Effects of two oils and 16 pure polycyclic aromatic hydrocarbons on plasmatic immune parameters in the European sea bass, Dicentrarchus labrax (Linné).

The in vitro effects of polycyclic aromatic hydrocarbons (PAHs) on two plasmatic immune parameters, lysozyme concentration and haemolytic alternative complement activity, of the European sea bass, Dicentrarchus labrax, were tested using field (10(-7) and 10(-9) mg mL(-1)) and high concentrations (10(-3) and 10(-5) mg mL(-1)) observed during oil spills. Peripheral blood from 105 fish was collected, centrifuged at 1200 g, for 10 min, at 4 degrees C and three plasma pools, each of 35 fish, were constituted. Two oils (heavy fuel oil and light cycle oil) and 16 pure PAHs, selected on the basis of the American Environmental Protection Agency list (US EPA), were tested in vitro on the two humoral immune parameters. Only three pure PAHs (anthracene, chrysene and dibenz[a,h]anthracene) modulated lysozyme concentration. Acenaphthene, acenaphthylene, anthracene, benzo[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, pyrene and light cycle oil modified the haemolytic alternative complement activity after 4h of incubation. This study investigates the direct effects of several PAHs on fish humoral immune functions and describes the haemolytic complement activity of fish as suitable biomarkers of oil pollution.

Study of the viral interference between infectious pancreatic necrosis virus (IPNV) and infectious haematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss).

The resistance of rainbow trout (Oncorhynchus mykiss) to an infectious haematopoietic necrosis virus (IHNV) challenge following a preceding non-lethal infection with infectious pancreatic necrosis virus (IPNV) was investigated through experimental dual infections. Trout initially infected with IPNV were inoculated 14 days later with IHNV. Single infections of trout with 1 of the 2 viruses or with cell culture supernatant were also carried out and constituted control groups. No mortality was noted in fish after a single infection with IPNV. This virus had no influence on the head kidney leucocyte phagocytic activity and plasma haemolytic complement activity. IHNV induced a high mortality (72%) and reduced the macrophage phagocytic activity and complement haemolytic activity. It also induced a late production of anti-IHNV antibodies which occurred after clearance of the virus in the fish. In trout co-infected with both viruses, a mortality rate of 2% occurred and the immune parameters were similar to those observed in the fish infected with IPNV only, demonstrating that in co-infected trout IPNV inhibits the effects of IHNV. The studied parameters did not allow us to define the mechanism of interference occurring between these 2 viruses, but some hypothesis are put forward to explain the interference between the 2 viruses.

Pathogenesis and immune response in Atlantic salmon (Salmo salar L.) parr experimentally infected with salmon pancreas disease virus (SPDV).

Atlantic salmon parr were injected intraperitoneally with salmon pancreas disease virus (SPDV) grown on CHSE-214 cells. The viraemia, the histopathological changes in target organs and some immune parameters were taken at intervals up to 30 days post-infection (dpi). The earliest kind of lesion was necrosis of exocrine pancreas, appearing as soon as 2 dpi. It progressed towards complete tissue breakdown at 9 dpi before resolving gradually. Concurrent to this necrosis, a strong inflammatory response was in evidence from 9 dpi in the pancreatic area for a majority of fish. A necrosis of the myocardial cells of the ventricle occurred in infected fish mainly at 16 dpi and it faded thereafter. The monitoring of the plasma viral load showed a rapid haematogenous spreading of SPDV, peaking at 4 dpi, but also the absence of a secondary viraemia. No interferon (IFN) was detected following the infection of parr with SPDV, probably owing to an IFN activity in Atlantic salmon below the detection level of the technique. Neutralising antibodies against SPDV were in evidence from 16 dpi and they showed a time-related increasing titre and prevalence. The phagocytic activity in head-kidney leucocytes was always significantly higher in the infected fish than in the control fish, being particularly high by 9 dpi. Lysozyme and complement levels were both increased and they peaked significantly in the infected fish at 9 and 16 dpi respectively. These results demonstrated that an experimental infection of Atlantic salmon parr with SPDV provoked a stimulation of both specific and non-specific immunity with regards to the viraemia and the histopathology.

Immunocytochemical detection of Ig-positive cells in blood, lymphoid organs and the gut associated lymphoid tissue of the turbot (Scophthalmus maximus).

The present study was designed to search for the sites of the B-cell lineage in the different lymphoid organs of turbot (Scophthalmus maximus) by immunoperoxidase staining with a rabbit polyclonal antiserum against deglycosylated turbot IgM (TUDG-6). A turbot immunoglobulin (Ig) fraction, isolated by protein A, was checked for purity by gel filtration and SDS-PAGE under reducing conditions. The turbot IgM was deglycosylated and used to raise an antiserum. The antiserum titre was evaluated in ELISA. It was then used to analyse turbot peripheral blood leucocytes for membrane and cytoplasmic Ig and for immunohistochemistry with turbot lymphoid tissues. Very low numbers of Ig+ cells were found in thymus sections. In sections of spleen, Ig+ cells were observed in white pulp, around ellipsoids but were mostly concentrated and associated with melanomacrophage centers (MMCs). The lymphoid Ig+ cells in the kidney tended to be dispersed among haematopoietic and granulopoietic cell populations and were in intimate association with the MMCs and blood vessels. This association between MMCs and Ig+ cells in the spleen and the kidney, is discussed with respect to the role played by these organs in the immune system of fish. Last, the lymphoid population in the gut associated lymphoid tissue (GALT) of turbot was characterised with respect to staining for Ig. Immunoreactive cells were rarely detected in the epithelial layer although many lymphocytes were present, but they were frequently observed in the lamina propria, presumably as part of the GALT and involved in mucosal immune responses.

Antigen uptake and immune responses after oral vaccination.

Oral delivery of vaccine, with antigen simply mixed with feed, is the easiest method of mass vaccination of fish of all sizes. It is time and labour-saving, and avoids any manipulation stress. However, large quantities of antigen are necessary and it is not possible to know the exact dose absorbed by each fish. Moreover the resulting protection is generally weak and of short duration. To improve the effectiveness of oral vaccination a considerable amount of work has been done in the last 15 years. It is now well established that the second segment of the hindgut of fish has the ability to absorb soluble or particulate antigens. These antigens reach the intraepithelial macrophages which show antigenic determinants on their cell membrane, suggesting an antigen-presenting function. Moreover, a gut-associated lymphoid tissue composed of several lymphoid cell types, macrophages, granulocytes and plasma cells has been described in this region of the gut and recent studies indicate that it is involved in an intestinal immune response. Both mucosal and systemic immune responses seem to develop as indicated by the presence of antibodies in gut mucus, bile and serum. However, these responses are higher following anal delivery of antigen than oral delivery. It is generally considered that during oral delivery antigens are digested in the foregut and/or in the stomach. Thus studies are being carried out now on ways to protect antigens, for instance through the encapsulation of antigen, the neutralisation of gastric secretions and the use of oral adjuvants.

Enzootic bovine leukosis: an epidemiological survey among cattle in the west of France by precipitating antibody detection.

In order to inform the sanitary veterinaty authorities, an epidemiological survey was carried out in the West of France to appreciate the level of infection by BLV in cattle. More than 10,000 sera from 763 herds were tested by agar gel immuno-diffusion. The animals were more than one-year-old dairy cattle. 23 sera (0.22%) from 11 herds (1.44%) were found positive. In the 11 herds the levels of serologically positive animals ranged from 1.6 to 40% with a 9% average. These levels are comparatively low and are almost in correspondence with the number of leukotic tumours recorded in the slaughter houses and knackeries.