Investigation of genotypic diversity of Paenibacillus larvae strains isolated from Iranian honey bee colonies.

Document Type : Full Research Paper

Authors

1 Department of Honey Bee, Silk Worm a Wildlife, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran

2 Department of Microbial Biotechnology, Faculty of Basic Sciences, University of Science and Culture, Tehran, Iran

Abstract

American Foulbrood disease is a deadly bacterial disease of honey bee larvae in the world. The disease is caused by gram-positive and spores forming bacteria of Paenibacillus larvae that has significant economic losses. However, no study has been done on the diversity of this disease agent in Iran. In this study, 19 strains of Penibacillus larvae that were isolated from diseased and dead larvae and honey after confirmation by biochemical tests, PCR of 16S rRNA gene and sequencing, were used in ERIC-PCR technique to study genotypic diversity. The results enabled the identification of ERIC I and ERIC II genotypes for the first time in Iran.  14 strains of genotype II (73.7%) and 5 strains showed genotype I (26.3%).

Keywords

References

1. Alippi, A. M., A. C. Lopez, F. J. Reynaldi, D. H. Grasso and O. M. Aguilar. 2007. Evidence for plasmid-mediated tetracycline resistance in Paenibacillus larvae, the causal agent of American Foulbrood (AFB) disease in honey bees. Veterinary Microbiology 125 (3) 4: 290-303.
2. Bassi, S., G. Formato, M. Milito, K. Trevisiol, C. Salogni and E. Carra. 2014. Phenotypic characterization and ERIC-PCR based genotyping of Paenibacillus larvae isolates recovered from American foulbrood outbreaks in honey bees from Italy. Veterinary Quarterly, Vol. 35, pp. 27-32.  
3. Beims, H., B.‌ Bunk, S. Erler, K. I. Mohr, C. Spröer, S. Pradella, G. Günther, M. Rohde, W. v. d. Ohe and M. ‌‌Steinert. 2020. Discovery of Paenibacillus larva ERIC V: Phenotypic and genomic comparison to genotypes ERIC I-IV reveal different inventories of virulence factors which correlate with epidemiological prevalences of American Foulbrood. International Journal of Medical Microbiology 310: 151394.
4. De Graaf, D. C., A. M. Alippi, K. Antunez, K. A. Aronstein, G. Budge, D. De Koker, L. De Smet, D. W. Dingman, J. D. Evans, L. J. Foster, A. Funfhaus, E. Garcia-Gonzalez, A. Gregorc, H. Human, K. D. Murray, B. K. Nguyen, L. Poppinga, M. Spivak, D. Van Engelsdorp, S. Wilkins and E. Genersch. 2013. Review Article: Standard methods for American foulbrood research. Journal of Apicultural Research 52. 
5. Di Pinto, A., L. Novello, V. Terio and G. Tantillo. 2011. ERIC-PCR Genotyping of Paenibacillus larvae in Southern Italian Honey and Brood Combs. Current Microbiology 63: 416–419.
6. ‌Djukic, M., E. Brzuszkiewicz, A. Fünfhaus, J. Voss, K. Gollnow, L. Poppinga, H. Liesegang, E. Garcia-Gonzalez, E. Genersch and R. Daniel. 2014. How to kill the honey bee larva: genomic potential and virulence mechanisms of Paenibacillus larvae. PLOS ONE, Vol. 9, Issue. 3.
7. Ebeling, J., H. Knispel, G. Hertlein, A. Fünfhaus and E. Genersch. 2016. Biology of Paenibacillus larvae, a deadly pathogen of honey bee larvae. Applied Microbiology and Biotechnology 100: 7387–7395. 
8. Fünfhaus, A., L. Poppinga and E. Genersch. 2013. Identification and characterization of two novel toxins expressed by the lethal honey bee pathogen Paenibacillus larvae, the causative agent of American foulbrood. Environmental Microbiology 15: 2951–2965.
9. Genersch, E., A. Ashiralieva and I. Fries. 2005. ‌strain and genotype-specific differences in virulence of Paenibacillus larvae subsp. larvae, a bcterial pathogen causing American foulbrood disease in honey bees‌. Applied and environmental microbiology, Vol. 71, no. 11, pp. 7551–7555.
10. ‌‌‌Genersch, E., E. Forsgren, J. Pentikainen, A. Ashiralieva, S. Rauch, J. Kilwinski and ‌I. Fries. 2006. Reclassification of Paenibacillus larvae subsp. Pulvifaciens and Paenibacillus larvae subsp. larvae as Paenibacillus larvae without subspecies differentiation. International Journal of Systematic and Evolutionary Microbiology 56: 501–511. 
11. Genersch, E. 2010. American Foulbrood in honey bees and its causative agent, Paenibacillus larvae. The Journal of Invertebrate Pathology 103: 10–19.
12. Grady, E. N., J. MacDonald and L. Liu. 2016. Current knowledge and perspectives of Paenibacillus: a review. Microbial Cell Factories 15: 203.
13. Heyndrickx, M., K. Vandemeulebroecke and B. Hoste. 1996. Reclassification of Paenibacillus pulvifaciens as a subspecies of P. larvae, with emended descriptions of P. larvae as P. larvae subsp. Larvae and P. larvae subsp. pulvifaciens. International Journal of Systematic Bacteriology 46: 270–279.
14. Liang, T. W and S. L. Wang. 2015. Recent advances in exopolysaccharides from Paenibacillus spp: production, isolation, structure, and bioactivities. Mar Drugs 13: 1847–1863.
15. OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. 2018. American foulbrood of the honey bee. Chapter 3.2.2.
16. Piccini, C., B. D'Alessandro, K. Antunez and P. Zunino. 2002. Detection of Paenibacillus larvae subsp. Larvae spores in naturally infected bee larvae and artificially contaminated honey by PCR. The World Journal of Microbiology & Biotechnology 18: 761-765. 
17. ‌Rusenova, N., P. Parvanov and S. Stanilova. 2013. Molecular Typing of Paenibacillus larvae Strains Isolated from Bulgarian Apiaries Based on Repetitive Element Polymerase Chain‌ ‌ ‌‌‌‌Reaction (Rep-PCR). Current Microbiology.   
18 Versalovic, J., M. Schneider, F. J. De Bruijn and J. R. Lupski. 1994. Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction. Methods in molecular cell biology 5: 25-40. 
19. ‌Yuko, H., S. Toshinari, I. Nanami, M. Naokazu and T. Daisuke. 2016. Existence of Paenibacillus larvae genotypes ERIC I-ST2, ERIC I-ST15 and ERIC II-ST10 in the western region of Aichi prefecture, Japan. Journal of Veterinary Medical Science 78 (7): 1195-1199. 
 20. Yousten, A. A and J. L. Capinera. 2018. Encyclopedia of entomology. Dordrecht Springer, pp. 2718–2719. 
 
Veterinary Research & Biological Products
Volume 35, Issue 4 - Serial Number 4
December 2022
Pages 91-97

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History

  • Receive Date: 19 January 2022
  • Accept Date: 12 February 2022

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