Identification and Typing of Staphylococcus aureus Isolated from Clinical Bovine Mastitis in Sanandaj City Using PCR-RFLP Analysis of the aroA Gene

Document Type : Original Article

Authors

10.35066/J040.2018.526

Abstract

Staphylococcus aureus is one of the most important causes of contagious mastitis in domestic animals. Due to the existence of multiple strains of S. aureus and strain variations in chromosomal allelic rearrangement, different genotyping methods, such as analysis of the chromosomal DNA following the enzymatic digestion, are introduced for genetic typing of the bacterium. In the present survey, for the first time, the genetic diversity of S. aureus recovered from clinical bovine mastitis in Sanandaj was investigated based on PCR-RFLP analysis of the aroA gene. 120 bovine mastitic milk samples were collected aseptically and assessed. S. aureus isolated in culture and routine bacteriological methods were analyzed by aroA gene-based PCR. The amplicons with a size of 1153bp were digested with TaqI restriction enzyme and the fragments were electrophoresed. 28 S. aureus strains were isolated in phenotypic method among which the expected amplicon was observed in all of them. In enzymatic digestion, two RFLP patterns, nomenclatural based on the previous studies were generated. Genotype B was detected in 23 (82.14%) isolates and genotype N in 5 isolates (17.85%). The results demonstrate that S. aureus is involved in bovine mastitis in Sanandaj and despite the presence of limited genotypes, strain variation of this bacterium exist in the region. The presence of genotype B in all farms implies the same source of infection among farms, which should be considered in control programs of mastitis. Non-detection of universal genotype A and specific genotype of the bacterium in Sanandaj is reported for the first time.

Keywords

References

1- Scherrer D, Corti S, Muehlherr JE, Zweifel C, Stephan R. Phenotypic and genotypic characteristic of Staphylococcus aureus isolates from raw bulk-tank milk samples of goat sand sheep. Vet Microbiol. 2004; 101(1):101-107.
2- El-Seedy FR, El-Shabrawy M, Hakim AS, Dorgham SM, Ata SN, Bakry MA, et al. Recent techniques used for isolation and characterization of Staphylococcus aureus from mastitic cows. J Am Sci 2010; 6(12):701-708.
3- Güler L, Ok Ü, Gündüz K, Gülcü Y, Hadimli HH. Antimicrobial susceptibility and coagulase gene typing of Staphylococcus aureus isolated from bovine clinical mastitis cases in Turkey. J Dairy Sci. 2005; 88:3149-3154.
4- Cascon AS, Anguita JC, Hernanz CM, Sa´nchez MS, Yugueros JM, Naharro GC. RFLP-PCR analysis of the aroA gene as a taxonomic tool for the genus Aeromonas. FEMS Microbiol Lett. 1997; 156:199-204.
5- Marcos JY, Soriano AC, Sa´nchez MS, Moral CH, Ramos SS, Smeltzer MS, et al. Rapid identification and typing of Staphylococcus aureus by PCR-restriction fragment length polymorphism analysis of the aroA gene. J Clin Microbiol. 1999; 37:570-574.
6- EL-Huneidi W, Bdour S, Mauasnen A. Detection of enterotoxin genes seg, seh, sei and sej and of a novel aroA in Jordanian clinical isolates of staphylococcus aureus.
Diagn Microbiol Infect Dis. 2006; 56:127-132.
7- Dastmalchi HS, Ahmadi M, Mardani K, Batavani RA. Genotyping of Staphylococcus aureus isolated from bovine mastitis based on PCR-RFLP analysis of the aroA gene.
Comp Clin Path. 2010; 19:163-168.
8- Talebi-Satlou R, Ahmadi M, Ghavam F, Dastmalchi HS. Use of 5- enolpyruvylshikmate-3phosphate synthase encoding gene for typing of Staphylococcus aureus isolated from skin and urinary tract Infections of human. Iran J Basic Med Sci. 2012; 15:975-982.
9- Hope AF. Laboratory handbook on bovine mastitis. 1st ed. Madison, Wisconsin: National Mastitis Council (NMC); 1999. p. 113-123.
10- Quinn PJ, Markey Bk, Carter ME, Donnelly WJC, Leonard FC, Maguire D. Veterinary Microbiology and Microbial Diseases.  1st Ed. New York: Blackwell Science; 2002. p. 315-328.
11- Bhati T, Nathawat P, Sharma SK, Yadav R, Bishnoi J, Kataria AK. Polymorphism in spa gene of Staphylococcus aureus from bovine subclinical mastitis. Vet World. 2016; 9(4):421-424.
12- Bendahou A, Lebbadi M, Ennanei L, Essadqui F, Abid M. Characterization of Staphylococcus species isolated from raw milk and milk products in North Morocco.
J Infect Dev Ctries. 2008; 2:218-225.
13- Sasidharan S, Prema B, Yoga LL. Antimicrobial drug resistance of Staphylococcus aureus in dairy products. Asian Pac J Trop Biomed. 2011; 1(2):130-132.
14- Van Leeuwen WB, Melles DC, Alaidan A, Al-Ahdal M, Boelens HAM, Snijders SV, et al. Host and tissue-specific pathogenic traits of staphylococcus aureus.       J Bacteriol. 2005; 187:4584-4591.
15- De Oliveira LP, Soares e Barros LS, Silva VC, Cirqueira MG. Study of Staphylococcus aureus in raw and pasteurized milk consumed in the Reconcavo area of the State of Bahia, Brazil. Journal of Food Processing and Technology. 2011; 2(6):128-132.
16- Huijps K, Lam TJ, Hogeveen H. Costs of mastitis: facts and perception. J Dairy Res. 2008; 75:113-120.
New Findings in Veterinary Microbiology
Volume 1, Issue 2
April 2019
Pages 26-35

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History

  • Receive Date: 15 December 2017
  • Revise Date: 04 January 2018
  • Accept Date: 03 March 2018

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