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. 2023 Sep;16(9):1974-1984.
doi: 10.14202/vetworld.2023.1974-1984. Epub 2023 Sep 23.

Isolation, genetic characterization, and virulence profiling of different Aeromonas species recovered from moribund hybrid catfish (Clarias spp.)

Dini Siswani Mulia  1 , Rarastoeti Pratiwi  2 , Widya Asmara  3 , Mohamad Azzam-Sayuti  4 , Ina Salwany Md Yasin  4 , Alim Isnansetyo  5
Affiliations

Isolation, genetic characterization, and virulence profiling of different Aeromonas species recovered from moribund hybrid catfish (Clarias spp.)

Dini Siswani Mulia et al. Vet World. 2023 Sep.

Abstract

Background and aim: The high diversity of Aeromonas spp. results in various pathogenicity levels. This group of bacteria causes a serious disease named motile Aeromonas septicemia (MAS) in catfish (Clarias spp.). This study aimed to characterize the species and virulence gene diversity of Aeromonas spp. isolated from diseased catfish.

Materials and methods: Nine Aeromonas spp. were isolated from infected catfish cultivated in Java, Indonesia, and they were identified at the phenotypic and molecular levels (16S rDNA). The virulence genes assessed included aer/haem, alt, ast, flaA, lafA, and fstA.

Results: Phylogenetic analysis identified nine isolates of Aeromonas spp.: Aeromonas hydrophila (11.11%), Aeromonas caviae (11.11%), Aeromonas veronii bv. veronii (44.44%), and Aeromonas dhakensis (33.33%). Virulence genes, such as aer/haem, alt, ast, flaA, lafA, and fstA, were detected in all isolates at frequencies of approximately 100%, 66.67%, 88.89%, 100%, 55.56%, and 66.67%, respectively. This study is the first report on A. dhakensis recovered from an Indonesian catfish culture. Furthermore, our study revealed the presence of A. veronii bv veronii, a biovar that has not been reported before in Indonesia.

Conclusion: This finding confirms that MAS was caused by multiple species of Aeromonas, notably A. dhakensis and A. veronii bv veronii, within Indonesian fish culture. The presence of these Aeromonas species with multiple virulence genes poses a significant threat to the freshwater aquaculture industry.

Keywords: 16S rDNA; aeromoniasis; motile Aeromonas septicemia; pathogenicity; phenotype; phylogenetic.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure-1
Figure-1
Clinical signs of moribund hybrid catfish diseased. a: Depigmentation (D) of the skin, necrosis (N), b: Hemorrhagic (He), c: Hyperemia (Hi), d: Ulcer (U), e: Abdominal dropsy (AD), abdominal ascites (AA), f: Abscess (S), pale (P) in kidney, abdominal ascites (AA).
Figure-2
Figure-2
Results of amplification of bacterial DNA isolated with 16S rDNA primers. Arrow: 1500 bp, Marker: 1 kb.
Figure-3
Figure-3
Phylogenetic tree constructed from the 16S rDNA sequences of Aeromonas sp. and other Aeromonas species (class of Gammaproteobacteria) and detected virulence genes. Pseudomonas aeruginosa was used as an outgroup. The topology was obtained by maximum likelihood with bootstraps of 1000 replications. The scale bar signifies 0.02 substitutions per nucleotide position (Knuc). Colored circles indicate the presence of the virulence genes for each isolate, formula image aer/haem: aerolysin/hemolysin; formula image alt: heat-labile cytotonic enterotoxin; formula image ast: heat-stable cytotonic enterotoxin; formula image flaA: polar flagellum; formula image lafA: lateral flagellum; formula image fstA: ferric siderophore receptor.
See this image and copyright information in PMC

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