Fish Cell Cultures Research Articles

Fish Cell Lines: Comparisons of CHSE-214, FHM, and RTG-2 in Assaying IHN and IPN Viruses

The rainbow trout, Salmo gairdneri, gonad (RTG-2) cell line was more sensitive to lytic infection by infectious pancreatic necrosis virus (IPNV) than was the fathead minnow, Pimephales promelas, (FHM) cell line in laboratory and field studies. Laboratory investigations with known strains of IPNV (VR 299 and Buhl) also indicated that the cytopathic response in IPNV-infected chinook salmon, Oncorhynchus tshawytscha, embryo (CHSE-214) cells was greater than in infected FHM cells. In contrast, titration experiments with standard infectious hematopoietic necrosis virus (IHNV) isolates (Oregon and Cedar River) showed that the FHM cells were more sensitive to destructive infection by IHNV than was either of the salmonid cell lines (RTG-2, CHSE-214). Key words: fish cell culture, infectious pancreatic necrosis virus, infectious hematopoietic necrosis virus, chinook salmon embryo cells, fathead minnow cells, rainbow trout cells, salmonid viruses

Fish Cell Culture: Properties of a Cell Line from the Sheepshead, Archosargus probatocephalus

A cell line designated SHF-1 was established from fin tissue of the sheepshead, Archosargus probatocephalus. Cells were fibroblastlike and grew best at 26 °C in Leibovitz (L-15) medium containing serum supplements and a final sodium chloride concentration of 0.150 M. The 2N number of primary sheepshead cells was 48 but the SHF-1 line at passage levels above 11 was heteroploid. SHF-1 cells did not replicate any of seven viruses tested. Species confirmation of SHF-1 was accomplished by the cytotoxic antibody test and was attempted with isoelectric focusing of lactic dehydrogenase isoenzymes. Plating efficiency and freeze viability studies were also performed. Key words: fish cell culture, sheepshead, Archosargus probatocephalus, microbiology

Fish cell culture: Characteristics of a cell line from the silver perch,Bairdiella chrysura

A cell designated SP-1 was established from tissue of the silver perch, Bairdiella chrysura. Cells were fibroblast-like and grew best at 26 degrees C in Leibovitz medium (L-15) containing 15% fetal bovine serum and 0.150 M sodium chloride. Passage 1 to passage 9 SP-1 cells contained a chromosome number of 48; at passages 27 and 50 the modal numbers were 51 and 54, respectively. Confirmation of the origin of SP-1 cells was made by the cytotoxic antibody dye-exclusion test. This cell line supported the growth of lymphocystis virus from the silver perch but was not found to replicate various other fish and mammalian viruses.

Growth response of the green algae Chlorella vulgaris to selective concentrations of zinc

A modification of a previously published fish cell culture technique (Rachlin and Perlmutter, 1968) has been successfully employed in this study to evaluate the effects of a metal toxicant on the growth response and therefore productivity of the green algae Chlorella vulgaris. The results of this study indicate that the concentration of zinc which reduces the growth rate of a test population of this alga, during a 96 h exposure, by 50 per cent, is 2.4±0.02 ppm. The test design has advantages in that it concerns itself with productivity reduction rather than standard survival (LC 50) of the test organisms. In addition the data generated is readily treated by the system of probit analysis for ease of evaluating the results. The use of this technique and test system, allows for the design of parallel studies in which the effects of toxicants on both animal and plant cell systems can be evaluated in terms of a more unified cellular approach.

Channel catfish virus: a new herpesvirus of ictalurid fish.

Channel catfish virus was studied in ictalurid fish cell culture, the only system of fish, amphibian, avian, and mammalian cells found to be susceptible. Channel catfish virus infection resulted in intranuclear inclusions and extensive syncytium formation. Replication occurred from 10 to 33 C, but not higher. Best growth was from 25 to 33 C, and the amount of virus released nearly equalled the amount which remained cell-associated. The virus was labile to lipid solvents, and indirect determinations with labeled precursors and a metabolic inhibitor showed evidence of deoxyribonucleic acid. Electron microscopy showed progeny virus, about 100 nm in diameter, in various stages of development in cell nuclei by 4 hr. Present also were nuclear masses of exceptionally electron-dense lamellar material, with a unit dimension of 10 to 15 nm. Virus was enveloped at the nuclear membrane and in cytoplasmic vacuoles, resulting in virions having a diameter of 175 to 200 nm. Negative staining demonstrated icosehedral symmetry and 162 capsomeres. Our data indicate that channel catfish virus is a herpesvirus.

Infectious Pancreatic Necrosis: Selection of Virus-Free Stock from a Population of Carrier Trout

Infectious pancreatic necrosis (IPN) is a virulent disease of young trouts and is easily transmitted from infected animals through water and with eggs. At present, the most effective control measure consists of propagation of specific pathogen-free stock. Methods are described for using fish cell cultures to detect IPN virus in peritoneal washes, feces, and gonadal fluids and thereby to identifiy carrier and non-carrier fish. The procedures were used to derive virus-free progeny from a hatchery trout population containing about one-third carrier fish. The progeny were reared to maturity, spawned for 3 successive years, and at each spawning they proved free of virus. Recommendations and precautions are given for those who may wish to apply the procedures.

Lymphocystis virus: isolation and propagation in centrarchid fish cell lines.

A virus from fish with lymphocystis disease was isolated in fish cell cultures. Eleven serial transfers were made and the pathognomonic lymphocystis cells were produced in vitro in each transfer. Fish inoculated with 6th- and 9th-passage material developed the disease, and virus was reisolated front them.

Some Recent Developments and Applications of Fish Cell and Tissue Culture

(1965). Some Recent Developments and Applications of Fish Cell and Tissue Culture. The Progressive Fish-Culturist: Vol. 27, No. 2, pp. 67-74.