PB83-254300
Influence of Diet aria Body Lipids on the
Bioconcentration of Endrin from Water in the
Fathead Minnow ('pimephales promelas')
Goeteborg Univ. (Sweden)
Prepared for
Environmental Research Lab.-Duluth, MN
Aug 83
U.S. Department of Commerce
National Technical Information Service
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nuguai, xjuj
INFLUENCE OP DIET AND BODY LIPIDS ON THE BIOCONCENTRATION OF ENDRIN
FROM MATER IN THE FATHEAD MINNOW (Plmephales promelas)
Goran Dave
Department of Zoopliyslology
University of Goteborg
Box 250 59
400 31 Goteborg, SWEDEN
Patricia Koslan
University of Wisconsin
Center for Lake Superior Envlronmantal Studies
Superior, Wisconsin
Grant No- CR 806860-01
Project Officer
James M. McKlra
U.S. Environmental Protection Agency
Environmental Research Laboratory-Duluth
Huluth, Minnesota 55804
This study was conducted in cooperation with the National Swedish Environment
Protection Board, Stockholm, Sweden, and the U.S. Environmental Protection
Agency, Environmental Research Laborstory-Duuth, Duluth, Minnesota 55804.
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rf FtgHNItTAL HtrtJMI UAIA
T ' fPteau read Immicttom on tkt rtvtrt* be/art computing)
1. REPORT NO. 3.
KPA-600/3-83-077
3, RECIPIENT'S ACCESSION NO.
PB8 5 254300
4. TITLE AND SUBTITLE
Influence of Diet and Body Lipids on the Bioconcentration
of Endrin from Water in the Fathead Minnow (Pimephales
pronelas)
S. REPORT DATE
August 1983
«. PERFORMING ORGANIZATION COOK
7. AUTHORIS)
Goran Dave and Patricia Kosian
S. PEnPORMINO ORGANIZATION REPORT NO.
i. PERFORMING ORGANIZATION NAME ANO ADORES*
U.S. Environmental Protection Agency
Environmental Research Laborstory-Duluth
6201 Congdon Boulevard
Duluth, Minnesota 55804
10. PROGRAM ELEMENT NO.
*ACTMKXhT n6.
CR 806860-01
12, SPONSORING AGENCY NAME ANO AOORESS
Same as above
-.3. TYPE OP REPORT ANO PERIOD COVERED
1*. SPONSORING AGENCY CODE
EPA/600/03
IB. SUPPLEMENTARY NOTES
i4» A
Th» mirmu of this studv was 1® ouantltv Hi# Importance of me fathead minnow's (Plmephales promelas) body
I Ipld content and Its cornposltlon In the bloconcentratIon of a lipophilic chemical (andrln) from water.
For three months prior to exposure, six groups of fish were fed reference research diets containing 0, to,
15 or 20f (dry weight diet bails) lipids added as corn oil and/or salmon oil, Two other groups were fed frozen
brlno shrimp lArtecnla sol Ins) at two ratton levels.
BloconcentratIon tests at two concentrations of endrin In water {0,11 and 0.19 pg L~') produced mean
btoconcantrat1on factors (BCFs) of 15,000* after 14 days and 13,000* after It days whan expressed on a wet
weight, whole body basis* Corresponding naan SCFs expressed on a lipid, whole body basis mr& 190,000* and
340»Q00x.
Whole body 0CFs expressed on a wet weight basis ranged 8,000x - 2tr0Q0x after 14 days exposure and 5,000x -
30,000* after 29 days exposure. Independent of diet composition, whole body BCFs expressed on a wet weight
basis were positively correlated to the concentration of total fish body lipids. When BCFs were expressed on a
lipid basis, they were Instead negatively correlated to the concentration of total fish body lipids. From the
limited nunbar of samples examined for each diet group, no Influence of diet lipid source (corn oil, salmon oil
and brine shrimp lipids} could be found.
Results from this study were Interpreted as follows! the greater the fishes' body lipid content the more
andrln It Bloconcentrates rapidly and directly from the water, thus, fatter fish take a longer time to reach
equt1IbrluM.
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U.S. Environmental Protection Agency policy and
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or commercial products does not constitute endorse-
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ii
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ABSTRACT
The purpose of this study was to quantify the importance of the fathead
minnow's (Plmephales proraclas) body lipid content and its composition in the
bioconcentration of a lipophilic chemical (endrin) from water.
For three months prior to exposure, six groups of fish were fed
reference research diets containing 0, 10, 15 or 20% (dry weight diet basis)
lipids added as corn oil and/or salmon oil. Two other groups were fed frozen
brine shrimp (Artewla sallna) at two ration levels,
Bioconcentration tests at two concentrations of endrin in water (0.11
and 0.19 pg L-*) produced mean bioconcentration factors (BCFs) of 15,000x
after 14 days and 23,000x after 29 days when expressed on a wet weight, whole
body basis. Corresponding mean BCFs expressed on a lipid, whole body basis
were 190,000x jmd 340,000*.
Whole body BCFs expressed on a wet weight basis ranged 8,Q00x - 21,000x
after 14 days exposure and 5,000x - 3Q,000x after 29 days exposure.
Independent of diet composition, whole body BCFs expressed on a wet weight
basis were positively correlated to the concentration of total fish body
lipids. When BCFs were expressed on a lipid basis, they were instead
negatively correlated to the concentration of total fish body lipids. From
the limited number of samples examined for each diet group, no influence of
diet lipid source (corn oil, salmon oil and brine shrimp lipids) could be
found.
Results from this study were interpreted as follows: the greater the
fishes' body lipid content to more endrin it bioconcentrates rapidly and
directly from the water. Thus, such fish take a longer time to reach
equilibrium.
iii
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CONTENTS
Abstract iii
Figures ..... ........ v
Tables ..... . ......... vi
Acknowledgment .... ........... vii
Introduction ..... . ....... 1
Conclusions 2
Recomendat ions 3
Materials and Methods ....... .... 4
Experimental Procedures ........... 6
Results ................... . 8
Discussion 10
References 12
Appendix
Analytical data from endrin residue and lipid content analysis . . 24
iv
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FIGURES
Number Page
1 Correlation between mean total body water and mean total
body lipid contents for eight groups of fathead minnows
fed different diets for at least three months ~ . « 21
2 Correlation between bioconcentration factors (BCFs) for
endrin from water and total body lipid contents of
fathead minnows after 14 and 29 days exposure when
expressed on a wet weight basis .......22
3 Correlation between bioconcentration factors (BCFs) for
endrin from water and total body lipid contents of
fathead minnows after 14 and 29 days exposure when
expressed on a lipid basis 23
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TABLES
Humber Page
1 Gross Composition of Experimental Diets 14
2 Concentrations of Endrin in water (pg L~*) During
Exposure of Fathead Minnows ....>.15
3 Bioconcencration Factors {BCFs) for Endrin after 14 Bays
Exposure through Hater in Fathead Minnows Fed Different
Diets 16
4 Bioconcentration Factors (BCFs) for Endrin after 29 Days
Exposure through Mater in Fathead Minnows Fed Different
Diets 17
5 Lipid and Water Content of Fathead Minnows Fed Six
Artificial Diets with Increasing Lipid Contents or Frozen
Brine Shrimp at Two Ration Levels 18
6 Mean Bioconcentration Factors (BCFs) of Endrin through Water
in Fathead Minnows Fed Different Diets . 19
7 Linear Regression Analysis 20
vi
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ACKNOWLEDGMENTS
This work was performed at the Environmental Research Laboratory-Duluth,
Duluth, Minnesota. The cooperation of Its entire staff is gratefully
acknowledged. We are particularly indebted to Dr. Janes McKlm, Chief of the
Physiological Effects Section, and all the Members of this section.
Chemical analysis of water samples were carried out by Mrs. Diane Olson
and Mr. Steve Peterson.
Dr. David Weininger and Mr. Paul Telega provided their help for the
statistical treatment of the data.
Financial support was provided by the Sweden-America Foundation, the
National Swedish Environment Protection Board (Project Number 7-338-78), and
the United States Environmental Protection Agency (Grant No, R806860-01).
vii
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INTRODUCTION
In aquatic animals bioconcentration of chemicals can take place directly
from water and from food. When Jarvinen and Tyo (1978) exposed fathead
minnows (Ploephalea pronelas) to endrin, both through water and through food
(consisting of previously exposed clans) for 300 days, they found maximum
bioconcentration factors (ICFs) of 0.8x from food and 13,000x from water,
the conclusion from their study is that the uptake from water was the
predominant route for endrin in the fathead minnow.
Prediction of the bioconcentration factor, BCF (¦» concentration in fish
divided by concentration in water at steady state), from the pertition
coefficient between octanol and water (P) has been thoroughly Investigated
(Neely et al., 1974} Veith et al., 1979; Koneraann, 1979; Renberg and
Sundstron, 1979). In these studies the general equation: Log BCF ¦ aQ +
a^ x Log P has given good correlations for a wide variety of chemicals.
However, in estimates made on the same product, factors like source of fish
and/or experimental error, species and test temperature have produced
variations in BCF (for Aroclor 1016) of 2, 4 and 12 times, respectively
(Veith et al., 1979).
The purpose of this study was to estimate the relative importance of
some nutritional factors on the fathead minnow's ability to bioconcentrate a
lipophilic chemical, endrin, from water. The nutritional factors studied
were dietary lipid content and composition and ration level. The influence
of these factors and of starvation on the toxicity of endrin has been
reported in a simultaneous study (Dave, 1981).
1
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CONCLUSIONS
The results from this study have shown that the body lipid content had a
significant effect oil the bioconcentration of a lipophilic chemical, endrin,
in the fathead minnow. The fatter the fish was, the more endrin it
bioconcentrated directly from water. Bioconcentration factors (BCFs) after
29 days exposure ranged 5,OOQx - 30,000x (6-fold difference), when expressed
on a whole body wet weight basis. When instead expressed on a lipid basis,
BCFs after 29 days expojura ranged 250,000x - 450,OOOx (2-fold difference).
BCFs calculated on a wet weight basis were positively correlated to the
lipid content of the fish, but BCFs calculated on a lipid basis were instead
negatively correlated to the lipid content, Indicating that the fatter the
fish was, the longer was the tine needed to reach a steady state.
2
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RECOMMENDATIONS
Bioconcencretion factors (BCFs) for lipophilic chemicals should be
expressed on a lipid as well as a wet weight basis. Since BCFs for
lipophilic chemicals like endrin estimated in 30-diy bioconcentration tests
can be expected to be significantly affected by the lipid content of the
fish. A fatter fish is expected t,o show* a higher residue than a lean fish.
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MATERIALS AND METHODS
A detailed description of the materials and methods used have been given
previously (Dave, 1981). Therefore, only a brief presentation of the
experimental design and the method for residue analysis is provided here.
One-month-old fathead minnows were separated into eight groups (I-VIII),
which were kept in separate tanks for three months prior to exposure to
endrin. Diets fed to group I-VI were reference research diets (Brauhn and
Schoettger, 1975; National Research Council, 1973; 1977) containing different
percentages or sources of lipids. Group Vll and VIII were fed a high (ad
lib) and a low (1/6 of ad lib) ration of frozen brine Bftrimp, respectively.
Met compositions are given in Table 1.
After this feeding period, exposure to endrin through water was perform-
ed simultaneously for all groups by enclosing them in screened compartments
in a flow-through exposure system. Water temperature was 25.1+0.7°C
(mean+S.D.) . Alkalinity ranged 39.5-41.0 mg L"1 as CaCC>3, hardness
43.5-45.5 mg L~^ as CaCOj, and pH ranged 7.3-7.5 as determined weekly
on the laboratory source (take Superior water). Calculated mean endrin
concentrations in water based on chemical analyses are given in Table 2.
For the residue analysis composite, whole fish samples were homogenized
with 70 g of granular anhydrous ^2^04 (Mallinckrodt, Inc.). The
powdered homogenate was transferred to a 300-ml chromatographic column and
eluted with 250 ml of redistilled hexane. The eluate was collected in a
250-ml volumetric flask. No cleanup procedure was performed. A 10-ml
aliquot of the sample was transferred to a culture tube and stored in the
freezer to await analysis by gas chromatography. The remaining 240 ml were
used for lipid content measurements.
4
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The 240-ml samples were transferred to 250-ml beakers and concentrated
to 10 ml. Clean, solvent-rinsed 25-ml beakers were heated to 110'C for 30
minutes, cooled in a desiccator for 30 minutes and weighed to four decimal
places. The samples were then quantitatively transferred with methylene
chloride to the tared beakers and allowed to evaporate to near dryness. The
beakers were then heated to 110'C for 30 minutes, cooled in a desiccator for
30 minutes and reweighed to four decimal places. The lipid content was based
on the following calculation:
, . . . , (gross - tare) x 100
(Pe cetr ^ " tissue weight x 0.96
The .scored 10-ml samples were re-adjusted to volume and screened on the
gas chromatograph to determine the proper dilution ratio. After necessary
dilutions were made, 1.5-ml aliquots of the samples were transferred to
Hewlett-Packard injection vials for quantitation by gas chromatography.
The gas chromatographic analysis was performed on a 573QA
Hewlett-Packard gas chromatograph with an auto sampler and a Hewlett-Packard
3354B lab automation data system. The gas chromatograph was equipped with a
6%i electron capture detector held at 300°C, and the injection port
temperature was 250°C. The 6 ft. 2 x 3 mm (0D) column was packed with 1.5%
SP-2250/1.95X SP-2410 on 100/120 mesh Supelcoport. The carrier gas was 5%
methane in argon and a flow rate of 40 ml/min. The tissue samples were
analyzed at a column temperature of 210°C. The percent recovery of spiked
samples was 99.9+2.3 percent with n - 3, Results crom the analyzes are given
in the Appendix.
5
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EXPERIMENTAL PROCEDURES
The exposure to endrIn was started with 10 fish from each diet group
(I-VIII). The three highest concentrations (called 1, 2 and 3) delivered
from the diluter (Mount and Brungs, 1967) were used to measure toxicity,
concentrations 4 and 5 were used to measure bioeoncentration. Concentration
6 was the control (conc. » nil). Fishes for residue analyses were sampled
after 14 and 29 days. They were killed by immersion in 0*C water for 30
seconds, weighed to the nearest 0.01 g and deep-frozen in glass liquid
scintillation vessels to await residue analysis.
During the exposure period, mortalities of fish were 4/80 In cone. 4;
5/80 in conc. 5 and 1/80 in the control. Only live fishes were used for
residue analysis.
No food was given for the first 10 days of exposure or for 24 hr prior
to sampling. The other days, the previously fed diets were given at an
average ration of 2.8 g dry food/100 g live fish body weight per day to
groups 1-VII and 1.5 g dry food/100 g live fish body weight per day to group
VIII. For the entire exposure periods (14 and 29 days) the calculated
average daily ration for the first 14 days was 0.6 g dry food/100 g fish in
group 1-VII and 0.3 g dry food/100 g fish in group VIII, and for the 29 days
exposure corresponding figures are 1.6 and 0.9. Dally rations were corrected
for the reduced number of fish due to sampling after 14 days (4-5 fish in
each group) but not for mortalities. Calculated rations above are based on
measured amounts of food and calculated averages for body weights of sampled
fish (given in the Appendix).
Because of the errors in feeding a group as opposed to individual fishes
and the variations in fish size within and between the different groups, no
6
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detailed analysis of body weight gain or loss during exposure seemed valid.
The diet type provided during the three months prior to and during exposure
was the same and the over-all ration given during exposure was approximately
a maintenance ration.
7
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RESULTS
Mean concentrations of endrin in water during exposure for 14 and 29
days are given In Table 2, These concentrations and corresponding whole body
residues of endrin and whole body lipid concentrations given in the Appendix
were used for calculations of BCFs (bioconcentration factors)« BCFs
calculated both on a wet weight and on a lipid basis are given in Table 3 for
14 days exposure and In Table 4 for 29 days exposure•
Hean values for whole body lipid contents (from Appendix) and whole body
water contents (from Dave, 1981) for groups I-VII1 are given in Table 5.
Hean lipid contents (Table 5) and analytical data in the Appendix suggest
that individual variations In whole body lipids among these fish are too
great to show a direct relationship to dietary lipid content. However,
independent of dietary lipid there is a correlation between the mean whole
body lipid contents for the different groups of fish (I-VIII) and their whole
body water contents (shown in Figure 1). This relationship between lipids
and water has been repo ted for several other fish species (review by Love,
1970). The present study shows that such a relationship, the fat-water line,
also exists in the fathead minnow.
Because of the great individual variations in lipid contents as well as
in endrin residues, the data were tested for possible correlations between
lipid contents and BCFs (independent of dietary treatment). In Figure 2 we
have presented the linear regression of BCFs expressed on wet weight basis
and whole body lipid concentrations for the samples from all diet groups.
The values for aQ (lipid content = 0) after 14 and 29 days are almost
identical (7, 516 and 7,423), but the calculated value for a^ for 29 days
exposure is more than twice that for 14 days exposure (2,269 and 908,
8
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respectively). These significant correlations must mean, that the fattier
the fish is, the more endrin it bioconcentrates directly from water.
Furthermore, the higher value for after 29 compared to 14 days exposure
oust mean, that the fattier the fish is, the longer is the time required to
reach equilibrium.
In Figure 3 we have presented the linear regression of BCFs expressed on
a lipid basis and whole body lipid concentrations for the samples from all
diet groups. The inverse relationships suggest, that the fattier the fish
is, the less saturated is its lipid pool with endrin after identical
exposures. Or in other words, the fattier the fish is, the longer the time
it takes to reach equilibrium.
The source of lipid in the diet is of nutritional importance because it
determined the dietary fatty acid composition and the proportions of
triglycerides, cholesterol, phospholipids, etc. The lipid sources used in
the present study were corn oil, which is high in fatty acids, salmon oil,
which is high in Jl3 fatty acids, and the brine shrimp lipids (Artemla
sallna, San Fransisco Bay variety, analyzed by Gallagher and Brown, 1975).
The present study did not indicate that the source of dietary lipid had any
influence on the results obtained. However, it might have affected the lipid
composition of the experimental fish, but this was not Investigated.
9
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DISCUSSION
The results from this study have shown that the body lipid content had a
significant effect on the bioconcentration of a lipophilic chemical, endrin,
in the fathead minnow. The fattier the fish was, the more endrin it
bioconcentrated directly from water. Bioconcentration factors (BCFs) after
29 days exposure ranged 5.000* -30.000x (6-fold difference), when expressed
on a whole body, wet weight basis. When instead expressed on a lipid basis,
BCFs after 29 days exposure ranged 25Q.OOOx - 450.000x (2-fold difference).
Among different species of fish, there are considerable variations in
body fat deposit distributions, total amount and composition. Furthermore,
there are variations within a species caused by age, availability of food,
season, sex, size and strain (review by Love, 1970). Considering these
variations, estimates of BCFs for lipophilic chemicals solely on a wet weight
basis can be expected to produce considerable variations. Estimates of BCFs
both on a wet weight and a lipid basis would contribute to a better
understanding of causes for variations in bioconcentration within the same
species and perhaps to some extent also between different species of fish.
The results from this study reveal a positive correlation between BCFs
expressed on a wet weight basis and total body lipid contents, indicating a
higher uptake from water in fat compared to lean fishes. When BCFs were
expressed on a lipid basis, they were instead negatively correlated to the
total fish body lipid content, indicating that fattier fishes needed a longer
time than leaner fishes to reach equilibrium. This implies that estimates of
steady state BCFs would require different periods of exposure depending on
the total lipid pool- However, other species-related factors, like gill
surface area and detoxification capacity, are probably also of great
10
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importance for the time to reach equilibrium. Estimates of bioconcentration
in different species of fish under identical expoaures, would probably reveal
the relative importance of such species-related factors. Such comparative
estimates would probably also extend the possibilities to predict
bioconcentration of lipophilic chemicals in different species of fish and
under different physiological conditions related to age, nutritional status,
season and sex. Because of the importance of the lipid content in the
bioconcentration process, BCPs should be expressed both on a wet weight and a
lipid basis in such studies.
The present study dealt only with uptake directly from water. Under
natural conditions uptake of chemicals can take place also from food. From
an energetic point of view, a fat compared to a lean fish of the same age and
body weight and within the same population (environment and type of food
being the same) should have consumed aore food. Because of this greater food
consumption, a fattier fish under natural conditions would be expected to
have experienced a greater uptake both through food (due to bioenergetics)
and through water (due to partitioning).
11
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REFERENCES
Brauhn, J. 1».» and R. A. Schoettger. 1975. Acquisition and culture of
research fish: rainbow trout, fathead minnow, channel catfish and
bluegills. U.S. E.P.A., Ecological Research Series, EFA-660/3-75-011.
55 p»
Dave, G. 1981. Influence of diet and starvation on toxicity of endrio to
fathead minnows (Flraephales promelas). U.S. Environmental Protection
Agency, Duluth. Ecological Research Series (In Press).
Gallagher, M., and W. D. Brown, 1975. Composition of San Fransisco Bay
brine shrimp (Arteraia salina). J. Agric. Food Chen. 23: 630-633.
Jarvinen, A. W., and R. M. Tyo. 1978. Toxicity to fathead minnows of endrin
in food and water. Arch. Environ. Contain. Toxicol. 7: 409-421.
Konemann, W. -H. 1979. Quantitative structure-activity relationships for
kinetics and toxicity of aquatic pollutants and their mixtures in fish.
Ph.D. Dissertation. University of Utrecht, The Netherlands. 79 p.
Love, R. M. 1970. The Chemical Biology of Fishes. Academic Press, New
York. 547 p.
Mount, D. I., and W. A. Brungs. 1967. A simplified dosing apparatus for
fish toxicology studies. Water Res. 1: 21-29.
National Research Council. 1973. Nutrient requirements of domestic animals,
No. 11: Nutrient requirements of trout, salmon and catfish. National
Academy of Sciences, Washington, D.C. 57 p.
National Research Council. 1977. Nutrient requirements of domestic animals:
Nutrient requirements of warmwater fishes. National Academy of
Sciences, Washington, D.C. 78 p.
12
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Neely, W. B., D. R« Branson, and G. E. Blau. 1974. Partition coefficients
to measure bloconcentratlon potentials of organic chemicals In fish*
Environ. Sci. Technol. 81 1113-1115.
Renberg, L., and C# Smdstroia. 1979. Prediction of bloconcentratlon
potential of organic compounds using partition coefficients derived from
reversed phase thin layer chromatography. Chemosphere 7; 449-459.
Veith, G. D.» D» L. DeFoe, and B. W. Bergstedt. 1979. Measuring and
estimating the bloconcentratlon factor of chemicals in fish. J. Fish.
Res. Board Can. 36: 1040-1048.
13
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Table 1. Cross Composition of Experimental Diets
Ingredient
Amount of Ingredient (g) in Respective Diet
II
III
IV
VI *
Casein
280
280
280
280
280
280
Gelatin
120
120
120
120
120
120
Dextrin
280
280
280
280
280
280
Vitamin mix.
10
10
10
10
10
10
Mineral mix*
40
m
40
40
40
40
Sum of above
730.
730
730
730
730
730
Com oil
-
ioo
-
50
75
100
Salmon oil
-
-
100
50
75
100
Sua of above
730
830
830
830
880
930
a~cellulose
230
170
170
170
120
70
Sun of above
1000
1000
1000
1000
1000
1000
Added water
2000
2000
2000
2000
2000
2000
Moisture (%)
66 <7
66.7
66.7
66.7
66.7
66.7
~Values for diets called VII and VIII, which were frozen brine
shrinp Artemis sallna (San Franslsco Bay Brand.;' Newark, California
94560; Stock #65006) as determined by Gallagher and Brown (1975)
were as follows: protein 58% dry weight (Kjeldahl method), crude
fat 5.IX drv weight (ether extract) or .19.3% dry weight (methanol-
chloroform cictract), fiber 3.5% dry weight, ash 20.6% dry weight,
at 901 moisture.
14
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Table 2. Concentrations of Endrin in Mater (pg L"*^) During
Exposure of Fathead Minnows
Period of Exposure
Cone. 4*
Gone. 5
Day Q-.lb
Day 0-29
0.18392**
<0.13277 - 0.23506)***
0.19674
(0.14370 - 0.24578)
0.10829
(0.09158 - 0.13e40)
0.11466
(0.08461 - 0.14471)
* Cone. 4 and 5 were the two lowest concentrations delivered by the
diluter (Mount and Brungs, 1967).
** Mean values calculated from analytical data of regular samples
taken from the dipping bird reservoir and exposure tanks. The
analytical procedure does only justify two numbers, e.g., 0.18 jig
L~ . Calculated mean value* in the table have been used for
calculations of BCFs.
*** Number* in parenthesis are 95% confidence limits.
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Table 3. Bioconcentration Factors
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Table 4. Bioconcentration Factors (BCFs) for Endrin After 29 Days Exposure
Through Water in Fathead Minnr>*tf"Fed Different Diets
Endrin in Water Total Body Lipid BCFi4 days
Diet Group pg L g/100 g W.P. Basis Lipid Basic,
I
0.19
{cone. 4)
1.34
5,068
378,248
11
0.19
(cone. 4)
9.90
24,669
249,204
III
0.19
(conc. 4)
5.39
20,782
385,540
IV
0.19
(conc. 4)
8.64
24,653
285,355
¥
0.19
(conc. 4)
10.07
25,644
254,647
VI
0.19
(conc. 4)
7.99
27,760
347,43d
VII
0.19
(conc. 4)
6.41 -
28,654
447,006
VIII
0.19
(conc. 4)
28
24,423
299,733
I
0.11
(conc. 5)
2.69*
10,291
382,565*
11
0,11
(conc. 5)
7.04
22,100
313,885
III
0.11
(conc. 5)
•J. 96
30,167
411,174
IV y*
0.11
(conc. 5)
7,23
20,487
283,360
V
0.11
(conc. 5)
9.29
30,167
324,699
VI
0.11
(conc. 5}
8.91
25,301
283,970
VII
0.11
(conc. 5)
8.20
30,054
366,475
VIII
0.11
(conc. 5)
3.71
12,666
368,394
*Mean lipid value for diet group I and BCF calculated from this value, because
lipid value was missing. ^
17
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fable 5. lipid And Water Content of Fathead Minnows Fed Six Artificial Diets
With Increasing Lipid Contents or Frozen Brine Shrimp at Two Ration Levels
Diet
Fish Body,
Mean + S.O. (n)
No.
Lipids*
Hation
Lipids (X)
Water (X)
I
no lipids
unrestricted
2.69+0.88
(4)
78,8+2.8 (8)
II
10% C.O.
unrestricted
8,99+1.37
C5)
73.3+2.1 (8)
III
10% s.o.
unrestricted
7.06+1.92
<5>
73.5+1.5 (8)
IV
5% C.O, + 5% S.O.
unrestricted
8.80+1.42
(5)
70.1+2.3 (8)
V
7.5% C.O. + 7.5% S.O.
unrestricted
9-24+0-85
(5)
73.1+2.6 (8)
VI
10% C.O. + 10% S.O.
unrestricted
10.33+1.75
(5)
71.6+2.6 <8)
VII
Brine shrimp lipids
unrestricted
10.13+3.01
(5)
68,4+2.9 (8)
VIII
Brine shrimp lipids
restricted
5.58+1.42
(5)
73.3+2.5 (8)
ca 1/6 of VII
*C,G. ¦ corn oil; 5.0. =» salmon oil. Lipid percentages are nominal values on dry
weight basis.
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Table 6. Mean BioconcentraCiwi Factors (BCFs) of Endrin Through Water in Fathead
Minnows Fed Different Diets
Endrln Concentration, us L**1-
Period of 0,18 - 0.19 (cone. 4) _ 0.11 (conc. 5)
Exposure BCF W.W. Basis BCF Lipid Basis 1CF W.W. Basis BCF Lipid Basis
14 days 16,266* 210,017 14,487 173,158
+3,886 (24%) +49,150 <23%) +3.084 (21%) +39,618 (23%)
29 days 22,707 330,896 22,779 341,815
+7,511 (33%) +70,220 (21%) +7,683 (34%) +47,241 (14%)
*BCFs given as mean + S.D. (n - 8) with C.V. (S.D. / mean x 100) in parenthesis. Only
two to three significant figures are justified.
19
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table 7. Linear Regression Analysis*
Y
X
a0
al
r
N
Value
of P
S.D.
Data Presentatior
BCF after
14 days (w.w. basis)
Body lipid (X)
7,516
908
0.815
16
0.001
2,105
Table 3; Fig. 2
29 days (w.w. basis)
Body lipid CI)
7,423
2,269
0.795
16
0.001
4,606
Table 4; fig. 2
14 days (lipid basis)
Body lipid (2)
303,292
-12,909
0.863
16
0.001
24,652
Table 3; Fig. 3
29 days (lipid basis)
Body lipid (%)
430,238
-13,902
0.616
16
0.01
47,372
Table 4; Fig. 3
Mean body water {%)
Mean body
lipid (%)
80.51
-0.986
0.846
8
0.01
1,759
Table 5; Fig. 1
Endrin residue (ppm)
after
14 days in conc. 4
Lipid content (I)
1,375
0.1937
0.902
8
0.01
0.334
Appendix
14 days in conc. 5
Lipid content (Z)
0.763
0.0899
0.851
8
0.01
0.189
Appendix
29 days in conc- 4
Lipid content (X)
1,207
0.4370
0.833
8
0.02
0.911
Appendix
29 days in conc. 5
Lipid content (%)
0.433
0.3166
0.921
7
0.01
0.346
Appendix
*y ¦ iQ + a| x X
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Y» 80.5-0.99 X
4 "T1 I i
0 S 10
LIPID CONTENT, %
Figure 1. Correlation between mean total body water and mean
total body lipid contents for eight groups of fathead
minnows fed different diets Tor at least three months.
Roman numerals refer to diet numbers.
21
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LIPID CONTENT, %
Figure 2. Correlation between bioconcentration factors (BCFs)
for endrin from water and total body lipid contents
of fathead minnows after 14 and 29 days exposure when
expressed on a wet weight basis.
22
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Figure 3. Correlation between bioconcentration factors (BCFs)
for endrin from water and total body lipid contents
of fathead minnows after 1b and 29 days exposure when
expressed on a lipid basis.
23
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APPENDIX
ANALYTICAL DATA PROM ENDRIN RESIDUE A® LIPID CONTENT ANALYSIS
Lipid
Sample Wet Tissue Endrin (ppu) Content
Name Weight (go)
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APPENDIX (Continued)
Lipid
Sample Wet Tissue Endrin (ppn) Content
Name Weight (gm) Cpg/gm) (percent)
B1K1
<0.01
5/29
I5 6
0.27
0.771
-
5/29
I5 8
0.71
1.589
-
5/29
II5 6-9
2.54
2.534
7.04
5/29
III5 6-8
3.56
2.814
5.96
5/29
IV5 !j—10
3,81
2.349
7.23
5/29
V5 6-10
3.09
3.459
9.29
5/29
VI5 6-8
1.83
2.901
8.91
5/29
VII5 6
1.02
4.162
-
5/29
VII5 7
0.78
7.059
-
5/29
VII5 8-10
2.51
3.446
8.20
5/29
VIII5 6-9
2.01
1.567
3.71
B1K2
-
<0.01
-
5/29
I6 1-5
2.35
<0.01
2.50
5/29
II6 1-5
2.8ft
<0.01
9.03
5/29
III6 1-5
3.65
<0.01
9.80
5/29
IV6 1-5
4.62
0.159
7.89
5/29
V6 1-5
3.54
<0.01
9,82
5/29
VI6 1-5
3.43
<0.01
11.45
5/29
VII6 1-55
5.89
<0.01
9.76
5/29
VIII6 1-5
2.39
<0.01
7.33
Name of samples indicates/days of exposure, diet group (roman), endrin
concentration (actual concentrations in Table 2), numbers of fish in the
composite sample.
25
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