Susceptibility of Caligus rogercresseyi collected from the native fish species Eleginops maclovinus (Cuvier) to antiparasitics applied by immersion

A major challenge for Chilean salmon farming is infestation by the ectoparasite Caligus rogercresseyi. In addition, there is evidence that a loss of chemotherapeutic treatment efficacy against important fish pathogens is occurring in salmon farming, including antiparasitic efficacy. Currently, there are known techniques that allow the determination of the susceptibility profile of parasites to antiparasitic treatment. However, there is scarce information about both threshold values and categorization of antiparasitic susceptibility for C. rogercresseyi. Bioassay technique allowed the determination of both mean values and the natural variation of EC50%, which were contrasted with available susceptibility thresholds. Results allowed to determine that parasites from the native fish host, Eleginops maclovinus, are susceptible to azamethiphos, deltamethrin and cypermethrin treatments, showing a high susceptibility profile to antiparasitics.     

Presence and genetic variability of Piscine orthoreovirus genotype 1 (PRV‐1) in wild salmonids in Northern Europe and North Atlantic Ocean

Piscine orthoreovirus genotype 1 (PRV‐1) is widespread in farmed Atlantic salmon (Salmo salar L.) populations in northern Europe, Canada and Chile. PRV‐1 occurs in wild fish in Norway and Canada; however, little information of its geographical distribution in wild populations is currently available, and the effect of PRV‐1 infection in wild populations is currently unknown. In this study, we present the findings of a survey conducted on 1,130 wild salmonids sampled in Denmark, Sweden, Ireland, Faroe Islands, France, Belgium and Greenland between 2008 and 2017. PRV‐1 is reported for the first time in wild salmonids in Denmark, Sweden, Faroe Island and Ireland. The annual PRV‐1 prevalence ranged from 0% in France, Belgium and Greenland to 43% in Faroe Islands. In total, 66 samples tested positive for PRV‐1, including Atlantic salmon broodfish returning to spawn and Atlantic salmon collected at the feeding ground north of Faroe Islands. The phylogenetic analysis of S1 sequences of the PRV‐1 isolates obtained in this survey did not show systematic geographical distribution. This study sheds light on the spread and genetic diversity of the virus identified in populations of free‐living fish and provides rationale for screening wild broodfish used in restocking programmes.     

Transmission of Mycobacterium chelonae and Mycobacterium marinum in laboratory zebrafish through live feeds

The zebrafish (Danio rerio) is a popular vertebrate model organism used in a wide range of research fields. Mycobacteriosis, caused by Mycobacterium species, is particularly concerning because it is a common disease associated with chronic infections in these fish. Infections are also a source of uncontrolled experimental variance that may influence research results. Live feeds for zebrafish are common and include paramecia (Paramecium caudatum), brine shrimp (Artemia franciscana) and rotifers (Branchionus spp.). Although nutritionally beneficial, live feeds may pose a biosecurity risk. In this study, we investigate transmission of Mycobacterium chelonae and Mycobacterium marinum through these three live feeds. We show that all three live feeds ingest both M. marinum and M. chelonae and can transmit mycobacterial infections to zebrafish. This observation emphasizes the need for live feeds to be included in the consideration of potential biosecurity risks. This study is of importance to other beyond the zebrafish community, including those of additional aquatic models and those using live feeds for other types of aquaculture.     

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Vibrio ordalii is an extracellular, Gram‐negative bacterium that produces vibriosis in salmonids. While pathogenesis is not fully understood, this bacterium has numerous likely genes for adhesion, colonization, invasion factors and, as recently suggested, intracellular behaviour. Therefore, this study aimed to clarify possible intracellular behaviour for V. ordalii Vo‐LM‐18 and ATCC 33509^T in the fish‐cell lines SHK‐1 and CHSE‐214. Confocal microscopy revealed Vo‐LM‐18 and ATCC 33509^T inside cytoplasm in both fish‐cell lines at 4 hr post‐inoculation (hpi). At 8 and 16 hpi, the proportion of fish cells invaded by both strains increased. Moreover, intracellular V. ordalii were observed after 8 hpi inside mouse embryonic fibroblasts (MEF), demonstrating that entry was not due to a cellular phagocytosis process. Flow cytometry confirmed immunocytochemistry results, with both V. ordalii evidencing statistically significant differences in the number of infected cells between 8 and 16 hpi. Interestingly, V. ordalii infection did not significantly damage fish cells, as determined by LDH liberation. Viable counts at 8 hpi detected, on average for both lines, 176 ± 47 CFU/ml of culturable intracellular Vo‐LM‐18 and ATCC 33509^T cells. These in vitro findings support the facultative intracellular behaviour of V. ordalii and may be of importance for understanding pathogenicity and survival in aquatic environments.     

To pool or not to pool? Guidelines for pooling samples for use in surveillance testing of infectious diseases in aquatic animals

Samples from multiple animals may be pooled and tested to reduce costs of surveillance for infectious agents in aquatic animal populations. The primary advantage of pooling is increased population‐level coverage when prevalence is low (<10%) and the number of tests is fixed, because of increased likelihood of including target analyte from at least one infected animal in a tested pool. Important questions and a priori design considerations need to be addressed. Unfortunately, pooling recommendations in disease‐specific chapters of the 2018 OIE Aquatic Manual are incomplete and, except for amphibian chytrid fungus, are not supported by peer‐reviewed research. A systematic review identified only 12 peer‐reviewed aquatic diagnostic accuracy and surveillance studies using pooled samples. No clear patterns for pooling methods and characteristics were evident across reviewed studies, although most authors agreed there is a negative effect on detection. Therefore, our purpose was to review pooling procedures used in published aquatic infectious disease research, present evidence‐based guidelines, and provide simulated data examples for white spot syndrome virus in shrimp. A decision tree of pooling guidelines was developed for use by peer‐reviewed journals and research institutions for the design, statistical analysis and reporting of comparative accuracy studies of individual and pooled tests for surveillance purposes.     

Growth and survival of juvenile red drum,Sciaenops ocellatus,acclimated to freshwater at three different stocking densities in a partial recirculation system

Preliminary technical baseline information for the inland culture of juvenile red drum, Sciaenops ocellatus, in a freshwater fish hatchery is presented. Three initial stocking densities (d₁ = 83, d₂ = 167, and d₃ = 250 fish/m³) were tested in a semirecirculation system for 575 days using a commercially available feed (48.1% protein, 25% lipids, and 0.13% fiber). Initial and final mean fish weight were 7.11 ± 0.02 g and 287.6 ± 27 g. As carrying capacity appeared to have been reached, only data from Days 1–333 were analyzed. One‐way ANOVA analyses indicated that the survival rate of the smaller density (d₁ =40 ± 3.7%) was significantly different from the other two densities (d₃ = 21 ± 1.1% and d₂ = 23 ± 1.0%), which did not differ between them. Absolute growth rate (0.87 ± 0.02 g/day) was not significantly different between densities (F = 0.23. p = .801). Neither was weight significantly different between treatments. There were significant differences in the feed conversion rate (FCR) between densities (F = 8.54; p = .02). FCR for d₁ was significantly lower than for the two other densities, which did not differ from each other. A von Bertalanffy growth model was adjusted (R² = 0.95), and weight–length relationship presented negative allometric values (b = 2.85, R² = 0.98).     

Comparative effect of advanced soy products or corn protein concentrate with porcine meal on growth,body composition,and distal intestine histology of Florida pompano,Trachinotus carolinus

The present study was designed to investigate the effects of diets containing advanced soy products (enzyme‐treated soy and fermented soy) or corn protein concentrate (CPC) in combination with porcine meal (PM) to completely replace poultry byproduct meal (PBM) on growth performance, body composition, and distal intestine histology of Florida pompano, Trachinotus carolinus. Four experimental diets were formulated to be isonitrogenous and isolipidic, to contain 400 g/kg crude protein and 80 g/kg lipid. A reference diet (PBM diet [PBMD]) contained 150 g/kg PBM and 495 g/kg soybean meal (SBM), and three test diets were formulated replacing PBM with 15 g/kg of CPC (CPC diet [CPCD]) or replacing all SBM and PBM with 535 g/kg fermented soy (fermented soybean meal diet [FSBMD]) or 451.3 g/kg enzyme‐treated soy (enzyme‐treated soybean meal diet [ESBMD]). All three test diets were supplemented with 38 g/kg of PM. Diets were fed based on a percentage of bodyweight adjusted after sampling the fish every 2 weeks to triplicate groups of Florida pompano juveniles (mean weight 8.06 ± 0.22 g). After 8 weeks of feeding, fish fed CPCD and ESBMD performed equally well in terms of final body weight, thermal growth coefficient, and percentage weight gain in comparison to fish fed PBMD. In all cases, feeding FSBMD resulted in poor feed conversion and lower feed intake compared to other treatments. Protein retention efficiency, whole‐body proximate composition, phosphorus, sulfur, potassium, magnesium, calcium, sodium, and zinc contents were not significantly influenced by the dietary treatments. The results obtained in the present histological study showed no significant differences in the thickness of serous layer, muscular layer, and submucosal layer of the intestine among treatments. Fish fed CPCD showed a significant widening of the lamina propria with an increase of cellular infiltration and higher presence of goblet cells compared to other dietary treatment. Based on these results, 451 g/kg ESBM or combination of 150 g/kg of CPC and 495 g/kg SBM supplemented with 38 g/kg PM can be utilized to develop a practical diet for juvenile Florida pompano without impacting growth, nutritive parameters, and several distal intestine health parameters.     

Replacement of fish oil in plant‐based diets for Pacific white shrimp,Litopenaeus vannamei,by stearine fish oil and palm oil

Stearine fish oil (SFO) and palm oil (PO) have emerged as promising alternatives for the replacement of fish oil (FO) in aquafeeds. This study evaluated the replacement of FO with alternative oils in practical diets for Litopenaeus vannamei. In a clear brackish water study (14.1 g/L) utilizing shrimp (0.29 ± 0.02 g, initial weight), FO was replaced by SFO at inclusion ratios of 100:0, 75:25, 50:50, 25:75, and 0:100 (FO:SFO) and PO as 90% of FO. After 55 days, no significant differences (p < 0.05) in final weight, growth, or survival of shrimp were observed. A second trial (8 weeks) in low‐salinity water (2.1 g/L) with shrimp (0.92 ± 0.02 g, initial weight) evaluated diets with 100% FO, 100% SFO, 90% PO, 90% soybean oil (SO), or 90% flaxseed oil (FXO) as a replacement for FO and four commercially produced diets with 2% of FO, SO, PO, or FXO. One treatment received half rations of the commercial FO diet, and one treatment was based entirely on natural productivity. Results show that the fatty acid profiles of the tail muscle conformed to the lipids of the feed, and highly unsaturated fatty acids (HUFAs) were preserved. Following 8 weeks of culture, there were no significant differences in production performance.