Product Citations and Abstracts

In situ measurement of scleractinian coral size and structural complexity in has been elusive. Three-dimensional (3D) colony surface area has been measured in the laboratory but the techniques are inappropriate for underwater field surveys. Recent field studies have demonstrated the potential of 3D colony surface area measurements for evaluating coral condition. This potential would be enhanced by an ability to accurately estimate 3D coral surface area from rapid underwater field measurements. Here we apply non-destructive photographic methods to document size and structure of corals in the field. Multiple images of a single colony were used to generate a scaled 3D computer reconstruction of colony morphology that can be accurately measured for colony dimensions and 3D surface area. Methods were applied to 23 colonies of two stony coral species, Diploria clivosa and D. strigosa, photographed in situ at the Florida Keys and Dry Tortugas. Accuracy of the technique was examined by comparison with a laser-scanned laboratory colony, and precision was documented through repetitive model construction. Surface areas and volumes calculated from digitally reconstructed corals were compared to colony dimensions and to size classes assigned in the field. These comparisons provided a means to estimate coral surface area and volume from simple field measurements and demonstrated that surface area of Diploria colonies closely approximated hemispheric surface area. Further application of this technique to additional coral taxa will advance methodology for assessment of coral and coral reef condition as well as for understanding surface-related physiological processes of coral.

Courtney, Lee A. and John A. Couch. 1981. Aspects of the Host-Commensal Relationship Between a Palaemonid Shrimp (Pontonia domestica) and the Pen Shell (Atrina rigida). EPA-600/J-81-041. Northeast Gulf Sci. 5(1):49-54. (ERL,GB 249). (Avail. from NTIS, Springfield, VA: PB82-158171)

Sixty pen shells (Atrina rigida), were collected and examined to determine the prevalence and morphometrics of the commensal shrimp (Pontonia domestica). The spatial and sexual distribution within the shell of the host, and the reproductive states of the shrimp were also investigated.

Courtney, Lee A. and John A. Couch. 1984. Usefulness of Cyprinodon variegatus and Fundulus grandis in Carcinogenicity Testing: Advantages and Special Problems. EPA-600/J-84-063. Natl. Cancer Inst. Monogr. 65:83-96. (ERL,GB 442). (Avail. from NTIS, Springfield, VA: PB84-226513)

Cyprinodon variegatus and Fundulus grandis, two species of cyprinodontid fishes extensively studied and used in toxicological and biological investigations, are compared as laboratory test animals. Their ecology, general biology, and suitability for various types of experimentation are examined. A laboratory system for exposure of these species at critical life stages (embryos, fry, juveniles) to suspect carcinogens is described. A discussion of the use, findings, and potential of these species in oncological studies and carcinogen assays is presented, particularly in regard to responses to 3 known or suspect carcinogens (trifluralin, benzidine, and aflatoxin). Finally, advantages and disadvantages or special problems in using the species as carcinogen test animals are reviewed.

Courtney, Lee A. Unpublished. Rodlet Cells in Fishes. In: Pathobiology of Spontaneous and Induced Neoplasms in Fishes: Comparative Characterization, Nomenclature, and Literature. Clyde J. Dawe, John C. Harshbarger, Sefton R. Wellings, and John D. Strandberg, Editors. Academic Press, Orlando, FL. 64 p. (ERL,GB 623).

The rodlet cell is an enigmatic cell type that has provoked much controversy among parasitologists and histologists since its first description (Thelohan, 1892a, b). It has frequently been associated with neoplasms and other pathological conditions (Dawe et al., 1964; Anderson et al., 1976; Hawkins, 1984; Smith, 1984; Mitchell, 1989) as well as in ostensibly normal tissues in many species of teleost fish. Furthermore, recent experimental studies have demonstrated extensive accumulations of rodlet cells associated with development of hepatic neoplasms in the sheepshead minnow, Cyprinodon variegatus, following exposure to N-nitrosodiethylamine (DENA) (Couch and Courtney, 1987). In view of these associations, the controversial nature of rodlet cells, and the likelihood that investigators of fish neoplasms will encounter this cell type, an overview, description and discussion of rodlet cells in teleosts is relevant to this volume.

Courtney, Lee A. and John W. Fournie. 1991. Ocular Chondrosarcomas in Rivulus marmoratus Poey. EPA/600/J-91/101. J. Fish Dis. 14(1):111-116. (ERL,GB 679). (Avail. from NTIS, Springfield, VA: PB91-206946)

Only one documented case of a malignant cartilaginous tumor has been previously reported from fish, a large cranial chondrosarcoma from the paddlefish, Polyodon spathula (Walbaum). Described here are two chondrosarcomas diagnosed from adult laboratory-reared Rivulus marmoratus Poey, the first reports of this tumor type from an atheriniform fish. Histologically, the tumors consisted of cartilage in various stages of differentiation. These ranged from poorly differentiated areas consisting of a highly cellular, primitive mesenchymal component with numerous mitotic figures and extensive anaplasia to well-differentiated portions expressed as scattered foci of mature cartilage consisting of single and multiple chondrocytes isolated within lacunae. The lesions, possibly arising from the sclera, were invasive into the eye, with elements of the choroid and striated muscle trapped within the tumor matrix. Histologically these lesions resemble the paddlefish chondrosarcoma and mesenchymal chondrosarcomas in higher vertebrates.

Courtney, Lee A. 1996. Cholangioma in a Wild-Caught Sheepshead Minnow (Cyprinodon variegatus) from the Northern Gulf of Mexico. Gulf Res. Rep. 9(3):211-213. (ERL,GB 923).

A single case of a cholangioma occurred in the liver of a wild-caught sheepshead minnow (Cyprinodon variegatus). This is the first biliary neoplasm and second case of a hepatic neoplasm reported from a wild-caught specimen of this species. The findings further demonstrate the susceptibility of the sheepshead minnow to neoplasm development and add support to its selection as a subject for field monitoring of carcinogenic exposure.

Courtney, Lee. 1998. Gulf of Mexico Aquatic Mortality Response Network (GMNET) (Internet). EPA/600/R-98-066. U.S. Environmental Protection Agency, National Health and Environmental Effects Research Laboratory, Gulf Breeze, FL.

http://www.gmpo.gov/gmnet/homepage.htm. 35 p. Mortalities of fish, shellfish, plants, birds and marine mammals occur continuously in the Gulf of Mexico as a consequence of natural events or of conditions imposed on the aquatic community by the activities of man. Reporting of unusual mortality events can serve to signal public health dangers and/or degrading environmental conditions. The Gulf of Mexico Aquatic Mortality Network (GMNET) is a product of many scientists and citizens who share a common interest in establishing and maintaining a reliable response network in the Gulf. GMNET is organized into three interacting groups: Mortality Response, Science Expertise, and Data Management.

Mammalian P-glycoprotein is a highly conserved integral membrane protein functioning as an energy-dependent efflux pump which decreases the concentration of certain lipophilic aromatic compounds entering the cell by diffusion. Expression of such a transporter in teleost species could play a significant role in conferring resistance to fish populations exposed to xenobiotic stressors and may serve as a potential indicator of species at risk to certain environmental contaminants. In previous studies we demonstrated that a strong correlation existed between corresponding mammalian and teleost tissues showing immunoreactivity to specific mammalian P-glycoprotein antibodies. In the present study, comparisons of staining pattern, intensity, and tissue specificity between tissues treated in Bouin's Dietrich's and Lillie's histological fixatives were determined in the sheepshead minnow, Cyprinodon variegatus,using monoclonal antibodies C219, C494, JSB-1 and polyclonal antiserum MDR(Ab-1). Immunoreactivity of these antibodies was found to be fixative-dependent. Results are presented illustrating the differential staining patterns and tissue specificity observed for each tissue type, fixative, and antibody combination. Our data indicate tissue fixation has a significant impact on P-glycoprotein antibody immunoreactivity in teleost tissues and must be considered in the comparison and interpretation of results.

Couch, John A., Lee A. Courtney and Steven S. Foss. 1981. Laboratory Evaluation of Marine Fishes as Carcinogen Assay Subjects. In: Phyletic Approaches to Cancer. EPA-600/D-80-039. Clyde J. Dawe, Editor. Japan Scientific Society Press, Tokyo. Pp. 125-139. (ERL,GB 171). (Avail. from NTIS, Springfield, VA: PB82-205824)

The U.S. Environmental Protection Agency (EPA) and the National Cancer Inst. (NCI) have major responsibilities for determining the fate and risks of carcinogenic agents in the natural environment. Under the auspices of EPA/NCI, the Carcinogen Research Team at the U.S. EPA Lab, Gulf Breeze, has a major role in investigating the fate, effects, and risks of carcinogenic agents in the aquatic portion of the biosphere. In regard to this role, there is a need for practical, experimental exposure systems for the short term, and long term exposure of fishes and invertebrates in order to evaluate their responses to environmentally significant carcinogens. We have designed and tested an adaptable, aquatic laboratory system for flowing water or static water assays of carcinogenic or suspect carcinogenic agents against marine fishes. We report here the design, results of long term tests, and the future uses of the system for determining the risks of carcinogenic agents in the aquatic environment, and as a system complementary to mammalian assay systems, but which permits the phylogenetic expansion of carcinogen assay methodology. A pilot test of the described system has been completed. This test utilized flowing filtered, estuarine water, controlled water temperature, controlled photo period, controlled nutrition of test species, oxygen concentration monitoring, and various life cycle stages of the test fish, the sheepshead minnow, Cyprinodon variegatus, and the suspect carcinogenic herbicide, trifluralin. Continuous exposures to 1 to 5 µg/l trifluralin were conducted with zygote, thru embryogenesis to adult stages of the fish.

Curtis, Lawrence R., Michael J. Hemmer and Lee A. Courtney. 2000. Dieldrin Induces Cytosolic 7,12-[3H-]Dimethylbenz[a]anthracene Binding but not Multidrug Resistance Proteins in Rainbow Trout Liver. EPA/600/J-00/446. J. Toxicol. Environ. Health Part A. 60(4):275-289. (ERL,GB 1082).

Previously it was demonstrated that biliary excretion of a single dose of [14 C]dieldrin or [3H]7,12-dimethylbenz[a]anthracene (DMBA) was stimulated up to 700% and 300%, respectively, in rainbow trout fed 0.3-0.4 mg dieldrin/kg/d for 9-12 wk. This was not explained by increased activities of hepatic microsomal xenobiotic-metabolizing enzymes or increased amounts of any of six cytochrome P-450 isozymes quantitated by Western blots. It was hypothesized that stimulated excretion was explained by induction of (1) cytosolic binding proteins that facilitated intracellular trafficking of DMBA to sites of metabolism, or (2) ATP-dependent proteins that transport xenobiotic metabolites from liver to bile. Binding of 15 and 60 nmol [3H]DMBA/mg protein increased about 200% in hepatic cytosol from dieldrin-fed fish. A 50-fold molar excess of unlabelled DMBA reduced binding of 15 nmol [3H]DMBA/mg protein (non-specific binding) by the same amount in cytosol from control and dieldrin-fed fish, indicating that dieldrin induced specific binding. Liver sections from control and dieldrin-fed fish were treated with multidrup resistance (MDR) protein monoclonal antibodies C494, C219 and JSB-1, and polyclonal antibody MDR Ab-1. There were no marked differences in optical densities of immunohistochemical staining near bile canaliculi of control and dieldrin-fed fish. Induction of xenobiotic binding capacity in cytosol of dieldrin-fed rainbow trout at least partially explained altered DMBA disposition in fish pretreated with this cyclodiene insecticide.

Fournie, John W., J. Kevin Summers, Lee A. Courtney, Virginia D. Engle and Vicki S. Blazer. 2001. Utility of Splenic Macrophage Aggregates as an Indicator of Fish Exposure to Degraded Environments. J. Aquat. Anim. Health. 13(2):105-116. (ERL,GB 1105).

The utility of splenic macrophage aggregates (MAs) as an indicator of fish exposure to degraded environments was evaluated in several species of estuarine fishes as part of the Environmental Protection Agency's Environmental Monitoring and Assessment Program - Estuaries (EMAP-E). Using image analysis, the number and mean size of MAs/mm2 were measured on tissue sections of spleen from a total of 983 fishes representing 7 species from 266 stations scattered across the Gulf of Mexico coastal estuaries. These samples were analyzed for MAs and at 16 stations at least one fish exhibited high densities (>40 MAs/mm2 ). Densities of MAs that exceeded 40/mm2 correlated with exposure to either hypoxic conditions or sediment contamination. Fisher's exact test showed observed frequencies of joint occurrence between high numbers of MAs and both high sediment contaminants and low dissolved oxygen were higher than the expected background frequencies. For all 16 sites where MAs >40/mm2, sediments displayed at least one contaminant at a concentration in the highest 5% of those observed for all Gulf of Mexico stations. This study demonstrates that splenic MAs are effective biotic indicators that can be used to discriminate between fish exposed to degraded and non-degraded environments.

Winstead, James T. and Lee A. Courtney. 2003. Ovacystis-like Condition in the Eastern Oyster Crassostrea virginica from the Northeastern Gulf of Mexico. EPA/600/J-03/147. Dis. Aquat. Org. 53(1):89-90. (ERL,GB 1167).

Histological examination of the eastern oyster, Crassostrea virginica, from a study in Pensacola Bay, Florida, revealed 2 cases of abnormally large, basophilic ova that resembled cells characteristic of ovacystis disease previously reported in oysters from Maine and Long Island. The hypertrophied gametes measured up to 250 µm in diameter, had scant cytoplasm and contained granular nuclear-masses of Feulgen-positive material. Electron microscopy of reclaimed tissue revealed these masses to consist of virus-like particles (average 46 nm) similar to those reported in cases of ovacystis.

Fisher, William S., Deborah L. Santavy, William P. Davis and Lee A. Courtney. 2006. Regional Monitoring of Coral Condition in the Florida Keys. In: USDA Forest Service Proceedings RMRS-P-42CD. 2006. Proceedings of the Monitoring Science and Technology Symposium: Unifying Knowledge for Sustainability in the Western Hemisphere, held 20-24 September 2004, Denver, CO. Aguirre-Bravo, Celedonio, et al., Editor. U.S. Department of Agriculture, Forest Service, Rocky Mountain Research Station, Fort Collins, CO. Pp. 304-311. (ERL,GB 1219).

Tropical reef corals have experienced unprecedented levels of bleaching and disease during the last three decades. Declining health has been attributed to several stressors, including exposures to elevated water temperature, increased solar radiation, and degraded water quality. Consequences of coral bleaching and disease vary; some recover, while others lose tissue, die and succumb to algal overgrowth. In 2000, a regional monitoring project documented disease prevalence and bleaching across 41 km2 of coral reefs in the Florida Keys. Thirty sites were randomly selected from a spatially-balanced grid. A radial belt transect (113 m2) was surveyed at each site and 100-300 colonies were encountered in each transect. The coral species and health status were recorded for each colony. No site had greater than 13% disease prevalence and approximately 80% of the reef area had <5% disease prevalence. The survey will be repeated in 2005, but with additional measurements to estimate colony size, percent living tissue, and living surface area. These added endpoints are expected to provide information on the consequences of bleaching, disease, and other stressors on coral communities. Data will be compiled to characterize community composition, abundance, age class structure, and survival of different species across the Florida Keys reef tract. During a pilot study in 2003, living coral tissue on large colonies of elkhorn coral Acropora palmata was considerably less than on small colonies, possibly indicating a major mortality event (e.g., hurricane or bleaching) that occurred prior to recruitment of the smaller colonies.

Fournie, John W., Marilyn J. Wolfe, Jeffrey C. Wolf, Lee A. Courtney, Rodney D. Johnson and William E. Hawkins. 2005. Diagnostic Criteria for Proliferative Thyroid Lesions in Bony Fishes. Toxicol. Pathol. 33(5):540-551. (ERL,GB 1224).

Thyroid proliferative lesions are rather common in bony fishes but disagreement exists in the fish pathology community concerning diagnostic criteria for hyperplastic versus neoplastic lesions. To simplify the diagnosis of proliferative thyroid lesions and to reduce confusion regarding lesion interpretation, we propose specific criteria for distinguishing hyperplastic from neoplastic lesions. Development of these criteria was based on the examination of a large series of proliferative lesions from Japanese medaka (Oryzias latipes), lesions from other small fish species, and a reexamination of the 97 cases of proliferative thyroid lesions from bony fishes deposited in the Registry of Tumors in Lower Animals. Specific diagnostic criteria are provided for all lesion categories including follicular cell hyperplasia (simple, nodular, or ectopic), adenoma (papillary or solid), and carcinoma (well- or poorly differentiated).These criteria should assist fish pathologists in describing and categorizing naturally-occurring proliferative lesions from wild fishes, lesions that develop in laboratory fishes due to suboptimal culture practices or water quality, those in fishes used in toxicological assays, and captive aquarium fishes.

Barron, Mace G., Cheryl J. McGill, Lee A. Courtney and Dragoslav T. Marcovich. 2010. Experimental Bleaching of a Reef-Building Coral Using a Simplified Recirculating Laboratory Exposure System. J. Mar. Biol. 2010(415167):8p. (ERL,GB 1341).

Determining stressor-response relationships in reef building corals is a critical need for researchers because of global declines in coral reef ecosystems. A simplified recirculating coral exposure system for laboratory testing of a diversity of species and morphologies of reef building corals under ecologically relevant conditions of temperature, salinity, and solar radiation is described. The system provides reproducible control of environmental parameters including temperature, salinity, and solar radiation. Experiments using the model reef building coral, Pocillopora damicornis, show temperature and solar radiation dependent changes in pigment, numbers of symbiotic algae, photosystem II yield, and tissue loss during exposure and recovery. The laboratory exposure system is adaptable for use in experimental bleaching with multiple species of reef building corals

Couch, John A., Lee A. Courtney, James T. Winstead and Steven S. Foss. 1979. American Oyster (Crassostrea virginica) as an Indicator of Carcinogens in the Aquatic Environment. In: Animals as Monitors of Environmental Pollutants. EPA-600/J-79-080. National Academy of Sciences, Washington, DC. Pp. 65-84. (ERL,GB 338). (Avail. from NTIS, Springfield, VA: PB80-18524)

The American oyster (C. virginica) was used as the experimental animal for chronic exposure to 3-methylcholanthrene (3-MC) and benzo[a]pyrene (BP) in an exposure system in which the carcinogens can be continuously injected into free flowing water at fixed rates ranging from 1 to 5 µg/l. Experiments designed to determine uptake and distribution of H3MC and H3BP showed that these are concentrated in oyster tissues in direct proportion to the dosage of carcinogen injected into the system. Residual concentrations as high as 84.4 µg/kg of MC and 36.4 µg/kg of BP were present in oysters as long as 6 months following exposure. Autoradiography showed intense localization of H3BP in distal portions of the tubules of the digestive gland and to a lesser extent in the gonadal tissues. Aryl hydrocarbon hydroxylase (AHH) activity was present in homogenates of hepatopancreas after 5.5 months of exposure to the carcinogens, in contrast to control animals in which AHH activity was quite low. In eight oysters exposed to MC, an infiltration of cells believed to be of hematopoietic origin was encountered in the mantle. Some appear to be identical in type to those which constitute sarcoma-like lesions encountered in feral oyster populations. However, it would be premature at this stage to assign any etiological significance to the experimental findings.

Couch, John A. and Lee A. Courtney. 1985. Attempts to Abbreviate Time to Endpoint in Fish Hepatocarcinogenesis Assays. In: Water Chlorination: Chemistry, Environmental Impact and Health Effects, Vol. 5. EPA-600/D-84-229. Robert L. Jolley, Editor. Lewis Publishers, Chelsea, MI. Pp. 377-398. (ERL,GB 518). (Avail. from NTIS, Springfield, VA: PB84-246297)

In recent years, the use of freshwater and marine fishes in carcinogen research and in environmental carcinogen monitoring has grown substantially. Several advances must be made with selected species to make fishes advantageous and practical as assay subjects. Some of these advances should be (1) precise characterization of neoplastic endpoints and progression in experimentally exposed fishes, (2) abbreviation of length in time needed for risk evaluation of carcinogens or suspect agents in fishes, and (3) correlation of endpoints for carcinogen effects in fishes with those in other more-routine test species such as rodents (mammals). Because we believe that fishes, as a phyletic group, have much to teach us about neoplasia and environmental carcinogenesis, we are studying the experimental induction, progression, and fate of neoplasms in the liver of a marine coastal fish, the sheepshead minnow (Cyprinodon variegatus). The agent used to induce liver lesions in these studies was N-nitrosodiethylamine (DEN). The sheepshead minnow has been used for several years as a toxicological and carcinogen assay subject in our laboratories.5,6 The objectives of this study were to (1) characterize liver neoplastic development and (2) reduce or abbreviate times to endpoints in liver carcinogen assays using the sheepshead minnow with histological, ultrastructural, and enzyme histochemical endpoints.

Fournie, John W., Steven S. Foss, Lee A. Courtney and Albert H. Undeen. 1990. Testing of Insect Microsporidians (Microspora:Nosematidae) in Nontarget Aquatic Species. EPA/600/J-90/376. Dis. Aquat. Org. 8:137-144. (ERL,GB 680). (Avail. from NTIS, Springfield, VA: PB91-163899)

This paper reports results of tests with the mosquito microsporidian Nosema algerae and the orthopteran microsporidian N. locustae on nontarget aquatic organisms. Organisms tested were the freshwater grass shrimp Palaemonetes kadiakensis, the estuarine grass shrimp P. pugio, the marine rotifer Brachionus plicatilis, and the inland silverside Menidia beryllina. These organisms were exposed by intrahemocoelic injection, gavage, or ingestion. Infections did not develop in either the freshwater grass shrimp or the estuarine grass shrimp that were gavaged with N. algerae spores. However, infections did develop in both species of grass shrimp after intrahemocoelic injections with N. algerae spores. Infected tissues included the gills, antennal gland, eyes, skeletal muscle, heart, and gonads. Proof of infection was demonstrated ultrastructurally by the presence of mature spores and developmental stages in infected tissues. Infections did not develop in P. pugio following intrahemocoelic injections of N. locustae spores. N. algerae infections did not develop in P. pugio following intrahemocoelic injections of N. locustae spores. N. algerae infections did not develop in the marine rotifer after ingestion of spores or in inland silversides fed marine rotifers containing ingested spores.

Winstead, James T., Douglas P. Middaugh and Lee A. Courtney. 1991. Ovarian Mycosis in the Topsmelt, Atherinops affinis (Ayres). EPA/600/J-91/332. Dis. Aquat. Org. 10:221-223. (ERL,GB 702). (Avail. from NTIS, Springfield, VA: PB92-129691)

An ovarian mycosis in a topsmelt, Atherinops affinis (Ayres) collected from Elkhorn Slough, Monterey County, California, is reported. The fungus caused a considerable host response resulting in numerous granulomas, fibrosis and egg compression. Based on morphologic characteristics and the elimination of other fungal pathogens, the fungus is probably a member of the class Oomycetes. This is the first report of an invasive ovarian mycosis in a marine fish.

Diamant, Ariel, John W. Fournie and Lee A. Courtney. 1994. X-Cell Pseudotumors in a Hardhead Catfish, Arius felis (Ariidae), from Lake Pontchartrain, Louisiana, U.S.A.. EPA/600/J-94/340. Dis. Aquat. Org. 18:181-185. (ERL,GB 856). (Avail. from NTIS, Springfield, VA: PB94-198165)

X-cell epidermal lesions are described from a single specimen of the hardhead catfish Arius felis (Aridae). The lesions exhibited an unusual growth pattern but did not involve any visceral organs. Histologically, the lesions resembled those previously described for coldwater fishes. This is the first report of X-cell lesions from Lake Pontchartrain, Louisiana, USA, and the hardhead catfish represents the first warmwater species affected with the disease.

Hemmer, Michael J., Lee A. Courtney and Lisa S. Ortego. 1995. Immunohistochemical Detection of P-glycoprotein in Teleost Tissues Using Mammalian Polyclonal and Monoclonal Antibodies. EPA/600/J-95/372. J. Exp. Zool. 272(1):69-77. (ERL,GB 899).

Mammalian P-glycoprotein is a highly conserved 170-kD integral plasma membrane protein functioning as an energy dependent efflux pump of exogenous and endogenous lipophilic aromatic compounds entering the cell by diffusion. In this study, the tissue specificity of one polyclonal (pAb) and three monoclonal (mAbs) antibodies to mammalian P-glycoprotein were identified in paraffin-embedded, parasagittal whole body sections of the guppy, Poecilia reticulata. Pab mdr (Ab-1) and mAbs C219, C494 and JSB-1 demonstrated differential staining patterns in the following tissues: bile canaliculi in the liver, exocrine pancreas, lumenal surface of the intestinal epithelium, renal tubules, interrenal tissue, branchial blood vessels, gas gland, pseudobranch, and the gill transverse septa. Positive P-glycoprotein expression in P. reticulata correlates well with published results for homogogous mammalian tissues of secretory and execretory function. These data indicate that one or more highly conserved members of the P-glycoprotein transporter family exist in a teleost species which can be detected using commercially available mammalian antibodies.

Glas, Patricia S., Lee A. Courtney, James R. Rayburn and William S. Fisher. 1997. Embryonic Coat of the Grass Shrimp Palaemonetes pugio. Biol. Bull. 192(2):231-242. (ERL,GB 979).

The embryo of the grass shrimp, Palaemonetes pugio, is surrounded during development by a protective extracellular coat designated as the embryonic coat (EC). At hatching, this EC is composed of four embryonic envelopes (EE) each of which is composed of multiple layers. The outermost layer of the EC, the outer investment coat (OIC), is derived primarily, if not completely, from pleopods of the female. The first envelope (EE1) is bilayered and forms immediately after oviposition. The OIC becomes closely associated with EE1 and remains in close contact to EE1 until hatching occurs. An additional layer is added to the inner side of EE1 between 3-5 d after oviposition. Three more EE, EE2, EE3 and EE4, are formed between the embryo and EE1 by 7 d after oviposition. Formation of EE continues until 10 d after oviposition by which time each envelope is morphologically distinct in composition with 'outer' and 'inner' sides clearly identifiable. All but the innermost EE, EE4, is shed by the embryo within 6 h of hatching. The OIC labels positively with fluorescently labeled lectins for mannose and N-acetylglucosamine, but EE4 does not label strongly with any of the lectins used. Permeability of the EC during the 12 d incubation period is found to decrease between 0 and 5 d after oviposition, and then increase until hatching. Thus, the palaemonid EC is a dynamic structure throughout embryonic development.

Hinton, David E., John A. Couch, Swee J. Teh and Lee A. Courtney. 1988. Cytological Changes During Progression of Neoplasia in Selected Fish Species. Aquat. Toxicol. 11(1,2):77-112. (ERL,GB X539).

Cytological changes during progression of hepatic neoplasia in fishes were reviewed with emphasis on recent findings in Cyprinodon variegatus and Oryzias latipes. Hepatocytes are particularly sensitive to toxic changes during early phases of response to carcinogens reflecting both lethal and sublethal alterations. Enzyme histochemical studies reveal marked deficiency of glucose-6-phosphate dehydrogenase, glucose-6-phosphatase and adenosine triphosphatase. Surviving hepatocytes are either enlarged, encircled by cells with small nuclear to cytoplasmic rations, and have altered nuclear morphology suggestive of an inability to divide, or, are smaller, apparently rapidly dividing, and have basophilic cytoplasm. In both species, development of spongiosis hepatis occurred following cytotoxic phases. This lesion apparently provides abundant space for cellular remodeling during neoplastic progression leading to eventual multinodular change. Foci of altered hepatocytes included basophilic, eosinophilic (both species) and clear cells (Cyprinodon variegatus only). Enzyme alterations preceded other morphologic alterations and were seen in cells of foci and tumors suggesting lineage of phenotypic alteration. Cytologic changes within other resident cell populations during neoplastic progression were reviewed.