First isolation and characterization of the pathogenic Aeromonas veronii bv. veronii associated with ulcerative syndrome in the indigenous Pelophylax ridibundus of Al-Ahsaa, Saudi Arabia

Abstract

Virulent microbial pathogen infections are the main cause for amphibian decline worldwide. In the present study, a bacterial strain named RDL-2, which was isolated from the skin of infected Pelophylax ridibundus larvae, was cultured and then phenotypically and biochemically characterized using scanning electron microscopic observations and the API20E strip and Biolog Gen III MicroPlate system. The 16S rRNA gene sequence of this strain was also obtained and used in strain identification and phylogenetic analysis. Healthy P. ridibundus larvae were challenged with RDL-2 and monitored to determine clinical signs consistent with the disease. Strain RDL-2 was identified as Aeromonas veronii based on its phenotypic and biochemical characteristics and on 16S rRNA gene sequence and phylogenetic analysis: RDL-2 was gram-negative, rod-shaped, and reacted positively for 69 (73%) of the 94 traits analysed; its 16S rRNA gene sequence displayed the highest homology to that of A. veronii bv. veronii (99.9%). Koch's postulates were fulfilled confirming that A. veronii is the causal agent of ulcerative syndrome. Thus, this study is the first to report A. veronii as a marsh frog pathogen in the Al-Ahsaa region. The aetiology of A. veronii as a potential poikilothermic pathogen shown here will expedite the development of diagnostic tests and methods for eradicating ulcerative syndrome.

Keywords: 16S rRNA; Aeromonas veronii; Al-Ahsaa; Larval mortality; Pelophylax ridibundus; Ulcerative Syndrome.