Physical Properties of Bacillus Toxins and Their Larvicidal Activity Against Anopheles Arabiensis Mosquito Larvae

Authors

  • Manhal Ahmed. Hamza Omdurman Islamic University
  • Sulieman Mohamed El-Sanousi University of Khartoum, Khartoum

DOI:

https://doi.org/10.47941/ijbs.1012

Keywords:

Bacillus thuringiensis, Bacillus sphaericus, toxins, Physical properties, larvicidal activity, mosquito larvae.

Abstract

Purpose: Biological control has been strongly encouraged by using of entomopathoginic bacteria Bacillus thuringiensis and Bacillus sphaericus. The present study monitored the larvicidal effect of Bacillus thuringiesis and Bacillus sphaericus against Anopheles mosquito larvae and the effect of physical factors on the larvicidal potency on mosquito larvae.

Methodology: Mid-guts of Anopheles mosquito larvae were inoculated on both Blood Agar and MacConkey Agar culture media. Depending on Gram stain to determine gram positive and gram-negative bacteria, and to identify the bacterial cells` morphology. A 1000µl of Bacillus thuringiensis. Sample was heated at 80°C for 12 min and diluted from 10 -1 to l0-8 with sterile normal saline. Bacillus strains were inoculated into Müller Hinton broth media and incubated over-night at 37°C. This time was quite enough to form the spores and crystals. The supernatant fluid was discarded and the cell suspensions were washed with sterile distilled water the process was repeated up to 3 times until spores and crystals were free from debris by using phase contrast microscopy. Collected spores and crystals were kept in sterile containers in fridge until used.

Results: Fifty-three mid-gut contents of Anopheles arabiensis larvae were identified morphologically and biochemically. Bacillus species were the most prevalent bacteria. The highest larval mortality rate was recorded by 5ml of Bt1 and Bt2 after 24 hrs.  of treatment and by 150µl of Bacillus species spore.100% mortality was recorded by the Bacillus spore-crystal mixture. A mortality of 60% was recorded at pH 8.0, by 400 C the mortality rate was 20%at 24 hrs. After 12 hours, the exposure to UV light, larvicidal activity reduced to 40 %. 

Unique Contribution to Theory Policy and Practices: These findings revealed that both B.thuringiensis and Bacillus sphaericus are a good control agent for Anopheles mosquitoes. More deep investigations are needed to verify the effect of the entomopathogens, which has biological and economic importance in mosquito control. In addition, Intensive studies must be needed to validate reliability.

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Author Biographies

Manhal Ahmed. Hamza, Omdurman Islamic University

Faculty of Medical Laboratory Sciences, Department of Medical Microbiology

Sulieman Mohamed El-Sanousi, University of Khartoum, Khartoum

Faculty of Veterinary medicine

References

Abdel Rahman., I. M. , 2005.Laboratory Trials for Chemical and Biological Control of the Mosquitoes Anopheles arabiensis and Culex quinquefasciatus (Diptera: Culicidae). M. Sc. Thesis. U. of K.

Ahmed, N.A.M. (2006).The effect of Bacillus larvicidal toxin on Mammals. M. Sc. Thesis. U. of K.

Barrow, G.I. and Feltham,R.K.A.(1993).Cowan and Steel's Manual for the Identification of Medical Bacteria . 3rd ed.Cambridge University Press, Cambridge, UK.pp.51, 86-90.

Balakrishnan.S.,Indira.K. and Srinivasan.M. Mosquitocidal properties of Bacillus species isolated from mangroves of Vellar estuary, Southeast coast of India. J Parasit Dis. 2015 Sep; 39(3): 385-392.

Bravo. A., Gill. S.S., Soberón. M. Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon 2007; 49: 423-435.

Brown MD, Watson TM, Carter J, Purdie DM, Kay BH. Toxicity of VectoLex (Bacillus sphaericus) products to selected Australian mosquito and nontarget species. J Econ Entomol 2004; 97:51-8.

Capalbo, D.M.F.; Vilas-Bôas, G.T.; Suzuki, M.T.; Arantes, O.M.N. Bacillus thuringiensis: formulações e plantas transgênicas. Biotecnolog. Cienc. Desenvolv, 34, 76-83, 2005.

Carrozi, N. P.; V. C. Kramer; G. W. Warren; S. Evols; and M. G. Kozeil. 1991. Prediction of insecticidal activity of isolates of Bacillus thuringiensis strains by polymerase chain reaction product profiles. Appl. Environ. Microbiol. 57: 3057-3061.Tortora et al. 1998).

Charles JF, Nielsen-LeRoux C, Delcluse A. Bacillus sphaericus toxins: molecular biology and mode of action. Annu Rev Entomol. 1996; 41:451-472.

Crickmore, N., Bone, E. J Williams, J. A and Ellar. D. J Contribution of the individual components of the δ--endotoxin crystal to mosquitocidal activity of Bacillus thuringiensis subsp. israelensis. FEMS Microbiol. Lett. 1995.131: 249 Ã254.

Crickmore. N., Zeigler. D.R, Feitelson. J, et al. Revision of the nomenclature of the Bacillus thuringiensis pesticidal crystal proteins. Microbiol Mol Biol Rev 1998; 62: 807-813.

Cruickshank,R.;Duguid,J.;Marmion,B.P.and Swain,R.H.(1975).Medical Microbiology.12th.ed. Vol.2. London and NewYork.

Darboux, I., C. Nielsen-LeRoux, J.-F. Charles, and D. Pauron. 2001. The receptor of Bacillus sphaericus binary toxin in Culex pipiens (Diptera: Culicidae) midgut: molecular cloning and expression. Insect Biochem. Mol. Biol. 31: 981Ã990.

Das. P.K., Amalraj D.D. 1997. Biological control of malarial vectors. Indian Journal of Medicine, 106: 174-197.

Davidson, E. W. 1995. Biochemistry and mode of action of the Bacillus sphaericus toxins. Mem. Inst. Oswaldo Cruz 90: 81Ã 86

ElÒ«in, Y.M.(1995). Control of mosquito larvae by encapsulated Pathogen Bacillus thuringiensis var.israelensis.J.Microencapsulation,12(5).515-523.

Federici, BA., Park, HW., Bideshi, DK., Wirth, MC., Johnson, JJ., Sakano, Y., Tang, M. Developing recombinant bacteria for control of mosquito larvae. In: Floore, TG., editor.

Biorational Control of Mosquitoes. American Mosquito Control Association; Mount Laurel, NJ: 2007. p. 164-175.

Gorashi , N.E. Elshafie, H.A.F,Hamid,H. A. and Dirar. H. D. Characterization of Sudan strains of Bacillus thuringiensis pathogenic to the larvae of the house mosquito Culex quinquefasciatus. Agric. Biol. J. N. Am., 2012, 3(7): 271-279

Jahn, A., Yong, S., Choi, J. and Kim, D. (2010). A bioassay for mosquito repellency against Aedes aegypti: method validation and bioactivities of DEET analogues, 91-97.

Lacey, L.A.; Lacey, C.M. The medicinal importance of riceland mosquitoes and their control using alternatives to chemical insecticides. J. Am. Mosq. Control Assoc. 1990, 2, 1-93.

Larget-Thiery, I. (1984). Simulation studies on the persistence of Bacillus thuringiensis H - 14 > WHO/VBC/84.906.J Parasit Dis. 2015 Sep; 39(3): 385-392.

Maniafu, B. M., Wilber, L., Ndiege, I. O., Wanjala, C. C. and Akenga, T. A. (2009). Larvicidal activity of extracts from three Plumbago spp against Anopheles gambiae. Memorias Do Instituto Oswaldo Cruz, 104(6), 813-817.

Mittal , P . K . ( 2003 ) Biolarvicides in vector control: challenges and prospects . Journal of Vector Borne Diseases , 40 , 20 - 32 .

Mulligan, F.S.; Schaefer, C.H., and Wilder, W.H. (1980). Efficacy and persistence of Bacillus sphaericus and B. thuringiensis H - 14 against mosquitoes under laboratory and field conditions. J. econ. Entomol. 73, 684 - 688.

Murray, R. G. and C. F. Robinow. 1981. Schaeffer Fulton method for staining endospores. p. 17-33. In: Gerhardt, P,; R, G. E. Murray; R. N. Castilow; E. W. Nester; W. A. Wood; N. R. Krieg and G. B. Phillips (eds). Manual of Methods for General Bacteriology. American Scociety for Microbiology.

Nugud,A.D.(1982). Towards to Control of Anopheles arabiensis, Patton:evaluation of Bacillus thuringiensis Berliner(H-14) and other biological agents affecting mosquitoes .PhD thesis, University of London.

Omer. S. M. 1990. Bacteria Naturally parasitising mosquitoes at Khartoum and the possible larvicidal agent. Ph.D. Thesis (U.K).

Omoya, F. O. and Boboye, B. E. Influence of Physicochemical Parameters on Mosquito Larvicidal Potency of Bacillus subtilis Isolated from Musca domestica (Linn) Cadavers in Nigeria. MRJI, 2017; 19(6): 1-8,; MRJI.31806.

Park HW, Bideshi DK, Federici BA. The 20-kDa protein of Bacillus thuringiensis subsp. israelensis enhances Bacillus sphaericus 2362 Bin toxin synthesis. Curr Microbiol. 2007a; 55:119-124. [PubMed: 17597341.

Park HW, Federici BA. Genetic engineering of bacteria to improve efficacy using the insecticidal proteins of Bacillus species. In: Stock SP, editor. Insect pathogens: molecular approach and techniques. Cambridge: CABI International; 2009. pp. 275-305

Rosouvitz,MJ. et al(1998). In: Microbiology and Microbial Infections. Vol:2 9 th . Edited by Topley and Wilson's Systematic Bicteriology. Great Britain. Arnold, London.

Sneath, P. H. A. 1986. Endospore-forming gram positive Rods and Cocci. p. 1104-1105 In: Sneath, P. H. A.; Mour, N. S.; Sharpe, M. E. and Hoit, J. G. (eds). Bergy's Manual of Systematic Bacteriology, 8 th edition. Vol. 2, William & Wilkins. Baltimore.

Soad,O.M.(1900) Bacteria naturally parasitizing mosquitoes of Khartoum as possible larvicidal agents.Ph.D. Thesis. University of Khartoum.

Travers, R.S.; Martin, P.A.W.; Reichelderfer, C.F. Selective Process for Efficient Isolation of Soil Bacillus spp. Appl. Environ. Microbiol. 1987, 53, 1263-1266.

Wirth, M.C.; Park, H.-W.; Walton, W.E.; Federici, B.A. Cyt1A of Bacillus thuringiensis delays evolution of resistance to Cry11A in the mosquito Culex quinquefasciatus. Appl. Environ. Microbial. 2005, 71, 185-189.

WHO1, (2019) Guidelines for malaria vector control. World Health Organization.

Yousef. N. S., Aly. N. S. M Effectiveness of Bacillus thuringiensis and Bacillus sphaericus isolates against Culex pipens, Aedes egypti and Anopheles sergenti Mosquito Larvae. PUJ; 2013, 6(1): 1-6.

Zhu YC, Kramer KJ, Oppert B, Dowdy AK. cDNAs of aminopeptidase-like protein genes from Plodia interpunctella strains with different susceptibilities to Bacillus thuringiensis toxins. Insect Biochemistry and Molecular Biology. 2000; 30:215-224.

WHO, (2019). World Malaria Report 2019. World Health Organization-Geneva. Retrieved from https://www.who.int/publications-detail/world-malaria-report-2019

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Published

2022-09-01

How to Cite

Hamza, M. A., & El-Sanousi, S. M. (2022). Physical Properties of Bacillus Toxins and Their Larvicidal Activity Against Anopheles Arabiensis Mosquito Larvae. International Journal of Biological Studies, 2(1), 44–59. https://doi.org/10.47941/ijbs.1012

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