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Journal of Biochemistry Advance Access originally published online on August 4, 2006
Journal of Biochemistry 2006 140(3):439-444; doi:10.1093/jb/mvj171
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© 2006 The Japanese Biochemical Society.

ARTICLE

Use of Silkworm Larvae to Study Pathogenic Bacterial Toxins

Muktadir S. Hossain*, Hiroshi Hamamoto, Yasuhiko Matsumoto, Iony M. Razanajatovo, Jorge Larranaga, Chikara Kaito, Hiroshi Kasuga and Kazuhisa Sekimizu{dagger}

Laboratory of Microbiology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 3-1, 7-chome, Hongo, Bunkyo-ku, Tokyo 113-0033

{dagger} To whom correspondence should be addressed. Tel: +81 3 5841 4820, Fax: +81 3 5684 2973, E-mail: sekimizu{at}mol.f.u-tokyo.ac.jp

Injection of stationary phase culture-supernatants of Staphylococcus aureus and Pseudomonas aeruginosa into the hemolymph of silkworm larvae caused their death, whereas a culture-supernatant of a non-pathogenic strain of Escherichia coli did not. A culture-supernatant of a mutant of agr, a global virulence regulator of S. aureus that is required for exotoxin production, was much less toxic to silkworm larvae. A culture-supernatant of a disruption mutant of the S. aureus beta-toxin gene did not kill larvae, whereas one of a deletion mutant of alpha-toxin, gamma-toxin, or aureolysin killed larvae, indicating that the beta-toxin gene is required for staphylococcal supernatant–mediated killing of silkworm larvae. The 50% lethal doses (LD50) of staphylococcal alpha-toxin and beta-toxin, Pseudomonas exotoxin A and diphtheria toxin were 12 µg/g, 9 µg/g, 0.14 µg/g and 1.1 µg/g, respectively. As the purified toxins killed the larvae, silkworm larvae could be used as a model to study the actions of pathogenic bacterial toxins in animal bodies.

* Present address: Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, MD20892, USA.


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