PRELIMINARY RISK SCREEN 1.0 INTRODUCTION 1.1 Objective The objective of this project was to provide technical support to the U.S. Environmental Protection Agency Region III Removal Program in conducting an evaluation of ecological risks from alleged contamination of soil, sediment, and water at a working beef production farm located down gradient of a landfill The effort resulted in the collection of soil, sediment, surface water, and biota samples for contaminant analyses and soil, sediment, and surface water for laboratory toxicity testing. The primary goals of the project were to: 1) identify contaminants present, 2) determine the extent of contamination, and 3) produce an ecological risk assessment based on the collected data. 1.2 Site Background The site is a working beef production farm located in Washington, Wood County, WV. The owner of the farm has filed numerous complaints with the West Virginia Department of Natural Resources and the U. S. EPA alleging that contaminants are being discharged from an industrial landfill owned by the DuPont corporation, into Dry Run. Dry Run flows through the farmer's property and is a primary source of water for his cattle. The farmer maintains that numerous deaths, blindness, and other unusual illnesses observed in his herd are directly attributable to the contam inants that are discharged into Dry Run from the DuPont landfill. It has also been reported that fish and wildlife Vills have also occurred in the area, which may be associated with the abnormalities observed in the cattle. 2.0 METHODOLOGY The approach used in this document followed current U.S. EPA guidance for designing and conducting ecological risk assessments (U.S. EPA 1997). Based on the problem formulation phase of the risk assessment design, the following field study was conducted to provide data needed to complete the assessment. A screening-level ERA was conducted after the field investigation, as little data on site contamination was available prior to the effort. Numerous fish and wildlife kills, in addition to problems in the cattle, had been reported prior to this activation. 2.1 Soil Sampling Surface soil samples were collected at 4 sample areas along Dry Run and in one reference sample area (Figure 1). Sample areas were selected based on distance from the landfill outfall in an attempt to identify1a contaminant concentration gradient. Sampling was concentrated in the meadows along the stream bed. Three replicate samples were taken in each sampling area. Replicate sampling locations were determined by gridding the sampling area and randomly choosing three grid nodes for sampling through the use of a random numbers table. Sampling grid nodes were determined by usinz a random numbers table. Surface soil samples were collected using a decontaminated stainless steel trowel or spoon from the top 6 inches of the soil according to ERTC/REAC Standard Operating Procedure (SOP) *2012, Soil Sampling. All soil samples were analyzed for total organic carbon (TOC); grain size; target analyte GOO- USEPA 6870 ; 000171 USFW 058S list (TAJL) metals: TCL pesticides/PCBs; TCL Base, Neutral, and Acid Extractable (BNAs) compounds: TCL volatile organic compounds (VOCs), total fluoride, and organofluoride compounds. Additional soil was collected from the sample node closest to the stream bed for use in an earthworm toxicity test. A vegetation sample was also taken at each of the soil sampling nodes. 2.2 Sediment Sampling Sediment samples were collected at 5 sample areas on site in Dry Run, one reference sample area, and one area in Lee Creek. Sample areas were selected based on distance from the landfill outfall in an attempt to identify a contaminant concentration gradient. Sampling was concentrated in the depositional areas along the stream bed. All sediment sampling was conducted according to ERTC/REAC SOP #2016, Sediment Sampling. At each sample station, sediment was collected from the top 6 inches using a decontaminated trowel. The sample was composited into a decontaminated 5-gallon stainless steel bucket, homogenized, and divided into the appropriate sample containers for chemical analyses. Additional sediment was collected in the reference area. Tributary A, Tributary B, Area II, and Area IV, for use in a Hyalella azieca whole sediment bioassay. 2.3 Surface Water Sampling Surface water samples were collected at locations which corresponded to each of the seven sediment sample stations. Surface water samples were collected directly into two 1-liter polypropylene bottles for metals analyses and into 1-liter glass bottles for organic (i.e.,' BNAs, Pesticide/PCBs, VOCs) analyses as per ERTC/REAC SOP #2013, Surface Water Sampling. Water samples were collected prior to collecting sediment samples and upstream of any stream disturbances caused by the sampler. One sample at each location was filtered through a 0.45 micron (pm) filter in the field prior to TAL metals analysis; all the remaining TAL metals samples and all the organic samples were analyzed unfiltered. All samples analyzed for metals were preserved by adding 40 percent nitric acid until a pH of less than 2 in the sample was obtained. The filtered sample submined for TAL metal analysis was preserved after the sample was filtered. All surface water samples were submitted for TAL metals, TCL BNAs, TCL Pesticide/PCBs, TCL VOCs, chloride, fluoride, bromide, nitrate, sulfate, and phosphate analyses. Additional sample was taken in the reference area. Tributary A, Tributary B, Area II, and Area IV, for use in a Pimephaies promelas aquaeous phase toxicity test. Water quality parameters were measured using a Horiba"' water quality meter. The meter was used to measure temperature in degrees Celsius (*C), pH, dissolved oxygen [milligrams per liter (mg/L)], conductivity (millimhos per centimeter (mmhos/cm)]. oxidation reduction potential [volts (V)]. The meter was calibrated prior to and after data collection. In-situ water quality data was transcribed from the digital display of the HoribaTM into a field logbook at the time of collection. The Horiba" was used in accordance the manufacturer's operating manual. 2.4 Drinking Water Well Sampling Water was sampled from a drinking water well on the Tennant farm. Parameters were analyzed as outlined above. Samples to be analyzed were taken from a tap that was located directly on the pump head after the well had been purged for a period of approximately five minutes. 2 USEPA 6871 O O Q o.;: 000172 USFW 058! 2.5 Biological Sampling 2.5.1 Small Mammal Study Small mammals were collected from the site to determine body burden levels of TAL metals and total fluoride and to evaluate histopathological effects of exposure to site contaminants. Tissue burdens of small mammals trapped on site were compared to animal collected from the reference area. All field trapping activities were conducted in accordance with ERTC/REAC Draft Standard Operating Procedure SOP #2029, Small Mammal Sampling and Processing. Four trapping areas were established on site in areas corresponding to the soil sampling locations. A fifth grid was established on a reference area located just to the north of Dry Run in similar meadow habitat as that observed along the stream corridor (Figure 1). The reference area was chosen because the habitat present was sim ilar to that in the meadows near Dry Run, and because it was outside the area that could be directly influenced by surface water from Dry Run. The length of the trapping period and the trapping effort varied among each of the trap areas and was based on the length of time and effort required to capture a sufficient number of mammals for statistical evaluation. Sampling was performed using Museum Special snap traps set in grids. All traps were spaced 10 feet apart and baited with a rolled oats and peanut butter mixture. The traps were checked: twice daily, once in the morning and once in the evening. During trap checks, traps were rebaited and reset as necessary. Recovered animals were labeled with the trap area, trap number, species, and date of capture while in the field and then were transferred in coolers to the staging area for processing. For each animal, prior to performing the necropsy, data from the specimen label was transferred to a small mammal data sheet (Appendix A). Body metrics including total body weight, body length, tail length, ear length, liver weight, and kidney weight were measured and recorded on the data sheet. During the necropsy any abnormaliz e were noted and the contents of the gastrointestinal tract were removed from each specimen. Sections of the liver and kidney (approximately 0.5 g each) were removed for histopathological analyses. The sections were placed in a labeled 40-mL glass vial and preserved with 10 percent neutral buffered formalin. Preserved liver and kidney sections were submitted to An imal Reference Pathology (ARP) for histopathological evaluation. The remaining tissue was submitted for homogenization and TAL metal, total fluoride, percent moisture, and percent lipid analysis. 2.5.2 Vegetation Sampling Vegetation was collected by hand for residue analysis per ERTC/REAC SOP #2038 Vegetation Assessment Fie ld Protocol. The most abundant grass taxa observed at all sampling locations was targeted for residue analysis. Grass samples were taken in each area at the same grid nodes as the soil samples were taken. The above ground portion of plants from the immediate vicinity of the soil sampling node were collected by cutting the steins at the soil surface with a decontaminated knife. All samples were analyzed for TAL metals, total fluoride, percent moisture, and percent lipids. 2.5.3 Aquatic Macroinvenebrate Sampling l GO0 USEPA 6872 000173 USFW n.FQO The infaunal macroiavenebrate community was sampled per Draft ERTC/REAC SOP "2032 Bemhic Macroinvertebrate Sampling and U.S. EPA (1983, 1989, and 1990). Macroinvertebrate samples were collected for evaluation of community structure. In this investigation, macroinvertebrates were defined as organisms that impinged on a 0.5 millimeter (mm) sieve. A total of three replicates were collected from each of five sediment sampling locations (Figure 1). A long-handled, D-frame kick net, measuring approximately 45 centimeters wide and 20 centimeters tall, with 0.5 mm mesh was used. The net was used to disturb submerged vegetation and debris and collect dislodged invertebrates. Each replicate collection was performed over a uniform area at each sampling location. Benthic invertebrate samples were transferred to 500 ml polyethylene jars and preserved with a 70 percent 2-propanol solution. In the laboratory, the sample was rinsed in clean water and placed in a white 12 x 18-inch polyethylene pan withjust enough water added to allow complete dispersion of the material within the pan. Large debris, stones, and other extraneous materials were removed from the tray and inspected for attached or clinging organisms. All organisms picked from the pan were identified to the lowest positively identified taxonomic level, enumerated, and recorded on a laboratory bench sheet. The size and life history stage of the organisms and state of taxonomic knowledge of the group determined the level of identification. The organisms were identified using appropriate taxonomic references and a representative subsample were identified by a second individual to meet the Quality Assurance,'Quality Control (QA/QC) requirements of the taxonomic analysis. 2:5.4 Fish Collection Fish were collected from Dry Run to determine body burden levels of TAL metals and total fluoride. A CoffeltTMbattery powered backpack electroshocker was used and operated as per the manufacturer's instructions. The sampling team consisted of one individual operating the electroshocker and one individual collecting stunned fish with a dip net. Stunned fish were placed in a 5-gallon bucket filled with site water. Following collection, fish were identified to the lowest taxonomic level possible in the field and live specimens were released. Voucher and dead specimens were preserved with a dilute formaldehyde solution and returned to the ERT/REAC biological laboratory for confirmation of field taxonomic analyses. Fish tissue was homogenized and submined to the laboratory for TAL metal, total fluoride, % lipid and % moisture analysis. Toxicity Testing 2.6.1 Eisenia foetida (Earthworm) Toxicity Tests Five soil samples were taken for evaluation in an earthworm toxicity test. Four of the samples were taken in the meadow sampling areas along Dry Run and one in the reference meadow area as outlined above. The test was run for a period of 28 days, at which time mortality and growth in each of the test soils was enumerated. Earthworm tissue resulting from each of the treatments was submined for TAL metals, total fluoride, % lipid, and % moisture analysis. Figure 1 details the earthworm toxicity test soil sampling locations. 2.6.2 Hyalella azteca (Amphipod) Toxicity Tests USEPA 6873 4 000174 USFW 0591 Five sediment samples were taken for evaluation in an amphipod toxicity test. Four of the samples were taken in Dry Run and one in a reference area stream. The test was run for a period of 10 days, at which time mortality and growth in each of the test sediments was enumerated. Figure 1 details the amphipod toxicity test sediment sampling locations. 2.6.3 Pimepholes promelas (Minnow) Toxicity Tests Five surface water samples were taken for evaluation in a fathead minnow toxicity test. Four of the samples were taken in Dry Run and one in a reference area stream. The test was run for a period of 7 days, at which time mortality and growth in each of the test waters was enumerated. Figure 1details the fathead minnow toxicity test water sampling locations. 2.7 Sampling Equipment Decontamination The following sampling equipment decontamination procedure was employed prior to and subsequent to sampling in the following numerical sequence: 1. physical removal 2. nonphosphate detergent wash 3. potable water rinse 4. 10 percent nitric acid rinse 5. distilled water rinse 6. solvent rinse [acetone] 7. air dry 2.8 Standard Operating Procedures 2.8.1 Documentation Documentation was conducted in accordance with the following SOPs: -ERTC/REAC SOP #2002, Sample Documentation -ERTC/REAC SOP #4001, Logbook Documentation -ERTC/REAC SOP #4005, Chain o f Custody Procedures 2.8.2 Sample Packaging. Shipment, Storage, Preservation, and Handling Sample packaging, shipment, storage, preservation and handling were conducted in accordance with the following SOPs: -ERTC/REAC SOP #2003, Sample Storage, Preservation and Handling -ERTC/REAC SOP #2004, Sample Packaging and Shipment 2.8.3 Field Sampling and Analytical Techniques Field sampling activities and field analytics were conducted in accordance with the following SOPs: -ERTC/REAC SOP #2001, Cenerai Fie ld Sampling Guidelines 5 coo USEPA 6874 000175 USFW 0592 -ERTC/REAC SOP #2005, Quality Assurance/Quality Control Sample: -ERTC/REAC SOP #2006, Sampling Equipment Decontamination -ERTC/REAC SOP #2012, Soil Sampling -ERTC/REAC SOP #2013, Surface Water Sampling -ERTC/REAC SOP #2016, Sediment Sampling -REAC SOP #2029, Small Mammal Trapping and Processing -REAC SOP #2032, Benthic Sampling 2.8.4 Health and Safety Health and Safety was conducted in accordance with the following SOPs: -ERTC/REAC SOP #3001, R E A C Health and Safety Program Policy and Implementation -ERTC/REAC SOP #3012, R E A C Health and Safety Guidelines at Hazardous Waste Sites -ERTC/REAC SOP #3020, Inclement Weather, Heat Stress and Cold Stress RESULTS 3.1 Water. Soil, and Sediment Analysis 3.1.1 BNAs Surface Water Analysis of the surface water samples from Dry Run. the reference stream, and Lee Creek produced only one detection on the standard BNA scan. A sample taken in the Upper Tributary B location contained an estimated concentration of 2 ug/L of Bis(2EthylhexyOphthalate. In addition, numerous Tentatively Identified Compounds (TICs) including unknown alkane and alkene compounds were found in the surface water samples. Results for the BNA analysis of surface water samples taken in in Dry Run are presented in Table 1 and in Appendix B. Well Water The sample taken at the Tennant Farm well produced no detections from the standard BNA list. Several TICs were identified, however only one of the detected compounds could be tentatively characterized and identified as an alkene. Results for the BNA analysis of the well water sample taken at the Tennant farm is presented in Table 1 and in Appendix B. Sail Analysis of the surface soil samples from the meadows adjacent to the streambed and the reference meadow area produced a few isolated hits from the standard BNA list. Fluoranthene was detected at an estimated concentration of 23 ug/Kg in one of the three reference samples. Carbazole was detected at an estimated concentration of 41 ug/Kg in one of the three Area I soils. Di-n-burylphthalate was detected at concentrations of 22, 27, 26. and 30 ug/kg in one sample from area II, one sample from area three, and two of the three samples from area IV, respectively. Bis(2-Ethylhexyl)phthalate was detected at estimated concentrations of 27 and 62 ug/Kg in one sample from Area III and one sample 6 USEPA 6875 000 USFW 059: 'i := 000 17 6 from Area IV. Di-n-octylphthalate was detected at an estimated concentration of 180 ug/kg in one of the samples from area IV. In addition, numerous TICs including unknown alkane, cycloalkane, alkene, aldehyde, sterols, alcohols, PAH, acid, and other organic compounds were found in the surface soil samples. Results for the BNA analysis of soil samples is presented in Table 2 and in Appendix B. S.cdimcm Analysis of the sediment samples from the five site and two off-site stream locations produced only a few isolated detections of BNA compounds. Di-n-butylphthalate was detected in the Area IV sediment sample at an estimated concentration of 30 ug/Kg. Bis(2Ethylhexyl)phthalate was detected in the Area III sediment sample at an estimated concentration of 52 ug/Kg. No other standard list compounds were found in any of the Lee Creek, reference stream, or Dry Run sediment samples. Numerous TICs including unknown alkane, cycloalkane, alkene, aldehyde, sterols, alcohols, PAH, acid, and other organic compounds were found in the Dry Run, Lee Creek, and reference sediment samples. Results for the BNA analysis of sediment samples is presented in Table 3 and in Appendix B. 3.1.2 TAL Metals Surface Water Analysis of the surface water samples from Dry Run, the reference stream, and Lee Creek included both filtered and unfiltered samples for TAL metals analysis. Antimony, arsenic, beryllium, cadmium, cobalt, mercury, nickel, selenium, silver, thallium, and vanadium were not detected in any of the filtered or unfiltered water samples. Aluminum, barium, calcium, copper, iron, magnesium, manganese, potassium, sodium, and zinc were detected in the filtered water samples. Of the list of detected metals in the filtered samples, it appears that aluminum, copper, and zinc are found in higher concentrations in the Dry Run Creek drainage, including the reference stream, than in Lee Creek. Aluminum, barium, calcium, copper, iron, magnesium, manganese, potassium, sodium, and zinc were detected in the unfiltered samples. Of the list of detected metals in the unfiltered samples, concentrations of aluminum, iron, and zinc appear to be higher in Dry Run than those measured in Lee Creek. Detailed results of the TAL metals analysis in filtered and unfiltered water samples are presented in Table 4 and in Appendix B. WsiLwater Well water sampled from the well on the Tennant farm was analyzed as an unfiltered sample. Antimony, arsenic, beryllium, cadmium, chromium, cobalt, mercury, nickel, selenium, silver, thallium, and vanadium were not detected in the well sample. Concentrations measured for the remaining list of TAL metals are presented in Table 4 and in Appendix B. 7 USEPA 6876 0 0 0 ,, -J 000177 USFW0594 Soil Three replicate surface soil samples from the four Dry Run meadow areas and the reference meadow area were analyzed for TAL metals. Antimony, cadmium mercury, selenium, silver, and thallium were not detected in any of the soil samples. One-way analysis of variance determined that soil manganese concentrations were significantly higher in Area II compared to the reference area, but the same as those noted in areas I, ID. and IV (p=0.094). Area II had the highest mean manganese concentration with mean concentrations from the other areas ranging from 680 to 1310 mg/kg. Further results of the TAL metals analysis of site and reference soil samples are presented in Table 5 and Appendix B. Sediment Seven sediment samples were submined for TAL metals analysis. Five of the samples were taken in the streambed of Dry Run, one was taken in Lee Creek, and one was taken in the reference stream. Antimony, cadmium, mercury, selenium, silver, and thallium were not detected in any of the sediment samples. In comparison to the levels measured in the Lee Creek sample, it appears the Dry Run Creek reach may be enriched in aluminum, arsenic, barium, calcium, chromium, cobalt, copper, iron, lead, magnesium, manganese, nickel, potassium, sodium, vanadium, and zinc. Based on the results of the aluminum, barium, chromium, cobalt, copper, iron, lead, manganese, nickel, vanadium, and zinc analysis, there also appears to be a general trend that metal concentrations" decrease with increasing distance from the landfill. Further results of the TAL metals analysis of site and reference sediment samples are presented in Table 6 and Appendix B. 3.1.3 Pesticides/PCBs Water No pesticides or PCBs were detected in the Dry Run samples, the Lee Creek sample, or in the reference stream sample (Table 7; Appendix B). Soil No pesticides or PCBs were detected in the Dry Run meadow samples, or in the reference meadow samples (Table 8; Appendix B). Sediment No pesticides or PCBs were detected in the Dry Run samples, the Lee Creek sample, or in the reference stream sample (Table 9, Appendix B). 3.1.4 VOCs Wati No volatile organic carbon compounds were detected in the Dry Run samples, the Lee Creek sample, or in the reference stream sample (Table 10; Appendix B). 8 USEPA 6877 099.i ;v 000178 USFW 059 sail Trichlorofluoromethane was detected in every soil sample taken in the meadows adjacent to Dry Run at concentrations ranging from 0.9 to 3.6 ug/Kg. In addition, tetrachloroethene was detected in one replicate Area III soil sample at a concentration of 4.4 ug/Kg. No other volatile organic carbon compounds were detected in the Dry Run samples, the Lee Creek sample, or in the reference stream sample. Results of the VOC analysis of site and reference soil samples are presented in Table 11 and Appendix B. Sediment Acetone was detected in the Area IV sample at a concentration of 7.2 ug/Kg. Chloroform was detected at a concentration of 0.5 ug/kg in the Area III sample. No other volatile organic carbon compounds were detected in the Dry Run samples, the Lee Creek sample, or in the reference stream sample. Results of the VOC analysis of site and reference sediment samples are presented in Table 12 and Appendix B. 1.5 Total Fluoride Water / Well Water Fluoride was not detected in the Dry Run samples, the Lee Creek sample, the reference stream sample, or in the well sample taken on the Tennant farm (Table 13; Appendix B). Soil Ftuoride was detected in the Dry Run meadow and in the reference meadow samples. Soil fluoride concentrations ranged from a low of 180 mg/kg in Area IV to a high of 370 mg/kg in Area III. There appear to be no statistically significant differences in total soil fluoride concentration. Results of the soil fluoride analysis are presented in Table 14 and Appendix B. Sediment Fluoride was detected in the Dry Run creekbcd and in the reference creekbed samples, but not in Lee Creek. Fluoride concentrations ranged from a low of 290 mg/kg in the Area IV sampling area to a high of 450 mg/kg in the Upper Tributary A sampling area. Fluoride was not detected in Lee Creek. Overall, fluoride concentrations tend to decrease with increasing distance from the landfill. Sediments sampled in the Dry Run Creek reach appear to be enriched with fluoride, which is not found in Lee Creek. Results of the sediment fluoride analysis are presented in Table 15 and in Appendix B. 1.6 Oreanofluorides Sediment USEPA 6878 Because of methodology problems, specifically in obtaining appropriate standards, and the high volatility of some standards, only a limited suite of organofluoride compounds could be scanned for in the sediment samples. These compounds are presented in Table 16. Of the list that was analyzed for (Tetrafluoroethylene, hexafluoropropylene, 000179 9 000- USFW 0596 chlorodifluoromethane, perfluorocyclobutane, l-Chloro-l, 1.2.2, tetrafluoroethane, 2Chloro-l,1.1.2,3.3.-hexafluoropropane, and Perfluoroisobutylene), none of die organofluoride compounds were detected in site sediments. Results of the organofluoride analysis is reported in Table 16 and in Appendix B. 3.1.7 Total Organic Carbon and Grain Size of Soil and Sediment Summaries of total organic carbon and grain size analysis are presented in Tables 17-20 and in Appendix B. TOC in the soil ranged from an average low of 5.655 in Area III soils to an average high of 9.2 in Area IV soils. Soil grain size determinations are summarized in Table 18. TOC in the sediment ranged from a low of 1.955 in Lee Creek to a high of 4.5 55 in Area IV. Sediment grain size determination is presented in Table 20. 3.1.8 Water Quality Parameters Water quality parameters including pH. conductivity, turbidity, dissolved oxygen, temperature, bromide, chloride, nitrate, phosphate, and sulfate was measured in the Dry Run Creek reach, the reference stream, and in Lee Creek. The most notable observations were that conductivity and sulfate concentration decreased with increasing distance from the landfill. Other parameters appeared to be in the expected range. Results of these measurements and analyses are presented in Tables 21 and 22. 3.1.9 Bovine Fecal Samples ; Six fecal samples were taken to determine if environmental contaminants were showing up in the digestive products of the affected cattle. Phenol was detected in all six fecal samples at concentrations ranging from 2.6 to 8.0 mg/kg. Additionally, 4-meihylphenol was detected in all six samples in concentrations ranging from 45 to 110 mg/kg. Benzoic acid was detected in two of the samples at a concentration of 30 mg/kg. Additional information is presented in Table 23 and in Appendix B. TAL Metals Aluminum, barium, calcium, copper, iron, lead, magnesium, manganese, potassium, sodium, vanadium, and zinc were detected in the fecal samples. Additional information presented in Table 24 and in Appendix B. Fluoride Fluoride was not detected in any of the fecal samples (Table 25; Appendix B). Biotic Sampling and Tissue Analysis 3.2.1 Benthic Macroinvenebrates 10 USEPA 6879 000180 OOO.llii USFW 0597 A tool of 27 taxonomic groups were collected from the 5 locations sampled (Table 26). Of ihese, there was 1 Oligochaete, 1 Mollusc, 1Turbellarian. 3 Crustaceans, and 21 Insect taxa. Of the latter, the dominant group, in terms of taxonomic diversity, were the Coleopteria which were represented by 7 taxa. The Dipteria were represented by 4 taxa and the Ephemeropteria were represented by 3 taxa. The Plecopteria, Hemipteria Tricopteria, and Megaloptera were the least diverse groups and were represented by one or two taxa. The greatest taxonomic diversity was observed at the reference location, where 19 taxa were collected. Fewer taxa were observed at locations I through IV, and the lowest diversity was observed at location IV where 11 taxa were collected. The difference in taxonomic diversity between the reference location and locations I, II. HI, and IV was primarily due to the presence of a greater number of rare taxa at the former location. The number of individuals collected per replicate ranged from 203 at the reference location (replicate A) to 24 at location IV (replicate C). The observed density of individuals throughout the study area is primarily the result of the numerical abundance of only several taxa (Table 26). The numerically dominant taxa collected from the study area includes Leucrocuxa and Asellidae. When present, these taxa were typically the most numerically abundant organisms and were represented by 221 and 391 individuals, respectively. Other taxa. including Perlista, Chironomidae, Hyalella. the Turbellaria, and to a lesser extent, Lepiophtebia, Baeris, and Pseudolimnophilia, were present at most locations in consistently significant proportions. In general, most taxa collected were relatively rare and were represented by five or fewer individuals at most locations. For example, of the 129 total taxonomic observances, 54 were represented by one individual. 36 were represented by two to five individuals, 11 by six to 10 individuals, 15 by 11 to 20 individuals, and 13 by zreater than 21 individuals. Several taxa were collected from all locations sampled including Leucotricia, Perlesta, Chironomidae, and Asellidae (Table 26). Several taxa were not collected from all locations but were broadly represented throughout the drainage including Lepsophlebia, Agabus, Hyalella, and Turbellaria. Of the nine taxa observed at only one location, four, including Elmidae. Scinidae, Pseudolimnophila, and Stratiomvidae were collected only from the reference location. The most common distribution observed was one where a taxa was collected in relatively low numbers, and at few locations. For example. Elmidae, Hydropsyche, Limnophilidae, Nigronia, and Ceratoponidae were collected infrequently and in low numbers. Similarly, Lipogomphus, Dytiscidae, Curculidae, Elmidae, Scinidae. Histeridae, Pseudolimnophila, Stratiomyidae, and Physa were collected in low numbers at only one location. Five functional feeding groups were collected from the Dry Run drainage (Table 26). Resulting from the presence of Asellidae and Hyalella, omnivores were the dominant functional group at most locations. Although less dominant, collector-gatherers and scrapers were consistently collected from all locations in the study area and included the mayflies Leucrocuia and Leptophlebia. The dominant scraper was the mayfly Leucrocuta. Predators were dominant at locations II and III and were represented primarily by the stonefly Perlesta. Tne overall assessment of ecological condition first focuses on the evaluation of habitat quality, and secondly on the analysis of biological components in light of habitat. Habitat, 11 USEPA 6880 000- 0 000181 USFW 0598 as the principal determinant of biological potential, sets the stage for interpreting biosurvev data and can be used as a general predictor of biological condition. High quality habitat will support high quality biological communities and responses to minor alterations will be subtle and of little consequence. However, as a habitat declines in quality, discemable biological, impairment results. When habitat and biological data are systematically collected together, empirical relationships can be quantified and subsequently used for screening impact and discriminating water quality effects from habitat degradation. The watershed that drains the Dry Run study area has been modified as a result of past and present land use, particularly with respect to cattle grazing and other agricultural practices, as well as the siting of commercial and industrial facilities. The loss of riparian vegetation, through replacement by species resistant or adapted to grazing, or elimination by grazing has several consequences that should be considered when evaluating the distribution of benthic raacroinvertebrates and macroinvertebrates in the current study. The attainable biological potential of a stream or river is primarily determined by the quality of the habitat at a particular location. The Dry Run study area is situated in a rural area utilized primarily for grazing cattle and, although historic indications of grazing are evident, significant portions of the riparian area remain vegetated, and there are few areas with a completely open canopy and exposed soil. Portions of the Dry Run drainage, though somewhat degraded, support a surprisingly diverse and apparently robust aquatic community. The taxonomic diversity and numerical abundance of the macroinvertebrate was relatively high at the reference area. In contrast, the diversity and abundance at locations I. II. Ill, and IV was reduced substantially. Since habitat considerations at all locations in Dry Run are similar, the presence of contamination at the latter locations may be significant. 3.2.2 M ammal Four species of small mammals (meadow voles, short-tailed shrews, white-footed mice, and meadow jumping mice) were caught during the trapping effort. Whole bodies were submitted for lipid, TAL metal and total fluoride analysis. The trapping effort revealed at least one important field observation, which was there was extremely low trapping success in Area I, the area nearest the landfill outfall, as compared to the other areas. This is highly irregular given the similar habitats present site-wide, and may indicate an ecological threat. Field necropsies identified several significant problems with the small mammals collected in the meadow areas adjacent to Dry Run. Short-tail shrews sampled from all areas showed blackened and degenerating teeth. Shrews commonly have what is known as chestnut tipped teeth, where the extreme points of the dentitia are a light brown color. The black, mottled, and degenerating teeth observed in this study are not normally observed in shrews. One meadow vole sampled from the area was missing the left kidney, and another appeared to have and extra kidney or an extra lobe on the right kidney, independent of the adrenals. Sufficient numbers of meadow voles were caught from the Reference Area, Area II. Area III, and Area IV for statistical comparisons of tissue concentrations. Lipid concentration of meadow voles was significantly depressed in the Reference Area, Area II, and Area III, compared to that observed in Area IV (p<0.001). Barium concentration was significantly lower in the Reference Area, Area II, and Area III. compared to that observed in Area IV 12 USEPA 6881 i.T 000182 USFW 0599 (p=0.067). Sufficient numbers of short-tail shrews were caught from the Reference Area and Area III for statistical comparisons. There were no differences in body concentration of lipid. TAL metals, and fluoride in shrews taken from these two areas. Results of the trapping success, TAL metals, and fluoride analysis are presented in Tables 27-29, and in Appendix B. Results are presented by species and trapping location. 3.2.3 Fish Four species of fish were collected from Dry Run in Areas II, III, and IV. No fish were observed in Area I or the Reference Area. Creek chubs and fantail darters were collected in Area IV. Creek chubs and river chubs were collected in area III. Creek chubs, river chubs, fantail darters, central stone rollers, and black-nose dace were collected in Area II. Fish sampled during the electrofishing effort in Dry Run were submitted for whole body lipid, TAL metal and fluoride analysis. A composite sample that was taken during a historical fish kill in Dry Run was also analyzed. Since creek chubs were the only species common to all three sampling locations, statistical analysis concentrated on differentiating between tissue concentrations in this species. Aluminum, arsenic, barium, beryllium, cobalt, iron, lead, manganese, thallium, and vanadium were significantly higher in creek chubs from area II than those sample in Areas m and IV. Likewise, concentrations of these metals in Area III creek chubs were higher than those in Area IV. Conversely, cadmium and silver concentrations showed the reverse trend, with tissue concentrations in Area IV significantly higher than those measure in Areas II or III. In spite of this result, it is clear that fish inhabiting upper reaches of Dry Run. nearer to the landfill are being dosed with a significantly higher amount of metals than those in the lower reaches. There were no difference in lipid or fluoride concentration. Results of the chemical analyses are presented in Tables 30-32 and in Appendix B. 3.2.4 Earthworm Tnree replicate earthworm samples were produced from each of the earthworm toxicity test soil samples. One-way analysis of variance was used to look for significant differences in tissue concentrations between earthworms exposed to each of the five soil treatments. Cadmium and thallium concentrations in the earthworm tissue was significantly higher in the Area I exposures than those in Areas II, III, IV, or the Reference soils. Cobalt levels were lower in worms taken from the reference area, but tended to increase with increasing distance from the landfill. Copper and nickel concentrations were higher in worms taken from the reference soils than those observed in worm taken from the other soil area exposures. Further results of the earthworm tissue analyses for lipid, TAL metals and fluoride concentration are presented in Tables 33-35 and in Appendix B. 3.2.5 Vegetation 13 USEPA 6882 000- 000183 USFW 06C fluoride, aluminum, calcium, magnesium, nickel, potassium, and sodium. Further, there were strong negative associations between the growth endpoint and chromium, copper, lead, and Tine concentrations, although the relationships were not significant at the 0.10 level. Further results of the amphipod toxicity test are presented in Table 40 and in Appendix C. SUMMARY OF PRELIMINARY ECOLOGICAL RISK ASSESSMENT SCREEN Sediment, soil and water concentrations were compared against Region III BTAG screening values (U.S. EPA 1995). Hazard quotients were generated by dividing the maximum site concentration measured in each matrix by the corresponding Region III benchmark values. All contaminants for which maiimum concentrations exceeded benchmarks for sediment, soil, and water in the initial screening-level risk assessment are listed in the following sections. Contaminants that failed the initial screening process will be further evaluated in a final risk assessment for the site. Sediment Table 1 lists maximum concentrations, screening criteria, and quality criteria factors for sediment contaminants. The maximum concentration recorded at the site exceeded the benchmark values for the following compounds: arsenic, chromium, copper, manganese, and nickel. Because of the lack of a screening benchmark, the following compounds are still considered as risk factors as well: fluoride, aluminum, barium, beryllium, cobalt, iron, and vanadium. Waur Table I lists marimum concentrations, benchmarks, and quality criteria factors for water contaminants. The maximum concentration recorded at the site exceeded the benchmark values for the following compounds: aluminum, copper, and iron. Because of the lack of a screening benchmark, fluoride is considered to be a potential risk factor. Soil Table 1 lists maximum concentrations, screening criteria, and quality criteria factors for soil contaminants. The maximum concentration recorded at the site exceeded the benchmark values for the following compounds: aluminum, beryllium, chromium, copper, iron, lead, manganese, vanadium, and zinc. Because of the lack of a screening benchmark, the following compounds are still considered as risk factors as well: fluoride and trichlorofluoromethane. DISCUSSION The data generated during the field effort suggests that there are potential problems associated with conditions at the Dry Run Creek site. Minimally, the results of the sediment and water toxicity tests suggest potential problems in the stream. Some metals appear in higher tissue concentrations in biota sampled nearest the outfall of the landfill, with those levels progressively dropping with increasing distance from the landfill area. Likewise, sediments sampled in Dry Run near the landfill outfall appear to have higher fluoride and metals concentrations than those sampled further downstream. A preliminary screen of potential risk factors suggests that other problems, specifically elevated levels of fluorides, organofluorides, and some metals, may be present as well. Data gathered during the field effort will be further analyzed through a base-line ecological risk assessment for the Dry Run Creek site USEPA 6884 15 000184 000- USFW 0602 Three replicate samples of meadow grass were taken in each of the five soil sampling locations. One way analysis of variance was used to look for differences in plant tissue concentrations across the five sampling areas. Barium concentration was significantly hieher in the Area I vegetation than in the Area III and IV vegetation, but similar to that observed in plants take in the Reference and in Area II. Manganese concentration was sienificantly higher in the Area I and reference vegetation than in the Area II. Ill and IV vegetation. Cobalt was significantly higher in Area IV vegetation than that taken in any of the other areas. There was no difference in the fluoride or lipid concentration. Results of the TAL metals and fluoride analysis are presented in Tables 36-38 and in Appendix B. 3.3 Histological assay of small mammal liver and kidney Histological analysis of liver and kidney sections of meadow voles, short-tail shrews, meadow jumping mice, and white-footed mice trapped in each of the five trapping areas concluded that there were no substantive changes in liver or kidney morphology. The absence of a kidney in one animal, and the presence of an extra lobe on the right kidney of another provide anecdotal evidence of an effect, however we are unable to ascertain the importance of these observations. Full summaries of the histopathological work are presented in Table 39 and in Appendix B. 3.4 Toxicity Testing Earthworm Based on the toxicity evaluation of soils, there is no evidence for growth or survival effects on earthworms tested in any of the soil samples collected at the Dry Run Creek site. Survival was 100% in all treatment replicates and growth ranged from 32.4 to 54.3%. Further results of the earthworm toxicity test are presented in Table 40 and in Appendix C. Fathead minnow Based on the toxicity evaluation of surface water samples to the fathead minnow, it appears that surface water taken in the Upper Tributary A location induced significant mortality. Survival in the Upper Tributary A sample was 58% while survival in all other samples, including the reference location, ranged from 87 to 100%. There appear to be no growth related effects water, on the minnows in any of the water samples taken from Dry Run Creek. Survival was negatively correlated potassium concentrations, however these correlations are not statistically significant at the 0.10 level. There was a significant positive correlation between fathead survival and iron concentrations in the filtered water samples. Further results of the fathead minnow toxicity test are presented in Table 40 and in Appendix C. Amphiood Based on the results of the 10 day solid phase whole sediment toxicity test with the amphipod, Hyalella azteca, growth of the organisms was inhibited in the sediment samples taken at the Upper Tributary A, and Area II locations. There were no effects observed on survival in any of the samples taken. The observed negative growth effect was significantly negatively correlated with 14 USEPA 6883 0 0 0 - 3 000185 USFW 0601 SECTION II ECOLOGICAL RISK ASSESSMENT 1.0 INTRODUCTION 1.1 Objective The objective of this risk assessment was to provide technical support to the U.S. Environmental Protection Agency Region III in conducting an evaluation of potential ecological threat due to existing contaminant levels in soil, sediment, and water at a working beef production farm located down gradient of a landfill. Soil, sediment, surface water, and biota samples were collected for contaminant analyses and soil, sediment, and surface water were collected for laboratory toxicity testing. The information gathered during this field effort was incorporated into in an ecological risk assessment for the Dry Run Creek site. 1.2 Site Background The site is a working beef production farm located in Washington, Wood County, WV. The owner of the farm has filed numerous complaints with the West Virginia Department of Natural Resources and the U. S. EPA alleging that contaminants are being discharged from an industrial landfill owned by the Dupont corporation, into Dry Run. Dry Run flows through the farmer's property and is a primary source of water for his cattle. The fanner maintains that numerous deaths, blindness, and other unusual illnesses observed in his herd are directly attributable to the contaminants that are discharged into Dry Run from the Dupont landfill. In addition to these abnormalities, numerous fish and wildlife kills have also been reported in Dry Run since the construction of the landfill. ; 2.0 PROBLEM FORMULATION This risk assessment was designed to evaluate the potential threats to ecological receptors from exposure to site contaminants. During the preliminary risk assessment, the problem formulation process included the identification of COCs and a comparison of the maximum concentration of COCs with accepted benchmarks. This information was then used to identify complete exposure pathways of compounds exceeding benchmarks to ecological receptors and their appropriate measurement endpoints. The first step of the preliminary risk assessment process compared all chemicals analyzed in soil, sediment, and water during the field with established toxicological benchmarks. Benchmarks for sediment and soil were used to identify potential contaminants of concern for the protection of aquatic biota (U.S. EPA 1995, Long and Morgan 1990, Long et al. 1995, Persuad et al. 1992, U.S. EPA 1992, Suter and Mabrey 1994). Compounds exceeding benchmarks were retained for further evaluation using ingestion-based exposure models for higher vertebrates, and direct exposure assays for fish, benthic and terrestrial invertebrates. 2.1 Ecological Risk Assessment This ecological risk assessment was written to determine the risk associated with the exposure of biota to site-related contaminants. The following steps were completed for this preliminary risk assessment: (1) A literature search was conducted to locate life history information for selected indicator species, to determine ecotoxicological effects of site contaminants, and to locate bioconcentration factors for site contaminants. (2) An evaluation of ecological receptors was prepared. This consisted of the following: Exposure scenarios were determined based on site contaminant levels, the extent and magnitude of contamination, and the toxicological mechanisms of the 16 USEPA 6885 000186 e00.;;;5 USFW 06( contaminants. Indicator species were selected based on species present and/or potentially present on site, the availability of toxicity information from the literature, and the potential for exposure to site contaminants based on habitat use or behavior. Exposure pathway(s) were determined for each indicator species. Exposure and effect profiles were written for each indicator species and each site contaminant. A risk characterization was conducted which involved the calculation of hazard quotients (HQs) for each species for a range of exposure scenarios. Based on the results of this evaluation, the COCs identified in the initial screen were further evaluated through the use of conservative risk models. Identification of the Contaminants of Concern The contaminants ofpotential concern were identified using the initial contaminant screen. The COCs for this site that were retained through the preliminary screen include metals, fluoride, and organofluoride compounds. ; Exposure Characterization The objective of the exposure assessment is to determine the pathways and media through which receptors may be exposed to site contaminants. Potential exposure pathways are dependant on habitats and receptors present on site, extent and magnitude of contamination, and environmental fate and transport of COCs. In this base-line ecological risk assessment, it will be concluded that "a potential risk" exists ifthe HQ calculated from the maximum site concentration and the No Observed Apparent Effect Level (NOAEL) equals or exceeds l. Exposure to COCs present in forage and prey species via ingestion could cause toxicity in higher trophic level organisms. In addition to exposure via consumption of contaminated forage, ecological receptors may also be exposed through ingestion of water and incidental ingestion of soil/sediment. The exposure of benthic invertebrates, terrestrial invertebrates, and fish was determined by examining results of the toxicity tests. Hazard CharacterizatioaToxicity Assessment To determine the effects of contaminants on biota, it is necessary to understand the mechanisms of toxicity of the chemicals and the systems that they affect. Knowledge of the fate, effects, and mode of action of the COCs allows for the selection of appropriate assessment endpoints. Next is a review of the general toxicological information for the COCs identified in Section 2.4. 2.4.1 Fluoride Inorganic fluoride compounds are ubiquitous in nature. However, industrial processes such as manufacturing and mining have contributed to the environmental load of fluoride, primarily through atmospheric deposition.^Jn low doses, it is accepted that fluoride is 17 USEPA 6886 0 0 0 ..-2 G 0 0 0 1 8 7 USFW 0604 protective of teeth in humans as well as other animals. Hcm-.ver in higher levels it is generally accepted that fluoride can be toxic to both plant and animal life. Dental and skeletal lesions. lameness, stiffness of gait, appetite impairment, decreased weight gain, decreased milk production, posture abnormalities, tremors, stillbirths, overgrowth of hooves, severe diarrhea, and death have been associated with mammalian fluoride toxicity (Suttie, 1977; Shupe et al, 1992). In addition to the efFects known in mammals, birds are also susceptible to fluoride toxicity. Mortality, decreased growth rates, depressed appetite, and decreased eggshell qualityhave been reported as toxicological endpoints of fluoride exposure in birds (Fleming and Schuler 1988; Fleming et al. 1987; Guenter and Hahn 1986). 2.4.2 Organofluorides Organofluorides are used in a variety of industrial processes including the production of Teflon"*, propellants, and refrigerants. Available toxicological data generally concentrates on inhalation exposure and dermal absorption. Acute (10 day) exposure of rats to chlorodifluormethane produced decreased maternal and fetal weights, as well as an increased frequency of anopthalmia and subsequent blindness in newborn fetuses (IARC 1986). Hexafluoropropene exposure induced an increased incidence of hamster ovary cell aberrations and increased frequency grossly abnormal cells (HSDB 1997). 2.4.3 Aluminum Because of its strong reactivity, aluminum (Al) is not found as a free metal in nature. Aluminum has only one oxidation sate (+3), thus its behavior in the environment depends on its ordination chemistry and the surrounding conditions. In soils, a low pH generally results in an increase in aluminum mobility. In water, an equilibrium with a solid phase is established that controls the extent of aluminum dissolution (ATSDR 1990). Plants vary in their ability to remove aluminum from soils, although bioconcentration factors for plants are generally less than one. Biomagnification of aluminum in terrestrial food chains does not appear to occur. There is no data on the biomagnincation of aluminum in aquatic food chains (ATSDR 1990). The nervous system may be a target area for aluminum. Aluminum accumulates in neurofibrillary tangles in humans with Alzheimer's disease. Aluminum may also interact with neuronal DNA to alter gene expression and protein formation. Mammalian studies do not indicate that aluminum affects reproduction although some developmental effects have been reported in mammals (ATSDR 1990). Aluminum is known to interfere with gill transport ofoxygen and carbon dioxide in fish, and has also been identified in ionoregulatory disruption. 2.4.4 Arsenic Several review articles are available which discuss the toxic effects of As (Eisler 1988a, Nriagu 1994). Arsenic tends to be widespread in the environment (Woolson 1975) and is constantly being oxidized, reduced, or mobilized (Eisler 1938a). Physical processes are important in determining As bioavailability in aquatic environments. For example, arsenates are readily adsorbed onto sediments with high organic matter, and arsenates are more strongly adsorbed onto sediments than other As forms. However, absorption depends on the As concentration, sediment characteristics, pH, and ionic concentration of other compounds USEPA 6887 <i t O l *-J 000188 USFW060 (Eisler 1988a; U.2. U4. 19811. The U.S. EPA (1981) noted that arsenate (pentavalent) is the predominant As form in oxygeu_;= rnd that arsenite (trivalent) is the predominant As form in anaerobic conditions. Arsenic is not significantly concentrated in aquatic invertebrates; whole body concentration factors for invertebrates range from 3 to 17 for exposure to arsenic trioxide (trivalent) and from 0 to 7 for arsenic pentoxide (pentavalent). Arsenic may be bioconcentrated by organisms at the bottom of the food chain; however, data do not indicate that significant biomagnification occurs (U.S. EPA 1985). 2.4.5 Beryllium The majority of the beryllium (Be) in the environment is the result of coal and oil combustion. Beryllium naturally enters waterways through the weathering of rock and soil, and through deposition of atmospheric beryllium. Upon reaching water and soil, beryllium is most likely retained as an insoluble form that is generally immobile. However, beryllium chloride, fluoride, nitrate, phosphate, and sulfate (tetrahydrate) are all water-soluble forms (ATSDR 1993a). Due to its geochemical similarity to aluminum, beryllium may be expected to adsorb onto clay surfaces at low pHs, and it may remain precipitated as insoluble complexes at higher pHs. Therefore, beryllium is expected to have limited mobility in soil (ATSDR 1993a). ; Beryllium is not expected to bioconcentrate in aquatic animals and no evidence for significant biomagnificarion within food chains has been found. Beryllium is extremely toxic to warmwater fish in soft water. The degree of toxicity decreases with increasing hardness (ATSDR 1993a). Major exposure routes for aquatic ecological receptors include ingestion of contaminated soil and sediment. Although several studies point out the negative effects of beryllium in mammalian systems, no studies that evaluated the relationship between sediment beryllium concentration and observed toxicity to benthic organisms could be found (ATSDR 1993a). 2.4.6 Chromium Chromium (Cr) can exist in oxidation states ranging from -2 to +6, but is most frequently converted to the relatively stable trivalent (+3) and hexavalent (--6) oxidation states (Eisler 1986a). In both freshwater and marine systems, hydrolysis and precipitation are the most important processes that determine the fate and effects of Cr, whereas adsorption and bioaccumulation are relatively minor. Precipitated Cr*1hydroxides remain in sediments under aerobic conditions. However, under anoxic and low pH conditions, Cr*1hydroxides may solubilize and remain as ionic Cr*' unless oxidized to Cr*4through mixing and aeration (Eisler 1986a). In soils, the solubility and bioavailability of Cr are governed by soil pH and organic complexing substances, although organic complexes play a more significant role (James and Bartlett 1983a; James and Bartlett 1983b). Th* trivalent state is the form usually found in biological materials. This form functions as an essential element in mammals by maintaining efficient glucose, lipid, and protein metabolism (Stevens et al. 1976). Chromium is beneficial but not essential to higher plants (Eisler 1986a). The biomagnification and toxicity of Cr*1is low relative to Cr*4because of 19 USEPA 6888 000189 /> -r. ' 1 O USFW 0606 its low membrane permeability and its noncorrosivity. However, a large degree of accumulation by aquatic and terrestrial plants and animals in the lower trophic levels has been documented (Eisler 1986a), although, the mechanism of accumulation remains largely unknown. Chromium is mutagenic, carcinogenic, and teratogenic, with Cr"4 exhibiting the greatest toxicity; relatively less is known about the toxicity of Cr*1. At high concentrations" Cr*4 is associated with abnormal enzyme activity, altered blood chemistry, lowered resistance to pathogenic organisms, behavioral modifications, disrupted feeding, histopathology, osmoregulatory upset, alterations in population structure, and inhibition of photosynthesis. Rabbits fed dietary Cr accumulated hyaluronates, chondroitin sulfates, and neutral mucopolysaccharides in the soft tissues, causing pericapillary sclerosis (Kucher and Shabanov 1967). This accumulation blocked blood tissue barriers, which are permeable under normal conditions, preventing the normal transport of metabolites. One manifestation of this condition was the inhibition of insulin production in the pancreatic islets due to damage to the beta-cells contained therein. Chromium also leads to nephron damage via swelling and loss of microvilli, the formation of intracellular vacuoles, mitochondrial swelling, and cytoplasmic liquefication and loss of cells lining the nephron surface (Evan and Dail 1974). The preliminary step in Cr-induced respiratory cancer is speculated to be the scarring of alveolar tissue, followed by the elicitation of inflammatory reactions in lung tissue leading to bronchopneumonia, alveolar epithelial changes, atrophy, and benign tumor formation. Direct skin contact with highly corrosive chromic acid and its anhydride produces skin ulcers and necrosis by a mechanism independent of any allergic response (Steven et al. 1976). 2.4.7 Copper Copper (Cu) does not appear to have mutagenic properties (IRIS 1990), but it is a teratogen (RTECS 1991) and a possible carcinogen (Venugopal and Luckey 1978). Copper is caustic, and acute toxicity is primarily related to this property (Hatch 1978). Copper is an essential element for animals and is a component of many metalloenzymes and respiratory pigments (Demayo et al. 1982). It is also essential to iron (Fe) utilization and functions in enzymes for energy production, connective tissue formation, and pigmentation (Venugopal and Luckey 1978). Excess Cu ingestion leads to accumulation in tissues, especially in the liver. High levels of Cu modify hepatic metabolism (Brooks 1988), which may lead to inability of the liver to store and excrete additional Cu. When liver concentration exceeds a certain level, the metal is released into the blood, causing hemolysis and jaundice. High Cu levels also inhibit essential metabolic enzymes (Demayo et al. 1982). Toxic symptoms appear when the liver accumulates 3 to 15 times the normal level of Cu (Demayo et al. 1982). Although the exact mechanism oftoxicity is not known, the following mechanisms have been proposed: formation of stable inhibitory complexes with cytochrome P-450 (Wiebel et al. 1971); impairment of function ofNADPH-cytochrome c reductase and alteration of mixed function oxidations (Reiners etal. 1986); and inhibition of heme biosynthesis (Martell 1981). Intranuclear inclusions may act as a detoxifying mechanism where Cu is compiexed by 20 USEPA 6889 000190 0 0 0 -i '0 USFW 060-, protein ligands, protecting cytoplasmic organelles (Demayo et al. 1982). Ruminants are the most sensitive mammal species to Cu toxicosis. Young animals retain more dietary Cu than older animals and are more sensitive to Cu toxicity (Venugopal and Luckey 1978). 2.4.8 Iron Iron (Fe) is commonly detected in concentrations of 5 percent or more in soil. It is used primarily in the production of steel and other alloys as well as a major source of hydrogen. Iron is a constituent of hemoglobin and is essential to plant and animal life as well as being an important component in cellular oxidative processes. The disposition of ingested iron is regulated by a complex mechanism to maintain homeostasis. Therefore, bioconcentration in biota is not expected to be a significant process for iron. Generally, about 2 to 15 percent of ingested iron is absorbed from the gastrointestinal tract, and elimination is approximately 0.01 percent of the body burden per day. Adverse effects of iron toxicity may include renal failure and hepatic cirrhosis. The mechanism of toxicity begins with acute mucosal cell damage and absorption of ferrous ions directly into circulation, resulting in capillary endothelial cell damage to the liver (Shacklette and Boemgen 1984). 2.4.9 Lead Lead does not biomagnify to a great extent in food chains, although accumulation by plants and animals has been extensively documented (Wixson and Davis 1993, Eisler 1988b). Older organisms typically contain the highest tissue Pb concentrations, with the majority of the accumulation in the bony tissue of vertebrates (Eisler 1988b). Predicting the accumulation and toxicity of Pb is difficult since its effects are influenced to a very large degree, relative to other metals, by interactions among physical, chemical, and biological variables. In general, organolead compounds are more toxic than inorganic Pb compounds, and young, immature organisms are most susceptible to its effects (Eisler 1988b). In plants, Pb inhibits growth by reducing photosynthetic activity, mitosis, and water absorption. The mechanism by which photosynthetic activity is reduced is attributed to the blocking of sulfhydryl groups, inhibiting the conversion of coproporphyrinogen to prcporphyrinosen (Holl and Hampp 1975). The toxic effects of Pb on aquatic and terrestrial organisms are extremely varied and include mortality, reduced growth and reproductive output, blood chemistry alterations, lesions, and behavioral changes. However, many effects exhibit general trends in their toxic mechanism. Generally, Pb inhibits the formation of heme, adversely affects blood chemistry, and accumulates at hematopoietic organs (Eisler 1988b). At high concentrations near levels causing mortality, marked changes to the central nervous system occur prior to death (Eisler 1988b). Plants can uptake Pb through surface deposition in rain, dust, and soil, or by uptake through the roots. The ability of a plant to uptake Pb from soils is inversely related to soil pH and organic matter content. Lead can inhibit photosynthesis, plant growth, water absorption. 2.4.10 Manganese USEPA 6890 000191 USFW 06( soluble pentavalent form. The mobility of vanadium in soil is affected by pH, redox potential, and the presence of particulates. Relative to other minerals, vanadium is mobile in neutral or alkaline soils and its mobility decreases in acidic soils (ATSDR 1991; Van Zinderen Bakker and Jaworski 1980). In the terrestrial systems, bioconcentration is more common in lower plant species. In addition, vanadium concentrations in plants are dependent on the amount of water-soluble vanadium, pH, and growing conditions. Vanadium appears to be present in all terrestrial mammals but the concentrations are usually below the detection limits. The highest concentration of vanadium is usually found in the liver and skeletal tissues (ATSDR 1991). Vanadium is very poorly absorbed into the gastrointestinal tract and the toxic mechanism of vanadium on the respiratory system is similar to other metals (Castronova et al. 1984). Vanadium damages the alveolar macrophages by decreasing the macrophage membrane integrity. Damaged macrophages inhibit the ability of the respiratory system to clear itself of other particles. In vitro experiments indicate that the mechanism of toxicity of vandium is by inhibiting sodium-potassium ATPase activity, which inhibits the sodium-potassium pump. This pump is necessary for the transport of material across cell membranes (Nechay and Saunders 1978). 2.4.13 Zinc Zinc (Zn) is essential for normal growth and reproduction in plants and animals and is regulated by metallothioneins. Metallothioneins act as temporary Zn storage sites and aid in reducing the toxicity ofZn to both vertebrates and invertebrates (Olsson et al. 1989). Zinc is not known to bioaccumulate in food chains, because it is regulated by the body and excess Zn is eliminated. Zinc has its primary metabolic effect on Zn-dependant enzymes that regulate the biosynthesis and catabolic rate ofribonucleic (RNA) acid and deoxyribonucleic acid (DNA). High levels of Zn induce Cu deficiency and interfere with metabolism of calcium (Ca) and Fe (Goyer 1986). The pancreas and bone seem to be the primary targets of Zn toxicity in birds and mammals. Pancreatic effects include cytoplasmic vacuolation, cellular atrophy, and cell death (Lu and Combs 1988, Kazacos and Van Vleet 1989). Zinc preferentially accumulates in bone, and induces osteomalacia (a softening of bone caused by a deficiency of Ca, phosphorus and other minerals) (Kaji et al. 1988). Gill epithelium is the primary target site in fish. Zinc toxicosis results in destruction of gill epithelium and tissue hypoxia (Spear 1981). Selection of Assessment Endpoints The information gathered during a site reconnaissance and during the field work, and subsequent discussions with the U.S. EPA on-scene coordinator and the Region III Biological Technical Assistance Group, allowed for the selection of assessment endpoints that corresponded to the habitat types present at the Dry Run Creek site. The site is composed of a variety of habitats including forested and old-field uplands, grassy meadows, the creek, and associated riparian areas. A variety of birds, mammals, and fish may use the site for feeding and nesting. Likewise, terrestrial and benthic invertebrates are key elements in the functions of these systems. Therefore, the assessment endpoints focused toward these faunal groups. Viability of terrestrial, avian, and aquatic populations and organism survivability were selected as assessment endpoints for this risk assessment. Listed next are 000-':,? 2 000192 USEPA 6892 USFW 0610 Manganese (Mn) does not occur as a free metal in the environment but is a component of numerous minerals. Elemental manganese and inorganic manganese compounds have negligible vapor pressures, but may exist in air as suspended particulate matter derived from industrial emissions or the erosion of soil. Removal from the atmosphere is mostly through aravitational settling. The transport and partitioning of manganese in water is controlled by the solubility of the specific chemical form present. The metal may exist in water in any of four oxidation states (2+, 3+, 4+, or 7+). Divalent manganese (Mn-*-2) predominates in most waters (pH 4 to 7), but may become oxidized at a pH greater than 8 or 9. Manganese is often transported in moving water as suspended sediments. The tendency of soluble manganese compounds to adsorb to soils and sediments depends mainly on the cation exchange capacity and the organic composition of the soil. Manganese in water may be significantly bioconcentrated at lower trophic levels. However, biomagnification in the food chain may not be significant (ATSDR 1990). The amount of manganese absorbed across the gastrointestinal tract is variable. There does not appear to be a marked difference between manganese ingested in food or in water. One of the key determinants of absorption appears to be dietary iron intake, with low iron levels leading to increased manganese absorption. This is probably because both iron and manganese are absorbed by the same transport system in the gut (ATSDR 1990). 2.4.11 Nickel Pure nickel (Ni) is a hard, white metal that is usually used in the formation of alloys (such as stainless steel) and Ni combined with other elements is found in all soils. Nickel is the 24'*' most abundant element and is found in the environment as oxides or sulfides. Nickel may be released into the environment through mining, oil-burning power plants, coal-burning power plants, and incinerators. Nickel will attach to soil or sediment panicles, especially those containing Fe or manganese (Mn). Under acidic conditions, Ni may become more mobile and seep into the groundwater. The typical Ni concentration reported in soils is from 4-80 milligrams per kilograms (mg/kg). The speciation and physicochemical state of Ni is important in considering its behavior in the environment and its availability to biota. The most probable exposure routes of Ni are through dermal contact, inhalation of dust, and ingestion of Ni-contaminated soil. The respiratory system is the primary target of Ni exposure following inhalation. Manifestations such as inflammation of the lungs, fibrosis, macrophage hyperplasia, and increased lung weight have been noted in animals exposed to Ni. Animals exposed to Ni through oral exposure were noted to have lethargy, ataxia, irregular breathing, salivation, and squinting (ATSDR 1996). 2.4.12 Vanadium Elemental vanadium does not occur naturally but it can exist in 50 different ores and fossil fuels. Other anthropogenic sources include acid-mine leachate, sewage sludge, and fertilizers. The principal use of vanadium is as an alloy constituent, especially in steel. The addition of vanadium to steel removes oxygen and nitrogen, which improves the strength. The average concentration of vanadium in the earths crust is 150 mg/kg and in the U.S. soils are 200 mg/kg (Byemum et al. 1974). The release of vanadium to water and soil occurs as a result of the weathering of rocks and from soil erosion. This process usually converts the less-soluble trivalent form to the more- 22 USEPA 6891 000193 000- / i USFW 060! on the site? Are levels of site contaminants sufficient to cause negative impacts on growth, survival, and reproductive success of carnivorous birds due to the ingestion of contaminated forage, soil, and water on the site? Are levels of site contaminants sufficient to cause negative impacts on growth, survival, and reproductive success of carnivorous mammals due to the ingestion of contaminated forage, soil, and water on the site? Are levels of site contaminants sufficient to cause negative impacts on growth, survival, and reproductive success of piscivorous mammals due to the ingestion of contaminated forage, soil, and water on the site? Are levels of. site contaminants sufficient to cause negative impacts on growth, survival, and reproductive success of omnivorous mammals due to the ingestion of contaminated forage, soil, and water on the site? Are levels of site contaminants sufficient to cause negative impacts on growth, survival, and reproductive success of insectivorous mammals due to the ingestion of contaminated forage, soil, and water on the site? Are levels of site contaminants sufficient to cause negative impacts on growth, survival, and reproductive success of herbivorous mammals due to the ingestion of contaminated forage, soil, and water on the site? Conceptual Model The conceptual model relies on contaminant and habitat characteristics to identify critical exposure pathways to the selected measurement endpoints. At the Dry Run Creek site, contaminants in the soil may come in contact with subsurface (earthworms) and above-ground terrestrial receptors (small mammals) inhabiting the wooded, wetland, and open field areas of the site. Subsurface terrestrial receptors in these areas may be exposed to site contaminants through direct contact with the soil, and in some cases, the intentional ingestion of soil. Above-ground terrestrial receptors may be exposed to contaminants through direct contact with the soil, the ingestion of subsurface terrestrial organisms, the ingestion of other above-ground terrestrial receptors, the incidental ingestion of soil adhered to food items, and the intentional ingestion of surface water from any of the on-site surface drainages. The wooded areas, riparian area, and meadow areas provide distinct habitat types that may support a variety of terrestrial and avian receptors. For example, a small omnivorous mammal may occupy one or all the habitat types, whereas, an individual carnivorous mammal may regularly traverse all three habitats daily in search of food items. Avian piscivores and carnivores may be exposed to site contaminants in much the same way as an above-ground terrestrial receptor. The consumption of contaminated prey, the incidental ingestion ofsoil/sediment, and the consumption ofsurface water may transfer contaminants to these receptors. The conceptual model relies on contaminant and habitat characteristics to identify critical exposure pathways to the selected measurement endpoints. The preliminary risk screen identified metals, fluoride, and trichlorofiuoromethane as the primary contaminants exceeding benchmarks in site sediment, soil, and water. Benthic macroinvertebrates, fish, and terrestrial invertebrates may be 25 USEPA 6894 0 0 0 34 000194 USFW 0612 the specific assessment endpoints selected for this ecological risk assessment. Ten assessment endpoints were chosen to evaluate the risk of contaminants at the Dry Run Creek site: 1) protection of benthic invertebrate community structure and function 2) protection of soil invertebrate community structure and function 3) protection of fish communities to ensure that direct exposure to contaminants does not have a potential negative impact on growth, survival, or reproductive success. 4) protection of worm-eating birds to ensure that ingestion of contaminants in forage does not have a potential negative impact on growth, survival, or reproductive success. 5) protection of carnivorous birds to ensure that ingestion of contaminants in forage does not have a potential negarive impact on growth, survival, or reproductive success. 6) protecrion ofcarnivorous mammals to ensure that ingestion of contaminants in forage does not have a potential negative impact on growth, survival, or reproductive success. 7) protection ofpiscivorous mammals to ensure that ingestion of contaminants in forage does not have a potential negative impact on growth, survival, and reproductive success. 8) protection ofomnivorous mammals to ensure that ingestion of contaminants in forage does not have a potential negative impact on growth, survival, and reproductive success. 9) protection of insectivorous mammals to ensure that ingestion of contaminants in forage does not have a negative impact on growth, survival, and reproductive success. 10) protection of herbivorous mammals to ensure that ingestion of contaminants in forage does not have a negative impact on growth, survival, and reproductive success. Production of Testable Hypotheses The testable hypotheses are specific risk questions that are based on the assessment endpoints. Based on the mechanism of contaminant toxicity, the number of exposure pathways that may exist for an assessment endpoint, or other factors, there may be more than one question for each assessment endpoint. Are levels of site contaminants sufficient to have negative effects on benthic invertebrate community structure and function? Are levels of site contaminants sufficient to have negative effects on soil invertebrate community structure and function? Are levels of site contaminants sufficient to cause direct toxicity to fish growth, survival, and reproductive success? Are levels of site contaminants sufficient to cause negative impacts on growth, survival, or reproductive success of worm-eating birds due to the ingestion of contaminated forage, soil, and water 24 USEPA 6893 0 0 0 <-3 <7J 000195 USFW 061 a) Ingestion of vegetation b) Incidental ingestion of sediment c) Incidental ingestion of water 8 Selection of Measurement Endpoints Measurement endpoints are measurable ecological characteristics that are related to the valued characteristics selected as assessment endpoints. Measurement endpoints should be linked to the assessment endpoints by the mechanism oftoxicity and the route of exposure. Measurement endpoints are used to derive a quantitative estimate of potential effects, and form a basis for extrapolation to the assessment endpoints. Measurement endpoints were selected on the basis of potential presence of receptors on site, and the potential for exposure to contaminants of concern. The availability of the appropriate toxicity information on which risk calculations could be based was also an important consideration. Endpoints selected were determined to be representative of exposure pathways and assessment endpoints identified for the site. Next is a list of specific measurement endpoints that correspond to the assessment endpoints identified in Section 2.5. Measurement endpoints for assessment endpoint: protection of benthic invertebrate communities structure and function To evaluate the structure and function of the benthic community, benthic macroinvertebrates were collected from five locations in Dry Run. Existing community structure was evaluated at each of the five locations by determining taxonomic diversity and through an evaluation of functional feeding groups. Sediment was also collected in each of the five areas for toxicity testing using the amphipod, Hyallela azteca. Tne endpoints of these tests were survival and growth. Collocated sediment samples were also collected and analyzed for target analyte list (TAL) metals. BNA's, Pest/PCBs, VOCs, fluoride, grain size, and total organic carbon (TOC). The chemistry results were then correlated with observed adverse biotic responses in the toxicity tests in order to determine risk potential. Measurement endpoints for assessment endpoint: protection of soil invertebrate community structure and function To evaluate the structure and function of the benthic community, soil was collected from each of the meadow locations and tested using the earthworm, Eiseniafoetida in toxicity tests. The endpoints of these tests were survival and growth. Collocated soil samples were also collected and analyzed for target analyte list (TAL) metals, BNA's, Pest/PCBs, VOCs, fluoride, grain size, and total organic carbon (TOC). Measurement endpoints for assessment endpoint: protection of fish communities to ensure that direct exposure to contaminants does not have a negative impact on growth, survival, and reproductive success. 27 U SEPA 6896 0G0A3S 000196 I |Q cr\A / n e - i a exposed to contaminated sediment, water, or soil through direct toxicity. For the purposes of this risk assessment, the concentration ofthe contaminants ofconcern found in the sediment, water, or soil were correlated with toxicity levels identified in the corresponding toxicity tests to determine if benthic invertebrates fish, or terrestrial invertebrates may be at risk. Terrestrial receptors may be exposed to contaminants by feeding on organisms which have accumulated COCs in their tissues. Higher trophic level receptors may also be exposed to contaminants from food ingestion and via incidental ingestion of soil/sediment and water. The pathway to the reference area meadow is unknown, however the pathway to the reference area stream may involve groundwater transport. The following pathways were evaluated in this risk assessment: I. Benthic invertebrates a) Direct exposure to sediment II. Soil invertebrates a) Direct exposure to soil III. Fish a) Direct exposure to water IV. Worm'eating bird a) Ingestion of earthworms b) Incidental ingestion of soil 0 Incidental ingestion of water V. Carnivorous bird a) Ingestion of small mammals b) Incidental ingestion of soil c) Incidental ingestion of water VI. Carnivorous mammal a) Ingestion of small mammals b) Incidental ingestion of soil c) Incidental ingestion of water VII. Piscivorous mammal a) Ingestion of forage fish b) Incidental ingestion of sediment c) Incidental ingestion of water VIII. Omnivorous mammal a) Ingestion of forage fish b) Incidental ingestion of sediment c) Incidental ingestion of water IX. Insectivorous mammal a) Ingestion of earthworms b) Incidental ingestion of sediment c) Incidental ingestion of water X. Herbivorous mammal 26 0 r, n t USEPA 6895 000197 USFW 0613  Protection of omnivorous mammals to ensure that ingestion of contaminants in forage does not have a negative impact on growth, survival, and reproductive success. Food chain accumulation studies were selected to evaluate risk to mammalian species which utilize the site and adjacent areas. The selected measurement endpoint receptor species is the raccoon. Procyan lotor, as a model for omnivorous mammalian species. Appropriate forage species (fish) were identified for the above receptors and the dietary exposure of receptors to contaminants was quantified. Measurement endpoints for assessment endpoint: Protection of insectivorous mammals to ensure that ingestion of contaminants in forage does not have a negative impact on growth, survival, and reproductive success Food chain accumulation studies were selected to evaluate risk to mammalian species which utilize the site and adjacent areas. The selected measurement endpoint receptor species is the short-tail shrew, Blarina brevicauda, as a model for insectivorous mammalian species. Appropriate forage species (earthworms) were identified for the above receptors and the dietary exposure of receptors to contaminants was quantified. Measurement endpoints for assessment endpoint: Protection of herbivorous mammals to ensure that ingestion of contaminants in forage does not have a negative impact on growth, survival, and reproductive success Food chain accumulation studies were selected to evaluate risk to mammalian species which utilize the site and adjacent areas. The selected measurement endpoint receptor species is the meadow vole, Microtuspenrtsylvanicus, as a model for herbivorous mammalian species. Appropriate forase species (vegetation) was identified for the above receptors and the dietary exposure of receptors to contaminants was quantified. Life History/Exposure Profile Information Receptor species were selected from several trophic levels. Organisms which were likely to be exposed to contaminants because of specific behaviors, patterns of habitat use. or feeding habits were selected for evaluation in this risk assessment. The availability of appropriate toxicity information on which risk calculations could be based was also an important consideration. The terrestrial invertebrate receptor selected for this assessment is the earthworm. The terrestrial vertebrate receptor species selected for this risk assessment are: meadow vole, short-tail shrew, raccoon, mink, and red fox. The avian receptor species selected for this risk assessment are: American robin and red-tailed hawk. The aquatic vertebrate receptor species for this risk assessment is the fathead minnow. The aquatic invertebrate receptor is H. azteca. 2.9.1 The amphipod (Hyallela azteca) as Representative of Benthic Invertebrates Justification Hyallela azteca was selected as representative of benthic invertebrates due to their direct contact with sediment for a significant portion of their life cycle, ubiquitous distribution in aquatic systems, importance as a food item for aquatic-invertebrate consumers, and ease of 29 USEPA 6898 000 '^000198 Fathead minnow, Pimephalespromelas. toxicity tests were used to determine the toxicity of the water in Dry Run. The endpoints of these tests were survival and growth. Collocated water samples were also collected and analyzed for target analyte list (TAL) metals, BNA's, Pest/PCBs, VOCs, fluoride, grain size, and total organic carbon (TOC). The chemistry results were then correlated with observed advene biotic responses in the toxicity tests in order to determine risk potential. Measurement endpoints for assessment endpoint: Protection of worm*eating birds to ensure that ingestion of contaminants in forage does not have a negative impact on growth, survival, and reproductive success. Food chain accumulation studies were selected to evaluate risk to avian species which utilize the site as a feeding area. The selected measurement endpoint receptor species is the American robin, Turdus migratorius. Appropriate forage species (earthworms) were identified for the above receptor, and the dietary exposure of receptors to contaminants was quantified and compared to existing toxicity data for these, or other closely related species. Measurement endpoints for assessment endpoint: Protection of carnivorous birds to ensure that ingestion of contaminants in forage does not have a negative impact on growth, survival, and reproductive success. Food chain accumulation studies were selected to evaluate risk to avian species which utilize the site as a feeding area. The selected measurement endpoint receptor species is the red-tailed hawk, Buteo jamaciensis. Appropriate forage species (small mammals) were identified for the above receptor, and the dietary exposure of receptors to contaminants was quantified and compared to existing toxicity data for these, or other closely related species. Measurement endpoints for assessment endpoint: Protection of carnivorous mammals to ensure that ingestion of contaminants in forage does not have a negative impact on growth, survival, and reproductive success. Food chain accumulation studies were selected to evaluate risk to mammalian species which utilize the site and adjacent areas. The selected measurement endpoint receptor species is the red fox, Vulpes vulpes. Appropriate forage species (small mammals) were identified for the above receptors and the dietary exposure of receptors to contaminants was quantified. Measurement endpoints for assessment endpoint: Protection of piscivorous mammals to ensure that ingestion of contaminants in forage does not have a negative impact on growth, survival, and reproductive success. Food chain accumulation studies were selected to evaluate risk to mammalian species which utilize the site and adjacent areas. The selected measurement endpoint receptor species are the mink. Mustela vison. Appropriate forage species (fish) were identified for the above receptors and the dietary exposure of receptors to contaminants was quantified. Measurement endpoints for assessment endpoint: USEPA 6897 000199 000--..: V USFW061 earthworms were observed in both the wooded and open field areas of the Dry Run Creek site. Lift Histcn Earthworms feed on dead and decaying plant and animal remains and on free-living soil microflora and microfauna. Their primary source of food is dead plant material, especially plant liner. Next to food, their most important requirement is adequate moisture. Water conservation mechanisms are poorly developed; respiration depends on diffusion of gases through the body wall which must be kept moist. Earthworms are generally absent or rare in soiis with very coarse texture, in soils with high clay content in regions of high rainfall, and in soils with a pH of less than 4 (Lee 1985). Earthworms are hermaphroditic and most species reproduce by cross-fertilization, although many species can also produce cocoons parthenogenetically. Sexual reproduction cannot occur without a ditellum, ovaries, oviducts, and possibly the ovisacs, but male organs are not essential. The population of an earthworm species at any one time consists of young immature, well-grown immature (adolescent), mature, and senescent individuals (Edwards and Lofty 1977). Earthworms have several ways of surviving adverse environmental conditions such as soil desiccation and ambient cold and heat In terms of population survival, the cocoons can resist desiccation and temperature extremes much more easily than mature individuals. Worms may also migrate to deeper soil or undergo states of inactivity until environmental conditions become favorable once again (Edwards and Lofty 1977). Some species of worms grow throughout their lives by continually adding segments proliferated from a growing zone locatedjust in front of the anus. Other species, such as . foetida, possess the adult number of segments upon hatching and increase in size without increasing the number of segments. The life span of Eisenta foetida was reported to be approximately 4.5 years under laboratory conditions (Edwards and Lofty 1977). Exposure Profile Direct contact with contaminated soil is the primary route of exposure for earthworms in this risk assessment. Survival and growth endpoints following exposure to site soils will be used to evaluate risk to these organisms. Tissue residue analysis will also be conducted on the worms to determine exposure to higher trophic level organisms. .9.3 Fathead Minnow (Pimephales promelas) as Representative of Fish Community Justification The fathead minnow was selected as representative of omnivorous fish due to its dietary composition, direct contact with water throughout the life cycle, ubiquitous distribution in aquatic systems, importance as a food item for fish-eating consumers, and ease of use in laboratory toxicity evaluations. L ift Histotv USEPA 6900 31 0 0 0 - 0 000200 I to m * / use in laboratory toxicity evaluations. These species are also likely to occur in the surface sediment at the Dry Run Creek site. 1ife History (Hvalleta azteca) The amphipod, Hyallela azteca, is commonly found in freshwater lakes, streams, ponds, and rivers throughout North and South America. In preferred habitats, they are known to reach densities in excess of 10,000 per square meter. They may also be found in sloughs, marshes, and ditches, but generally in lower numbers (U.S. EPA 1994). Hyallela azteca are epibenthic detritivores that feed on coarse paniculate organic material. They typically burrow into surface sediment, and avoid bright light. Because oftheir feeding and behavioral characteristics, they are ideal test organisms for toxicological evaluation of freshwater sediments. Avoidance of light by movement into the sediment keeps these organisms almost constantly in contact with sediment contaminants (U.S. EPA 1994). Reproduction in this crustacean is sexual. Males are larger than females and have larger front snathopods that are presumably used for holding the female during amplexus and copulation. During amplexus, the male and female feed together for a period of up to one week. The pair separates temporarily while the female goes through a molting period. Immediately after the molt, the two rejoin and copulation begins. During copulation, the male releases sperm near the female's marsupium. The female sweeps the sperm into her marsupium. and simultaneously releases eggs from her oviducts, into the marsupium, where fertilization take:s place. The "average brood size for female Hyallela azteca is 18 eggs per brood, but this number can vary with environmental conditions and physiological stress (U.S. EPA 1994). Developing embryos and hatched young are kept inside the female's marsupium until she undergoes a second molt. At that time, the juvenile Hyallela azteca are released into the surrounding environment. Under favorable conditions, each female produces approximately one brood during every ten day time period (U.S. EPA 1994). Hyallela azteca have a minimum of 9 instars, with 5 to 8 pre-reproductive stages. Tne first five stages arejuvenile stages; instars 6 and 7 form the adolescent stages; and stages 8 and higher are considered adult (fully reproductive) stages (U.S. EPA 1994). Exposure Profile for Hvallela azteca Since direct contact with contaminated sediment in the toxicity evaluation is the primary route of exposure for Hyallela azteca in this risk assessment, the results of the test will be used to indicate exposure. .9.2 Earthworm (Eiseniafoetida) as Representative of Terrestrial Invertebrates Justification Earthworms were selected as representative of terrestrial invertebrates due to their feeding habits, ubiquitous distribution throughout many habitats and soil conditions, and importance in providing a food base for many small- to medium-sized predators. A diet of detritus, microflora, and microfauna, combined with direct contact with the surrounding soil, presents a potential link between soil contaminants and soil-invertebrate consumers. In addition. 30 USEPA 6899 000201 000 l !RFW 06` Breeding territories are established by male robins. Most foraging occurs close to these territories during the breeding season; however, ifdensities of robins are high in a given area or if food resources are limited, adult robins will leave to temporarily forage elsewhere. Outside of the breeding period, robins typically return to the same foraging sites and roost within 1to 3 kilometers (km) of these areas (U.S. EPA 1993). Exposure Profile Adult American robins are reported to weigh from 77.3 to 133.8 g (U.S. EPA 1993). Territory sizes vary from 0.3 to 1acre, with foraging home ranges reported up to 2 acres (U.S. EPA 1993). The lowest reported body weight (77.3 g) and the smallest reported home range of (0.3 acres) were assumed for this risk assessment. A food ingestion rate of 0.89 to 1.52 g/g BW/day and a water ingestion rate of 0.14 g/g BW/day are reported for this species (U.S. EPA 1993). Assuming a 77.3 g body weight, an American robin can be expected to consume 117.5 g/day of food and 10.8 g/day of water. Tne diet ofthe American robin consists of seasonally variable proportions of invertebrates (e.g., earthworms, snails, beetles, caterpillars, spiders) and fruit (e.g., dogwood, cherry, sumac, hackberries, raspberries) (U.S. EPA 1993, Ehrlich et al. 1988). During spring, summer, and fall, the dietary composition is reported to change from 93 percent invertebrates and 7 percent fruit in the spring (nesting season) to 92 percent fruit and 8 percent invertebrates in fall (migratory season). The summer dietary proportion is reported as 68 percent fruit and 32 percent invertebrates (U.S. EPA 1993). For the purposes of this risk assessment, a diet of 100%earthworms will be assumed. An incidental soil ingestion rate for the American robin could not be found in the literature. However, a soil ingestion rate of 10.4 percent of the diet reported for the American woodcock will be used as a substitute ingestion rate for the American robin (Beyer et al. 1994). Assuming a food ingestion rate of 117.5 g/day, the soil ingestion rate for the American robin is 12.2 g/day. .9.5 Red-tailed Hawk (Buieojamaciensis) as Representative of Carnivorous Birds. Justification. The red-tailed hawk was selected as representative of a carnivorous bird due to its dietary composition, relative abundant distribution, and likelihood of occurrence at the Dry Run Creek site. Its diet allows for the evaluation of contamination in site soils. In addition, the concentration ofcontaminants found in small mammal tissue will also provide an accurate dose to the red-tailed hawk which allows for the evaluation of contaminants in the food source. L ife H isto ry Red-tailed hawks are the most common and widespread American Buteo (Bull and Farrand 1977). Their habitat is highly variable, but they are commonly found in wooded areas near open land. They also inhabit plains, prairie groves, and deserts in the western United States (N'GS 1987). This species is absent, however, from tundra, and rare in extensive unbroken forest. An opportunistic feeder, the red-tailed hawk hunts from a perch or.on the wing for 33 U S E P A 6 9 0 2 0 0 9 000202 I IQC\A/ The fathead minnow, P. promelas, is widely distributed in North America and is found in a variety of habitats such as small streams, ponds, and small lakes. It is uncommon or absent in streams of moderate and high gradients. It is tolerant of high temperature' high turbidity, and low oxygen concenmations (U.S. EPA 1985). The fathead minnow is primarily omnivorous. Young typically feed on detritus, algae, and zooplankton. Adults feed on aquatic insects, worms, small crustaceans, and other animals. This species is consideredan important foodsource for other fish and birds (U.S. EPA 1985). Adult fathead minnows spawn in the spring and continue to spawn throughout most of the summer. The minimum spawning temperature appears to be approximately I6aC. The ovaries ofthe females contain eggs inall stages ofdevelopment, and they spawn repeatedly as the eggs mature. The average number ofeggs per spawn per female is 100 to 150. Larger females may lay 400 to 500 eggs per spawn. Hatching times depend on temperature and average about six days. In warm water with an ample food supply, spawning may occur as early as the first year. In cooler water with a moderate food supply, spawning usually occurs during the second year. Survival to the third year is relatively uncommon (U.S. EPA 1985). Exposure Profile Since direct contact with contaminated water in the toxicity evaluation is the primary route of exposure for fathead minnows in this risk assessment, the results of the test will be used to indicate exposure. 9.4 American Robin (Turdus migratorius) as Representative of Worm-eating Birds Justification The American robin was selected as representative of omnivorous and carnivorous birds because of its ubiquitous distribution and dietary composition. The preference for soil invertebrates in its omnivorous diet allows this species to be used as both an omnivorous and carnivorous receptor in this risk assessment. This species is also likely to occur at the Dry Run Creek site. Life History The American robin (Turdus migratorius) occurs throughout most ofthe continental U.S. and Canada, wintering inthe southern halfofthe U.S., Mexico, and Central America. Given the increase in open habitat and lawns, the robin's breeding range has expanded in the recent times. Habitat requirements for breeding robins include access to fresh water, protected nesting sites, and productive foraging areas. These requirements are commonly met in moist forests, swamps, open woodlands, andother open areas. Non-breeding robins occupy similar habitats although proximity to fruit bearing trees is of more importance. The primary foraging technique for robins is to hop along the ground in search of grounddwelling invertebrates, although they commonly search for insects and fruit in tree branches as well. The robin's diet during the breeding season consists mainly of invertebrates and some fruit, but fruit is the primary food consumed outside ofthe breeding season. As robins exhibit a low digestive efficiency for fruit, they often consume more than their own body weight in fruit to meet their metabolic needs (U.S. EPA 1993). 32 USEPA 6901 000203 USFW 0619 dose to the red fox which allows for the evaluation of contaminants in the food source. Life History Red fox inhabit open meadows, ditch banks, field and wood edges, fencerows, stream and lake borders, and farmlands (Hoffineister 1989; Jones and Bimey 1988; Merritt 1987). With the exception of the breeding season, red fox have no permanent home but sleep on the ground (Schwartzand Schwartz 1981). Aden, usually modified froman existing woodchuck or fox den, is dug during the breeding season and exceptionally cold winters (Barbour and Davis 1974). These scent-marked dens have multiple rooms, entrances, and trails leading to and from hunting areas (Schwartz and Schwartz 1981). In addition to their dens, both males and females will defend their scent-marked hunting territory from intruders (Jones and Bimey 1988). The red fox is primarily an opportunistic carnivore, consuming food items such as rabbits, opossums, muskrats, skunks, rodents, birds, eggs, carrion, invertebrates, snakes, and frogs (Barbour and Davis 1974; Merritt 1987). Some vegetable matter such as fruits and nuts are also consumed when in season (Jones and Bimey 1988). During times of abundant food supply, the red fox will bury surplus food to return to for consumption at a later time (Schwartz and Schwartz 1981). Male and female foxes pair for life, remaining together from midwinter to summer. Females bear one litter per year usually between March and April (Merritt 1987). Gestation periods last from about 49 to 56 days, with most averaging 53 days (Schwartz and Schwartz 1981). The pups are weaned at about 60 days, leave the den in the autumn, and are sexually mature by their first winter (Merritt 1987). Natural predators ofthe red fox are few but include large hawks and owls, and possibly coyotes (Merritt 1987; Schwartz and Schwartz 1981). Red fox may live from six to ten years in the wild (Schwartz and Schwartz 1981). Exposure Profile Adult red fox weigh from 2.7 to 7 kg (Barbour and Davis 1974; Jones and Bimey 1988). Home ranges vary from 245 to 1,235 acres (Merritt 1987). The food ingestion rates ofthe red fox range from0.069 g/g BW/day for a nonbreeding adult, to 0.16 g/g BW/day for a juvenile (U.S. EPA 1993). The water ingestion rate for an adult red fox is estimated to be approximately 0.086 g'g BW/day (U.S. EPA 1993). To express these values in units of g/day, the highest reported food ingestion rate of0.16 g/g BW/day and the water ingestion rate of 0.086 g/g BW/day were multiplied by the lowest reported body weight of 2.7 kg (2,700 g) to yield a food ingestion rate of 432 g/day and a water ingestion rate of232.2 g'day (232.2 mL/day). Forthe purposes of this risk assessment, a diet of 100% small mammals will be assumed. A soil ingestion rate of 2.8 percent ofthe total diet has been reported (Beyer et al. 1994) for the red fox. To express this value in units of g/day, the soil ingestion rate of 2.8 percent was multiplied by the food ingestion rate of432 g/day to yield a soil ingestion rate of 12.1 g/day. 2.9.7 Mink (Mustela vison) as Representative of Carnivorous Mammals Justification USEPA 6904 35 0 0 0 A 000204 USFW 0622 food items such as small mammals (e.g., mice, chipmunks, rabbits), birds (usually grounddwelling species), reptiles, insects, and occasionally, prey species that are too heavy to lift off the ground (Burton 1989). Tne breeding season starts with aerial courtship displays, commonly followed by mating on a perch and nest-building by both sexes. Nests are placed in tall trees, high rock ledges, or tall cacti and are often refurbished annually for use in consecutive years. Incubation of two to three eggs is carried out by both sexes and lasts for approximately30 days. The young are able to feed themselves at 4 to 5 weeks and fledge in about 45 days (Bull and Farrand 1977; Burton 1989). Exposure Profile Adult male and female red-tailed hawks are reported to weigh 960 g and 1,235 g, respectively (DeGraafand Rudis 1983; U.S. EPA 1993). Home ranges vary from 148.26 to 395.36 acres (Kirkwood 1980). The lowest reported body weight of 0.960 kg and the smallest reported home range of 148.26 acres were assumed for this risk assessment. The diet of a red-tailed hawk consists ofmammals, birds, reptiles, and insects which vary in importance with season and availability (U.S. EPA 1993). For the purposes of this risk assessment, the hawk will be assumed to consume 100%small mammals. Food ingestion rates are reported to range from 136 to 400 g/day (Kirkwood 1980). The highest reported food ingestion rate of400 g/day was assumed for this risk assessment. Awater ingestion rat of approximately 0.059 g/g BW/day has been estimated for this species (U.S. EPA 1993). To express this value in units ofg/day, the water ingestion rate was multiplied by the lowest reported body weight of960 g toyield awater ingestion rate of56.64 g/day (56.64 mL/day). A soil ingestion rate for the red-tailed hawk could not be found in the literature; therefore, the amount of soil predicted to be entrained in the digestive tract of a white-footed mouse was used to calculate this value. A soil ingestion rate of less than 2 percent of the total diet has been reported (Beyer et al. 1994) for the white-footed'mouse. From this value, a conservative soil ingestion rate of 1.9 percent of the total diet was assumed for the whi.tefooted mouse. To express this value in units of g/day, the soil ingestion rate of 1.9 percent was multiplied by the food ingestion rate ofthe white-footed mouse (4.50 gMay) (U.S. EPA 1993) to yield a soil ingestion rate of 0.09 g/day. This value was assumed to represent the amount of soil entrained in the digestive tract of the white-footed mouse that remains constant over time. To express 0.09 g in units of grams of soil per gram of mouse body weight, this value was divided by the lowest reported bodyweight (13 g) ofthe white-footed mouse (Merritt 1987) to yield a value of 0.007 g/g BW. This value was then multiplied by the food ingestion rate of the red-tailed hawk (400 g'day) to yield a soil ingestion rate of 2.8 g'day. 2.9.6 Red Fox (Vu!pes vuipes) as Representative of Carnivorous Mammals Justification The red fox was selected as representative of a carnivorous mammal due to its dietary composition, relative abundant distribution, and likelihood of occurrence at the Dry Run Creek site. Its diet allows for the evaluation ofcontamination in site soils. In addition, the concentration of contaminants found in small mammal tissue will also provide an accurate 34 USEPA 6903 goo. 000205 USFW 062- The mink was selected as representative of a carnivorous mammal due to its dietary composition, relative abundant distribution, and likelihood of occurrence at the Dry Run Creek site. Its diet allows for the evaluation of contamination in site soils. In addition, the concentration of contaminants found in clams and fish tissue will also provide an accurate dose to the mink which allows for the evaluation of contaminants in the food source. Lift History Mink are distributed over much of boreal North America, southward throughout the eastern United States and in the west to California, New Mexico, and Texas (Jones and Bimey 1988). They can be found in virtually any habitat containing permanent water thus, they are not commonly found inupland areas (Jones and Bimey 1988). Although primarilynocturnal, their activity often extends into midday (Hoffmeister 1989). Dens are always near water, and they are usually an old muskrat burrow or constructed by the mink itself(Jones and Bimey 1988). Males tend to live in their own burrows which are less elaborate than ones occupied by females (Barbour and Davis 1974). Home ranges tend to be linear since mink often follow a shoreline (Jones and Bimey 1988). Mink are solitary and mark their territories by spraying (Merritt 1987). Seasonal food availability governs the dietary composition (Barbour and Davis 1974). Their diets may consist of crayfish, frogs, fish, snakes, rodents, rabbits, and plants among other items (Jones and Bimey 1988; Schwartz and Schwartz 1981). Crayfish are a majorportion of the summer diet in many regions ofNorth America (Barbour and Davis 1981; Jones and Bimey 1988; Merritt 1987). Breeding occurs from January to early April with highly variable gestation periods ranging from 40 to 75 days (Merritt 1987; Schwartz and Schwartz 1981). A highly variable single liner of 1to 17 young may be produced (Schwartz and Schwartz 1981). Average liner sizes vary among regions (Barbour and Davis 1974; Hoffmeister 1989; Jones and Bimey 1988; Merritt 1987; Schwartz and Schwartz 1981). Young are weaned at about five to six weeks of age and are sexually mature by ten months (Merrin 1987; Schwartz and Schwartz 1981). Occasionally great homed owls, foxes, coyotes, bobcats, and dogs will prey on mink (Merrin 1987; Schwartz and Schwartz 1981). Although some individuals have lived up to six years, mink seldom exceed two years of age in the wild (Schwartz and Schwartz 1981). Effects Profile Adult mink weigh from 520 to 1,730 g (Merrin 1987; U.S. EPA 1993). Home ranges'vary from 19 to 1,900 acres (U.S. EPA 1993). A year-round food ingestion rate of 0.22 g/g BW/day has been estimated for both male and female mink (U.S. EPA 1993). To express this value in units of g/day, the food ingestion rate was multiplied by the lowest reported body weight (520 g) to yield a food ingestion rate of 114 g/day. An estimated water ingestion rate of 0.11g/g BW/day was reported for farmraised females (U.S. EPA 1993). To express this value in units of g/day, this water ingestion rate was multiplied by the lowest reported body weight of 520 g to yield a water ingestion rate of 57.2 g/day (57.2 mL/day). For the purposes of this risk assessment, a diet of 100% fish will be assumed. USEPA 6905 36 0 0 9 15 000206 USFW 0623 An incidental sediment ingestion rate was not available from the literature; therefore, a predicted incidental ingestion rate for sediment that maybe entrained in the digestive system of the prey item(fish) was used for this risk assessment. Consumption of this prey item was assumed to be the primarymechanism by which mink may incidentally ingest sediment. The derivation ofthe predicted level of incidental sediment ingestion via consumption of fish is described next. Life history information for the bluegill (Lepomis machrochirus) was used to predict the amount of sediment that may be ingested by mink via consumption of fish. Adult bluegills range in size from 100 to 230 mm (Pflieger 1975; Smith 1985). In keeping with the conservative approachofthis risk assessment, the amount ofsediment entrained in the lowest body size of 100 mm in length was predicted. The weight of a 100 mm bluegill was calculated to be 18.11 g based on the following algorithmrelating length to weight (Hillman 1982); log Weight (g) - -5.374 +3.316 log Length (mm) A daily food ingestion rate of 1.75 percent BW/day has been reported for the bluegill (Kolehmainen 1974). This provides a predicted intake rate of 0.32 g of food per day for a 18.11 g fish. A study evaluating the stomach contents of 153 bluegills reported an average content ofdetritus and sediment to be 9.6 percent of the total diet (Kolehmainen 1974). If a conservative assumption is made that 9.6 percent ofthe food ingested is entirely sediment,' it can be predicted that a fish of this size may contain 0.03 g of sediment in its digestive system. For the purpose of this model, it was assumed that the level of sediment contained in the digestive systemofa fish remains constant over time. This value (0.03 g) was divided by the predicted fish body weight (18.11 g) to express sediment entrained in fish digestive systems in units of grams of sediment per gram of fish body weight. This provided a value of0.0017 g sediment/g body weight. When this value is multiplied by the food ingestion rate of the mink (114 g/day), the predicted sediment ingestion rate for the mink through consumption of fish is 0.2 g/day. 2.9.8 Raccoon (Procyon lotor) as Representative of Omnivorous Mammals Justification The raccoon was selected as representative of a omnivorous mammal due to its dietary composition, relative abundant distribution, and likelihood of occurrence at the Dry Run Creek site. Its diet allows for the evaluation ofcontamination in site sediment. In addition, the concentration of contaminants found in forage fish tissue and clams will also provide an accurate dose to the raccoon which allows for the evaluation of contaminants in the food source. Life History Raccoons are medium-sized omnivores and are abundant throughout North America. Raccoons prefer aquatic habitats, particularly hardwood swamps, flood plains, freshwater wetlands, and salt marshes (Kaufrnann 1982). Raccoons have also adapted well to residential areas and farmlands. Raccoons rely heavily on surface waters for foraging and as a source 37 U S E P A 6 9 0 6 000207 I tOCAA/ n c o / i ofdrinking water (Srnewer 1943). Raccoons are active primarily fromduskto dawn (Stuewer 1943) but will alter their activities to opportunistically feed on whatever is available (Sanderson 1987). Forexample, raccoons living near a salt marsh may become active during the day to take advantage of feeding opportunities during low tide (Ivey 1948). Raccoons feed primarily on fruits, nuts, acoms, grains, insects, frogs, crayfish, and eggs (Palmer and Fowler 1975). Raccoons inthe southern regions ofthe United States are active year round (Goldman 1950). Adult raccoons are normally solitary but will come together for short periods oftime during mating (Kaufman 1982). Mating occurs fromMarch to June insouthern areas and each male may mate with several females during eachseason (Sanderson 1987; Kaufman 1982). Young males are normally not sexually mature in the first breeding season but mature later in the summer, while females mature in the first year (Sanderson 1951). The home range of a raccoon depends on the animal's age, habitat, food resources, and season (Sanderson 1987). Home ranges are typically a few hundred hectares (ha) but ranges as large as a few thousand ha have been reported (Sanderson 1987). Population densities also depend stronglyon the amount ofresources inthe area. Numbers of 0.1 to 0.2 animals per ha are common (Hoffman and Gottschang 1977). Raccoons are found near every aquatic habitat. During the last 50 years raccoon populations have increased greatly (Sanderson 1987). In Alabama, adult male raccoons weighed up to 8.8 kilograms (kg) (mean43 1kg) while adult females can weigh up to 5.9 kg (mean 3.67 kg) (Johnson 1970). Adult raccoons weigh between 2 and 12 kg (Nowak 1991), and consume 0.5 kg of food per day (Newell et al. 1987). Raccoons feed primarily on fruits, nuts, acoms, grains, insects, frogs, crayfish, eggs (Palmer and Fowler 1975). In a Maryland forested bottom land, the dietary composition of raccoons during the summer was principly made up of insects (39 percent), wild cherry (17 percent), blackberries (16 percent), crayfish (8 percent), snails (5 percent), herptiles (5 percent), fish (2 percent), rodents (2 percent), com (1 percent), and trace amounts ofSmilax, acoms and pokeberry (Llewellyn and Uhier 1952). At Washington state tidewater area raccoons displayed the following dietary composition: molluscs, mussels and oyster (44 percent), Crustacea, shrimp and crabs (25 percent), fish (9 percent), marine worms (20 percent), and Echiurida worms (1 percent) (Tyson 1950). The home range of a raccoon depends on the animal's age, habitat, food resources, and season (Sanderson 1987). Home ranges are typically a few hundred hectares but ranges as large as a fewthousand hectares have been reported (Sanderson 1987). The home range for adult male raccoon found in coastal Georgia raccoons is approximately 65 ha ( 18 SE) while the home range for adult females in the same area is approximately 39 ha ( 16 SE) (Lotze 1979). Population densities also depend strongly on the amount of resources in the area. Numbers of0.1 to02 animals per hectare iscommon (Hoffman and Gottschang 1977). EiSPOSUr? Profile For the purposes of this risk assessment, a body weight of 2 kg, an ingestion rate of 0.5 kg'day, and a diet of 80 percent forage fish and 20 percent clams were assumed. A soil ingestion rate of 9.4 percent of the diet has been reported for raccoons (Beyer et al. 1991). Multiplying the ingestion rate by 9.4 percent yields a sediment ingestion rate of0.047 kg'day. 38 U SE PA 6 9 0 7 ooo : 000208 USFW 0625 Adaily water ingestion rate of0.18 Liters per day (IVday) was calculated using an allometric equation derived by Calder and Braun (1983). A diet of 100% fish will be assumed. ^rr.raiied Shrew (Blarina brevicauda) as Representative of Insectivorous Mammals Justification The short-tailed shrew was selected as representative of insectivorous mammals because of its dietary composition, relative abundant distribution in both moist and dry habitats, and likelihood of occurrence at the Dry Run Creek site. Although their diets may consist of plants as well as insects, they tend to favor soil invertebrates when they are in abundance. Hence, by assuming that their dietary composition comprises solely invertebrates in this risk assessment, this species may represent an insectivorous mammal. Life History The short-tailed shrew is an extremely active, large, and heavy-bodied shrew common within its range (Jones and Bimey 1988). It occupies a variety of moist and dry habitats such as marshes, bogs, moist forest floors with ample decaying matter, brushland, fencerows, weedfields, and pastures (Barbour and Davis 1974; Jones and Bimey 1988). Short-tailed shrews are active both day and night throughout the year, although most of this activity is subnivean (Merritt 1987). During harsh winters, this species may undergo a period oftorpor' (Hoffmeister 1989). The home range ofthis species varies with their dramatic population cycles. In peak years, animal density may be greater than 25 individuals per acre (Schwartz and Schwartz 1981). In other years, this species may have an animal density of one individual per acre (Merritt 1987). Although short-tailed shrews strongly prefer animal matter, they'are opportunistic omnivores and will voraciously consume whatever food items are in ample supply (Barbour and Davis 1974). These food items include earthworms, slugs, snails, insects, arthropods, fungi, vegetable matter, seeds, snakes, salamanders, small mammals, and young birds CBarbour and Davis 1974; Jones and Bimey 1988; Schwartz and Schwartz 1981). Plant matter is generally consumed to a greater extent in winter (Schwartz and Schwartz 1981). In some regions, plant matter may constitute up to 20 percent of the shrew's diet (Barbour and Davis 1974). Submaxillary glands ptoduce a venom that quickly immobilizes their prey (Merritt 1987). Prey items that are not consumed immediately are stored in a cache (Merritt 1987). Using echolocation and scent-marking, short-tailed shrew rely heavily on their hearing and sense of smell to locate food and to move about (Hoffmeister 1989). An elaborate system ofrunways and tunnels are constructed usuallyjust a few inches below the ground surface (Schwartz and Schwartz 1981). Two types ofnests are built by this species, a breeding nest and a resting nest. Both nests are built underground beneath a log. rock, or other cover, and have multiple entrances. The breeding nest is typically larger than the resting nest (Merritt 1987). Breeding appears to commence in early spring and extend into the fall, although in some regions, breeding may subside in early and midsummer but peak again in early fall (Hofimeister 1989; Jones and Bimey 1988). Gestation periods are approximately 21 to 22 39 USEPA 6908 000:* 8 000209 USFW ncoo days with litter sizes ofapproximately four to ten young (Jones and Bimey 1988; Schwartz and Schwartz 1981). The young are fully mature from one to three months of age (Barbour and Davis 1974; Schwartz and Schwartz 1981). Both sexes may breed their first spring (Schwartz and Schwartz 1981). Natural predators of the short-tailed shrew include fish, snakes, owls, hawks, shrikes, opossums, raccoons, foxes, weasels, bobcats, skunks, and feral cats, although most of these predators do not consume the shrew (or at least all ofthe shrew) because of their distasteful musk glands (Barbour and Davis 1974; Jones and Bimey 1988; Merritt 1987; Schwartz and Schwartz 1981). The life expectancy ofa short-tailed shrew in the wild is approximately one year (Schwartz and Schwartz 1981). Exposure Profile Adult short-tailed shrews weigh from 12 to 30 grams (g) (Jones and Bimey 1988; Merrin 1987). Home ranges vary from0.5 to 1acre (Memtt 1987). Therefore, it was assumed that a short-tailed shrew could obtain 100 percent of its diet fromthe contaminated area (area use factor of 1), since the area comprising the on-site sampling locations was approximately 20 acres. Food ingestion rates ranging from 0.49 to 0.62 gram per gram of body weight per day (g/g BW/dav ) have been reported (U.S. EPA 1993). An average food ingestion rate of 7.95 g'dav has also been reported (U.S. EPA 1993). To express the former food ingestion rates in units of g/day for comparison to the latter ingestion rate, the former ingestion rates were multiplied by the lowest reported body weight of 12 grams to yield food ingestion rates of 5.SS to 7.44 g/day. Ofthese values, the highest food ingestion rate of7.95 g'dav will be used for the purposes of this risk assessment. Awater ingestion rate of0.223 g/g BW/day has been reported (U.S. EPA 1993). To express this value in units of g/day, the water ingestion rate was multiplied by the lowest reported body weight of 12 g to yield a water ingestion rate of 2.7 g'day (2.7 milliliters per day [mL/day]). Asoil ingestion rate for the short-tailed shrewwas not available from the literature, therefore, the soil ingestion rate ofthe opossum was used. The opossum's diet is similar to that of the short-tailed shrew since they are both opportunistic omnivores with a strong preference for animal matter (Schwartz and Schwartz 1981). A soil ingestion rate of 9.4 percent of the diet was reported for the opossum (Beyer et al. 1994). This value was multiplied by the highest food ingestion rate of the short-tailed shrew (7.95 g/day) to yield a soil ingestion rate of0.74 g'dav. For the purposes of the food chain model in this risk assessment, it was assumed that 100 percent of the diet of the short-tailed shrew was comprised of earthworms. .9.10 Meadow Vole (Microtuspennsylvanicus) as Representative of Herbivorous Mammals Justification The meadow vole was selected as representative of herbivorous mammals because of its dietary composition, abundance in North America, preference for moist areas, and likelihood of occurrence at the Dry Run Creek site. O O O -JS USEPA 6909 000210 USFW 0627 t.ife History The meadow vole is one ofthe largest and most abundant voles in North America (Jones and Bimey 1988; Merritt 1987). Although they are more commonly found in habitats such as moist meadows, bogs, swamps, stream banks, and lakeshores, they have also been known to inhabit cultivated fields, roadside ditches, and fencerows (Barbour and Davis 1974; Jones and Bimey 1988; Merritt 1987; Schwartz and Schwartz 1981). Dense vegetative cover appears to be one of the major prerequisites for habitation (Hoffineister 1989; Jones and Bimey 1988). The home range of the meadow vole varies in size with season, habitat, and population size (Jones and Bimey 1988). Populations tend to fluctuate drastically every two to five years, with peak population density levels exceeding 100 voles per acre (Barbour and Davis 1974; Jones and Bimey 1988). Activity occurs during both day and night, and throughout the year, although it is greatest at dawn and dusk (Barbour and Davis 1974). Well-worn intersecting runways under vegetative cover are distinctive of meadow vole inhabitation (Jones and Bimey 1988). Elaborate spherical nests are commonly built aboveground in the center of a tussock ofgrass, although underground nests are also built in drier areas (Barbour and Davis 1974; Jones and Bimey 1988). The meadow vole is herbivorous, feeding primarily on grasses, sedges, legumes, tubers, and roots (Merritt 1987); however, insectivory and cannibalism have been reported in some individuals (Barbour and Davis 1974; Hoffineister 1989). Bluegrass (Poa sp.) is a major component of the diet in some regions (Jones and Bimey 1988; Hoffineister 1989). This species hoards food for the winter in above- and below-ground caches (Merritt 1987). The meadow vole is one of the most prolific mammals, producing litter after litter in rapid succession (Barbour and Davis 1974). Breeding occurs during the warmer months of the year (Jones and Bimey 1988). The gestation period is about 21 days with litter sizes ranging from 1to 11 young (averaging four to seven) (Barbour and Davis 1974; Jones and Bimey 1988). The helpless young mature rapidly and may breed by 25 days of age (Barbour and Davis 1974). Meadow voles are preyed upon by nearly all species of predatory birds and mammals (Barbour and Davis 1974). These predators include owls, hawks, shrikes, bluejays, crows, foxes, weasels, mink, cats, raccoons, skunks, opossums, shrews, and snakes (Barbour and Davis 1974; Merritt 1987). Due to heavy predation, only a small proportion of the population exceeds sixty days of age (Schwartz and Schwartz 1981). Exposure Profile Adult meadow voles weigh from 20 to 65 grams (Merritt 1987; U.S. EPA 1993). The home range ofthis species varies from less than one acre to 3.2 acres (Merritt 1987). Therefore, it was assumed that a meadow vole could obtain 100 percent of its diet from the contaminated area (area use factor of 1), since the area comprising the on-site sampling locations was approximately 20 acres. A food ingestion rate ranging from 0.30 to 0.35 g/g BW/day, and a mean water ingestion rate of 0.21 g;g BW/day is reported for this species (U.S. EPA 1993). To express these values in units of g'day, the highest reported food ingestion rate of 0.35 g'g BW/day and the water 4 1 U S E P A 6910 GGQ-.C.O 000211 USFW 0628 ingestion rate of 0.21 g/g BW/day were multiplied faythe lowest reported body weight of 20 g to yield a food ingestion rate of 7.0 g/day and a water ineestion rate of 42 g/dav (4 > ml/day). " 3v A soil ineestion rate of2.4 percent ofthe toul d i * ' - *--n re^-- u '.tSe>'er et al- 1994) for the meadow voie. 10 express this *iue in units of g/day, the soil ingestion rate of 2.4 percent was multiplied by the food ingestion rate of 7.0 g/day to vi*<>*- .u uigcsuon rate of 0.17 g/day. For the purposes of the food chain model in this risk assessment, it was assumed that 100 percent of the diet was comprised of plants. .0 ASSUMPTIONS This risk assessment evaluates exposure to contaminants through food and incidental sediment/soil ingestion. The following conservative assumptions were made to conduct this risk assessment in the absence of sitespecific data: The maximum ofthe contaminant levels measured in sediment, soil, or water collected on site was used in risk calculations. The maximum concentrations of COCs reported in sediment, soil, water, and biota were assumed to be present site-wide. .; An area use factor (AUF) of 1was assumed for all species using the site for feeding. Contaminants were assumed to be 100 percent bioavailable. Dietary composition information was obtained fromthe literature for the receptor species. However, simplifications of complex diets were performed for the receptors. A literature search was conducted to determine the chronic toxicity of the contaminants of concern when ingested by the indicator species. If no toxicity values could be located for the receptor species, values reported for a closely related species were used. All studies were critically reviewed to determine whether study design and methods were appropriate. When values for chronic toxicity were not available, LD,a (median lethal dose) values were used. For purposes of this risk assessment, a factor of 100 was used to convert the reported LDJ0 to a No Observed Apparent Effect Level . (NOAEL). A factor of 10 was used to convert a reported Lowest Observed Adverse Effect Level (LOAEL) to a NOAEL. and a factor of 10 was used to convert a reported LD,, to a LOAEL. Ifseveral toxicity values were reported for a receptor species, the most conservative value was used in the risk calculations regardless of toxic mechanism. Toxicity values obtained from long-term feeding studies were used in preference to those obtained from single dose oral studies. No other safety factors were incorporated into this risk assessment. In some cases, contaminant doses were reported as part per million contaminant in diet. These were convened to daily intake (in milligrams per kilogram body weight per day; mg/kg-day) by using the formula: Intake (mg'kg/day)*Contaminant Dose (mg/kg diet) x Ingestion Rate (kg/day) x 1/Bodyweight (kg) 42 0 0 0-: n USEPA 6911 000212 USFW 0629 This conversion allows dietary toxicity levels cited for one species to be convened to a daily dose for a different species based on body weight. For the purposes of this risk assessment, incidental nii/sediment ingestion was also included in the calculation to determine the total daily intake for the recepto. .r -n-;- j,;ivdose may then be used to evaluate the risk to other species if no specific toxicity data art available for a (ccepiu>. Some containin ofcericer" (e.g. aluminum) are not food chain accumulators, but instead are direct toxins when ingested at the prescn-.! levels. EFFECTS PROFILE Many contaminants detected at the Dry Run Creek site do not have benchmarks. This excluded them from further consideration in this risk assessment, but does not exclude them as potential contaminants of concern. Based on the results ofthe preliminary risk assessment, the following compounds were considered COCs and theirtoxic effects are presentednext: fluoride, trichlorofluoromethane, aluminum, arsenic, beryllium, chromium, copper, iron, lead, manganese, nickel, vanadium, and zinc. Based on the chemistry results, these compounds will be further evaluated using food chain accumulation models. Contaminants exceeding their respective benchmarks are assumed to be affecting receptor species and negatively impacting species, populations, and communities in the aquatic and terrestrial ecosystems at the Dry Run Creek site. 4.1 Fluoride Maurer et al (1990) identified skeletal and dental abnormalities in rats that were exposed to sodium fluoride for a period of 99 weeks. The LOAEL identified in this study was 4 mg Fl/kg BW/day. A NOAEL was calculated from the LOAEL using an accepted conversion factor of 10. Based on these results, a LOAEL of 4mg/kg BW/day and an estimated NOAEL of 0.4/kg BW/dav will be used to evaluate the risk posed by fluoride mammalian receptors Fleming et al. (1987) found significant growth rate reduction in European starling fed a diet containing as low as 13 mg Fl/kg BW/day. No effects were observed at 10 mg Fl/kg BW/day. As such, this risk assessment will estimate fluoride related risk using a LOAEL of 13 mg'kg BW/day and a NOAEL of 10 mg'kg BW/day. 4.2 Organofluorides No studies pertaining to the dietary toxicity oftrichlorofluoromethane or any other fluorinated oreanic compound was found in the literature. 4.3 Aluminum Dixon et al. (1979) conducted a study that evaluated the reproductive success of rats exposed to aluminum in drinking water for 90 days prior to breeding. The highest dose administered was 77.5 milligrams per kilogram body weight per day (mg/kg BW/day) and did not result in reproductive abnormalities. Lai et al. (1993) conducted a 180-day drinking water study in which rats were exposed to 55 mg'kg BW/day of aluminum. At this dose, behavioral effects were observed, including a significant reduction in spontaneous locomotor activity and significant deficits in acquisition and retention of learned responses. Based on these results, a LOAEL of 55 mg/kg BW/day and an estimated NOAEL of 5.5 mg'kg BW/day will be used to evaluate the risk posed by aluminum to mammalian receptors (Table 1). USEPA 6912 43 000213 0 0 0 .;' '' IO C 1 M n o n , No effects were observed when Japanese quail were fed a diet containing 0.05 percent (84 mgkg BW/day) aluminum for four weeks (Hussein et al. 1988). When quail were fed a diet containing 0.1 percent (165 mg/kg BW/day) aluminum, a decrease in egg shell breaking strength was observed. Finally, when quail were feda diet containing 0.15 percent (257 mg/kg BW/day) aluminum, a decrease in body weight, egg shell strength, and egg shell production was observed. A48-day feeding study using chickens concluded that dietary levels of28.4 mg/kg BW/day aluminum resulted in a decrease in weight gain, feed intake, andplasma inorganic phosphorus, as well as an increase in plasma calcium (Hussein 1990). However, only the altered metabolismofcalciumand phosphorus could be attributed to the direct effects of aluminum. The associated NOAEL for this effect is 22.8 mgkg BW/day. Because a range of concentrations were used and the endpoints were ecologically significant and related to the dose, the study by Hussein et al. (1988) was used to the develop the NOAEL and LOAEL values. ANOAEL of 84 mg/kg BW/day and a LOAEL of 165 mg/kg BW/day will be used to evaluate the risk posed by aluminum to avian receptors (Table 1). 4.4 Arsenic Several studies were located which determined the effects ofAs to mammals. A study conducted on cats indicated that a chronic oral toxicity dose was 1.5 mg/kg BW/day (Pershagen and Vahter 1979). In addition. National Resources Council of Canada (1978) states that mammals in general have oral LDwSthat range from 10 to 50 mg/kg of lead arsenate. A study conducted on mice indicated an oral dose LDj0of39.4 mg/kg BW/day and an oral dose LD#of 10.4 mg/kg BW/day after 96 hours (NAS 1977). For the purposes of this risk assessment, the chronic value for the cat was used to calculate HQs for mammals (1.5 mg/kg BW/day). This value was convened to a NOAEL by dividing by a factor of 10. Eisler (1988a) reviewed several studies in which the toxicity of inorganic arsenicals were measured. Inorganic As is more mobile than organic As andmaypose greaterrisk by leaching into surface water. Studies were also described in which organoarsenical compounds were measured. Studies indicate that sensitive species include the California quail (single oral dose LDJ0of 47.6 mgkg BW/day) (Hudson etal. 1984) and chicken (single oral dose LD* of 33 mg/kg BW/day) (NAS 1977). For the purposes of this risk assessment, a value of 3.3 mg/kg BW/day was used to determine the HQ to birds. This value was convened to a NOAEL by dividing by a factor of 10. 4.5 Beryllium Two separate chronic dietary exposure studies using rats reponed similar musculoskeletal effects. Guyatt et al. (1933) fed large amounts of beryllium carbonate to rats at concentrations of 10, 20,40, 80, 160, and 240 mgkg BW/day. Rats from all exposure levels developed rickets, with the fragility of the bones varying directly with the exposure concentration. Similar results were reponed by Jacobson (1933) who reponed severely weakened bones in rats fed beryllium carbonate at dietary levels of 121 and 242 mgkg BW/day. For this risk assessment, a dietary exposure level of 10 mgkg BW/day was used to estimate risk of beryllium to the short-tailed shrew. ANOAEL of 0.10 mgkg BW/day was derived fromthis LOAEL using an accepted conversion factor of 10. No studies pertaining to the dietary toxicity of beryllium to avian receptors were found in the literature. 4.6 Chromium USEPA 6913 44 G00`:'-'53 000214 USFW 0631 Heinz and Haseltine (1981) exposed 2- to 3-year old breeding pairs of black ducks (Anas mbripes) to a diet containing 0, 20, or 200 mg/kg, wet weight, (0.2.77, or 27.77 mg/kg BW/day) of Cr*1as chromium potassium sulfate [CrK(SO^'PHjO] for a period of approximately five months, until the onset of egg-laying by the females. Hatched ducklings were then fed a mash diet containing the same Cr concentrations that the parents were fed. Seven-day old chicks were tested for avoidance behavior in response to a fright stimulus. None of the Cr concentrations resulted in alteration of avoidance behavior. However, Haseltine et al. (1985), in an unpublished study reported by Eisler (1986a) notes that black duck ducklings suffered reduced survival and altered growth patterns when exposed to 10 mg/kg and 50 mg/kg of an unspecified Cr'5compound in their diets. The percent reduction in survival and a detailed explanation of the altered growth patterns were not available in this unpublished study. For the purposes ofthis risk assessment, dietary levels of 10 mg/kg (1 mg/kg BW/day) of Cr in prey was used as a LOAEL for the avian species. However, due to the conflicting results, a NOAEL was derived from the same study in which the LOAEL was selected to maintain a degree of consistency regarding the Cr species evaluated. A NOAEL of0.1 mg/kg BV//day was derived from the LOAEL using a conversion factor of 10. A study conducted with dogs indicated that 2.5 mg/kg/day of Cr**ingested in the diet caused death (Steven et al. 1976). For the purposes of this risk assessment, a LOAEL of 0.25 and a NOAEL of 0.025 were used for the red fox, raccoon, and mink. 4.7 Copper One study was located which determined the effects of ingestion ofCu to mammalia species. An oral dose of 100mg/kg/dayto a dog caused death (OHMD 1987). For the purposes ofthis risk assessment, a LOAEL of 10 mg/kg/day was used and a NOAEL of I mg/kg/day were used for the exposure of mammals. Several studies were located which determined the effects of Cu on chickens. A dose of 350 mg/kg (61.3 mgkg/day) caused a significant decrease in growth and food consumption (Smith 1969). Another study found that a dose of 325 mg/kg (23.5 mg/kg/day) caused respiratory problems (Hatch 1978). Assuming that respiratory problems are an acute effect, a LOAEL of 2.35 mg/kg/day and a NOAEL of 0.235 mg/kg/day were used to determine risk to avian species. 4.8 Iron No studies pertaining to the dietary toxicity of iron to mammalian or avian receptors were found in the literature. 4.9 Lead The gastric motility of adult male and female red-tailed hawks fed 0.82 and 1.64 mg Pb/kg BW/day in a single oral dose was evaluated through the use of surgically implanted transducers for a period of three weeks following the dose. Neither concentration had any effect on gastric contractions or egestion of undigested material pellets (Lawler et al. 1991). A study conducted on red-tailed hawk found that 3 mg/kg/dav of Pb caused the clinical symptoms of Pb poisoning (Reiser and Temple 1981). A similar study found that 3 mg/kg'day fed to starlings caused a reduction in muscle condition and altered their feeding activity (Osbome et al. 1983). For USEPA 6914 45 O 0O -\c 000215 I ICCHA/ n c n r the purposes of this risk assessment, a LOAEL of 3 mg/kg/day was used to determine risk to avian species and a NOAEL of 0.3 was used. Several studies were located which determined the effects of Pb ingestion to mammals. A study conduced on mice indicated that 1.5 mg/kg/day ofPb caused a reduction in success of implanted ova (Clark 1979). Another study found that 22 mg/kg/day caused a reduction in the frequency of pregnancy when the dose was administered 3 to 5 days following mating (Clark 1979). For the purposes of this risk assessment, a NOAEL of0.15 mg/kg/day and a LOAEL of 1.5 mg/kg/day were used to determine risk to mammals. 4.10 Manganese The effects ievels for manganese toxicity vary widely, most likely attributable to the form of manganese tested. Rats exposed to 13 mg/kg BW/day of manganese as Mn304 in their diet for 224 days exhibited reduced testosterone levels (Laskey et al. 1982). In mice, a dietary level of 140 mg/kg BW/day, also of Mn304 for 90 days resulted in decreased activity (Gray and Laskey 1980). A much higher exposure concentration of2,300 mg/kg BW/day of manganese as MnC12 resulted in reduced dopamine levels (Gianutsos and Murray 1982). In contrast, levels as high as 930 mg/kg BW/day ofmanganese as MnS04 for 103 weeks had no efFec on the respiratory, cardiovascular, gastrointestinal, hematological, musculoskeletal, hepatic, renal, dermal, and ocular systems of mice (Hejtmancik et al. 1987). For this risk assessment, a dietary exposure level of 13 mg/kg BW/day will be used as a LOAEL to estimate risk ofmanganese tothe selectedmammalian receptor. ANOAEL of 1.3 mg-kg BW/day was derived from this LOAEL using an accepted conversion factor of 10. No studies pertaining to the dietary toxicity of manganese to an avian receptor were found in the literature. 4.11 Nickel Several studies were available which determined the effects ofNi ingestion to mammals. Wistar rats fed Ni sulfate indicated a NOAEL of 187.5 mg/kg/day to most systems except for body weight. This level of Ni sulfate caused a 27 to 29 percent decreased body weight (Ambrose et al. 1976). In a similar study with a beagle, a NOAEL of 62.5 mg/kg/day was noted (Ambrose et al. 1976). For the purposes ofthis risk assessment a NOAEL of62.5 mg/kg/day was used to determine risk to mammals. This value was converted to a LOAEL of 625.0 mg/kg'day by multiplying the NOAEL by a factor of 10. No studies were available that determined the dose ofNi to avian species. Tnerefore, the risk to avian species from ingested Ni will not be determined. 4.12 Vanadium Gavage studies in mice have found an LC50 of31 mg Vn/Kg diet (Schroeder and Balassa 1967). This dose was converted to a LOAEL of 3.1 mg/kg and a NOAEL of 0.31 using an accepted factor of 10 conversion. This food dose was convened to a daily dose by multiplying the LOAEL or NOAEL concentration by an ingestion rate commonly observed in mice (0.003 kg of food/day) and then by the inverse of the body weight (0.025 kg)(RTECS 1985). This calculation resulted in a LOAEL of 0.372 46 USEPA 6915 000'io.[) 00023.6 USFW 0632 mg V/kg BW/day and a NOAEL of 0.0372 mg V/kg BW/day. These values will be used to estimate risk to mammalian receptors in this risk assessment. Rosomer (1960) exposed chickens tovarying concentrations ofvanadium. The study involved feeding 4 replicates of 13 chickens varying concentrations of vanadium for a period of 21 days. The study found that a dietary level of40 mg/kg inthe diet resulted in a marked depression in weight gain and efficiency of food utilization. At levels of 200 mg/kg, mortality was noted in all test chickens. The authors reported that a dietary level of 20 mg/kg could be tolerated with no resultant toxic effects. This dietary level was converted to a daily dose as above by multiplying the dietary concentration by a representative chicken ingestion rate (0.140 kg/day) and then by the inverse of the body weight (0.800 kg)(RTECS 1985). This calculation resulted in LOAEL of 7 mg V/kg BW/day and a NOAEL of 3.5 mg V/kg BW/ day. These values will be used to estimate risk to avian receptors. 4.13 Zinc Several studies were available which determined the effects of ingested Zn to birds. A concentration of 144.5 mg/kg/day caused a decrease in growth and anemia in chickens (Stahl et al. 1989). In a similar study conducted on chickens, a concentration of 361 mg/kg/day caused a reduction in body weight (Dean et al. 1991). In a study conducted on Japanese quail, a concentration of 139 mg/kg/day caused 7 percent mortality in chicks and reduced food intake (Hill and Camardese 1986). For the purposes of this risk assessment, a LOAEL of 139 mg/kg/day was used to determine the effects to avian species. This value was convened to aNOAEL of 13.9 mg/kg/day by dividing the LOAEL by a factor of 10. A study conducted on dogs, indicated that 1,000 mg/kg (25 mg/kg/day) caused no effects after one year (NAS 1979). For the purposes of this risk assessment, a LOAEL of 250 and aNOAEL of 25 were used to determine risk to the fox and the mouse. In a study conducted on fenets, a dose of 370 mg'kg day caused a decrease in food intake and weight loss. Because the ferret is similar to the mink, a LOAEL of 370 mg/kg/day was used and a NOAEL of 37 was used to determine risk to the mink. RISK CHARACTERIZATION The following method was used to calculate risk. To estimate the risk to wildlife in the model systems utilizing the Dry Run Creek site, implications of the exposure concentrations need to be determined. The HQ method (U.S. EPA 1989, Bamthouse et al. 1986) compares exposure concentrations to ecological endpoints such as reproductive failure or reduced growth. The comparisons are expressed as ratios of potential intake values to population effect levels, or: Hazard Quotient (HQ) - ______Mean Exposure Concentration No Observed Adverse Effect Level (NOAEL) A HQ greater than one indicates that exposure to the contaminant has the potential to cause adverse effects in the organism. A HQ less than one does not indicate a lack ofrisk. The HQ should be interpreted based on the severity of the effect reported. The results of the risk characterization are presented next. 5.1 Benthic Invertebrate Community Structure and Function The benthic invertebrate community in Dry Run appears to be at risk for two reasons. The benthic community survey showed a decrease in community taxonomic diversity and abundance in Dry Run as compared to the Reference stream. Since land use and available habitat are the same along both 47 USEPA 6916 0 0 0 -lcS 00021V streams, the decrease in diversity and abundance in Dry Run may be attributed to contamination present in sediments in Dry Run. In addition, the amphipod toxicity test dearly demonstrates that acute exposure sub-lethal effects can be produced in the benthic community, especially under conditions present in Tributary A and in Area II. The observed negative growth effect was significantly negatively correlated with fluoride, aluminum, calcium, magnesium, nickel, potassium, and sodium. Further, there were strong negative associations between the growth endpoint and chromium, copper, lead, and zinc concentrations, although the relationships were not significant at the 0.10 level. Since the sediments closer the landfill along the whole Dry Run reach appear to be enriched with metals, the observed toxicity may represent a significant threat. 5.2 Soil Invertebrate CommunityStructure and Function The soil invertebrate community does not appearto be at risk based on current soil conditions at Dry Run. The earthworm toxicity test identified no problems with survival or growth. 5.3 Fish Communities The fish community at Dry Run may be at risk. Results ofthe fathead minnow toxicity bioassay show that water conditions in Upper Tributary A induce mortality to larval fish. This mortality could not directly be associated with a suite ofcontaminants as inthe amphipod test, but survival was negatively correlated with potassium concentrations, however this correlation was not statistically significant at the 0.10 level. There was a significant positive correlation between fathead survival and iron concentrations in the filtered water samples. Lowspecies diversity and abundance observed during the eiectroshocking effort may be reflected by the results of the toxicity test. 5.4 Worm-eating Birds Aconservative risk assessment model based on wet-weight concentrations ofcontaminants for the Dry Run Creek site has determined that worm-eating birds may be at risk due to ingestion ofcontaminated forage, soil, and water. The model predicts that aluminum, chromium, copper, lead, vanadium, zinc, and fluoride are risk factors based on conservative inputs. By default, beryllium, iron, manganese, nickel, and trichlorofluoromethane are risk factors due to lack of toxicological benchmarks for these compounds. Food chain risk calculations and resultant hazard quotients are presented in Table 42. 5.5 Carnivorous Birds Aconservative risk assessment model based on wet-weight concentrations ofcontaminants for the Dry Run Creek site has determined that carnivorous birds may be at risk due to ingestion of contaminated forage, soil, and water. The model predicts that aluminum, chromium, copper, lead, zinc, and fluoride are risk factors based on conservative inputs. By default, beryllium, iron, manganese, nickel, vanadium, and trichlorofluoromethane are risk factors due to lack of toxicological benchmarks for these compounds. Food chain risk calculations and resultant hazard quotients are presented in Table 42. 5.6 Carnivorous Mammals (Terrestrially feeding) Aconservative risk assessment model based on wet-weight concentrations of contaminants for the Dry Run Creek site has determined that carnivorous mammals may be at risk due to ingestion of contaminated forage, soil, and water. The model predicts that aluminum, chromium, copper, lead. 48 009 USEPA 6917 000218 USFW 0635 manganese, vanadium, and fluoride are risk factors based on conservative inputs. By default, iron and trichlorofluoromethane are risk factors due to lack oftoxicological benchmarks for these compounds. Food chain risk calculations and resultant hazard quotients are presented in Table 42. 5.7 Piscivorous Mammals Aconservative risk assessment model based on wet-weight concentrations ofcontaminants for the Dry Run Creek site has determined that piscivorous mammals may be at risk due to ingestion of contaminated forage, soil, and water. The model predicts that chromium, manganese, and fluoride are risk factors based on conservative inputs. Trichlorofluoromethane is not considered a risk factor because it was not detected in site sediments. Food chain risk calculations and resultant hazard quotients are presented in Table 42. 5.8 Omnivorous Mammals Aconservative riskassessment model based on wet-weight concentrations ofcontaminants for the Dry Run Creek site has determined that omnivorous mammals may be at risk due to ingestion of contaminated forage, soil, and water. The model predicts that arsenic, chromium, copper, manganese, vanadium, and fluoride are risk factors based on conservative inputs. Trichlorofluoromethane is not considered a risk factor because it was not detected insite sediments. Food chain risk calculations and resultant hazard quotients are presented in Table 42. 5.9 Insectivorous Mammals Aconservative risk assessment model based on wet-weight concentrations ofcontaminants for the Dry Run Creek site has determined that insectivorous mammals may be at risk due to ingestion of contaminated forage, soil, and water. The model predicts that aluminum, chromium, copper, lead, manganese, vanadium and fluoride are risk factors based on conservative inputs. By default, iron, and trichlorofluoromethane are considered risk factors due to lack of toxicological benchmarks for these compounds. Food chain risk calculations and resultant hazard quotients are presented in Table 42. 5.10 Herbivorous Mammals Aconservative risk assessment model based on wet-weight concentrations of contaminants for the Dry Run Creek site has determined that herbivorous mammals may be at risk due to ingestion of contaminated forage, soil, andwater. The model predicts that aluminum, chromium, lead, manganese, and fluoride are risk factors based on conservative inputs. By default, iron, vanadium, and trichlorofluoromethane are considered risk factors due to lack of toxicological benchmarks for this compound. Food chain risk calculations and resultant hazard quotients are presented in Table 42. 6.0 UNCERTAINTY ANALYSIS There are factors inherent in the risk assessment process which contribute to uncertainty and need to be considered when interpreting results. Major sources of uncertainty include natural variability, error, and insufficient knowledge. Error can be introduced by use ofinvalid assumptions inthe conceptual model. Conservative assumptions were made in light of the uncertainty associated with the risk assessment process. This was done to minimize the possibility of concluding that no risk is present when a threat actually does exist (e.g., elimination of false negatives). Whenever possible, risk calculations were based on conservative values. For example, NOAELs 49 U S E P A 6918 000219 i io n r r n c used to calculate HQs were the lowest values found in the literature, regardless of toxic mechanism. An important contributor to uncertainty is the incompleteness of the data or information upon which the risk assessment is based. Risk calculations are based on maximum COC levels in sediment, water, and soil samples. Literature values for the toxicity of COCs were not available for all receptor species. An attempt was made to identify studies using closely related species to make risk estimates for the selected receptors. Species respond differently to exposure to toxins; responses to COCs by the indicatorspecies may be different from species for which the toxicity data are reported. Methodological problems were also apparent inseveral ofthe studies from which NOAELs were obtained. Unfortunately, studies which were more suitable for this assessment were not found for some of the selected receptors. A literature search was conducted to identify appropriate NOAELs and LOAELs for this risk assessment. The values used to calculate HQs were the lowest values found in the literature. In many of the studies reviewed, adverse effects were observed at the lowest exposure concentration. This made it impossible to identify appropriate NOAELs for some receptors. In these cases, a factor of 10 was used to convert the LOAEL to a NOAEL, which adds uncertainty to the NOAEL-based calculations. Doses in toxicological studies can be reported in units of mg contaminant/kg diet, or in units of mg contaminant/kg body weight/day. All doses reported as mg/kg in diet were convened to units of mg/kg BW/day. Ifbody weights were reponed for the test animals ina givenstudy, these values were used for making this conversion. Otherwise, the body weight and ingestion rate for the species reponed in other literature sources were used. Another source of uncertainty arises fromthe use of toxicity values reponed in the literature which are derived from single-species, single-contaminant laboratory studies. Prediction of ecosystem effects from laboratory studies is difficult. Laboratory studies cannot take into account the effects ofenvironmental factors which may add to the effects of contaminant stress. NOAELs were generally selected from studies using single contaminant exposure scenarios. Species utilizing the Dry Run Creek site are exposed to a variety of contaminants. There is very little information available in the literature regarding the rates of incidental soil/sediment ingestion for wildlife species. In this risk assessment, most of these values were based on estimates reponed for species similar to the indicator species. Exposure concentrations were calculated for each target receptor species based on levels of contaminants detected in site media, daily food ingestion rates, incidental soil/sediment ingestion rates, and body weight reported in the literature. Tnis ecological risk assessment was conducted with the intent ofcompleting a baseline risk assessment. In this risk evaluation it is concluded that a "potential ecological risk"exists if the HQ calculated from the maximum area concentration and the NOAEL equals or exceeds one. Within the calculation spreadsheets, alternate calculations were made using LOAEL toxicity benchmarks. CONCLUSIONS 7.1 Benthic Invertebrate Community Structure and Function Data from both the benthic survey and the toxicity tests indicate that fluoride and metal contamination may be a significant problem in Dry Run. Numerous fish kills historically reported in Dry Run also 50 USEPA 6919 OOOAoO 000220 USFW 0637 provide evidence for potential effects on the benthic community. 7.2 Soil Invertebrate Community Structure and Function The structure and function of the soil invertebrate community does not appear to be at risk under current conditions found atthe Dry Run Creek site. However, since earthworms comprise a significant amount ofthe forage base ofsome organisms (e.g. American robins, short-tail shrews, etc.), food chain problems may result from contaminants being tied up in the earthworm tissue. Based on our food chain models, it appears that this may be the case. 7.3 Fish Communities It was shown through the results of the fathead minnow bioassay, that larval fish were susceptible to contamination currently present near the landfill outfall at Dry Run. This finding is further supported by the results of the benthic invertebrate toxicity tests, where toxicity was observed at the same location. Negative effects of contaminants on the benthic community may directly affect fish communities, in that a portion of the fish food base in Dry Run (i.e. benthic invertebrates) may also be removed from the system. Reports of historical fish kills are also an important piece of evidence that suggests an ecological risk. In addition, high levels of metals were noted in the fish, which may present problems to upper level consumers due to dietary toxicity. 7.4 Worm-eating Birds Results ofthe food chain model for worm-eating birds such as the American robin indicate a potential risk due to metals, fluoride, and trichlorofluoromethane. This risk is associated with these contaminants inthe soil and/or in earthworm tissue. Reports of historical wildlife kills also suggest ecological risk to avian receptors. 7.5 Carnivorous Birds Results ofthe food chain model for carnivorous birds such as the red-tail hawk indicate a potential risk due to metals, fluoride, and trichlorofluoromethane. This risk is associated with these contaminants in the soil and'or in small mammal tissue. Reports of historical wildlife kills also suggest ecological risk to avian receptors. 7.6 Carnivorous Mammals Results of the food chain model for terrestrially feeding carnivorous mammals such as the red fox indicate a potential risk due to metals, fluoride, and trichlorofluoromethane. "his risk is associated with these contaminants in the soil and/or in small mammal tissue. Reports of historical wildlife kills also suggest ecological risk to mammalian receptors. 7.7 Piscivorous Mammals Results of the food chain mode! for piscivorous mammals such as the mink indicate a potential risk due to metals, fluoride, and trichlorofluoromethane. This risk is associated with these contaminants in the soil and'or in fish tissue. Reports of historical wildlife kills also suggest ecological risk to mammalian receptors. 7.8 Omnivorous Mammals 51 USEPA 6920 000 ,so 000221 o 5. i i o c \ M n f i 8 8 Results ofthe food chain model for omnivorous mammals such as the raccoon indicate a potential risk due to metals, fluoride, and trichlorofluoromethane. This risk is associated with these contaminants in the soil and/or in fish tissue. Reports of historical wildlife kills also suggest ecological risk to mammalian receptors. 7.9 Insectivorous Mammals Results of the food chain model for insectivorous mammals such as the short-tail shrew indicate a potential risk due to metals, fluoride, and trichlorofluoromethane. This risk is associated with these contaminants inthe soil and/or in earthworm tissue. Physiological abnormalities, specifically the tooth structure ofthe shrews taken on site, further suggest ecological risk. In addition to the direct potential risk for the shrews, some of these animals had high concentrations of metals and fluoride in their tissues. This could present problems to organisms that feed on shrews and other small mammals on the site due to dietary toxicity. Reports of historical wildlife kills also suggest ecological risk to mammalian receptors. 7.10 Herbivorous Mammals Results of the food chain model for herbivorous mammals such as the meadow vole indicate a potential risk due to metals, fluoride, and trichlorofluoromethane. This risk is associated with these contaminants in the soil and/or in plant tissue. In addition to the direct potential risk for the voles; some of these animals had high concentrations of metals and fluoride in their tissues. This could present problems to organisms that feed on voles and other small mammals on the site due to dietary toxicity. Reports of historical wildlife kills also suggest ecological risk to mammalian receptors. SUMMARY During the past several years, a fanner who grazes his cattle along the reach of Dry Run Creek, has reported severe abnormalities in his herd. These abnormalities have included an increased incidence of stillborn calves, blindness in newborn and adult cattle, erratic behavior, stiffness of gait in adult cattle, abnormal posture, mottled teeth, and a high mortality rate across all age classes of his herd. In addition to problems with his herd, the farmer and others have also reported numerous fish kills in Dry Run, and wildlife kills (e.g. deer) for animals drinking from Dry Run. The results of this risk assessment support his assertion that effluent from the Dry Run Creek landfill may be having adverse effects on the ecological communities that inhabit the old field, deciduous forest, meadow, stream, and riparian habitats that are present on the site. These effects may be related to enriched levels of metals, fluoride, and trichlorofluoromethane that appear to be resultant of the landfill drainage. At a minimum, the symptoms manifest by the herd are characteristic of fluoride toxicity, and consistent with the conclusions of the risk assessment. In addition to the compounds that were studied in this risk assessment, numerous other compounds were present in Dry Run (specifically those identified as TICs or Tentatively Identified Compounds in the BNA scan) that could not be accurately identified. These compounds may also present a threat to the system, an certainly merit further investigation. The DuPont landfill that drains into Dry Run is the only apparent source of trichlorofluoromethane in soils adjacent to the stream. USEPA 6921 000222 1ISFW 0639 LITERATURE CITED Agency for Toxic Substances and Disease Registry (ATSDR). 1990. Toxicological Profilefor Aluminum. Prepared bv Sciences International Inc. Under U.S. Department of Health and Human Services Contract No. 205-93-0606. Research Triangle Park, NC. Agency for Toxic Substances and Disease Registry (ATSDR). 1990. Toxicological Profilefor Manganese. Prepared bv Sciences International Inc. Under U.S. Department of Health and Human Services Contract No. 205-93-0606. Research Triangle Park, NC. Azency for Toxic Substances and Disease Registry (ATSDR). 1991. Toxicological Profilefar Vanadium. Prepared by Sciences International Inc. Under U.S. Department of Health and Human Services Contract No. 205-93-0606. Research Triangle Park, NC. Agency for Toxic Substances and Disease Registry (ATSDR). 1993. Toxicological Profilefor Beryllium. Prepared by Sciences International Inc. Under U.S. Department of Health and Human Services Contract No. 205-93-0606. Research Triangle Park, NC. Agency for Toxic Substances and Disease Registry (ATSDR). 1996. Toxicological Profilefor Nickel. Prepared by Sciences International Inc. 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The Audubon Society Field Guide to North American Birds. Eastern Region. New York, NY: Alfred A. Knopf, Inc. Burton, P. 1989. Birds ofPrey ofthe World. New York, NY: W.H. Smith Publishers. Byerrum, R.U., R.E. Eckardt, L.L. Hopkins. 1974. Vanadium. Washington, D.C. National Academy of Sciences. USEPA 6922 53 * 0 0 v- 000223 Effects on Aquatic Biota: 994 Revision. ES/ER/TM-96/R1, Martin Marietta Energy Systems, Inc. Surtie, J.W. 1977. "Effects of Fluoride on Livestock." Journal ofOccupational Medicine. 19(I):40-48. Tyson, E.L. 1950. "Summer Food Habits of the Raccoon in Southwest Washington." J. Mammal., 31:448-449. U.S. Environmental Protection Agency (U.S. EPA). 1981. An exposure and risk assessmentfor arsenic. Office of Water Regulations and Standards, Criteria and Standards Division, Washington, D.C. EPA-440/4-85-005. U.S. Environmental Protection Agency (U.S. EPA). 1985. Ambient water quality criteriafor arsenic. 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"Lead in Soil." Lead inSoil Task Force, Science Reviews. Northwood. 132 pp. Woolson, E.A. 1975. Arsenical pesticides. ACS Ser7:l -176 (as cited in Eisler. 994). USEPA 6924 59 000224 USFW Calder, W.A., and E.J. Braun. 1983. "Scaling of osmotic regulation in mammals and birds." American Journal of Physiology, 244: R601-R606. Castranova, V., L. Bowman, and J.R. Wright. 1984. "Toxicity of Metallic Ions in the Lung: Effects on Alveolar Macrophages and Alveolar Type II Cells." J. Toxicol. Environ. Health 13:845-856. Clark, D.R. Jr. 1979. "Lead Concentrations: Bats vs. Terrestrial Mammals Collected near a Major Highway." Environ. Sci. Tech., 13:338-341. Dean, C.E., B.M. Hargis and P.S. Hargis. 1991. "Effects ofZinc Toxicity on Thyroid Function and Histology in Broiler Chicks." Toxicol. Letters, 57:309-318. DeGraaf, R.M. and D.D. Rudis. 1983. NewEngland Wildlife: Habitat, Natural History, and Distribution. Amherst, MA. The University of Massachusetts Press. DeMayo, A., M.C. Taylor and K.W. Taylor. 1982. "Effects of Copper on Humans, Laboratory and Farm Animals, Terrestrial Plants and Aquatic Life." CRC Critical Reviews in Environmental Control, 12(3):183-255. Dixon, R.L., R.J. Sherins, and I.P. Lee. 1979. "Assessment of Environmental Factors Affecting Male Fertility." Environmental Health Perspectives. 30:53-68. Edwards, C.A. and J.R. Lofty. 1977. The Biology ofEarthworms. John Wiley and Sons, New York, NY. Ehrlich, P.R., D.S. Dobkin, and D. Wheye. 1988. The Birders Handbook. Simon and Schuster, Fireside. New York. 785 PPEisler, R. 1986. "Chromium Hazards to Fish, Wildlife, and Invertebrates: a Synoptic Review." U.S. Fish and Wildlife Service Biological Report, 85(1.86). 60p. Eisler, R. 1988a. "Arsenic Hazards to Fish, Wildlife, and Invertebrates: A Synoptic Review." U.S. Fish and Wildlife Service Biological Report 85(1.12). Eisler, R. 1988b. "Lead Hazards to Fish, Wildlife, and Invertebrates: A Synoptic Review." UnitedStates Fish and Wildlife Biological Report. 85(1.14). Evan, A.P. and W.G. Dail. 1974. "The Effects of Sodium Chromate on the Proximal Tubules of the Rat Kidney." Lab. Invest., 30:704-715 In: Steven, J.D., L.J. Davies, E.K. Stanley, R.A. Abbott, M. Inhat, L. Bidstrup, and J.F. Jaworski. 1976. "Effects of Chromium in the Canadian Environment." Nat. Res. Counc. Can., NRCC No. 15017. 168p. Fleming, W.J. and C.A. Schuler. 1988. "Influence of the Method of Fluoride Administration on Toxicity and Fluoride Concentrations in Japanese Quail." Environmental Toxicology and Chemistr. 7(10):841-846. Fleming, W.J., C.E. Grue, C.A. Schuler, and C.M. Bunck. 1987. "Effects of Oral Doses of Fluoride on Nestling European Starlings." Arch. Env. Com. Toxicol. 16(4): 483-490. Gianutsos, G. and M.T. Murray. 1982. "Alterations in Brain Dopamine and GABA Following Inorganic or Organic Manganese Administration." Neurotoxicol. 3:75-81. Goldman, E.A. 1950. Raccoons ofNorth and Middle America. Washington, DC: U.S. Fish and Wildl. Service. 54 USEPA 6923 000225 USFW 0641 90 Mdsn Z Z O r 9