The ocean is a rich resource of flora, fauna, and food. A wild-type bacterial strain showing confluent growth on marine agar with antibacterial activity was isolated from marine water, identified using 16S rDNA sequence analysis as Pseudoalteromonas sp., and designated as strain M2. This strain was found to produce various secondary metabolites including quinolone alkaloids. Using high-resolution mass spectrometry (MS) and nuclear magnetic resonance (NMR) analysis, we identified nine secondary metabolites of 4-hydroxy-2-alkylquinoline (pseudane-III, IV, V, VI, VII, VIII, IX, X, and XI). Additionally, this strain produced two novel, closely related compounds, 2-isopentylqunoline-4-one and 2-(2,3-dimetylbutyl)qunoline-4-(1H)-one, which have not been previously reported from marine bacteria. From the metabolites produced by Pseudoalteromonas sp. M2, 2-(2,3-dimethylbutyl)quinolin-4-one, pseudane-VI, and pseudane-VII inhibited melanin synthesis in Melan-A cells by 23.0%, 28.2%, and 42.7%, respectively, wherein pseudane-VII showed the highest inhibition at 8 µg/mL. The results of this study suggest that liquid chromatography (LC)-MS/MS-based metabolite screening effectively improves the efficiency of novel metabolite discovery. Additionally, these compounds are promising candidates for further bioactivity development.

MAF1 is a global suppressor of RNA polymerase III-dependent transcription, and is conserved from yeast to human. Growing evidence supports the involvement of MAF1 in the immune response of mammals, but its biological functions in fish are unknown. We isolated and characterized Maf1 from the olive flounder Paralichthys olivaceus (PoMaf1). The coding region of PoMaf1 comprised 738 bp encoding a 245-amino-acid protein. The deduced PoMAF1 amino acid sequence shared features with those of MAF1 orthologues from vertebrates. PoMaf1 mRNA was detected in all tissues examined, and the levels were highest in eye and muscle tissue. The PoMaf1 mRNA level increased during early development. In addition, the PoMaf1 transcript level decreased during viral hemorrhagic septicemia virus (VHSV) infection of flounder hirame natural embryo (HINAE) cells. To investigate the role of PoMaf1 in VHSV infection, single-cell-derived PoMaf1 knockout HINAE cells were generated using the clustered regularly interspaced short palindromic repeats/CRISPR-associated-9 (CRISPR/Cas9) system, and cell clones with complete disruption of PoMaf1 were selected. PoMaf1 disruption increased the VHSV glycoprotein (G) mRNA levels during VHSV infection of HINAE cells, implicating PoMAF1 in the immune response to VSHV infection. To our knowledge, this is the first study to characterize fish Maf1, which may play a role in the response to viral infection.

The fish gut microbiota plays an important role in overall health. However, few reports have described the changes in the composition of gut microbiota following infection with pathogenic bacteria in olive flounder (Paralichthys olivaceus). Here, we reported the changes in the gut microbiota composition of flounder after treatment with each of the three pathogenic bacteria (Edwardsiella tarda, Streptococcus iniae, and Vibrio harveyi). Edwardsiella tarda infection decreased the relative abundance of Verrucomicrobia and increased Proteobacteria abundance at the phylum level of the gut microbiota over time. Similarly, Streptococcus iniae infection reduced the relative abundance of Verrucomicrobia. Vibrio harveyi infection caused a decrease in the relative abundance of Firmicutes and Verrucomicrobia and increased Proteobacteria. At the genus level, infection with all three pathogens increased the relative abundance of Ralstonia and Sphingomonas species. Conversely, this infection decreased the relative abundances of Rubritalea, Saccharimonas, and Bacillus species. Therefore, reducing the abundance of Ralstonia and Sphingomonas and increasing the abundance of Rubritalea, Saccharimonas, and Bacillus in the gut microbiota composition of flounder might help maintain a healthy gut microbiota balance. This research might be useful for future studies on improving the health of flounder through gut microbiota regulation.

A Gram-stain-negative, strictly aerobic, non-motile and rod-shaped bacterial strain, MYP1-1^T, was isolated from the intestine of a stalked sea squirt (Styela clava) of the South Sea in the Republic of Korea. The neighbour-joining phylogenetic tree based on 16S rRNA gene sequences revealed that strain MYP1-1^T clustered with the type strains of Halocynthiibacter species and Pseudohalocynthiibacter aestuariivivens . Strain MYP1-1^T exhibited 16S rRNA gene sequence similarity values of 97.0–97.6 % to the type strains of Halocynthiibacter namhaensis , Halocynthiibacter arcticus and P. aestuariivivens . The phylogenetic tree based on genomic sequences showed that strain MYP1-1^T formed a distinct branch separating it from the type strains of two Halocynthiibacter species and P. aestuariivivens and other taxa. The DNA G+C content of strain MYP1-1^T from its genomic sequence was 55.0 mol%. Strain MYP1-1^T contained Q-10 as the predominant ubiquinone and C_18 : 1  ω7c as the major fatty acid. The major polar lipids of strain MYP1-1^T were phosphatidylcholine, phosphatidylglycerol, one unidentified lipid and one unidentified aminolipid. The differences in fatty acid and polar lipid profiles and other differential phenotypic properties made it reasonable to distinguish strain MYP1-1^T from the genera Halocynthiibacter and Pseudohalocynthiibacter . On the basis of the polyphasic taxonomic investigations, we conclude that strain MYP1-1^T constitutes a new genus and species within the class Alphaproteobacteria , for which the name Paenihalocynthiibacter styelae gen. nov., sp. nov. is proposed. The type strain is MYP1-1^T (=KCTC 82143^T=NBRC 114355^T).