In the study a new aqueous formulation with the new mosquitocidal isolate of Bacillus cereus VCRC 641 was developed, and the residual activity was assessed from simulated field trials conducted in the natural environment. From laboratory bioassays, it was observed that the efficacy of aqueous formulation of Bacillus cereus was on par with lyophilized cell mass, as such. The LC50 values of formulated B. cereus against Culex quinquefasciatus, Anopheles stephensi and Aedes aegypti were 0.002, 0.009 and 0.008 mg L-1 respectively. Correspondingly, the LC50 values of lyophilized cell mass of B. cereus against these three mosquito species were 0.0019, 0.005 and 0.004 mg L-1 respectively. Finally, simulated field trial was carried out using formulated B. cereus and the residual efficacy against all three mosquito larval species revealed 100 per cent larval mortality up to seven days and 69 to 78 per cent mortality were up to 15 days. Thereafter, the mortality was declining gradually up to 21 days. It is concluded that the formulation of B. cereus may be used for mosquito control program.
In this study, search for new mosquitocidal bacterium was attempted from the gut content of the fresh water fish Clarias batrachus (walking cat fish) which resulted in the isolation of a highly potential mosquitocidal bacterium. The isolate was identified as Bacillus cereus VCRC 641 by Bacillus species specific molecular markers ilvD, pur and pycA gene sequence analysis. Laboratory mosquito larval toxicity assay (bioassay) against mosquito larvae revealed that this new isolate was highly potent against Culex quinquefasciatus followed by Aedes aegypti and Anopheles stephensi. The lethal concentration values at LC50 for these three mosquito species were 0.001, 0.004 and 0.007mg/ L respectively. Therefore, in the present study, this is the first report that a highly potential mosquitocidal bacteria of B. cereus VCRC 641 was isolated from the gut content of Clarias batrachus and suggested that it can be used for mosquito vector control program.
Mosquito vector-borne disease which accounts for the transmission of many deadly diseases like dengue, zika, chikungunya, Rift Valley fever, malaria, West Nile fever, heartworm, lumpy skin disease is one of the major causes of mortality, morbidity, and economic loss to human and animal communities. Though there have been several interventions like insecticide-treated nets, spatial repellents, indoor pesticide sprays to control adult mosquitoes, it only remains effective for short time, possibly leading to faster replacement of the adults by immature only achieving lesser mosquito population reduction. Extensive use of synthetic insecticides leads to resistance, affects non-target organisms and cause ecological imbalance. While recognizing the limitations of the synthetic and adulticide-based mosquito control methods, alternatively, microbial larvicidal agents are more feasible, target-specific, environment-friendly, low application rates, and moderate costs when compared with other vector control measures. This review highlights the microbial larvicidal agents as a most promising tool in vector control management to prevent disease transmission and discusses the major mosquito-borne diseases affecting humans and other animals.
Bacillus cereus VCRC 641 was found to be an alternate larvicidal bacterial agent which was newly isolated from fresh water fish, Clarias batrachus (walking cat fish). The protein responsible for mosquito toxic effect was separated and purified by Sephacryl S-200 column chromatography and NATIVE PAGE. It was observed that a unique protein with molecular weight of 70kDa was the responsible factor for mosquito killing effect. Further analysis of the protein by liquid chromatography-mass spectrometry (LC-MS) it was identified as “Uncharacterized Protein”. The length of the uncharacterized protein was around 158bp. The uncharacterized protein was expected to have a molecular mass of 70 kDa, 51 amino acids, confidence score of 78.45 and mass of 206772.57. Therefore, in the present study we report for the first time that uncharacterized protein (70kDa) was the responsible factor for the mosquito killing effect.
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