Effects of mosquito control using the microbial agent Bacillus thuringiensis israelensis (Bti) on aquatic and terrestrial ecosystems: a systematic review


The bacterium Bacillus thuringiensis serovar israelensis (Bti) is commercially produced in various formulations for use as a larvicide worldwide, targeting especially the aquatic larval stage of mosquitoes. However, there is a concern that repeated Bti treatments may have both direct and indirect impacts on non-target organisms (NTOs) and the ecosystems they inhabit. This review evaluates the evidence for such impacts.


Literature was searched using six bibliographic databases, two search engines, and on specialist web sites. Eligibility screening was performed in two steps on (1) title/abstract, with consistency among reviewers assessed by double-screening 557 articles and (2) full text. Articles included after full text screening were critically appraised independently by two reviewers. Disagreements were reconciled through discussions. Key parameters of included studies are presented in narrative synthesis tables, including risk of bias assessments. Meta-analyses comparing treated with untreated ecosystems and using either the raw mean difference or log response ratio as effect size were performed.

Review findings

Ninety-five articles covering 282 case studies were included in the review. From these, we identified 119 different response variables, which were divided into nine outcome categories. Most studies investigated NTO abundance or life history (reproduction related outcomes), but diversity and community composition are also well represented as outcome categories. The studies are highly variable in methodology, rigor, and spatio-temporal scale, spanning 1 day to 21 years and from < 1m2 to > 10,000 m2. Our metanalyses revealed a consistent negative effect of Bti treatment on abundances of Chironomidae and Crustacea, and also on chironomid emergence, although from a more restricted set of studies and regions. For most remaining response variables, we judged meta-analysis unfeasible, due to low study numbers or insufficient reporting of methods and results.


There is now a significant body of studies documenting effects of mosquito control using Bti on NTOs or other ecosystem properties, especially associated with negative effects on Chironomidae, as apparent from our meta-analyses. Accordingly, we suggest the potential for negative NTO or other ecosystem effects of Bti treatment should not be discounted a priori. Once a decision to proceed with Bti treatment has been taken, priority should be given to a well-designed program of ongoing monitoring and assessment. The paucity of rigorous studies conducted with low bias risk for most response variables undermines our capacity for evaluating how common many of the effects documented might be. Future research would benefit from a rigorous and well-replicated approach to studying Bti impacts in semi-field mesocosms or in the field, combined with a greater rigor in reporting key methodological details. A greater focus is needed on understanding the environmental factors which regulate the wider effects of mosquito control using Bti on NTOs and ecosystems, to enhance our capacity for predicting where and when Bti is most likely to have additional, negative and indirect ecological impacts.


The bacterium Bacillus thuringiensis serovar israelensis (Bti) is used in many countries as a biological larvicide to control dipteran insects of the suborder Nematocera, especially mosquitoes and black flies. Bti is generally accepted to be target-specific and efficient, with low potential for development of resistance among the target species. However, even though Bti may have minimal direct effects on non-target organisms, it might potentially be associated with knock-on effects on food webs and other ecosystem properties, including biodiversity and ecosystem functioning. Evidence from previous research is mixed, with some finding no evidence for indirect effects on biodiversity and ecosystem-level properties, but others indicating that such effects are possible. The fact that many studies have been conducted by organisations coordinating the control programs, and that many of those studies have been published outside peer reviewed scientific journals, highlights the challenges for decision makers and others to assess the results of the existing studies. In this protocol we outline how we aim to systematically and transparently synthesise all available evidence in a forthcoming systematic review.


We will use six bibliographic databases/platforms and the online search engines Google and Google Scholar in searches for literature. Searches will also be made on specialist websites. We will screen the search results for eligibility in stage one based on title and abstract, and in stage two based on the full text of the material. At stage one, after testing and clarifying the eligibility criteria, two reviewers will split and single screen the search results. At stage two the articles will be screened independently by two reviewers. We have developed a preliminary critical appraisal tool that will be used as basis for assessing study validity. Each study will be critically appraised independently by two reviewers. Disagreements will be reconciled through discussions seeking to reach consensus. It is unclear whether a quantitative synthesis based on meta-analysis will be feasible. A narrative synthesis will include descriptive statistics outlining the evidence base in terms of bibliographic information and study metadata. A narrative synthesis table in the form of an Excel spreadsheet will be provided.


Biocontrol, Biodiversity, Biopesticide, Culcidae, Food web, Indirect impacts, Insecticide, Larvicide, Simuliidae, Species Interactions