Documentation of high-level bacillus Sphaericus 2362 resistance in field populations of Culex quinquefasciatus breeding in polluted water in Thailand

Abstract

Bacillus sphaericus (Bsph) Neide has been recognized as an effective mosquito larvicide since its discovery 20 years ago. Various strains of this agent, such as 2362, 2297, 1593, and C3-41, have been developed, formulated, and field-evaluated against mosquito larvae in different countries. Their high efficacy in controlling mosquitoes breeding in various habitats, especially those in polluted water, has been documented. However, resistance to Bsph has been reported in Culex pipiens complex in both laboratory colonies and natural populations. During our field trials on Bsph water-dispersible granules (WDG) against natural populations of Cx. quinquefasciatus Say in a low-income community, Nonthaburi Province, Thailand, control failure occurred within 4 months after 5 treatments using VectoLex WDG at the dosages of 50-200 mg/m2. The suspected Bsph-resistant Cx. quinquefasciatus mosquitoes were collected and colonized in the laboratory, from which the late stage larvae of generations F14 through F17 were used in bioassays to elucidate the resistance profile. A high level of Bsph resistance was documented in this colony as compared with the susceptible mosquitoes from the same area in Thailand or from California, USA. The resistance ratios (RR) at LC50, depending on reference colonies, were 21,100-28,100-fold against Bsph WDG or greater than 125,000-200,000-fold against Bsph technical-grade material. These Bsph-resistant mosquitoes, however, were completely susceptible to Bacillus thuringiensis var. israelensis, (Bti) preparations, LC50 ranging from 0.017 ppm for technical material with 7000 ITU/mg to 0.052 ppm for water-dispersible granules with 3000 ITU/mg. The RR to mixtures of Bsph + Bti in this highly Bsph-resistant mosquitoes increased steadily upon the increase of Bsph ratios in the mixtures from 50, 75, 90, 95 to 99%. The resistance levels to the mixtures with various ratios of Bsph and Bti, however, were substantially lower than that in Bsph alone, suggesting addition of Bti to Bsph substantially enhanced the mosquitocidal activity (synergism) against these highly Bsph-resistant Cx. quinquefasciatus. Moderate tolerance to low levels of resistance to Bsph/Bti recombinant (RR 7.29-12.75 at LC50 and 5.15-13.40 at LC90) was also noted in this Bsph-resistant population.