New strains of Bacillus thuringiensis subsp. israelensis highly toxic for Aedes aegypti and Culex pipiens pipiens

Cover Page

Cite item

Full Text

Open Access Open Access
Restricted Access Access granted
Restricted Access Subscription Access

Abstract

Bacillus thuringiensis subsp. israelensis (Bti) is a known subspicies of crystal-forming entomopathogenic bacteria used to control blood-sucking mosquitoes. In this work, we isolated three different strains of Bti 4369, 4929 and 4999 from the wild larvae midgut of blood-sucking mosquitoes Aedes flavescens . The bacterial isolates were identified by the 16S rRNA gene and serotype determination revealed that the strains belonged to Bt subsp. israelensis H14. The strains had differences in bacterial colony morphology, a number of biochemical characteristics and protein endotoxin profiles. The isolated strains Bti 4369 and 4999 showed high insecticidal activity against Culex pipiens pipiens and Aedes aegypti larvae, with LC50 values of 1.47 ×108 ‒2.26 × 108 spores/ml 24 hours after treatment. The value for strain Bti 4929 LC50 was 32.7‒35.9 × 108 spores/ml. The new isolated strains of Bacillus thuringiensis subsp. israelensis have high potential for the development of ecological friendly bioinsecticides for the control of blood-sucking mosquitoes.

Full Text

Restricted Access

About the authors

V. P. Khodyrev

Institute of Systematics and Ecology of Animals SB RAS

Email: ovp0408@yandex.ru
Russian Federation, Novosibirsk, 630091

O. V. Polenogova

Institute of Systematics and Ecology of Animals SB RAS

Author for correspondence.
Email: ovp0408@yandex.ru
Russian Federation, Novosibirsk, 630091

A. S. Artemchenko

Institute of Systematics and Ecology of Animals SB RAS

Email: ovp0408@yandex.ru
Russian Federation, Novosibirsk, 630091

A. V. Krivopalov

Institute of Systematics and Ecology of Animals SB RAS

Email: ovp0408@yandex.ru
Russian Federation, Novosibirsk, 630091

V. V. Glupov

Institute of Systematics and Ecology of Animals SB RAS

Email: ovp0408@yandex.ru
Russian Federation, Novosibirsk, 630091

References

  1. Гаджиева С. С. Филогенетическая структура и состав фауны кровососущих комаров ( Diptera , Culicidae ) Северного Кавказа и факторы, определяющие их динамику // Известия ДГПУ. Естественные и точные науки. 2021. Т. 15. № 1. С. 27–32. https://doi.org/10.31161/1995-0675-2021-15-1-27-32
  2. Мирзаева А. Г., Смирнова Ю. А., Юрченко Ю. А., Кононова Ю. А. К познанию фауны и экологии кровососущих комаров ( Diptera , Culicidae ) лесостепных и степных районов Западной Сибири // Паразитология. 2007. Т. 41. № 4. С. 253‒267.
  3. Халин А. В., Айбулатов С. В., Филоненко И. В. Распространение кровососущих комаров ( Diptera , Culicidae ) на северо-западе России: виды рода Aedes meigen // Энтомол. обозр. 2021. T. 100. С. 755‒796. https://doi.org/10.31857/S0367144521040055
  4. Baig D. N., Mehnaz S. Determination, and distribution of cry-type genes in halophilc Bacillus thuringiensis isolates of Arabian Sea sedimentary rocks // Microbiol. Res. 2010. V. 165. P. 376–383. https://doi.org/10.1016/j.micres.2009.08.003
  5. Becker N. Ice granules containing endotoxins of microbial agents for the control of mosquito larvae ‒ a new application technique // J. Am. Mosq. Control Assoc. 2003. V. 19. P. 63–66.
  6. Ben-Dov E., Zaritsky A., Dahan E., Barak Z., Sinai R., Manasherob R., Khamraev A., Troitskaya E., Dubitsky A., Berezina N., Margalith Y. Extended screening by PCR for seven cry-group genes from field-collected strains of Bacillus thuringiensis // Appl. Environ. Microbiol. 1997. V. 63. P. 4883–4890. https://doi.org/10.1128/aem.63.12.4883-4890.1997
  7. Ben-Dov E. Bacillus thuringiensis subsp. israelensis and its dipteran-specific toxins // Toxins. 2014. V. 6. P. 222–1243. https://doi.org/10.3390/toxins6041222
  8. Bravo A., Gill S. S., Soberón M. Mode of action of Bacillus thuringiensis toxins and their potential for insect control // Toxicon. 2007. V. 49. P. 423–435. https://doi.org/10.1016/j.toxicon.2006.11.022
  9. Bravo A., Likitvivatanavong S., Gill S. S., Soberón M. Bacillus thuringiensis : a story of a successful bioinsecticide // Insect Biochem. Mol. Biol. 2011. V. 41. P. 423–431. https://doi.org/10.1016/j.ibmb.2011.02.006
  10. Canton P. E., Reyes E. Z., de Escudero I. R., Bravo A., Soberon M. Binding of Bacillus thuringiensis subsp. israelensis Cry4Ba to Cyt1Aa has an important role in synergism // Peptides. 2011. V. 32. P. 595–600.
  11. Chertkova E., Kabilov M. R., Yaroslavtseva O., Polenogova O., Kosman E., Sidorenko D., Alikina T., Noskov Y., Krivopalov A., Glupov V. V., Kryukov V.Yu. Links between soil bacteriobiomes and fungistasis toward fungi infecting the colorado potato beetle // Microorganisms. 2023. V. 11. Art. 943. https://doi.org/10.3390/microorganisms11040943
  12. Crickmore N., Bone E. J., Willians J. A., Ellar D. J. Contribution of the individual components of the δ-endotoxin crystal to the mosquitocidal activity of Bacillus thuringiensis subsp. israelensis // FEMS Microbiol. Lett. 1995. V. 131. P. 249–254.
  13. De Barjac H., Bonnefoi A. Essai de classification biochimique et sérologique de 24 souches de Bacillus de type B. thuringiensis // Entomophaga. 1962. V. 1. P. 5–31.
  14. De Barjac H., Bonnefoi A. Mise au point sur la classification des Bacillus thuringiensis // Entomophaga. 1973. V. 18. P. 5–17.
  15. De Maagd R. A., Bravo A., Crickmore N. How Bacillus thuringiensis has evolved specific toxins to colonize the insect world // Trends Genet. 2001. V. 17. P. 193–199. https://doi.org/10.1016/S0168-9525(01)02237-5
  16. De Maagd R. A., Bravo A., Berry C., Crickmore N., Schnepf H. E. Structure, diversity, and evolution of protein toxins from spore-forming entomopathogenic bacteria // Annu. Rev. Genet. 2003. V. 37. P. 409–433. https://doi.org/10.1146/annurev.genet.37.110801.143042
  17. Federici B. A., Lüthy P., Ibarra J. E. Parasporal body of Bacillus thuringiensis israelensis : structure, protein composition, and toxicity // Bacterial control of mosquitoes & black flies. Biochemistry, genetics & applications of Bacillus thuringiensis israelensis and Bacillus sphaericus / Eds. H. de Barjac, D.J. Sutherland. London, UK: Unwin Hyman, 1990. P. 16–44.
  18. Federici B. A., Park H.-W., Sakano Y. Insecticidal Protein Crystals of Bacillus thuringiensis // Inclusions in Prokaryotes/ Ed. Shively J. M. Springer-Verlag, Berlin, Heidelberg, 2006. Р. 195-235. https://doi.org/10.1007/3-540-33774-1_8
  19. Fernández-Chapa D., Ramírez-Villalobos J., Galán-Wong L. Toxic potential of Bacillus thuringiensis : an overview. Protecting rice grains in the post-genomic era. IntechOpen, 2019. https://doi.org/10.5772/intechopen.85756
  20. González-Villarreal S.E., García-Montelongo M., Ibarra J. E. Insecticidal activity of a Cry1Ca toxin of Bacillus thuringiensis Berliner ( Firmicutes : Bacillaceae ) and its synergism with the Cyt1Aa toxin against Aedes aegypti ( Diptera : Culicidae ) // J. Med. Entomol. 2020. V. 57. P. 1852–1856.
  21. Höfte H., Whiteley H. R. Insecticidal crystal proteins of Bacillus thuringiensis // Microbiol. Rev. 1989. V. 53. P. 242–255. https://doi.org/10.1128/mr.53.2.242-255.1989
  22. Ibarra J. E., del Rincón M. C., Ordúz S., Noriega D., Benintende G., Monnerat R., Regis L., de Oliveira C. M., Lanz H., Rodriguez M. H., Sánchez J., Peña G., Bravo A. Diversity of Bacillus thuringiensis strains from Latin America with insecticidal activity against different mosquito species / / Appl. Environ. Microbiol. 2003. V. 69. P. 5269–5274. https://doi.org/10.1128/AEM.69.9.5269-5274
  23. Ishii T., Ohba M. The 23-kilodalton CytB protein is solely responsible for mosquito larvicidal activity of Bacillus thuringiensis serovar kyushuensis // Curr. Microbiol. 1994. V. 29. P. 91–94. https://doi.org/10.1007/BF01575754
  24. García-Suárez R., Verduzco-Rosas L. A, Ibarra J. E. Isolation and characterization of two highly insecticidal, endophytic strains of Bacillus thuringiensis // FEMS Microbiol. Ecol. 2021. V. 97. Art. fiab080. https://doi.org/10.1093/femsec/fiab080
  25. Kumar P., Kamle M., Borah R., Kumar D. K., Sharma B. Bacillus thuringiensis as microbial biopesticide: uses and application for sustainable agriculture // Egypt. J. Biol. Pest Control. 2021. V. 31. Art. 95. https://doi.org/10.1186/s41938-021-00440-3
  26. Lai L., Villanueva M., Muruzabal-Galarza A., Fernández A. B., Unzue A., Toledo-Arana A., Caballero P., Caballero C. J. Bacillus thuringiensis Cyt proteins as enablers of activity of Cry and Tpp toxins against Aedes albopictus // Toxins. 2023. V. 15. Art. 211. https://doi.org/10.3390/toxins15030211
  27. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage // Nature. 1970. V. 227. P. 680–685.
  28. Liu H. M., Yang P. P., Cheng P., Wang H. F., Liu L. J., Huang X., Zhao Y. Q., Wang H. W., Zhang C. X., Gong M. Q. Resistance level of mosquito species ( Diptera : Culicidae ) from Shandong Province, China // Int. J. Insect Sci. 2015. V. 7. P. 47–52. https://doi.org/10.4137/IJIS.S24232
  29. Ma X., Hu J., Ding C., Portieles R., Xu H., Gao J., Du L., Gao X., Yue Q., Zhao L., Borrás-Hidalgo O. New native Bacillus thuringiensis strains induce high insecticidal action against Culex pipiens pallens larvae and adults // BMC Microbiol. 2023. V. 23. P. 100. https://doi.org/10.1186/s12866-023-02842-9
  30. Manasherob R., Itsko M., Sela-Baranes N., Ben-Dov E., Berry C., Cohen S., Zaritsky A. Cyt1 Ca from Bacillus thuringiensis subsp. israelensis : production in Escherichia coli and comparison of its biological activities with those of other Cyt-like proteins // Microbiology (Reading). 2006. V. 152. P. 2651–2659.
  31. McClintock J.T., Schaffer C. R., Sjoblad R. D. A comparative review of the mammalian toxicity of Bacillus thuringiensis -based pesticides // Pestic. Sci. 1995. V. 45. P. 95–105.
  32. Mishra P. K., Bisht S. C., Ruwari P., Subbanna A. R.N.S., Bisht K., Bhatt J., Gupta H. S. Genetic diversity and functional characterization of endophytic Bacillus thuringiensis isolates from the North Western Indian Himalayas // Ann. Microbiol. 2017. V. 67. P. 143–155. https://doi.org/10.1007/s13213-016-1244-0
  33. Onen H., Luzala M. M., Kigozi S., Sikumbili R. M., Muanga C.-J.K., Zola E. N., Wendji S. N., Buya A. B., Bal ciunaitiene A., Viškelis J., Kaddumukasa M. A., Memvanga P. B. Mosquito-borne diseases and their control strategies: an overview focused on green synthesized plant-based metallic nanoparticles // Insects. 2023. V. 14. Art. 221. https://doi.org/10.3390/insects14030221
  34. Orduz S., Diaz T., Restrepo N., Patiño M. M., Tamayo M. C. Biochemical, immunological and toxicological characteristics of the crystal proteins of Bacillus thuringiensis subsp. medellin // Mem. Inst. Oswaldo Cruz. 1996. V. 91. P. 231–237. https://doi.org/10.1590/S0074-02761996000200020
  35. Pérez C., Fernandez L. E., Sun J., Folch J. L., Gill S. S., Soberón M., Bravo A. Bacillus thuringiensis subsp. israelensis Cyt1Aa synergizes Cry11Aa toxin by functioning as a membrane-bound receptor // Proc. Nat. Acad. Sci. USA. 2005. V. 102. P. 18303‒18308.
  36. Raymond B., Johnston P. R., Nielsen-LeRoux C., Lereclus D., Crickmore N. Bacillus thuringiensis : An impotent pathogen? // Trends Microbiol. 2010. V. 18. P. 189–194. https://doi.org/10.1016/j.tim.2010.02.006
  37. Reyes-Ramirez A., Ibarra J. E. Fingerprinting of Bacillus thuringiensis type strains and isolates by using Bacillus cereus group specific repetitive extragenic palindromic sequence-based PCR analysis // Appl. Environ. Microbiol. 2005. V. 71. P. 1346–1355. https://doi.org/10.1128/AEM.71.3.1346-1355.2005
  38. Schnepf E., Crickmore N., Van Rie J., Lereclus D., Baum J., Feitelson J., Zeigler D. R., Dean D. H. Bacillus thuringiensis and its pesticidal crystal proteins // Microbiol. Mol. Biol. Rev. 1998. V. 62. P. 775–806. https://doi.org/10.1128/MMBR.62.3.775-806.1998
  39. Shishir A., Roy A., Islam N., Rahman A., Khan S. N., Hoq M. M. Abundance, and diversity of Bacillus thuringiensis in Bangladesh and their cry genes profile // Front. Environ. Sci. 2014. V. 2. Art. 20. https://doi.org/0.3389/fenvs.2014.00020
  40. Soares-da-Silva J., Queirós S. G., de Aguiar J. S., Viana J. L., Neta M. D.R.A.V., da Silva M. C., Pinheiro V. C.S., Polanczyk R. A., Carvalho-Zilse G.A., Tadei W. P. Molecular characterization of the gene profile of Bacillus thuringiensis Berliner isolated from brazilian ecosystems and showing pathogenic activity against mosquito larvae of medical importance // Acta Trop. 2017. V. 176. P. 197–205. https://doi.org/10.1016/j.actatropica.2017.08.006
  41. Sur B., Nigam N., Joshi A. K., Bihari V. Characterization of mosquito larvicidal Bacillus thuringiensis isolated from soils of India // Indian J. Biotechnol. 2003. V. 2. P. 268–270.
  42. Valtierra-de-Luis D., Villanueva M., Lai L., Williams T., Caballero P. Potential of Cry10Aa and Cyt2Ba, two minority δ-endotoxins produced by Bacillus thuringiensis ser. israelensis , for the control of Aedes aegypti larvae // Toxins. 2020. V. 12. Art. 355.
  43. Van Frankenhuyzen K . Cross-order and cross-phylum activity of Bacillus thuringiensis pesticidal proteins // J. Invert. Pathol. 2013. V. 114. P. 76–85. https://doi.org/10.1016/j.jip.2013.05.010
  44. Yan M., Roehrl M. H., Wang J. Y. Discovery of crystalline inclusions in Bacillus licheniformis that resemble parasporal crystals of Bacillus thuringiensis // Can. J. Microbiol. 2007. V. 53. P. 1111‒1115. https://doi.org/10.1139/W07-076
  45. Weisburg W. G., Barns S. M., Pelletier D. A., Lane D. J. 16s ribosomal DNA amplification for phylogenetic study // J. Bacteriol. 1991. V. 173. P. 697–703.
  46. Wu D., Cheng F. N. Synergism in mosquitocidal activity of 26 and 65 kDa proteins from Bacillus thuringiensis subsp. israelensis crystal // FEBS Lett. 1985. V. 190. P. 232–236. https://doi.org/10.1016/0014-5793(85)81290-4

Supplementary files

Supplementary Files
Action
1. JATS XML
Download
2. Fig. 1. Comparative morphological assessment of 6-day bacterial colonies of Bt subsp. israelensis isolated from the midgut of Aedes flavescens larvae; a ‒ 4369; b ‒ 4929; c ‒ 4999.

Download (23KB)
Indexing metadata
3. Fig. 2. Microscopic analysis (×100, Axsioscop 40) of 6-day bacterial colonies of Bt subsp. israelensis isolated from the midgut of Aedes flavescens larvae; a ‒ 4369; b ‒ 4929; c ‒ 4999.

Download (28KB)
Indexing metadata
4. Fig. 3. Comparative electrophoresis of spore-crystal mixtures of the studied strains: 1 – 4369; 2 – 4929; 3 – 4999. M – pre-stained molecular weight standards (8‒200 kDa) (“Servicebio, China”).

Download (26KB)
Indexing metadata

Copyright (c) 2024 Russian Academy of Sciences