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The domestication of a wild-caught aquatic animal is an evolutionary process, which results in genetic discrimination at the genomic level in response to strong artificial selection. Although black tiger shrimp (Penaeus monodon) is one of the most commercially important aquaculture species, a systematic assessment of genetic divergence and structure of wild-caught and domesticated broodstock populations of the species is yet to be documented. Therefore, we used skim sequencing (SkimSeq) based genotyping approach to investigate the genetic structure of 50 broodstock individuals of P. monodon species, collected from five sampling sites (n = 10 in each site) across their distribution in Indo-Pacific regions. The wild-caught P. monodon broodstock population were collected from Malaysia (MS) and Japan (MJ), while domesticated broodstock populations were collected from Madagascar (MMD), Hawaii, HI, USA (MMO), and Thailand (MT). After various filtering process, a total of 194,259 single nucleotide polymorphism (SNP) loci were identified, in which 4983 SNP loci were identified as putatively adaptive by the pcadapt approach. In both datasets, pairwise FST estimates high genetic divergence between wild and domesticated broodstock populations. Consistently, different spatial clustering analyses in both datasets categorized divergent genetic structure into two clusters: (1) wild-caught populations (MS and MJ), and (2) domesticated populations (MMD, MMO and MT). Among 4983 putatively adaptive SNP loci, only 50 loci were observed to be in the coding region. The gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses suggested that non-synonymous mutated genes might be associated with the energy production, metabolic functions, respiration regulation and developmental rates, which likely act to promote adaptation to the strong artificial selection during the domestication process. This study has demonstrated the applicability of SkimSeq in a highly duplicated genome of P. monodon specifically, across a range of genetic backgrounds and geographical distributions, and would be useful for future genetic improvement program of this species in aquaculture.

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Biofloc technology is being applied increasingly in shrimp aquaculture. Many countries have adopted this technology to increase shrimp aquaculture production. Owing to its major importance, this study was conducted to identify the bacterial population from biofloc aquaculture shrimp pond water of Singkir Laut, Kedah, Malaysia. Samples of mature biofloc were collected from the biofloc pond water at different shrimp ages (day of culture [DOC]): DOC30, DOC60 and DOC90. Overall, 43 sequences of bacterial species were blasted, which were identified as Exiguobacterium spp., Bacillus spp., Vibrio spp., Acinetobacter junii, Cobetia marina, Rheinheimera aquimaris and Pseudoalteromonas spp. These bacterial species were identified mostly from heterotrophic plate count agar. The bacterial species of Exiguobacterium aestuarii, Exiguobacterium profundum and Exiguobacterium aurantiacum were identified with 16S rRNA gene sequence similarities of 98.45–99.44%. Bacterial species of Bacillus pumilus, Bacillus velezensis, Bacillus cereus, Bacillus safensis and Bacillus subtilis were identified with 16S rRNA gene sequence similarities of 98.81–99.21%. Bacterial species of Vibrio diazotrophicus, Vibrio diabolicus, Vibrio natriegens and Vibrio harveyi were identified with 16S rRNA gene sequence similarities of 98.03–99.19%. Bacteria from Rheinheimera aquimaris were also identified with 16S rRNA gene sequence similarities of 98.69–99.26%; this bacterium is a novel bacterium identified from biofloc samples of the present study and has not been reported from biofloc samples in any previous study. Exiguobacterium spp. from the novel strains of E. aestuarii and E. profundum were identified as dominant, with 23 of 43 samples isolated from a shrimp pond at time points DOC30, DOC60 and DOC90; these bacteria are Gram‐variable rod‐shaped organisms that are capable of producing extracellular enzyme activity to break down chemical structures, thereby helping to remove organic compounds from aquaculture waste. Exiguobacterium spp. are usually identified in tidal flat sediments and on muddy sea coasts. This discovery is consistent with the site of the shrimp pond near to a muddy beach area, which might be the source of seawater leading to the predominance of this type of bacteria in the biofloc samples in the studied pond.

To assess stress level induced by multiple stressors in aquatic organism, biomarkers have been adopted as early warning indicator due to their high accuracy, rapidity, and sensitivity. We investigated the effects of ectoparasitic isopod infection on heavy metal bioaccumulation (Fe, Cu, Zn, and Cd) in the fish Nemipterus furcosus and profiled the expression of metallothionein (MT) and heat shock proteins 70 (HSP70) genes of the fish host. Sixty individuals (parasitized and nonparasitized with Cymothoa truncata) were collected from three sites differing in the levels of anthropogenic activities off the South China Sea. Our results revealed no significant difference in heavy metal concentrations between infected and nonparasitized fish. We observed a positive correlation between heavy metal bioaccumulation in the fish host and anthropogenic activities. Accordingly, expression analysis of MT genes in fish liver showed significant differences in expression level between sampling sites, with lowest level in the least exploited site (Batu Rakit). A reverse pattern in HSP70 gene expression was demonstrated in fish muscle, showing the highest expression at Batu Rakit. While cymothoid infection in N. furcosus had no significant impact on fish MT gene expression, it resulted in a reduction of HSP70 level in liver of parasitized fish. These findings highlight the putative roles of MT in heavy metal assessment. Future studies should determine the kinetics of cymothoid infection and other potential stressors in characterizing the HSP70 gene expression profile.

Increasing evidence have revealed a positive correlation between gut microbiota and shrimp health, in which a healthy shrimp gut consists of a complex and stable microbial community. Given that both abiotic and biotic factors constantly regulate shrimp gut microbiota, any changes affecting the levels of these factors could cause modification to the gut microbiota assemblage. The goal of this study was to explore the effects of salinity levels and pathogenic Vibrio harveyi infection on the diversity, structure, composition, interspecies interaction, and functional pathways of Litopenaeus vannamei gut microbiota. Juvenile shrimp were cultured at 5 ppt, 20 ppt, and 30 ppt for two months prior to Vibrio harveyi infection. After pathogenic V. harveyi challenge test, genomic DNA was isolated from the shrimp gut, and subjected to the 16S rRNA metagenomic sequencing analysis. We observed that gut microbiota diversity of shrimp cultured at 5 ppt and 30 ppt were lower than those cultured at 20 ppt after exposure to V. harveyi infection, suggesting that shrimp cultured at the two former salinity levels were more susceptible to V. harveyi infection. Network analysis also showed that shrimp cultured at 20 ppt exhibit a more stable bacterial network with complex interspecies interaction, even after induced by V. harveyi. Moreover, the presence of a high number of beneficial bacteria such as Pseudoruegeria, Rhodovulum, Ruegeria, Shimia and Lactobacillus in shrimp cultured at 20 ppt might have played a role in inhibiting the growth of V. harveyi and other potentially pathogenic bacteria. Besides, bacterial functional pathway prediction has also shown that metabolic pathways such as phenylalanine metabolism, glycine, serine and threonine metabolism, starch and sucrose metabolism, glyoxylate and dicarboxylate metabolism, carbon metabolism and biofilm formation process were significantly higher in shrimp cultured at 20 ppt. Collectively, our results suggested that 20 ppt is an optimal salinity that suppresses the growth of V. harveyi and potential pathogenic bacteria in the shrimp gut, which could possibly minimize the risk of pathogenic infection for sustainable production of healthy shrimp.