Isolation of Vibrio cholerae and Vibrio vulnificus from Estuarine Waters, and Genotyping of V. vulnificus Isolates Using Loop-Mediated Isothermal Amplification

• Published in: Microorganisms (Basel) 2024-05, Vol.12 (5), p.877
• Main Authors: Miyoshi, Shin-Ichi, Kurata, Megumi, Hirose, Riho, Yoshikawa, Masaya, Liang, Yong, Yamagishi, Yosuke, Mizuno, Tamaki
• Format: Article
• Language: English
• Subjects: Annealing; Aquaculture; Aquaculture facilities; Bacteria; Bacteria, Pathogenic; Chemical oxygen demand; Cholera; Coastal waters; Coliforms; Diarrhea; Disseminated infection; Eels; Environmental aspects; Estuaries; Gene amplification; Genes; Genetic aspects; Genetic testing; genotype; Genotype & phenotype; Genotypes; Genotyping; Gram-negative bacteria; Identification and classification; Infectious diseases; LAMP; Physiological aspects; Polymerase chain reaction; Salinity; Sepsis; Septicemia; Shellfish; Strains (organisms); Toxins; Vibrio; Vibrio cholerae; Vibrio vulnificus; Vibriosis; Water analysis; Water sampling; Water temperature; Waterborne diseases
• ISSN: 2076-2607; 2076-2607
• DOI: 10.3390/microorganisms12050877

Bacteria in the genus are ubiquitous in estuarine and coastal waters. Some species (including and are known human pathogens causing ailments like cholera, diarrhea, or septicemia. Notably, can also cause a severe systemic infection (known as vibriosis) in eels raised in aquaculture facilities. Water samples were periodically collected from the estuary of the Asahi River, located in the southern part of Okayama City, Japan. These samples were directly plated onto CHROMagar Vibrio plates, and colonies displaying turquoise-blue coloration were selected. Thereafter, polymerase chain reaction was used to identify and . A total of 30 strains and 194 strains were isolated during the warm season when the water temperature (WT) was higher than 20 °C. Concurrently, an increase in coliforms was observed during this period. Notably, has two genotypes, designated as genotype 1 and genotype 2. Genotype 1 is pathogenic to humans, while genotype 2 is pathogenic to both humans and eels. The loop-mediated isothermal amplification method was developed to rapidly determine genotypes at a low cost. Of the 194 strains isolated, 80 (41.2%) were identified as genotype 1 strains. Among the 41 strains isolated when the WTs were higher than 28 °C, 25 strains (61.0%) belonged to genotype 1. In contrast, of the 32 strains isolated when the WTs were lower than 24 °C, 27 strains (84.4%) belonged to genotype 2. These results suggest that the distribution of the two genotypes was influenced by WT.