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Rhodobacter probiotics are considered as good alternatives to antibiotics for aquaculture. Yet the beneficial effects of Rhodobacter on Chinese mitten crab Eriocheir sinensis are still unclear, and more functions of Rhodobacter supplementation need to be clarified. In this study, a 60-day feeding trial was performed to investigate the protective effects of R. azotoformans against citrobacteriosis in E. sinensis by growth performance, serum immunity, hepatopancreatic antioxidant capability, intestinal flora, and resistance to Citrobacter freundii challenge assays. The results showed that R. azotoformans supplementation significantly and dose-dependently increased weight gain and specific growth rate as well as activities of serum immune and hepatopancreatic antioxidant enzymes, leading to notable improvement in the growth performance, serum immunity and hepatopancreatic antioxidant status of E. sinensis. Besides, R. azotoformans supplementation significantly enhanced intestinal microbial abundance and diversity in E. sinensis, and conferred significant protection of the crabs against C. freundii challenge with seven-day survival rates of 70.0%-100.0%. To the best of our knowledge, this is the first study to reveal the protective effects of R. azotoformans against citrobacteriosis in E. sinensis.

The aim of this study was to evaluate effects of Bacillus licheniformis supplementation on growth, intestine digestive, antioxidant and metabolic capacity, intestine short-chain fatty acids content, and intestine microbiota composition of the Pacific white shrimp Litopenaeus vannamei. In total 2700 shrimp (initial weight 2.5 ± 0.1 g/shrimp) were randomly distributed into six tanks (450 shrimp/500-L tank) and fed at 28 ± 0.5 ℃ for 35 days (four times/day) with control diet and experimental diet supplemented with 108 CFU/g B. licheniformis, followed by an acute nitrite stress for 10 days. At the end of the feeding trial, shrimp dietary B. licheniformis showed improved weight gain, specific growth rate, survival rate after nitrite stress, and decreased feed conversion rate compared to control group. And acetic acid and butyric acid content in the intestine of shrimp dietary B. licheniformis increased significantly except for propionic acid content. In addition, activities of amylase, lipase, and trypsin and activities of glutamine synthetase, hexokinase, and malate dehydrogenase increased significantly compared to control group. After shrimp were exposed to nitrite stress for 10 days, glutathione peroxidase, peroxidase, total antioxidant capacity, and superoxide dismutase levels were still higher in dietary B. licheniformis-shrimp, which showed that B. licheniformis enhanced the resistance to environmental stressor such as nitrite from the level of immune. The 16S rRNA sequencing showed that B. licheniformis increased diversity and abundance of some bacteria. In particular, the abundance of Proteobacteria and Planctomycetes decreased, and the abundance of Firmicutes and Bacteroidetes increased. These results revealed that B. licheniformis could improve the growth performance, increased intestine short-chain fatty acids (SCFA) content, change intestine digestive and metabolic enzyme activity, and regulated microbiota composition, so enhance the intestine antioxidant ability of shrimp subjected to nitrite stress, which is beneficial for shrimp aquaculture.

Providencia rettgeri has been recognized as a zoonotic pathogen of humans and aquaculture animals and has become a global public health concern. However, scarce information is available on the characterization of pathogenic P. rettgeri from kuruma shrimp Marsupenaeus japonicus. In the present study, a P. rettgeri isolate (KM4) was confirmed as a causative agent of red leg disease in cultured M. japonicus, which showed a median lethal dose (LD

Developing ecological approaches for disease control is critical for future sustainable aquaculture development. White spot syndrome (WSS), caused by white spot syndrome virus (WSSV), is the most severe disease in cultured shrimp production. Culturing specific pathogen-free (SPF) broodstock is an effective and widely used strategy for controlling WSS. However, most small-scale farmers, who predominate shrimp aquaculture in developing countries, cannot cultivate SPF shrimp, as they do not have the required infrastructure and skills. Thus, these producers are more vulnerable to WSS outbreaks than industrial farms. Here we developed a shrimp polyculture system that prevents WSS outbreaks by introducing specific fish species. The system is easy to implement and requires no special biosecurity measures. The promotion of this system in China demonstrated that it allowed small-scale farmers to improve their livelihood through shrimp cultivation by controlling WSS outbreaks and increasing the production of ponds., Wang et al. develop a shrimp polyculture system that prevents white spot syndrome outbreaks by introducing specific fish species. The system is easy to implement and requires no special biosecurity measures which makes it ideal for small-scale farmers.

The gut microbiota composition of sea cucumber (Apostichopus japonicas) was investigated using high-throughput sequencing techniques. The mRNA expression of complement component 3 and lysozyme genes was evaluated using quantitative fluorescence PCR. Sea cucumbers were fed with a basal diet (control group) and an experimental diet supplemented with Codonopsis pilosula (experimental group) for 30 days. The results showed that the alpha diversity of the gut microbiota was changed in different indices, including Chao1, the abundance-based coverage estimator, the Shannon index, and Good’s coverage. Dietary C. pilosula promoted the proliferation of the Flavobacteriaceae family of the Proteobacteria phylum and reduced the relative abundance of the Verrucomicrobiaceae family of the Verrucomicrobia phylum. We concluded that dietary C. pilosula supplementation could alter the network interactions among different microbial functional groups by changing the ecological network’s microbial community composition and biological evolution. A positive effect on A. japonicus immune responses in the gut was seen via increasing the mRNA expression of the complement component 3 and lysozyme genes. It seems to happen via modulating the balance in gut microbiota.

The genus Acinetobacter has recently emerged as bacterial pathogens associated with mass mortalities in aquaculture. Yet scarce information is available on Acinetobacter venetianus as a potential pathogen of whiteleg shrimp Penaeus vannamei. In the present study, an A. venetianus pathogen (SHF1) was confirmed as a causative agent of red leg disease in freshwater-cultured whiteleg shrimp. It showed a mean lethal dose (LD50) value of 3.8 × 105 CFU mL−1 and developed multiple resistances to phenicols, quinolones and tetracycline drugs used in aquaculture. In addition, the pathogenic A. venetianus could be effectively controlled by Bdellovibrio powder at 0.8 mg L−1, which exhibited significant antibacterial effect and displayed good protection against A. venetianus challenge in P. vannamei for six days. As far as we know, this is the first study to identify A. venetianus as a potential bacterial pathogen of red leg disease in freshwater-farmed whiteleg shrimp.

Fat greenling (Hexagrammos otakii) is an important commercial marine fish species cultured in northeast Asia, but its available gene sequences are limited. Vibrio harveyi is a causative agent of vibriosis in fat greenling and also causes severe losses to the aquaculture industry in China. In order to obtain more high-quality transcript information and investigate the early immune response of fat greenling against V. harveyi, the fish were artificially infected with V. harveyi, and five sampling points were set within 48 h. Iso-Seq combined with RNA-Seq were applied in the comprehensive transcriptome analysis of V. harveyi-infected fat greenling. Total 42,225 consensus isoforms were successfully extracted from the result of Iso-Seq, and more than 19,000 ORFs were predicted. In addition, total three modules were identified by WGCNA which significantly positive correlated to the infection time, and the KEGG analysis showed that the immune-related genes in these modules mainly enriched in TLR signaling pathway, NF-κB signaling pathway and Endocytosis. The activation of inflammation and endocytosis was the most significant characteristics of fat greenling immune response during the early infection. Based on the WGCNA, a series of high–degree nodes in the networks were identified as hub genes. The protein structures of cold-inducible RNA-binding protein (CIRBP), poly [ADP-ribose] polymerase 1 (PARP1) and protein arginine N-methyl transferase 1 (PRMT1) were subsequently found to be highly conserved in vertebrate, and the gene expression pattern of CIRBP, PARP1, PRMT1 and a part of TLR/NF-κB pathway-related genes indicated that these proteins might have similar biological functions in regulation of inflammatory response in teleost fish. The results of this study provided the first systematical full-length transcriptome profile of fat greenling and characterized its immune responses in early infection of V. harvey, which will serve as the foundation for further exploring the molecular mechanism of immune defense against bacterial infection in fat greenling.

Aeromonas salmonicida subsp. masoucida (ASM) is an important bacterial pathogen of salmonid fish, which can cause huge economic losses to the fish farming industry. In order to screen effective vaccine candidate proteins, four outer membrane proteins of ASM, including OmpA, OmpC, OmpK and OmpW, were selected and recombinantly expressed in Escherichia coli. The result of western blotting showed that these four recombinant proteins could be recognized by rainbow trout anti-ASM antibodies. The immune protective effects of the four rOMPs were also investigated, and the relative percentage survival (RPS) of rOmpA, rOmpC, rOmpK and rOmpW were 71.1%, 81.6%, 55.3% and 42.1%, respectively. The RPS of rOmpC was significantly higher than the other three rOMPs, so the immune responses of rainbow trout induced by rOmpC were further investigated. The results showed that vaccination with rOmpC could significantly induced the production of specific serum antibodies and proliferation of sIg + lymphocytes in peripheral blood. Meanwhile, RT-qPCR analysis showed that rOmpC could significantly enhance the expression of the MHC-II, TCR, CD4, CD8, IL-8 and IgM genes compared with the BSA immunized group. These results demonstrated that rOmpC could induce strong humoral immune response in rainbow trout and provided effective immune protection against ASM challenge, which indicated that OmpC is a promising vaccine candidate against Aeromonas salmonicida infection.

Fat greenling (Hexagrammos otakii) is an important aquaculture fish species in northern China. Unfortunately, Vibrio infections have caused considerable losses to the fat greenling aquaculture industry. However, the study on immune response of fat greenling against Vibrio species has not been reported yet. In this paper, the immune response of fat greenling against V. harveyi at gene expression level was studied by transcriptome analysis. A total of 189753 high-quality unigenes with a N50 length of 672bp were obtained by transcriptome profiling, which provided abundant data for the future study of fat greenling. Comparative analysis showed that 5425 differentially expressed genes (DEGs) were identified on day 3 post-infection (3dpi), containing 1837 up-regulated and 3588 down-regulated genes. Further annotation and analysis revealed that the DEGs were enriched in complement and coagulation cascades, ribosome, oxidative phosphorylation, glycine, serine and threonine metabolism and peroxisome proliferator-activated receptor (PPAR) signaling pathway. These pathways were mainly associated with phagocytosis and pathogen clearance, rarely involved in bacteria adhesion and pathogen identification, which suggested that the host might begin to clear and kill the invading bacteria on 3dpi. The research might provide a valuable resource to further study immune response and suggest strategies against V. harveyi infection in fat greenling.

Grading procedure in routine sea cucumber hatchery production is thought to affect juvenile sea cucumber immunological response. The present study investigated the impact of a 3-min mechanical perturbation mimicking the grading procedure on neuroendocrine and immune parameters of the sea cucumber Apostichopus japonicus. During the application of stress, concentrations of noradrenaline and dopamine in coelomic fluid increased significantly, indicating that the mechanical perturbation resulted in a transient state of stress in sea cucumbers. Coelomocytes concentration in coelomic fluid increased transiently after the beginning of stressing, and reached the maximum in 1 h. Whereas, coelomocytes phagocytosis at 3 min, superoxide anion production from 3 min to 0.5 h, acid phosphatase activity at 0.5 h, and phenoloxidase activity from 3 min to 0.5 h were all significantly down-regulated. All of the immune parameters recovered to baseline levels after the experiment was conducted for 8 h, and an immunostimulation occurred after the stress considering the phagocytosis and acid phosphatase activity. The results suggested that, as in other marine invertebrates, neuroendocrine/immune connections exist in sea cucumber A. japonicus. Mechanical stress can elicit a profound influence on sea cucumber neuroendocrine system. Neuroendocrine messengers act in turn to modulate the immunity functions. Therefore, these effects should be considered for developing better husbandry procedures.

We report the effect of transport stress on the immune responses of the sea cucumber Apostichopus japonicus. The total coelomocytes counts, phagocytosis activity, reactive oxygen species production, acid phosphatase activity, phenoloxidase activity and catecholamine levels were measured in A. japonicus following both damp and water transportation methods. With both transport methods, stress resulted in significant elevation of catecholamine levels. Total coelomocytes counts increased following transportation, whereas phagocytic activity and reactive oxygen species production decreased. Damp transport significantly depressed acid phosphatase and phenoloxidase activity in A. japonicus. However, there were no significant differences in acid phosphatase and phenoloxidase activity after water transportation. These results show that transport stress induces modifications in the immune system of A. japonicus and that damp transport elicited more severe stress than water transport. This study increased understanding of the mechanism of transport stress inducing neuroendocrine responses and immune modification in A. japonicus. The information obtained should be helpful to optimize transport process in sea cucumber culture.

Bacteriosis is a major economic problem affecting the farming industry of whiteleg shrimp Penaeus vannamei. Little information is available on Providencia rettgeri as a possible pathogen for P. vannamei. In this study, a virulent strain, here named S6, was isolated from diseased P. vannamei, identified phenotypically, and molecularly, as P. rettgeri. A phylogenetic tree was constructed to examine the taxonomic position and relatedness of isolate S6 to other P. rettgeri isolates. When screened against a range of common veterinary antibiotics, isolate S6 exhibited resistance to aminoglycosides and sulfonamides antibiotics in aquaculture. To the best of our knowledge, this is the first report of P. rettgeri as a causal agent of bacteriosis in freshwater farmed P. vannamei.

Tail rot disease is the cause of significant economic damage in freshwater farmed Murray cod Maccullochella peelii. Only scarce information is available on Aeromonas hydrophila as a possible causal agent for this disease. In this study, a virulent strain, temporarily named XY3, was isolated from diseased codfish suffering from tail rot disease, and identified as A. hydrophila through phylogenetic analysis and phenotypic characteristics. A. hydrophila possesses multiple virulence genes including aerA, ahpA, alt, ast and hlyA genes. In addition, it appears that isolate XY3 has developed multiple resistances to cephalosporin, chloromycetin, glycopeptides, macrolides, nitrofuran, and penicillin drugs, as well as to aminoglycosides, sulfonamides, and tetracyclines antibiotics for veterinary uses in aquaculture as revealed when screened against a range of common antibiotics. To the best of our knowledge, this is the first report of tail rot disease caused by A. hydrophila in freshwater farmed codfish.