War. Res. Vol. 24, No. 3, pp. 321-327, 1990 0043-1354/90$3.00+ 0.

00
Printed in Great Britain.All rights reserved Copyright 1990PergamonPress plc

IN SITU DETERMINATION OF PCB CONGENER-SPECIFIC

FIRST ORDER ABSORPTION/DESORPTION RATE
CONSTANTS USING CHIRONOMUS TENTANS LARVAE
(INSECTA: DIPTERA: CHIRONOMIDAE)
M. A. NOVAK 1, A. A. REILLY 2, B. BUSH 2 and L. SHANE2
~New York State Department of Environmental Conservation, Division of Water, 50 Wolf Rd, Albany,
NY 12233 and 2New York State Department of Health, Wadsworth Center for Laboratories and
Research, Albany, NY 12201, U.S.A.

(First received May 1988; accepted in revised form September 1989)

Abstract--The uptake of polychlorinated biphenyl (PCB) congeners was measured in the larvae of a
laboratory-reared chironomid midge, Chironomus tentans, placed in the upper Hudson river, New York
during 2 months in 1985. This procedure was investigated as a method for determining water congener
concentrations during times of fluctuating PCB levels, and to model uptake of PCBs by river biota. After
a 96 h exposure period, total PCB concentrations in the test organisms averaged 6.7 #g g-t total PCBs,
compared with water concentrations of 67 ng 1- t (mean value for both months). Uptake and elimination
constants, time to equilibrium and concentration factors were calculated for each of 21 selectedcongeners.
Analysis of PCB congeners in insects harvested at intervals during the 96 h period showed that uptake
differs with varying degrees of chlorination relative to water concentrations. At the end of the exposure
period, concentration factors ranged from 4000 to over 300,000 times the water concentrations.
Differences in the replicate indicate potential problems with this method as a field tool; instead of using
all congeners separated in analysis, several individual congeners should be selected for use based on their
importance to the river fauna and the consistency with which they are analyzed.

Key words--PCB congeners, kinetic studies, Chironomus tentans, chlorinated xenobiotics, bio-uptake

INTRODUCTION The current article reports on a study conducted in

Several aquatic invertebrates, including freshwater
clams (Mollusca: Pelecypoda), chironomid midges
(Insecta: Diptera: Chironomidae) and hydropsychid
caddistties (Insecta: Trichoptera: Hydropsychida)
(Hartley and Johnston, 1983; Bush et al., 1985;
Simpson, unpublished results) have been used for
biological assessment of aquatic contaminants. These
organisms accumulate lipophilic compounds to
equilibrium concentrations in excess of those in their
environment.
Previous investigators have focused on either
laboratory determination of uptake kinetics in the
presence of constant contaminant concentration or
field determination of bioconcentration. Our goal has
been to estimate congener (isomer) specific kinetic
parameters from time sequence field measurements of
both water and organism concentrations. Aside from
their intrinsic biological utility, these parameters
provide the ability to predict uptake given only
waterbody measurements. For most compounds of
interest, the relationship between water column
concentrations and biological uptake has not been
investigated; we anticipate that considerable savings
can be realized through in situ determination of these
parameters by exposing uncontaminated individuals
to contaminated natural environments.
the Hudson river, New York, in July and September
1985, using third and fourth instar laboratory-
cultured Chironomus tentans (Insecta: Diptera:
Chironomidae) larvae. The sediments of the upper
Hudson river are contaminated with PCBs from
capacitor manufacturing operations which dis-
charged for approx. 30 years. These sediments
contaminate the water column. The purpose of the
study was to document accumulation rates of various
PCB congeners in the test organisms.
MATERIAI.,S AND METHODS
Field procedures
A laboratory culture of Chironomus tentans was main-
tained, with modifications, according to the methods of
Townsend et al. (1981). An attempt was made to eliminate
all sources of PCB contamination by using deionized water
and glass rearing tanks, and minimizing the use of plastics;
Tygon~ tubing was used where necessary.
The larval exposures occurred in the upper Hudson river's
Thompson Island pool, a reach extending from Fort
Edward (at Lock 7) to the dam at Thompson Island. The
water was 3 m in depth, and current speed, measured at
the exposure site, was 24 cm s -~ in July, and 56 cm s-t in
September. Water temperatures were 21.5C in July and
18C in September.
Larvae were placed in the river in Nitex monofilament
nylon screen "envelopes" (mesh opening 560 microns;
Tetko, Elmsford, New York), measuring 6.5x 12cm.

321
322 M. A. NOVAKet al.

Twenty-five third and fourth instar larvae were placed in
each cage, and transported to the field in hexane-rinsed
galvanized steel pails containing deionized water. The cages
were placed, in groups of ten, into steel mesh baskets, which
were suspended at a depth of 1 m, from floats anchored by
cinder blocks on the river bottom.
Triplicate samples were harvested at 0, I, 2, 4, 8, 12, 24,
48, 72 and 96 h. The cages were opened, larvae removed
with flexible forceps and counted. The insects were placed
in glass vials, the vials capped and put immediately on dry
ice. Controls (0 h) were caged and transported to the field,
then removed and frozen, without being put into the river.
Triplicate water samples were taken at each harvest.
Water was collected 0.3 m from the surface, in hexane-rinsed
21. jars fitted with Teflon cap liners. The jars were rinsed
three times with river water to equilibrate PCBs on the inner
surface, since these compounds adsorb to glass (Bush et al.,
1985). Samples were placed on ice for transport back to the
laboratory, where they were stored at 4C until analyzed.
Laboratory methods
Larval samples were lyophilized, ground with 2 ml n-hex-
ane in a Tekman Tissuemizer until reduced to a fine powder,
then extracted by the method of Bush and Barnard (1982).
Water samples were extracted with n-hexane, the extract
dried with sodium sulfate and concentrated to 1 ml. All
extracts were analyzed on a Hewlett-Packard 5840A gas
chromatograph equipped with a 5880 splitless glass capillary
inlet, and fitted with a 60m Apiezon L glass column
(0.25-0.3 mm i.d.); 76 congeners were separated by this
column (Bush et al., 1983). Quantities were calculated using
a reference peak compared to external standards of a known
congener mixture (200ngml -~ mixture of Aroclors 1221,
1016, 1254 and 1260; 1:1:1:1 in hexane). Recalibration was
performed after each set of six samples, by conducting an
analysis of the external standard, followed by a hexane sam-
ple to detect and remove any impurities left on the column.
Congener uptake
Twenty-one congeners were selected for study of uptake
dynamics during a 96-h exposure in the river. Accumulation
of ach PCB congener was modeled by utilizing the differen-
tial equation governing uptake dynamics:
dCt(t)
= -- K 2 CL(I ) + K, Cw(t ) (I)
dt
where
CL(t)=congener concentration in the larvae
(ng g- ~dry wt) at time = t
K~ = uptake rate constant (h-~)
K2 = elimination rate constant (h -~)
Cw(t) =congener concentration in water (ng 1-j)
at time = t.
Uptake of individual congeners cannot be modeled from
equation (1) unless the functional form of Cw(t), the time
specific river concentration, is known. Values for Cw(t) were
measured in triplicate at sampling times T~ = 0,
T2 = 1. . . . . Tt0 --- 96. Cw(t ) may therefore be approximated
with a sequence of linear functions:
Cw(t)=floi+fllit for Tj_I<t<T , (2)
where fl0~ and fll~ are the estimated intercept and slope
parameters for the ith interval.
Larval congener concentration at the harvest times,
CL(T~), may be modeled by integrating equation (l) utilizing
(2):
Ki F
CL(Ti) = CL(T i ) e-x2r, +_~,lflli(Ti_ T,_te-r2tr,-r,_,))
In equation (3), fl0~and flt~ are determined by Cw(T i_ t)
and Cw(T~), so that the only parameters to be estimated, as
indicated in equation (1), are K~ and /(2.
Water and C. tentans data were subjected to a weighted
classical non-linear least squares analysis utilizing equation
(3), with weights equal to observed replicate variances. The
lack of fit of each datum to the model was examined for
statistical significance (studentized residual greater than
2.3), and if found, the observation was deleted.
When Cw(t) is relatively constant, integration of equation
(1) results in:
Ce(t) = ~KI Cw(/)(1 - e -x2t) (4)
After long periods of exposure (t large) the last factor
approaches unity so that the expression for larval PCB
concentration (CL) becomes Kt/1(2 times the concentration
in the river. The time it takes to reach this approximate
equilibrium condition depends only on K2, the elimination
rate constant. The ratio of K~ to K: is considered the
concentration factor of a congener at equilibrium.
A practical estimate of equilibrium was defined as being
a value within 10% of the true value, as used by Sanders and
Chandler (1972). K~ and K2 values, concentration factors
and days to equilibrium (within 10%) were estimated for the
selected congeners. The number of days required to reach
90% equilibrium for each congener was estimated by setting
the last factor in equation (4) to 90%:
In 0.10 / 2 4 h
days _ K 2 h _ l / d a y (5)
RESULTS
P C B composition o f s a m p l e s - - J u l y
Total PCB c o n c e n t r a t i o n s in the July water
samples ranged from 47 to 166ng1-1 (n = 28 for all
10 exposure times) with a m e a n o f 93 ng 1-~ (Table 1).

Table 1. Total PCB concentrations in Chironomus tentans (ng g- ~dry wt) and
water (ngl-~), Hudson river, July and September 1985. Reported values are
means of 3 samples +- (rounded to 3 significant figures) 95% confidence limits
July September
Hour C. tenlans Water C. tentans Water
0 I11 + 124 95+ 19 1110+_69 66+-8
I 815+-677 61 +4 1310_+51 59+- 11
2 584+-508 72+- 11 1660+- 144 37+- 12
4 1200 -4-127 II0 +- 16 2120 + 72 31 + 40
8 1130 +- 429 113 +- 9 2600 _+ 203 36 +- 2
12 2440+- 1390" 115+- 13 3540+- 119 36+-9
24 2870 +- 1250 84 +- 6 5340 +- 924 38 +- 3
48 4020 +- 464 lOI +- 23 6640 +- 553 27 +- 2
72 3510+-794 66+-16 6480+-1550 38+-4
96 6160+- II00" 129 +-42 7250+- 1840 33+- I
*Mean of 2 values.
 Determination of PCB rate constants 323

T a b l e 2. P e r c e n t c o n t r i b u t i o n o f P C B c o n g e n e r s in r e p r e s e n t a t i v e Chironomus tentans ( a f t e r 8 a n d
4 8 h e x p o s u r e ) a n d w a t e r s a m p l e s ; J u l y a n d S e p t e m b e r 1985, U p p e r H u d s o n f i v e r , N e w Y o r k

C. tentans
Water 8h 48 h

Congener July September July September July September

2 -- -- -- 13 -- 5
2,2' 29 20 10 10 -- --
26 16 . . . . .
26,2' 32 11 15 6 -- --
25,2' 3 I1 . . . .
26,4' -- 9 . . . .
25,4' -- -- -- -- 5 5
24,4' -- -- 5 4 6 --
24,2'5' -- -- 5 -- 7 6
236,4' -- -- -- -- 7 6
All o t h e r
congeners 20 50 65 67 75 78

--Indicates congener was not 1 of 4 most abundant for that sample.

The water samples were characterized by the abun-
dance of three congeners: 2,2'-dichiorobipbenyl
(2,2'), 26-dichlorobiphenyl (26) and 26,2'-trichloro-
biphenyl (26,2') (Table 2). These three together
contributed from 51 to 78% of the total PCB
concentration. The next most abundant congener was
25,2'-trichlorobiphenyi (25,2') (Table 2).
The congener pattern of PCBs in C. tentans differed
substantially from that in the water (Fig. 1; Table 2).
Total PCB concentrations for the insects ranged from
3 3 n g g -~drywt in the control samples (mean of
111 ng g- ~ for three replicates) to 7540 ng g- ] after
96 h (mean of 6160 ng g-~ for three replicates).
The water pattern was dominated by 2 or 3 di- or
trichlorinated congeners, contributing approx. 50%
of the total concentration. The C. tentans samples
were characterized by a greater number of congeners
with the most abundant contributing only 5-10%
(Table 2). From 1 to 24 h, 2,2' and 26,2' were often
most abundant congeners, as in the water samples.
However, at 48, 72 and 96h, neither of these
compounds was dominant in the larvae. Congeners
taken up more slowly, such as 24,2'5'-tetrachloro-
biphenyl (24,2'5'), 236,4'-tetrachlorobiphenyl (236,4')
and other tri- and tetrachlorinated congeners,
were most abundant in 14 of 18 samples, although
each contributed less than 10% of total PCB
concentration.
Congener uptake--July
For each of the congeners of interest, and for total
PCBs, K~ and K2, concentration factors, and days to

(A) HUDSON RIVERWATER

(Sample 85-77, July exposure)
104 ng/I

(s)

~
Chironomu: tentans
urs)

i I I I ]
o 22.~ 5o.24 7e.ls ~o~.ss
ELUTION TIME (mln)
Fig. 1. Sample ehromatograms from Hudson river water (A) and Chironomus tentans (B) PCB analyses,
showing patterns of congener abundance.
324 M.A. NOVAKet al.

equilibrium are shown in Table 3. The congener
specific uptake and estimated exponential model are
illustrated in Fig. 2.
K~ values were large and positive; K2 values were
less than one in nearly all cases, indicating that
the ability of the test organisms to eliminate most
congeners was limited. K 2 values decreased with
increasing chlorination, consistent with the more
lipophilic nature of the highly chlorinated congeners.
Concentration factors (CF) ranged from approx.
4000 to 300,000 (Table 3).
September results
Reported 0 h total PCB concentrations in the test
organisms (mean 11 l0 ng g-J) were approximately an
order of magnitude higher than the mean in July;
2-chlorobiphenyl and 2,2'-dichlorobiphenyl were
particularly elevated. The source of this difference is
unknown, but contamination in the laboratory
colony itself was eliminated by analyzing insects and
water taken directly from the rearing tanks. Samples
of food and organic debris collected from the tanks
were also analyzed, but elevated PCB levels were not
detected. Several congeners: 24,2'; 234,4'; and
245,2'5', which separated satisfactorily in the July
analysis, were not present in the chromatograms from
the September exposure. Also, as seen in Table 1, the
September total PCB water concentration was nearly
constant over time at approx. 35 n g g - J , less than
every July value.
DISCUSSION
In the July exposure, the range of Kj and K 2 values
calculated for the selected congeners is consistent
with the values calculated by Muir et al. (1983) from
a laboratory study of 245,2'4'5'-hexachlorobiphenyl
uptake by Chironomus tentans. They reported that for
this congener, equilibrium was not reached after 96 h.
Similarly, some of the more highly chlorinated con-
geners in the present study did not attain equilibrium
during the 96 h exposure period.
With additional method development, K 1 and K2
uptake and elimination constants may be useful in
predicting PCB levels expected in a test organism,
such as C. tentans, from water column concen-
trations, and provide a model for predicting concen-
tration patterns in other river organisms. In this
study, the July replicate was successful in defining
the C. tentans/water PCB relationship, but the Sep-
tember replicate was unsuccessful. Detailed analysis
of the September exposure revealed that, unlike the
July results, two or even all three of the values do not
agree with equation (3), and an assumed normal
distribution for the residuals. Calculated K 2 values for
four congeners: 236,3'; 235,2',6'; 25,3'4'; and 24,3'4',
were negative. These negative values result from
an inability to identify sets of outlying points; the
data are not sufficiently self-consistent to identify
them from outliers. Between the two replicates, no
congener was found to have Kt and K 2 statistically
indistinguishable from July to September. These are
noted in Table 3.
Statistical tests were conducted (P <0.05) to
compare the estimated parameters between replicates.
For seven congeners, the Kj estimates were not
significantly different and for six other congeners, the
K 2 values were not significantly different.
Since contamination of the laboratory colony was
ruled out as a reason for the unexpected September
results, analytical error was suspected as the cause
of the discrepancies. The September results were
included here to point up potential problems with this
method of congener uptake measurements.

Table 3. Uptake and elimination constants, concentration factors (ng g- ~dry wt), and
estimated time to reach equilibrium (rounded to 3 significant figures) for Chironomus
tentans larvae in the Thompson Island pool of the Hudson river, July 1985
PCB KI CF K2 Days to 90%
congener ( u p t a k e / h ) ( K I / 1 ( 2 ) (elimination/h) equilibrium
2,2' 2370 5830 0.406 0.2
26* 1040 4190 0.247 0.4
26,2'* 6300 5950 1.060 0.1
2,4't 10,600 116,000 0.091 1.1
25,2'* 1860 37,000 0.050 t .9
24,2' 1930 56,700 0.034 2.8
26,4't I 120 47,800 0.023 4.1
246,2't 1730 42,800 0.041 2.4
25,3't 1920 156,000 0.012 7.8
25,4' 4390 258,000 0.017 5.6
24,4'* 5670 175,000 0.032 3.0
25,2'5' 3590 77,500 0.046 2.1
24,2'5'* 4670 117,000 0.040 2.4
24,2'4'* 4840 131,000 0.037 2.6
236,3' 2500 208,000 0.012 8.0
236,4't 5610 138,000 0.041 2.4
235,2'6' 5080 177,000 0.029 3.3
25,3'4' 8120 294,000 0.028 3.5
24,3'4'* 6660 362,000 0.018 5.2
234,4' 18,500 336,000 0.055 1.7
245,2'5' 2550 141,000 0.018 5.3
*, tCongeners for which Kt (*) or/(2 (t) values calculated for the September exposure
were not significantlydifferent from values calculated for July results.
 325
Determination of PCB rate constants

2.5 2 4.'

0.5 ~,a"~O . ~.3~,Or'

.,"o ,=2~ "
0 20 22 24 26 28 30 32 34. 36 38 4.0 ,t~',3~
x',-
RETENTIONTIME IN MINUTES

2.5

o.s " "l, =" ~ 4 ~.t,l"

o o ~d
4.8 4.8 5o ~2 ,~ ~6 s8 6o ,~o~
RETENTIONTIME IN MINUTES

c . . ~.~.y

2.5 24,2'5' , / / I ~ /

i~ ~ ., .,/, /;/./..x ,,~.o':,:,,

~~ .~ 7././:/.J~ X~> "
.~- at'.-, ~ ,,t.~ 0 ;.7" .,,/~o S "~
o : i oO ~ d' *"-
'~

RETENTION TIME IN MINUTES

Fig. 2. July congener specific uptake and estimated exponential model.

326 M.A. NOVAKet al.
The validity of the method depends on how closely
organism concentrations can be predicted from water
concentrations by use of equation (3). Muir et al.
(1983) found that first order kinetic rate equations as
used here were appropriate for describing organism
uptake. The exact solution to differential equation
(1):
CL( T) = KI .fCw(z) e -x2(r-~) dz
has been approximated by assuming Cw(t) is linear
between sampling times; the sampling intervals must
be sufficiently close together for the approximation to
be accurate. The integral in the above equation points
out a possible advantage in examining the organism
concentrations: they are the total response to the
input function Cw(t). The water concentration may
be highly irregular at times when PCB load is rapidly
changing, as during continued disposal, natural sedi-
ment scouring or during river dredging. Under these
conditions there is no assurance that sequential water
samples will capture transient concentration peaks
and troughs. Previous studies (Derr and Zabik, 1974;
Lynch and Johnson, 1982; Macek et al., 1979) have
shown that for several other organic compounds,
uptake in invertebrates is almost entirely from the
water column rather than from dietary sources.
Comparison of CL and Cw may therefore provide
verification that no significant peaks are being
missed. Once uptake constants have been fully
defined, use of Chironomus tentans to delineate water
concentrations of biologically important congeners
during the period of exposure will be another appro-
priate use for this method. Since these congeners are
preferentially accumulated by organisms, resulting in
high concentrations relative to water, the problems of
direct measurement of low or fluctuating water
column concentrations would be eliminated. This is
illustrated by comparison of results of the present
study with those of previous studies. The PCB
concentrations in the water samples differ from those
reported by Bush et al. 0985) for water samples
collected in the Thompson Island pool in 1983. They
reported substantially higher total concentrations
(approx. 500 ng l-1), and found 2-chlorobipbenyl (2)
to be one of the most abundant congeners at this
location. In the present study, 2-chlorobiphenyl was
not present above the limits of detection in any of the
water samples, although it was detectable in some C.
tentans samples.
Wood et al. (1987) expressed caution about the use
of aquatic organisms for monitoring xenobiotic com-
pounds such as PCBs, since congener uptake varied
both among and within species. However, within one
model species, differences in congener uptake are
problematic only if total PCB concentrations are
measured, but are not a limitation if congener-specific
analysis is conducted. In selecting specific congeners
for use in predicting PCB uptake in river organisms,
congeners which yielded consistent results should be
used, in the absence of further testing. Several of the
congeners studied--24,2'; 23,2'; 23,2'3'; 234,4'; and
245,2'5'--were not consistently separated by the
Apiezon L column. Since the less lipophilic congeners
2,2'; 26; 26,2'; and 2,4' are more water soluble and
appear to be variable in the environment, they are
considered less desirable for use in this manner.
However, four congeners---26,4'; 246,2'; 25,4'; 24,4';
and 236,4'--had comparable Ks values in the July
and September replicates, and the differences in K]
values, while statistically significant, were small
enough to result in the estimated concentration
factors being within a factor of 4 (246,2') or less.
These congeners should be considered for further
predictive studies.
This method has potential usefulness in field appli-
cations requiring biological assessment tools. If some
contaminants are known to be preferentially concen-
trated, it provides a means of elucidating concentra-
tions to be expected in the natural biota from either
hypothesized or observed inputs [Cw(t)]. Placing test
organisms both within and outside a contaminated
area could be used to delineate concentrations found
in specific river organisms. Additionally, this method
could be used to measure initial concentrations in
organisms, and those obtained after clean-up proce-
dures are conducted.
Acknowledgements--This study was supported by a grant
from the United States Environmental Protection Agency
to the New York State Department of Environmental
Conservation's Hudson River PCB Reclamation Demon-
stration Project. The research was initiated by Dr Karl
Simpson, of the New York State Department of Health,
now deceased. We thank Lawrence Abele for field collection
and processing of the samples, and Robert Bode for assis-
tance throughout the project.
REFERENCES
Bush B. and Barnard E. (1982) Determination of nonpolar
chlorinated hydrocarbons and PCB in microsamples.
Analyt. Lett. 15(A20), 1643-1648.
Bush B., Connor S. and Snow J. (1983) High resolution gas
chromatographic analysis of nonpolar chlorinated hydro-
carbons in human milk. J. Ass. off. analyt. Chem. 66,
248-255.
Bush B., Simpson K. W., Shane L. and Koblintz R. (1985)
PCB congener analysis of water and eaddisfly larvae
(Insecta: Trichoptera) in the Upper Hudson River by
glass capillary chromatography. Bull envir, contain. Toxic.
34, 96-105.
Derr S. K. and Zabik M. J. (1974) Bioactive compounds in
the aquatic environment: studies on the mode of uptake of
DDE by the aquatic midge, Chironomus tentans (Diptera:
Chironomidae). Arch. envir, contain. Toxic. 2, 152-164.
Hartley D. M. and Johnston J. B. (1983) Use of the
freshwater clam, Corbicula manilensis as a monitor for
organoehlorine pesticides. Bull. envir, contain. Toxic. 31,
33--40.
Lynch T. R. and Johnson H. E. (1982) Availability of a
bexachlorobipbenyl isomer to benthic amphipods from
experimentally contaminated natural sediments. In
Aquatic Toxicology and Hazard Assessment (Edited by
Pearson J. G., Foster R. B. and Bishop W. E.),
pp. 273-287. American Society for Testing and Materials,
Philadelphia, Pa.
 Determination of PCB rate constants
Macek K. J., Petrocelli S. R. and Sleight B. H. III (1979)
Considerations in assessing the potential for, and signifi-
cance of, biomagnification of chemical residues in aquatic
food chains. In Aquatic Toxicology (Edited by Marking
L. L. and Kimerle R. A.), pp. 251-268. American Society
for Testing and Materials, Philadelphia, Pa.
Muir D. C. G., Townsend B. E. and Lockhart W. L. (1983)
Bioavailability of six organic chemicals to Chironomus
tentans larvae in sediment and water. Envir. Toxic. Chem.
2, 269-281.
Muir D. C. G., Townsend B. E. and Webster G. R. B. (1985)
Bioavailability of L4C-1,3,6,8-tetrachlorodibenzo-p-dioxin
and ~4C-octachlorodibenzo-p-dioxin to aquatic insects
in sediment and water. In Chlorinated Dioxins and
Dibenzt~furans in the Total Environment H (Edited by
W.R. 24/3---E
327
Kerth L. H., Rappe C. and Choudhary G.). Butterworth,
Boston, Mass.
Sanders H. O. and Chandler J. H. (1972) Biological magnifi-
cation of a polychlorinated biphenyl (Aroclor 1254) from
water by aquatic invertebrates. Bull. envir, contain. Toxic.
7, 257-263.
Townsend B. E., Lawrence S. G. and Flannagan J. F. (1981)
Chironomus tentans Fabricius. In Manual for the Culture
of Selected Freshwater Invertebrates (Edited by Lawrence
S. G.). Canadian Special Publication of Fisheries and
Aquatic Sciences No, 54. Department of Fisheries and
Oceans, Ottawa.
Wood L. W., Rhee G-Y., Bush B. and Barnard E. (1987)
Sediment desorption of PCB congeners and their bio-
uptake by dipteran larvae. Wat. Res. 21, 875-884.