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Synergisitic effect of chitinases and Bacillus thuringiensis israelensis spore-toxin complex against Aedes aegypti larvae

Published online by Cambridge University Press:  03 January 2012

Montserrat Ramírez-Suero ,
Gerardo Valerio-Alfaro ,
Julio S. Bernal  and
Mario Ramírez-Lepe
Montserrat Ramírez-Suero
Affiliation:
Unidad de Investigación y Desarrollo en Alimentos-Instituto Tecnológico de Veracruz, M.A. de Quevedo 2779, Veracruz, Veracruz 91897, México
Gerardo Valerio-Alfaro
Affiliation:
Unidad de Investigación y Desarrollo en Alimentos-Instituto Tecnológico de Veracruz, M.A. de Quevedo 2779, Veracruz, Veracruz 91897, México
Julio S. Bernal
Affiliation:
Biological Control Laboratory, Department of Entomology, Texas A&M University, College Station, Texas 77843-2475, United States of America
Mario Ramírez-Lepe*
Affiliation:
Unidad de Investigación y Desarrollo en Alimentos-Instituto Tecnológico de Veracruz, M.A. de Quevedo 2779, Veracruz, Veracruz 91897, México
*
1Corresponding author (e-mail: mario.ramirez.lepe@gmail.com).
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Abstract

Six subspecies of Bacillus thuringiensis Berliner (Bt) were grown in minimal medium with chitin as the sole carbon source for 6 days to obtain Bt cell-free fermented broths, which were then evaluated for chitinolytic activity and tested against third-instar Aedes aegypti (L.) (Diptera: Culicidae) larvae. Bt pakistani showed the highest chitinolytic activity (approximately >2700 mU/mL), Bt kurstaki showed the lowest activity (approximately <2000 mU/mL), and Bt thompsoni, Bt aizawai, Bt israelensis, and Bt alesti showed intermediate activities (approximately 2100–2400 mU/mL). Bt aizawai and Bt thompsoni broths showed the highest toxicity (LC50) against third-instar A. aegypti larvae (approximately <290 mU/mL). Bt kurstaki broth showed the lowest toxicity (approximately 420 mU/mL), while Bt pakistani, Bt israelensis, and Bt alesti broths showed intermediate toxicities (approximately 360–460 mU/mL). A purified and biochemically characterized Bt aizawai chitinase and commercial chitinases (from Serratia marcescens Bizio and Streptomyces griseus Waksman and Henrici) were evaluated and compared for synergistic effects on Bt israelensis spore-toxin complex against third-instar A. aegypti larvae. The synergism factor value of Streptomyces griseus and Bt aizawai chitinases were >2 and approximately 1.4; synergism was not evident for the Serratia marcescens chitinase (synergism factor value approximately 0.9).

Résumé

Six sous-espèces de Bacillus thuringiensis Berliner (Bt) ont été cultivées en milieu minimal avec la chitine comme seule source de carbone pendant 6 jours afin d’obtenir un surnageant sans cellules Bt. L’activité chitinolytique et la toxicité sur le troisième stade larvaire d’Aedes aegypti (L) (Diptera : Culicidae) de ce surnageant ont été mesurés. Des six sous-espèces testées, Bt pakistani a montré l’activité chitinolytique la plus élevée (> approximativement 2700 mU/mL), alors que Bt kurstaki a montré l’activité chitinolytique la plus faible (< approximativement 2000 mU/mL), et Bt thompsoni, Bt aizawai, Bt israelensis et Bt alesti ont montré des activités intermédiaires (approximativement 2100–2400 mU/mL). Les surnageants de Bt aizawai et Bt thompsoni ont montré la toxicité plus élevée (LC50) pour le troisième stade larvaire d’A. aegypti (< approximativement 290 mU/mL), alors que Bt kurstaki possédait la toxicité la plus faible (approximativement 420 mU/mL) et que Bt pakistani, Bt israelensis, et Bt alesti avaient des toxicités intermédiaires (approximativement 360–460 mU/mL). Une chitinase purifiée et biochimiquement caractérisée, Bt aizawai et des chitinases commerciales (provenant de Serratia marcescens Bizio et Streptomyces griseus Waksman and Henrici) ont été évaluées et les effets synergiques mesurés sur le complexe de spores et toxines Bt israelensis contre le troisième stade larvaire d’A. aegypti. La valeur du facteur de synergie des chitinases de Streptomyces griseus et Bt aizawai étaient de > 2 et approximativement 1,4 respectivement, et aucune synergie n’a été mise en évidence pour la chitinase de Serratia marcescens (approximativement 0,9).

Type
Articles
Information
The Canadian Entomologist , Volume 143 , Issue 2 , April 2011 , pp. 157 - 164
Copyright
Copyright © Entomological Society of Canada 2011

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