Renibacterium salmoninarum and Mycobacterium spp.: two bacterial pathogens present at low levels in wild brown trout (Salmo trutta fario) populations in Austrian rivers.

BACKGROUND: Renibacterium salmoninarum and Mycobacterium sp. are important bacterial pathogens of fish. R. salmoninarum is the causative agent of bacterial kidney disease, a Gram-positive bacterium mostly known for causing chronic infections in salmonid fish, while multiple species belonging to the Mycobacterium genus have been associated with mycobacteriosis in fish as well as in human. The objective of this study was to determine the prevalence of these two bacterial pathogens in populations of wild brown trout (Salmo trutta fario) in four rivers (Kamp, Wulka, Traun and Ybbs) in Austria. RESULTS: A total of 457 kidney samples were examined for both bacterial agents using nested and conventional PCR as well as bacterial cultivation on KDM-2, histological examination and immunohistochemistry. Molecular evidence showed an estimated prevalence level of 0.94% for R. salmoninarum in 2017 while the bacterium could not be detected in 2018 and histology showed signs consistent with a low-level chronic inflammation in the kidney of infected fish. Similarly, no fish were found positive for Mycobacterium in 2017 but in 2018, the prevalence was found to be 37.03% in the Kamp river (4.08% across all rivers). The sequencing data confirmed that these fish carried Mycobacterium sp. although the precise species of Mycobacterium could not be ascertained. CONCLUSIONS: This survey constitutes the first insight into the prevalence rate of R. salmoninarum and Mycobacterium sp. in wild brown trout (Salmo trutta fario) populations in Austria. Both of these pathogens were only detected in the summer months (June and July), which might suggest that the stress linked to increased water temperature could act as stressor factor and contribute to the outbreak of these diseases. The age of the fish might also play a role, especially in the case of Mycobacterium sp. as all the infected fish were in their first summer (June).

Susceptibility of common carp and sunfish to a strain of Francisella noatunensis subsp. orientalis in a challenge experiment.

Francisellosis, an emerging disease in many fish species, can cause high mortality in affected populations. Here we investigated the susceptibility of common carp Cyprinus carpio and sunfish Lepomis gibbosus to Francisella noatunensis subsp. orientalis (Fno), and possible transmission of the bacteria between the 2 fish species. In a challenge experiment, 3 groups of each species were injected intraperitoneally (IP) with 3 different doses of an Fno strain no. 9449 of the Norwegian Veterinary Institute, recovered from naturally infected ornamental Malawi cichlids. Infected carp were cohabitated with sunfish and vice versa. Control groups were injected with 0.9M phosphate-buffered saline and cohabitated accordingly. Fish were sampled at different time points. Mortality of challenged sunfish was observed during the first 96 h and reached 56.1%. In the control sunfish, 4 of 16 fish (25%) died within 48 h. In carp, no mortalities or clinical signs were observed during the experiment. General clinical and patho-anatomical disease signs of affected sunfish were observed. We detected granulomas in 2 cohabitated sunfish and 1 challenged carp, but could not re-isolate Fno from these fish. Fno was successfully cultured from 6 sunfish and 3 carp specimens until 35 d post injection. PCR of spleen and kidney with 16S rDNA Francisella-like bacterium primers 180f and 485r yielded amplicons in 68.3% of challenged sunfish and only 12.2% of challenged carp. We demonstrated that sunfish were susceptible to Fno infection while the carp were not. Horizontal transmission of the agent between the 2 fish species could not be demonstrated.

Effect of a phytogenic feed additive on the susceptibility of Onchorhynchus mykiss to Aeromonas salmonicida.

In recent years, feed additives have increasingly been adopted by the aquaculture industry. These supplements not only offer an alternative to antibiotics but have also been linked to enhanced growth performance. However, the literature is still limited and provides contradictory information on their effectiveness. This is mainly due to the wide variety of available products and their complex mechanisms of action. Phytogenic feed additives have been shown to have antimicrobial effects and can improve growth performance. In the present study, we investigated the susceptibility of several fish pathogenic bacteria to a phytogenic essential oil product in vitro. In addition, we determined the protective effect of a commercial phytogenic feed additive containing oregano, anis and citrus oils on the resistance of rainbow trout Oncorhynchus mykiss to infection by Aeromonas salmonicida. The bacterium was administered through 3 different routes: intra-peritoneal injection, immersion in a bacterial solution and cohabitation with infected fish. Mortality rates were significantly lower in infected rainbow trout that had received the feed additive: the overall mortality rate across all routes of infection was 18% in fish fed a diet containing the additive compared to 37% in fish that received unsupplemented feed. The route of infection also significantly impacted mortality, with average mortality rates of 60, 17.5 and 5% for intra-peritoneal injection, immersion and cohabitation, respectively. In general, fish were better protected against infection by immersion than infection by injection.