Pharmacokinetics, optimal dosages and withdrawal time of florfenicol in Asian seabass (Lates calcarifer) after oral administration via medicated feed.

Asian seabass (Lates calcarifer) is an economically important fish in Asian and Australian markets, but few pharmacokinetic (PK) data of antimicrobial drugs in this species is available. The present study investigated the PK behaviour of florfenicol (FF) through medicated feed in Asian seabass cultured at 25°C. The serum and muscle/skin concentrations of FF and its metabolite florfenicol amine (FFA) were determined by the HPLC-FLD method and analysed by one-compartmental model. The optimal dosages were determined by pharmacokinetic-pharmacodynamic (PK-PD) approach and the linear regression analysis was used to determine the withdrawal time (WDT). The PK study following a single oral administration of 15 mg/kg FF via medicated feed revealed that the absorption half-life (t1/2Ka ), elimination half-life (t1/2K ), peak concentration (Cmax ), area under the concentration-time curve (AUC), volume of distribution (Vd/F) and clearance (CL/F) were 1.47 h, 8.07 h, 8.61 µg/ml, 146.41 h·µg/ml, 1.19 L/kg and 0.102 L/kg/h, respectively. The muscle/skin concentration-time profile was similar to that of the serum, suggesting well distribution but only a small fraction of FF was metabolized to FFA. The optimal dosage for a minimum inhibitory concentration of 2 µg/ml was calculated as 13.38 mg/kg/day. The appropriate WDT after multiple oral medications with 15 mg/kg FF once daily for 7 days was determined as 8 days. Information obtained from the current study can potentially be applied for the treatment of bacterial diseases in farming Asian seabass.

Square root identities for harvested Beverton-Holt models.

We introduce the term net-proliferation rate for a class of harvested single species models, where harvest is assumed to reduce the survival probability of individuals. Following the classical maximum sustainable yield calculations, we establish relations between the proliferation and net-proliferation that are economically and sustainably favored. The resulting square root identities are analytically derived for species following the Beverton-Holt recurrence considering three levels of complexity. To discuss the generalization of the results, we compare the square root result to the optimal survival rate of the Pella-Tomlinson model. Furthermore, to test the practical relevance of the square root identities, we fit a stochastic Pella-Tomlinson model to observed Barramundi fishery data from the Southern Gulf of Carpentaria, Australia. The results show that for the estimated model parameters, the equilibrium biomass levels resulting from the MSY harvest and the square root harvest are similar, supporting the claim that the square root harvest can serve as a rule-of-thumb. This application, with its inherited model uncertainty, sparks a risk sensitivity analysis regarding the probability of populations falling below an unsustainable threshold. Characterization of such sensitivity helps in the understanding of both dangers of overfishing and potential remedies.

Environmental drivers of recruitment in a tropical fishery: Monsoonal effects and vulnerability to water abstraction.

Fisheries and natural water resources across the world are under increasing pressure from human activity, including fishing and irrigated agriculture. There is an urgent need for information on the climatic/hydrologic drivers of fishery productivity that can be readily applied to management. We use a generalized linear mixed model framework of catch curve regression to resolve the key climatic/hydrological drivers of recruitment in Barramundi Lates calcarifer using biochronological (otolith aging) data collected from four river-estuary systems in the Northern Territory, Australia. These models were then used to generate estimates of the year class strength (YCS) outcomes of different water abstraction scenarios (ranging from 10% to 40% abstraction per season/annum) for two of the rivers in low, moderate, and high discharge years. Barramundi YCS displayed strong interannual variation and was positively correlated with regional monsoon activity in all four rivers. River-specific analyses identified strong relationships between YCS and several river-specific hydrology variables, including wet and dry season discharge and flow duration. Water abstraction scenario models based on YCS-hydrology relationships predicted reductions of >30% in YCS in several cases, suggesting that increased water resource development in the future may pose risks for Barramundi recruitment and fishery productivity. Our study demonstrates the importance of the tropical monsoon as a driver of Barramundi recruitment and the potential for detrimental impacts of increased water abstraction on fishery productivity. The biochronological and statistical approaches we used have the potential to be broadly applied to inform policy and management of water resource and fisheries.

Influence of life history variation and habitat on mercury bioaccumulation in a high-order predatory fish in tropical Australia.

Mercury distribution and bioaccumulation in aquatic ecosystems of tropical Australia is poorly characterised. Barramundi (Lates calcarifer), a widespread high-order predator in both fresh and coastal marine waters of the region, fulfils requirements for a bio-indicator of mercury contamination. In a study of the Mary River system of the Northern Territory, total mercury in the muscle tissue of 300 specimens gathered over four years (2013-2017, across both wet and dry seasons) was determined by direct combustion-atomic absorption spectrometry. Source of nutrition and trophic position of barramundi in the food web was also estimated via carbon and nitrogen isotopes (δ13C and δ15N), respectively, in tissue by stable isotope mass spectrometry, and determination of strontium isotopes (87Sr/86Sr) in otoliths by laser ablation-ICPMS differentiated between freshwater and saltwater residence. Results showed that fish moving into freshwater floodplain wetlands concentrated mercury in muscle tissue at approximately twice the level of those that remained in saline habitats. Resolving life histories through otolith analyses demonstrated diversity in mercury bioaccumulation for individual fish of the same migratory contingent on the floodplains. Although trophic level (δ15N), capture location, source of nutrition (δ13C), and age or size partly predicted mercury concentrations in barramundi, our results suggest that individual variability in diets, migration patterns and potentially metabolism are also influential. Using a migratory fish as a bio-indicator, and tracking its life history and use of resources, proved valuable as a tool to discern hot spots in a coastal waterway for a contaminant, such as mercury.

Thermal performance curves for aerobic scope in a tropical fish (Lates calcarifer): flexible in amplitude but not breadth.

Aerobic metabolic scope is a popular metric to estimate the capacity for temperature-dependent performance in aquatic animals. Despite this popularity, little is known of the role of temperature acclimation and variability in shaping the breadth and amplitude of the thermal performance curve for aerobic scope. If daily thermal experience can modify the characteristics of the thermal performance curve, interpretations of aerobic scope data from the literature may be misguided. Here, tropical barramundi (Lates calcarifer) were acclimated for ∼4 months to cold (23°C), optimal (29°C) or warm (35°C) conditions, or to a daily temperature cycle between 23 and 35°C (with a mean of 29°C). Measurements of aerobic scope were conducted every 3-4 weeks at three temperatures (23, 29 and 35°C), and growth rates were monitored. Acclimation to constant temperatures caused some changes in aerobic scope at the three measurement temperatures via adjustments in standard and maximum metabolic rates, and growth rates were lower in the 23°C-acclimated group than in all other groups. The metabolic parameters and growth rates of the thermally variable group remained similar to those of the 29°C-acclimated group. Thus, acclimation to a variable temperature regime did not broaden the thermal performance curve for aerobic scope. We propose that thermal performance curves for aerobic scope are more plastic in amplitude than in breadth, and that the metabolic phenotype of at least some fish may be more dependent on the mean daily temperature than on the daily temperature range.

PCR, in-situ hybridization, and phylogenetic analysis suggest that 'big belly' disease in barramundi, Lates calcarifer (Bloch), is caused by a novel Vibrio species.

'Big belly' disease is a chronic, granulomatous bacterial enteritis and peritonitis, first reported in 3- to 4-week-old Asian seabass or barramundi, Lates calcarifer Bloch fry. Affected fry are emaciated and have a swollen abdomen, and the condition is referred to as 'skinny pot-belly' or 'big belly' disease. In this study, histopathological examinations of diseased fish from a batch of 2-month-old, 6- to 8-cm L. calcarifer fingerlings, kept in seawater recirculating aquaculture systems, showed pathology resembling 'big belly' disease. Ethanol-fixed tissues were tested positive using specific PCR primers based on 16S rRNA genes. In situ hybridization using dioxygenin-labelled positive PCR products on formalin-fixed paraffin-embedded tissues showed positive reactions with intralesional, clusters of the large, 'big belly' coccobacilli. A phylogenetic tree constructed based on analyses of these 16S rRNA gene PCR products from five positive fish suggests that the 'big belly' bacterium is most likely a novel Vibrio species.

Tracking the resource pulse: Movement responses of fish to dynamic floodplain habitat in a tropical river.

Natural river floodplains are among the Earth's most biologically diverse and productive ecosystems but face a range of critical threats due to human disturbance. Understanding the ecological processes that support biodiversity and productivity in floodplain rivers is essential for their future protection and rehabilitation. Fish assemblage structure on tropical river floodplains is widely considered to be driven by dispersal limitation during the wet season and by environmental filtering and interspecific interactions during the dry season. However, the individual-level movement behaviours (e.g. site attachment, nomadism, homing) that regulate dispersal of fish on floodplains are poorly understood. We combined radiotelemetry and remote sensing to examine the movement behaviour of two large-bodied fishes (barramundi Lates calcarifer, forktail catfish Neoarius leptaspis) over the flood cycle in a tropical river-floodplain system in northern Australia to: (a) quantify movement responses in relation to dynamic habitat resources at a landscape scale; and (b) determine the extent of spatial 'reshuffling' of individual fish following the wet season. Both species altered their behaviour rapidly in response to changes in the availability and distribution of aquatic habitat, with most individuals undertaking extensive movements (up to ~27 km from the tagging location) on the inundated floodplain during the wet season. Although there was considerable individual variation in movement patterns, overall barramundi distributions closely tracked the extent of floodplain primary productivity, whereas catfish distributions were most closely associated with the extent of flooded area. Most individuals of both species exhibited homing back to previously occupied dry season refugia during the wet-to-dry transition, even though other potential refugia were available in closer proximity to wet season activity areas. We postulate that homing behaviour modulates temporal variation in fish assemblage composition and abundance and limits the transfer of aquatic-derived energy and nutrients into terrestrial food webs by reducing fish mortality on drying floodplains. Our study demonstrates the importance of quantifying individual-level behaviour across the three stages of dispersal (emigration, inter-patch movement, immigration) for our understanding of how animal movement influences energetic subsidies and other large-scale ecosystem processes.

The ameliorative effects of various fish protein hydrolysates in poultry by-product meal based diets on muscle quality, serum biochemistry and immunity in juvenile barramundi, Lates calcarifer.

In an effort to reduce the use of fishmeal (FM), the effect of using protein from poultry by product meal (PBM) along with the supplementation of three different fish protein hydrolysate (FPH) including yellowtail kingfish, carp and tuna hydrolysate (designated as KH, CH and TH, respectively) were evaluated in juvenile barramundi for growth performance, fillet quality, mucosal immunity, serum biochemistry, immune response and infection against Vibrio harveyi. Fish were fed a FM based control diet + three isonitrogenous and isolipidic diets containing 90% of PBM protein supplemented with different types of hydrolysates: 90% PBM +10% KH (90PBM + KH), 90% PBM + 10% CH (90PBM + CH) and 90% PBM + 10% TH (90PBM + TH). Growth performance and indices were unaffected by the hydrolysate supplemented diets when compared to the control. FPH supplemented PBM diets resulted in improved muscle quality by improving poly unsaturated fatty acids (PUFA), ∑n-3, ∑n-6 and ∑n-9, and health related lipid indexes were not affected. The internal architecture of spleen and kidney were not altered by test diets whilst FPH supplemented PBM modulated acidic mucins in intestine and skin of fish. Improved infection rate in response to two weeks post infection with V. harveyi in the FPH supplemented diets was further associated with an increased serum immune response and a concomitant regulation of proinflammatory and inflammatory cytokines in the head kidney. Serum biochemistry including alanine transaminase (ALT), glutamate dehydrogenase (GLDH) and total bilirubin (TB) showed a decreasing trend both in pre-challenge and post-challenge barramundi fed FPH supplemented diets whereas cholesterol level decreased significantly in post-challenge groups fed 90PBM + KH and 90PBM + TH than pre-challenge barramundi. This study signifies that supplementation of 10% with different three FPH, hydrolysed by an alcalase® enzyme in PBM-based diets for barramundi could be good strategies to overcome the negative consequences triggered by animal by-product ingredients.

Simultaneous detection of scale drop disease virus and Flavobacterium columnare from diseased freshwater-reared barramundi Lates calcarifer.

Freshwater farming of barramundi Lates calcarifer in Thailand is a growing sector in aquaculture, but mortalities due to infectious diseases are still a major threat to this industry. In 2018, an episode of severe mortality in juvenile barramundi was noted in a freshwater earth pond site. Fish presented with severe gill necrosis, as well as severe cutaneous hemorrhages, scale loss, and discoloration at the base of dorsal fin (saddleback lesions). Histopathology revealed extensive necrosis of skeletal muscle and gill filaments, as well as basophilic inclusion bodies and megalocytosis in muscle, gill, liver, and kidney. Scale drop disease virus (SDDV) infection was subsequently confirmed by virus-specific semi-nested PCR. Further, DNA sequences of the viral major capsid protein (MCP) and ATPase genes had a respective homology of 99.85 and 99.92% with sequences of SDDV infecting barramundi in saltwater culture. Gill necrosis and saddleback lesions are not typical lesions associated with scale drop syndrome. Their presence was explained by Flavobacterium columnare isolation from the gill, followed by positive F. columnare-specific PCR. To our knowledge, this is the first report of SDDV-associated mortality in freshwater-farmed barramundi. Furthermore, this mortality presented as a concurrent infection with SDDV and F. columnare, with typical lesions of both infections.

Henneguya (Cnidaria: Myxosporea: Myxobolidae) infections of cultured barramundi, Lates calcarifer (Perciformes: Latidae) in an estuarine wetlands system of Malaysia: description of Henneguya setiuensis n. sp., Henneguya voronini n. sp. and Henneguya calcarifer n. sp.

Examination of 35 barramundi (Lates calcarifer) from aquaculture cages in Setiu Wetland, Malaysia, revealed a single fish infected with three Henneguya spp. (Cnidaria: Myxosporea). Characterization of the infections using tissue tropism, myxospore morphology and morphometry and 18S rDNA sequencing supported description of three new species: Henneguya setiuensis n. sp., Henneguya voronini n. sp. and H. calcarifer n. sp. Myxospores of all three species had typical Henneguya morphology, with two polar capsules in the plane of the suture, an oval spore body, smooth valve cell surfaces, and two caudal appendages. Spores were morphometrically similar, and many dimensions overlapped, but H. voronini n. sp. had shorter caudal appendages compared with H. calcarifer n. sp. and H. setiuensis n. sp. Gross tissue tropism distinguished the muscle parasite H. calcarifer n. sp. from gill parasites H. setiuensis n. sp. and H. voronini n. sp.; and these latter two species were further separable by fine-scale location of developing plasmodia, which were intra-lamellar for H. setiuensis n. sp. and basal to the filaments for H. voronini n. sp. small subunit ribosomal DNA sequences distinguished all three species: the two gill species H. setiuensis n. sp. and H voronini n. sp. were only 88% similar (over 1708 bp), whereas the muscle species H. calcarifer n. sp. was most similar to H. voronini n. sp. (98% over 1696 bp). None of the three novel species was more than 90% similar to any known myxosporean sequence in GenBank. Low infection prevalence of these myxosporeans and lack of obvious tissue pathology from developing plasmodia suggested none of these parasites are currently a problem for barramundi culture in Setiu Wetland; however additional surveys of fish, particularly at different times of the year, would be informative for better risk assessment.

Adequate supply of dietary taurine stimulates expression of molecular markers of growth and protein turnover in juvenile barramundi (Lates calcarifer).

A trial was conducted to investigate the effect of dietary taurine (Tau) supply on the plasma amino acid composition and hepatic expression of several genes in juvenile barramundi (Lates calcarifer) after feeding. Triplicate tanks of fish (average weight, 89.3 g) were fed diets containing either a deficient (1 g kg-1), adequate (8 g kg-1) or excessive (19 g kg-1) level of dietary Tau. Liver tissues collected before feeding, and at 2- and 4-h post-feeding, were analysed for expression of genes involved in pathways of sulphur amino acid turnover, Tau biosynthesis and transport, target of rapamycin (TOR) signalling, the somatotropic axis and protein turnover. The treatment had no significant effect on the profiles of any amino acid in plasma collected over time after feeding, other than Tau and glycine. The expression profile of cystine and Tau synthetic genes suggested an effect of Tau excess on the metabolism of cystine. Markers of two pathways of Tau biosynthesis appear to be active in this species, providing proof that this species possesses the ability to synthesise Tau from SAA precursors. A marker for the regulation of Tau transport and homeostasis was shown to be directly regulated by Tau availability, whilst a link between adequate supply of Tau and TOR pathway-mediated growth stimulation was also apparent. An observed depression in expression of genes of the somatotropic axis, coupled with upregulation of the proteolytic and TOR-suppressing genes, in response to excessive Tau supply in the diet, signalled that excessive Tau may not be conducive to optimal growth in this species.

The gut microbiota community and antioxidant enzymes activity of barramundi reared at seawater and freshwater.

To understand the physiological responses of euryhaline fish to reared salinity, the gut microbiota composition and antioxidant enzymes activity of same batch barramundi Lates calcarifer reared in two extreme salinity condition (seawater and freshwater) were studied, under laboratory condition. The gut bacterial composition was analyzed using high-throughput Illumina sequencing. Acid phosphatase (ACP), catalase (CAT), glutathione peroxidase (GSH-Px), lipid peroxide (LPO), lysozyme (LZM), malonaldehyde (MDA), peroxidase (POD), and superoxide dismutase (SOD) were used as biochemical indicators. The reared salinity did not change the major composition of barramundi gut bacteria under circulating aquaculture system. Proteobacteria, Firmicutes were the most phylum in barramundi gut microbiota community, and Exiguobacterium, Citrobacter, Acinetobacter, Pseudomonas were the dominate genus. CAT and ACP activity in barramundi liver were found significantly different between freshwater and seawater group. GSH-Px, LZM, POD, SOD activity and MDA, LPO levels were not significantly affected by reared salinity. This study is the first high-throughput analyses of the gut microbiota diversity in barramundi from same batch of siblings reared under two extreme salinity condition. And the findings in the present study can be instructive to the management of animal health in barramundi circulating farming activities, and further euryhaline fish gut microecology research.

Beneficial effects of tuna hydrolysate in poultry by-product meal diets on growth, immune response, intestinal health and disease resistance to Vibrio harveyi in juvenile barramundi, Lates calcarifer.

This study was conducted to investigate the effects that tuna hydrolysate (TH) supplementation in poultry by-product meal (PBM) diets would have on growth, immunity and resistance to Vibrio harveyi infection in juvenile barramundi, Lates calcarifer. Five isonitrogenous and isocaloric diets containing fishmeal (FM) without TH supplementation (control) and four diets with 10% TH supplementation viz. a FM protein diet (FMBD + TH), a 75% PBM protein diet (LPBM + TH) and two 90% PBM protein diets, either bioprocessed (BPBM + TH) or unprocessed (HPBM + TH), were formulated for juvenile barramundi, Lates calcarifer. The diets were fed to triplicate groups of juvenile barramundi (average pool weight 12.63 ±â€¯0.11 g) for 10 weeks. Significantly (P < 0.05) higher final body weights and specific growth rates were noted in fish fed with FMBD + TH and BPBM + TH diets when compared to the control. Transmission electron microscopy observation of fish distal intestines revealed a significant enhancement of microvilli length in fish fed FMBD + TH and BPBM + TH whereas scanning electron microscopy analysis found no significant difference in microvilli density. A bacterial challenge with Vibrio harveyi was conducted for 14 days after the growth trial to test the immune response and survival of barramundi. In the pre-challenge condition, a significant reduction in blood glucose was found in BPBM + TH compared to the control, and fish in the post-challenge at 24 h had higher glucose levels compared to fish in the pre- and post-challenge conditions at 72 h. The serum lysozyme activity was significantly higher in FMBD + TH and BPBM + TH compared to the control and fish at 72 h post-challenge exhibited higher lysozyme activity in each treatment compared to all dietary groups in the post-challenge condition at 24 h and to HPBM + TH and BPBM + TH in the pre-challenge condition. Fish fed FMBD + TH, LPBM + TH and BPBM + TH diets had significantly higher survival to the bacterial challenge than fish in the control and HPBM + TH. These results showed that PBM supplemented with TH could successfully replace FM without compromising growth, however, bioprocessed PBM supplemented with TH (BPBM + TH) may significantly improve growth performance, immune response, intestinal health and disease resistance in juvenile barramundi.

Does a bigger mouth make you fatter? Linking intraspecific gape variability to body condition of a tropical predatory fish.

In gape-limited predators, gape size restricts the maximum prey size a predator is capable to ingest. However, studies investigating the energetic consequences of this relationship remain scarce. In this study, we tested the hypothesis that gape-size variability influences individual body condition (a common proxy for fitness) in one of the largest freshwater teleost predators, the barramundi. We found that individual barramundi with larger gapes relative to body size had higher body condition values compared to conspecifics with smaller gapes. Body condition was highest soon after the wet season, a period of high feeding activity on productive inundated floodplains, and body condition decreased as the dry season progressed when fish were restricted to dry season remnant habitats. The increased condition obtained during the wet season apparently offsets weight loss through the dry season, as individuals with large gapes were still in better condition than fish with small gapes in the late-dry season. Elucidation of the links between intraspecific variability in traits and performance is a critical challenge in functional ecology. This study emphasizes that even small intraspecific variability in morphological trait values can potentially affect individual fitness within a species' distribution.

Ulcerative dermatitis in barramundi due to coinfection with Streptococcus iniae and Shewanella algae.

Streptococcus iniae is a pathogenic bacterium which causes septicaemia, while Shewanella algae is an opportunistic pathogen found in marine environments. In this study, we investigated an uncommon coinfection of these 2 bacterial species which resulted in systemic disease and cutaneous ulcers in a barramundi Lates calcarifer farm in the Persian Gulf, Iran. Culture, molecular and histopathological specimens were taken from different organs. In histopathology, results indicated deep bacterial ulceration of skin and subcutaneous muscles. Haemorrhage and hyperaemia were the most common signs observed in visceral organs. In culture, Gram-positive cocci were grown from visceral organs while Gram-negative bacilli were isolated from ulcers. In molecular examination, Streptococcus iniae and Shewanella algae were identified from visceral and ulcer samples, respectively, by PCR of the 16S rRNA gene. The disk diffusion method was used to determine antimicrobial susceptibility of isolated bacteria, with Shewanella algae being resistant to most routinely used antibiotics. In this study, a mixed infection of 2 bacterial species was found; we conclude that systemic streptococcosis could act as a predisposing factor for Shewanella penetration into skin and subsequent ulcer formation. Coinfections are very common in mammals; however, this subject has received little attention in other species, such as fish, and particularly in aquaculture. This study highlights the potential significance of coinfections in barramundi, the effect on the severity of the disease and the potential for new opportunistic pathogens arising.

Impact of dietary starch on extrahepatic tissue lipid metabolism in farmed European (Dicentrarchus labrax) and Asian seabass (Lates calcarifer).

In aquaculture, there is high interest in substituting marine-derived with vegetable-based ingredients as energy source. Farmed carnivorous fish under high carbohydrate diets tend to increase adiposity but it remains unclear if this happens by increased lipid retention/accumulation, promotion of lipogenic pathways, or both. In order to determine the response of extrahepatic tissue to dietary starch, European (Dicentrarchus labrax) and Asian (Lates calcarifer) seabass were fed a control (low starch; LS) or experimental (high starch; HS) diet, for at least 21 days and then transferred for 6 days to saltwater enriched with deuterated water 2H2O. Incorporation of 2H-labelling follows well-defined metabolic steps, and analysis of triacylglycerols (TAG) 2H-enrichment by 2HNMR allowed evaluation of de novo lipogenesis (DNL) in muscle and visceral adipose tissue (VAT). Fractional synthetic rates for TAG-bound fatty acids and glycerol were quantified separately providing a detailed lipogenic profile. The FA profile differed substantially between muscle and VAT in both species, but their lipogenic fluxes revealed even greater differences. In European seabass, HS promoted DNL of TAG-bound FA, in muscle and VAT. High 2H-enrichment also found in muscle TAG-bound glycerol was indicative of its role on lipid cycling. In Asian seabass, HS had no effect on muscle FA composition and lipogenic flux, with no 2H-enriched TAG being detected. VAT on the other hand revealed a strong enhancement of DNL in HS-fed fish along with high TAG-bound glycerol cycling. This study consolidated the use of 2H2O as tracer for fish lipid metabolism in different tissues, under different dietary conditions and suitable to use in different fish models.

Infectious spleen and kidney necrosis disease (ISKND) outbreaks in farmed barramundi (Lates calcarifer) in Vietnam.

Emergence of a disease with clinical signs resembling megalocytivirus infection seriously affected large-scale barramundi farms in Vietnam in 2012-2014 with estimated losses reaching $435,810 per year. An oil-based, inactivated vaccine against red sea bream iridovirus (RSIV) was applied in one farm for disease prevention without analysis of the causative agent, and the farmer reported inadequate protection. Here we describe histological and molecular analysis of the diseased fish. PCR targeting the major capsid protein (MCP) of megalocytiviruses yielded an amplicon with high sequence identity to infectious spleen and kidney necrosis virus (ISKNV) genotype II previously reported from other marine fish but not barramundi. Detection of the virus was confirmed by positive in situ hybridization results with fish tissue lesions of the kidney, liver, pancreas, and brain of the PCR-positive samples. Based on the complete sequence of the MCP gene, the isolate showed 95.2% nucleotide sequence identity and 98.7% amino acid sequence identity (6 residue differences) with the MCP of RSIV. Prediction of antigenic determinants for MCP antigens indicated that the 6 residue differences would result in a significant difference in antigenicity of the two proteins. This was confirmed by automated homology modeling in which structure superimpositioning revealed several unique epitopes in the barramundi isolate. This probably accounted for the low efficiency of the RSIV vaccine when tested by the farmer.

Surveillance for nervous necrosis virus-specific antibodies in barramundi Lates calcarifer in Australian hatcheries.

We conducted single point-in-time and repeated cross-sectional studies of the prevalence of antibodies against nervous necrosis virus (NNV) in populations of adult barramundi Lates calcarifer in Australia. Serum samples collected between 2002 and 2012 were analyzed with indirect ELISA (n = 468). Most of the samples were sourced from broodstock with unknown exposure history, and these were compared with reference populations with confirmed history of exposure to NNV. Non-lethally collected gonad fluid samples from economically valuable barramundi broodstock (n = 164) were tested for the presence of NNV using RT-quantitative PCR at the time of blood sampling to compare infectivity with serostatus, but no virus was detected. NNV-specific immunoreactivity in broodstock was significantly lower than that for immunized and persistently infected populations. Seroprevalence increased over time in broodstock sampled longitudinally, probably reflecting repeated exposure to NNV in a region where the virus was endemic. The seroprevalence for the broodstock was 23.8% over the entire sample period while a cross-sectional survey conducted in 2012 found a seroprevalence of 34.5% with no significant difference between populations based on the geographic region or the history of occurrence of viral nervous necrosis (VNN) disease in the progeny in the respective hatcheries. Although serological surveillance was useful for studying the history of exposure of barramundi to NNV, the lack of association between serostatus in broodstock and the subsequent occurrence of VNN disease in their progeny indicates that ELISA tests for anti-NNV antibodies are not suitable for the purpose of preventing vertical transmission of NNV in barramundi.

Age dependency of nervous necrosis virus infection in barramundi Lates calcarifer (Bloch).

Age-dependent susceptibility to nervous necrosis virus (NNV) was demonstrated for barramundi (Lates calcarifer). The experiment used juvenile barramundi produced from a single spawning that were challenged consecutively by immersion with a redspotted grouper nervous necrosis virus (RGNNV) isolate. The dose and environmental conditions (35 ppt salinity and 30 °C) were constant. Fish and water were sampled longitudinally for histopathology and RT-qPCR analysis to examine the evolution of the disease, virus replication, immune response and release of virus into water. Viral nervous necrosis (VNN) disease occurred in barramundi challenged at 3 and 4 weeks of age while fish challenged at 5, 7 and 9 weeks of age developed subclinical infection. Replication of NNV occurred faster and the concentration of virus reached higher concentrations in the younger fish with clinical disease. Virus isolation and qPCR tests indicated that infectious NNV was released from carcasses into water when fish were affected with clinical disease but not when NNV infection was subclinical. Based on these observations, we consider that carcasses from clinically infected fish have a potentially important role in the horizontal transmission of NNV, and barramundi juveniles should be protected from exposure to NNV until they are 5 weeks of age and reach the disease resistance threshold.

Gelation of barramundi (Lates calcarifer) minced muscle as affected by pressure and thermal treatments at low salt concentration.

BACKGROUND: Barramundi minced muscle with salt 10 g kg-1 and 20 g kg-1 added is gelled by different combinations of pressurisation (300, 400 and 500 MPa at 4 °C for 10 min), cooking (0.1 MPa, 90 °C for 30 min) and setting (0.1 MPa, 50 °C for 2 h) to improve mechanical properties of barramundi gels and reduce salt added to barramundi gels. RESULTS: At the low salt concentration of 10 g kg-1 , pressurisation prior to cooking (P-C) treatment induced barramundi gels with comparable mechanical properties and water-holding capacity to those of conventional heat induced (HI) gels with 20 g kg-1 added salt. At salt concentration of 20 g kg-1 , pressurisation prior to setting (P-S) and P-C gels exhibited higher mechanical properties and water-holding capacity as compared to HI gels. Scanning electron microscopy images showed a smooth and dense microstructure of P-C and P-S gels whereas the microstructure of HI gels is rough and less compact. CONCLUSIONS: P-C treatment can reduce salt concentration added to barramundi gels to 10 g kg-1 . P-S and P-C treatment can result in higher mechanical and functional properties of barramundi gels at conventional salt concentration (20 g kg-1 ) as compared to HI gels. © 2017 Society of Chemical Industry.