Growth indices were examined in 24 identically managed tanks, each containing 120 diploid juvenile rainbow trout (initial mean body weight, 9.3 g), during a 12-week study to examine tank effects associated with tank location in a multi-user research facility. Growth indices included mean body weight, feed intake, feed conversion index, and specific growth rate. The null hypothesis that tank effect had no effect on growth over the 12-week period was rejected (P = 0.038), and mean weight in individual tanks differed by as much as 18.7%. During the study, it was determined that the proximity of tanks to common-use walkways in the facility could affect growth indices. This was indicated by significant differences in the mean fish weights among blocks of tanks served by different header tanks after 4 (P = 0.001) and 8 (P = 0.024) weeks. The block containing tanks of fish with the highest mean weight was nearest to the 2 common-use walkways in the facility. Fish in this block of tanks, compared with those in other blocks, had significantly greater feed intake but no significant differences in conversion efficiency. Compensatory growth, a well known growth attribute in fishes, diminished the difference in mean weight between these blocks of tanks by the end of the study. Comparison of paired tanks within header tank blocks indicated that fish in those located nearest to walkways had higher feeding rates over the 12-week period (P = 0.048), but less efficient teed conversion (P = 0.040) than did fish in matched tanks located farthest from walkways. However, there were no differences in mean weight of fish. Results of this trial document the risks involved in identifying fish in a tank as the experimental unit when treatments are administered to the tank of fish, the latter being the true experimental unit.

The Shewanella putrefaciens group are ubiquitous microorganisms recently isolated from different freshwater fish species and causing serious health disorders. The purpose of the study was to characterise isolates of the S. putrefaciens group with special emphasis on elucidating serological diversity and determining putative virulence factors. Isolates collected from freshwater fish (n = 44) and reference strains were used. The identification of bacteria was carried out using biochemical kits and 16S rRNA sequencing. Polyclonal antibodies were prepared against the S. putrefaciens group. The bacterium’s susceptibility to antimicrobial agents, its enzymatic properties, and its adhesion ability to fish cell lines were also tested. Finally, selected isolates were used in challenge experiments in common carp and rainbow trout. Excluding six isolates undeterminable for species, the bacteria were classified to three species: S. putrefaciens, S. xiamenensis, and S. oneidensis, and showed some phenotypic diversity. Fourteen serological variants of the S. putrefaciens group were determined with the newly developed serotyping scheme. Serodiversity may play an important role in the virulence of particular isolates. Further, S. putrefaciens group members adhere to epithelial cells and produce enzymes which may contribute to their virulence. Challenge tests confirmed the pathogenicity of the S. putrefaciens group for fish.

Introduction: The goal of this study was to assess the influence of vaccination against enteric redmouth disease on oxidative stress biomarkers and antioxidant defence in the muscle tissue of rainbow trout (Oncorhynchus mykiss Walbaum) vaccinated against Yersinia ruckeri in the first and second month after immunisation. Material and Methods: Healthy fish were vaccinated orally with inactivated whole cells of a virulent strain of Y. ruckeri. One and two months after immunisation the muscle samples were collected. Results: No significant difference was noted in lipid peroxidation level in either the first or second month after vaccination, while aldehydic and ketonic derivatives of oxidatively modified proteins (OMB) in the vaccinated group were significantly lower in the second month compared to those in the first month after vaccination (P < 0.05). The content of ketonic derivatives of OMB in muscles in the first month after immunisation was higher compared to untreated control. All these culminated in a depletion of glutathione peroxidase (GPx) activity and low level of total antioxidant capacity (TAC). Conclusion: Correlations between catalase activity and lipid peroxidation and TAC confirmed the pivotal role of catalase in antioxidant defence during immunisation. From a broader perspective, it is suggested that immunisation of fish with Yersinia vaccine is associated with induced free radical formation and oxidative stress. Free radicals would therefore be at least partially responsible for the induction of both humoral and cellular elements of the immunity and increased protective immunity against Y. ruckeri infection.

Based on analysis of available genome sequences, five gene lineages of MHC class I molecules (MHC I-U, -Z, -S, -L and -P) and one gene lineage of MHC class II molecules (MHC II-D) have been identified in Osteichthyes. In the latter lineage, three MHC II molecule sublineages have been identified (MHC II-A, -B and -E). As regards MHC class I molecules in Osteichthyes, it is important to take note of the fact that the lineages U and Z in MHC I genes have been identified in almost all fish species examined so far. Phylogenetic studies into MHC II molecule genes of sublineages A and B suggest that they may be descended from the genes of the sublineage named A/B that have been identified in spotted gar (Lepisosteus oculatus). The sublineage E genes of MHC II molecules, which represent the group of non-polymorphic genes with poor expression in the tissues connected with the immune system, are present in primitive fish, i.e. in paddlefish, sturgeons and spotted gar (Lepisosteus oculatus), as well as in cyprinids (Cyprinidae), Atlantic salmon (Salmo salar), and rainbow trout (Oncorhynchus mykiss). Full elucidation of the details relating to the organisation and functioning of the particular components of the major histocompatibility complex in Osteichthyes can advance the understanding of the evolution of the MHC molecule genes and the immune mechanism.

This study describes the macroscopic and microscopic lesions in farmed rainbow trout (Oncorhynchus mykiss) naturally infected with genetically identified Aeromonas salmonicida, A. hydrophila, and A. veronii species. The genus Aeromonas includes bacteria that naturally inhabit both waterways and organisms. At least 27 Aeromonas species have been identified to date, some of which can cause significant economic losses in aquaculture. As up to 68.8% of Aeromonas isolates may be misidentified in routine biochemical and phenotypic tests, however, reported cases of Aeromonas infection in fish may be wrongly identified. Our findings confirmed that the 3 Aeromonas species studied are associated with septicemia and dermal lesions in rainbow trout.

Microcystin and related toxic peptides produced by cyanobacteria (blue-green algae) are potent and selective inhibitors of protein phosphatases 1 and 2A. We adapted existing enzymatic techniques to analyze the liver of rainbow trout after oral administration of hepatotoxic cyanobacteria. Liver tissue was removed 3 and 12 hours after treatment, and phosphatase activity was determined in liver extracts, using a specific phosphoprotein substrate. In all samples from fish exposed to toxic cyanobacteria, phosphatase activity was suppressed, whereas the control enzyme, lactate dehydrogenase, present in the same liver extract, was not affected by cyanobacteria. Thus, experimental poisoning by hepatotoxic cyanobacteria resulted in an abnormally low ratio of phosphatase to lactate dehydrogenase activity in the liver extracts. These results indicate that specific inhibition of phosphatases 1 and 2A may provide a useful diagnostic tool to determine the early effects of cyanobacteria toxic peptides directly in liver samples from poisoned animals. Although this test was developed with rainbow trout, it should be possible to extend the analysis of liver phosphatase activity to other species, including sheep and cattle, which are frequently affected by hepatotoxic cyanobacteria.

Inclusion of high-ionic strength buffers helped us to develop a sandwich ELISA to detect hemorrhagic septicemia virus (HSV) in cell culture and infected trout tissue extracts. For maximal sensitivity of 0.1 to 0.2 ng/well/100 microliter or about 10 to 50 TCID50/well/100 microliter, trout extracts were diluted 1:1 and assayed for the earliest synthesized nucleoprotein N. Simultaneous binding of the N protein from HSV in the sample to the wells coated with monoclonal antibody (2D5 against the N protein) and to the peroxidase-labeled monoclonal antibody (2C9 against the N protein) proceeded during a 2-hour incubation at 20 to 22 C (room temperature). The response was linear between 6 to 60 ng/well of purified virus. Monoclonal antibodies used were noncompetitive with each other and reacted with F1, F2, 23.75, and 5 Spanish isolates of HSV, but not with infectious hematopoietic necrosis or infectious pancreatic necrosis viruses. Tissue specimens with low content of HSV virus may now be assayed directly without use of cell culture, rapidly, and with high precision, during the acute phase of the disease in salmonid fishes.

Growth indices were examined in 24 identically managed tanks, each containing 120 diploid juvenile rainbow trout (initial mean body weight, 9.3 g), during a 12-week study to examine tank effects associated with tank location in a multi-user research facility. Growth indices included mean body weight, feed intake, feed conversion index, and specific growth rate. The null hypothesis that tank effect had no effect on growth over the 12-week period was rejected (P = 0.038), and mean weight in individual tanks differed by as much as 18.7%. During the study, it was determined that the proximity of tanks to common-use walkways in the facility could affect growth indices. This was indicated by significant differences in the mean fish weights among blocks of tanks served by different header tanks after 4 (P = 0.001) and 8 (P = 0.024) weeks. The block containing tanks of fish with the highest mean weight was nearest to the 2 common-use walkways in the facility. Fish in this block of tanks, compared with those in other blocks, had significantly greater feed intake but no significant differences in conversion efficiency. Compensatory growth, a well known growth attribute in fishes, diminished the difference in mean weight between these blocks of tanks by the end of the study. Comparison of paired tanks within header tank blocks indicated that fish in those located nearest to walkways had higher feeding rates over the 12-week period (P = 0.048), but less efficient teed conversion (P = 0.040) than did fish in matched tanks located farthest from walkways. However, there were no differences in mean weight of fish. Results of this trial document the risks involved in identifying fish in a tank as the experimental unit when treatments are administered to the tank of fish, the latter being the true experimental unit.