In vivo effect of the organophosphorus insecticide trichlorphon on the immune response of carp (Cyprinus carpio). I. Effect of contamination on antibody production in relation to residue level in organs.

The organophosphorus insecticide trichlorphon was tested at medium (10 and 20 ppm) and high concentrations (20,000 ppm) on antibody production against Yersinia ruckeri vaccine. No statistically significant effect was observed on humoral response or on hematocrit. Contamination of organs was analyzed by acetylcholinesterase activity. At 20,000 ppm only liver and brain were highly contaminated; spleen and lymphocytes were not significantly contaminated. At 20 ppm no residue was found in organs.

In vivo effect of the organophosphorus insecticide trichlorphon on immune response of carp (Cyprinus carpio). II. Effect of high doses of trichlorphon on nonspecific immune response.

The effect of trichlorphon, one of the most widely used organophosphorus insecticides, on the nonspecific immune response in carp (Cyprinus carpio) was studied. The effect of 20,000 ppm trichlorphon on the immune response was followed for 3 and 56 days after intoxication. The effect of 10,000 ppm trichlorphon on the nonspecific immune response of carp experimentally infected by Pseudomonas alcaligenes and Aeromonas punctata was also examined. Leucocyte number, phagocytic ability of neutrophils, percentage NBT-positive PMN cells, phagocytic index, lysozyme level in serum, and ceruloplasmin activity in plasma were examined on Days 2, 4, 6, 8, 10, 14, 18, 22, and 26 after carp were exposed. After intoxication leukopenia was observed as decreases in phagocytic ability of neutrophils and in phagocytic index. Lysozyme level in serum was also decreased compared to that of control. The percentage of NBT-positive PMN cells decreased when the ceruloplasmin activity in plasma increased in intoxicated fish.

Effect of levamisole on the lymphocyte and macrophage activity in carp (Cyprinus carpio).

Levamisole, an anthelminthic drug which is often used in cattle, is known for its immunomodulatory effect in several species. In the present work we investigated the in vitro effects of different concentrations of levamisole on lymphocyte proliferation with mitogen in carp. At low concentrations, ie 12.5 to 0.1 micrograms/ml, levamisole stimulated proliferation with phytohemagglutinin P (PHA-P) and Con A, had no effect at medium concentrations and acted as a suppressor at high concentrations, ie 100 micrograms/ml PHA-P and 100 to 10 micrograms/ml for Con A. Spleen and kidney macrophages were in contact with levamisole for 2 h before phagocytosis of opsonised Yersinia ruckeri. Doses of 200 and 100 micrograms/ml were found to be suppressive, whereas 6.2-0.8 micrograms/ml were stimulatory, as observed by chemiluminescence.

Effect of manganese ions on the immune response of carp (Cyprinus carpio) against Yersinia ruckeri.

The effect of manganese ions Mn2+ (Mn Cl2) on hematocrit and the antibody response of carp (Cyprinus carpio) was investigated after exposure of fish to contaminated water for 2.5 months. Contamination of water with 50 mg/l Mn2+ did not decreased antibody production against Yersinia ruckeri (measured by agglutination) but significantly decreased hematocrit. The levels of manganese in lymphoid organs (spleen and kidney) increased progressively during the experiment. The effect of manganese in vitro on phagocytosis of opsonized Yersinia ruckeri was studied by chemiluminescence. Mn2+ has a stimulating effect on phagocytosis in a range of 6.25 to 50 mg/l.

Effect of pesticides (atrazine and lindane) on the replication of spring viremia of carp virus in vitro.

Atrazine and lindane are widely employed as pesticides in agriculture, which by wash off, could contaminate the aquatic environment. In this study, they were tested in vitro on the replication of the virus of Spring Viremia of Carp (SVC) in Epithelioma Papulosum Cyprini cell line. They were dissolved first in DMSO and then in medium at their solubility limit in water (28 and 10 mg/l). The results of the kinetic titers after 72 h showed no statistically significant difference between control and contaminated cells.

Effect of gamma-hexachlorocyclohexane (lindane) on carp (Cyprinus carpio). I. Effect of chronic intoxication on humoral immunity in relation to tissue pollutant levels.

Lindane, the gamma-isomer of hexachlorocyclohexane, one of the most widely used insecticides, was incorporated into carp food for 109 days. The effects of 10, 100, or 1000 ppm of lindane in food on hematocrits and on antibody production against Yersinia ruckeri bacterin in sera and mucus were followed for 79 days. No effect of lindane on these parameters and on spleen weight was observed. Lymphoid organs, spleen and kidney, were as highly contaminated at the beginning of the inoculation (Day 30) as at the end of the experiment. The tissue levels of lindane in liver, spleen, kidney, or whole body, determinated after 30 and 109 days of contamination, indicated that the tissue distribution was dependent on organ lipid contents.

Effect of gamma-hexachlorocyclohexane (lindane) on carp (Cyprinus carpio). II. Effects of chronic intoxication on blood, liver enzymes, and muscle plasmic membrane.

A 30-day ingestion of gamma-hexachlorocyclohexane (lindane) by carp (Cyprinus carpio) induced hypoglycemia without activation of two hepatic gluconeogenesis enzymes (fructose diphosphatase, EC 4.1.2.13, and glucose-6-phosphatase, EC 3.1.3.9) and hyponatremia and variations in muscle plasmic membrane-bound enzymes (especially cholinesterases, EC 3.1.1.7). After 109 days carps exhibited a decrease in natremia but no significant hypoglycemia. There was an activation of gluconeogenesis enzymes. Important changes were observed in the activities of muscle plasmic membrane enzymes (especially 5'-nucleotidase, EC 3.1.3.5, and ATPases, EC 3.6.1.3). Lindane, a lipophilic substance, especially disturbed the activity of membrane-bound enzymes enclosed in a phospholipid matrix.

Antibody diversity in trouts obtained by gynogenesis or self-fertilization. Comparative analysis of the heavy chain spectrotypes.

Conventionnal (C) trouts and trouts obtained by gynogenesis (G) or self-fertilization (SF) were immunized with DNP-KLH and anti-DNP antibodies were individually purified by affinity chromatography. The isolated IgM-like antibodies were separated by an iso-electrofocusing technique in reducing conditions and electroblotted onto nitrocellulose. The transfers were probed with a mouse monoclonal antibody specific for trout heavy (H) antibody chain and revealed with a rabbit anti-mouse IgG horse-radish peroxydase conjugate. When comparing the IEF H chain spectrotypes of C, G and SF trouts, it was observed that individual C spectrotypes are more different between themselves than G and SF spectrotypes, and that individual SF spectrotypes were less heterogeneous than C or G ones. These results suggest that in trout, inbreeding induces a reduction of antibody diversity and heterogeneity. The inheritance of antibody repertoire might be taken in account in the inbreeding selection schedules for fish of economical interest.

Variability in humoral responses to DNP-KLH of rainbow trout (Salmo gairdneri). Comparison of antibody kinetics and immunoglobulins spectrotypes between normal trouts and trouts obtained by gynogenesis or self-fertilization.

The anti-DNP antibody responses were compared between conventional trouts produced in fishfarm and trouts experimentally obtained by self-fertilization of hermaphrodites or by gynogenesis. The variability of anti-DNP antibody titres was equivalent between normal and gynogenetic fish, but significantly reduced for fish obtained by self-fertilization. Isoelectrofocusing analysis of individually-isolated anti-DNP antibodies indicate low molecular heterogeneity but large variability between conventional trouts. Almost complete sharing of individual spectrotypes was observed between self-fertilized fish and large similarities between gynogenetics animals. These results suggest that the intensity of antibody synthesis in trout is under a complex genetic control, even in inbred animals expressing large similarities in their antibody repertoires.