Aeromonas salmonicida, a psychrophilic bacterial pathogen, is widely distributed in marine freshwater, causing serious economic losses to major salmon farming areas in the world. At present, it is still one of the most important pathogens threatening salmon farming. Hcp (haemolysin-coregulated protein) is an effector protein in the type-VI secretion system (T6SS), which is secreted by T6SS and functions as its structural component. The results of our previous genomic sequencing showed that hcp existed in the mesophilic A. salmonicida SRW-OG1 isolated from naturally infected Epinephelus coioides. To further explore the role of Hcp in A. salmonicida SRW-OG1, we constructed an hcp-RNAi strain and verified its effect on the virulence of A. salmonicida. The results showed that compared with the wild strain, the hcp-RNAi strain suffered from different degrees of decreased adhesion, growth, biofilm formation, extracellular product secretion, and virulence. It was suggested that hcp may be an important virulence gene of A. salmonicida SRW-OG1.

Vibrio alginolyticus is a common opportunistic pathogen of fish, shrimp, and shellfish, and many diseases it causes can result in severe economic losses in the aquaculture industry. Causing host disease was confirmed by several virulence factors of V. alginolyticus. To date, there have been no reports on the effect of the pstS gene on its virulence regulation of V. alginolyticus. The virulence mechanism of target genes regulating V. alginolyticus is worthy of further study. Previous studies found that Fructus schisandrae (30 mg/mL) inhibited the growth of V. alginolyticus ND-01 (OD₆₀₀ = 0.5) for 4 h, while the expressions of pstS and pstB were significantly affected by F. schisandrae stress. So, we speculated that pstS and pstB might be the virulence genes of V. alginolyticus, which were stably silenced by RNAi to construct the silencing strains pstS-RNAi and pstB-RNAi, respectively. After the expression of pstS or pstB gene was inhibited, the adhesion capacity and biofilm formation of V. alginolyticus were significantly down-regulated. The chemotaxis and biofilm formation ability of pstS-RNAi was reduced by 33.33% and 68.13% compared with the wild-type strain, respectively. Sequence alignment and homology analysis showed that pstS was highly conserved, which suggested that pstS played a vital role in the secretion system of V. alginolyticus. The pstS-RNAi with the highest silencing efficiency was selected for transcriptome sequencing. The Differentially Expressed Genes (DEGs) and GO terms were mapped to the reference genome of V. alginolyticus, including 1055 up-regulated genes and 1134 down-regulated genes. The functions of the DEGs were analyzed by GO and categorized into different enriched functional groups, such as ribosome synthesis, organelles, biosynthesis, pathogenesis, and secretion. These DEGs were then mapped to the reference KEGG pathways of V. alginolyticus and enriched in commonalities in the metabolic, ribosomal, and bacterial secretion pathways. Therefore, pstS and pstB could regulate the bacterial virulence of V. alginolyticus by affecting its adhesion, biofilm formation ability, and motility. Understanding the relationship between the expressions of pstS and pstB with bacterial virulence could provide new perspectives to prevent bacterial diseases.

Pseudomonas plecoglossicida is a common bacterial pathogen in aquaculture, often leading to visceral white spot disease in large yellow croakers (Pseudosciaena crocea). Previous studies have found that certain aptamers show an efficient antibacterial effect against this pathogen. In this study, we analyzed the transcriptome of P. plecoglossicida to get insights into the antibacterial and inhibitions mechanisms following exposure to the aptamer B4. The results showed seven differentially expressed genes (DEGs) associated with the antibacterial effect of the aptamer, namely sad gene encoding aldehyde dehydrogenase, the paaB gene of phenylacetyl coenzyme A cyclooxygenase, the metN1 gene of ABC transporter proteins, two transposase genes with different positions but identical sequences involved in cutting and splicing DNA sequences, and two hypothetical protein genes with unknown functions. Gene Ontology (GO) analysis showed that the DEGs were mainly involved in DNA-mediated translocation, phenylacetic acid catabolism, growth hormone catabolism, polyamine transporter ATPase activity, betaine aldehyde dehydrogenase activity, ABC transporter protein complex, and other related pathways. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the metabolic pathway of niacin and niacinamide mediated through the sad gene was the most significant and relevant, followed by the metabolism of phenylalanine, alanine, aspartic acid and glutamic acid. Real-time quantitative PCR validation showed that the changes in the DEGs were consistent with the transcriptome analysis. These results suggest that the antibacterial aptamer B4 may inhibit P. plecoglossicida by blocking the synthesis of essential nucleic acids and proteins through the modulation of these DEGs and inhibiting their metabolic pathways.