bromochloroacetic-acid and Fish-Diseases

A cell line, CTE, derived from catla (Catla catla) thymus has been established by explant method and subcultured for more than 70 passages over a period of 400 days. The cell line has been maintained in L-15 (Leibovitz) medium supplemented with 10% fetal bovine serum. CTE cell line consists of homogeneous population of epithelial-like cells and grows optimally at 28°C. Karyotype analysis revealed that the modal chromosome number of CTE cells was 50. Partial amplification, sequencing and alignment of fragments of two mitochondrial genes 16S rRNA and COI confirmed that CTE cell line originated from catla. Significant green fluorescent signals were observed when the cell line was transfected with phrGFP II-N mammalian expression vector, indicating its potential utility for transgenic and genetic manipulation studies. The CTE cells showed strong positivity for cytokeratin, indicating that cell line was epithelial in nature. The flow cytometric analysis of cell line revealed a higher number of cells in S-phase at 48 h, suggesting a high growth rate. The extracellular products of Vibrio cholerae MTCC 3904 were toxic to the CTE cells. This cell line was not susceptible to fish betanodavirus, the causative agent of viral nervous necrosis in a large variety of marine fish.

Topics: Animals; Cattle; Cell Line; Cyprinidae; Epithelial Cells; Fish Diseases; Fish Proteins; Karyotype; Keratins; Nodaviridae; RNA Virus Infections; RNA, Ribosomal, 16S; S Phase; Thymus Gland

An increasing interest in fish species as sentinels of environmental pollution and in carcinogenesis research has led to the identification of diagnostically challenging neoplasms of uncertain cellular origin and the need for additional diagnostic methods. To determine the potential of using commercially available antibodies to intermediate filament proteins on paraffin-embedded fish tissues for immunocytochemistry in tumor diagnosis, the application of three antikeratin antibodies to normal adult tissues from two fish species was assessed. Multiple tissues from 12-14-in. striped bass (Morone saxatilis) and 6-month-old medaka (Oryzias latipes) of both sexes were fixed in Bouin's or formalin fixatives. Formalin-fixed neoplasms from several mammalian species, including cat, dog, hedgehog (Atelerix albiventris, Erinaceus europaeus), rhesus macaque (Macaca mulatta), and sloth bear (Melursus ursinus), were also used as positive controls. Using a strepavidin horseradish peroxidase method on paraffin-embedded tissues, the broad spectrum antibodies AE1/AE3 (Boehringer Mannheim, Indianapolis, IN) and MAK-6 (Triton Biosciences, Alameda, CA), which recognize most of the 19 human cytokeratins, and CAM 5.2 (Becton Dickinson, Mountain View, CA), which recognizes cytokeratins present in human liver, were used as primary antibodies. Epithelia from skin, gills, cornea, bile ducts, renal tubules, gastrointestinal tract, and thymus were strongly positive with AE1/AE3 and MAK-6 in striped bass, but nonepithelial tissues such as bone and muscle were negative. Skin, gills, cornea, and portions of the gastrointestinal tract were strongly positive in medaka with the same antibodies, whereas bile duct, renal, and intestinal epithelia were less so. Tissue digestion improved the intensity of staining, and fixation with Bouin's fixative improved results somewhat compared with formalin fixation.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Bass; Fish Diseases; Immunoenzyme Techniques; Keratins; Neoplasms; Oryzias; Species Specificity

Eggshells of the palaeacanthocephalans Acanthocephalus anguillae, Pomphorhynchus laevis and Polymorphus minutus were investigated for their fine structure as well as their chemical composition. The acanthor larvae are surrounded by four eggshells (E1-E4) separated by interstices (G1-G4). Immature eggs do not exhibit the complete set of eggshells. The chemical composition of the outermost, thin eggshell (E1) remains unknown. E2 is supplied by outer filaments of different strength; it contains keratin, which was localized electron microscopically using anti-keratin. In P. laevis and P. minutus, E3 seems to contain glycoproteins, which could not be visualized in this eggshell of A. anguillae. The innermost eggshell (E4) uniformly contains chitin. The electron-lucent interstices of the eggs of P. laevis and P. minutus are rich in polysaccharides and/or proteoglycans, whereas those of A. anguillae contain low amounts of such substances. The differences observed in the abundance of carbohydrates are discussed with respect to the life cycles of the three acanthocephalans.

Topics: Acanthocephala; Animals; Animals, Wild; Bird Diseases; Cyprinidae; Ducks; Fish Diseases; Helminthiasis; Helminthiasis, Animal; Immunohistochemistry; Keratins; Microscopy, Electron; Ovum; Polysaccharides; Proteoglycans; Species Specificity

A battery of polyclonal and monoclonal antibodies directed against cytokeratins, desmin, vimentin, glial fibrillary acidic protein and neurofilament proteins of mammalian species was used to demonstrate intermediate filament (IF) expression in normal and lymphoma tissues of northern pike by indirect immunofluorescence microscopy. Frozen sections of pike intestine, skin, skeletal muscle, heart, liver, testis, head-, mid-, and posterior kidney and brain demonstrated IF in a manner which strengthens the idea that they are evolutionarily highly conserved. The results also confirm the IF-subclass specificities for different types of tissues, as noted by others in other species. Pike lymphoma cells showed morphological resemblance to head kidney cells; immunofluorescence microscopy and immunoblotting revealed vimentin expression, suggesting that the Aland pike lymphoma is a true mesenchymal neoplasm and is derived from haemic cells. The significance of these studies in relation to similar work with other species and to the possible use of IFs in the classification of normal and diseased tissues in fish is discussed.

Topics: Animals; Cytoskeleton; Desmin; Electrophoresis, Polyacrylamide Gel; Fish Diseases; Fluorescent Antibody Technique; Glial Fibrillary Acidic Protein; Immunologic Tests; Intermediate Filament Proteins; Intermediate Filaments; Keratins; Lymphoma; Microscopy, Fluorescence; Salmonidae; Vimentin