United States                        SR-111
           Environmental Protection
           Agency
EPA       Research  and
           Development
           ERLGB PUBLICATIONS AND ABSTRACTS
           RELATED TO BIOTECHNOLOGY!

             E. BIORATIONAL, MICROBIAL AND
                BIOCHEMICAL CONTROL AGENTS
           Prepared by

           Environmental Research
           Laboratory
           Gulf Breeze FL 32561
                                 February, 1984

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COUCH,  J.A.,  S.H.  MARTIN, G, TOMPKINS, AND J.  KINNEY.  IN  PRESS.  SIMPLE
SYSTEM  FOR  THE PRELIMINARY EVALUATION OF INACTIVITY  AND  PATHQGENESIS  OF
INSECT VIRUS IN A NONTARGET ESTUARINE SHRIMP.  J. INVSRTEBR. PATHOL.  <£RL,GB
460).

   BIOLOGICAL CONTROL AGENTS (BIORATIONALS) ARE INCREASINGLY IMPORTANT IN
   PEST  CONTROL  CONCEPTS.  CERTAIN  INSECT  VIRUSES,  PARTICULARLY  THE
   BACULOVIRUSES  {NUCLEAR PCLYHEDROSIS VIRUSES), ARE CONSIDERED TO  HAVE
   POTENTIAL  AS BIOLOGICAL PESTICIDES, AND COULD BE USED WIDELY  IN  THE
   ENVIRONMENT-  THEREFORE,  TEST ANIMALS MUST BE  SELECTED  AND  METHODS
   DEVELOPED  TO  EVALUATE  THE  SAFETY  OF  BIORATIONALS  TO  NON-TARGET
   SPECIES.  A SIMPLE LABORATORY SYSTEM HAS BEEN DESIGNED AND  TESTED  TO
   DETERMINE RISKS OF INFECTIVITY AND PATHOGENICITY OF AN INSECT
   BACULOVIRUS,  ORIGINALLY ISOLATED FROM THE ALFALFA  LODPE8  AUTOGRAPHA
   CAILFORNICA, TO A NON-TARGET ARTHROPOD, THE GRASS SHRIMP, PALAEMONETES
   VULGARIS,  BY  DIETARY EXPOSURE. THIS LABORATORY METHOD  ALSO  PERMITS
   TESTING  OF  OTHER MICROBIAL BIORATIONALS AGAINST  NON-TARGET  AQUATIC
   SPECIES,  AND PROVIDES AND INEXPENSIVE, PROCEDURE OF  SAFETY  TESTING.
   RESULTS PROM THIS STUDY INDICATED THAT HISTOPATHOLOGICAL,
   ULTRASTRUCTURAL AMD SEROLOGICAL METHODS US50 PROVIDED NO EVIDENCE THAT
   EXPERIMENTAL  EXPOSURE TO THE VIRUS CAUSED VIRAL INFECTION OR  RELATED
   PATHOGENICITY IN THE GRASS SHRIMP.

COUCH,  JOHN.  1975.  DISCUSSION FROM SELECTED PAPERS PRESENTED  AT  EPA-USOA
WORKING SYMPOSIUM.  IN:  BACULOVIRUSES FOR INSECT PEST CONTROL:  SAFETY
CONSIDERATIONS.  MAX  D. SUMMERS, RILO ENGLER, LOUIS A. FALCON, AND P.  VAIL,
EDITORS, AMERICAN SOCIETY FOR MICROBIOLOGY, WASHINGTON, DC.  PP.
58-62,111-114.  
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COUCH,  JOHN A.  1974.  ENZOOTIC NUCLEAR PQLYHEDROSIS  VIRUS  OF   PINK   SHRIMP:
INFRASTRUCTURE, PREVALENCE, AND ENHANCEMENT.  J,  INVERTEBR.  PftTHOL,
24C3>:311-331.  CERL,GB 215),

   A NUCLEAR POLYHEDRCSIS VIRUS EXISTS  IN PINK  SHRIMP,  PBH&EUS   QUGRARUM,
   FROM WATERS OF THE NORTHERN GULF OF  MEXICO.  THIS VIRUS  IS ROD-SHAPED,
   269 NM LONG, AMD POSSESSES AN OUTER  ENVELOPE SURROUNDING  ITS
   NUCLEOCAPSIO. THE NUCLEOCAPSID IS 50 NM IN DIAMETER. THE  VIRUS  OCCURS
   IN NUCLEI OF HOST HEPATOPANCREATIC AND MIOGUT  CELLS, AND  IS 8QTH   FREE
   IN THE NUCLEUS AND OCCLUDED WITHIN PYRAMIDAL-SHAPED  POLYHEDRAL
   INCLUSION  BODIES (PIB'S). HISTOCHEMICALLY AND ULTSASTRUCTRUALLY,   THE
   SHRIMP PIB'S APPEAR TO BE RIBONUCLEOPROT2IN  AND IN  FINE STRUCTURE  BEAR
   CLOSg  RESEMBLANCE  TO  POLYHEDRAL   INCLUSION  BODIES   OF BACULOVIRUS
   SPECIES  FROM  INSECTS. HOWEVER, THE LATTICE LINE-TO-LINE SPftCINS  IS
   GREATER  THAN  THAT USUALLY REPORTED FOR INSECT  PIE'S.   CR08DIN6   AMD
   CHEMICAL  STRESS  OF SHRIMP IN AQUARIA MAY ENHANCE   AND   INCREASE   THE
   VIRUS  INFECTION  AND PREVALENCE. IN LIMITED EXPERIMENTS,  SHRIMP   FED
   HEAVILY  INFECTED HEPATOPANCREATIC TISSUES HAD MUCH  HIGHER   MORTALITY
   THAN CONTROLS FED ONLY FISH. THE VIRUS APPEARS TO BE ENZOOTIC IN   PINK
   SHRIMP IN NATURE. CVTOPATHOLOGICAL CHANGES IN  INFECTED CELLS  OF SHSIMP
   APPEAR  SIMILAR TO THOSE IN INSECTS  INFECTED WITH CERTAIN  SPECIES   OF
   6ACULOVIRUS.  TH£  NAME BACULOVIRUS  PENAEI N.SP. IS  PROPOSED  FOR   THE
   SHRIMP VIRUS.

COJJtH,  JOHN A.  1974,  FREE AMD OCCLUDED VIRUS,  SIMILAR TO  BACULOVIRUS,   IN
HEPATOPANCREAS  OF PINK  SHRIMP.  NATURE.  247C5438):229-231.  CERL,GB  213).

   A  ROD-SHAPED,  FREE  AND OCCLUDED VIRUS EXISTS IN   A   MARINE  SHRIMP,
   INDICATING  THAT  MARINE  CRUSTACEA  ARE POTENTIAL   HOSTS  FOR  VIRUSES
   SIMILAR  TO CERTAIN VIRUSES INFECTING INSECTS  AND MITES*  SO   FAR,   THE
   VIRUS  HAS  BEEN FOUND ONLY IN SHRIMP TAKEN FROM NEAR   CEDAR  KEY   AND
   EXPERIMENTALLY EXPOSED TO TH£ TOXIC CHEMICAL,  AROCLOR 1254 CPC8).   THE
   VIRUS PROBABLY IS A NATURAL PARASITE, HOWEVER, PREVIOUSLY UNDETECTED,
   OF  cSTUARINc AND MARINE SHRIMP. STUDIES OF  POSSIBLE  INTERACTIONS   OF
   THE  PCS  AND VIRUS IN PINK SHRIMP MAY  PROVIDE  VALUABLE INFORMATION
   NEEDED TO CLARIFY THE RELATIONSHIP BETWEEN NATURAL  INFECTIOUS DISEASES
   AND POLLUTANT CHEMICALS IN THE AQUATIC ENVIRONMENT.
                                 PAGE

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COUCH,  JOHN A.  1974,  PATHOLOGICAL  6FFECTS OF  URCSPQ"RIOIUM     LOSS OF MOBILITY
   AND  RESISTANCE  TO MECHANICAL PRESSURES. THOUGH  SEVERE PATHOLQSICftL
   CHANGES OCCUR IN HEAVILY INFECTED  METACERCARIAE,  HOST  INFECTS®
   HETACERCARIAE REMAIN VIABLE WITHIN THE BLUE CRAB  AND  THERBY  SERVE  AS A
   VECTOR FOR URQSPQRIDIUM UNTIL THE  DEATH OF THE BLUS CRAB. AT  THE   TIME
   OF  TH£  CRAB'S  DEATH  AND  DISORGANIZATION,  INFECTED   METACERCftRIAE
   RUPTURE AND RELEASE SPORES OF THE  HYPERPARASITE.

COUCH,  JOHN A.  1975.  VIRUS FROM PINK  SHRIMP (ABSTRACT).   PRESENTED AT  THE
FIRST WORKSHOP ON THE PATHOLOGY AND TOXICOLOGY OF PENAEID SHRIMP, APRIL 8-10,
1975, GALVESTON, TX.  (ERL,GB X291).

   SINCE  THE REPORT OF A BACULQVIRUS FOUND IN PINK  SHRIMP  APPEARED   OVER
   OHc YEAR AGO, CONSIDERABLE INFORMATION HAS ACCUMULATED ON THE
   VIRUS-SHRIMP RELATIONSHIP. THE FOLLOWING TENETS APPEAR TO HOLDS <1)   A
   BACULOVIRUS (NUCLEAR POLYHSDROSIS  VIRUS) INFECTS  PINK  SHRIMP  NATURALLY
   YEAR  ROUMD  IN  CERTAIN AREAS OF  THE GULF OF HEXICOJ  (2)   THE  VIRUS
   CAUSES CONSIDERABLE CYTOPATHOLOGY  IN  HEAVY INFECTIONS  OF THE
   HEPATOPANCREAS;  (3)  THE VIRUS IS ENZOOTIC AND MAY  BECOME   EPIZOOTIC
   UNDER  CERTAIN STRESS CONDITION TO WHICH THE HOST IS  EXPOSED:  AND  (4)
   SHRIMP ARE SIMILAR TO OTHER ARTHROPODS IN BEING HOSTS FOR AT  LEAST ONE
   VIRUS.

COUCH, JOHN A.  1976.  ATTEMPTS TO INCREASE BACULOVIRUS  PREVALENCE IN   SHRIMP
BY CHEMICAL EXPOSURE.  IN:  TUMORS IN AQUATIC ANIMALS.   CLYDE J.  OAWE,   DANTE
G. SCARPELLI, AND SEFTON R. SELLINGS, EDITORS, S. KARGER, BASEL.  20S304-314.
<6RL,G8 240).

   LITTLE INFORMATION IS AVAILABLE CONCERNING INTERACTIONS  BETWEEN
   POLLUTANT  CHEMICALS AND VIRUSES IN AQUATIC ANIMALS.  S&MPLES   OF   PINK
   SHRIMP  (PENAEUS DUORARUM) WITH VARIOUS ENZOOTIC  LEVELS  OF  A  NATURAL
   BACULOVIRUS  INFECTION  WERE EXPERIMENTALLY EXPOSED TO LOW  LEVELS   OF
   AROCLOR  1254, A POLYCHLORINATED BIPHENYL CPCB) ,  MIR6X,   CADMIUM,  AND
   METHQXYCHLOR  IN THE LABORATORY. NO CONSISTENT PATTERN OF INCREASE   IN
   PREVALENCE  OF VIRUS WAS FOUND, AND NO INDICATION OF  TUMOR   INDUCTION
   WAS DETECTED.
                                 PAGE

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COUCH,  JOHN A,  1977.  INTERACTION OF THREE PATHOGENS IN MASS  MORTALITY   OF
PENAEIO SHRIKP 
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COUCH, JOHN A.  1978.  DISEASES,  PARASITES, AND TOXIC  RESPONSES OF COMMERCIAL
PENAEID  SHRIMPS  OF THE  GULF  OF  MEXICO AND SOUTH ATLANTIC   COASTS  OF  NORTH
AMERICA.  
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COUCH, JOHN A.  1979.  SHRIMPS (ARTHRQPQDA:  CRUSTACEA:  PENAEIQAE).  IN:
POLLUTION  ECOLOGY  0*=  ESTUARINE INVERTEBRATES.  C.W.  HiRT,  JR..  AND  L.H.
SAMUEL, EDITORS, ACADEMIC PRESS, NEW YORK, NY.  PP. 235-258.  
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COUCH, JOHN A., AND LEE COURTNEY.   1977.   INTERACTION  OF  CHSMICAL   POLLUTANTS
AND  VIRUS IN A CRUSTACEAN:  a NGVEL  3IOASSAY  SYSTEM.   ANN.  N.Y.   ACAD.   SCI.
298J497-5S4.     AT
   0.7  PP8  FOR  35  DAYS  IN FLOklMG  SEAtfATER.  THE  OTHER   GROUP   WAS
   MAINTAINED AS A CONTROL GROUP IN FLOWING SSAWATER,  VIRAL  PREVALENCE It*
   EXPOSED SHRIMP SAMPLES INCREASED WITH  TIME  AT A SIGNIFICANTLY   GREATER
   RATE  THAN DID VIRAL FREQUENCY IN  CONTROL SHRIHP. VIRAL PREV&LiMCE   IM
   AROCLOR-EXPOSEO  SHRIMP  SURVIVORS WAS  751  AFTER 35 DAYS,   WHEREAS   XN
   CONTROL  SHRIMP, ONLY *5.?S HAD  PATENT  VIRAL INFECTIONS.  THIS   FILING
   SUGGESTS AN INTERACTION AMONG CHEMICAL  STRfiSSOR CAROCLQR  1254),  HfiST,
   AND  VIRUS. THE NATURE OR MECHANISM OF  THIS INTERACTION HAS NOT  6EEH
   DEFINED, BUT THE SHRIMP-VIRUS SYSTEM SHOWS  PROMISE  FOR FUTURE
   8IDASSAYS OF INFLUENCE OF LOW CONCENTRATIONS OF POLLUTANTS  ON   NATURAL
   PATHOGEN-HOST INTERACTIONS.

COUCH,  JOHN  A., AND SUSAN MARTIN.   1982.  PROTOZOAN  SYMBIONTS  AND   RELATED
DISEASES OF THE BLUE CRAB, CALLINECTES SAPIDUS RATHBUN, FROM THE ATLANTIC AND
GULF  COASTS  OF  THE  UNITED  STATES.  INS  PROCEEDINGS  OF   THE  BLUE   CRAB
COLLOQUIM, 3ILOXI, MISSISSIPPI, OCTOBER 16-19, 1979.   HARRIET  M. PERRY AND W.
A.  VAN  ENGEL,  EDITORS,  GULF STATES  MARINE  FISHERIES  COMMISSION,   OCEAN
SPRINGS* MS.  PP. 71-80.  CERL,GB 325).

   THE-  BLUE CRAB CCALLINECTES SAPIDUS RATHBUH, lB9fe)  SUPPORTS  VALUABLE
   FISHERIES ALONG THE MIO-ATLANTIC AND GULF COASTS OF THE UNITED  STATES.
   BECAUSE  THE CRAB IS AN ESTUARINE  SPECIES,  CAPABLE  OF  RANGING  WIDELY
   WITHIN  ITS  HABITAT, IT IS SUBJECT TO  THE  RIGORS   OF  THE   EURYHALINE
   ENVIRONMENT, AS WELL AS TO THE STRESSES CAUSED BY HUMAN ACTIVITY ALONG
   COASTLINES, IT HAS BEEN DEMONSTRATED THAT CAPTIVE-CRAB POPULATIONS  ARE
   PARTICULARLY  SUSCEPTIBLE  TO PARASITES AND COMMENSALS, AND TO  THEIR
   ASSOCIATED DISEASE AND DEBILITIES. WITHIN WILD POPULATIONS,  EXTENSIVE
   MORTALITIES  DUE  TO THESE FACTORS ARE  DIFFICULT TO  MONITOR,   BUT  DO
   CAUSE  FLUCTUATING  LOSSES  TO THE CRAB FISHERY.  THIS  PAPER   REVIEWS
   EXISTING KNOWLEDGE ON THE MORE COMMON  PROTOZOAN SYMBIONTS AND DISEASES
   OF  THE  BLUE  CRAB FOUND ON THE EASTERN AND  GULF  COASTS.  AVAILABLE
   INFORMATION ON RECOGNITION AND DIAGNOSIS OF DISEASE, SITE OF
   INFECTION,  AND  PATHOGENICITY   IS INCLUDED.  DATA  ARE   PRESENTED  ON
   TAXONOMY, MORPHOLOGY, AND LIFE CYCLES  OF ASSOCIATED PROTOZOAN
   PARASITES AND COMMENSALS OF CALLIN6CTES SAPIDUS.

COUCH,  JOHN  A., AND D.R. NIMMO,   1974,   DETECTION OF  INTERACTIONS   BETWEEN
NATURAL PATHOGENS AND POLLUTANTS IN AQUATIC ANIMALS.   IN:  PROCEEDINGS OF THE
REGIONAL  SYMPOSIUM ON DISEASES OF  AQUATIC ANIMALS.  LSU-SG-74-05,  LOUISIANA
STATE  UNIVERSITY,  CENTER  FOR  WETLAND   RESOURCES,  BATON  ROUGE,  LA.   PP.
261-268.  
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COUCH, JOHN A., AND DELWAYNE R. NIMMO.  1973.  CYTQPATHGLOGY, ULTfcASTRUCTURE,
AND  VIRUS  INFECTION  IN  PINK  SHRIMP EXPOSED  TO  THi  PC8,  AROCLQR   1254
CABSTRAQT).  IN:  PROCEEDINGS  OF  THE  JOINT  MEETING  OF   THE  SOCIETY   FOR
INVERTEBRATE  PATHOLOGY AND INTERNATIONAL COLLOQUIUM 0« INSHCT PATHOLOGY   ANO
MICR08IAL CONTROL.  PP. 105.  (5RL,GB 242).

   LITTLE  INFORHATION IS AVAILABLE CONCERNING THE EFFECTS   OF  POLLUTANT
   CHEMICALS  ON THE FINE STRUCTURE OF TISSUES IM AQUATIC  INVERTEBRATES.
   EVEN  LESS  IS  KNOWN CONCERNING POSSIBLE  INTERACTIONS   OF  POLLUTANT
   CHEMICALS  AND NATURAL PATHOGENS IN VALUABLE INVERTEBRATE SPECIES.  IN
   EXPERIMENTS AT THE GULF BREEZE, FLORIDA EPA LABORATORY WE HAVE 6XPOSHO
   PINK  SHRIMP  CPENAEUS  DUQRARUM) TO 3 PPB  ARQCLQR  1254  IM  FLQUING
   S6AWATER FROM 30 TO 52 DAYS. DURING THESE EXPOSURES OP TO 50f OR  MORE
   OF  THE  ANIMALS  DIED. SAMPLES OF BOTH LIVING AftD  DEAD  SHRIMP  MERE
   ANALYZED FOR AROCLOR RESIDUES ANO, AFTER 30 DAYS EXPOSURE, WERE  FOUSD
   TO ACCUMULATE FROM 33 PPM TO 40 PPM IN THEIR HEPATOPANCREATIC TISSUES.

COUCH, JOHN A., AND K. RANGA RAO, EDITORS,  1983,  BIORATION&L WORKSHOP, GULF
BREEZE, FLORIDA, SEPTEMBER 15-17, 1982.  EPA-600/X-83-054, U.S. ENVIRONMENTAL
PROTECTION AGENCY, ENVIRONMENTAL RESEARCH LABORATORY, GULF BREEZE,  FL.  64P.

   THi GENERAL PURPOSE OF THE WORKSHOP MAS TO EVALUATE THE
   STATE-OF-THE-ART OF TESTING, AMD THE SAFETY OF BIORATIONALS TO  BIROS,
   MAHMALS, AQUATIC ORGANISMS, PLANTS, AND INSECTS ANO TO REVIEW THE PART
   OF  SU8PART  14 OF THE GUIDELINES CGUIOLINES DOCUMENT  FOR  REGISTERING
   PESTICIDES  IN  THE  U.S.:  BIORATIONALS) DRAFTED  BY  THE  ECOLOSICAL
   EFFECTS BRANCH, OFFICE OF PSTICIOES PROGRAHS .
                                 PAGE

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COUCH,  JOHN  A.,  MAX D. SUMMERS,  AND  LEE  COURTNEY.  1975.  ENVIRONMENTAL
SIGNIFICANCE  OF  BACULOVIRUS  INFECTIONS IN  ESTUARINE  &NO  MARINE  SHRIMP.
ANN.N.Y. ACAD, SCI.  266:528-536.  
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COUCHt  JOHN,  GEORGE GARDNER, JOHN C, HARSHBARGER,  M.R,  TRIPP,  AND  PAUL   P,
YEVICH.  1974,  HISTOLQGICAL  AND  PHYSIOLOGICAL  EVALUATIONS  IN   SOME  MARINE
FAUNA.  IN:  MARINE  8IOASSAYS.  MARINE TECHNOLOGY  SOCIETY,   WASHINGTON,   OC.
PP. 156-173.  C£RL,GB 250).

   THE  DEVELOPMENT  OF  PATHOLOGY, AS  APPLIED   TO   AQUATIC   TOXICOLOGY,
   DEPENDS HEAVILY ON THE KNOWLEDGE OF NORMAL HISTOLOGY AND PHYSIOLOGY  IF
   ANOMALIES, DUE TO POLLUTANTS OR DISEASE, ARE TO  BE  ACCURATELY  DEFINED.
   HOWEVER,  AT  PRESENT,  KNOWLEDGE OF NORMAL  MORPHOLOGY  OR   METABOLIC
   ACTIVITIES IS EITHER INCOMPLETE OR LACKING FOR MOST MARINE  OR   COASTAL
   ORGANISMS.  OBVIOUSLY,  WELL-COORDINATED EFFORTS  WILL  BE   REQUIRED  TO
   CHARACTERIZE NORMAL RANGES AND INTERPRET THE MORPHOLOGICAL  OR
   PHYSIOLOGICAL  RESPONSES  OF  AQUATIC  ORGANISMS   TO   VARIOUS   FACTORS
   INCLUDING  POLLUTANTS. MOST LABORATORIES CANNOT   JUSTIFY   WELL-DEFINED
   PATHOLOGICAL UNITS, ALTHOUGH THE NEED OFTEN ARISES  THROUGH
   GOVERNMENTAL  ENFORCEMENT  ORGANIZATIONS  AND  OTHER   ACTIVITIES.  THE
   TECHNIQUES  AND  EXAMPLES THUS PRESENTED ARE INTENDED  TO   OFFER   SOME
   MEANS  OF  OBTAINING EVALUATIONS OF AQUATIC ORGANISMS"  WELL-BEING  OR
   DISORDERS.

SPRAGUE, VICTOR, AND JOHN COUCH.  1971.  ANNOTATED LIST OF PROTOZOAN
PARASITES* HYPERPARASITES, AND COMMENSALS OF DECAPOD  CRUSTACEA.   J.
PROTOZOOL.  !8(3):526-537.  
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