Bacterial Flora in the Gill Tissues and Intestinal Tracts of Male and Female Chinese Mitten Crabs (Eriocheir sinensis) with Different Diets in a Mud Pond.

The Chinese mitten crab, Eriocheir sinensis, is an economically valuable aquaculture species. Prior to sale, farmed crabs are often fattened with pellet feed or wild fish. In this study, PacBio Sequel sequencing was used to determine the bacterial flora in the intestinal tracts and gill tissues of male and female E. sinensis fed with various diets. The flora was then compared with the microorganisms found in environmental samples. The results showed that Proteobacteria was the dominant phylum in both tissue and environmental samples. The relative abundances of Proteobacteria in the water grass surface flushing samples and water grass samples were the highest, at up to 95.68% and 67.85%, respectively. Beyond that, Bacteroidetes, Firmicutes, and Tenericutes were the dominant phyla (>1%) in the intestinal samples, whereas Bacteroidetes and Actinobacteria were the dominant phyla in the gills. In addition, different environment samples contained diverse bacterial phyla, indicating some differences in the community composition between the different sample groups. Heat map clustering and principal coordinate plot analyses indicated that intestinal samples, crab gill samples, and environmental samples clustered together, respectively. Furthermore, an unweighted pair-group method with arithmetic mean technique confirmed that the intestinal and gill samples of crabs with different diets separately clustered together, suggesting the microbial assemblages of the same tissues share a greater similarity than those from crabs of different sex and eating different diets. What's more, biomarker bacteria (LDA ≥ 4) from the different groups were identified. Pathogenic agents from the genus Aeromonas were abundant in the intestinal samples of crabs fed with pellet feed, and Vibrio species were prevalent in the intestinal samples of crabs fed with wild fishes.

Effects of different salinity on the transcriptome and antibiotic resistance of two Vibrio parahaemolyticus strains isolated from Penaeus vannamei cultured in seawater and freshwater ponds.

The transcriptome and antibiotic resistance of Vibrio parahaemolyticus isolated from Penaeus vannamei cultured in seawater (strain HN1)and freshwater (strain SH1) ponds were studied at different salinity (2‰ and 20‰). At different salinity, 623 differentially expressed genes (DEGs) significantly upregulated and 1,559 DEGs significantly downregulated in SH1. In HN1, 466 DEGs significantly upregulated and 1,930 DEGs significantly downregulated, indicating high salinity can lead to the downregulation of most genes. In KEGG analysis, the expression of DEGs annotated to starch and sucrose metabolism pathway was higher at 2‰ salinity than at 20‰ salinity in HN1 and SH1, implying salinity affected bacterial growth mainly through this pathway. In the enrichment analysis of upregulated DEGs, two pathways (Valine, leucine, and isoleucine degradation, and Butanoate metabolism) were significantly enriched at different salinity. Antibiotic-susceptibility test discovered that SH1 isolated from P. vannamei cultured in freshwater was resistant to multiple drugs, including kanamycin, gentamicin, medemycin, and azithromycin, at a salinity of 2‰, whereas at 20‰ salinity, SH1 was not resistant to the drugs. The HN1 strain isolated from P. vannamei cultured in mariculture was resistant to polymyxin B and clindamycin at 20‰ salinity. Whereas, HN1 was intermediately susceptible to these two antibiotics at 2‰ salinity. These results indicate that the drug resistance of bacteria was affected by salinity. Furthermore, beta-lactam resistance was significantly enriched in SH1 at different salinity, and the inhibition zone of penicillin G was consistent with the results of a beta-lactam resistance pathway.

The composition of the microbial community associated with Macrobrachium rosenbergii zoeae varies throughout larval development.

The giant river prawn, Macrobrachium rosenbergii, is an economically important freshwater prawn. The cultivation of zoea larvae is crucial for the success of the M. rosenbergii industry. In this study, we surveyed the microbial community diversity and structure associated with M. rosenbergii zoeae at different stages of larval development. Samples of zoea larvae from different developmental stages were collected and subjected to high-throughput DNA sequencing. Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria were the dominant phyla in all six sample groups. At the genus level, the relative abundance of Bacillus decreased, and that of Enterobacter increased with the growth of the zoeae. This may have been related to the intestinal development of the zoea larvae. The microbial diversity of M. rosenbergii zoea larvae decreased significantly with development. The beta diversity analysis showed that the closer the developmental stage of M. rosenbergii, the more similar the structure of the associated bacterial communities.

A modification of nested PCR method for detection of Enterocytozoon hepatopenaei (EHP) in giant freshwater prawn Macrobrachium rosenbergii.

The microsporidian Enterocytozoon hepatopenaei (EHP) has become a critical threat to the global shrimp aquaculture industry, thus necessitating early detection by screening. Development of a rapid and accurate assay is crucial both for the active surveillance and for the assessment of shrimp with EHP infection. In the present study, a distinct strain of E. hepatopenaei (EHP Mr ) was found in Macrobrachium rosenbergii. The SWP1 gene analysis revealed it was a new genotype that differed with the common strain isolated from the Litopenaeus vannamei (EHP Lv ). A nested SWP-PCR method was modified to fix the bug that the original inner primers could not recognize the EHP Mr strain. The redesigned inner primers successfully amplified a product of 182 bp for both the EHP Mr strain and the EHP Lv strain. The new primers also had good specificity and high sensitivity, which may serve as an alternative for EHP genotyping. This study provided a method for detection of EHP in the biosecurity of Macrobrachium rosenbergii farming, and the developed protocol was proposed for the routine investigation and potential carrier screening, especially for molecular epidemiology.