Phenotypic and Genotypic Analysis of Haemolysin Genes of Staphylococcus aureus Isolated from Subclinical Mastitis in Savojbolagh County, Alborz Province

Document Type : Full Research Paper

Authors

1 Department of Biology, Faculty of Basic Sciences, Borujerd Branch, Islamic Azad University, Borujerd, Iran.

2 Department of Pathobiology, School of Veterinary Medicine, Shiraz, Iran.

3 Department of Microbiology, Faculty of Veterinary Medicine, Karaj Branch, Islamic Azad University, Karaj, Iran.

4 Depatment of Virology, Razi vaccine and Serum Research Institute, Shiraz, Iran.

5 Department of Microbiology, Islamic Azad University, Borujerd Branch, Borujerd, Iran.

Abstract

Staphylococcus aureus pathogens are known in animals and humans. They cause various infections such as mastitis in cattle, sheep and goats, as well as dermatitis, poisoning and even in some cases severe infections such as toxic shock syndrome (TSS) in humans. The bacteria also lead to severe economic losses and mortality in dairy industry, due to the fact that the bacteria can become resistant to antibiotics as a result of treatment makes it difficult. Studies have shown that there is a relationship between mastitis and bacterial virulence factors, one of which is haemolysin enzyme. The aim of this study was to analyze the phenotypes and genotypes of haemolysin genes in Staphylococcus aureus isolated from 540 subclinical bovine mastitis in Savojbolagh county, Alborz province. Subclinical mastitis-affected quarters which has been detected by CMT, were cultured in the laboratory. Staphylococcus aureus samples positive for coagulase test were analyzed by PCR and the results of PCR and culture were compared. Of 420 subclinical bovine mastitis samples, 84(20%) were determined positive by culture. Fifty (59.53%) samples were found positive by coagulase test. Forty five samples (90%) out of this 50 were positive by PCR test. These 45 samples were culture on blood agar and analyzed by PCR . The results showed that in some cases, based on bacterial cultures, haemolysin virulence genes were well expressed and the results almost collapsed with the results of PCR. Hence, we can say that in most cases, there is a significant relationship (P value = 0/001) between the phenotypes and genotypes of virulence genes. However, there is no direct relation, because in some cases, genes are not expressed in culture as well and this is quite obvious in alpha haemolysin gene.

Keywords

References

1- Ebrahimi, A. Akhavan Taheri, M., (2009),Characteristics of staphylococci isolated from clinical and subclinical mastitis cows in Shahrekord, Iran, Iranian Journal of Veterinary Research, Shiraz University, Vol. 10, No. 3, Ser. No. 28.
2- Bolourchi M, Mokhber Dezfouli M. R., Kasravi R., Moghimi Esfandabadi A., Hovareshti, P. (2008),An estimation of national average of milk somatic cell count and production losses due to subclinical mastitis in commercial dairy herds in Iran. J Vet. Res., 63: 263-266.
3- Watanabe S, Ito T, Sasaki T, Li S, Uchiyama I, Kishii K, Kikuchi K, Leo Skov R, Hiramatsu K (2009),Genetic Diversity of Staphylocoagulase Genes (coa): Insight into the Evolution of Variable Chromosomal Virulence Factors in Staphylococcus aureus, journal.pone DOI: 10.1371.
4- Fei w, hongjun y , bin h, changfa w, yundong g , qifeng zh(2011),study on the haemolysin phenotype and the genotype distribution of staphylococcus aureus caused bovine mastitis in Shandong dairy farms, Intern J Appl Res Vet Med, vol. 9, NO:4.
5- Bhakdi ,S, Tranum, J(1991),Alpha-toxin of Staphylococcus aureus. J, microbial. Rev, 55(4) 733-751.
6- Julio C. Franco G, Libertad González V, Sandra C. Gómez M, Juan M. Carrillo G. and José J. Ramírez C(2008), Virulence factors analysis of Staphylococcus aureus isolated from bovine mastitis in México, J, eGnosis,Vol. 6, Art. 7.
7- Saumyas S,srivasta va,satyabrata P,Amita S,Neesar A,Aejazur R, Krishnasastry V.membrane (2009), bound monomer of staphylococcal a-haemolysin induces caspase activation and apoptotic cell death despite initiation of membrane repair pathway, J. plos one 4(7):e6293
8- Füssle R, Bhakdi S, Sziegoleit A, Tranum-Jensen J, Kranz T, Wellensiek HJ(1981).on the mechanism of membrane damage by Staphylococcus aureus alpha toxin, ,J.cell Biol,91(1),83-9
9- Cifrian E, Guidry A,J, Bramley A, J (1996), Effect of staphylococcal beta toxin on the cytotoxcity ,proliferation and adherence of Staphylococcus aureus to bovine mammary epithelial cell. J.vet microbial, 48(3/4),187-198.
10- Ariyanti, D, Salasia, S, Tato S, (2011) Characterization of Haemolysin of Staphylococcus aureus Isolated from Food of Animal Origin, Journal of Biotechnology, Vol. 16, No:1 PP:32.37.
11- Delgado S, GarcIa P, Fernandez L, Jimenez E, RodrIguez-Banos M, del Campo R & Juan M, (2011), Characterization of Staphylococcus aureus strains involved in Human and bovine mastitis, J, FEMS Immunol Med Microbiol 62 ,225–235.
12- Rusenova N,Gebreyes W, Koleva M, Mitev J, Penev T, Vasilev N, Mitev A T(2013), Comparison of Three Methods for Routine Detection of Staphylococcus aureus Isolated from Bovine Mastitis, J ,Kafkas Univ Vet Fak Derg, 19 (4): 709-712.
13- Momtaz H, Rahimi E , Tajbakhsh E (2010), Detection of some virulence factors in Staphylococcus aureus isolated from clinical and subclinical bovine mastitis in Iran, African Journal of Biotechnology Vol. 9(25), pp. 3753-3758.
14- Da Silva, R. E., Boechat, J. U. D., Martins, J. C. D., Ferreira, W. P. B., Sequera, A. P. S., and Da silva, N (2005), Haemolysin Production by Staphylococcus aureus Species Isolated from Mastitic Goat Milk in Brazilian Dairy Herd. J. Science. Small rum. res., 56, 271-275.
15- Straub, Joachim A.; Hertel, Christian; Hammes, Walter P, A (1999),23S rDNA-Targeted Polymerase Chain Reaction–Based System for Detection of Staphylococcus aureus in Meat Starter Cultures and Dairy Products, Journal of Food Protection. pp. 1103-1227, pp. 1150-1156(7).
16- Kenny K, Reiser R F, Bastida-Corcuera F D and Norcross N L (1993), Production of enterotoxins and toxic shock syndrome toxin by bovine mammary isolates of Staphylococcus aureus, March J. Clin. Microbiol. vol. 31 no. 3 706-707.
Veterinary Research & Biological Products
Volume 30, Issue 1
March 2017
Pages 14-20

Files

History

  • Receive Date: 20 December 2015
  • Revise Date: 26 January 2016
  • Accept Date: 17 April 2016

Share

How to cite

Statistics