Document Ne1e6OKOYqErdGNqxyZBJBQ4w

W U.-: M /M -L "T|V,y;"X ~^ ^ '* w,1 X 4=3 X. ;inr^r t. DEPARTMENT OF ZOOLOGY P.O, DRAWER Z STATE COLLEGE, MISSISSIPPI 39762 PHONE (601) 323-4321, EXT. 305 3 May 1967 c* Mr. 1. C. Fuhrmeister (Technical Services Superintendent Monsanto Chemical Company Anniston, Alabama 3&202 Dear Mr. Fuhrmeister: wS' .<' _ YTrfn "PV 5 */ p.y N 1-& ,-T3 **' i.l) l'PwPv-3 *v\; ^ *-,-P AJ * /v ' /?. /O In my report of April 6, 1967 I presented ACh data for distressedi fish collected at Eureka Bridege on March 26. You will e5f^ ^Htf? * bJ a ,, / fa i`rA' * recall that the specimens were quick-frozen on dry ice. The results were erratic and showed marked reductions in specific activity. On the weekend of April 8-9 we collected bluegills at State College, Mississippi and took the living fish to Anniston. At Eureka 3ri:..v;e we collected 5 bluegills and 3 additional fish belonging to the genus Lauanls. Ail of these fish (both State College and Choccolocco specimens; 'were placed on dry ice in an identical manner, returned to the lab, and tested for cholinesterase levels after one week (Table l).. The samples tested were as follows: Samples 1-8. Bluegills collected from Country Club Lake at State College, Mississippi, taken to Anniston, and frozen on dry ice. Samples 9-13* Bluegills collected from Choccolocco Creek at Eureka Bridge and frozen on dry ice. Samples l4-l6. Other fish of the genus Lepomis collected from Choccolocco Creek and frozen on dry ice. DSW 162405 STLCOPCB4040978 Vr. 1. C. I\fhr:r..eister -lay 3, 1967 Page 2 TABLE 1 ; >-7ipJ- ->c* ^">1 n--".-"1 "O4 f (^g) Optical /< M ACh. Hydrolyzed Density . 20' 60' f//* .. --1 V*-'v 1 ,-VV /'* -O' i / ->'/ 1 \! ' * 2 3 . "T j3 1 6 7 \8 i ' 49.6 56.7 26.0 34.8 5S.7 . 42.6 50.0 22.5 .090 1.17 . .120 1.52 .164 2.11 .079 .114 . 1.01 1.45 .057 .048 1.72 0.6l .063 0.60 3-51 4.56 6.33 3.03 4.35 5.16 1.83 1.80 .,y ' ,S / . 1 ,V * y ! 'tivfit/ V J 10 11 12 V13 (* n1 1 7i ,1 t '<* 4,5 ' 13 ^ / r 15 22.2 30.3 30.0 29.1 85.5 38.7 21.8 61.3 .162 .044 .053' .081 .073 .113 . .062 .072 2.06 0.56 0.68 1.04 0.94 1.48 0.79 0.93 6.18 1.68 1.98 3.12 2.82 . ~ 4.44... 2.37 2.79 Specific Activity 1.00 1.30 1.81 0,86 1.24 1-53 0.52 0.51 1.82 0.48 0.57 . O.89 0.81 1.27 0.68 0.80 Average a 109.6 91.5 DSW 162406 J' STLCOPCB4040979 Mr. I. C. Fuhrmeister Mcm 5, 1967 Page 3 None that ACh levels for "both groups of fish are low but comparable with those for frozen fish reported on April 6. Also, zhe results are erratic and contain some very low values for control fish. i`he most striking thing 'is that- the' average "value" for'Controls--. ,is much lower than, those.-we-have, found .previously-using-unfrozen-.- fish .--a If the foregoing test is valid, freezing whole fish has some adverse effect on ACh levels^ Also, af this proves to be the case, the* ^Soate of Alabama has no case. We are now planning an experiment in which the brains will be removed and frozen. It may be that freezing the entire fish is the source of the problem. Weiss recommends removing brains and freezing them in a small amount of butter. He claims that enzyme levels are unaffected by this procedure. On April 29> fish which had been caged in Choccolocco Creek since March 4- were collected and brought back to State College. On April 30, 'the cholinesterase levels of these fish were determined. Each sample represents the brain of one fish (Table 2). Samples 1-6. Fish caged in Choccolocco Creek above the Monsanto Chemical Plant (3 mi. E. Boiling Spring) ` Samples 7-12. Fish caged in Choccolocco Creek downstream from, the Monsanto Chemical Plant (Bridge on Hiway 109). ' DS\N 162407 STLCOPCB4040980 Sarnie lo. Brain hTt. (*s) IA \3 {k ^5 1 u L Vi i if 8 1 ,19 T10 1 11 12 56.8 55.0 43.2 53.1 55.0 4.10 30.6 45.6 45.3 47.2 46.4 56.7 TA3LE 2 Density K AGh Hydrolyzed 201 60t Specific Activity .235 .165 .170 .220 .145 .150 3.01 2.12 2.18 2.82 1.87 1.93 9.03 6.36 6.54 8.48 5.61 5-79 2.58 . 1.82 1.87 2.40 1.60 1.65 .205 .225 .180 .205 .230 .160 2.63 2.89 2.31 2.62 2.94 2.05 7.89 8.67 6.93 7.86 8.82 6.15 2.25 ' 2.48 1.97 2.25 2.52 1.75 Average* 198.7 > 220.3 * Procedure in this test departed from usual in that the train' mixture was allowed uo set 4 hours rather than the usual 3 The effect was a slight increase in the average activity. DSW 162408 J* STLCOPCB4040981 f C -y 3, 1967 igxs ij gX" Page 5 The following data refer to GLC analyses of samples collected March 26, 1967- .... . location Monsanto Clean-Up Plant Eureka Bridge Eureka Bridge Eureka Bridge Sample 1,000 ml water Gold fish liver and spleen (3.77 g). tfhole shad (20.82 g) Shad (whole hody minus tail). (21.93 g). ' Results No methyl parathion or parathion peaks present. Two peaks found which dud not correspond with pesticides. Many peaks present (at least 17 within 25 min. from injection). High degree of interference with pesticide standards. No methyl parathion peak present. ' DDT = .904 ppm DDD = .544 ppm DDE = .536 ppm Run on GLC. No inter ference and confirmed with thin-layer chromatography. All fishes run were showing symptoms of sickness when collected. . Many peaks (as in gold fish liver and spleen). At least 20 interfering peaks within 35 minutes from injection. The following data refer to GLC analyses of samples collected April 9, 1967. ` location Sample Results c u1 west 01 Eureka Bridge 1,000 ml water DDT = trace* DDE = trace* Lindane = trace* Plus about 5 other peaks which do not match pesticide retention times. uess unan Ppb DSW 162409 STLCOPCB4040982 n Xorr'r. of inlet vest of Eureka Bridge Under Bridge at Jacks or. Shoals S ancle 000 ml water 1,000 ml water Results 8 peaks. DDT = *3 ppb DDE = .3 ppb Lindane = trace DDE = .2 ppb DDT = .4 ppb Also 6 extraneous peaks. Pesticides (standards) analyzed for: II Lindane .-.ethyl parathion w \ Heptachlor Epoxide Heptachlor ' Parathion (6 DDT (7 DDD (3 DDE - . (9) Malathion Ue did not take DO, pH, and temperatures in April since these values have remained constant and well within safe limits. LTe hope soon to have a more definitive answer concerning the effects of freezing whole fish on enzyme activity. Sincerely yours,. D2F:wb Denzel K. Fergu'fdn^ Professor of Zoology DSW 162410 STLCOPCB4040983