Bacillus thuringiensis (B.t.) δ-endotoxins provide an alternative to chemical insecticides for controlling many species of pest insects. Recent biotechnological developments offer the promise of even greater use of B.t. toxins in genetically transformed pest-resistant crops. However, the discovery that insects can adapt to these toxins raises concerns about the long-term usefulness of B.t. toxins. Several methods for managing the development of resistance to B.t. toxins have been suggested, but none of these approaches offer clear advantages in all situations.

http://science.sciencemag.org/content/sci/258/5087/1451.full.pdf

Managing Insect Resistance to Bacillus thuringiensis Toxins

Since the discovery of Bacillus thuringiensis (Berliner) serovariety israelensis de Barjac (Bti) and efficacious isolates of Bacillus sphaericus Neide, formulations of these bacteria have become the predominant non-chemical means employed for control of mosquito larvae at several locations in the United States and other countries. An overview of developments in the past 20 years is presented in this chapter regarding the toxins of Bti and B. sphaericus, their modes of action, efficacy and factors that affect larvicidal activity, development of resistance, safety, and their roles in integrated mosquito control. The efficacy of Bti formulations has been demonstrated in a variety of habitats against a multitude of species of mosquitoes. B. sphaericus formulations have been utilized predominantly in organically enriched habitats against Culex species, but they are also active in a variety of habitats having low organic enrichment, against numerous species, and across several genera. Stegomyia spp. are not susceptible to practical doses of B. sphaericus formulations. B. sphaericus has been shown to persist longer than Bti in polluted habitats and, under certain circumstances, can recycle in larval cadavers. A disadvantage of B. sphaericus has been the development of resistance in certain populations of Cx. quinquefasciatus Say and Cx. pipiens Linnaeus. Biotic and abiotic factors that influence the larvicidal activity of Bti and B. sphaericus include species of mosquito and their respective feeding strategies, rate of ingestion, age and density of larvae, habitat factors (temperature, solar radiation, depth of water, turbidity, tannin and organic content, presence of vegetation, etc.), formulation factors (type of formulation, toxin content, how effectively the material reaches the target, and settling rate), storage conditions, production factors, means of application and frequency of treatments. Due to their efficacy and relative specificity, both Bti and B. sphaericus can be ideal control agents in integrated programs especially where other biological control agents, environmental management, personal protection and the judicious use of insecticides are combined.

https://bioone.org/journals/journal-of-the-american-mosquito-control-association/volume-23/issue-sp2/8756-971X(2007)23[133:BTSIAB]2.0.CO;2/BACILLUS-THURINGIENSIS-SEROVARIETY-ISRAELENSIS-AND-BACILLUS-SPHAERICUS-FOR-MOSQUITO-CONTROL/10.2987/8756-971X(2007)23[133:BTSIAB]2.0.CO;2.pdf

Bacillus thuringiensis serovariety israelensis and bacillus sphaericus for mosquito control

https://mmbr.asm.org/content/mmbr/57/4/838.full.pdf

Mosquitocidal toxins of bacilli and their genetic manipulation for effective biological control of mosquitoes.

The impending widespread use of transgenic crop plants encoding a single insecticidal toxin protein of Bacillus thuringiensis has focused attention on the perceived risk of rapid selection of resistance in target insects. We have used Bacillus thuringiensis subsp. israelensis toxins as a model system and determined the speed and magnitude of evolution of resistance in colonies of the mosquito Culex quinquefasciatus during selection for 28 consecutive generations with single or multiple toxins. The parental strain was synthesized by combining approximately 500 larvae from each of 19 field collections obtained from the states of California, Oregon, Louisiana, and Tennessee. At least 10,000 larvae were selected in each generation of each line at an average mortality level of 84%. The susceptibilities of the parental and selected lines were compared in parallel tests in every third generation by using fresh suspensions of toxin powders. The normal toxin complement of B. thuringiensis subsp. israelensis consists of four toxins, CryIVA, CryIVB, CryIVD, and CytA. Resistance became evident first in the line that was selected with a single toxin (CryIVD), attaining the highest level (resistance ratio [RR], >913 at 95% lethal concentration) by generation F(inf28) when the study was completed. Resistance evolved more slowly and to a lower level (RR, >122 by F(inf25)) in the line selected with two toxins (CryIVA+CryIVB) and lower still (RR, 91 by F(inf28)) in the line selected with three toxins (CryIVA+CryIVB+ CryIVD). Resistance was remarkably low (RR, 3.2) in the line selected with all four toxins. The results reveal the importance of the full complement of toxins found in natural populations of B. thuringiensis subsp. israelensis as an effective approach to resistance management.

Influence of Exposure to Single versus Multiple Toxins of Bacillus thuringiensis subsp. israelensis on Development of Resistance in the Mosquito Culex quinquefasciatus (Diptera: Culicidae).

Cry proteins produced by Bacillus thuringiensis are selective biodegradable insecticides used increasingly in bacterial insecticides and transgenic plants as alternatives to synthetic chemical insecticides. However, the potential for development of resistance and cross-resistance in target insect populations to Cry proteins used alone or in combination threatens the more widespread use of this novel pest control technology. Here we show that high levels of resistance to CryIV proteins in larvae of the mosquito, Culex quinquefasciatus, can be suppressed or reduced markedly by combining these proteins with sublethal quantities of CytA, a cytolytic endotoxin of B. thuringiensis. Resistance at the LC95 level of 127-fold for a combination of three CryIV toxins (CryIVA, B, and D), resulting from 60 generations of continuous selection, was completely suppressed by combining sporulated powders of CytA in a 1:3 ratio with sporulated powders of a CryIVA, CryIVB, and CryIVD strain. Combining the CytA strain with a CryIVA and CryIVB strain also completely suppressed mosquito resistance of 217-fold to the latter toxins at the LC95 level, whereas combination of CytA with CryIVD reduced resistance in a CryIVD-selected mosquito strain from greater than 1,000-fold to less than 8-fold. The CytA/CryIV model provides a potential molecular genetic strategy for engineering resistance management for Cry proteins directly into bacterial insecticides and transgenic plants.

https://www.pnas.org/content/pnas/94/20/10536.full.pdf

Ira F. Goldman

Papers

Selection for resistance to Bacillus thuringiensis subspecies israelensis in field and laboratory populations of the mosquito Aedes aegypti.