Long-term persistence and recycling of Bacillus thuringiensis israelensis spores in wetlands sprayed for mosquito control.

Bacillus thuringiensis subsp. israelensis (Bti) is the main larvicide used to control mosquitoes worldwide. Although there is accumulating evidence of Bti having environmental effects on non-target fauna, relatively few field studies have documented the fate of Bti spores in the environment. Spore density was quantified over a 6-yr period (2012-2017) in Mediterranean marshes sprayed with Vectobac 12AS (32 ITU/ha) since 2006 to reduce the nuisance caused by Aedes caspius. Bti spores were naturally found in all habitat types. Spore density expressed as colony-forming units per gram of soil (CFU g) increased significantly at treated sites by a factor of 22 to 500 times relative to control sites, with mean values of 7730 CFU g in halophilous scrubs, 38,000 in reed beds, 49,000 in bulrush beds and 50 000 in rush beds. Spore density varied little in the first months after the spraying season (April-October), but increased sharply in spring, just before the annual launch of mosquito control. Considering that Bti is an insect pathogen that cannot proliferate without a suitable insect host, this unexpected recrudescence in spring could be related to the warming of water that triggers activity and development of benthic organisms such as chironomids, which may contribute to Bti proliferation by ingesting accumulated spores at the surface of sediments. While spore density tends to decrease over time, presumably during the summer period as a result of increased UV exposure, three to four years were necessary for spore density to return to normal levels after mosquito-control interruption. This study is important because it demonstrates that environmental effects of mosquito-control using Bti can far exceed the short period of Bti efficacy against lentic mosquitoes. Considering that Bti is a microbial agent, these long-term effects should be addressed at multiple levels of ecosystem organization from a one-health perspective.