Isolation And Characterization Of A Novel  Bacterium, Bacillus Thuringiensis Var. Israelensis Vcrc-B651  Exhibiting High Efficacy To Controlmosquito Vectors

P Hemaladkshmi; Sahadiya Mandodan; Bhagyashree Bora; S  Manikandan; V Abhisubesh; Jibi Lukose; K Aneha; Kakhuangailiu Gangmei; A Mathivanan,; K Vijayalakshmi; S Poopathi

doi:10.48165/

Authors

P Hemaladkshmi Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)
Sahadiya Mandodan Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)
Bhagyashree Bora Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)
S Manikandan Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)
V Abhisubesh Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)
Jibi Lukose Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)
K Aneha Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)
Kakhuangailiu Gangmei Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)
A Mathivanan, Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)
K Vijayalakshmi Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)
S Poopathi Unit of Microbiology and Immunology, Vector Control Research Centre, Indian Council of Medical Research, Department of Health Research, Ministry of Health and Family Welfare, Puducherry – 605 006 (India)

DOI:

https://doi.org/10.48165/

Keywords:

Bacillus thuringiensis serovar israelensis, bioassays, clay soil, growth dynamics, mosquito larvae, whole genome sequence

Abstract

The aim of this study was to isolate novel mosquitocidal bacteria from clayey soil. The soil samples were collected during the period 2021 to 2022 from an agricultural field of Tamil Nadu (India). The samples were processed for isolating the bacteria on nutrient yeast extract salt medium. The bacterial cell mass was harvested after 72 h of lyophilisation. Bioassay was carried out with the bacterium against larval species of Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi. Whole genome sequence (WGS) analysis (Illumina) was done to identify the strain. Growth dynamics, biochemical characteristics, and toxicity assays with non-target aquatic organisms were also done. Based onWGS analysis the new isolate was identified as Bacillus thuringiensis serovar israelensis. The strain was coded as VCRC-B651. The isolate exhibited effective toxicity against A. aegypti, A.stephensi, and C. quinquefasciatus with LC50 values of 0.005, 0.009 and 0.009 mg L-1, respectively, and LC90 values of 0.009, 0.02 and 0.018 mg L-1, respectively. The isolate exhibited higher mosquitocidal activity as compared to the WHO reference strain (Bti H14). No adverse effect on aquatic non-target organisms was observed. The new isolate was highly efficacious in controlling the mosquito larvae.

Downloads

Download data is not yet available.

References

Abbott, W.S. 1925. A method of computing the effectiveness of an insecticide. Journal of Economical Entomology, 18: 256-267.

Asigau, S. and Parker, P.G. 2018. The influence of ecological factors on mosquito abundanceand occurrence in Galápagos. Journal of Vector Ecology, 43: 125-137.

Benelli, G., Jeffries, C.L. and Walker, T. 2016. Biological control of mosquito vectors: Past, present, and future. Insects, 7: 52. [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5198200/]. Bradford, M.M. 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72: 248-254. Caminade, C., McIntyre, K.M. and Jones, A.E. 2019. Impact of recent and future climate change on vector-borne diseases. Annals of the New York Academy of Sciences, 1436: 157-173. da Silva W.J., Pilz-Júnior, H.L., Heermann, R. and da Silva, O.S. 2020. The great potential of entomopathogenic bacteria Xenorhabdus and Photorhabdus for mosquito control: A review. Parasites and Vectors, 13: 1-4.

Dahmana, H. and Mediannikov, O. 2020. Mosquito-borne diseases emergence/resurgence andhow to effectivelycontrolit biologically. Pathogens, 9: 310.[https://www.mdpi.com/20760817/9/4/310]. Doggett, N.A., Stubben, C.J., Chertkov, O., Bruce, D.C., Detter, J.C., Johnson, S.L. and Han, C.S. 2013. Complete genome sequence of Bacillus thuringiensis serovar israelensis strain HD 789. Genome Announcements, 1(6): e01023-13 [doi:10.1128/genomeA.01023-13]. Federici, B.A., Park, H.W. and Bideshi, D.K. 2010. Overview of the basic biology of Bacillus thuringiensis with emphasis on genetic engineering of bacterial larvicides for mosquito control. Open Toxicology Journal. 3: 83-100.

Filha, M.H.N.L.S., Berry, C. and Regis, L. 2014. Lysinibacillus sphaericus: Toxins and mode of action, applications for mosquito control and resistance management. Advances in Insect Physiology, 47: 89-176.

Folly, A.J., Dorey-Robinson, D., Hernández-Triana, L.M., Phipps, L.P. and Johnson, N. 2020. Emerging threats to animals in the United Kingdom by arthropod-borne diseases. Frontiersin Veterinary Science 7: 20 [https://www.ncbi.nlm.nih.gov/pmc/articles/ PMC7010938/].

P. Hemaladkshmi et al.

Fournet, F., Jourdain, F., Bonnet, E., Degroote, S. and Ridde, V. 2018. Effective surveillance systems for vector-borne diseases in urban settings and translation of the data into action: A scoping review. Infectious Diseases of Poverty, 7: 1-4.

Geetha, I., Prabakaran, G., Paily, K.P., Manonmani, A.M. and Balaraman, K. 2007. Characterisation of three mosquitocidal Bacillus strains isolated from mangrove forest. Biological Control, 42: 34-40.

Joshi, A. and Miller, C. 2021. Review of machine learning techniques for mosquito control inurban environments. Ecological Informatics, 61: 101241. [https://www.sciencedirect.com/science/ article/pii/S1574954121000327].

Joshi, V., Sharma, R.C., Singhi, M., Singh, H., Sharma, K., Sharma, Y. and Adha, S. 2005. Entomological studies on malaria in irrigated and non-irrigated areas of Thar desert, Rajasthan, India. Journal of Vector Borne Diseases, 42: 25. [http://www.mrcindia.org/journal/issues/ 421025.pdf].

Kumar, D.P., Arti Mathur, Priyansh Sharma, Esha Shrivastava and Megha. 2012. Soil bacteria and their possible role in mosquito control: A short review. World Journal of Environmental Biosciences, 2: 40-48.

Laemmli, U.K. 1970. Cleavage of structural proteins during assembly of head bacteriophage T4. Nature; 227: 680-685.

Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R. 1951.JProtein measurement withthe Folin phenol reagent. The Journal of Biological Chemistry, 193: 265. [https://pubmed.ncbi.nlm.nih.gov/14907713/].

Mani, C., Thirugnanasambantham, K., Sundarapandian, S. and Poopathi, S. 2015. Identification and characterization of a novel marine Bacillus cereus VCRC-B540 for mosquito control. BioControl, 60: 71-79.

Mani, C., Selvakumari, J., Han, Y., Jo, Y., Thirugnanasambantham, K., Sundarapandian, S. and Poopathi, S. 2018. Molecular characterization of mosquitocidal toxin surface layer protein (SLP) from Bacillus cereus VCRC B540. Applied Biochemistry and Biotechnology, 184: 1094-1105.

Manikandan, S., Aneha, K., Bora, B., Abhisubesh, V., Hemaladkshmi, P., Gangmei, K., Mandodan, S., Lukose, J., Mathivanan, A., Vijayalakshmi, K. and Poopathi, S. 2023. Identification and characterization of mosquitocidal toxins from Bacillus cereus VCRC- 641. Indian Journal of Entomology, 23: [https://indianentomology.org/index.php/ije/article/view/833].

Mardhiah, I., Norhazilan, M.N., Nordin, Y., Akrima, A.B., Muhammad, K. and Arman, A. 2015. Statistical investigation on anaerobic sulphate-reducing bacteria growth by turbidity method. International Journal of Biological Chemistry, 9: 178-187.

Niang, E.H.A., Bassene, H., Fenollar, F. and Mediannikov, O. 2018. Biological control of mosquito borne diseases: The potential of Wolbachia-based interventions in an IVM Framework. Journal of Tropical Medicine, 1470459: [https://www.hindawi.com/journals/jtm/2018/1470459/abs/].

Poopathi, S., De Britto, L.J., Praba, V.L., Mani, C. and Praveen, M. 2015. Synthesis of silver nanoparticles from Azadirachta indica - A most effective method for mosquito control. Environmental Science and Pollution Research International, 22: 2956-2963.

Poopathi, S., Mani, C., Thirugnanasambantham, K., Praba, V.L., Ahangar, N.A. and Balagangadharan, K. 2014. Identification and characterization of a novel marine Bacillus cereus for mosquito control. Parasitology Research, 113: 323-332.

Poopathi, S., Mani, C., Vignesh, V., Praba, V.L. and Thirugnanasambantham, K. 2013. Genotypic diversity of mosquitocidal bacteria (Bacillus sphaericus, B. thuringiensis, and B. cereus) newly isolated from natural sources. Applied Biochemistry and Biotechnology, 171: 2233-2246.

Poopathi, S., Thirugnanasambantham, K., Mani, C., Ragul, K. and Sundarapandian, S.M. 2014. Isolation of mosquitocidal bacteria (Bacillus thuringiensis, B. sphaericus and B. cereus) from excreta of arid birds. Indian Journal of Experimental Biology, 52: 739-747.

Bt isolate from clay soil for mosquito control 457

Rabha, M., Das, D., Konwar, T., Acharjee, S. and Sarmah, B.K. 2023. Whole genome sequencing of a novel Bacillus thuringiensis isolated from Assam soil. BMC Microbiology, 23: 91. [https://bmcmicrobiol.biomedcentral.com/articles/10.1186/s12866-023-02821-0].

Ramirez, J.L., Short, S.M., Bahia, A.C., Saraiva, R.G., Dong, Kang, S., Tripathi, A., Mlambo, G. and Dimopoulos, G. 2014. Chromobacterium CspP reduces malaria and dengue infection in vector mosquitoes and has entomopathogenic and in vitro anti-pathogen activities. PLoS Pathogens, 10: e1004398. [https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1004398].

Rao, D.R., Mani, T.R., Rajendran, R., Joseph, A.S., Gajanana, A, and Reuben, R. 1995. Development of a high level of resistance to Bacillus sphaericus in a field population of Culex quinquefasciatus from Kochi, International Journal of American Mosquito Control Association, 11: 1-5.

Schaeffer, A.B. and Fulton, M.D. 1933. A simplified method of staining endospores. Science, 77: 1990. [https://pubmed.ncbi.nlm.nih.gov/17741261/].

Senthil-Nathan, S. 2020. A Review of resistance mechanisms of synthetic insecticides and botanicals, phytochemicals, and essential oils as alternative larvicidal agents against mosquitoes. Frontier Physiology, 10: 1591. [https://www.frontiersin.org/articles/10.3389/fphys.2019.01591].

Silva-Filha, M.H.N.L., Romão, T.P., Rezende, T.M.T., Carvalho, K.D.S., Gouveia de Menezes, H.S., Alexandre do Nascimento, N., Soberón, M. and Bravo, A. 2021. Bacterialtoxins active against mosquitoes: Mode of action and resistance. Toxins (Basel), 13: 523. [https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8402332/].

Wilson, A.L., Courtenay, O., Kelly-Hope, L.A., Scott, T.W., Takken, W., Torr, S.J. and Lindsay, S.W. 2020. The importance of vector control for the control and elimination of vector-borne diseases. PLoS Neglected Tropical Disease. 14: 1-31.

Wipfli, M. and Merritt, R. 1994. Disturbance to a stream food web by a bacterial larvicide specific to black flies: feeding responses of predatory macroinvertebrates. Freshwater Biology, 32: 91-103. Wirth, M.S., Walton, W.E. and Federici, B.A. 2010. Evolution of resistance to the Bacillus sphaericus

Bin toxin is phenotypically masked by combination with the mosquitocidal proteins of Bacillus thuringiensissubspeciesisraelensis. Environmental Microbiology, 12: 1154-1160. World Health Organization, 2005. Report of WHOPES on 324 Guidelines for Laboratory and Field Testing of Mosquito Larvicides, 325, World Health Organization, Geneva, Switzerland [WHO/CDS/WHOPES/GCDPP1].