Document 5b4VeDOprbJm9eaNXGjz3YEeR

PCBs In Layinq Hens Helene C. Cecil, Joel Bitman, George F. Fries, L.W. Smith and R.J. Lillie . . Animal Science Research Division United States Deoartment of Agriculture Agricultural Research Service Beltsvilie, Maryland 20705 Presented at the 164th ACS National Meeting, NYC, NY August 1972 In the nast year there have been several Instances of contamination of ooultr'( feedstuffs by PCBs. Accidental feeding of contaminated feed to commercial chicken breeders has resulted in decreased hatchability. There have been only a few controlled studies on the effects of PCBs on reproduction in chickens. SLIDE 1 - In the experiments 1 am goinq to discuss today polychlorinated biphenyls were added to a standard breeder ration and fed ad libitum to mature White Leghorn chickens. The PCB diet was fed for 9 weeks. At the end of 9 weeks, the PCB diet was withdrawn and the birds were fed the breeder ration with no PCBs-added. Throughout the entire exoeriment feed consumption, body weight gain, mortality, egg weight, eagshell thickness, ego production, fertility and hatchability were followed. Unhatched embryos were examined for teratogenic effects. Survival and body weight gain of the hatched chicks were also followed. Residues in eggs, feces and abdominal fat were determined. These first 13 aspects of the poultry phase were completed In July of this year and I will talk about these today. Item 14 Is being carried out at the present time and will not be discussed. ' MONS 043905 2 SLIDE 2 - Our experiments consisted of 12 treatment qroups with 35 hens In each group. The Aroclors are described by 4 digit numbers. The first two digits define whether it is a bi or trlphcnyl and the last 2 digits the S chlorine. All bi and triphenyls were fed at 20 ppm. In addition Aroclors 1242, 1248 and 1254 were fed at 2 ppm since thev were the subject of the accidental contaminations. The last group of hens was fed a polybrominated biphenyl containing 75X bromine. The polybrominated biphenyl Is manufactured by Michigan Chemical Co with the product name BP-6. BP-6 Is a flame retardant to be used commercially in automobile seats. Not many chickens ride in automobiles, but BP-6 Is a potential environmental contaminant. The birds were fed ad. libitum these compounds mixed in the feed. All treatment groups ate well and feed consumption and body weight gain were the same as the controls. There were very .few deaths and no deaths could be attributed to the PCBs, PCT or polybrominated compounds. SLIDE 3 - This slide shows eggs weight, on the bottom, and eggshell thickness on the top. The left hand side of the slide is the first 9 weeks when the hens were fed PCB. The right side of the slide is the time immediately following PC8 feeding when the.hens were fed clean feed.. Each point Is the mean of 10 eggs from each group. The line represents the control. There was no effect of any of the treatments on egg weight or eggshell thickness. -' HONS O'*3906  in 3 SLIDE 4 - Although egg weight and eggshell thickness were unaffected' "' * _* by the treatments, most of the 20 nrm diets decreased egg production. The left hand portion of the slide shows egg production during PCB feeding. The right hand side shows the following two months when the hens were fed clean feed. Half of the data Is shown at the top and half on the bottom of the slide. This slide shows the effects of feeding 20 oom. 20 pom 1221 and the triphenyl 5442 had no effect on egg production. All of the other treatments lowered eqg production. PCB 1232, 1242, 1248 and the polybrominated biphenyl, PBB, reduced egq oroduction to 655. PCB 1254 and 1268 reduced egg production to a lesser extent. The results during the no PCB feeding are a little difficult to Interpret. PCB feeding started In March and the PCB feed was replaced with clean feed in May. Feeding no PCB continued until July. In late spring, with the start of warm weather, egg production normally declines. The drop In egg production In the control birds due to the Increase in ambient temperature makes the interpretation difficult. However, it aooears that in most of the groups egg oroduction is returning to the expected oroduction for the early summer season. SLIDE 5 - This slide shows the fertility of eggs from hens fed the PCBs. Tho hens were inseminated with semen from control males. Each point is the mc-an for a treatment group with the line showing tho fertility of control eggs. There was no effect of the treatments on fertility. The grey area is 85 to 955 fertility and you can see that almost all of the groups including controls were in this range. MOWS 043907 . 4 SLIDE 6 - Although PCBs had no effect on fertility of eggs, hatchability was reduced dramatically with the 20 ppm level of the middle members of the Aroclor scries. The left hand side of the slide shows hatchability during the 9 week PCB feedinq. The right hand portion of the slide shows hatchability of eggs collected during the following 6 weeks when the hens were fed clean feed. Control hatchability -- at the tot of the slide -- remained about 90S throughout the experiment. Hatchability was reduced after feeding 20 opm 1254, 1232, 1242 and 1248. The decline In hatchability started to occur after 2 weeks feeding 20 ppm. 7 weeks of feeding 1254 reduced hatchability to 70S, 7 weeks on 1232 hatchability was reduced to 40". After 1242 and 1248 feeding very few eggs hatched. Hatchability with 1242 and 1248 feeding was only 8 to 10S. When the PCB feed was replaced with feed containing no PCB, right hand portion of the slide -- hatchability began to improve. After 2 weeks of clean feed there was a surprising improvement in hatchability. Hatchability of hens fed 1232 and 1254 improved Immediately. After 6 weeks on clean feed hatchability of all grouos was better than 75S. SLIDE 7 - As we have seen from the orevious slide. 20 ppm Aroclor 1232, 1242, 1248 and 1254 reduced hatchability. Feedina these Aroelors at 2 ppm did not affect hatchability. Also 20 ppm Aroelors 1221, 1268, 5442 or the polybrominated comoound BP-6 did not affect hatchability. HONS 043908  #H 5 SLIDE 8 - We decided to investigate the dramatic effects .of the PCBs on liatchability. All unhatched eqgs were examined to determine time of embryonic death and incidence of abnomalitics. We were fortunate to have Dr. Jacqueline Verrett of the Food and Drug Administration collaborate with us on this phase of the study. Dr. Verrett is an expert on the effects of pesticides on embryonic development of the chick. She and her staff examined the unhatched eggs for us and this slide and the next series of slfdes summarize their findings. This data was only obtained recently and is not included in the preprint. The time of embryonic death is shown on this slide. Unhatched egqs from hens fed 20 ppm 1232. 1242, 1248 and 1254 of the 5th to 9th week of feeding were combined. Most of the deaths occurred at the time of hatching. Of the 840 unhatched eggs shown, 150 died on the 20th day of Incubation and 430 on the 21st day. 297 of these 430 eggs had been oipoed. This means that the chick had developed to the point where it could break the shell, but was unable to break the shell completely and emerge. . SLIDE 9 - The unhatched eggs were exaimined to determine If the embryos were abnormal. Eggs from the 5th to 9th week of PC8 feeding were examined. Approximately 450 eggs were set for each group. The X unhatched Is an expression of hatchability. In the control group only 3X of the fertile eggs did not hatch and 2X of the fertile eggs were abnormal. 1232 -- 12X of the fertile eggs were abnormal 1242 -- 27X . 1248 -- 26X . 1254 --- 51 of the 450 eggs set were abnormal. All abnormalitics were in unhatched chicks. MONS 043909 SLIDE 10 - The embryos were examined for embryonic defepts. 843 dead embryos were examined. Z86 or 342 of the 843 dead embryos were abnormal. Upon gross examination of the 286 abnormal embryos, 394 abnormalities were found. The most coitmon abnormality observed Was edema. One half of the embryos had edema. Many of-the embryos had--------- large edematous cysts on the rumo and side and many had subcutaneous edema about the neck and head area.' 122 of the embryos had unabsorbed yolk. A smaller percentage had small bodies or were midgets, had external hemorrhages, especially of the shell vessels. Skinny legs, rotated ankles and short or crossed beaks were also found. Other abnormalities that occurred to a lesser extent were cleft palate, abnormal eye formations and external viscera. SLIDE 11 - This picture Is an unhatched chick after 20 days Incubation. Abnormalities Include a large edematous cyst on the rumo and unabsorbed yolk. The large edematous cyst on the rumo had been ruptured when the picture was taken. - Unfortunately 2 other slides which showed large Intact edematous sacs on the rump and back of the neck were not ready In time for this talk. However, I do have prints with me if anyone wishes to see them at a later time. . MONS 043910 7 SLIDE 12 - Ue also determined whether the PCBs In the egg had any effect on the young that hatched. Progeny survival and growth was Studied. None of the PCB treatments affected the survival of the chicks that hatched. However, growth of the chicks was Impaired. ` ; The bars to the far left of the slide, labelled C, are the body weight gains of the chicks hatched from the eggs of hens fed the control feed. A star denotes a body weight gain that Is significantly less than I: the control. The bars next to the control are 6-9 weeks PCB. The PCBs were fed to the hens for 6 to 9 weeks, eggs collected and Incubated weekly. When the chicks hatched, they were raised for 4 weeks on clean feed and '. the body weight gain determined. The data for the 6, 7, 8 and 9 weeks of PCB feeding were combined. Chicks that were hatched from eggs of hens fed 20 ppm Aroclor. 1232, 1242* 1248 or 1254 for 6 to 9 weeks gained significantly less than the control chicks. After 9 weeks the PCB in the diet of the hens was replaced with clean feed. Progeny studies were conducted on the eggs collected during the 4 weeks immediately after PCB feeding and is shown on the right hand portion of the slide. Eggs were collected and incubated weekly and the body weight gain of the chicks then determined. One week after ' PCB 1232 was removed or 2 weeks after 1254 removal from the hens diet, the chicks that hatched grew as well as the chicks from control eggs. The denletion time necessary for Improved chick growth correlates well with the time necessary for hatchability recovery to occur. Aroclors 1242 and 1248 which had the greatest effect on body weight gain of the progeny, also had the longest residual effect on body weight gain. Hens had to be fed clean feed for 4 or more weeks,for nroneny weight gains to be comparable to controls. HOMS 043911 ( ,,! A 1 _. ' !' 1 , ; ! ' ii ^1 ; 8 SLIDE 13 - Residue analysis of eggs and feces were done weekly. Only 10v of the PCBs fed appeared In the feces. Fecal GLC patterns were the same as the feed patterns Indicating that the 10X appearing In the feces were unaltered PCBs. However, with egg and body fat samples many of the less chlorinated components were removed by metabolism. ___ ___ Thus the residues of Aroclor 1221 were greatly altered while the residues of Aroclor 1268 were essentially unaltered. ........... The analysis of the body fat and eggs of Individual birds of the PCB groups are shown here. The bar graph shows the residues after feeding PCBs for 9 weeks. In general, the level of PCB In body fat at 9 weeks. Increased with Increasing chlorination of the formulation fed. The level In egg paralled the level In body fat. The exception was 'Aroclor 1268' where the level In eggs was relatively.hiqher than expected. The excretion of 1268 in the eggs could explain the low accumula- ' '' i' tlon of 1268 In the fat. The bottom of the slide shows depletion of residues after 6 weeks on clean feed. Aroclor 1242 aonears to be removed more easily than any of the other Aroclors . The depletion of residues after feeding stops depends on metabolism,' level of egg production and changes In body composition. While depletion occurred with all compounds, a thorough analysis of all factors has not been made. . There is poor correlation of the amount of residue in the egg with the teratogenic effects. PCBs 1242 and 1248 which had the greatest adverse effects did not have higher egg residues than 1254 or 1268. In addition 1232 had a very small accumulation in the cqq - only 3 ppm - but Its effects on hatchability and embryonic development were greater than 1254's effects. MONS 043912 9 At the present time the exact component which' js responsible for the deleterious effects of specific Aroclors Is unknown. LAST SLIDE PLEASE In summary: ' A. PCBs fed to White Leghorn Laying Kens . . No effect on: . .i i i 'j Feed consumption & body weight gain Mortality j Egg weight & eggshell thickness Fertility B. 20 ppm 'Aroclor' 1232, 1242, 1248 and 12S4 Reduced egg production Reduced hatchablllty | i , ! .t ; > , i j t j I Caused teratogenic effects . . Reduced body weight gain of progeny i ' C. PCB Residues Accumulate in egg at 1/2 to 1/10 dietary level Fat residue * 10 x egg ;. . Penletlon not complete after 6 weeks feeding of clean feed. 043913 HONS  '* f ' _* EXPERIMENTAL DESIGN | MATURE WHITE LEGHORN HENS FED ad libitum Il * 1 ( i PCB IN FEED 0NO PCB I"* 9 WEEKS 'Tp 6 WEEKS J 1. FEED CONSUMPTION 2. BODY WEIGHT GAIN 3. MORTALITY ` ': 4. EGG WEIGHT y' 5. EGGSHELL THICKNESS ` " 6. EGG PRODUCTION 7. FERTILITY , 8. HATCHABILITY ! 9. TERATOGENIC EFFECTS 10. SURVIVAL OF PROGENY > 11. RESIDUES IN EGGS . 12. RESIDUES IN FECES . 13. RESIDUES IN ABDOMINAL FAT 14. DRIED MANURE RECYCLED.IN DAIRY COWS o. RESIDUES IN MILK b. RESIDUES IN BODY FAT c. DEPLETION OF RESIDUES SLioe /. I i MONS 043914 . * * . .. I '* "; i .. TREATMENTS 12 GROUPS WITH 35 H^NS PER GROUP , CONTROL 1 20 ppm 1221 PCB 21% cr /AROClORS \ | Vmonsanto/ : 20 ppm 1232 PCB 32%ci -- 2 ppm 20 ppm 1242 1242 PCB 42% cl 2 ppm 20 ppm 1248 1248 PCB 48% ci 2 ppm 20 ppm 1254 1254 PCB 54% Cl .i 20 ppm 1268 PCB 68% Cl 20 ppm 20 ppm 5442 PCT bp-6 PBB 42% Cl 75% Br /bp-6 \ Vmichigan/ 1 i ' i_ SU 0 Z. HONS 043915 i SU0 3 GRAMS mm x 10" 043917 KONS i if I! * *i suoe s~ ii MONS 043918 j tf I MONS 043919 I ii1 ; DECREASED HATCHABILITY 20ppm 1232 1242 1248 1254 . NO EFFECT ON HATCHABILITY 2 ppm 1232,1242,1248,1254 20 ppm 1221 1268 5442 (pct) pbb-6 ' i . < ii ? i t .. .. . i,, ; u i i 1 > L St-ID& 7 -1 . . ) T l MONS 043920 I . m j ii ii T! , I ` 500 . EMBRYONIC DEATH ;| 400 EGGS FROM HENS FED 20ppm PCB ' C dead embryos 300 z2> Z E3 pipped but unhatched i' 200 100 I--r^_n Ii ,. ,|l! 00 2 J___ _------. 4 6 8 10 12 14 16 18 20 INCUBATION DAY J: III $UC> S i i Ti MONS 043921 I ! '! I ! #* I EMBRYONIC DEATH & ABNORMALITIES % Unhatched % Abnormal Control t < 20 ppm 1232 J ' I 20 ppm 1242 !l 20 ppm 1248 i' 20 ppm 1254 3 39 75 . 76 18 2 12 1 27 26 5 1V fi I SUQ<=r ;i I* i; Ii ;t I i HONS 043922 I i, ABNORMALITIES IN DEAD EMBRYOS 20ppm 1232, 1242, 1248 OR 1254 DEAD EMBRYOS ABNORMAL EMBRYOS NUMBER 843 286 % 100 34 ABNORMALITIES EDEMA UNABSORBED YOLK SMALL BODY EXTERNAL HEMORRHAGE SKINNY LEGS ROTATED ANKLE SHORT or CROSSED BEAK OTHERS 394 198 49 34 30 28 12 11 32 100 50 12 9 8 7 3 3 8 SLiPe- fo MONS 043923 ' ' EGG 0 FAT 66 17 48 i 80 67 93 56 42 I 26 [ 92 17 S Lt&e /3 MONS 043926 It I| SUMMARY A. PCBs FED TO WHITE LEGHORN LAYING HENS NO EFFECT ON: FEED CONSUMPTION & BODY WEIGHT: GAIN MORTALITY ' EGG WEIGHT & EGGSHELL THICKNESS FERTILITY . B. 20ppm PCB 1232,1242,1248,1254 REDUCED EGG PRODUCTION REDUCED HATCHABILITY CAUSED TERATOGENIC EFFECTS ' REDUCED BODY WEIGHT GAIN OF PROGENY C. PCB RESIDUES * | ' ACCUMILATE IN EGG AT 2 TO 10 DIETARY LEVEL FAT RESIDUE = 10 x EGG f: DEPLETION NOT COMPLETE AFTER 6 WE^KS ! I___________________________________ SL/PE n HONS 04392V