the effects of kepone on the estuarine copepod acartia tonsa

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Biological Sciences Theses & Dissertations Biological Sciences

Summer 1982

The Effects of Kepone on the Estuarine CopepodAcartia tonsaJudith Marie WilsonOld Dominion University

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WILSON, JUDITH MARIETHE EFFECTS OF KEPONE ON THE ESTUARINE COPEPOD ACARTIA TONSA

OLD DOMINION UNIVERSITY M.S. 1982

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THE EFFECTS OF KEPONE ON

A.B

A Thesis Si

in P a r t i a l

THE ESTUARINE COPEPOD A c a r t i a t o n s a .

by

Jud i t h Mar ie Wi lson

. May 1977, Randolph Macon Women's Col l ege

j b m i t t e d to the Facu l t y of Old Dominion U n i v e r s i t y

F u l f i l l m e n t of the Requirements f o r the Degree of

MASTER OF SCIENCE

BIOLOGY

OLD DOMINION UNIVERSITY

August 1982

A p n r o v e a D y :

' D i r e c t o r )


Copyri ght by J ud i t h Mar ie Wi lson 1982

A l l Rights Reserved


ABSTRACT

The E f f e c t s o f Kepone on the Es t u a r i n e Copepod A c a r t i a tonsa

J u d i t h Mar ie Wi lson Old Dominion U n i v e r s i t y , 1982

D i r e c t o r : Raymond W. Alden

Due to the c on tami na t ion of a 113 km reach of t he James

R i v e r , Kepone poses a ser i ous env i ronmenta l t h r e a t to the

Chesapeake Bay. The pupose of t h i s study is to determine

the a c u t e l y t o x i c and s u b le t h a l l e v e l s of Kepone f o r the

e s t u a r i n e copepod A c a r t i a t o n s a , and to i n v e s t i g a t e what

e f f e c t s those l e v e l s may have on f i l t r a t i o n r a t e s .

Kepone was determined to be a c u t e l y t o x i c to t o n s a ,

w i th a 96 hour LC50 o f 4 . 9 6 u g / 1 . D u n a l i e l l a t e r t i o l e c t a was

s e l e c t e d as the food source f o r the g r az ing e x p e r i me nt s .

Kepone concent r a t i o n s of 0 . 046 ug/1 s i g n i f i c a n t l y ( 0 . 0 5

l e v e l ) reduced the f i l t e r i n g r a t e under c o n d i t i o n s of

chron ic exposure. The l e s s e r c on c e n t r a t i o n s of 0 . 025 and

0 . 0046 ug/1 of kepone were not s i g n i f i c a n t l y d i f f e r e n t from

the c o n t r o l s . I t is hoped t h a t research i n t o the chron ic

s t ress of t h i s p e s t i c i d e may p o t e n t i a l l y prov i de i n f o rm a t i o n

which would be useful in the o v e r a l l unders tand ing of the

s ub le t ha l impact of c h l o r i n a t e d p e s t i c i d e s on a global

s e a l e .


This work is ded ica ted to my f a t h e r ,

James E. Wi lson J r .

and my l a t e mother Ethel S. Wilson


This acknowledges the spe c i a l help of Beth Heste r in

c u l t u r i n g D u n a l i e l l a t e r t i o l e c t a . Brook Haven Na t i ona l

L a b o r a t o r i e s of Upton, New York provided t h e P u n a ! i e l l a

t e r t i o l e c t a c u l t u r e . The s u b - c u l t u r e was obta ined from the

department of oceanography at Old Dominion U n i v e r s i t y . I

would a lso l i k e t o thank J e f f Hal l f o r the a ss is t anc e in gas

chromotography; and Dr . Raymond W. Alden f o r the b e n e f i t of

his knowledge and unending a s s i s t a n c e .


Table of Contents

LIST OF TABLES............................................................................v i i

LIST OF FIGURES........................................................................v i i i

Chapter

1. INTRODUCTION....................................................................1

2. MATERIALS AND METHODS.............................................. 10

3. STATISTICAL ANALYSES................................................. 15

4. RESULTS............................................................................16

5. DISCUSSION......................................................................18

BIBILIOGRAPHY................................................................................22

APPENDIXES

A. Tables..............................................................................26

B. Figures............................................................................39


vi i

LIST OF TABLES

TABLE Page

1. List of Registered Producers of Kepone Products....................................26

2. Average Environmental Kepone Concentrations............................................27P

3. Kepone in Chesapeake Bay Fish.........................................................................28

4. Kepone in James River Fish............................................................................... 29

5. Calculation of Kepone Concentrations Recovered by EC-GC...................30

6. Calculations for F ilte rin g and Feeding Rates..........................................31

7. Results of EC-GC Kepone Analysis from Collection S ite ....................... 32

8. Acartia tonsa Acute Toxicity Bioassay M ultiple Range Test-Duncan Procedure for the 0.05 le v e l........................................................... 33

9. Puna!iel la te rtio le c ta Chronic Toxicity Test. M ultipleRange Test-Duncan Procedure for the 0.05 le v e l.....................................34

10. Acartia tonsa Grazing Experiment Under Chronic Exposure.M ultip le Range Test-Duncan Procedure for the 0.05 le v e l..................35

11. Data from Carbon Analysis of Dunaliella te rtio le c taCulture Averaged from Three R eplicates....................................................36

12. Derived F ilte r in g and Feeding Rates of Acartia tonsaon Dunaliella te r t io le c ta ................................................................................37

13. Computer LC,q and LC5q Values with 95% Confidence Limitsfor 96-Hour Exposure Acartia Tonsa............................................................. 38


LIST OF FIGURES

FIGURE PAGE

1. Similar Molecular Structures of Kepone and Mirex.................................39

2. Hopewell, V irg in ia and Bailey Bay James River .....................................40

3. James River Sediment Concentrations from Hopewell, V irg in iato Newport News, V irg in ia , August 1976 .................................................. 41

4. Collection Site at Long Creek Bridge......................................................... 42


1

I n t r o d u c t i on

Kepone ( d e c a c h l o r o o c t a h y d r o - 1 , 3 , 4 , - m e t h a n o - 2 H - c y c l o b u t a

( c d ) - p e n t a l e n - 2 - o n e ) i s a c h l o r i n a t e d organ ic compound

s i m i l a r t o such p e s t i c i d e s as c h l o r dan e , DDT, a l d r i n ,

d i e l d r i n , and e n d r i n . Many of the physica l and chemical

p r o p e r t i e s of Kepone which s a t i s f i e d i t s i n d u s t r i a l use are

the same p r o p e r t i e s which make Kepone a se r ious t h r e a t to

the env i ronment . Kepone i s a whi te to tan c r y s t a l l i n e s o l i d

which subl imes w i th decomposi t ion a t 350° C. I t s vapor

pressure is very low and s o l u b i l i t y in water is only 0.4% at

100°C (Dawsey, 1 9 6 4 ) . Kepone is known to be s t r o n g l y

l i p o p h i l i c and b i o a c c u m u l a t i v e , w i t h a h a l f l i f e of one

decade or more ( S u t a , • 1 9 7 8 ) . Chemical s t a b i l i t y in n a t u r e ,

an a f f i n i t y f o r the b i o t a , and a broad spectrum of

b i o l o g i c a l e f f e c t s causes Kepone to be an env i ronmenta l

hazard of e c o l o g i c a l concern.

Kepone has been used to e r a d i c a t e roaches, a n t s , rust

m i t e , p i neapp le mealy bug, potato w i r e worm, and tobacco

w ir e worm ( S u t a , 1 9 7 8 ) . Kepone is a cumul at ive poison,

meaning t h a t t o x i c l e v e l s are reached a f t e r smal l amounts

are taken over a long per i od of t i m e . The p e s t i c i d e a f f e c t s

i ns ec t s as a nerve poison, i n t e r f e r i n g wi t h the passage of

impulses of the nervous system. I n i t i a l t o x i c a c t i on may be


2

a r educ t i on of ATP syn the s i s by i n h i b i t i o n of o x i d a t i v e

p ho sphor y l a t ion in the m i t o c h o n d r i a , which has been

i n d i c a t e d by the i n h i b i t i o n of the enzymat ic f u n c t i n of

ATPase ( S t e r r e t t , 1977 and Desa i ah , 1 9 7 5 ) . This e f f e c t is

p a r t i c u l a r l y i m p o r t an t to mar ine organisms whose p o l a r

l i p i d s on e x t e r n a l membranes (based on Na+ + K+-dependent

ATPases) are engaged in i o n i c r e g u l a t i o n ( S a r g e n t , 1 9 7 6 ) .

Kepone was f i r s t r e g i s t e r e d as a p e s t i c i d e wi th the

USDA in 1959, w i t h the o r i g i n a l pa t en t s granted in 1952 . I t

i s found both as a degre da t ion product and a contaminant of

M i r e x , another c h l o r i n a t e d p e s t i c i d e s i m i l a r in chemical

c h a r a c t e r i s t i c s and b i o l o g i c a l a c t i v i t y (Mackenthun, 1978;

S t e r r e t t , 1 9 7 5 ) . Mi rex s lowly biodegrades t o Kepone in the

presence of anaer ob i c microbes and l i g h t ( n a t u r a l or

a r t i f i c i a l ) . Due to the presence of an oxygen atom in the

m o l ecu l a r s t r u c t u r e , Kepone is more water s o l u b l e ( through

hydrogen bonding) than Mi rex ( F ig u r e 1 ) . These cyc lo d ie n e

(or c y c l o d i o l e f i n e , a c y c l i c hydrocarbon which con ta i ns two

double bonds of the general formula C C^H^) p e s t i c i d e s are

s yn thes i zed from he x ac h l o ro cy c lo p en t ad i en e (HEX) .

The product i on dates of Kepone span from 1958 to 1975,

the main manufacturers being A l l i e d Chemical Company wi th

p l a n t s at B a l t i m o r e , Maryland and Hopewel l , V i r g i n i a ( 19 66 -

1 97 3 ) ; L i f e Science Products Company at Hopewel l , V i r g i n i a

( 1 9 7 3 - 1 9 7 5 ) ; and Hooker Chemicals and P l a s t i c s Corp or a t i on

at Niagara F a l l s , New York. See Table 1 f o r a d d i t i o n a l

producers of Kepone p r odu ct s . E v e n t u a l l y , more than 70 out


3

of 150 employees at t he L i f e Science Products Company in

Hopewell were diagnosed as having Kepone po i son i ng .

Symptoms ranged from s l u r r e d speech and loss of memory to

l i v e r damage and s t e r i l i t y (Mackenthun, 1 9 7 8 ) .

The L i f e Science p l an t manufactured between 1360 t o

2720 kg of Kepone per day, o p e r a t i n g nonstop seven days a

week. Between March 1974 and A p r i l 1975, moni tored

suspended a tmospher ic p a r t i c u l a t e s conta i ned at l e a s t 40%

Kepone ( S t e r r e t t and Boss, 1 9 7 7 ) . Kepone e f f l u e n t s from the

L i f e Science p l a n t went d i r e c t l y to the Hopewell sewage-

t r e a t m e n t p l a n t , i n h i b i t i n g normal b a c t e r i a l a c t i o n , causing

a shutdown of sewage- t rea tment o p e r a t i o n s . Unt rea t ed sewage

then p o l l u t e d t h e James Ri ver and ass oc i a t e d w a t e r s . As a

r e s u l t A l l i e d Chemical was f i n e d 13 . 2 m i l l i o n d o l l a r s , the

l a r g e s t p o l l u t i o n p e n a l ty a ga ins t any U.S. company, and L i f e

Science Products was f i n e d 3 . 8 m i l l i o n d o l l a r s f o r t h e i r

p a r t i c i p a t i o n in p o l l u t i n g t h e James Ri ver w i th Kepone

( S t e r r e t t and Boss, 1977) ( F i g ur e 2 ) .

U .S . pr oduct i on in 1974 alone r e s u l t e d in app ro x i mat e l y

3 63 . 00 0 kg of Kepone, 90% of which was expor ted to La t in

America, A f r i c a , and As i a , making Kepone a wor ld wide

concern . The amount of Kepone l o s t from the L i f e Science

op e ra t io n was app ro x i mat e l y 91 , 000 kg out of an es t imated

777 .000 kg produced ( Su t a , 1 97 8 ) . Lost Kepone was the

combined r e s u l t of atmospher ic r e l e a s e , waste water

d i s c h a r g e , equipment m a l f u n c t i o n s , r e l e as e of substandard

product i on batches , and car e l es s dumping of waste loads at


4

t he Hopewell l a n d f i l l . The present s t a t us of Kepone is t h a t

i t i s no l onger manufactured. However, as of October 1976,

t h e r e were 227 kg o f Kepone l e f t in s t o c k p i l e s in the

c o n t i n e n t a l U . S. which were s u f f i c i e n t t o cont i nue meet ing

the demands as a component of ant and roach b a i t and t ra p s

f o r at l e a s t t h r ee years ( Su t a , 1 97 8 ) .

In September of 1975 the EPA informed the FDA of James

R i v e r con tami na t ion and in February of 1976 the EPA

recommended a c t i o n l e v e l s to the FDA f o r s e i z u r e of

co n ta i m i n a t e d f i n f i s h and s h e l l f i s h . These l e v e l s begin at

0 . 3 ug/g f o r f i n f i s h , o y s t e r s , and clams, and 0 . 4 ug/g f o r

crabs ( S u t a , 1 97 8 ) . Schimmel, et a l . ( 1979 ) r epor t ed t h a t

based on t h e i r data "Kepone may be a f a c t o r in the d e c l i n e

of the James Ri ver crab f i s h e r y . " Publ ished average

env i ronmenta l c on c en t r a t i o n s of Kepone in both James River

seafood, and Chesapeake Bay seafood may be found in Tables

2, 3 and 4 of the Appendix (Mackenthun, 1 9 7 8 ) . Today Kepone

l e v e l s remain under c lose s c r u t i n y . Kepone has now been

found in land animals which are ass oc i a te d w i th mar ine

animals v ia the food web. The EPA es t ima te s 45 , 300 kg of

Kepone is in the James R i v e r , which is r e spo ns i b l e f o r 15%

a n n u a l l y of the f re s h water f low i ng i n t o the Chesapeake Bay,

and 10% a n n u a l l y of the sediments expected to move i n t o the

Bay ( S u t a , 19 7 8 ) .

Due to contaminat ion of a 113 km reach of the James

Ri ver ( t h r e a t e n i n g the Chesapeake Bay) ( F i g u r e 3)

p r e l i m i n a r y t rends in Kepone a n a ly s i s were concerned wi th


5

the chemical aspects of t r a n s p o r t , downstream

t r a n s f o r m a t i o n , and exposure to mar ine l i f e . The p o t e n t i a l

human h e a l t h hazard from e d i b l e t i s s u e s t i m u l a t e d the t rend

in t o x i c i t y , up t ake , and depuraion s t ud i es of the p e s t i c i d e

in marine mo l lus ks , c r us t ac eans , and f i n f i s h . Present

research i d e n t i f i e s the need f o r a more comprehensive

knowledge in assessment of the b i o l o g i c a l and e c o l o g i c a l

impact of Kepone on the mar ine env i r onment . In l a b o r a t o r y

e v a l u a t i o n s of env i ronmenta l impact , the chron ic t o x i c i t y

and b i o c o n c e n t r a t i o n p o t e n t i a l of Kepone are as i mpor tant as

i t s acute t o x i c i t y . Research i n t o the chron ic s t re ss of

t h i s p e s t i c i d e i s of e co l o g i c a l s i g n i f i c a n c e on a l oca l

l e v e l and in the Chesapeake Bay, and may p o t e n t i a l l y prov ide

i n f o r m a t i o n which would be usefu l in the o v e r a l l

understanding of the sub l e t ha l impact of c h l o r i n a t e d

p e s t i c i d e s on a g lobal s c a l e . Kepone con tami na t ion is a

c u r r e n t problem w i t h f u t u r e economic i m p l i c a t i o n s in regard

to the James R i ve r and Chesapeake Bay f i s h e r i e s and

e c o l o g i c a l l y in regard to the o t her b i o t a in the Bay.

The purpose of t h i s study is to de termine a c u t e l y t o x i c

and s u b le t h a l l e v e l s of Kepone f o r the e s t u a r i n e copepod

A c a r t i a tonsa and to i n v e s t i g a t e what e f f e c t s those l e v e l s

may have on f i l t r a t i o n and i n g e s t i o n r a t e s . There are

seveal reasons f o r using a copepod as a t e s t organism.

Zooplankton , because of t h e i r high metabol ism and rap i d

r e p r o d u c t i v e a b i l i t y , respond q u i c k l y to env i ronmenta l

changes. Th e re f or e zooplankton p op u l a t i on dynamics may


6

prove to be s e n s i t i v e i n d i c a t o r s of the q u a l i t y of t h e i r

s ur ro un d in g s . Copepods are regarded as g e n e r a l l y

r e p r e s e n t i n g the dominant group among e s t u a r i n e and mar ine

zooplankton ( P a r r i s h and Wi l son , 1978; Bougis, 1976; and

Deevey, 1 9 5 6 ) . A c a r t i a tonsa is g e n e r a l l y the dominant

z o o p la nk t or along most of the eas te rn seaboard. Copepods

are a p a r t i c u l a r l y i mp or tan t food source t o m i g r a t i n g

species t h a t use e s t u a r i e s f o r breed ing grounds and

n u r s e r i e s . A c a r t i a tonsa is of spe c i a l e c o l o g i c a l

s i g n i f i c a n c e at the f i r s t consumer l e v e l in f i s h e r y cha ins ,

p l a y i n g a v i t a l r o l e in the energy t r a n s f e r of h i gher

t r o p h i c l e v e l s ( P a r r i s h and Wi l son , 1 9 7 8 ) . Because t h i s

Copepod occurs in no t eab le q u a n t i t i e s in l i t t o r a l zones, i t

plays a major pa r t in the n u t r i t i o n of young and

pianktophagous f i s h . Research has a lso shown t h a t Kepone

can be taken up and bioaccumulated by zooplankton (Bahner ,

et al . , 1 977 ) .

A c a r t i a tonsa may be among the most s e n s i t i v e of

e s t u a r i n e organisms, about two orders of magni tude lower in

t o l e r a n c e t o t o x i n s than grass shrimp (Palaemonetes p u g i o )

(APHA, 1 9 7 5 ) . There is a vast amount of i n f o r m a t i o n

pub l i shed on the phys io logy and b i o l ogy of _A. tonsa

(Conover , 1956; Durbin and Durb in , 1978; Gonzales, 1974;

J e f f r i e s , 1962; K h a t t a t , 1976; Lance, 1965; Landry, 1976;

M i l l e r , et a l . , P a r r i s h and Wi l son , 1975; Reeve and W a l t e r ,

1977; S e k i g u c h i , et a l . , 1980; and Wyman and O'Connors, 1980

among o t h e r s ) . Standard Methods (APHA, 1975) suggests t h a t


7

the f o l l o w i n g are pr ime c o n s i d e r a t i o n s in choosing a t e s t

organism: a) s e n s i t i v i t y to the t e s t m a t e r i a l ; b)

geographica l d i s t r i b u t i o n , abundance, and a v a i l a b i l i t y ; c)

r e c r e a t i o n a l , economica l , and e co l o g i c a l importance l o c a l l y

and n a t i o n a l l y ; d) a v a i l a b i l i t y of c u l t u r e methods and

knowledge of env i ronmenta l r equ i rement s; e) general physica l

c o n d i t i o n and f reedom from p a r a s i t e s and d i s e a s e ; and

f i n a l l y f ) s u i t a b i l i t y f o r bioassay t e s t s . I n f o rm a t i on thus

f a r prov ided i n d i c a t e s h_. tonsa may be a we l l s u i t ed t e s t

organism by meet ing t he prime c on s i d e r t i o n s l i s t e d above.

In copepods t h e r e are two p r i n c i p a l ways of

n u t r i t i o n : f i l t r a t i o n and p r e d a t i o n . A c a r t i a tonsa is a

f i l t e r f e e d e r . This could be i mpor tan t because accumulat ion

of Kepone on p l a n t d e t r i t u s may be an i mpor tan t mechanism by

which i t e n t e r s e s t u a r i n e food webs ( D r i f m ey e r et a l . ,

1 9 8 0 ) . Because of Kepone's l a r g e mol ecu l ar weight and i t s

low s o l u b i l i t y in w at e r i t has a tendency to s e t t l e in

a q u a t i c bodies and accumulate in the sediments ( S t e r r e t t and

Boss, 1 9 7 7 ) .

Kepone l e v e l s in the water column are not b e l i e v e d to

be c on c e n t r a t e d enough to r e s u l t in acute t o x i c i t y a l though

t h e r e is no proof f o r t h i s . However chron ic exposure to low

l e v e l s could produce s ub l e t h a l e f f e c t s which are not r e a d i l y

a p p a r e n t , y e t d i s r u p t i v e to the system over a per i od of

t i me . The l i t e r a t u r e c a l l s f o r a b e t t e r unders tand ing of

the more s u b t l e e f f e c t s r e qu i r e d to assess envi ronmental

hazards of Kepone and i t s f u t u r e impact (Hansen, et a l . ,


8

1976; Hansen, e t a l . , 1977a; Schimmel and Wi lson, 1977;

Wyman and O'Connors, 1 98 0 ) .

Hansen, Goodman, and Wilson (1977a) upon t e s t i n g the

chron ic e f f e c t s o f Kepone on Sheepshead Minnows r epor t ed

t h a t :

Growth, r e p r o d u c t i o n , or s u r v i v a l was a f f e c t e d by a l l c o n c e n t r a t i o n s t e s t e d . I t is g e n e r a l l y accepted t h a t 0.01 of the acute t o x i c i t y of a p e r s i s t a n t o r gan ic chemical to an organism should be p r o t e c t i v e of a s pe c i es . Our data however i n d i c a t e t h a t a Kepone c o n c e n t r a t i o n o f 0.001 of t h e 96 hr LC50 f o r Sheepshead Minnows a f f e c t e d t h i s species d e t r i m e n t a l l y in chron ic t o x i c i t y t e s t s .

Another study of the chron ic e f f e c t s of Kepone on e s t u a r i n e

organisms r e p or t s t h a t s u r v i v a l , growth, and repr oduct i on of

mysid shrimp (Mysidopsis b a h i a ) were decreased (Hansen, et

a l . , 1 9 7 7 b ) . Wyman and O'Connors (1980 ) i n d i c a t e t h a t

perhaps the i n cr e a se of PCB ( s y n t h e t i c c h l o r i n a t e d

hydrocarbons s i m i l a r t o DDT and Kepone) accumulat ion due t o

i n g e s t i o n of contaminated phy top lank ton is s u f f i c i e n t to

i n c r ease m o r t a l i t y and reduce f e c u n d i t y among the

copepods. I t i s t h e r e f o r e reasonable to hypothes ize t h a t

s u b le t h a l q u a n t i t i e s of Kepone in chron ic exposure could

a f f e c t the physio logy of A c a r t i a t o n s a . Whether or not the

copepod feeds normal ly under c o n d i t i o n s of chron ic exposure

t o Kepone w i l l i n d i c a t e i f the copepod has been

p h y s i o l o g i c a l l y a f f e c t e d . The l e v e l and q u a l i t y of energy

i nput ( f rom inges t ed food) i s v i t a l to an organism's

f u n c t i o n i n g and s e l f p r e s e r v a t i o n . A c a r t i a t o n s a , having a

high metabol ism and a rap i d t u r n o v e r , may have to s a c r i f i c e


9

energy f o r metabol ism and growth, c u t t i n g down on a v a i l a b l e

energy d i r e c t e d towards r e p r o d u c t i o n . Studies by P a r r i s h

and Wilson (1978) i n d i c a t e t h a t tonsa females have

reduced energy reserves in comparison wi th some "deep-water

or near su r face c o l d - w a t e r copepods" which use wax e s t e r s as

energy reserves in t imes of food d e p r e i v a t i o n . They

cont inue to observe t h a t the lack of f ood- energy a p p a r en t l y

r e s u i t s in a lower egg p r o du c t i o n . The i n g e s t i o n ra tes and

growth e f f i c i e n c y of organisms on an i n d i v i d u a l basis may

t her eby a f f e c t the success and s u r v i v a l of the popu l a t ion as

a whole .


10

M a t e r i a l s and Methods

Exper imenta l animals were c o l l e c t e d weekly between June

1981 and October 1981. The c o l l e c t i o n s i t e was Long Creek

Br idge l o c a t e d j u s t east of Lynnhaven I n l e t in V i r g i n i a

Beach, V i r g i n i a ( F i g u r e 4 ) . Long Creek j o i n s Lynnhaven Bay

t o Broad Bay on the west w i t h r e s i d e n t i a l and business areas

to the nor th and south . During the c o l l e c t i o n p er i od

s a l i n i t i e s f l u c t u a t e d from 24.1 to 2 4 . 9 ° / o o and tempera tures

ranged from 1 6 . 3 to 2 0 . 1 ° C . The average d i s so l v ed oxygen

l e v e l was 8 . 4 mg/1. Heavy t i d a l f low both ebb and f lood

made t h i s l o c a t i o n i de a l f o r p lankton tows.

For c o n s i s t a n c y , a l l tows were taken on incoming

t i d e s . A 250 micron mesh, 0 . 5 meter p lankton n e t , o u t f i t t e d

w i t h a PVC cod end b o t t l e , was u t i l i z e d f o r the

c o l l e c t i o n s . By l ower i ng the net d i r e c t l y below Long Creek

B r i d g e , a good h o r i z o n t a l tow could be mai nta ined by t i d a l

f l o w . A f t e r severa l p r e l i m i n a r y tows had been made, i t was

decided t h a t due to high j e l l y f i s h c o n c e nt r a t i on s in t h i s

a r e a , a t h r e e minute tow per iod was a p p r o p r i a t e . On each

c o l l e c t i o n t r i p , ten tows were t a k e n . A p o r t a b l e

i n d u c t i o n s a l i n o m e t e r was used to measure c o n d u c t i v i t y ,

s a l i n i t y , and t e m p e r a t u r e , BOD b o t t l e s were a lso f i l l e d f o r


11

l a b o r a t o r y d e t e r m i n t i o n of D.O. by W in k le r T i t r a t i o n in

c o n j u n c t i o n wi th the use of a f i e l d D.O. m e t e r .

Water from the c o l l e c t i o n s i t e was t e s t e d f o r the

presence of Kepone. Samples were e x t r a c t e d as o u t l i n e d by

the EPA H ea l th E f f e c t s Research L a b o r a t o r y , Research

T r i a n g l e P a r k , N.C. ( W a t t s , e t a l . , 1 9 8 0 ) . Kepone

c o n c e n t r a t i o n s were determined by i n j e c t i o n of e x t r a c t s i n t o

a Shimadzu Model 6AM gas chromatograph w i t h an e l e c t r o n

cap tur e d e t e c t o r . The method of c a l c u l a t i o n of Kepone

c o n c e n t r a t i o n appears in Tab le 5 . A n a l y t i c a l p e s t i c i d e

standards (Kepone and M i r e x ) were ob t a i ned from the

r e f e r e n c e s t andards r e p o s i t o r y , ETD, HERL, EPA, Research

T r i a n g l e Pa rk , N.C.

The zoop lank ton samples were kept in an i n s u l a t e d

a er a t e d c o n t a i n e r u n t i l reaching t he l a b o r a t o r y , which was

always w i t h i n one hour of c o l l e c t i o n . To min imi ze any shock

the samples were a c c l i m a t e d to l a b o r a t o r y t e mper a t ur e ( 2 2 °

C) f o r a 24 hour p e r i o d , and then a d j us t ed to 2 5 ° / o o

s a l i n i t y . Adul t A c a r t i a tonsa were then sor ted out using a

Nikon Model 102 d i s s e c t i n g microscope and a wide bore

pi p e t t e .

Acute t o x i c i t y was determined f o r 96 hours in s t a t i c

b i oa ss a ys . The 96 hour LC10, LC50, and LC100 va lues were

based on nominal c o n c e n t r a t i o n s . The values w i th conf idence

l i m i t s were computed based on t he L i t c h f i e l d and Wilcoxon

( 1949 ) method of s t a t i s t i c a l a n a l y s i s . Checks were made at

12 hour i n t e r v a l s . C r y s t a l l i z i n g dishes (170 ml ) were


12

f i l l e d wi t h 150 ml each oxygenated a r t i f i c i a l seawater

( Standard Methods APHA, 1 97 5 ) , t r e a t e d wi t h t e c h n i c a l grade

Kepone d i s s o l v e d in 0.1 ml ace tone . This l e v e l of acetone

produced no s i g n i f i c a n t e f f e c t s on _A. t o n s a . A f t e r 45

minutes , ten a d u l t s were randomly p laced in each one of the

c r y s t a l l i z i n g d i s h e s , which were subsequent ly covered.

Feeding was omi t t ed f o r the d u ra t i on of the b i oass ays . Each

bioassay was performed in t r i p l i c a t e w i th both an acetone

and a clean water c o n t r o l . The Kepone f o r a l l stock

s o l u t i o n s was weighed out on a Cahn E l e c t r o n i c Ba lance.

Si ng l e a d d i t i o n s o f Kepone were admi n i s t e r ed to achieve the

t e s t l e v e l s t hroughout the e xp er iment s . There were two

reasons f o r t h i s d e c i s i o n . Zooplankton are f r e e f l o a t i n g

p op ul a t i ons in the water column t h a t move away from the

source of i nput along wi th the w a t e r . T h e r e f o r e they are

exposed to c o n t i n u a l l y decreas ing l e v e l s of p o l l u t a n t s .

Also s i n g l e a d d i t i o n s more c l o s e l y approximate the normal

ocean s i t u a t i o n i f the source of the p o l l u t a n t is from

o u t f a l l s , r i v e r s or dumping (Menzel and Case, 1 9 7 7 ) , which

i s the case w i th Kepone contami nat ion in the Chesapeake Bay.

The r e s u l t s of the acute t o x i c i t y t e s t s were used to

detemine which s u b le t h a l l e v e l s should be used in subsequent

t e s t i n g f o r chr on ic s t r e s s . Three d i f f e r e n t s u b l e t h a l

l e v e l s ( 0 . 0 0 4 6 , 0 . 0 2 5 , and 0 . 046 ug/1 ) of Kepone were used

to compare t h e i r r e l a t i v e e f f e c t s in a graz i ng exper iment

under c o n d i t i o n s of chron ic exposure . Chronic exposure in

t h i s case is de f i ned as 48 hours of prev ious exposure to


13

sub l e t ha l l e v e l s before t e s t i n g begins f o r 24 hours.

P r e p a r a t i o n of water and a d d i t i o n of _A. tonsa were c a r r i e d

out as in the acute t o x i c i t y b ioassays.

D u n a l i e l l a t e r t i o l e c t a was s e l e c t e d as the food source

because i t i s u n i c e l l u l a r , t h e r e f o r e p a r t i c l e m o d i f i c a t i o n

by _A. tonsa (as wi th chain forms) would not be a f a c t o r in

v i sual counts . D u n a l i e l l a t e r t i o l e c t a l i k e _A. tonsa i s

e u r y h a l i n e . Because _D. T e r t i o l e c t a is f l a g e l l a t e d ,

suspension throughout the water column f o r a 24 hour per iod

i s p os s ib l e (and p r e f e r a b l e ) in graz ing exp er iment s .

D u n a l i e l l a t e r t i o l e c t a was c u l t u r e d in ,lf / 2 " medium at a pH

of 8 . 0 , 22°C, and 25 ° / oo s a l i n i t y according to the methods

of G u i l l a r d (1 9 7 5 ) . Before using D . t e r t i o l e c t a in the

graz i ng exp er iment s , c u l t u r e s were exposed t o 0 . 0046 and

0 .046 ug/1 of Kepone f o r 72 hours each to determine any

r e l a t i v e d i f f e r e n c e s in growth r a te from the cont ro l

c u l t u r e .

A f t e r a 36 hour exposure p e r i o d , A_. tonsa a du l t s were

t r a n s f e r r e d to one l i t e r of f r e s h Kepone t r e a t e d water f o r a

12 hour s t a r v a t i o n p e r i o d , at which t ime p r e v i o u s l y exposed

D_. t e r t i o l e c t a c u l t u r e was added. The graz i ng exper iments

were c a r r i e d out f o r 24 hours in t r i p l i c a t e using both a

clean water and an acetone c o n t r o l . F i l t e r i n g and f eed i ng

r at es were de r ived from the c a l c u l a t e d graz i ng rates

according t o Edmondson and Winberg (1971) ( Tab l e 6 ) .

D u n a l i e l l a t e r t i o l e c t a was a lso ana lyzed f o r carbon

content using an Oceanogrphy I n t e r n a t i n a l Tot a l Carbon


14

Ana lyzer t o supplement the v i sua l counts using a P e t r o f f -

Hausser B a c t e r i a Counter . Ampules c o n t a i n i n g ^ . t e r t i o l e c t a

c u l t u r e were purged wi t h oxygen to remove any carbonate ions

and then were sea led w i t h o u t contami nat ion by the ampule

purging and s e a l i n g u n i t . The ampule a n a l y z i n g u n i t

( c o n t a i n i n g an Hor iba Model P I R-2000 General Purpose

i n f r a r e d a n a l y z e r ) was used to determine the c on c e n t r a t i o n

of carbon d i o x i d e r e s u l t i n g from the wet o x i d a t i o n of

or gan ic m a t t e r in the t e r t i o l e c t a c u l t u r e . F luoresence

a n a l y s i s was also a t tempted but the r e s u l t s proved to be too

i n c o n s i s t a n t f o r t he d e t e r m in a t i o n of f ee d i n g r a t e s .


15

S t a t i s t i c a l Analyses

Three parameters were s t a t i s t i c a l l y ana lyzed in order

to get a c l e a r e r p i c t u r e of the e f f e c t s of Kepone on A c a r t i a

t o n s a . These were 1) m o r t a l i t y of _A. tonsa due to acute

t o x i c i t y , 2) r e l a t i v e growth ra t es of D u n a l i e l l a t e r t i o l e c t a

exposed to Kepone, and 3) f i l t e r i n g r at es of _A. tonsa at

d i f f e r e n t s u b l e t h a l l e v e l s o f Kepone.

M o r t a l i t y data was t r e a t e d w i t h a l i n e a r r e g r e s s i o n , a

one-way a n a l y s i s of v ar i ance (ANOVA), and a Duncan's

m u l t i p l e range t e s t (Sokal and R o h l f , 1 9 6 9 ) , f o l l o w i n g the

t r a n s f o r m a t i o n of Kepone c o n c e n t r a i t o n to l o g s , and percent

m o r t a l i t y to a r c s i n e - fP . D u n a l i e l l a t e r t i o l e c t a growth ra t es

were t e s t e d f o r homogeniety using the one-way a n a l y s i s of

v a r i a n c e and a Duncan's m u l t i p l e range t e s t (Sokal and

R o h l f , 1 9 6 9 ) . F i l t e r i n g r a t e s were a l so t e s t e d by ANOVA in

a d d i t i o n to the Duncan's and the more s e n s i t i v e W i l l i a m ' s

t e s t ( W i l l i a m s , 1 9 7 1 ) .


16

Resul ts

The Kepone d e t e r m i n a t i o n procedure y i e l d e d 91.25%

recovery of t h e Kepone s tandard and 103% recovery of the

Mi rex s p i k e . R e p l i c a t e s of water samples from the

c o l l e c t i o n s i t e showed Kepone l e v e l s to be non de t ec t ab l e

( < 0 . 0 2 ug/1 ) (Table 7 ) . T h e r e f o r e , the copepods were

presumed to be from a c l ean c o l l e c t i o n s i t e w i t h no

s i g n i f i c a n t Kepone c o n t a m i n a t i o n .

The 96 hour LC50 f o r A c a r t i a tonsa was determined to be

4 . 9 6 ug/1 w i t h 9 . 4 ug/1 at the upper l i m i t and 2 . 6 ug/1 at

the lower l i m i t , as determined by the L i t c h f i e l d and

Wilcoxon Method ( 1 9 4 9 ) . The l i n e a r r egr ess ion between log

c o n c e n t r a t i o n and a r c s i n e m o r t a l i t y was shown to be p o s i t i v e

and h i g h l y s i g n i f i c a n t at a p of 0 . 0 1 . Homogenei ty of

va r ia nc e was t e s t e d by the B a r t l e t t ' s and Cochran's t e s t .

These showed the data be be homogeneous. The ANOVA and

Duncan's i n d i c a t e d t h a t each Kepone c o n c e n t r a t i o n ( 0 . 0 5 ,

0 . 5 , 5 . 0 , 5 0 . 0 , and 500 .0 u g / 1 ) was s i g n i f i c a n t l y d i f f e r e n t

(p o f 0 . 0 5 ) in the l e v e l of m o r t a l i t y t h a t was caused ( Tab l e

8 ).The ANOVA and Duncan's t e s t i n d i c a t e d t h a t t h e r e were

no s i g n i f i c a n t d i f f e r e n c e s (p of 0 . 0 1 ) between mean c e l l

number of _D. t e r t i ol ec ta in the con t r o l or the su b le t ha l

l e v e l s of (based on nominal concentrations o f Kepone)


17

0 . 0046 ug/1 or 0 . 046 ug/1 a t 72 hours of exposure (Table

9 ) . T h e r e f o r e , counts of _D. t e r t i o l e c t a f o r the graz ing

exper iments could be considered r e l i a b l e and u n a f f e c t e d .

To moni tor r e l a t i v e e f f e c t s of s u b le t h a l s t ress in the

graz i ng exper iments 0 . 0 4 6 , 0 . 0 2 5 , and 0 . 0046 ug/1 o f Kepone

were used. The l e v e l of 0 . 0 4 6 ug/1 was shown to be

s i g n i f c a n t l y d i f f e r e n t (p o f 0 . 0 1 ) from 0 . 025 or 0 . 0046

u g / 1 . The more s e n s i t i v e Wi l l i ams t e s t was used to

determine i f 0 . 025 or 0 . 0046 ug/1 were s i g n i f i c a n t l y

d i f f e r e n t from the c o n t r o l . The r e s u l t s y i e l d e d were the

same as those from t he Duncan' t t e s t . The two lower

c o n c e n t r a t i o n s were grouped as being not s i g n i f i c a n t l y

d i f f e r e n t from the c o n t r o l s (See Table 1 0 ) .


18

Di scussi on

Kepone was found t o be a c u t e l y t o x i c to _A. tonsa

a d u l t s . The 96 hour LC50 o f 4 . 9 6 ug/1 as determined by t h i s

s t udy , is c o n s i s t a n t w i t h the b e l i e f t h a t the s e n s i t i v i t y of

_A. tonsa may be about two orders of magni tude lower than

Palaemonetes pugio whose LC50 as determined by Hansen et a l .

(1 977b) i s 120 u g / 1 . Hansen et al . (1 977b) found in acute

t o x i c i t y t e s t s of the z o o p la nk t or Mysidopsis b a h i a , a 96

hour LC50 of 10 ug/1 of Kepone. The animals become l e t h a r g i c

be f or e d ea t h . There was no c o l o r change in t he organism as

a r e s u l t of p o i son i ng . A c a r t i a tonsa e x h i b i t e d the same

l e t h a r g y bef or e dea th , r e t a i n i n g normal c o l o r appearances.

When a d u l t A_. tonsa were t e s t e d f o r acute t o x i c i t y of

the f o l l o w i n g p e s t i c i d e s they ranked h i ghes t to l owest ;

toxaphene , d i a z i n o n , a z o d r i n , and then methyl p a r a t h i o n .

Kepone appears to rank wi th diaznon in i t s acute t o x i c i t y to

A_. t o n s a . Azodr in was f our t imes as t o x i c as methyl

p a r a t h i o n , and d i az i non was 100 t imes more t o x i c than

azodr i n ( Tab l e 13) ( K h a t t a t and F a r l e y , 1 9 7 6 ) . Un l i ke o ther

c y c l o d i e n e p e s t i c i d e s such as Kepone, toxap. iene has no

e f f e c t on the nervous system, r a t h e r i t is d i s t r i b u t e d

t hroughout the hemolymph. Azodr in (an organophosphorous


19

compound) u n l i k e Kepone is water s o l u b l e , which may enhance

i t s uptake and metabol ism of chronic l e v e l s , r e s u l t i n g in

q u i c ke r t o x i c a c t i o n . This could lend e x p l a n a t i o n to i t s

g r e a t e r t o x i c i t y to _A. tonsa than Kepone. Methyl par a t h i on

was found to be more t o x i c to Palaemonetes v u l g a r i s (96 hour

LC50 of 3ppb) than Kepone was to Palaemonetes pugio (96 hour

LC50 of 120ppm). Methyl p ar a t h i on was 10 f o l d more t o x i c to

A* tonsa than Kepone was found to be from the present s tudy .

In g r az ing exper iments of chron ic t o x i c i t y , Kepone was

shown to be d e t r i m e n t a l . Sub le tha l l e v e l s began at 0.01 of

the acute t o x i c i t y , which according t o EPA c r i t e r a , should

be p r o t e c t i v e f o r a s p e c i e s . The 0 . 046 ug/1 dose l e v e l

produced s i g n i f c a n t l y reduced f i l t e r i n g r a t e s . The 0.001

l e v e l of 0 . 0046 ug/1 y i e l d e d no d i f f e r e n t r e s u l t s than did

the c o n t r o l s or t he 0 . 025 ug/1 dose. In chron ic t e s t i n g of

ot her organisms Hansen, Goodman, and Wi lson ( 1977a) r epor t ed

t h e i r data i n d i c a t e d t h a t 0.001 of the LC50 a f f e c t e d

sheepshead minnows d e t r i m e n t a l l y . A 96 hr LC50 f o r a d u l t

sheepshead minnows was shown to be 70 ug/1 of Kepone. In 28

day chron ic exposure t e s t s , the a d u l t s died [ 22. 2%) from

c o n c e n t r a t i o n s as low as 0 . 0 8 ug / 1 .

The d i f f e r e n t l e v e l s of response could be due to the

f a c t t h a t tonsa responds to i t s sur roundings r a p i d l y and

i s more e n v i r o n m e n t a l l y a d a p t a b l e . Fac tors not ye t

i n t r oduced t h a t might modi fy r e s u l t s ob t a i ned here are the

i n c r e a s i n g of the l engt h of chron ic exposure , the use of


20

females and males s e p a r a t e l y , i n s t e ad o f randomly mixed

a d u l t s , and the use of a wide range of t empera tures as

opposed t o constant t e m p e r a t u r e . Perhaps in f o l l o w up

s t u d i e s i t can be determined i f chron ic exposure t ime should

be l e ng t h e n e d . I t i s probable t h a t the males and c e r t a i n l y

developmental s t a t e s are a f f e c t e d by changes in the

envi ronment be f or e the females (Conover , 1 9 5 6 ) . The re fo r e

under t h i s assumption chron ic s t r e ss could show up in males

bef or e f e ma l e s . Because A tonsa i s e u r y h a l i n e , s a l i n i t y

changes should have no n o t i c a b l e a f f e c t , however tempera ture

has a profound a f f e c t on metabol ism and a c t i v i t y and thereby

could a l t e r the response to s u b le t h a l l e v e l s .

P r e l i m i n a r y examinat ion of the e f f e c t s of l i g h t showed

no s i g n i f i c a n t d i f f e r e n c e s in g r az ing exper iments between

the l i g h t and dark c o n t r o l s . F i l t e r i n g r a t e has the

dimensions of volume x t i m e - * and is a measure of

the volume of ambient medium conta i ng the number of c e l l s

eaten by one animal in a given t i m e . Use of t h i s term does

not imply t h a t a l l the p a r t i c l e s of any given type are

removed from t h i s w a t e r , or t h a t the volume of water passed

over the f i l t e r i n g appendages i s known (Edmondson and

Winberg, 1 9 7 1 ) . At a c o n c e n t r a t i o n of 10 ug/1 of

c h l o r o p h y l l a, mean f i l t e r i n g r a t e s ( s i m i l a r to the

c o n t r o l s ) ranged from 5 . 15 t o 5 . 4 4 m l ‘ copepod- * * day- .

These r e s u l t s are in keeping w i th the r e s u l t s obta ined by

Conover (1956) and Reeve and W a l te r ( 1 9 7 7 ) .


I

21

An approx imat i on of f eed i ng r a t e was der i ved from the

f i l t e r i n g r a t e by m u l t i p l y i n g F by the a r i t h m e t i c mean of Cg

and Ct r a t h e r than by CQ alone (Edmondson and Winberg,

1 9 7 1 ) . This f eed i ng r a t e has the dimensions of

cel 1s ‘ copepod- * ( Tab l e 1 1 ) , which is in agreement wi th

Goldman and Peavey ( 1 9 7 9 ) , these f eed i ng r at es could then be

conver ted to ugC*copepod"* ( Tab l e 1 2 ) . Using the average

dry weight of A. tonsa of 7 ug (Conover , 1956 and Reeve and

W a l t e r , 1977) the f ee d i n g r a t e was then put i n t o terms of

ugC’ mg dry wt _ 1 * day_ 1 .

According to Conover (1956) the a du l t carbon

requ i rement f o r _A. tonsa a t 20°C is approx i mat e l y 130 ug

C'mg dry w t " ^ *d ay " ^ . Der ived f eed i ng ra t es of 158-164

ugC*mg dry w t “ * * d ay ~ * i n d i c a t e A . tonsa was meet ing i t s

carbon r e qu i re me nt . However a t 0 . 046 ug/1 of Kepone f o r

t h r e e days of exposure carbon i n g e s t i o n was s i g n i f i c a n t l y

l owered . This i n d i c a t e s the p o s s i b i l i t y t h a t su b le t ha l

l e v e l s of Kepone (were exposure long enough) in ambient

wate rs could reduce the l e v e l of carbon i n t a k e to

d e t r i m e n t a l l e v e l s f o r t o n s a . In 1976 Kepone l e v e l s in

wat er t aken from the James R i ve r ranged from <0.01 to lOppb,

no r ecent l e v e l s have been publ i shed ( Su t a , 1 97 8 ) .

Fo l low up s t ud i es t h a t would be v a l u a b l e to the o v e r a l l

unders tand i ng of the e f f e c t s o f Kepone in the e s t u r a t i n e

envi ronment i nc l ude work on a s s i m i l a t i o n e f f i c i e n c y ,

r e s p i r a t i o n r a t e s , r e p r o d u c t i o n , and b ioaccumula t ion

f a c t o r s .

■Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.

22

Ameri can

Bahner ,

Bougi s ,

Conover ,

Dawsey,

Deevey,

Desaiah,

Dubin, E

BIBLIOGRAPHY

P u b l i c He a l th S e r v i c e . 1975. Methods f o r the C u l t u r e and Short Terms Bioassay of the Ca lanoid Copepod A c a r t i a t o n s a . I n : Standard Methods f o rthe Examinat ion of Water and Wastewater. 14th Ed. APHA-AWWA-WPCF. New York.

. H . , A . J . Wi lson, J . M. Sheppard, J . M. P a t r i c k , and L.R. Goodman, 1977. Kepone B i o c o n c e n t r a t i o n , Accumul at i on , Loss and T r a n s f e r Through E s tua r i ne Food Chains, Chesapeake Sc ience . 1 8 ( 3 ) : 2 9 9 - 3 0 8 .

. 1976. Mar ine Plankton Ec ol ogy . Nor th-Hoi 1 and Pub!si hing Co . , Amsterdam-Oxford. 3 5 5 p .

R . J . 1956. Oceanography of Long I s l a n d Sound.1 952-54 VI Biology of A. c l a u s i and _A. t o n s a . Bul l Bingham Oceanogr. ColTec" 1 5:1 5 6 - 2 3 5 .

•H. 1964. P e s t i c i d e Reference Standards of the Entomological Soc ie ty of Amer ica. B u l l e t i n of Entomological So. Amer ica. 1 0 ( 2 ) : 9 5 - 1 03.

.B. 1956. Oceanography of Long I s l a n d Sound. 1952- 54. V Zooplankton . B u l l . Bingham Oceanogr.Col 1e c t . 1 5 : 1 1 3 - 1 5 6 .

D . , R.B. Koch. 1975. I n h i b i t i o n of ATPases A c t i v i t y in Channel C a t f i s h Brain by Kepone (Trade Name) and i t s Reduct ion Product . B u l l e t i n of Envi ronmental Contaminat ion and T o x i c o l o g y .1 3 ( 2 ) : 1 5 3 - 1 5 8 .

G. , A.G. Durb in . 978 Length and Weight R e l a t i o n s h i p of A c a r t i a c l a u s i from N a r r a ga ns e t t Bay R . I . Limnology and Oceanogr. 2 3 : 9 5 8 - 9 6 9 .


23

Edmondson, W.T. and G. G. Winberg. ed. 1971. A Manual on Methods f o r the Assessment of Secondary P r o d u c t i v i t y in Fresh Wat er s . B i a ck w e l 1 S c i e n t i f c Publ . Oxford .

Goldman, J . and D.C. Peavey. 1979. Steady S t a t e Growth and Chemical Compostion of the Mar ine Chlorophyte D u n a l i e l l a t e r t i c l e c t a in N i t rogen L i mi ted C u l t u r e s . Appl i ed and Envi ronmental Microb . 894- 901.

Gonzales, J . G . 1974. C r i t i c a l Thermal Maxima and Upper Letha l Temperatures f o r the Ca lano id Copepods A c a r t i a tonsa and A. c l a u s i . Mar ine B i o l ogy .2 ? : 2 1 9 - 2 2 3 . “

G u i l l a r d , R.L 1975 . C u l t u r e of Phytoplankton f o r Feeding Mar ine I n v e r t e b r a t e Animals . R e pr in t e d From: Cu l t u r e of Mar ine I n v e r t e b r a t e A n im a l s . Ed W.L. Smith and M.H. Chanley. Plenum P u b l i s h i n g Co. Mew York 2 9 - 6 0 .

Hansen. D . J . , A . J . Wi lson, W.R. Nimmo, S.C. Schimmel , L.H. Bahner . 1976. Kepone: Hazard t o Aquat icOrganisms. Sc ience . 1 9 3 ( 4 2 5 3 ) : 5 2 8 .

Hansen. D . J . , L . R . Goddman, A . J . Wi l son . 1977a. Kepone: Chronic E f f e c t s on Embryo. F r y , J u v e n i l e , and Adul t Sheepshead Minnows (Cypr inodon v a r i e g a t u s ) . Chesapeake Sc ience . 1 8 ( 2 ) : 2 2 7 - 2 3 2 .

Hansen, D . J . , D.R. Nimmo, S.C. Schimmel , G.G. Walsh, A . J .Wi l son . 1977b. E f f e c t s of Kepone on E s t ua r i ne Organisms. I n : Recent Advances in Fish Toxi cology . Envi ronmental P r o t e c t i o n Agency, E c o lo g ic a l Research S e r i e s . EPA.6 0 0 / 3 - 7 7 - 0 8 5 . : 20 - 30.

J e f f r i e s , H.D. 1962 . Succession of Two A c a r t i a Species in E s t u a r i e s . Limn, and Oceanogr. 7 : 3 5 4 - 3 8 4 .

K h a t t a t , F .H. and F a r l e y . 1976. Mar ine B i o t o x i c i t y ofC e r t a i n P e s t i c i d e s to A c a r t i a t o n s a ( D a n a ) . E.P. 1 . 2 3 : 6 0 0 / 3 - 7 6 - 0 3 3 .

Lance, J . 1963. The S a l i n i t y To l erance of Some E s t ua r i ne P l a n k t o n i c Copepods. Limn, and Oceanogr. 8 : 4 4 0 - 449.

Lance, J . 1965. R e s p i r a t i o n and Osmotic Behavior of theCopepod A c a r t i a tonsa in D i l u t e d Sea Water . Comp. Biochem. P h y s i o l . 1 4 : 1 5 5 - 1 6 5 .


24

Landry, H.R. 1976. The R e l a t i o n s h i p Between Temperature and the Development of L i f e Stages of the Marine Copepod A c a r t i a c l a u s i ( G e i s b r . ) . Limn, and Oceanogr . 2 0 : 8 5 4 - 8 5 7 .

L i t c h f i e l d , J . T . , F. Wi lcoxon. 1949. A S i m p l i f i e d Method of E v a l u a t i n g Dose E f f e c t Exper iments . J . Pharm. Exper . T h e r . 9 6 ( 2 ) : 9 9 - 1 1 5.

Mackenthun, K . M . , M.W. Broosman, J . A . K o h l e r , C.R.T e r r e l l . 1977 . M i t i g a t i o n F e a s i b i l i t y f o r the Kepone Contaminated James R i v e r , V i r g i n i a . In: Management of Bottom Sediments Cont a i n i ng Toxic Subst ances ; Proceedings of the T h i r d U . S. -Japan E x p e r t ' s Mee t ing . 1978. E P A - 6 0 0 / 3 - 7 8 - 0 8 4 . 153-181 .

M i l l e r , C . B . , J . K . Johnson, D.R. H e i n l e , 1977. Growth Rules in the Mar ine Copepod. Genus A c a r t i a . Limn, and Oceanogr . 2 0 : 3 2 6 - 3 3 5 .

P a r r i s h , K . K . , D .F . Wi lson . 1978. Fecundi ty Studies onA c a r t i a tonsa (Copepoda: C a l a n o i d a ) in Standard ized C u l t u r e . Mar ine B i o l o g y . 4 6 : 6 5 - 8 1 .

Reeve, M . R . , M.A. W a l t e r . 1977. Food Thresholds in A c a r t i a . J . Exp. B i o l . E c o l . 2 9 : 2 1 1 - 2 2 1 .

S a rg e n t , J . R . 1976. The S t r u c t u r e , Metabol ism and Funct ion of L i p i d s in Mar ine Organ isms. I n : Bi ochemi caland B i oph ys i ca l P e r s p e c t i v e in Mar ine B i o l o g y . Ed. D.C. Mal ins and J . R . S a r g e n t . Academic Press . New York . 447 p.

Schimmel , S . C . , A . J . Wi lson. 1977. Acute T o x i c i t y of Kepone to Four Es t u a r i n e Animals . Chesapeake S c i en c e . 1 8 ( 2 ) : 2 2 4 - 2 2 7 .

Schimmel , S . C . , M. P a t r i c k , L . F . Faas, J . L . Oglesby,A . J . W i l s o n . 1 979 Kepone: T o x i c i t y and Bi oaccumul at i on in Blue Crabs. E s t u a r i e s . 2(1 ) : 9 - 15.

S e k i g u c h i , H . , I . A . McLaren, C . J . C o r k e t t . 1 980. TheR e t a t i o n s h i p Between Growth Rate and Egg Product ion in the Copepod A c a r t i a c l a u s i hudsonica. Mar ine B i o l o g y . 5 8 : 1 3 3 - 1 3 8 .

S o k a l , R.R. and F . J . R o h l f . 1 969 . Bi o m et r y . W.H. Freeman and Company, San F r a n c i s c o . 766p.

S t e r r e t t , F . S . , C.A. Boss. 1977. Care less Kepone. Envi ronment . 1 9 ( 2 ) : 3 0 - 3 7 .


25

Stua, B.

W at ts , R

W i l l i ams

Wyman, K

. 1 978. Human Popu la t i on Exposures to Mi rex and Kepone. EPA 6 0 0 / 1 - 7 8 - 0 4 5 . 139pp.

R. et a l . 1980. De t er mi na t ion of Kepone in Human Blood and Envi ronmental Samples. In: Manual ofA n a l y t i c a l Methods f o r the Ana lys is fo P e s t i c i d e s in Human and Envi ronmental Samples. ETD, HERL, EPA.6 0 0 / 5 - 8 0 - 0 3 8 . Research T r i a n g l e p a r k , N.C.

D.A 1971 . A Test f o r D i f f e r e n c e s Between Treatment Means When Severa l Dose Levels Are Compared With A Zero Dose C o n t r o l . B i omet r i cs 27:1 03-1 1 7.

D . , H.B. O'Connors. 1980. I m p l i c a t i o n s of Short Term PCB-Contaminated Water , I n o rg an i c Sediments and P h y t o p l a k t o n . E s tu a r i n e and Coastal Mar ine Sc ience . 1 1 ( 2 ) : 1 2 1 - 1 3 1 .


26

Table 1. LIST OF REGISTERED PRODUCERS OF KEPONE PRODUCTS (1976) (SUTA, 1978 ) .

Company Address*

Act ion Product Corp. North Miami, FLA l l i e d Chemical Co. Morr i stown, NJAthena Corp. D a l l a s , TXBlack Leaf Products Co. E l g i n , ILBlack Magic Co. J a c k s o n v i l l e , flBoyle-Midway, I n c . C r a n fo r d , NJBrown Chemical S p e c i a l t y San Anton io , TXClarence Boord and Sons, I nc . Leon, IACommon Sense Manufactur ing Co. , I nc . B u f f a l o , NYF. C. S t u r t e v a n t Co. Cromwel1, CTGaston Johnson Corp. Long I s l a n d , NYGeneral Pest Se rv ice Co. Los Angeles, CAGrant L a b o r a t o r i e s D i v . , Le isure

E n t e r p r i s e s , I n c . Oakland, CAJ . & F. Manuf act ur ing Co. Houston, TXJohn O p i t z , I n c . Long I s a l n d , NYJudd Ringer Corp. Eden P r a i r i e , MNL e t h e l i n Products Co . , I n c . Mount Vernon , NYMcCal l Manuf ac t ur ing Co. Jas per , FLNott Manuf ac t ur ing Co. , I n c . P l easant V a l l e y , NY0. E. Linck D i v i s i o n , Walco Link Corp. C l i f t o n , NJOld 97 Company Tampa, FLPearson and Company M o b i l e , ALQuinn, Drug and Chemical Co. Greenwood, MSTrager Manuf ac t ur ing Co . , I nc . Scranton , PAVinson Chemical Products Co. Opalocks, FLWinn Chemical Co . , I nc . B I o u n t s v i 11e , al

Asgrow F l o r d i a Co. + P l an t C i t y , flChem Pack CO. + South Miami , FLLandia Chemical Co. + Lakeland, FLSouthern M i l l Creek Products Co. + Tampa, FL

♦Corporate address and not n ec e s s a r i l y manufactur ing add re ss .+These f our companies are s t a t e a p p l i c a n t s f o r federal regi s t r a t i on.Source: EPA, Federal R e g is t e r 4 1 ( 5 9 ) . (March 25 , 19 7 6 ) .


27

Table 2. Average Envi ronmental Kepone Co ncent ra t ions ( S u t a , 1 97 8 ) .

Source Average C onc en t r a t ion

James R i v e r

f i n f i s h 0 . 4 - 2 . O p pm

crabs 0 . 3 - 3 . 0 ppm

o ys t e r s 0.1 - 0 . 2 ppm

Chesapeake Bay

f i n f i s h 0 . 0 - 0 . 08 ppm

crabs 0 .10 - 0 . 26 ppm

o ys t e r s 0 . 008 - 0 .05 ppm


28

Tab le 3.

Specie

B l u e f i s h

Croaker

Grey t r o u t

Flounder

Spot

Tota l

Kepone in Chesapeake Bay Fish ( S u t a , 1 9 7 8 ) .

Sample AverageSi ze Kepone (ppm)

412 0 .03

249 0 . 06

106 0 . 05

45 0 . 08

86 0 .03

898 0 . 04


29

Table 4 . Kepone in Janies R i ver Fish ( S u t a , 1 9 7 8 ) .

Speci eSampli ng Locat i on

Sample Si ze

Average Kepone (ppm)

Channel c a t f i s h Unspeci f i ed 205 0 . 04

White C a t f i s h Unspeci f i ed 32 0 . 16

Largemouth bass Unspeci f i ed 36 1.12

Black c r ap p ie Unspeci f i ed 23 1 .03

Shad Zone 0 26 0 . 02

Zone A 34 0 . 0 2

Zone B 35 0 . 02

Zone C 23 0 . 07

Weyonoke 10 0 . 12

Al l Zones 1 28 0 . 0 4

B I u e f i sh Unspeci f i ed 23 0 . 65

Spot Unspeci f i ed 16 0 . 75

Croaker Unspeci f i ed 33 0 . 62

S t r i p ed m u l l e t Unspec i f i ed 4 0 . 37

S t r i p e d bass Unspeci f i ed 8 0 . 04

Grey t r o u t Unspeci f i e d 4 1 .21

Speckled t r o u t Unspeci f i ed 8 0 .07

American eel Unspeci f i ed 16 2 . 37

Oyster toad Unspeci f i ed 3 2 .53

Hogchoker Unspeci f i ed 18 0 . 93

Whi te perch Unspeci f i ed 31 2.06


30

Table 5. C a l c u l a t i o n o f Kepone Concent ra t ions Recovered by EC - GC.

1) Kepone c o n c e nt r a t i on from G.C. read out in pg = x pg

2) x pg -f- i n j e c t i on vol ume = xpg/ul in in j e c t i on vol ume

3) xpg /u l x ul in t o t a l e x t r a c t = xpg in e x t r a c t

4) xpg in e x t r a c t -f- l i t e r s of water sample e x t r a c t e d =xpg/1 in water


31

Table 6. C a l c u l a t i o n s f o r F i l t e r i n g and Feeding Rates .

G = graz i ng c o e f f i c i e n t a

K = growth c o e f f i c i e n t b

CQ = i n i t i a l c e l l number

Ct = c e l l number at t ime t

t = exper iment a l t ime

V = volume ambient medium

N = number of copepods

F = f i l t e r i n g r a t e c

I = f eed i ng r a t e d

aG = InCo - InCt + Kt t

bK = InCt - InCo t

CF = V x G N

rt Co + CtI = F x ---------------

2


32

Tab le 7. Resu l ts of EC-GC Kepone An a lys is from C o l l e c t i o n S i t e .

C onc en t r a t ion C onc en t r a t ion %Added Recoveredug/1 ug/1

Mi rex Spike 0 . 8 0 . 83

Kepone Standci rd 1 .6 1 .46

Water Sample 1 Unknown BDL*



*Below d e t e c t i o n l i m i t s o f 0 . 0 2 ug/1 of Kepone.

Recovery

103

91 .25


Table 8 . A c a r t i a tonsa Acute T o x i c i t y Bioassay M u l t i p l e Range Test -Duncan Procedure f o r the 0 . 05 l e v e l

Homogeneous Subsets Nominal Kepone Mean M o r t a l i t y( u g / 1 ) ( Arcs i ne-fP* t ra n s .

1 500 .0 90 . 00

2 50.0 75 .00

3 5 . 0 54 . 78

4 0.5 33 .00

5 0 .05 0 . 00


34

Table 9.

Homogeneou s

1

D u n a l i e l l a t e r t i o l e c t a Chronic T o x i c i t y T e s t . M u l t i p l e Range Test-Duncan Procedure f o r the 0 . 0 5 l ev e l

Subsets Nominal Kepone ( u g / 1 )

Mean Cel 1 Number

0.046 7810 .000

0.0046 7883.333

0 . 00 7926.667


35

Table 10.

Homogeneous

1

2

A c a r t i a tonsa Grazing Exper iment under Chronic Exposure. M u l t i p l e Range Test -Duncan Procedure f o r t he 0 . 0 5 l e ve l

Subsets Nominal Kepone Mean F i l t e r i n g( u g / 1 ) Rate

m l ‘ copepod- 1 ‘ day-1

0 .046 4 . 63

0 .025 5 .15

0 . 000 5 .35

0 . 0046 5 .44


36

Table 11. Data from carbon a n a l y s i s of D u n a l i e l l a t e r t i o l e c t a c u l t u r e averaged from t h r e e r e p l i c a t e s .

i * i bu l C02 * 5ml sample-1 a pgC * c e l l

1 7 . 2 30( 3 . 4 u l C02 *ml - i )

a Each 5 ml sample conta ined 73000 c e l l s * ml

b There a r e 6 . 357 x 10- gC * u l C0 2- ^


37

Table 12. Derived Fi l ter ing and Feeding Rates of Acartia tonsa on Puna!iel la t e r t i o l ec ta .

Nominal Mean F ilte r in g Rate Mean Feeding RageKepone (ug/1) ml•copepod_1*day" ugC'copepod day ugc'mg dry wt-1

0.046 4.63 1.641 150

0.025 5.15 1.805 158

0.0046 5.44 1.906 165

0.0000 5.35 1.896 164


38

Table 13. Computer LCjq and LC^q Values with 95% Confidence Limites fo r 96-Hour Exposure Acartia Tonsa

Pesticide

methyl parathion* (80% Technical)

Azodrin*(80% Technical)

Kepone(94.4% Technical)

diazinon*(97.6% Technical

toxaphene*(100% Technical)

* (Khattat and Farley

0.07 ppm(0.0418 - - / ; ;45 ppm)

0.05 ppb(0.0098 - 0.269 pb)

0.115 ppb (0.0235 - 0.465)

0.04 ppb(0.0164 - 0.0777 ppb)

0.0035 ppt(0.0006 - 0.0187 ppt)

1976)

0.089 ppm(0.685 - 1.163 ppm)

0.24 ppm(0.08536 - 0.66170 ppm)

4.96 ppb(2.610 - 9.433 ppb)

2.57 ppb(1.7259 - 3.8247 ppb)

7.2 ppt(3.540 - 14.636 ppt)


Kepone

CL

CL

Cl''C '

C L

*— - (

*V

’'C l

% CS/ \' 'C l

M ire x

/'Cl

'#0

Cl

-Cl

C L

\'C l

C LO “

t \^ ~ ' C I

Cl

Cl

Figure 1. Similar Molecular Strucutres of Kepone and Mirex.


Reproduced

with perm

ission of the

copyright ow

ner. Further

reproduction prohibited

without

permission.

JORDAN POINT COUNTRY CLUB/

ALLIEDCHEM. BAILEY

BAYHOPEWELL Industrial 0 Wasto

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SCALE IN MILES

Figure 2. Hopwell, Virginia and Bailey Bay James River (From U.S. Geological Survey, Washington, D.C.)

o

Reproduced

with perm

ission of the

copyright ow

ner. Further

reproduction prohibited

without

permission.

QUEEN CR.

WILLIAMSBURGHOPEWELL

’CLAREMOI

SEDIMENT CONCENTRATIONS ppm Kepone

New po rtF: NEWSi;

SMITH FI ELD

Figure 3. James River Sediment Concentrations from Hopewell, Virginia to Newport News, Virginia, August 1976 (Mackenthun et al., 1977)

-I*

42

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LOOK TR

- I 20»Atf-vls - ‘ TV CAP q Mo (U) 20se 164ft ISM

HORN R Bn 298-------H E N R Y

MCR CL. 84 FTVERT CL. 35 FT

OVHO PWR. CAB AUTH.CL.88 FT

I

\ £ ><4l23««c IS 't ~ ' 3

Alv-\1 .

6 ± J f lW « 5 £Z

Figure 4. Collection Site at Long Creek Bridge.Source. Williams and Neintz Map Corporation, 1977.


the effects of kepone on the estuarine copepod acartia tonsa

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