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Objective:To investigate lethal effect of culture filtrates ofStreptomyces citreofluorescens (S. citreofluorescens)againstAnopheles stephensi (An. stephensi), Culex quinquefasciatus (Cx. quinquefasciatus), andAedes aegypti (Ae. aegypti) larvae vectors for malaria, filarial and dengue. Methods:The culture filtrates obtained fromS. citreofluorescens2528 was grown inPotato DextroseBroth(PDB), filtrated and used for the bioassay after a growth of15 days.Results:The results demonstrated that theAn. stephensi shows mortalities withLC50,LC90 values of first instar 46.8 μL/mL,79.5 μL/mL, second instar79.0μL/mL,95.6μL/mL, third instar79.0 μL/mL,136.9 μL/mL, and fourth instar122.6 μL/mL,174.5 μL/mL.Whereas,TheCx. quinquefasciatus were found effective on first instar40.0 μL/mL,138.03 μL/mL, second instar80.0 μL/mL,181.97 μL/mL, third instar100.0 μL/mL,309.2 μL/mL, and fourth instar60.0 μL/mL,169.82 μL/mL.The Ae. aegypti were successfully achieved susceptible with higher concentrations in comparisons ofAn. stephensi andCx. quinquefasciatus larvae.These outcomes of the investigations have compared with theChitinase of Streptomyces griseus (S. griseus)C6137 that shows90%-95% mortality.Conclusions:These new findings significantly permitted that the culture filtrates ofS. citreofluorescens can be used as bacterial larvicides.This is an environmentally safe approach to control the vectors of malaria, dengue and filariasis of tropical areas.