Identification, expression and bioactivity of Paramisgurnus dabryanus β-defensin that might be involved in immune defense against bacterial infection

•The first non-scaled freshwater fish β-defensin, pdBD, was identified in loach.•pdBD shared the conserved genomic structure with the reported fish with scales.•pdBD mRNA was induced in different tissues of Aeromonas hydrophila infected loach.•pdBD showed antibacterial activity against A. hydrophila and Bacillus subtilis.

β-defensins are a large family of multi-disulfide-bonded peptides with broad-spectrum antimicrobial activities that contribute to innate host defense in many organisms, but little information is available about β-defensins produced by freshwater fish lacking scales. We therefore cloned and identified a β-defensin gene from Chinese loach (Paramisgurnus dabryanus) by designing degenerate primers and using thermal asymmetric interlaced PCR. This gene is the first defensin gene ever identified in a non-scaled freshwater fish. Annotation of the protein domain architecture showed that the putative Chinese loach β-defensin contains the signature motif of six conserved cysteines within the mature peptide, an aspect similar to β-defensins of other marine fish. We also used quantitative real-time PCR to investigate the expression pattern of the Chinese loach β-defensin gene, mRNA of which could be observed in various tissues. After challenge with the pathogenic bacterium Aeromonas hydrophila, β-defensin expression was induced in the eye, gill, skin, and spleen of the adult loach. The bioactivity of the recombinant P. dabryanus β-defensin was examined against pathogenic bacteria, and the results suggest that this class 2 β-defensin has potential applications for treatment of bacterial infections.