A silkworm infection model to investigate Vibrio vulnificus virulence genes

Yamamoto,Mai; Kashimoto,Takashige; Yoshimura,Yukihiro; Tachibana,Nao; Kuroda,Shiho; Miki,Yoshiko; Kitabayashi,Sou; Tong,Ping; Xiao,Jianbo; Tanaka,Koichi; Hamamoto,Hiroshi; Sekimizu,Kazuhisa; Yamamoto,Koichiro

doi:10.3892/mmr.2016.5782

A silkworm infection model to investigate Vibrio vulnificus virulence genes

Authors:
- Mai Yamamoto
- Takashige Kashimoto
- Yukihiro Yoshimura
- Nao Tachibana
- Shiho Kuroda
- Yoshiko Miki
- Sou Kitabayashi
- Ping Tong
- Jianbo Xiao
- Koichi Tanaka
- Hiroshi Hamamoto
- Kazuhisa Sekimizu
- Koichiro Yamamoto
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Affiliations: Department of Nutritional Science, Okayama Prefectural University, Soja, Okayama 719‑1197, Japan, School of Veterinary Medicine, Kitasato University, Towada, Aomori 034‑8628, Japan, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Bunkyo‑ku, Tokyo 113‑0033, Japan
Published online on: September 26, 2016 https://doi.org/10.3892/mmr.2016.5782
Pages: 4243-4247

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Abstract

The halophilic marine bacterium, Vibrio vulnificus, occasionally causes fatal septicemia in immunocompromised patients. Mice are commonly used as experimental animals to investigate the virulence of V. vulnificus, however, a large number of mice are generally required for bioassays. The present study examined whether the invertebrate species, silkworms, can be used instead of mice to investigate V. vulnificus virulence. When the silkworms were inoculated with 1.2x107 colony forming units of V. vulnificus OPU1‑Rf, a virulent strain of V. vulnificus, all injected silkworms died within 48 h, however, those injected with culture filtrate or diluent did not. This silkworm infection model was then used to isolate attenuated V. vulnificus mutants from 1,016 transposon‑inserted mutants. Consequently, a harmless mutant, SW998, was isolated. In this strain, the transposon was inserted into the rtxA gene, which is a known V. vulnificus virulence gene. In conclusion, the present study demonstrated that silkworms are useful animals for investigating the virulence of V. vulnificus.

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