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Juvenile Atlantic halibut (~100 mg, Hippoglossus hippoglossus) were exposed to Vibrio proteolyticus, a Vibrio spp. isolate, Photobacterium damselae ssp. damselae and five different isolates of Aeromonas salmonicida ssp. achromogenes via an hour-long bath immersion to ascertain their variation in pathogenicity to this fish species. Results were analysed using Kaplan-Meier survival analysis. Analysis of the data from challenges using A. salmonicida ssp. achromogenes revealed three survival values of zero and a spread of values from 0 to 28.43. Challenges using a Vibrio spp isolate, V. proteolyticus and P. damselae resulted in Kaplan-Meier survival estimates of 31.21, 50.41 and 57.21, respectively. As all bacterial species tested could induce juvenile halibut mortalities, they must all be considered as potential pathogens. However, the degree of pathogenicity of A. salmonicida is isolate dependent. [ABSTRACT FROM AUTHOR]

The ontogenic development of neurosensory structures in larval Atlantic halibut, Hippoglossus hippoglossus (L.) was examined using microscopy imaging techniques and analysis. Larval samples were preserved at developmental stages of 444, 486, 528, 570, 612, and 666 degree-days, respectively. Larvae were measured and photographed macroscopically. The fish were then embedded in paraffin for histological processing. 5 μm sequential sections of the anterior of the larvae were cut and stained using hematoxylin and eosin dyes. The resulting sections were examined using an Olympus BX60 compound microscope. The development of neurosensory structures such as neuromasts and sensory papillae were observed throughout all developmental stages of larvae and early development of the lateral line was observed at 666 degree-days. Neuromast frequency increased as larvae matured, suggesting that larvae were approaching their first feeding event towards the end of the sampling period. Possible implications of development on larval first feeding are discussed. [ABSTRACT FROM AUTHOR]

A fluorescent in situ hybridization (FISH) method was developed for detection of infectious pancreatic necrosis virus (IPNV) in paraffin-embedded tissues of Atlantic salmon, Salmo salar L. Several methods of probe labelling and detection were evaluated and found unsuitable for FISH because of tissue autofluorescence. Likewise, the use of avidin to detect biotin-labelled probe was obviated by the presence of endogenous biotin. An existing approach, using digoxigenin (DIG)-labelled probes and detection by anti-DIG antibody-labelled with alkaline phosphatase, was modified to use a fluorescent substrate, 2-hydroxy-3-naphthoic acid-2′-phenylanilide phosphate/4-chloro-2-methylbenzene diazonium hemi-zinc chloride salt (HNPP/Fast Red TR). This improved method allowed sensitive detection of IPNV target, without interference from autofluorescence or endogenous alkaline phosphatase. Furthermore, the reporter produces a discrete, non-fading signal, which is particularly suitable for analysis by confocal microscopy. [ABSTRACT FROM AUTHOR]

The ability of anti-MBP Atlantic salmon antibodies to bind to MBP was investigated using ELISA under a range of pHs (3–10) and osmolarities (100–1000 mOsml−1) to determine the optimum binding conditions required for Atlantic salmon antibodies. The pH optimum was between pH 7–8 with pH 7·4 giving the highest level of antibody binding. However, the decrease in optical density was in excess of 60% at pHs 6·0 and 9·0 and in excess of 99% at pHs of 3·0 and 10·0. The optimum osmolarity required for antibody binding was in the range 100–400 mOsml−1 with 330 mOsml−1 giving the highest level of antibody binding. Above 400 mOsml−1 there was a rapid decrease in antibody binding with the OD dropping by over 50% at all dilutions. This decrease was maintained at all osmolarities over 400 mOsml−1 [Copyright &y& Elsevier]

Goldsinny wrasse, Ctenolabrus rupestris (L.). were experimentally and naturally infected with Aeromonas salmonicida. Goldsinny wrasse were found to be susceptible to infection with A. salmonicida but significantly more resistant to infection than Atlantic salmon, Salmo salar L. The pathology of the acute infection is described. Following infection significantly higher levels of agglutinating antibody were detected in the sera of recovered wrasse than those observed in the Atlantic salmon controls. However, a large proportion of the recovered wrasse were culture positive for A. salmonicida and appeared to have entered a chronic stage of infection quite distinct from the carrier status that can develop in Atlantic salmon. This study indicates that, although goldsinny wrasse are susceptible to A. salmonicida, these fish are unlikely to become primarily infected, but contract furunculosis from infected Atlantic salmon. However, goldsinny wrasse may act as a reservoir for subsequent infections of cultivated Atlantic salmon because of the development of a chronic form of the disease. The potential for the vaccination of goldsinny wrasse against furunculosis is discussed. [ABSTRACT FROM AUTHOR]

Atlantic halibut (Hippoglossus hippoglossus L.) is a potentially important new species to cold-water aquaculture. Development of a viable industrial farming technique has been hampered by continued pathogen problems within the rearing cycle and there are several reports that indicated how susceptible juvenile halibut are to bacterial and viral diseases [1]. Interest has been expressed, within the industry, over the possibility of vaccinating suitably sized animals to protect against the more common aquaculture pathogens. Vibrio spp. are of particular concern due to their ubiquitous nature and the relatively frequent occurrence of these pathogens within marine aquaculture. We have previously investigated the susceptibility of Atlantic halibut to infection by Vibrio anguillarum and the efficacy of intraperitoneal injected delivery of a commercial vaccine in protecting against the disease [2]. Given the very high rate of protection offered by immunisation we wanted to investigate the effect of alternate routes of administration on the efficacy of the vaccine. [Copyright &y& Elsevier]

Serum lysozyme was measured in Atlantic halibut (Hippoglossus hippoglossus L.) kept in a range of different conditions that included ambient photoperiod and temperature and controlled photoperiod and temperature. There was no significant difference between animals held in ambient conditions of 6 degrees C and those held in controlled conditions of 12 degrees C. Similarly, there was no significant difference between animals maintained in a long day photoperiod and those in a short day photoperiod. However, there was a significant difference between summer and winter readings. Whilst this would indicate a link between season and the defence system, there appears to be no link with apparent entrainment to different photoperiods and serum lysozyme levels.

A European strain of viral haemorrhagic septicaemia virus (VHSV) isolated from wild-caught cod Gadus morhua (H16/7/95) was shown to cause clinical disease and mortality in excess of 80% in juvenile Atlantic cod when administered by the intra-peritoneal (i.p.) route. No virus was recovered from cod cohabiting with experimentally infected fish at a ratio of 1:1, and no VHSV-associated mortality was demonstrated following immersion infection. External signs of disease in cod were the presence of exophthalmia and ascites. Virus was identified as VHSV by enzyme-linked immunosorbent assay (ELISA) and was recovered from both brain and organ pools (kidney, liver and spleen) of 100% of i.p. infected cod mortalities. Virus was also detected using an indirect immunofluorescence test on tissue imprints of kidney, liver, spleen and brain taken from moribund fish. The fact that cod were not susceptible to VHSV following waterborne exposure raises important questions surrounding the propagation, maintenance and impact of a naturally occurring reservoir of virus in the marine environment.

Experimental bath challenge of juvenile and sub-adult Atlantic halibut with Vibrio anguillarum induced severe mortalities of 47 and 80%, respectively. However, animals vaccinated with a commercial V. anguillarum vaccine demonstrated excellent protection against the disease (100% RPS). This study also describes the gross pathology and histological changes associated with this infection. A loss of coordination, haemorrhage at the fin base and splenomegaly were frequent findings. Serum agglutinating activity demonstrated a rise following vaccination, the mean log2 titre rising from 3.8 to 8.4. This was associated with a significant rise in antibody-mediated complement killing ability of immune serum when compared to non-immune serum.