Optimization of Bacillus thuringiensis Growth Conditions for Enhanced Larvicidal Activity against Anopheles mosquitoes larvae
DOI:
https://doi.org/10.47430/ujmr.25101.008Keywords:
Bacillus thuringiensis, Control, Malaria, Mosquito larvae, OptimizationAbstract
Study’s Excerpt:
- The study optimised physicochemical parameters for growth and larvicidal activity of Bacillus thuringiensis.
- The optimal conditions are (Temperature = 29 °C, pH = 7.5, salinity= 0 w/v, inoculum volume = 20% v/v).
- Upon optimisation, 85% larvicidal activity was achieved in an eco-friendly manner.
- The isolates are recommended for use in eco-friendly biocontrol of insects.
Full Abstract:
One of the conventional ways of tackling malaria is the application of insecticides to mosquito breeding places. The study’s potential contributions to knowledge include providing a comprehensive understanding of how to maximize the effectiveness of Bti extracts, which can be applied in various ecological contexts and potentially lead to more effective mosquito control strategies. This study bridges the gap in identifying the optimal growth conditions for Bacillus thuringiensis to enhance its larvicidal activity against Anopheles mosquitoes larvae. This study aimed at enhancing the potential of Bacillus thuringiensis in the control of mosquito larvae. Bacillus thuringensis was isolated from soil samples in Katsina metropolis. Optimization of the culture conditions for the growth of the bacterium was carried out using selected parameters (pH, temperature, salinity and inoculum volume ) based on single factor at a time analyses. Mosquito larvae were obtained from Kofar Marusa water outlet, Katsina and characterized based on morphological and larvicidal stages (instar). The larvae mortality test was carried out using the prepared inocula of B. thuringensis, and the mortality rate among the larvae was recorded at intervals. The results obtained indicate that the population range of Bacillus thuringiensis ranged from 2.01 × 103 ± 0.03 CFU/mL to 3.62 × 103 ± 0.02 CFU/mL. The highest and lowest percentage mortality obtained were 85% and 50% respectively. The initial culture conditions used involved a temperature of 29 0C, a pH of 7.5, a salinity of 2% and inoculum volume of 1% v/v. The optimum conditions achieved involved a temperature of 30 0C, pH value of 7.5, salinity of 0% w/v and inoculum volume of 20% v/v. This indicates that B. thuringiensis represents an effective alternative to chemical insecticides in the control of mosquito larvae. Considering the fact that Bt toxins are safe for non-target species and human health, and so far, no resistance among the target has been detected, coupled with their specificity and eco-friendly nature, this study recommends using integrated pest management campaigns involving the use of bioinsecticides to serve as a better alternative to chemical insecticides which are harmful to the ecosystem.
Downloads
References
Amin, M., Rakhisi, Z., & Ahmady, A. Z. (2015). Isolation and identification of Bacillus species from soil and evaluation of their antibacterial properties. Avicenna Journal of Clinical Microbiology and Infection, 2(1), 23233. https://doi.org/10.17795/ajcmi-23233
Andrzejczak, S., & Lonc, E. (2008). Selective isolation of Bacillus thuringiensis from soil by use of L-serine as minimal medium supplement. Polish Journal of Microbiology, 5(4), 333–335. http://www.pjmonline.org/wp-content/uploads/2015/12/vol5742008333
Bahrami, R., Quaranta, S., Perdomo, H. D., Bonizzoni, M., & Khorramnejad, A. (2024). Carry-over effects of Bacillus thuringiensis on tolerant Aedes albopictus mosquitoes. Parasites & Vectors, 17(1). https://doi.org/10.1186/s13071-024-06556-3
Bravo, A., Likitvivatanavong, S., Gill, S. S., & Soberon, M. (2011). Bacillus thuringiensis: A story of a successful bioinsecticide. Insect Biochemistry and Molecular Biology, 41(7), 423–431. https://doi.org/10.1016/j.ibmb.2011.02.006
Bishop, A., & Robinson, C. (2014). Bacillus thuringiensis HD-1 Cry-: Development of a safe, non-insecticidal simulant for Bacillus anthracis. Journal of Applied Microbiology, 117(3), 654–662. https://doi.org/10.1111/jam.12560
Da Costa, A., Bonner, M., Rao, S., Gilbert, L., Sasaki, M., Elsey, J., MacKelfresh, J., & Arbiser, J. (2020). Establishment of a temperature-sensitive model of oncogene-induced senescence in angiosarcoma cells. Cancers, 12(2), 395. https://doi.org/10.3390/cancers12020395
Duarte Neto, J. M. W., Wanderley, M. C. D. A., da Silva, T. A. F., et al. (2020). Bacillus thuringiensis endotoxin production: A systematic review of the past 10 years. World Journal of Microbiology and Biotechnology, 36(8), 128. https://doi.org/10.1007/s11274-020-02904-4
Ennouri, K., Ayed, R. V., & Hassen, B. H. (2015). Mathematical screening of nutritional parameters for enhancement of insecticidal proteins production by Bacillus sp. Journal of New Sciences, Agriculture and Biotechnology, 16(2), 552–558.
Glare, T. R., & O'Callaghan, M. (2000). Bacillus thuringiensis: Biology, ecology and safety. John Wiley & Sons.
Gomis-Cebolla, J., dos Santos, F. R., Wang, Y., Cabellero, J., Caballero, P., He, K., Jurat-Fuentes, J. L., & Ferre, J. (2020). Domain shuffling between Vip3A and Vip3Ca: Chimera stability and insecticidal activity against European, African and Asian pests. Toxins, 12(9), 599. https://doi.org/10.3390/toxins12020099
Gupta, S., & Dikshit, A. K. (2010). Biopesticides: An ecofriendly approach for pest control. Journal of Biopesticides, 3(1), 186–188.
Hamedo, H. (2016). Identification of Bacillus thuringiensis isolated from different sources by Biology GEN III system and scanning electron microscopy. International Journal of Environmental Science, 15(1), 51–57. https://cat.journals.ekb.eg/article_18336_122fdcac7601e9e877ea5599bb0593b0.pdf
Haruna, M. Y., Khan, A. H., & Umar, Z. D. (2014). Effect of acid rain on growth of papaya (Carica papaya) and castor (Ricinus communis) plants. *e-Journal of Science and Technology, 10*(1), 43–47.
Hernandez, E., Ramisse, F., Gros, P., & Cavallo, J. (2000). Super-infection by Bacillus thuringiensis H34 or 3a3b can lead to death in mice infected with the influenza A virus. FEMS Immunology & Medical Microbiology, 29(3), 177–181. https://doi.org/10.1111/j.1574-695X.2000
Hasan, H. A., Vijayakumarz H. D., Alias, J., Ismail, N. I., & Kurniawan, S. B. (2025). Treatment and recovery of biosolids from sewage wastewater using bioflocculants Bacillus velezensis isolate JB7. In B. K. Taseli & U. Iyer-Raniga (Eds.) Sewage Management and Treatment Techniques. InTech Open. https://doi.org/10.5772/intechopen.1009764
Ibrahim, M., Griko, N., Junker, M., & Bulla, L. (2010). Bacillus thuringiensis: A genomics and proteomics perspective. Bioengineered Bugs, 1(1), 31–50. https://doi.org/10.4161/bbug.1.1.10519
Kabir, M., Riko, Y. Y., Abdullahi, B., Kabir, K., Zubairu, U. D., & Hamza, U. A. (2020). Bioburdens of selected ready-to-eat fruits and vegetables consumed in Katsina metropolis, Katsina state, Nigeria. International Journal of Science and Research (IJSR), 9(9).
Land, M., Bundschuh, M., Hopkins, R. J., Poulin, B., & McKie, B. G. (2023). Effects of mosquito control using the microbial agent Bacillus thuringiensis israelensis (Bti) on aquatic and terrestrial ecosystems: A systematic review. Environmental Evidence, 12(1). https://doi.org/10.1186/s13750-023-00319-w
Liu, X., Ruan, L., Peng, D. li, L., Sun, M., & Yu, Z. (2014). Thuringiensin: a thermostable secondary metabolite from Bacillus thuringiensis with insecticidal activity against a wide range of insects. Toxins, 6(8), 2229-2238. https://doi.org/10.3390/toxins60822229
Ma, X., Hu, J., Ding, C., Portieles, R., Xu, H., Gao, J., Du, L., Gao, X., Yue, Q., Zhao, L., & Borrás-Hidalgo, O. (2023). New native Bacillus thuringiensis strains induce high insecticidal action against Culex pipiens pallens larvae and adults. BMC Microbiology, 23(1). https://doi.org/10.1186/s12866-023-02842-9
Mahuku, G. S. (2004). A simple extraction method suitable for PCR-based analysis of plant, fungal, and bacterial DNA. Plant Molecular Biology Reporter, 22(1), 71–81. https://doi.org/10.1007/BF02773351
Masri, M. M., Tan, J. S., & Ariff, A. B. (2020). Effect of Cry+ Bacillus thuringiensis cells and fermentation condition on consistent production of δ-endotoxin. *Asia-Pacific Journal of Science and Technology, 25*(02), APST-25. https://so01.tci-thaijo.org/index.php/APST/article/view/185274
McKie, B. G., Taylor, A., Nilsson, T., Frainer, A., & Goedkoop, W. (2023). Ecological effects of mosquito control with Bti: Evidence for shifts in the trophic structure of soil- and ground-based food webs. Aquatic Sciences, 85, 47. https://doi.org/10.1007/s00027-023-00944-0
Milne, R. Y., Gauthier, D., & Van Frankenhuyzen, K. (2008). Purification of Vip3Aa from Bacillus thuringiensis HD-1 and its contributions to toxicity of HD-1 to spruce budworm (Choristoneura fumiferana) and gypsy moth (Lymantria dispar) (Lepidoptera). Journal of Invertebrate Pathology, 99, 166–172. https://doi.org/10.1016/j.jip.2008.05.002
Monnerat, R. G., Dias, D. G. S., Da Silva, S. F., Martins, E. S., Berry, C., Falcão, R., Gomes, A. C. M. M., Praca, L. B., & Soares, C. M. S. (2005). Screening of Bacillus thuringiensis strains effective against mosquitoes. Pesquisa Agropecuaria Brasileira, 40(2), 103–106. https://doi.org/10.1590/S0100-204X2005000200001
Nazina, T., Tourova, T., Poltaraus, A., Novikova, E., Grigoryan, A., Ivanova, A., Lysenko, A., Petrunya, V., Osipov, G., Belyaev, S., & Iyanov, M. (2001). Taxonomic study of aerobic thermophilic Bacilli. International Journal of Systematic and Evolutionary Microbiology, 51(2), 433–446. https://doi.org/10.1099/00207713-51-2-433
Pigott, C. R., King, M. S., & Ellar, D. J. (2008). Investigating the properties of Bacillus thuringiensis Cry proteins with novel loop replacements created using combinatorial molecular biology. Applied and Environmental Microbiology, 74(11), 3497–3511. https://doi.org/10.1128/AEM.02844-07
Patel, M., Raymond, B., Bonsall, M. B., & West, S. A. (2019). Crystal toxins and the volunteer's dilemma in bacteria. Journal of Evolutionary Biology, 32(4), 310–319. https://doi.org/10.1111/jeb.13415
Poopathi, S., & Archana, B. (2012). Optimization of medium composition for the production of mosquitocidal toxins from Bacillus thuringiensis subsp. israelensis. Indian Journal of Experimental Biology, 50(1), 65–71.
Shankar, K., Prabakaran, G., & Am, M. (2016). Cost-effective medium for the production of mosquitocidal toxins from a novel strain Bacillus. Insect Biochemistry and Molecular Biology, 41(7), 42.
Toledo, F., Calvo, C., Rodelas, B., & Gonzalez-Lopez, J. (2006). Selection and identification of bacteria isolated from waste crude oil with polycyclic aromatic hydrocarbons removal capacities. Systematic and Applied Microbiology, 29(3), 244–252. https://doi.org/10.1016/j.syapm.2005.09.003
Trisyono, Y. A., Aryuwandari, V. E. F., Rahayu, T., Martinelli, S., Head, G. P., Parimi, S., & Camacho, L. R. (2023). Baseline susceptibility of the field populations of Ostrinia furnacalis in Indonesia to the proteins Cry1A.105 and Cry2Ab2 of Bacillus thuringiensis. Toxins, 15(10), 602. https://doi.org/10.3390/toxins15100602
Viljoen, C. D., Booysen, C., & Tantuan, S. S. (2022). The suitability of using spectrophotometry to determine the concentration and purity of DNA extracted from processed food matrices. Journal of Food Composition and Analysis, 112, 104689. https://doi.org/10.1016/j.jfca.2022.104689
Wei, J. Z., Hale, K., Carta, L., Platzer, E., Wong, C., Fang, S. C., & Aroian, R. (2003). Bacillus thuringiensis crystal proteins that target nematodes. Proceedings of the National Academy of Sciences, 100, 2760–2765. https://doi.org/10.1073/pnas.0538072100
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Ummulkhairi Saulawa Aminu, Zubairu Darma Umar, Muhammad Sani Aliyu

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.