Cryopreservation of spermatogonia could be a useful tool to preserve the genetic resources of fish, which could be further restored via germ cell transplantation. In this study, the protocol for the cryopreservation of the spermatogonia of Asian sea bass (Lates calcarifer), an economically important fishery resource in the Indo-West Pacific, was optimised. The impact of the cryopreservation technique on cell viability and apoptosis, expression of several genes related to immature germ cell markers, transplantability in allogeneic recipients, and global DNA methylation was evaluated. The slow-freezing method was performed for the cryopreservation of immature testis tissue, which contains a high proportion of spermatogonia. The optimal condition that yielded the highest recovery rate of post-thawed spermatogonia included a cryomedium containing Leibovitz\'s (L-15) medium and 10 % dimethyl sulfoxide, ice equilibration for 60 min before freezing, and subsequent thawing at 4 °C for 8 min. Moreover, a higher number of early and late apoptotic cells was detected in the cryopreserved than in the fresh testes, suggesting that apoptosis could result in reduced viability. The expression levels of dazl decreased in the cryopreserved testes; however, there were no significant differences in the expression levels of nanos2 or nanos3 between the fresh and cryopreserved testes. Although qRT-PCR showed lower vasa expression in cryopreserved testicular cells, in situ hybridisation showed expressed vasa in the cryopreserved testicular cells. Post-thawed spermatogonia could be incorporated into the genital ridge of allogeneic recipients, suggesting that cryopreserved spermatogonia exhibit transplantability characteristics. Compared with fresh testes, significant changes in the proportion of DNA methylation (decreased 5-mC and 5-caC) were observed in cryomedium-free testicular cells, whereas those of the cryopreserved cells were not significantly different. Therefore, the method we developed for the cryopreservation of the spermatogonia of Asian sea bass enabled post-thaw cells to retain several stemness characteristics and maintain their epigenetic stability.

Advancements in genome sequencing and assembly techniques have increased the documentation of structural variants in wild organisms. Of these variants, chromosomal inversions are especially prominent due to their large size and active recombination suppression between alternative homokaryotypes. This suppression enables the 2 forms of the inversion to be maintained and allows the preservation of locally adapted alleles. The Barramundi Perch (BP; Lates calcarifer) is a widespread species complex with 3 main genetic lineages located in the biogeographic regions of Australia and New Guinea (AUS + NG), Southeast Asia (SEA), and the Indian Subcontinent (IND). BP are typically considered to be a protandrous sequential hermaphrodite species that exhibits catadromy. Freshwater occupancy and intraspecific variation in life history (e.g. partially migratory populations) exist and provide opportunities for strongly divergent selection associated with, for example, salinity tolerance, swimming ability, and marine dispersal. Herein, we utilize genomic data generated from all 3 genetic lineages to identify and describe 3 polymorphic candidate chromosomal inversions. These candidate chromosomal inversions appear to be fixed for ancestral variants in the IND lineage and for inverted versions in the AUS + NG lineage and exhibit variation in all 3 inversions in the SEA lineage. BP have a diverse portfolio of life history options that includes migratory strategy as well as sexual system (i.e. hermaphroditism and gonochorism). We propose that the some of the life history variabilities observed in BP may be linked to inversions and, in doing so, we present genetic data that might be useful in enhancing aquaculture production and population management.

The advent of aquaculture has been one of the most significant shifts in world food supply during the last century. Aquaculture has rapidly expanded and become a global food industry, spurred by population expansion, increased seafood consumption, and decreased captured fisheries. Nonetheless, the exponential growth of aquaculture has emerged as a significant contributor to anthropogenic changes. Unexpectedly, the result has focused in the emergence and spread of new diseases. The Asian sea bass (Lates calcarifer) is an economically important species in aquaculture, contributing significantly to the global seafood market. However, bacterial diseases have emerged as a major concern, affecting both wild and cultured populations of this species. The most prevalent bacterial pathogens are streptococcus, vibriosis, nocardiosis, tenacibaculosis, and pot-belly disease. Therefore, this review aims to comprehensively analyze both emerging and non-emerging bacterial diseases affecting L. calcarifer and explore potential management approaches for their control. Through an extensive literature survey and critical evaluation of research findings, this review highlights the current understanding of bacterial diseases in L. calcarifer and proposes strategies for better disease management. In addition, this review looks at the rise and characteristics of aquaculture, the major bacterial pathogens of L. calcarifer and their effects, and the specific attributes of disease emergence in an aquatic rather than terrestrial context. It also considers the potential for future disease emergence in L. calcarifer due to aquaculture expansion and climate changes.

In 2015 and 2016, two Barramundi (Lates calcarifer) farms in Singapore reported a disease outbreak characterized by lethargic behavior, pronounced inappetence, generalized skin lesions, erosions of the fins and tail, and ultimately high mortality in their fish. Next-generation sequencing and PCR confirmed presence of a novel virus belonging to the Alloherpesviridae family, Lates calcarifer herpesvirus (LCHV), which was subsequently isolated and cultured. We characterize, for the first time, the complete genome of two cultured LCHV isolates. The genome contains a long unique region of approximately 105,000 bp flanked by terminal repeats of approximately 24,800 bp, of which the first 8.2 kb do not show any similarity to described genomes in the Alloherpesviridae family. The two cultured isolates share 89% nucleotide identity, and their closest relatives are the viruses belonging to the genus Ictalurivirus. Experimental infections using one of the cultured LCHV isolates resulted in identical clinical signs as originally described in the index farm, both in intraperitoneal-injection infected fish and cohabitant fish, with mortality in both groups. Histopathological analysis showed pronounced abnormalities in the gills. Virus culture and PCR analysis confirmed the replication of LCHV in the infected fish, and thus Koch\'s postulates were fulfilled.

Members of the genus Erysipelothrix are emergent pathogens of cultured eels, as well as several characid and cyprinid species. Since 2013, E. rhusiopathiae has been reported from diseased barramundi (Lates calcarifer) cultured in North America; we recovered 8 E. rhusiopathiae isolates from diseased fish during different outbreaks from the same farm. The E. rhusiopathiae isolates from barramundi were compared phenotypically and genetically to E. piscisicarius isolates characterized from ornamental fish and E. rhusiopathiae recovered from aquatic and terrestrial animals. All barramundi isolates were PCR-positive for the surface protective antigen type B (spaB) gene, and shared ≥ 99.7% sequence similarity among concatenated multilocus sequence analysis gene sequences, indicating a high degree of genetic homogeneity. These isolates were > 99% similar to other spaB-positive isolates from marine invertebrates and marine mammals, consistent with findings for other spa types. The spaA and spaB isolates shared < 98% similarity, as well as < 90% similarity with spaC-positive E. piscisicarius. Similar clonality among the spaB isolates was observed using repetitive element palindromic PCR. In experimental intracoelomic injection challenges conducted to fulfill Koch postulates, 67% of exposed tiger barbs (Puntigrus tetrazona) died within 14 d of challenge. Our study supports previous work citing the genetic variability of Erysipelothrix spp. spa types and the emergence of members of the genus Erysipelothrix as nascent fish pathogens.

Global warming is leading to an increase in the frequency and intensity of extreme weather events, magnifying the breadth of temperatures faced by ectotherms across days and seasons. Despite the importance and ecological relevance of diurnal thermal variability, the vast majority of knowledge on gene expression patterns and physiology stems from animals acclimated to constant temperatures or in the early stages of exposure to a new temperature regime. If heterothermal environments modulate responses differently from constant thermal environments, our existing capacity to forecast impacts of climate warming may be compromised. To address this knowledge gap, we acclimated barramundi (Lates calcarifer) to 23 °C, 29 °C (optimal), 35 °C and to thermal cycling conditions (23-35 °C daily with a mean of 29 °C) and sampled liver and white muscle tissue before acclimation and after 2 and 17 weeks of acclimation. NanoString nCounter technologies were used to measure expression of 20 genes related to metabolism, growth and maintenance of cellular homeostasis. Acclimation to cool and warm conditions caused predictable changes in whole-animal performance (metabolism and growth) and the underlying gene expression patterns. Acclimation to a cycling temperature regime did not change the molecular regulation of metabolism or growth compared with barramundi acclimated to constant 29 °C, nor did it cause any discernible effects on whole-animal performance. However, the heat shock response was higher in the former group, suggesting that barramundi under a daily temperature cycle have an increased need for cellular chaperoning to minimise detrimental effects of temperature on proteins. We conclude that the genetic regulation of metabolism and growth may be more dependent on the mean daily temperature than on the daily temperature range.

Microwave-assisted induction heating (MAIH) is a composite microwave and induction heating to supply rapid and uniform heating of food. A recent study showed that the optimum MAIH heating condition for barramundi meat was 90 °C/110 s or 70 °C/130 s. This study examines whether the microwave-assisted induction heating (MAIH) technology (at 70 °C for 130 s or 90 °C for 110 s) can more effectively slow down the quality loss of barramundi meat during cold storage than the traditional boiling method (at 90 °C for 150 s). The results show that no microbial growth was observed in the three groups of heated barramundi meat samples during the 60 days of cold storage. However, the MAIH technology slowed down the increase in the total volatile basic nitrogen (TVBN) content more significantly than the boiling method. As the cold storage time increased, though, the L* (lightness), a* (redness), and W (whiteness) values decreased, while the b* (yellowness) and color difference (ΔE) values increased in the three treatment groups. However, the MAIH technology slowed down the decrease in the L*, a*, and W values more significantly, and produced a ΔE value smaller than the boiling method. Moreover, the MAIH technology ensured higher hardness and chewiness of the barramundi meat than the boiling method. Overall, the MAIH technology slowed down the quality loss of the barramundi meat and maintained better color and texture during cold storage.

AbstractGiven the critical role of metabolism in the life history of all organisms, there is particular interest in understanding the relationship between individual metabolic phenotypes and the capacity to partition energy into competing life history traits. Such relationships could be predictive of individual phenotypic performances throughout life. Here, we were specifically interested in whether an individual fish\'s metabolic phenotype can shape its propensity to feed following a significant stressor (2-min exhaustive exercise challenge). Such a relationship would provide insight into previous intraspecific observations linking high metabolism with faster growth. Using a teleost fish, the barramundi (Lates calcarifer), we predicted that individuals with high standard metabolic rates (SMRs) and maximal metabolic rates (MMRs) would be faster to recover and resume feeding after exercise. Contrary to our prediction, neither SMR nor MMR was correlated with latency to feed after exercise (food was offered at 0.5, 1.5, 3, and 18 h after exercise). Only time after exercise and individual fish ID were significant predictors of latency to feed. Measurements of MMR from the same individuals (three measurements spaced 8-12 d apart) revealed a moderate degree of repeatability (R=0.319). We propose that interindividual differences in biochemical and endocrine processes may be more influential than whole-organism metabolic phenotype in mediating feeding latency after exercise.

Drug behavior in the bodies of fish is largely influenced by the water temperature. Antimicrobial drugs are needed for the control of bacterial outbreaks in farmed fish including Asian seabass (Lates calcarifer). However, little is known about the temperature effect on appropriate drug uses in this species. The purpose of this study was to investigate the differences in pharmacokinetics (PK), optimal dosages, tissue depletion, and withdrawal time (WDT) of florfenicol (FF) in Asian seabass reared at 25 and 30 °C. In the PK study, the fish were administered with a single oral dose of 10 mg/kg FF. The optimal dosing regimen was determined by the pharmacokinetic-pharmacodynamic (PK-PD) approach. In the tissue depletion and WDT study, FF was administered at the optimal dosages once daily for 5 days and the WDT was determined by linear regression analysis based on the sum of FF and its metabolite florfenicol amine (FFA) in the muscle/skin. When the temperature was increased from 25 to 30 °C, the elimination half-life of FF was significantly decreased from 11.0 to 7.2 h. While the other PK parameters were not changed significantly, the calculated optimal dosages for the target minimum inhibitory concentration (MIC) of 2 µg/mL were 10.9 and 22.0 mg/kg/day, respectively for 25 and 30 °C. The sum of FF + FFA is a preferable marker residue for WDT determination because differential FF metabolism was observed at different temperatures. The depletion half-life of the muscle/skin was shortened from 41.1 to 32.4 h by the 5 °C temperature increase. Despite different absolute amounts of FF given between the two temperature levels, the WDTs were very similar at 6-7 days. Thus, it appears that a single temperature-independent WDT can potentially be assigned when the drug was applied at the optimal dosage.

The citric acid (CA) and electrolysed water (EW) are considered effectively in inactivating microorganisms. The objective of this study was to explore the bactericidal mechanism of CA combined with EW on Aeromonas spp. in barramundi (Lates calcarifer) by in vitro metabolomics method. This study determined the survival population of three strains of Aeromonas bacteria (strain 1: Aeromonas salmonicida strain A1 (skin); strain 2: A. veronii strain Til2 (gut), and strain 3: A. hydrophila strain B11 (gill)), which were isolated and identified from putrid barramundi treated alone or in combination with 1 % CA and EW (free available chlorine (FAC) 25 mg/L, pH 3.23, oxidation-reduction potential (ORP) 1015 mV). The bactericidal mechanism was investigated by microbiological analysis, nuclear magnetic resonance (NMR), multivariate data analysis, and fluorescence staining analysis. The results showed that the combined treatment significantly reduced the number of Aeromonas bacteria at 1.64-1.69 log CFU/g and extended the shelf life of barramundi fillets. In addition, the combined treatment had a higher effect on the cell membrane integrity of the bacteria. In total, 36 metabolites were identified in the three strains. The undissociated molecules of CA can enter the cytoplasm, resulting in cell damage and inhibiting metabolic pathways. EW could lead to the reduction of metabolic products caused by oxidative stress and acid stress. Under the synergistic stress of CA and EW, the changes of main metabolite contents in the combined treatment group were significantly reduced. After combined treatment, there were 20, 31, and 31 pathways in which carbohydrate metabolism, amino acid metabolism, and energy metabolism were changed considerably. These findings indicated that the bactericidal mechanism of the bactericidal substance might be explained by the interference of the metabolic pathway, which guided post-treatment sanitisation and extended the applicability of the NMR spectrum to specific spoilage organisms (SSO) analysis in fish.