«3 v." r 1' J.‘ " (J nth-n ' A 1? olcm‘ J ‘ 4“ o'..~ 'v‘ii—viflt» rrfi \ v ‘3 in.“ ff"! g; q. :4». 3.? -3??? \ :zmggzms ,3' gay; . '1 mil w 51W”? -- n ‘3 . m . 3..) ‘23 2" ~ ‘5‘ fl ‘ "mm! ‘21. EFFECTS OF BlOGEE’OCHEMiIGATL FACTORS ON THE: ‘ . Accumuwmn ' F1 7.03 ‘FALLO‘UT BY LARGE- ~ Moum BASS ( élCIRO‘PTERUS SAz'LMOleES) ,. ~ Thesis fOr fheDegree of Ph, D; MICHiGAN S TATE 'uNwERsm ”Ev-EN A isPteARELu . ‘ _ 11970 7 1 - "-O.¢Q'A'al I . U,” , LIBRARY 4' r as - Michigan State University This is to certify that the thesis entitled Efc§m {3ng Bio Fa tr on the acm agems (€131 0325§egggsflog§m§¥dégfge' presented by Steven A. Sp igarelli has been accepted towards fulfillment of the requirements for Ph.D. degree in Fisheries & Wildlife N. R. Kevern Major professor Date December 7, 1970 0-169 ABSTRACT EFFECTS OF BIOGEOCHEMICAL FACTORS ON THE ACCUMULATION OF 137cs FALLOUT BY LARGE- MOUTH BASS (MICROPTERUS SALMOIDES) BY Steven A. Spigarelli The influence of selected biogeochemical factors on 137 the accumulations of Cs fallout by largemouth bass (Micropterus salmoides) was studied in six southern Michigan lakes. The lakes were selected to provide a wide range of limnological types representing most situations in southern Michigan. Individual fish characteristics (sex and weight) and time of year were tested for their effects on the accumu- lation of 137Cs fallout by Wintergreen Lake bass. No sig- 137 nificant correlation was demonstrated between Cs activity and sex of bass. However, weight was a significant factor; 137 large bass accumulated more Cs and total Cs per unit weight than small bass. The relationship between weight and 137 Cs activity was estimated to be curvilinear, with positive SIOpe up to approximately 900 gms and negative slope above 1100 gms. Steven A. Spigarelli Mean 137Cs activities of age three bass collected monthly in Wintergreen Lake from May to October, 1969 were significantly different; the maximum activities occurred in May, the minimum in August. No correlation was demonstrated 137 between monthly precipitation and Cs activities of bass. Samples of perch and sunfish from Wintergreen Lake 137 indicated a definite relationship between Cs activity and the feeding behavior of fish; accumulation of 137Cs was greater in the more piscivorous fish species (perch > bass > 137 sunfish). A reduction in the levels of Cs from 1968 t0' 1969 was observed in all three species. Mean 137Cs activities of age three bass collected in the six study lakes during June, 1969 were compared and a multiple regression analysis was performed to determine the correlation between water parameters (Na+, K+, 137Cs, Cs+, and specific conductance) and the accumulation of 137Cs by bass. Simple correlation coefficients indicated inverse relationships between (Cs+), (Na+) and specific conductance of water and 137 Cs activity of bass, with specific con- ductance providing the best linear fit (R2 = -0.8716). Multiple regression analysis altered the relative importance of each independent variable, and resulted in the deletion of (Na+) and specific conductance from the model due to lack of significant addition to the multiple correlation co- efficient. The estimated linear prediction equation is: 137C8 = 1.5912 + 6.8118 (137Cs) - 0.0557(Cs+) - 0.3838(K+)- EFFECTS OF BIOGEOCHEMICAL FACTORS ON THE ACCUMULATION OF 137CS FALLOUT BY LARGE- MOUTH BASS (MICROPTERUS SALMOIDES) BY 1‘ (3. Steven A. Spigarelli A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Department of Fisheries and Wildlife 1970 AC KNOWLEDGMENT S I wish to express my gratitude to all persons who have been helpful in expediting my research. I wish to thank the anonymous fishermen who donated fish for analysis; the graduate students who aided my field collections; Dr. D. E. Ullrey for his help in the flame emission analyses; and Dr. J. L. Gill for his aid in the statistical analyses of the data. In addition I want to thank the members of my graduate committee: Dr. R. C. Ball, Dr. E. W. Roelofs, Dr. W. E. COOper and especially Dr. N. R. Kevern whose advice and guidance were invaluable in my graduate training. For her patience and enduring support, I owe special thanks to my wife, Betty. Lastly, I am very grateful to the U.S.A.E.C. (coo-1795-2), O.W.R.R. (A-014-MICH) and the MSU Agricultural Experiment Station for their financial support of my research under grant contracts. ii TABLE OF CONTENTS Chapter INTRODUCTION . . . . . . STUDY SITES . . . . . . EXPERIMENTAL DESIGN . . . METHODS . . . . . . . WINTERGREEN LAKE STUDIES . . Individual Variation Monthly Variation . Species Differences Annual Variation . LAKE COMPARISONS . . . . Effects of Lake Parameters SUMMARY . . . . . . . REFERENCES CITED . . . . APPENDICES A. LITERATURE REVIEW . . 137 137 Cesium Metabolism . 137Cs Uptake by Fish Cs Fallout . . B. STUDY SITES . . . . Cable Lake . . . Dewey Lake . . . Fair Lake . . . Pine Lake . . . iii Cs Flow in the Ecosystem Page 10 12 14 14 21 32 34 37 40 46 49 55 57 59 61 64 69 71 71 72 72 Chapter Wintergreen Lake . . . . . . . . 73 Schnable Lake . . . . . . . . . 73 C . METHODS AND MATERIALS . . . . . . . 7 5 Water Chemistry . . . . . . . . 75 Fish Collection . . . . . . . . 80 Fish Digestions . . . . . . . . 87 Cesium Isotope Determinations . . . 95 D 0 DATA TABLES O O O O O O O O O O O 1 0 0 iv Table 1. 10. 11. LIST OF TABLES Geographic and Precipitation Data for Six Study Lakes . . . . . . . . Limnological Characteristics of Study Lakes During 1967-1968 . . . . . Limnological Characteristics of Study Lakes During 1969 . . . . . . . Mean l37C3 Activities of Weight Groups of Bass (Wintergreen Lake, May, 1969) I O O O O O O O O O 0 Analysis of Variance for the Effects of Sex and Weight on 137Cs Accumulation by Bass (Wintergreen Lake, May, 1969) . . . . . . . . . . . 137 . . . Mean Cs Act1v1t1es and Cs Concen- trations of Age Three Bass During May-October, 1969 (Wintergreen Lake) . . . . . . . . . . . Analyifig of Covariance for Month Effect on Cs Accumulation by Bass--Weight and Precipitation as Covariates (Win- tergreen Lake, 1969) . . . . . . Stomach Contents of Bass Collected from Study Lakes During 1968 and 1969 . . Trophic Concentrations of 137Cs (Winter- green Lake, June, 1969) . . . . . Mean 137Cs Activities of Fish Species During June of 1968 and 1969 (Winter- green Lake) . . . . . . . . . Comparisons of 137Cs and Total Cs Levels in Bass and Water of Six Study Lakes (June, 1969) o o o o o o o o o Page 15 16 22 23 30 33 35 38 Table 12. 13. Anal sis of Variance for Lake Effect on 13 Cs Accumulation by Bass (June, 1969) O O O O O O O O O I 0 Linear Correlation Coefficients Between 37Cs Activities of Bass and Lake Parameters . . . . . . . . . Cesium-137 Activities and Cs Concentra- tions of Various Environmental Com- partments . . . . . . . . . . Biological Half-Times of 137Cs for Various Fish Species (Slow Component or Mean of Two Components) . . . . Digestion Procedure for KCFC Resin in H2804 O O O O O O O O O I 0 Thermal Ashing Procedure for Fish . . Nitric Acid Digestion Procedure for FiSh I I O I I O O O O O 0 Comparison of 137C5 Activities of Individual Bass Portions Digested by Thermal Ashing (T) and Nitric Acid (A) Procedures . . . . . . . . Blank Digestions and Controls for Nitric Acid Fish Digestion Procedure . . . Cesium Ion Exchange with Ammonium Molybdophosphate . . . . . . . Physical Characteristics and Cs Levels in Fish Collected in Wintergreen Lake 0 O O O O O O O I O 0 Physical Characteristics and Cs Levels in Bass Collected in Fine Lake . . . Physical Characteristics and Cs Levels in Bass Collected in Fair Lake . . . Physical Characteristics and Cs Levels in Bass Collected in Dewey Lake . . vi 39 41 58 63 85 88 89 93 94 96 100 103 104 105 Table D-S. Physical Characteristics and Cs Levels in Bass Collected in Cable Lake . . . 106 0-6. Physical Characteristics and Cs Levels in Bass Collected in Schnable Lake . . 107 vii Figure 1. LIST OF FIGURES Locations of Six Study Lakes (x) in Southern Michigan . . . . . . . Estimated 137Cs Activities and Total Cs Concentrations of Bass of Various Weights (Wintergreen Lake, May, 1969) Mean 137Cs Activities of Bass Relative to Precipitation Levels of Previous and Same Months (Wintergreen Lake, 1969) . . . . . . . . . . . 137 . . . Mean Cs Act1v1t1es of Monthly Bass Samples (Wintergreen Lake, 1969) . . Efficiency of Ion Exchange for 137Cs at Different Flow Rates by Potassium Ferrocyanide Resin (KCFC) from One Liter Samples . . . . . . . . Photographs of Field Apparatus for Cs Ion Exchange: (a) Complete Duplicate System; (b) Microswitch-Float Assembly Surrounded by Filter Chamber; (c) Box Containing 12 Volt Batteries and Water meters 0 O O I O O O O O O Styrene Ion Exchange Column for the Pre- concentration of Cs from Lake Water . Photograph of Nitric Acid Digestion Apparatus (5 1 and 3 l Flasks) . . viii Page 19 25 28 78 82 84 92 INTRODUCTION The implications of chronic low level contamination of the ecosystem by 137C5 fallout have been investigated only recently. Cesium-137 has a long physical half-life (30.5 years), emits both beta and gamma radiations, and is known to be biologically active. Potential deleterious eco-. logical effects of 137Cs contamination include specific physiological damage to the biota, trophic level increases in activity, and possible hazards to man as a consumer. The dynamics of Cs in the terrestrial environment have been studied by numerous workers. Bioaccumulations of 137Cs have been correlated to altitude (Nelson and Whicker, 1967); precipitation levels (Rickard, 1966; Eberhardt, 21': fi., 1969); climate and latitude (Pendleton, _e_t_§_1;., 1964); and specific feeding habits (Pendleton, _e_t__ai., 1964; Gustafson, 1967) . Radiocesium tends to be a potentially greater hazard to the high trophic level predators in the freshwater eco- sYstem than in the terrestrial system due to the increased availability of Cs to the biota and the more numerous predator trophic stages, allowing for more accumulation of CS The rate and equilibrium levels of uptake by the various compartments of a pond spiked with radiocesium were reported by Pendleton (1962). Cline (1967) and Rickard (1967) studied the uptake of 137Cs by aquatic macrophytes. Uptake by freshwater algae was determined by Williams (1960). Accumulations of 137Cs by fish from various lo- cations have been correlated with: precipitation levels and feeding activities (Davis, 1963); specific conductance of the medium (Preston, gt_§l,, 1967; Kolehmainen, §E_al., 1966); 137Cs activity of the medium (Preston, gt_al., 1967); and 137Cs activities of bottom sediments (Gustafson, 1969)., Trophic level increases of 137Cs activity have been «demonstrated by Kolehmainen, 3E_21. (1966), Gallegos and thicker (1968), Nelson (1967), Gustafson (1967), Pendleton (1962), and Hasanen and Miettinen (1963). Although organisms from many aquatic systems have been analyzed for 137Cs activity, only a few investigators hhave attempted to characterize the effects of limnological and biological factors on the uptake and accumulation of J“37Cs by fish. This study is an attempt to elucidate some 137 CNE the parameters that cause variability in Cs activity bGetween individuals and populations of largemouth bass. STUDY SITES Six eutrophic lakes in southern Michigan were chosen for this study to provide a wide range of limnological types (low to high alkalinity and conductivity) that were rela- ‘tively similar in locality, fallout deposition (precipi- ‘tation) and fish species composition. The selected lakes :represent a comprehensive range of the typical lakes in the Great Lakes area. Figure 1 depicts the location of the six study lakes 1J3 Michigan's lower peninsula. Geographic and rainfall data for each of the lakes are presented in Table 1. Physical aund chemical data from 1967 and 1968, and 1969 are recorded .113 Tables 2 and 3, respectively. The largemouth bass (Micropterus salmoides) was £3eelected as the test species due to its relative abundance 1111 all of the lakes and its position as a high level carni- V(are in the trophic scheme: thus, trophic level concen- 1:‘-:I'."zi.tions of 137Cs would be advantageous in analysis. Simi- ILEir fish species compositions in each lake were desirable 511: order to reduce bias due to differences in trOphic transfer of Cs to bass. Figure l.--Locations of six study lakes (x) in southern Michigan. LOWER PENINSULA Lake Huron Grand Rapids 0 . Lansing Lake 0 "ichigon X RX DONG" Kalamazoo O XX Lake ’31 Eric mm.m m¢.m .uUOIII me.o ea.m .ummmuuu em.e ve.m .mseuuu h¢.¢ hm.m MHSHIII oe.m mm.e mess--- me.~ m~.m has--- mm.v mm.~ .udauun mm.a me.o .umz--- mm He .omm 0mm .me m sea .mae oonmsmme cmmumumuces mm mm .mme owe .me mm.e~ zmam .2me emom~a< menaceom mm as .omm .mm .Ne om.m~.ma 3mm .zHa sauna mean mm as .omm 0mm owe NH.HH.~.H 3mm .zHe auumm name as am .mme .mm .H« m.m.m.e Seam .mme ammo magma as am .mme com .He m Seam .mme memo manmo meme mema GOHUMHflQHOOHm GOHHM>OHW IMMWWA OUSHHUMA mGOHflomm GOHUMOOQ huGSOU wxmn Housed :33 as. 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EXPERIMENTAL DESIGN The primary goal of this study was to determine the 137Cs activities of bass from southern Michigan lakes and to correlate the differences between the individuals of one lake (Wintergreen) and between lakes with physical, chemical, and biological parameters associated with the fish or system. Parameters tested for their effect on the accumulation of 137 137GB], [CS+], [K+], [Na+]. and spe- Cs by bass were: [ cific conductance of lake water; sex and weight of bass; and time of year (month). Effects of sex, weight, and time of year on the accumulation of 137Cs were determined using bass collected monthly from Wintergreen Lake. Analysis of variance and regression techniques were utilized to test the significance of these factors. In addition to monthly effects, the in- fluence of precipitation was of interest. Differences in 137 Cs activity of bass from the six study lakes were determined by analyzing fish collected from each lake during June, 1969. Multiple regression analyses were applied between the five water parameters and the 137Cs activity of bass. Data were adjusted for within-lake 10 11 parameters that proved significant, thereby statistically eliminating these sources of variation. An important aspect of this study concerned the alteration of existing techniques and the development of new techniques relative to Cs isotope analysis in environmental samples. The limitations imposed by the detection equipment (gamma scintillation and flame emission) dictated that Cs be preconcentrated prior to analysis. METHODS Determinations of dissolved oxygen, temperature, pH, alkalinity, and specific conductance profiles of each lake were conducted simultaneously with fish collection. [Na+] and [K+] of lake waters were analyzed by flame spectro- photometry. Cs isotopes were preconcentrated from lake water by means of rapid flow (15 L/hr) ion exchange, using cobalt ferrocyanide resins (KCFC), as described by Prout gt_al. (1965), Folsom and Sreekumaran (1970); Petrow and Levine (1967), and Boni (1966). Approximately 500 liters of lake water were preconcentrated per sample. Fish were collected by hook and line and electric shocking (115 volt, D.C.) and were kept frozen until analyzed. All bass were individually characterized as to sex, weight, length, and age. Stomach contents of each bass were identified and the relative volumes of each food item were measured. Individual fish were digested in concentrated nitric acid and the subsequent solution was treated with ammonium molybdophosphate (AMP) resin to collect the cesium isotopes (Smit, 1958; Feldman and Rains, 1964). The AMP residue was 12 13 dissolved in sodium hydroxide and extracted with sodium tetraphenylboron (TPB). Cesium-137 activities of water (KCFC resins) and fish (AMP resins) were determined by single channel gamma scintillation (0.662 Mev). Calculations of counting time intervals were made to ensure a minimal counting error (P < 0.05). Cesium-137 activities of water are expressed as pCi/L; fish as pCi/gm wet weight. Total Cs analyses on TPB extracts of fish were ac- complished by means of flame emission spectrophotometry (8521 A). The detection limit was about 0.1 ug Cs/ml. Cesium concentrations of water were estimated by considering the specific activities of fish and water to be equal (Nelson, 1967); i.e., 137 137Cs in H20 Cs in fish Cs in H20 ' Cs in fish Flame emission analyses of lake water samples were ham— pered by extremely low concentrations (ppt) of natural Cs and inadequate existing procedures for the preconcentration of water samples. Considerable effort is now being directed at this aspect of environmental Cs analysis. WI NTE RGREEN LAKE STUD I ES Cesium-137 activities and total Cs concentrations of all fish collected and analyzed from Wintergreen Lake during the study period are presented in Table D-l. Also included are data pertaining to sex, length, weight, and age of each specimen. Individual Variation The effects of individual fish characteristics (sex and weight) on the accumulation of Cs isotopes were tested by means of analysis of variance (unequal subclass numbers-- weighted squares of means) as described by Steel and Tory (1960). This analysis was performed utilizing data from May, 1969 in order to avoid a possible source of variation in uptake due to time. Cesium-137 activities of the weight groups (Table 4) were significantly different (P < 0.005) while sex and sex by weight interaction proved non- significant (P > 0.10) (Table 5). If sex were a significant factor, it most likely would have exerted an influence during May, just prior to spawning. Gonadal inspections of bass collected during May (and June determined that Wintergreen Lake fish spawned be- tween May 19 and June 20, 1969. During the period prior to 14 15 mo.HH mm.o H mm.e eo.o H ee.o m +ooe mm.e oa.m H om.m eo.o H oe.o e ems-coo om.e SH.H H ee.e mo.o H mm.o e mam-com oo.HH He.o H.mm.e no.0 H mm.o e ems-00¢ om.m me.o H efl.m mo.o H ee.o m mam-com Hm.m ee.o H m~.e eo.o H em.o m com v ANIQHVAmc\HumV A.m.m H HVAEm\ch A.m.m H HVAEm\HUQV UonHmca WMMWW .Dom .mm EsHmoU monmH HonEsz unmfloz .Amoma .amz .mxmq comnmnoucsz mmmn mo mmsoum uanoB mo moHuH>Huom mu nma Gmlel.v mqmfifi 16 mm mnmm.o H4909 mvHo.o ma mmam.o uouum om.m ma.o smoo.o m hmao.o coHuomuoucH om.m ssom.m vomo.o m Hmov.o msono usmHoB vm.v ha.o mmoo.o H mmoo.o xom mm.o m m m: up mm mousom .Ammmfl .hmz .oxmq cooumuoucsz when an coHumHsEsoom mo so uanoB can xom mo muommmo on» How oocmHnm> mo mHmhamcHuom wormH cows-I.m MHmGB 23 mm oona.o AdBOB mmoo.o Hm Hono.o mnucoz :Hnqu mo.~ samo.m mmao.o m mmmo.o mango: coo3umm mm.o m m .m.z mu mm monsom .Ammma .oxmq cooumnoucsz moHMHHm>oo mm coHumuHmHooum can uano3 uummmn an coHumHSEsoom monmH co uoommo space How mocMHHm>oo mo mwmmamcmll.h mqmfia 24 Figure 3.—-Mean 137Cs activities of bass rela- tive to precipitation levels of previous and same months (Wintergreen Lake, 1969). Mean pOi ”ks/gm net wt. 0.6 0.5 0.4 0.3 0.2 O.l 25 0.0 — same month — — previous month I J l 1 I.O 2.0 3.0 4.0 Precipitation (inchee/month) 5.0 6.0 26 137 that the monthly differences in Cs activities of bass were not the result of precipitation (fallout) changes with time. Gustafson (1969) states that 80% of the 137Cs ac- cumulated by fish from Red Lakes, Minnesota during 1965 was a result of that already deposited within the aquatic system, and that the contribution to the total activity of fish due to recent fallout tends to decrease as fallout de- 137 creases. Since stratospheric fallout of Cs has been decreasing since 1963 (Gustafson, 1969) it is reasonable to, assume that the contribution of previously deposited 137Cs to the total uptake by fish has increased while that of new fallout has decreased. Such a situation tends to explain the lack of influence of precipitation on the monthly vari- 137 ations in Cs activities of bass. Consequently, it appears that internal limnological changes, such as seasonal overturns, precipitation with CaCO3, and biological accumu- lation and sedimentation are affecting the availability of 137Cs within the system. 137 In order to compare monthly samples, Cs activi- ties of bass were adjusted for weight differences between the samples by means of the slope of the estimated regres- sion between weight and age three bass collected during May (i.e., Y = Y - bl(Xi-§). Mean adjusted values for adj. obs. each monthly sample appear in Table 6. The relationship 137 between month and Cs activity of bass is depicted in 27 Figure 4. The estimated regression equation was determined by means of least squares deletion analysis. The maximum 137Cs activities in bass during the months of 1969 that were studied occurred in May. Increased availability of 137Cs during May could have been the result of the spring overturn which probably occurred in late April. As a result of the complete mixing of the lake, a 13705 from the bottom sediments would considerable amount of be recycled throughout the system. Also, greater feeding activity of bass due to increasing water temperatures and impending spawning would most likely result in greater in- take of 137Cs. The decreasing 137 Cs activities of bass during June, July, and August may have been the result of bioaccumulation and sedimentation in organic matter due to increased primary and secondary production during these months. The feeding habits of bass during these months shed some light on the variable monthly uptake (Table 8). During May, 1969, 73% of the volume of identified food items was composed of fish while only 27% was composed of aquatic insects. But during June, the majority of food items were aquatic insect larvae (Odonata), while fish were of secon- dary importance in the diet. Assuming that successive trophic level accumulations of 137 137 Cs do occur in Wintergreen Lake, the decrease in Cs activity of bass from May to June may have been the result of dietary changes. During 28 Figure 4.—-Mean 137Cs activities of monthly bass samples (Wintergreen Lake, 1969). Mean .3703 Activity of Bass (pCi lqrn wet wt) 0.7 0.6 0.5 w 0.4 0.3 0.2 0.l 29 — observed y - — estimated 9 me: = 0.7l6l-O.l965 (T) + 0.0237(72) l l l l L l May June July Aug Sept Oct 30 .6636366663 606 6663 6636 6663 63666 60 66666660 66620666 6 6 6 6 6.66 6.63 6.66 6.66 6 66-6 63666606 6 6 6 6 6.66 6.66 6.66 6.66 6 66-6 6636 6 6 6 6 6.66 6.66 6.66 6.66 33 66-6 6 6 6 6 6 6 6.663 6.663 3 66-6 6366 6 6 6.66 6.66 6.66 6.66 6.63 6.63 6 66-6 66366 6 6 6 6 6 6 6.663 6.663 3 66-63 6 6 6 6 6 6 6.663 6.663 6 66-6 6 6 6 6 6 6 6.663 6.663 6 66-6 6 6 6 6 6 6 6.663 6.663 6 66u6 6 6 6 6 6.66 6.66 6.66 6.36 6 66-6 6 6 6 6 6.66 6.66 6.66 6.66 63 66um 6.66 6.63 6 6 6.66 6.63 6.36 6.66 63 66-6 e6666666e33 1.600 6:326 66 1.060 62.326 6 1.600 6:32, 6 6600 66 30> e GOHumuomm> 660666530 muoomcH £66m Umumacc< omuooaaou 6x63 mucoucou nocfioum Honssz mama 6.6663 666 6663 663666 66663 66666 2066 666663366 6666 66 66666666 6662066u-.6 63666 31 the remaining months of the sampling period, the only food item identified in bass was fish. It would seem that this change to a piscivorous diet should have again increased the 137Cs activities of bass. Speculation as to the reason for the decreased 137Cs activities during July, August, and September could include many aspects of limnological and physiological change during these months. However, one curious coincident that should be mentioned is the emergence of numerous aquatic insects during June and July. Obviously, the avail: ability of aquatic insects decreased after June as evidenced by their complete absence from the bass diet. Hasler and Likens (1963) have shown that aquatic midges remove "sig- nificant amounts" of radioactivity from lakes when they emerge as adults while other authors have shown insignifi- 137 cant removal by insects. Depending upon the Cs activity of aquatic insects and the magnitude of the hatches, such a mechanism of loss could account for a portion of the 137 decrease in Cs activity of bass during the late summer months. Such a loss would more likely represent a minute l37C5 within the lake. portion of the total "available" However, a reduction in immediate availability to higher trophic levels (fish) could result from the temporary loss of aquatic insects to the food chain. 137 Reasons for the increased Cs activity of bass during October can only be surmised. Decreasing water 32 temperatures and possible fall pulses of primary production could stimulate bass feeding activity and result in a 137 secondary period of growth and Cs accumulation. The apparent minimum 137 Cs activity in bass during August was tested by orthogonal contrasts and it was de- termined that the mean value during August did not differ significantly from those during July and September. Thus, the estimated regression appears to be a better approxi- mation of the actual response with time. <§pecies Differences Small samples of other fish species were also 137Cs activities of collected during May, 1969. The mean these species and largemouth bass are compared in Table 9. Although there is some overlap in the feeding habits of these species, they can be characterized as to feeding behavior, at least during particular life stages. Largemouth bass have been shown to be relatively omnivorous throughout their lives, with fish comprising a major per- centage of the diet of large bass (Ewers and Boesel, 1935). Moffett and Hunt (1943) state that yellow perch (E2323 flavescens) are generally omnivorous until attaining the length of about 5" whereupon fish becomes the major food item. Also, they observed that larger perch rely upon fish ,for 90% of their winter food. All perch included in the comparison in Table 9 were larger than 9" and can be con- sidered to have been piscivorous. Sunfish are commonly 33 oom +| 63.0 03.0 6 moono>HuoomcH AUHHnmnv cmHMcsm 666.6 66.6 H 63.3 6 6666636636. 66666 363366 006.3 63.0 H m6.0 6 msouo>Hch mmmm 2650800363 66666366 1.6.6 3 51663666 66663666 6636666 6636666 uouomm .ocoo 606m3 363852 ©6633nuocoo . .Ammm3 .ocSH .oxmq cooumnoucsz mo hma mo mGOHHMMucoocoo 03£monanu.m m3mHuom mu .onmq comnmuoucHBV hma GmmEII.OH m3m¢9 36 (1968) report a reduction in 137Cs activity of trout of 26% from 1967 to 1968; Preston, et a1. (1967) recorded a re- duction of about 75% in brown trout from 1965 to 1966. 137 Such differences in Cs activity of fish from one year to the next are evidently the result of annual reduc- tions in fallout and the rate of incorporation into the bottom sediments. The levels of 137Cs fallout have been de- creasing gradually since 1963, resulting in reduced avail- 137 ability to fish. The concentrations of Cs in the bottom sediments are no doubt increasing due to the continuing 137 buildup over time. The reduction in Cs activity of bass from 1968 to 1969 suggests that yearly changes in fallout deposition are affecting the long term equilibrium levels in 137 fish. Unfortunately, the contribution to the Cs body burden of fish due to that already deposited in the eco- system cannot be determined from the data. However, the lack of correlation between monthly precipitation and 137Cs activities of bass does suggest that changes in the avail- ability of previously deposited 137 Cs are responsible for short term, monthly variation and possibly for a portion of the yearly variation. LAKE COMPARISONS Measurements of weight, length, sex, age, 137Cs activity and total Cs concentration of all bass collected from each lake are presented in Tables D—l to D-6. Data per- taining to levels of Cs isotopes in age three bass that were used to compare the lakes appear in Table 11. 137Cs activities of bass were significantly The mean different for the study lakes (P < 0.001) (Table 12). In general, bass from soft water lakes (Fair, Cable, Dewey) 137Cs activities than those from medium to hard had higher water lakes (Fine, Wintergreen, Schnable). Such a rela- tionship was anticipated since the paucity of elemental nutrients in soft water should result in extensive accumu- lation by fish of any biologically active fallout isotopes. The mean concentrations of total Cs in bass did not differ greatly between Fair, Cable, Dewey, and Fine Lakes; but the concentrations were lower in Wintergreen and Schnable Lakes, indicating that the abundance of elements in the hard water lakes did have an effect on the uptake and accumulation of Cs. The specific activities of 137Cs (13 137 7Cs/Cs) in bass followed the same pattern as Cs suggesting that either 137 measurement is a valid indicator of Cs accumulation by fish. 37 38 6.6 66.6 H 66.66 666.6 66.6 H 66.6 66.6 H 66.6 6 66666666 6.6 66.6.H 66.66 666.6 66.6 H 66.6 66.6 H 66.6 6 66666666663 6.6 66.6 H 66.66 666.6 66.6 H 66.6 66.6 H 66.6 6 6666 6.6 66.6 H 66.66 666.66 66.6 H 66.6 66.6 H 66.6 6 66366 6.6 66.6 H 66.66 666.6 66.6 H 66.6 66.6 H 66.6 6 66666 6.6 66.6 H 66.66 666.6 66.6 + 66.6 66.6 H 66.6 6 6666 6-.. 6 c\6o 6 66-66 x6 6666 Ammmmv “MM“ ”w\wcv 60%m6m\mmmwv 66E\6omv wwwmch0MHUmv no 6666 666>6664 . o 6 o 066 66 6o m . 0666 666262 66666666 666 6.6.6 6_H6 .66666 .66666 mmxma abdum me mo 60663 cam mmmn :6 mam>ma mu Hmuou can mu wma mo mGOmHHMQEOUII.HH mamde 39 ommo.o Hm hmmm.o mmxma CHSHHZ om.~ «6m0.om mmam.o m mvmo.6 mmxmq cmm3umm m0.o m m .m.2 .m.© .m.m mousom .60m0a .mcshv mmmn an GOHuMHSEsoom 60 so pommmm 6x66 mom mUGMHum> mo mmeamc¢||.mH mqmds nma 4O 137Cs in lake waters demon- The concentration of strated little linear relationship with alkalinity and little direct influence on accumulation by bass. The maxi- mum concentration factor for 137 Cs (bass/water) occurred in Dewey Lake (soft water), the minimum in Schnable Lake (hard water), but no direct relationship between alkalinity and concentration factor was evident. The range of concentra- tion factors (l,300-l4,200 x) agrees favorably with those published by Pendleton (1962) for carnivorous fish. Such high accumulations by bass are probably the result of the long biological half-time of Cs and the high rate of uptake by bass due to their carnivorous diet. Effects of Lake Parameters In order to better define the combined effects of 137 water parameters on the accumulation of Cs by bass, a least squares deletion analysis was performed (MSU Computor 137Cs, [Cs+], specific conductance, [K+], and Lab) using [Na+] of water as fixed independent variables. Simple and multiple correlation coefficients (R2) are recorded in Table 13. Simple correlations indicated relatively strong linear 137 relationships between Cs activity of bass and [Cs+], specific conductance, and [Na+] of lake water, with specific 2 = -0.87l6). Such conductance having the best linear fit (R a correlation has been proposed by other authors (Koleh- mainen et al., 1966 and Preston, et al., 1967). Preston, 35 a1. (1967) and Hannerz (1968) suggested that [K+] was also 41 6+66 6666.6 - 6+6o6 6666.6 - 6666666 6666.6 + 6666.6 n 66666 "coHumsvm 206mmmummm omumEHumm 6666.6 666-666 6666.6 666-666 6666.6 666-666 6666.6 . 666-666 66 66666662 6666.6- 6666.6- 6666.6- 6666.6- 6666.6 66 666666 +62 .6606 .66 +6o +6 60666 666666 60666 6x6 mmHQMHHm> ucmpcmmmpsH Amy mHQmHHm> usmocmmma .mnmumEmnmm 0666 cam 6662 m0 mmHuH>Hpom momma cmm3umn musmHonmmoo coHumamuuoo HmmcHA-I.ma mqmda 42 inversely related to the 137 Cs activity of fish. However, the simple correlation coefficient for [K+] was the lowest of the five variables tested (R2 = -0.0549), implying that [K+] of water is not correlated in a linear manner to Cs uptake by bass. 137 The Cs activity of water would seem to reflect the total amount of 137Cs in the aquatic ecosystem which, in turn, should influence the accumulation by fish; but 137 the simple correlation coefficient between Cs in bass and 137Cs in water was relatively low (R2 = +0.3298). Thus, based on the simple correlations, it would seem that spe- cific conductance, [Cs+], and [Na+] of water are inversely correlated with 137C3 uptake by bass; 137 Cs activity of water is positively correlated; and specific conductance is the best linear predictor of the five variables tested. However, the very limited procedure of simple linear regression does not adequately explain the variation existing between the body burdens of bass from the study lakes. The multiple regression analysis (least squares deletion) resulted in a considerably altered order of im- portance for the independent variables (water parameters) as indicated by the multiple correlation coefficients for each set of variables in Table 13. Deletion of [Na+] and spe- cific conductance from the model did not reduce the multiple 2 137 R , while deletion of Cs, [K+], or [Cs+] reduced the co- efficient appreciably. 43 Therefore, the apparent effects of [Na+] and spe- cific conductance indicated by simple regressions were proved non-significant by the multiple technique. This altered ranking of effects is probably due to the relatively high correlations between some of the independent variables. Indeed, specific conductance is simply a measure of the . . + + + concentrations of natural ions such as Na , K , and Cs . In addition, the geochemical characteristics of drainage basins tend to provide proportional concentrations of simi- lar ions such as the alkali metals (Na, K, Cs, etc.). As a result, the "independent" variables are actually corre- lated to varying extents and their combined effects on the 137Cs accumulation by fish are considerably different than their apparent individual effects. 137 The combined effects of Cs activity, [K+], [Cs+] 137 of lake water on the Cs activity of bass appear reason- able when the chemical and metabolic similarities of these 137 ions are taken into account. The Cs activity of water 137 should reflect the availability of Cs to the biota and the levels in the aquatic ecosystem. The concentration of total Cs in lake water, which depends on the geology of the drainage basin, would, in turn, determine the relative per- 137 centage of Cs in the Cs "pool" and ultimately affect the 137Cs entering the biota. K+ is quite similar to amount of Cs+ in chemical and metabolic properties and may, under . . . + . extreme conditions, be substituted for Cs or Vice versa. 44 The estimated multiple regression equation for 137Cs activity of bass is: 137c5 = 1.5912 + 6.8118[137CS] - 0-0557ICS+1 - 0.3838[K+]. Thus, as an example, if the 137 Cs activity of bass were to ‘be estimated in an "average" southern Michigan lake (mean values of independent variables): 137CS = 1.5912 + 6.8118(o.15) - 0.0557(4-9) ‘ 0-3333(4-5) 0.61 i 0.023 pCi/gm wet weight. It seems unlikely that the variation in 137Cs activity of bass between lakes could be totally "explained" (R2 = 0.9999) by three linear variables. The very structure of the experimental design (six mean observations and five independent variables) lends itself to criticism due to a lack of sufficient error degrees of freedom. As variables were deleted from the model, error degrees of freedom re- mained small (< 3), causing some question as to the accuracy of the multiple correlation coefficients. Hoerl and Kennard (1970) warn that very high multiple correlation coefficients are frequently obtained under conditions of correlated in— dependent variables (non-orthogonal) and low error degrees of freedom. Thus, it must be emphasized that the reported multiple correlation coefficients may be overestimates of 45 the combined linear effects of [137Cs], [K+], and [Cs+] of water on the 137Cs activities of bass. The magnitude (R2) of the actual multiple corre- lation could approach 1.0 if the appropriate polynomial terms were included in the model to account for curvilinear responses; this was not attempted because of the restricted number of error degrees of freedom. If this type of study were to be repeated to better approximate the actual mag- nitude and type of correlations existing between the lake parameters and 137Cs accumulation by fish, a more extensive series of lakes should be sampled to provide a better representation of existing lake conditions and thereby in- crease the number of observations and degrees of freedom. SUMMARY The factors that affect the accumulation of 137Cs fallout by largemouth bass have been studied in a group of southern Michigan lakes. The influences of sex, weight, and time of year were tested in Wintergreen Lake; the effects of limnological parameters were determined by com- 137Cs activities of bass collected from the six paring lakes during June, 1969. The following conclusions are supported by the data: 1. A correlation between weight and Cs accumulation by bass was detected in the May, 1969 sample. The rela- tionship was estimated to be curvilinear in that an increase in body burden with increased weight (positive slope) Was characteristic of age two and three bass (200-900 gms) but the opposite (negative slope) was characteristic of bass weighing over 1100 gms. 2. No significant relationship was demonstrated between the sex and 137Cs activity of bass. 3. The 137C3 activities of age three bass decreased from May to September, 1969. A small increase in body burden was noted during October. The decrease in 137Cs 46 47 activity with time through September has not been adequately explained by diet changes. 4. A definite pattern of concentration of 137Cs between trophic levels was indicated. The most piscivorous 137Cs activi- species, yellow perch, contained the highest ties while the omnivorous bass were intermediate in activity and the insectivorous sunfish had the lowest body burdens. 5. The levels of 137 Cs in all species of fish collected from Wintergreen Lake decreased between 1968 and 1969. Such a consistent decrease in the species analyzed indicates that the input of 137Cs due to fallout is de- creasing with time and that the previously deposited 137Cs is accounting for an increasingly greater percentage of the total body burden of fish. 6. The mean 137Cs activities of age three bass collected from the six study lakes were significantly different and comprised a range of 0.10 - 1.11 pCi/gm wet weight. A general inverse relationship was observed between the alkalinity of the water and the accumulation of 137Cs by bass. 7. Simple correlation coefficients implied a strong inverse linear correlation between specific con- ductance of lake water and 137Cs uptake by bass. However, the multiple regression analysis demonstrated that this apparent linear correlation was of secondary importance 48 when all five independent variables were included in the analysis. The final prediction equation included [137Cs], [Cs+] and [K+] of water and apparently accounted for about 99% of the variation between lakes. REFERENCE S C ITED REFERENCES CITED Amphlett, C.B. and L.A. McDonald. 1956. Equilibrium studies on natural ion exchange minerals: I. Cesium and strontium. J. Inorg. Chem., 2: 403-414. Anghileri, L.J. 1960. Study of the contamination and ab- sorption of Sr-90 and Cs-l37 by Prochilodus platensis (shad). U.S.A.E.C. Report HW-tr-41. Ball, R.C. 1948. Relationship between available fish food, feeding habits of fish and total fish pro- duction in a Michigan lake. Mich. State Univ. Agric. Expt. Stat. Tech. Bull. 206. Barker, F.B. 1958. Factors affecting the transport of radioactivity by water. J. Amer. Water Works Assoc., 50: 603. Blincoe, C. 1962. Ashing procedures for determination of cesium in plant and animal tissues. Anal. Chem., 34(6): 715-716. Boni, A.L. 1966. Rapid ion exchange analysis of radio- cesium in milk, urine, sea water, and environmental samples. Anal. Chem. 38(1): 89-92. Bovard, P. and A. Gravby. 1966. The fixation of radio- nuclides from atmospheric fallout in peat bog Sphagnum sp., Polytrichium, and Myriophyllum. Radioecological Concentration Processes. Pergamon Press, London. 1040 pp. Cline, J.F. 1967. The effects of substrate conditions on the uptake rate of Cs-l37 by plants. Proc. 2nd Nat. Symp. Radioecology: 547—552. Comar, C.L. 1955. Radioisotopes in Biology_and Agricul- ture: Principles and Practices. McGraw-Hill, New York. 233 pp. Davis, J.J. 1963. Cesium and its relationship to potassium in ecology. Proc. lst Nat. Symp. Radioecology: 539-556. 49 50 Eberhardt, L.L., W.H. Rickard, C.B. Cushing, D.C. Watson and W.C. Hanson. 1969. A study of fallout cesium- 137 in the pacific northwest. J. Wildl. Mngmt. 33(1): 103-112. Ewers, L.A. and M.W. Boesel. 1935. The food of some Buckeye Lake fishes. Trans. Amer. Fish. 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Cesium-137 in freshwater fish during 1954-1965. Proc. 2nd Nat. Symp. Radio- ecology: 249-257. Gustafson, P.F. and J.E. Miller. 1969. The significance of Cs-l37 in man and his diet. Health Physics 16: 167-183. Hannerz, L. 1968. The role of feeding habits in the ac- cumulation of fallout Cs—l37 in fish. Report of Institute Freshwater Research, Drottingholm. 48: 112-119. 51 Hasanen, E., S. Kolehmainen, and J.K. Miettinen. 1967. Biological half-time of Cs-l37 in three species of fresh water fish: perch, roach, and rainbow trout. Radioecological Concentration Processes. Pergamon Press, London. 1040 pp. Hasanen, E. and J.K. Miettinen. 1963. Caesium-137 content of fresh water fish in Finland. Nature. 200(4910): 1018-1019. Hasler, A.D. and G.E. Likens. 1963. Biological and physi- cal transport of radionuclides in stratified lakes. Proc. 1st Nat. Symp. Radioecology: 463-470. Hoerl, A.F. and R.W. Kennard. 1970. Ridge regression: Applications to nonorthogonal problems. Techno- metrics 12: 69-82. Jaakkola, T. 1967. Fe-SS and stable iron in some environ- mental samples in Finland. Radioecological Concen- tration Processes. Pergamon Press, London. 1040 pp. Kevern, N.R. 1964. Strontium and calcium uptake by the green alga, Oocystis eremosphaeria. Science 145 (3639): 1445-1446. Kevern, N.R. 1966. Feeding rate of carp estimated by a .radioisotOpic method. Trans. Amer. Fish. Soc. 95(4): 363-371. Kevern, N.R. and N.A. Griffith. 1966. Effect of trophic level on radionuclide accumulation by fish. O.R.N.L.-4007: 88. Kirk, R.E. 1968. Experimental Design: Procedures for the Biological Sciences. Brooks-Cole, Belmont, Calif. 577 pp. Kolehmainen, S., E. Hasanen, and J.K. Miettinen. 1966. Cs-137 levels in fish of different limnological types of lakes in Finland during 1963. Health Physics 12: 917-922. Kometani, T.Y. 1966. Effect of temperature on volatili- zation of alkali salts during dry ashing of tetra- fluoroethylene fluorocarbon resin. Anal. Chem. 38(11): 1596-1598. 52 Krieger, H.L., B. Kohn, and S. Cummings. 1967. Deposition and uptake of Sr-90 and Cs-l37 in an established pasture. Radioecological Concentration Processes. Pergamon Press, London. 1040 pp. Krtil, J. 1965. Ion exchange of Cs and Rb on tungsten ferrocyanide. J. Inorg. Nucl. Chem. 27: 233-236. Martin, A. and R.L. Blanchard. 1969. The thermal vola- tilization of cesium-137, polonium-210 and lead-210 from in vivo labelled samples. Analyst 94: 441-446. Moffett, J.W. and B.P. Hunt. 1943. Winter feeding habits of bluegills, Lepomis macrochirus, and yellow perch, Perca flavescens, in Cedar Lake, Washtenaw Co., Mich. Trans. Amer. Fish. 800.: 73: 231-242. Myttenaere, C. and P. Bourdeau. 1967. Cesium-137, stable Cs and potassium in lowland rice. Proc. 2nd Nat. Symp. Radioecology: 553-555. Nelson, D.J. 1967. Cesium, cesium-137, and potassium con- centrations in white crappie and other Clinch River fish. Proc. 2nd Nat. Symp. Radioecology: 240-248. Nelson, W.C. and F.W. Whicker. 1967. Cesium-137 in some Colorado game fish, 1965-66. Proc. 2nd Nat. Symp. Radioecology: 258-265. Overman, R.T. and H.M. Clark. 1960. Radioisotope Tech- niques. McGraw-Hill, New York. 476 pp. Palmer, H.E. and T.M. Beasley. 1967. Fe-SS in the marine environment and in people who consume ocean fish. Radioecological Concentration Processes. Pergamon Press, London. 1040 pp. Petrow, H.G. and H. Levine. 1967. Ammonium hexacyanocobalt ferrate as an improved inorganic exchange material for determination of cesium-137. Anal. Chem. 39(3): 360-362. Pendleton, R.C. 1962. Accumulation of cesium-137 through the aquatic food web. U.S.P.H.S. Pub. 999-WP-25: 355-363. Pendleton, R.C., R.D. Lloyd, C.W. Mays, and B.W. Church. 1964. TrOphic level effect on the accumulation of cesium-137 in cougars feeding on mule deer. Nature 204(4959): 708-709. 53 Preston, A., D.F. Jefferies, and J.W.R. Dutton. 1967. The concentrations of cesium-137 and strontium-90 in the flesh of brown trout taken from rivers and lakes in the British Isles between 1961 and 1966: the variables determining the concentrations and their use in radiological assessments. Water Re- search 1: 475-496. Prout, W.E., E.R. Russell, and H.J. Groh. 1965. Ion ex- change absorbtion of cesium by potassium hexacyano- cobalt (II) ferrate (II). J. Inorg. Nucl. Chem. 27: 473-479. Rice, T.R. 1961. The role of phytoplankton in the cycling of radionuclides in the marine environment. Proc. lst Nat. Symp. Radioecology: 179-186. Rickard, W.H. 1966. Accumulation of Cs-137 in litter and understory plants of forest stands from various climatic zones of Washington. Radioecological Concentration Processes. Pergamon ifess, London. 1050 pp. Rickard, W.H. 1967. Cesium-137 in Cascade Mountain vege- tation--l966. Proc. 2nd Nat. Symp. Radioecology: 556-560. Rosenthal, G.M. Jr., D.J. Nelson, and D.A. Gardiner. 1965. Deposition of strontium and calcium in snail shell. Nature 207: 51-54. Smit, T. van R. 1958. Ammonium salts of heteropoly acids as cation exchangers. Nature 18: 1530-1531. Sreekumaran, C., K.C. Pillai, and T.R. Folsom. 1968. The concentration of lithium, potassium, rubidium, and cesium in some western American rivers and marine sediments. Geochimica et Cosmochimica Acta 32: 1229-1234. Standard Methods: for the examination of water and waste water. 1965. A.P.H.A., A.W.W.A., W.P.C.F., New York. 769 pp. Steel, R.G.D. and J.H. Torrie. 1960. Principles and Pro- cedures of Statistics. McGraw-Hill, New York. 481 pp. Townsley, S.J. 1963. The effect of environmental ions on the concentration of radiocesium by euryhaline teleosts. Proc. lst Nat. Symp. Radioecology: 193-199. 54 Wasserman, R.H. and C.L. Comar. 1961. The influence of dietary potassium on the retention of chronically ingested cesium-137 in the rat. Rad. Res. 15(1): 70-77. Whicker, F.W., W.C. Nelson, and A.F. Gallegos. 1968. Cesium-137 in trout during 1968. Annual Progress Report, UOSOAOEOCI: 24-290 Williams, L.G. 1960. Uptake of cesium-137 by cells and detritus of Euglena and Chorella. Limnol. and Ocean. 5(3): 301-311. Williams, L.G. and Q. Pickering. 1961. Direct and food chain uptake of cesium-137 and strontium-85 in bluegill fingerlings. Ecology 42: 205-206. APPENDICES APPENDIX A LITERATURE REVIEW APPENDIX A LITERATURE REVIEW The physiological and ecological implications of radioactive isotopes in the environment have only been seriously recognized within the past 25 years. As a result of the ominous destruction by atomic weapons and the con- comitant release of radionuclides to the environment, con- siderable research effort has been directed at the bio- geochemical cycling of the artificially created isotopes. However, serious gaps exist in the knowledge concerning the flow of biologically active radionuclides within the eco- system. In addition, some of the more dangerous and long lived nuclides are isotopes of relatively rare elements, such as cesium, which have not adequately been characterized, metabolically or ecologically. Cesium, discovered in 1860, is the most alkaline and chemically active of all metals. The primary mineral source of cesium is pollucite, a hydrated silicate of aluminum and cesium, containing approximately 30% C50. Cesium exists as a trace element in most forms of rock (1- 4 ppm), coal (~ 13 ppm) and soil (0.1-25 ppm). The concen- tration of cesium in sea water is relatively constant 55 56 throughout the earth (0.3 - 1.0 ppb) while that in fresh water may vary considerably depending upon the geology of the drainage area (1 - 100 ppt). Twenty-one isotopes of cesium are known (123Cs to 144 133 Cs), the stable isotope being Cs (atomic weight = 132.905, atomic number = 55). All other isotopes of cesium are radioactive and are artifically produced as a result of a variety of nuclear reactions. The biological importance of these many isotopes depends upon their: (1) metabolic activity, (2) physical half-life, (3) relative abundance as radioactive pollution, and (4) emission forms and energies. Cesium-137 is probably the most important of all fallout isotopes, considering all aspects of its activity. It has a very high fission yield (6 atoms/100 fissions) and occurs 90 at a rate of 1.5 times that of Sr (Gustafson and Miller, 1969). Cesium-137 is formed as a result of the following decay scheme: 137I 19E 137Xe§+4_, 137,35 104.5, 137-as (stable). min. min. yrs. 137 The 30.5 year physical half-life of Cs results in a high potential for biological accumulation. An energetic gamma emission (0.662 mev) and two beta emissions (0.514, 1.18 mev) present considerable hazard to organisms. The bio- 137 logical and metabolic similarity of Cs to stable cesium and potassium imply considerable activity within the eco- 133 system. Cesium-l37 activities and Cs concentrations of 57 various environmental compartments are listed in Table A-1. Cesium-134, a minor component of fallout, could become an abundant radioactive pollutant as a result of nuclear re- actor irradiation of stable cesium or 136Ba. 137Cs Fallout Although the major source of 137 Cs pollution is stratospheric fallout resulting from thermonuclear deton- ations, other potential sources that exist are: (1) loss from the chemical separation of reactor fuels, (2) acci- dental high-level discharge from reactors (effluents or aerial), and (3) low-level discharge from reactors. To 137Cs have been minimal date, these secondary sources of despite detection of traces of the isotope in reactor effluents and infrequent accidental releases. However, the trend toward nuclear power reactors suggests that concern over the potential hazards to the environment is not un- warranted. Stratospheric fallout of 137Cs, as well as other radioisotopes began as a result of the advent of thermo- nuclear weapons in 1952. Variations in global fallout of 137C8 depend primarily upon latitude, the maximum levels occurring at 45° N, the minimum at the equator (Davis, 1963). Altitude can also be a determining factor, fallout levels increasing with elevation (Nelson and Whicker, 1967). Seasonal patterns of high and low fallout are common, the highest levels occurring in the spring and frequently in 58 6666 .606666666 66666 666 6666 66 - 6.6 6666 ..H6 66 .66666666666 6662 6666 6666 6.6 66666666 6666 6666 ..H6 66 .66666666606 6666666 6666 6.66 - 6.6 60666 6666 ..66 66 .66666666606 6666666 6666 6.66 - 6.6 6666 .606666666 66666 666 6666 6.6 - 6.6 6666 6666 .606662 .6 606666 6666 66.6 6-66 6 6.66 6666666 6666 .6666666 666636 6666 6.6 - 6.6 6666 66663 6666 ..H6 66 .6066666 6666666 6666 6.6 - 66.6 66066 63066 6666 ..66 66 .66666666606 6666666 6666 6.6 - 6;6 6666 .606666666 6660666606 6666 6.6 - 6.6 66066 6666 6666 .606666666 66666 666 6666 6.6 6666 .3.6 6666 ..66 66 .66666666666 6662 6666 6666 6-66 x 6.6 6666 .606662 .6 606666 6666 6-66 x 6.6 6-66 x 6.6 6666 ..66 66 .66666666606 6666666 66-6666 6-66 x 6.6-6.6 6666 .606666660 66666 666 6666 6-66 6 6.6V 66663 66666 6666 .606666666 6666 6.6-66.6 6666 .606606 0666066 66-6666 66.6 6-66 x 66-6 6666 6666 .606606 0666066 66-6666 6-66 x 6- 60666666 6666 .606606 0666066 66-6666 6-66 x 6.6 66666 63066 6666 .606606 0666066 66-6666 6-66 6 66.6 6-66 x 6.6 66663 666 6666 .66>66 6.6- 6006 .666 6666 .66>66 6.6- 6666666 6666 .66366 66- 6606 6666 .66366 66 - 6.6 6606 mocmnmmmm co6umooq mpmo AEm\HUmv Aammv mu ucmEuummEoo . 606ma 666c666066>cm .mgcmauumm 1500 Hapsmesoufi>cm 650666> mo mGOHUMHucmocoo mo was mmwufl>6uom find-Edwmmoll.alm unmfla 59 the fall. However, seasonal patterns and altitudinal vari- ations in fallout are most likely reflections of precipi- tation levels. There are numerous reports of positive correlations of 137 Cs fallout (and biological accumulation) with precipitation levels (Rickard, 1966; Eberhardt, gt_§l., 1969). Davis (1963) states that stratospheric 137Cs is mostly dissolved in water droplets (70% is water soluble) which supports the relationship between precipitation levels 137 137 and Cs fallout. Maximum fallout of Cs occurred during 1963 (Gustafson, 1969). 137Cs Flow in the Ecosystem Cesium-137 fallout deposited in the terrestrial environment tends to be primarily adsorbed to upper stratum soil humus particles and clays with very little transfer to plants (Rickard, 1966). The primary route of entry to the terrestrial food chain is through direct deposition on foilage, which in turn serves as food for other organisms. 137 Consequently, a large proportion of the incident Cs is not available to the organisms of the system. Some ex- ceptional situations do exist, however, in the cases of bog areas and tundra where vegetation characteristically con- centrates many elements. Pendleton, gt_al. (1964) suggests 137 that the relatively high Cs body burdens of Alaskan Eskimos and Laplanders are due to a combination of factors: (1) the climatological conditions, latitude, precipitation, 137 etc. favor rapid accumulation of Cs by lichens; (2) 60 cesium is concentrated by a factor of 3 x from one trophic level to the next (lichen-caribou-man); and (3) the people of these regions tend to be strictly carnivorous, obtaining their food from trophic levels in which cesium was greatly concentrated. Major sources of 137 Cs to other human popu- lations are meat, milk, grain, and freshwater fish. In the aquatic environment radiocesium is poten- tially a greater hazard for two reasons: (1) the water medium increases the availability of cesium to the biota, and (2) the more complicated food chains involve many more trophic levels than in the terrestrial system, allowing for more concentration steps. Cesium-137 deposited by fallout is rapidly removed from water by sediment, colloidal clays, and the biota. The partitioning of cesium between sediment and water may be pH dependent (Gallegos and Whicker, 1968) or the exchange capacity of clays may be constant above pH 3.5 (Amphlett and McDonald, 1956). Barker (1958) states that the percent Cs adsorbed by clays decreases as concen- trations of K+ and Na+ increase, which implies that more 137Cs would be available to the biota under hard water con- 137 ditions. Adsorption of Cs to soils in bogs is depressed by the acid medium, resulting in increased availability to bog plants (Bovard and Gravby, 1966). Uptake of 137 Cs by aquatic plants has been shown to be considerably greater than by terrestrial plants (Pendle- ton, 1962; Cline, 1967; Rickard, 1967). Uptake by algae is 61 137 almost immediate and linear with Cs concentration of the water (Williams, 1960). Also, marine biota accumulate considerably less 137Cs than do freshwater organisms. Rice (1961) has shown that marine algae do not accumulate 137Cs to any degree probably due to the high concentration of stable Cs and other ions in sea water. Also, it has been stated by Williams (1960) that increasing concentrations of Cs inhibit the uptake of 137Cs by algae. Cline (1967) de— fines the conditions which exist in the aquatic environment that favor the uptake of cesium by plants: (1) Cs is free in the medium as ionic form, (2) absorption surfaces of plants are in liquid phase with Cs, and (3) there is turbulent mass flow within the solution. Thus, given equal contamination, the availability 137 and accumulation of Cs tends to be greater in the fresh- water ecosystem than in the terrestrial or marine systems. Cesium Metabolism Although Cs has long been equated with K metaboli- cally, recent evidence suggests that, aside from basic simi- larities, these two elements differ in their metabolic roles. The similarities between them have resulted in wide- spread use of Cs/K ratios by radioecologists, whereby, the accumulations of cesium are frequently "explained" by the relationships between Cs and K. However, sufficient evi- dence exists that stresses the differences between Cs and K uptake and metabolism to warrant the treatment of Cs as a 62 discrete metabolite. Williams (1960) states that algae cells do discriminate between Cs and K and that the [K+] has little effect on Cs accumulations. Davis (1963) observes that marine algae may fulfill a K+ deficiency with Cs but they in turn are able to discriminate between them. Wasser- man and Comar (1961) state that changes in dietary K+ have little effect on Cs retention by organisms, but Cs and K+ do most likely share the same transport mechanisms. Perhaps the most striking difference between Cs and K metabolism is their rate of excretion or biological re- tention times. Most organisms preferentially retain Cs within cells much longer than K, resulting in greater effective concentrations of cesium over the ionic source. Biological half-times (TBl/2) of Cs for some common fish are listed in Table A-2. The primary route of uptake of Cs by freshwater fish is through food, but some marine molluscs and fish and freshwater crustaceans (Daphnia) can accumulate Cs directly from the water. Absorption of Cs from the gut is rapid and complete. However, the overall metabolic flow of Cs is con- siderably slower than that of K. The excretion of Cs in fish follows two exponential phases, a fast fraction (1-- few days) and a slow fraction (few--500 days) (Hasanen, g£_31., 1967). In the same study, old fish were shown to exhibit longer TB l/2's than young fish of the same species. 63 6666 ..66 66 6666 bmmd ..Hm um 6666 .mcfinmonm 0cm 6mma mama ..Hm um 6666 .6660663 666 6666 .6660663 666 mmma 66660633 026 6666 ..66 66 6806606 .snm>mx .cmsmmmm 68666663 .606662 .6066666 mommaamw 60066660 60066660 .cmcmmmm ooa v qua I mm com I mud ov 2 0mm r com 2 mm 2 mod I OOH I om I mm 6659 6660 60666 66666666 6666660 s3oum xooum 660666666 303:666 3oncwmmIIusoue mocmummmm 666666 6\6 66 mmwommm usmcomEoo 3066V 6660666 6666 650666> 60m 60 .Amusmcomsoo 036 Mo 6666 Ho bmH 60 66666-6666 6606606066-.6-6 66666 64 137 The relationship between K+ and Cs uptake by fish has been discussed by many authors usually in light of the metabolic similarities. Inverse relationships between 137Cs uptake in fish and [K+] of water have been reported by Preston, g£_al. (1967) and Hannerz (1968) but Hannerz quali- fies this relationship by stating that K is likely to be responsible for only a small portion of the variation in 137Cs uptake. Inconsistent relationships between 137Cs up- take and [K+] of water were described by Kolehmainen, 3543;. (1966) and Whicker, g£_al, (1968). Thus, it appears that a. lack of agreement exists concerning the effects of [K+] on the uptake of radiocesium by fish, but most workers agree that Cs and K are discrete metabolites. The assumption of identical chemical and biological behavior of 133Cs and 137Cs is based on the similar atomic masses of the two isotopes and experimental evidence that 137Cs/Cs) of water to be has shown the specific activity ( approximately equal to that of fish (Nelson, 1967). The importance of the stable isotope concentration of water on the uptake of 137 Cs by fish has been implied in the litera- ture (Krieger, et al., 1967; Palmer and Beasly, 1967; Myttenaere and Bourdeau, 1967; Folsom, et al., 1967). 137Cs Uptake by Fish Cesium-137 activities of fish have been decreasing recently from maximum levels observed in 1963 (Preston, 33 al., 1967; Gustafson, 1969). The corresponding activities 65 in trout from 1967 to 1968 amounted to a decrease of about 26% (Whicker, et al., 1968). Seasonal patterns of 137Cs uptake have also been postulated. Davis (1967) states that seasonal patterns of uptake are probably reflections of precipitation levels or periods of active feeding by fish. Nelson (1967) not only described temporal fluctuations of 137Cs but also of 133Cs. Cesium-137 activities in various fish are included in Table A-1. 137 The sites of deposition of Cs in fish (shad) tissues were described by Anghileri (1960). Primary sites were heart, muscle and skin (48 hrs.) while, over a longer 137 period of exposure (2 - 30 days), Cs accumulated in hardparts such as head, gills, and skeleton. The implica- tions of such ubiquitous deposition of Cs throughout the body of an organism reveal the potential hazard arising from 137Cs contamination of the environment. In addition to the effects of [K+], many physical, chemical, and biological factors have been linked with the variable uptakes and accumulations of 137 137 Cs by fish. In- verse relationships between Cs activities of fish and the specific conductance of water have been shown by Preston, EE_2l- (1967) and Kolehmainen, e£_al, (1966); no consistent correlation was detected by Nelson and Whicker (1967). Preston, gt_al. (1967) suggest a direct relation- 137 ship between the Cs activity of the water and that of the fish while Kolehmainen, et a1. (1966) were unable to show 66 such a relationship. No consistent correlation between 137 lake elevation, area, temperature, watershed area, Cs in 137 bottom sediments and Cs activities in trout could be detected by Whicker, et al. (1968). Gustafson (1969) how- 137 ever, attributes much of the variability in Cs activity of fish from Red Lakes, Minn. to the activities of localized bottom sediments. Although no relationship could be es- 137 tablished between fish size and Cs activity by Whicker, et a1. (1968), Preston et al. (1967) state that the TBl/Z tends to increase with body weight, implying that equi- 137 librium activities of Cs would also be higher. Although no correlations have been demonstrated . . 137 between ceSium concentrations of water and Cs accumu- lation by fish, such relationships have been shown for other biologically active isotopes. Palmer and Beasley (1967) 55 and Jaakkola (1966) state that the uptake of Fe by fish is inversely related to the concentration of stable Fe in the water (marine and freshwater, respectively). An inverse 89 relationship between stable Sr in water and Sr uptake by fish was suggested by Townsley (1963). However, Kevern (1964) demonstrated that increased Sr concentrations re- 85 sulted in greater uptake of Sr in green algae. A positive correlation between Sr concentration in the medium and 85Sr uptake by clams was also reported by Rosenthal, et al. (1965) . 67 137 Trophic level increases of Cs activity have been demonstrated by numerous workers in both terrestrial and aquatic systems. Kolehmainen, gt_al. (1966) found 137Cs activities of bottom feeding and predator fish to be about 2x and 4x respectively that of plankton feeders. Gallegos and Whicker (1968) have shown that trout concentrate 137Cs about 2x over their main food items (amphipods). Nelson (1967) noted that piscivorous fish contained the highest 137Cs activities although no exact relationship between trophic position and 137Cs content was evident. Gustafson (1966) calculated trophic level concentration factors of 137Cs for fish to be: perch/small fish = 1.85; pike/perch = 4.8; pike/small fish = 8.9. Pendleton (1962) and Hasanen and Miettinen (1963) noted consistently higher concentra- tions of 137 Cs in carnivorous (predator) species than in herbivorous or omnivorous species. Few differences be- tween 137Cs activities of carp and food items such as algae, or between trophic levels were detected by Kevern (1966) and Kevern and Griffith (1966). However, as indicated by other authors, piscivorous fish tend to concentrate 137Cs to a greater extent than do omnivorous species, and such a trend was shown in the study by Kevern and Griffith. The overwhelming evidence in favor of trophic level concentrations of 137 Cs present some serious ecological implications relative to human welfare. Cultures that rely heavily upon freshwater fish as food, such as Scandanavians, 68 risk high body burdens of 137Cs, especially if upper level predator fish are consumed. The advancing role of aqua- culture as a solution to food shortages may result in in- creased accumulations of radiocesium by man. Gustafson (1966) warns of the possible hazards resulting from 137Cs activities in fresh water equal to 1/10 the maximum per- 7 Ci/L). When missible concentration set by law (2 x 10- eaten in the limited quantities characteristic of our culture (8 kg/yr.), fish from such contaminated water would produce 137Cs body burdens in man (175 uCi) that are 60 times the established maximum permissible limit for the general pOpulation (3 uCi). APPENDIX B STUDY SITES APPENDIX B STUDY SITES In view of the defined goals of this study and the existing research results concerning 137 Cs uptake in fish, a series of lakes were chosen that provided: (1) a wide range of conductivities and limnological types, (2) fallout. patterns that were nearly equal due to the proximity of the lakes, (3) similar fish species and abundance, especially largemouth bass, and (4) ease of sampling. Many areas in southern Michigan were surveyed for such a series of lakes and numerous methods of gathering information were utilized: such as screening the files of the Michigan Department of Natural Resources and interviewing residents of prospective study lakes. The final selection of six study lakes was made during 1967. A complete geographic location of each lake is presented in Table 1. An inverse relationship between 137 Cs uptake in fish and the specific conductance and/or the [K+] of the water has been proposed by numerous workers: Kolehmainen, et_al. (1966), Preston, gt_§l. (1967), and Hannerz (1968). Other authors have refuted such correlations (Nelson and Whicker, 69 70 et al., 1968). The selection of study sites was designed so that these ecological parameters could be tested for 137Cs fallout by bass in their influence on the uptake of Michigan lakes. Fallout patterns for the study sites were assumed to be directly dependent upon precipitation levels and the proximity of the lakes. Davis (1961) states that the maximum levels of radioactive fallout occur at 45° N Lati— 137Cs fallout tude, the minimum at the equator, and that depends almost entirely on precipitation levels. Nelson and Whicker (1967) include latitude, elevation, precipi- tation, and the drainage area among the factors affecting the amount of fallout entering an aquatic ecosystem. Due to similar geographic locations and nearly equal annual pre- cipitation levels of the lakes (Table 1), it was expected that the input of 137 Cs fallout would be relatively uniform throughout all six lakes. The largemouth bass was selected as the test species because of its relative abundance in southern Michigan lakes and its position as an upper-level carnivore in the trophic chain (thus, trophic level concentrations of 137Cs would be advantageous in analysis). Similar species compositions of fish among all lakes were sought to reduce bias due to var- iations in feeding habits of bass or differences in trophic concentrations. 71 The accessability of each lake for onsight limno- logical surveys and the collection of bass samples was an important factor. In the case of each study lake, either the presence of a public access site or the cooperation of residents was imperative for extensive data and sample collection. Cable Lake Cable Lake (Cass County), the least alkaline of the six lakes (~ 6 ppm CaCO3), is relatively nutrient poor and slightly acidic (pH ~ 6.5). It's drainage area is composed of rural and agricultural lands, while the shoreline is almost entirely occupied by permanent dwellings. The lake has no inlet or outlets and is essentially a closed eco- system. There is no public access to Cable Lake and recre- ational usage is restricted to residents. A very influen- tial home owners association has established a stringent set of rules concerning the use of the lake, including the banning of all outboard motors. As a result, the collection of bass by electric shocking was impossible and the only recourse was to rely on the generosity of resident fisher- men. Fortunately, ample supplies of bass were provided. Dewey Lake Dewey Lake (Cass County) is somewhat more alkaline (~ 18 ppm CaC03) than Cable Lake and quite similar in nutrient composition and acidity (pH ~ 6.7). The drainage area is predominately urban, as there are considerable 72 concentrations of homes and small communities surrounding the lake. A public access site is present on the lake and a great deal of fishing and boating activity is common. Bass collection was eventually accomplished by means of electric shocking at night, since daylight attempts at shocking and hook and line fishing proved futile. Fair Lake Fair Lake (Barry County) is also quite low in alkalinity (~ 41 ppm CaCO3) but unlike Cable and Dewey Lakes, it is slightly basic (pH ~ 7.5). The draining basin is composed of a mixture of rural and wooded areas, with a considerable number of shore dwellings. No public access site is available and usage is restricted to land owners. The fish sampling of Fair Lake was accomplished as a result of the cooperation of the residents. Some of the bass samples were collected by electric shocking at night while others were donated by resident fishermen. Fine Lake -Fine Lake (Barry County) is somewhat intermediate in alkalinity (~ 85 ppm CaCO3) while being the most basic of the lakes (pH ~ 7.8). The drainage area of Fine Lake is mostly rural with a considerable number of shore dwellings. A public access site is available and the lake provides much fishing and boating recreation for the area. Bass samples were obtained with little difficulty by means of electric shocking during the day. 73 Wintergreen Lake Wintergreen Lake (Kalamazoo County) has for many years been managed as a waterfowl refuge by the Gull Lake Biological Station (MSU). The lake is quite eutrophic and its alkalinity (~ 130 ppm) is typical of southern Michigan lakes. Although the alkalinity and pH (~ 7.5) are inter- mediate, considerable amounts of inorganic nutrients are present, such as K+ (~ 13 ppm) and Na+ (~ 8 ppm). These elements are no doubt being added to the system continuously by the large numbers of waterfowl that regularly occupy the lake. Public access is completely restricted with the ex- ception of sightseeing visitors. The electric shocker was not used to collect bass, in deference to the waterfowl. Instead, hook and line methods were employed and there was little difficulty in obtaining adequate samples. Schnable Lake Schnable Lake (Allegan County) is the most alkaline of the lakes studied (~ 200 ppm CaCO3) and is of moderate pH (~ 7.7). The drainage basin consists predominately of rural and agricultural land and there are very few shore dwellings. There is no public access to Schnable Lake and its recreational use is limited to the few residents. The collection of bass samples was accomplished with some diffi- culty through a combination of day and night shocking and hook and line fishing. 74 The limnological characterizations of each lake for the three year study period are presented in Tables 2 and 3. Although the lakes vary considerably in alkalinity, specific conductance and dissolved nutrients, all six must be termed eutrophic as evidenced by their clinograde oxygen profiles. Each is typically dimictic, experiencing both spring and fall circulation periods. The shallow basins and the re- duced hypolimnitic volumes relative to the epilimnitic volume of each lake are no doubt responsible for their eutrophic condition. With the possible exception of Schnable Lake, each lake has also been receiving some do- mestic as well as agricultural drainage, adding to its natural tendency toward enrichment. Consequently, it must be noted that each of the six study sites has in no way es- caped the damaging influences of man (and waterfowl, in the case of Wintergreen Lake). APPENDIX C METHODS AND MATERIALS APPENDIX C METHODS AND MATERIALS Water Chemistry The following onsight analyses and measurements were performed at each lake at the time of fish collection: (1) depth (sounding line); (2) temperature and specific conductance (battery operated conductivity meter); (3) dissolved oxygen (Galvanic oxygen analyzer); (4) pH (port— able pH meter); and (5) alkalinity (H2804 titration, Stand- ard Methods, 1965). Water samples were collected prior to returning to the laboratory and refrigerated until analyzed. Determin- ations of [K+] and [Na+] were conducted by the water chem— istry laboratory at MSU by means of flame spectrophotometry. 137Cs and total Cs were Analyses of lake water for complicated by the extremely low concentrations of each (< 0.3 pCi/L and < 20 ppt, resp.). Initially, a procedure of cesium analysis from water samples as described by Feldman and Rains (1964) was attempted with little success. This procedure appears to be better applicable to waters containing high concentrations of Cs and 137Cs. 75 76 The sensitivity limits of the available gamma scintillation detector (~ 100 dpm/sample) and flame emission spectrophotometer (~ 0.2 ppm Cs) dictated that a consider- able amount of preconcentration of Cs from lake water was necessary for analysis. A suitable procedure for rapid flow ion exchange of Cs from lake water was deemed the most feasible solution. Numerous types of ion exchange resins have been applied to the collection of Cs from various solutions. However, most of the resins have proven to be either non- specific for Cs or to have characteristics not amenable to rapid flow column use. Various forms of complex ferro- cyanide compounds (Co, Zn, Cu, W, etc.) have been applied as ion exchange resins for alkali metals. The order of sorption of the alkali metals to these resins is Li 06 6666066>m .666666066 66 mozm 6608 666 «60665606 66 66 6666 666 66665 656666 .em\mozm .ocoo 6s m 666 .E6\6E 6 65066 06 6666066>m .66506 6 66666 66 6666606 66 6x56666v 666666 .6666660 6666606 6608 66665v 66506 cum 65066 666666 06 30664 .am\mozm .oaou 66 m 666 MQO'UCD‘HU‘S "66666 66 66066066 6666 60666 .m .666663 663 666660 .N .66666 6666606 666 06 660666 0666 6666 650 .6 .6666 606 665660066 606666666 6606 06666zun.m10 mqmda 90 in large glass boiling flasks fitted with reflux condensers 137Cs and Cs utilized (Figure C-4). All determinations of in the final statistical analyses resulted from fish that were acid digested. This procedure has: (1) substantially reduced the ashing time and the number of steps in the 137C3 and Cs determinations in analysis, and (2) yielded fish that are considerably more accurate than those obtained by dry ashing. A comparison of the two tissue ashing methods is presented in Table C-4. In this comparison, individual fish were cut into two portions and each was ashed by a different procedure. Once the two portions were put into solution, 137 each was analyzed for Cs by the same technique. The consistently lower values for 137 Cs recovery in the por- tions undergoing dry ashing support the choice of acid di- gestion in this study. Also, in order to establish the efficiency of the acid digestion procedure, a series of 137 Cs tagged solutions were "digested" and processed under the same conditions as fish solutions. Thereafter, possible losses of Cs occurring through adsorption or volatilization were determined. The results of these tests are given in Table C-5. The small apparent losses of 137Cs may well have been a result of counting errors, rather than actual losses of Cs. The ion exchange of Cs isotopes from fish ash solu- tions was performed by a batch procedure, using ammonium 91 Figure C-4.--Photograph of nitric acid diges- tion apparatus (5 l and 3 1 flasks). 92 93 m.66 mm.o 6.66 M6v 6 em.o 6.66 ~66 a m 6.66 om.o 6.66 omv 6 em.o m.om om6 a v m.wv 6m.o m.m~ 666 6 mm.o 6.66 m66 a m 6.06 6m.o 6.mm 66m 6 mv.o 6.66 mm6 a N 6.6m mo.o 6.66 mmm 6 om.o «.66 N66 6 6 66\au6v .63 663 660 Amsmv 666m .6666 m 666\606V 66609 .62 66: 606666 6666 >6 66666666 66066606 6666 66566>6666 mo 66666>6606 60 .6665660066 64v 6606 066666 666 68v 666666 6686666 hM6 mo 6066666Eoolu.vnu M6669 94 c I: c cccm c cccm m c I: c ccom c cccm m c I: c comm c ccom v I: c.cc6 hm comm mm cccm m nu v.mm m6 comm we cccm m In 6.mm cmm comm mmm cccm 6 man m66>0066 Asmuv AmEo Azmov 668v 60 w ©666>006m mohMH 624 @6664 monMH mozm 666B .666660066 606666m6o 666m 6606 066666 606 66066600 6:6 6606666660 6666muu.mu0 66666 95 molybdophosphate (AMP), a highly selective resin for alkali metals (Smit, 1958; Feldman and Rains, 1964) (Table C-6). The relative ease of collecting the AMP by centrifugation and dissolution of AMP in NaOH solution provided a simple means of concentrating trace amounts of Cs from fish into small volumes. Successive cleanup steps eliminated the high levels of K from the sample, thereby reducing contamination by 40K and facilitating the liquid-liquid extraction of Cs into organic media. AMP used in these analyses was prepared in the laboratory in order to eliminate the "background" Cs inherent in commercial AMP (Folsom and Sreekumaran, 1970). Polyethylene or polypropylene labware was used in all analytical procedures in an attempt to avoid the ad- sorption of Cs to glass. The final aqueous solution containing Cs was ex— tracted with 0.05 N sodium tetraphenylboron solution (TPB) (3/1 hexonezcyclohexane) (Feldman and Rains, 1964; Folsom, 1970). The organic layer was retained for flame emission analysis for total Cs (8521;). Cesium Isotope Determinations Cesium-137 activities of fish and water samples were determined by means of single channel gamma scintillation spectrometry (0.662 Mev). A 3" NaI crystal detector with a 1 l/4" x 2" well (Harshaw) was coupled with a spectrometer and a decade scaler. Counting vials were all of the same diameter (1") to ensure consistent counting geometry. The 96 .66666066666 68 m 036v 60 606 66666666 60666686 66666 6663 6660066 .m6 6 660 . ohm6 6 U 66 .66666 0666660 666666 “66506 6. 6666660 06 30666 .M6 .666668 6 666606066> 66666 «66962 z 6.c 65 c6 6663 60666xm .m6 .666 06 660666 6062 2 0.6 66 626 6>60666o .66 606M6 66500 “66666666666 6660666 66 6666 666666 .c6 .6666666660 «6663 6666 66666 .63 663 E6\mz¢ 6E 6.c 66¢ .m .6606 06666666 6663 m.m 06 :6 666666 .6 .momz Z 0.6 66 m2< 6>60666Q .6 60>M6 66600 65666 «626 6066600 .6 .66666666666 6660666 66666666>0 666666 06 30666 60 6666666660 .m .6666666 6 66066 6663 6666 6.63 663 66\666 .66 6 666 .6 .66666666866 8006 06 6000 06 3066< .m m .0 6 6663 6663 66666 6666606 66666 .N .666666 666666666606 66666 06 60666606 666 66666669 .6 .6666660660666606 85660856 6663 666660x6 606 55666oll.mlu mqmfia 97 efficiency of the detection system and the 137Cs energy peak were checked daily by means of a 137C3 standard. Counting efficiencies averaged 13.5% (cpm/dpm). The possible contributions of gamma rays to the 137Cs channel by 40K and 134 Cs were anticipated. However, it was determined that the narrow channel that was counted (.005 Mev) received negligible interfering radiations from other radioisotopes. Representative fish and water samples were analyzed by the Argonne National Laboratory and the Oak Ridge National Laboratory: the only radioisotopes re- ported by these labs were 137Cs, 134Cs, and 40K. Activities of 137Cs reported agreed almost exactly with those obtained with the single channel detector described above. Calculations of counting time intervals corre- sponding to various counting rates were made using the following formulae (Overman and Clark, 1960): T = Rs+b + '/(Rb) (R660) 3 (GZ) where: TS = Sample time (min) R8 = Sample rate (min) Rb = Bkgd rate (min) Rs+b = Total rate (min) G = % Error (0.05) 98 R where: Tb = Bkgd time (min) Thus, each 137Cs determination is significant at the 95% level. Cs analysis was accomplished by means of flame emission spectrophotometry (Jarrell-Ash). _The principle emission line of Cs (8521 i) was utilized. The detection limit was about 0.1 ug/ml, the sensitivity was about 0.2 ug/ml. Cs standard solutions were prepared in 1.0 N NaOH solution and extracted into TPB solution, similar to fish samples. Recorder peak heights of standards were measured and the values were integrated by regression analysis to form a linear relationship between standard concentrations and response. Peak heights of samples were then applied to the regression equation to obtain appropriate values of Cs concentration. Test blanks were treated exactly like fish solu- tions and analyzed for Cs to detect background contributions of Cs from reagents (Table C-S). None of the blanks yielded positive results for Cs. To date, total Cs analyses have not been accom- plished for lake water samples. Consequently, concentra- tions reported in the data are estimates calculated from the relationship: 99 137 137 Cs in fish = Cs in H20 Cs in fish Cs in H20 ' The equality of the specific activities of Cs between fish and their medium has been demonstrated by several workers: Nelson, 1967; Folsom, et al., 1967; and Palmer and Beasley, 1967. ' APPENDIX D DATA TABLES APPENDIX D DATA TABLES TABLE D-1.--Physica1 characteristics and Cs levels in fish collected in Wintergreen Lake. Collection :33: Weight Length Sex Age 137C3 3::ii.€s Date # (gms) (cm) (yrS) (pm/gm) (mg/gm) 6-5-67 A 1047 42.5 F 5 B 697 35.5 F 4 0.68 C 384 31.4 M 3 D 322 30.5 M 3 0.67 7-3-68 4 500 32.5 F 3 0.69 6.76 6 580 34.5 M 3 0.87 7 470 34.0 F 3 1.10 8 610 35.5 M 3 1.08 7.35 9 790 38.0 M 4 1.14 11.54 10 600 36.0 F 3 1.04 11.11 11 750 38.5 F 3 0.85 8.60 12 310 31.0 M 3 0.83 13 800 39.0 F 3 1.04 14 380 31.0 M 3 0.66 8.23 15 950 40.0 M 4 0.87 16 600 35.5 M 3 0.69 17 540 35.5 M 3 0.77 19 630 35.0 F 3 0.97 20 540 34.5 F 3 0.63 21 500 34.0 M 3 0.76 22 280 27.0 F 2 0.50 24 550 34.0 M 3 0.70 5-19-69 25 365 30.0 M 3 0.46 28 295 29.4 M 3 0.48 29 280 29.2 M 3 0.56 3.75 30 170 24.2 I 2 0.26 5 73 31 185 25.2 I 2 100 101 TABLE D-1.--Continued Collection 2:22 Weight Length Sex Age 137C8 ¥ZE::.$S Date # (gms) (cm) (yrs) (pCi/gm) (ng/gm) 5-19-69 32 210 26.2 M 2 0.28 33 230 27.0 M 2 0.22 3.20 34 345 30.3 F 3 0.44 35 410 33.0 F 3 0.71 5.22 36 360 30.5 M 3 0.35 5.93 37 420 32.5 M 3 0.42 38 440 33.0 F 3 0.54 39 340 30.4 M 3 0.54 40 430 31.6 M 3 0.43 4.41 41 380 31.4 M 3 0.50 4.82 42 550 35.8 M 3 0.58 5.92 43 530 34.9 M 3 0.51 8.44. 44 520 34.8 F 3 0.59 5.23 45 630 36.8 F 3 0.48 4.89 46 670 37.0 M 3 0.58 9.43 47 640 36.5 M 3 0.54 48 670 36.5 M 3 0.80 12.09 49 580 36.6 M 3 0.63 10.18 50 740 38.3 F 3 0.80 8.32 51 1180 42.5 F 4 0.79 7.15 52 1360 43.7 F 4 0.70 5.01 53 1090 41.6 M 4 0.91 4.96 54 1300 43.2 M 4 0.58 8.98 6-20-69 55 450 34.0 M 3 0.66 60 350 31.9 F 3 0.43 10.90 61 350 30.8 F 3 0.39 12.17 62 350 30.5 F 3 0.37 10.83 63 300 29.3 M 3 0.34 16.14 65 300 28.9 F 3 0.29 16.95 7-17-69 68 320 28.0 F 3 0.31 15.13 69 380 30.5 M 3 0.37 13.66 70 350 30.5 M 3 0.38 15.21 71 370 29.5 M 3 0.33 15.13 72 320 28.0 M 3 0.35 15.22 8-12-69 73 350 29.0 M 3 0.25 12.31 76 435 31.0 M 3 0.22 11.69 77 445 31.5 M 3 0.31 12.93 78 440 32.0 M 3 0.30 15.75 79 370 30.0 M 3 0.31 12.93 102 TABLE D-l.--Continued Collection :33: Weight Length Sex Age 137Cs 322:: $8 Date # (gms) (cm) (yrs)(pCi/gm)(ng/gm) 9-18-69 80 620 36.5 M 3 0.36 14.65 81 530 33.5 M 3 0.24 9.15 82 490 32.5 M 3 0.34 13.01 83 480 32.5 M 3 0.39 14.62 86 570 35.5 F 3 0.50 11.60 10-17-69 87 695 37.5 F 3 0.42 10.10 88 650 35.0 M 3 0.39 10.97 89 500 33.0 M 3 0.39 13.13 90 355 30.0 M 3 0.30 11.51 6-3-68 Sunfish 310(2) -— -- -- 0.19 Perch 181(2) -- —— -- 1.67 6-20-69 Sunfish 865(4) -- -- -- 0.13 Perch 299(2) -- -- -— 1.14 103 TABLE D-2.-—Physica1 characteristics and Cs levels in bass collected in Fine Lake. Fish 137 Total Cs Collection Weight Length Age Cs Code Sex . (corr.) Date # . (gms) (cm) (yrs)(pCi/gm)(ng/gm) 7-18-69 1 86 21.0 M 2 0.77 24.66 2 50 16.5 F 2 0.68 19.29 3 40 16.0 M 2 0.63 19.29 5 160 25.0 M 3 0.76 -- 6-28-69 9 410 33.0 M 4 0.57 10.67 10 330 31.0 M 3 0.58 17.33 11 270 28.0 F 3 0.49 16.80 14 330 28.5 F 3 0.52 16.29 15 275 28.0 F 3 0.66 16.13 16 215 26.0 M 3 0.74 17.73 104 TABLE D-3.--Physica1 characteristics and Cs levels in bass collected in Fair Lake. Collection F18h Weight Length Age 137Cs Total CS Date C°de (9%) (cm) sex (yrS) (pCi/gm) (con: ' ) # (ng/gm) 9-5-68 4 400 32.0 M 5 1.53 -- . 5 180 25.0 F 3 1.63 17.68 6 100 21.0 F 2 1.09 17.70 7 130 23.5 M 3 1.00 17.56 10-3-68 8 155 24.5 3 1.52 17.74 9 176 26.0 3 1.51 17.82 11 142 24.0 3 1.35 17.82 6-3-69 13 140 22.5 F 3 0.91 17.95 14 140 22.5 F 3 1.10 15.66 15 120 20.0 F 3 1.01 14.72 17 120 20.5 M 3 1.10 17.84 18 110 19.8 M 3 0.89 17.30 19 140 22.6 M 3 1.33 16.17 20 150 23.5 F 3 1.42 17.93 105 TABLE D-4.--Physica1 characteristics and Cs levels in bass collected in Dewey Lake. Collection 2:2: Weight Length Sex Age 137Cs 322:: $8 Date # (came) (cm) (yr8) (pCi/gm) (rig/91;”- 6-12-69 3 170 24.0 F 3 0.62 4 160 24.0 F 3 0.66 13.91 6 930 40.5 F 6 0.66 11.75 7 180 23.5 M 3 0.69 15.48 8 160 23.5 F 3 0.74 15.99 9 150 22.5 F 3 0.72 16.47 10 190 24.5 M 3 0.69 16.14 11 140 24.0 M 3 0.87 19.68 106 TABLE D-5.--Physica1 characteristics and Cs levels in bass collected in Cable Lake. Collection €33: Weight Length Sex Age 13?CS ?2:::.?8 Date # (gms) (cm) (yrs)(pCi/gm)(ng/gm) 9-68 1 350 31.0 3 1.26 23.47 2 188 25.0 3 1.30 3 274 30.5 3 1.27 4 245 29.0 3 0.73 5 235 28.5 3 1.14 6 240 29.0 3 0.85 6-12-69 13 200 26.0 M 3 1.19 12.45 14 295 29.0 F 3 0.49 11.95 15 285 29.0 M 3 0.65 11.83 16 170 24.5 M 3 1.07 21.26 17 185 25.5 F 3 0.82 24.11 18 210 26.5 F 3 1.06 11.83 19 170 25.0 F 2 0.64 21.77 107 TABLE D-6.--Physica1 characteristics and Cs levels in bass collected in Schnable Lake. Collection Fish Weight Length Age 137Cs Total CS Date Code (gms) (cm) Sex (yrs)(pCi/gm)(c°rr') # (mg/gm ) 9-6-68 1 420 31.5 F 4 0.13 5.98 6-19-69 2 270 24.0 F 4 0.10 7.45 3 130 20.0 M 3 0.10 12.17 4 220 24.5 M 3 0.09 8.56 5 180 24.5 M 3 0.10 13.03 6 220 25.0 F 3 0.10 12.78 7 150 23.0 M 3 0.09 12.59 8 160 22.0 M 3 0.11 12.17