Background Adult anopheline mosquitoes transmit Plasmodium parasites that cause malaria. Some fish species eat mosquito larvae and pupae. In disease control policy documents, the World Health Organization includes biological control of malaria vectors by stocking ponds, rivers, and water collections near where people live with larvivorous fish to reduce Plasmodium parasite transmission. The Global Fund finances larvivorous fish programmes in some countries, and, with increasing efforts in eradication of malaria, policy makers may return to this option. We therefore assessed the evidence base for larvivorous fish programmes in malaria control. Objectives Our main objective was to evaluate whether introducing larvivorous fish to anopheline breeding sites impacts Plasmodium parasite transmission. Our secondary objective was to summarize studies evaluating whether introducing larvivorous fish influences the density and presence of Anopheles larvae and pupae in water sources, to understand whether fish can possibly have an effect. Search methods We attempted to identify all relevant studies regardless of language or publication status (published, unpublished, in press, or ongoing). We searched the following databases: the Cochrane Infectious Diseases Group Specialized Register; the Cochrane Central Register of Controlled Trials (CENTRAL), published in The Cochrane Library; MEDLINE; EMBASE; CABS Abstracts; LILACS; and the metaRegister of Controlled Trials (mRCT) until 18 June 2013. We checked the reference lists of all studies identified by the above methods. We also examined references listed in review articles and previously compiled bibliographies to look for eligible studies. Selection criteria Randomized controlled trials and non-randomized controlled trials, including controlled before-and-after studies, controlled time series and controlled interrupted time series studies from malaria-endemic regions that introduced fish as a larvicide and reported on malaria in the community or the density of the adult anopheline population. In the absence of direct evidence of an effect on transmission, we carried out a secondary analysis on studies that evaluated the effect of introducing larvivorous fish on the density or presence of immature anopheline mosquitoes (larvae and pupae forms) in community water sources to determine whether this intervention has any potential in further research on control of malaria vectors. Data collection and analysis Three review authors screened abstracts and examined potentially relevant studies by using an eligibility form. Two review authors independently extracted data and assessed risk of bias of included studies. If relevant data were unclear or were not reported, we wrote to the trial authors for clarification. We presented data in tables, and we summarized studies that evaluated the effects of fish introduction on anopheline immature density or presence, or both. We used GRADE to summarize evidence quality. We also examined whether the authors of included studies reported on any possible adverse impact of larvivorous fish introduction on non-target native species. Main results We found no reliable studies that reported the effects of introducing larvivorous fish on malaria infection in nearby communities, on entomological inoculation rate, or on adult Anopheles density. For the secondary analysis, we examined the effects of introducing larvivorous fish on the density and presence of anopheline larvae and pupae in community water sources. We included 12 small studies, with follow-up from 22 days to five years. Studies were conducted in a variety of settings, including localized water bodies (such as wells, domestic water containers, fishponds, and pools; six studies), riverbed pools below dams (two studies), rice field plots (three studies), and water canals (two studies). All studies were at high risk of bias. The research was insufficient to determine whether larvivorous fish reduce the density of Anopheles larvae and pupae (nine studies, unpooled data, very low quality evidence). Some studies with high stocking levels of fish seemed to arrest the increase in immature anopheline populations, or to reduce the number of immature anopheline mosquitoes, compared with controls. However, this finding was not consistent, and in studies that showed a decrease in immature anopheline populations, the effect was not consistently sustained. Larvivorous fish may reduce the number of water sources withAnopheles larvae and pupae (five studies, unpooled data, low quality evidence). None of the included studies reported effects of larvivorous fish on local native fish populations or other species. Authors' conclusions Reliable research is insufficient to show whether introducing larvivorous fish reduces malaria transmission or the density of adult anopheline mosquito populations. In research examining the effects on immature anopheline stages of introducing fish to potential malaria vector breeding sites (localized water bodies such as wells and domestic water sources, rice field plots, and water canals) weak evidence suggests an effect on the density or presence of immature anopheline mosquitoes with high stocking levels of fish, but this finding is by no means consistent. We do not know whether this translates into health benefits, either with fish alone or with fish combined with other vector control measures. Our interpretation of the current evidence is that countries should not invest in fish stocking as a larval control measure in any malaria transmission areas outside the context of carefully controlled field studies or quasi-experimental designs. Research could also usefully examine the effects on native fish and other non-target species.