Expression of Mosquito Active Toxin Genes by a Colombian Native Strain of the Gram-negative Bacterium Asticcacaulis excentricus

Mem Inst Oswaldo Cruz 2001, Volume 96, Number 2, pp. 257-263 Expression of Mosquito Active Toxin Genes by a Colombian Native Strain of the Gram-negative Bacterium Asticcacaulis excentricus Magally Romero, Flor M Gil, Sergio Orduz+ Code Number: oc01038 ABSTRACT: Mosquito control with biological insecticides, such as Bacillus sp. toxins, has been used widely in many countries. However, rapid sedimentation away from the mosquito larvae feeding zone causes a low residual effect. In order to overcome this problem, it has been proposed to clone the Bacillus toxin genes in aquatic bacteria which are able to live in the upper part of the water column. Two strains of Asticcacaulis excentricus were chosen to introduce the B. sphaericus binary toxin gene and B. thuringiensis subsp. medellin cry11Bb gene cloned in suitable vectors. In feeding experiments with these aquatic bacteria, it was shown that Culex quinquefasciatus, Aedes aegypti, and Anopheles albimanus larvae were able to survive on a diet based on this wild bacterium. A. excentricus recombinant strains were able to express both genes, but the recombinant strain expressing the B. sphaericus binary toxin was toxic to mosquito larvae. Crude protease A. excentricus extracts did not degrade the Cry11Bb toxin. The flotability studies indicated that the recombinant A. excentricus strains remained in the upper part of the water column longer than the wild type Bacillus strains. KEYWORDS: Asticcacaulis excentricus, Bacillus thuringiensis, recombinant bacteria, mosquito larvae Copyright 2001 Fundacao Oswaldo Cruz Fiocru