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Although members of the genus Acinetobacter have emerged as important nosocomial pathogens causing severe human infections, there are few reports about their occurrence as fish pathogens. In this study, five bacterial strains were isolated from diseased loach (Misgurnus anguillicaudatus) cultured in a farm in China. The diseased loach displayed shedding of skin mucus and many petechial haemorrhages all over the body. Based on sequence analyses of 16S rRNA and rpoB genes, the isolates were identified as Acinetobacter pittii. An experimental infection assay confirmed their pathogenicity to loach. The results of artificial infection in zebrafish (Barchydanio rerio) and nematode (Caenorhabditis elegans) suggested that, as well as loach, these A. pittii isolates are pathogenic and highly virulent to these organisms. Multilocus sequence typing analysis revealed that all the isolates belong to sequence type (ST) 839, which may be the dominant clone causing fish disease and exhibits a close phylogenetic relationship with ST396 from human clinical samples in Korea or Taiwan China. This is the first report demonstrating that A. pittii is an emerging causal agent of mass mortality in loach and poses significant risks to fish culturing besides causing human clinical infection worldwide.

A hypervirulent A. hydrophila (vAh) pathotype has been identified as the etiologic agent responsible for disease outbreaks in farmed carp species and channel catfish (Ictalurus punctatus) in China and the Southeastern United States, respectively. The possible route of infection has previously been unknown; however, virulence is believed to be multifactorial, involving the production/secretion of several virulence factors, including a high molecular weight group 4 capsular polysaccharide. Here we present chemical structural evidence of a novel capsule- and LPS-associated O-antigen found present in vAh isolated during these disease outbreaks. In this study, the chemical structure of the vAh O-antigen was determined by chemical analysis, Smith degradation, mass spectrometry, and 2D proton and carbon nuclear magnetic resonance (NMR) spectroscopy and found to be unique among described bacterial O-antigens. The O-antigen consists of hexasaccharide repeating units featuring a 4)-α- l -Fuc p -(1–3)-β- d -Glc p NAc-(1–4)-α- l -Fuc p -(1–4)-β- d -Glc p -(1- backbone, substituted with single residue side chains of α- d -Glc p and α- d -Qui p 3NAc linked to O-3 of the two fucose residues. The polysaccharide is partially O-acetylated on O-6 of the 4-substituted β-Glc p residue. Image 1 • The fish pathogen, A. hydrophila ST251, produces a high-molecular weight LPS. • This bacterium features a group 4 capsule comparable to the LPS O-antigen. • The repeat unit consists of a hexasaccharide of unique structure. [ABSTRACT FROM AUTHOR]