Size-dependent resistance to amoebic gill disease in naïve Atlantic salmon (Salmo salar).

Amoebic gill disease, caused by the protozoan ectoparasite Neoparamoeba perurans, remains a significant threat to commercial Atlantic salmon aquaculture operations worldwide, despite partial control afforded by selective breeding and therapeutic intervention. Anecdotal reports from commercial producers suggest that historically, smaller Atlantic salmon smolts are more susceptible to AGD than larger smolts. Here, large (>350 g) and small (<200 g) commercially sourced, AGD-naïve Atlantic salmon cohorts were experimentally exposed to 50 N. perurans trophozoites L-1 without intervention. Progression and severity of AGD in challenged cohorts was evaluated through gill pathology, using gill score and histological examination, and quantification of gill-associated amoebae burden using qPCR. To determine the potential basis for differences in AGD susceptibility between cohorts, transcriptome analysis was conducted using RNA extracted from whole gill arches. Overall, the large Atlantic salmon cohort had significantly lower gill parasite burdens and reduced AGD-related gross pathology compared to the small cohort. Relative gill load of N. perurans appeared to be proportional to gill score in both size classes, with larger smolts typically observed to have comparatively reduced parasite burdens at a given gill score. Moreover, comparison between gene expression profiles of large and small smolts highlighted upregulation of genes consistent with elevated immune activity in large smolts. Combined, the results presented here provide strong evidence of size-dependent resistance to AGD in AGD-naïve Atlantic salmon.

Predatory bacteria in the haemolymph of the cultured spiny lobster Panulirus ornatus.

Bdellovibrio and like organisms (BALOs) are Gram-negative obligate predators of other bacteria in a range of environments. The recent discovery of BALOs in the circulatory system of cultured spiny lobster P. ornatus warrants more investigation. We used a combination of co-culture agar and broth assays and transmission electron microscopy to show a Halobacteriovorax sp. strain Hbv preyed upon the model prey bacterium Vibrio sp. strain Vib. The haemolymph microbiome of juvenile P. ornatus was characterised following injection of phosphate buffered saline (control) or prey and/or predator bacteria for 3 d. The predator Hbv had no effect on survival compared to the control after 3 d. However, when compared to the prey only treatment group, lobsters injected with both prey and predator showed significantly lower abundance of genus Vibrio in the haemolymph bacterial community composition. This study indicates that predatory bacteria are not pathogenic and may assist in controlling microbial population growth in the haemolymph of lobsters.

Experimental exposure to low concentrations of Neoparamoeba perurans induces amoebic gill disease in Atlantic salmon.

Amoebic gill disease (AGD) is a significant issue in Atlantic salmon mariculture. Research on the development of treatments or vaccines uses experimental challenges where salmon is exposed to amoebae concentrations ranging from 500 to 5,000/L. However, the water concentrations of N. perurans on affected salmon farms are much lower. The lowest concentration of N. perurans previously reported to cause AGD was 10/L. Here, we report that concentrations as low as 0.1/L of N. perurans can cause AGD. We propose that concentrations of N. perurans that reflect those measured on salmon farms should be used for future experimental challenges.

A highly sensitive, non-invasive qPCR-based strategy for direct quantification of Yersinia ruckeri in fish faeces.

Finfish with asymptomatic Yersinia ruckeri infections pose a major risk as they can transmit the pathogen and cause clinical outbreaks in stock populations. Current tools have insufficient quantitative ability for accurately detecting the trace levels of Y. ruckeri typically associated with asymptomatic infection, necessitate invasive or lethal sampling, or require long processing times. This study presents a highly sensitive qPCR-based method, targeting part of the Y. ruckeri 16S rRNA sequence, that is capable of detecting extremely low levels of Y. ruckeri in noninvasively collected faecal samples. Quantitative precision and accuracy of faecal sample analysis was consistent, despite the complexity of the faecal matrix. The assay demonstrated linearity over a six log-wide dynamic range. Its limit of detection (LOD) and limit of quantification (LOQ) were 4 and 10 copies of the target sequence, respectively. Sensitivity of the assay was comparable to other qPCR-based methods without requiring invasive or lethal sampling. Applicability as a screening strategy was tested using passively collected faecal samples. Asymptomatic Y. ruckeri infection was detected in all samples, although none of the fish exhibited overt infection. This method will be beneficial for finfish disease management if developed further as a noninvasive, screening tool against asymptomatic Y. ruckeri infection.

Culture-independent genomics of a novel chlamydial pathogen of fish provides new insight into host-specific adaptations utilized by these intracellular bacteria.

Several Chlamydiales families are associated with epitheliocystis, a common condition of the fish gill epithelium. These families share common ancestors with the Chlamydiaceae and environmental Chlamydiae. Due to the lack of culture systems, little is known about the biology of these chlamydial fish pathogens. We investigated epitheliocystis in cultured Orange-spotted grouper (Epinephelus coioides) from North Queensland, Australia. Basophilic inclusions were present in the gills of 22/31 fish and the presence of the chlamydial pathogen in the cysts was confirmed by in situ hybridization. Giant grouper (Epinephelus lanceolatus) cultured in the same systems were epitheliocystis free. 16S rRNA gene sequencing revealed a novel member of the Candidatus Parilichlamydiaceae: Ca. Similichlamydia epinephelii. Using metagenomic approaches, we obtained an estimated 68% of the chlamydial genome, revealing that this novel chlamydial pathogen shares a number of key pathogenic hallmarks with the Chlamydiaceae, including an intact Type III Secretion system and several chlamydial virulence factors. This provides additional evidence that these pathogenic mechanisms were acquired early in the evolution of this unique bacterial phylum. The identification and genomic characterization of Ca. S. epinephelii provides new opportunities to study the biology of distantly-related chlamydial pathogens while shining a new light on the evolution of pathogenicity of the Chlamydiaceae.

Thyroid hormone related gene transcription in southern sand flathead (Platycephalus bassensis) is associated with environmental mercury and arsenic exposure.

Arsenic (As) and mercury (Hg) are ubiquitous elements known to disrupt thyroid function in vertebrates. To explore the underlying mechanisms of Hg and As on the fish thyroid system, we investigated the associations between muscle concentrations of Hg and As with thyroid-related gene transcription in flathead (Platycephalus bassensis) from a contaminated estuary. We sampled fish at several sites to determine the hepatic expression of genes including deiodinases (D1 and D2), transthyretin (TTR), thyroid hormone receptors (TRα and TRß) and related them to Hg and As levels in the same individuals. Negative correlations were observed between Hg levels and D2, TTR, TRα and TRß, whereas positive associations were found between As concentrations and TTR and TRß. These results suggest that Hg and As exposures from environmental pollution affect the regulation of genes important for normal thyroid function in fish. These thyroid-related genes could be used as biomarkers for monitoring environmental thyroid-hormone disrupting chemicals.

Immune reactivity in early life stages of sea-cage cultured Pacific bluefin tuna naturally infected with blood flukes from genus Cardicola (Trematoda: Aporocotylidae).

Pacific bluefin tuna (PBT), Thunnus orientalis, due to its high average price on the market is an economically valuable fish species. Infections by blood flukes from the genus Cardicola (Trematoda: Aporocotylidae) represent a growing concern for the cage culture of bluefin tuna in Japan, Australia and Southern Europe. The accumulation of numerous Cardicola eggs in the fish gills causes severe pathology that has been linked to mortality in PBT juveniles up to one year old. The only effective treatment used to mitigate the infection is the oral administration of the antihelminthic drug praziquantel (PZQ) to the affected fish. However, with the need to minimise therapeutic drug use in aquaculture it is hoped that immunoprophylaxis can provide a future alternative to protect the PBT juveniles against Cardicola infection. Currently, little is known of the host immune response to these parasites and of their infection dynamics. In this study, using real-time qPCR we aimed to quantitatively detect C. orientalis and C. opisthorchis DNA within the gills and heart of cultured PBT juveniles and to investigate the host immune response at the transcriptional level in the gills. The research focused mainly during early stages of infection soon after young PBT were transferred to culture cages (from 14 to 77 days post-transfer). An increase (up to 11-fold) of immune-related genes, namely IgM, MHC-I, TCR-ß and IL-1ß was observed in the PBT gills infected with Cardicola spp. (28-77 days post-transfer). Furthermore, IgM (19-fold increase) and MHC-I (11.5-fold increase) transcription was strongly up-regulated in gill samples of PBT infected with C. orientalis relative to uninfected fish but not in fish infected with C. opisthorchis. Cardicola-specific DNA was first detected in the host 14 days post-transfer (DPT) to sea-cages which was 55 days earlier than the first detection of parasite eggs and adults by microscopy. Oral administration of PZQ did not have an immediate effect on parasite DNA presence in the host and the DNA presence started to reduce after 24 days only in the host heart. The results provide evidence of an immune response in early age sea-cage cultured juveniles of PBT naturally infected with C. orientalis and C. opisthorchis. This response, whilst not protective against primary infection, provides evidence that immunisation at an early age may have potential as a health strategy.

Effects of single and repeated infections with Neoparamoeba perurans on antibody levels and immune gene expression in Atlantic salmon (Salmo salar).

Amoebic gill disease (AGD) is the main health problem for the salmon industry in Tasmania, Australia and is now reported in most salmon producing countries. Antibody and gene expression responses to the pathogen, Neoparamoeba perurans, have been studied independently following primary exposure; however, the effects of sequential reinfection, which can often occur during net-pen culture of salmon, remain unclear. The association between the transcription of immunoglobulin (Ig) and their systemic and mucosal antibody levels in regards to AGD is unknown. Herein, we assessed the antibody responses as well as Ig transcription in the gills of Atlantic salmon infected only once and also sequentially with N. perurans. After four successive AGD challenges, no significant differences in plasma or skin mucus levels of IgM were observed between AGD-naïve and challenged fish. However, IgM gene expression in gill lesions of AGD-affected fish increased up to 31 d after infection, while no changes in IgT, TCR and CD8 transcription were observed. Changes at IgM transcription level did not match the lack of antibody response in mucus, which is possibly explained by weak correlations existing between protein and mRNA abundances in cells and tissues. In the second experiment, which investigated Ig responses to AGD at the transcriptional as well as antibody production level in salmon after a single infection, the levels of serum or skin mucus IgM antibody were not affected and no changes in the IgM or IgT transcription were induced.

Vaccination with recombinant protein (r22C03), a putative attachment factor of Neoparamoeba perurans, against AGD in Atlantic salmon (Salmo salar) and implications of a co-infection with Yersinia ruckeri.

Amoebic gill disease (AGD) affects salmonids during the marine grow-out phase in the Tasmanian industry and in other major salmonid producing countries. During the period post-transfer to seawater, the bacterial condition yersiniosis can also cause high levels of mortality in Atlantic salmon grown in Tasmania, in addition to the hatchery outbreaks. The recombinant protein r22C03, a mannose-binding protein-like (MBP-like) similar to attachment factors of other amoebae, was tested as a vaccine candidate against AGD in a large scale challenge trial. Fish were immunised with r22C03 combined with FCA via intraperitoneal (i.p.) injection, and given a booster five weeks later by either i.p. injection (RP group) or by a dip-immersion (mRP). Fish were then challenged twice with Neoparamoeba perurans: the initial challenge 16 weeks after primary immunisation was terminated due to presence of ulcerative lesions in the skin of salmon; the second challenge was carried out after five weeks of treatment with oxytetracycline. These skin lesions might have been associated with a concurrent infection with Yersinia ruckeri, which was detected by real-time qPCR in serum of a large proportion of moribund and survivor fish after the AGD challenge. Before and during the N. perurans infection, levels of antibodies against r22C03 were measured by ELISA in serum, skin mucus and supernatant from skin and gill explants. For the second challenge, the average size of AGD lesions was recorded from histology sections and survival curves were obtained. Before AGD challenge, r22C03 induced antibody responses in serum and explants with both vaccination strategies. At the end of the challenge, levels of antibodies were lower than before challenge irrespective of treatment. Both vaccinated groups presented increased serum antibody responses, while only mRP presented antibody responses in skin mucus, and no significant antibody responses were measured in the explants. Antibodies did not confer protection to N. perurans infection, as no difference was observed in the survival curves of the vaccinated and control groups, and there was no effect on the gill lesion size. The concurrent yersiniosis infection probably represented more closely infection patterns observed in commercial settings. However, it could have interfered with the survival results and with the ability of the fish to respond to the amoebae infection.

Preliminary evidence of transcriptional immunomodulation by praziquantel in bluefin tuna and Atlantic salmon in vitro cultures.

Praziquantel (PZQ), long-used in veterinary and human medicine for the treatment of helminth parasites, is known to enhance humoral and cellular immune responsiveness in mammals but has unknown direct immunomodulatory capabilities in fish. In the present study, we examined the ability of PZQ to induce gene transcriptional changes in immune-competent primary tissue/organ cultures of two highly important yet evolutionarily discrete fish species--Southern bluefin tuna Thunnus maccoyii and Atlantic salmon Salmo salar. These cultures consisted of mixed blood cell population for both species, as well as intestinal explants from bluefin. Although expression profiles varied between species and tissue/organ type, PZQ induced both T-cell receptor (more than twofold) and IL-8 transcriptional expression (more than fourfold). Additionally, increased expression of other inflammatory cytokines including IL-1ß was detected in blood cell cultures from both species, and a general pattern of heightened antiviral signaling was observed. Specifically, elevated transcription of Type I (IFNα) and Type II (IFNγ) interferon in Atlantic salmon blood cultures along with elevated expression of MHC class I in blood cultures of both species. These findings provide preliminary evidence for direct immunomodulation by PZQ in fish and insight into its potential capacity as an immune stimulant/adjuvant in the rapidly expanding aquaculture industry.

Transcriptional immune response of cage-cultured Pacific bluefin tuna during infection by two Cardicola blood fluke species.

Infections by two blood fluke species, Cardicola orientalis and Cardicola opisthorchis, currently present the greatest disease concern for the sea-cage culture of Pacific bluefin tuna (PBT) - a species of high global economic importance and ecological concern. In this study, we aimed to rapidly, quantitatively, and differentially identify infections by these two parasite species in cultured PBT as well as identify potential host immune responses. Using real-time qPCR, we were successful in quantitatively detecting parasite-specific DNA from within host blood, gill, and heart tissues; positively identifying parasitic infections 44 days earlier than microscopy methods previously employed. Both gill and heart became heavily infected by both parasite species in PBT within two months of sea-cage culture, which was only mitigated by the administration of anthelmintic praziquantel. Nevertheless, fish were observed to mount an organ specific transcriptive immune response during infection that mirrored the relative quantity of pathogenic load. In heart, significant (3-6 fold) increases in IgM, MHC2, TCRß, and IL-8 transcription was observed in infected fish relative to uninfected controls; whereas in the gills only IgM transcription was observed to be induced (11 fold) by infection. Interestingly, the relative quantity of IgM transcription was highly correlated to the relative abundance of C. orientalis but not C. opisthorchis DNA in the gill samples, even though this organ showed high prevalence of DNA from both parasite species. Taken together, these findings indicate that although ineffective at combating infection during primary exposure, a cellular immune response is mounted in PBT as a potential rejoinder to future Cardicola exposure, particularly against C. orientalis. Although future investigation into antibody effectiveness will be needed, this work provides valuable preliminary insight into host responsiveness to Cardicola infection as well as additional support for the need of anthelmintic treatment following primary parasite exposure during PBT culture.

Differentially expressed proteins in gill and skin mucus of Atlantic salmon (Salmo salar) affected by amoebic gill disease.

The external surfaces of fish, such as gill and skin, are covered by mucus, which forms a thin interface between the organism and water. Amoebic gill disease (AGD) is a parasitic condition caused by Neoparamoeba perurans that affects salmonids worldwide. This disease induces excessive mucus production in the gills. The host immune response to AGD is not fully understood, and research tools such as genomics and proteomics could be useful in providing further insight. Gill and skin mucus samples were obtained from Atlantic salmon (Salmo salar) which were infected with N. perurans on four successive occasions. NanoLC tandem mass spectrometry (MS/MS) was used to identify proteins in gill and skin mucus of Atlantic salmon affected by AGD. A total of 186 and 322 non-redundant proteins were identified in gill and skin mucus respectively, based on stringent filtration criteria, and statistics demonstrated that 52 gill and 42 skin mucus proteins were differentially expressed in mucus samples from AGD-affected fish. By generating protein-protein interaction networks, some of these proteins formed part of cell to cell signalling and inflammation pathways, such as C-reactive protein, apolipoprotein 1, granulin, cathepsin, angiogenin-1. In addition to proteins that were entirely novel in the context in the host response to N. perurans, our results have confirmed the presence of protein markers in mucus that have been previously predicted on the basis of modified mRNA expression, such as anterior gradient-2 protein, annexin A-1 and complement C3 factor. This first proteomic analysis of AGD-affected salmon provides new information on the effect of AGD on protein composition of gill and skin mucus. Future research should focus on better understanding of the role these components play in the response against infection with N. perurans.

Exploring the pathology of liver, kidney, muscle, and stomach of fledgling seabirds associated with plastic ingestion.

There remain significant gaps in knowledge about 'sub-lethal' impacts of plastic ingestion, particularly chronic impacts on cells, tissues, or organs. Few studies have applied traditional animal health tools, such as histopathology, to assess physiological damage to wildlife, with fewer still providing information on the dosage or exposure to plastics needed to elicit negative effects. Our study seeks to investigate a common hypothesis in plastic pollution research; that an increasing plastics burden will have an impact on an animal's health, examining two wild species with high levels of environmental exposure to plastic through their diet. Here we assess the histopathology of the muscle, upper digestive tract, liver and kidney of two seabird species that are known to be commonly exposed to plastic, comparing exposed and non-exposed individuals. Fledgling seabirds showed histopathological evidence of cumulative pressures such as starvation, disease, and endoparasite burden. However, we observed no evidence of chronic harm that could be explicitly linked to the plastics. We found one case of haemorrhage, reaffirming that large/sharp plastic foreign bodies may cause acute physical damage. Given the numerous interacting pressures on the health of fledging seabirds, including exposure to plastic, this study highlights the need to scrutinise plastic-animal interactions and research though a One Health lens.

Temperature-induced transcription of inflammatory mediators and the influence of Hsp70 following LPS stimulation of southern bluefin tuna peripheral blood leukocytes and kidney homogenates.

Temperature is known to influence inflammatory signalling in mammals, but far less understood in fish. The aim of the present study was to explore the potential effects of temperature on innate immune signalling in head kidney and leukocyte populations of the economically important southern bluefin tuna through the identification and utilization of gene expression targets in vitro. Here, we identified the mRNA sequences of five potential inflammatory mediators - TNFα (1 and 2), IL-1ß, IL-8, and Cox2 - and demonstrate induction of four - TNFα (2), IL-1ß, IL-8, and Cox2 - following LPS stimulation of both peripheral blood leukocytes and head kidney homogenates in vitro by real-time quantitative PCR. Comparison of transcriptional expression in cultures held at 18 and 25 °C (both within the presumed natural temperature range of this heterothermic species) showed accelerated transcription of cytokines TNFα, IL-1ß and IL-8 following LPS stimulation at 25 °C in both tissue types. Peak induction reached comparable levels for each transcript at both temperatures during the 24 h test period with only limited (if any) protraction in expression resulting from cold temperature (18 °C) incubation. Partial mRNA sequences were also identified for both the constitutively expressed and heat inducible chaperone proteins Hsc70 and Hsp70, and 24 h incubation at 25 °C was sufficient to induce Hsp70 transcription in leukocyte but not in head kidney cell populations. Taken together these findings suggest that temperature exerts influence in the timing but not the degree of an innate inflammatory response in bluefin tuna and that different cell populations have differential responsiveness to heat shock in this heterothermic species. Further, LPS stimulation failed to induce Hsp70 at either incubation temperature in leukocytes; whereas 25 °C incubation caused Hsp70 up-regulation in leukocytes with or without the presence of LPS. This suggests that Hsp70 does not play a direct role in immune responsiveness for this species and that an environmental temperature of 25 °C in excess of 24 h initiates a cellular stress response in blood cells of this organism. Lastly, a strong correlation between Hsp70 and IL-8 transcriptional expression was observed following LPS/heat shock stimulation of leukocytes and five potential heat shock response elements were subsequently identified on the gene promoter region of IL-8 indicating that heat shock co-activation of this chemokine previously identified in mammals is also likely present in fish.

Coping with sub-optimal water temperature: modifications in fatty acid profile of barramundi as influenced by dietary lipid.

Metabolic responses to sub-optimal temperature deplete lipid depots, remodel membrane lipid and alter the fatty acid profile in the whole body and tissues of ectothermic vertebrates including fish. The magnitude of these changes may depend on dietary history including oil sources with different fatty acid compositions. Barramundi, Lates calcarifer (Perciformes, Latidae), a tropical ectothermic fish, was fed on diets either rich in dietary long-chain (≥C(20)) polyunsaturated fatty acids (LC-PUFA) from fish oil, rich in stearidonic and γ-linolenic acid (SDA and GLA, respectively) from Echium plantagineum, or rapeseed oil deficient in LC-PUFA. Following 5 weeks at the optimum temperature of 30 °C when growth rates were comparable amongst dietary treatments, water temperature was dropped to 20 °C for 1 week for half of the animals and maintained at 30 °C for the other half. Decreased temperature increased the liver and skeletal muscle content of LC-PUFA in fish fed on echium oil compared with rapeseed oil, while dietary LC-PUFA depots in fish oil fed-fish depleted rapidly in the week of sub-optimal temperature. The lipid unsaturation index of cellular membrane in the liver and muscle increased under low temperature at the same rate regardless of dietary oil. Therefore, rapid exposure of an ectothermic vertebrate to a lower and sub-optimal temperature caused significant modulation in fatty acid composition. We propose that the tolerance of barramundi, a representative of tropical farmed fish, to sub-optimal temperature will be enhanced when fatty acid substrates closer to the LC-PUFA are available in their diet.

Prediction of Feed Efficiency and Performance-Based Traits in Fish via Integration of Multiple Omics and Clinical Covariates.

Fish aquaculture is a rapidly expanding global industry, set to support growing demands for sources of marine protein. Enhancing feed efficiency (FE) in farmed fish is required to reduce production costs and improve sector sustainability. Recognising that organisms are complex systems whose emerging phenotypes are the product of multiple interacting molecular processes, systems-based approaches are expected to deliver new biological insights into FE and growth performance. Here, we establish 14 diverse layers of multi-omics and clinical covariates to assess their capacities to predict FE and associated performance traits in a fish model (Oncorhynchus tshawytscha) and uncover the influential variables. Inter-omic relatedness between the different layers revealed several significant concordances, particularly between datasets originating from similar material/tissue and between blood indicators and some of the proteomic (liver), metabolomic (liver), and microbiomic layers. Single- and multi-layer random forest (RF) regression models showed that integration of all data layers provide greater FE prediction power than any single-layer model alone. Although FE was among the most challenging of the traits we attempted to predict, the mean accuracy of 40 different FE models in terms of root-mean square errors normalized to percentage was 30.4%, supporting RF as a feature selection tool and approach for complex trait prediction. Major contributions to the integrated FE models were derived from layers of proteomic and metabolomic data, with substantial influence also provided by the lipid composition layer. A correlation matrix of the top 27 variables in the models highlighted FE trait-associations with faecal bacteria (Serratia spp.), palmitic and nervonic acid moieties in whole body lipids, levels of free glycerol in muscle, and N-acetylglutamic acid content in liver. In summary, we identified subsets of molecular characteristics for the assessment of commercially relevant performance-based metrics in farmed Chinook salmon.

Comparative transcriptome profiling of virulent and avirulent isolates of Neoparamoeba perurans.

Neoparamoeba perurans, the aetiological agent of amoebic gill disease, remains a persistent threat to Atlantic salmon mariculture operations worldwide. Innovation in methods of AGD control is required yet constrained by a limited understanding of the mechanisms of amoebic gill disease pathogenesis. In the current study, a comparative transcriptome analysis of two N. perurans isolates of contrasting virulence phenotypes is presented using gill-associated, virulent (wild type) isolates, and in vitro cultured, avirulent (clonal) isolates. Differential gene expression analysis identified a total of 21,198 differentially expressed genes between the wild type and clonal isolates, with 5674 of these genes upregulated in wild type N. perurans. Gene set enrichment analysis predicted gene sets enriched in the wild type isolates including, although not limited to, cortical actin cytoskeleton, pseudopodia, phagocytosis, macropinocytic cup, and fatty acid beta-oxidation. Combined, the results from these analyses suggest that upregulated gene expression associated with lipid metabolism, oxidative stress response, protease activity, and cytoskeleton reorganisation is linked to pathogenicity in wild type N. perurans. These findings provide a foundation for future AGD research and the development of novel therapeutic and prophylactic AGD control measures for commercial aquaculture.

Protein metabolism in the liver and white muscle is associated with feed efficiency in Chinook salmon (Oncorhynchus tshawytscha) reared in seawater: Evidence from proteomic analysis.

Understanding the molecular mechanisms that underlie differences in feed efficiency (FE) is an important step toward optimising growth and achieving sustainable salmonid aquaculture. In this study, the liver and white muscle proteomes of feed efficient (EFF) and inefficient (INEFF) Chinook salmon (Oncorhynchus tshawytscha) reared in seawater were investigated by liquid chromatography-tandem mass spectrometry (LC-MS/MS). In total, 2746 liver and 702 white muscle proteins were quantified and compared between 21 EFF and 22 INEFF fish. GSEA showed that gene sets related to protein synthesis were enriched in the liver and white muscle of the EFF group, while conversely, pathways related to protein degradation (amino acid catabolism and proteolysis, respectively) were the most affected processes in the liver and white muscle of INEFF fish. Estimates of individual daily feed intake and share of the meal within tank were significantly higher in the INEFF than the EFF fish showing INEFF fish were likely more dominant during feeding and overfed. Overeating by the INEFF fish was associated with an increase in protein catabolism. This study found that fish with different FE values had expression differences in the gene sets related to protein turnover, and this result supports the hypothesis that protein metabolism plays a role in FE.

Effect of feeding Atlantic salmon (Salmo salar L.) a diet enriched with stearidonic acid from parr to smolt on growth and n-3 long-chain PUFA biosynthesis.

Vegetable oils (VO) have become the predominant substitute for fish oil (FO) in aquafeeds; however, the resultant lower content of n-3 long-chain ( ≥ C20) PUFA (n-3 LC-PUFA) in fish has put their use under scrutiny. The need to investigate new oil sources exists. The present study tested the hypothesis that in Atlantic salmon (Salmo salar L.), a high intake of stearidonic acid (SDA) from Echium oil (EO) would result in increased n-3 LC-PUFA biosynthesis due to a lower requirement for Δ6 desaturase. Comparisons were made with fish fed on diets containing rapeseed oil (RO) and FO in freshwater for 112 d followed by 96 d in seawater. EO fish had higher whole-carcass SDA and eicosatetraenoic acid (ETA) in freshwater and prolonged feeding on the EO diet in seawater resulted in higher SDA, ETA, EPA and docosapentaenoic acid (DPA) compared with RO fish. Fatty acid mass balance of freshwater fish indicated higher biosynthesis of ETA and EPA in EO fish compared with fish fed on the other diets and a twofold increase in n-3 LC-PUFA synthesis compared with RO fish. In seawater, n-3 biosynthetic activity was low, with higher biosynthesis of ETA in EO fish and appearance of all desaturated and elongated products along the n-3 pathway. SDA-enriched VO are more suitable substitutes than conventional VO from a human consumer perspective due to the resulting higher SDA content, higher total n-3 and improved n-3:n-6 ratio obtained in fish, although both VO were not as effective as FO in maintaining EPA and DHA content in Atlantic salmon.

Changes in the Splenic Melanomacrophage Centre Surface Area in Southern Bluefin Tuna (Thunnus maccoyii) Are Associated with Blood Fluke Infections.

Melanomacrophage centres (MMCs) are aggregates of macrophages accumulating various pigments. They have been proposed as an indicator of fish immune response. Blood flukes are common parasites in farmed fish. Two cohorts of wild Southern Bluefin Tuna (Thunnus maccoyi) were examined at transfer, before treatment against blood flukes (pre-treatment) and at harvest. MMCs were assessed in histological sections using image analysis, while Cardicola forsteri and Cardicola orientalis infection severity was determined using qPCR, count of adult flukes in heart flushes and count of eggs in gill filaments. Fish from both cohorts showed the same pattern in the changes in the surface area of MMCs. The surface area of splenic MMCs increased over the ranching duration and was positively correlated to the PCR determined copy numbers of Cardicola forsteri ITS2 rDNA in the gills of those fish. However, the infection with blood fluke was more variable, both between cohorts and individuals within the same cohort. Eggs of blood fluke were detected in renal MMCs using histology. Cardicola forsteri had a higher prevalence than Cardicola orientalis. This study contributes to our understanding of blood fluke infections in Southern Bluefin Tuna and their interactions with MMCs.