Tropical Animal Health and Production

, Volume 44, Issue 8, pp 1821–1826 | Cite as

Detection of virulence-associated genes in Staphylococcus aureus isolated from bovine clinical mastitis milk samples in Guangxi

  • Feng Li Yang
  • Xiao Shan Li
  • Xian Wei Liang
  • Xiu Fang Zhang
  • Guang Sheng Qin
  • Bing Zhuang Yang
Original Research
First Online:
Accepted:

Abstract

Staphylococcus aureus is recognized worldwide as a pathogen causing many serious diseases in humans and animals and is one of the most common etiological agents of clinical and subclinical bovine mastitis. The purpose of this study was to determine the presence of genes encoding clfA, fnbA, fnbB, cap5, cap8, hla, hlb, nuc, sea, and tst of S. aureus strains (n = 39) isolated from bovine clinical mastitis in Guangxi by polymerase chain reaction amplification. The results of the present study indicated that all isolates were found to contain one or more virulence-associated genes. The most frequently encountered genes were fnbA (97 %) and nuc (90 %), followed by hla (85 %) and hlb (82 %), respectively. None of the investigated S. aureus strains harbored fnbB and sea genes. The data in the present study showed a relatively wide distribution of the genes fnbA and nuc among the investigated isolates, indicating that they play an important role on bovine mastitis pathogenesis. The study provides a valuable insight into the virulence-associated genes of this important pathogen.

Keywords

Bovine mastitis Staphylococcus aureus Virulence-associated genes Polymerase chain reaction (PCR) 

Abbreviations

S. aureus

Staphylococcus aureus

clfA

Clumping factor A

fnbA

Fibronectin-binding protein A

fnbB

Fibronectin-binding protein B

cap5

Capsular polysaccharide 5

cap8

Capsular polysaccharide 8

hla

α-Hemolysin

hlb

β-Hemolysin

nuc

Thermonuclease

sea

Enterotoxin A

tst

Toxic shock syndrome toxin 1

PCR

Polymerase chain reaction

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Notes

Acknowledgments

The authors would like to thank Prof. B. X. He, Prof. X. B. Zheng, and Dr. G. H. Li of Guangxi University for their experimental advices. This work was jointly funded by the “948” Project (Ministry of Agriculture of China, no. 2011-G26) and Guangxi Department of Science and Technology (nos. 1123005-1, 1123005-3, and 2010GXnsFA013096).

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Akineden, O., Annemuller, C., Hassan, A.A., Lammler, C., Wolter, W. and Zschock, M., 2001. Toxin genes and other characteristics of Staphylococcus aureus isolates from milk of cows with mastitis. Clinical and Diagnostic Laboratory Immunology, 8, 959-964.PubMedGoogle Scholar
  2. Aarestrup, F.M., Larsen, H.D., Eriksen, N.H.R., Elsberg, C.S. and Jensen, N.E., 1999. Frequency of alpha- and beta-haemolysin in Staphylococcus aureus of bovine and human origin. A comparison between pheno- and genotype and variation in phenotypic expression. APMIS, 107, 425-430.Google Scholar
  3. Balaban, N. and Rasooly, A., 2000. Staphylococcal enterotoxins. International Journal of Food Microbiology, 61, 1-10.PubMedCrossRefGoogle Scholar
  4. Barkema, H.W., Schukken, Y.H. and Zadoks, R.N., 2006. Invited Review: The role of cow, pathogen, and treatment regimen in the therapeutic success of bovine Staphylococcus aureus mastitis. Journal of Dairy Science, 89, 1877-1895.PubMedCrossRefGoogle Scholar
  5. Brakstad, O.G., Aasbakk, K. and Maeland, J.A., 1992. Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. Journal of Clinical Microbiology, 30, 1654-1660.PubMedGoogle Scholar
  6. Chiang, Y.C., Liao, W.W., Fan, C.M., Pai, W.Y., Chiou, C.S. and Tsen, H.Y., 2008. PCR detection of Staphylococcal enterotoxins (SEs) N, O, P, Q, R, U, and survey of SE types in Staphylococcus aureus isolates from food-poisoning cases in Taiwan. International Journal of Food Microbiology, 121, 66-73.PubMedCrossRefGoogle Scholar
  7. Cifrian, E., Guidry, A.J., Bramley, A.J., Norcross, N.L., Bastida-Corcuera, F.D. and Marquardt, W.W., 1996. Effect of staphylococcal beta toxin on the cytotoxicity, proliferation and adherence of Staphylococcus aureus to bovine mammary epithelial cells. Veterinary Microbiology, 48, 187-198.PubMedCrossRefGoogle Scholar
  8. Cremonesi, P., Luzzana, M., Brasca, M., Morandi, S., Lodi, R., Vimercati, C., Agnellini, D., Caramenti, G., Moroni, P. and Castiglioni, B., 2005. Development of a multiplex PCR assay for the identification of Staphylococcus aureus enterotoxigenic strains isolated from milk and dairy products. Molecular and Cellular Probes, 19, 299-305.PubMedCrossRefGoogle Scholar
  9. Da Silva, E.R., Boechat, J.U.D., Martins, J.C.D., Ferreira, W.P.B., Siqueira, A.P.S. and Da Silva, N., 2005. Hemolysin production by Staphylococcus aureus species isolated from mastitic goat milk in Brazilian dairy herds. Small Ruminant Research, 56, 271-275.CrossRefGoogle Scholar
  10. Dinges, M.M., Orwin, P.M. and Schlievert, P.M., 2000. Exotoxins of Staphylococcus aureus. Clinical Microbiology Reviews, 13, 16-34.PubMedCrossRefGoogle Scholar
  11. El-Sayed, A., Alber, J., Lammler, C., Bonner, B., Huhn, A., Kaleta, E.F. and Zschock, M., 2005. PCR-based detection of genes encoding virulence determinants in Staphylococcus aureus from birds. Journal of veterinary medicine series B, Infectious diseases and veterinary public health, 52, 38-44.CrossRefGoogle Scholar
  12. Fitzgerald, J.R., Hartigan, P.J., Meaney, W.J. and Smyth, C.J., 2000. Molecular population and virulence factor analysis of Staphylococcus aureus from bovine intramammary infection. Journal of Applied Microbiology, 88, 1028-1037.PubMedCrossRefGoogle Scholar
  13. Fowler, T., Wann, E.R., Joh, D., Johansson, S., Foster, T.J. and Hook, M., 2000. Cellular invasion by Staphylococcus aureus involves a fibronectin bridge between the bacterial fibronectin-binding MSCRAMMs and host cell beta1 integrins. European Journal of Cell Biology, 79, 672-679.PubMedCrossRefGoogle Scholar
  14. Getahun, K., Kelay, B., Bekana, M., Lobago. F., 2008. Bovine mastitis and antibiotic resistance patterns in Selalle smallholder dairy farms, central Ethiopia. Tropical Animal Health and Production, 40, 261-268.PubMedCrossRefGoogle Scholar
  15. Guidry, A., Fattom, A., Patel, A., O'Brien, C., Shepherd, S. and Lohuis, J., 1998. Serotyping scheme for Staphylococcus aureus isolated from cows with mastitis. American Journal of Veterinary Research, 59, 1537-1539.PubMedGoogle Scholar
  16. Gunaydin, B., Aslantas, O. and Demir, C., 2011. Detection of superantigenic toxin genes in Staphylococcus aureus strains from subclinical bovine mastitis. Tropical Animal Health and Production, 43, 1633-1637.PubMedCrossRefGoogle Scholar
  17. Johnson, W.M., Tyler, S.D., Ewan, E.P., Ashton, F.E., Pollard, D.R. and Rozee, K.R., 1991. Detection of genes for enterotoxins, exfoliative toxins, and toxic shock syndrome toxin 1 in Staphylococcus aureus by the polymerase chain reaction. Journal of Clinical Microbiology, 29, 426-430.PubMedGoogle Scholar
  18. Jonsson, K., Signas, C., Muller, H.P. and Lindberg, M., 1991. Two different genes encode fibronectin binding proteins in Staphylococcus aureus. The complete nucleotide sequence and characterization of the second gene. European Journal of Biochemistry, 202, 1041-1048.PubMedCrossRefGoogle Scholar
  19. Kalorey, D.R., Shanmugam, Y., Kurkure, N.V., Chousalkar, K.K. and Barbuddhe, S.B., 2007. PCR-based detection of genes encoding virulence determinants in Staphylococcus aureus from bovine subclinical mastitis cases. Journal of Veterinary Science, 8, 151-154.PubMedCrossRefGoogle Scholar
  20. Lachica, R.V., Hoeprich, P.D. and Riemann, H.P., 1972. Tolerance of staphylococcal thermonuclease to stress. Applied microbiology, 23, 994-997.PubMedGoogle Scholar
  21. Larsen, H.D., Aarestrup, F.M. and Jensen, N.E., 2002. Geographical variation in the presence of genes encoding superantigenic exotoxins and beta-hemolysin among Staphylococcus aureus isolated from bovine mastitis in Europe and USA. Veterinary Microbiology, 85, 61-67.PubMedCrossRefGoogle Scholar
  22. Madison, B.M. and Baselski, V.S., 1983. Rapid identification of Staphylococcus aureus in blood cultures by thermonuclease testing. Journal of Clinical Microbiology, 18, 722-724.PubMedGoogle Scholar
  23. Momtaz, H., Rahimi, E. and Tajbakhsh, E., 2010. Detection of some virulence factors in Staphylococcus aureus isolated from clinical and subclinical bovine mastitis in Iran. African Journal of Biotechnology, 25, 3753-3758.Google Scholar
  24. Moore, P.C. and Lindsay, J.A., 2001. Genetic variation among hospital isolates of methicillin-sensitive Staphylococcus aureus: evidence for horizontal transfer of virulence genes. Journal of Clinical Microbiology, 39, 2760-2767.PubMedCrossRefGoogle Scholar
  25. Nashev, D., Toshkova, K., Salasia, S.I., Hassan, A.A., Lammler, C. and Zschock, M., 2004. Distribution of virulence genes of Staphylococcus aureus isolated from stable nasal carriers. FEMS Microbiology Letters, 233, 45-52.PubMedCrossRefGoogle Scholar
  26. Ote, I., Taminiau, B., Duprez, J., Dizier, I. and Mainil, J.G., 2011. Genotypic characterization by polymerase chain reaction of Staphylococcus aureus isolates associated with bovine mastitis. Veterinary Microbiology, 153, 285-292.PubMedCrossRefGoogle Scholar
  27. Peacock, S.J., Moore, C.E., Justice, A., Kantzanou, M., Story, L., Mackie, K., O'Neill, G. and Day, N.P., 2002. Virulent combinations of adhesin and toxin genes in natural populations of Staphylococcus aureus. Infection and Immunity, 70, 4987-4996.PubMedCrossRefGoogle Scholar
  28. Poutrel, B., Boutonnier, A., Sutra, L. and Fournier, J.M., 1988. Prevalence of capsular polysaccharide types 5 and 8 among Staphylococcus aureus isolates from cow, goat, and ewe milk. Journal of Clinical Microbiology, 26, 38-40.PubMedGoogle Scholar
  29. Reinoso, E.B., El-Sayed, A., Lammler, C., Bogni, C. and Zschock, M., 2008. Genotyping of Staphylococcus aureus isolated from humans, bovine subclinical mastitis and food samples in Argentina. Research in Microbiology, 163, 314-322.CrossRefGoogle Scholar
  30. Renzoni, A., Francois, P., Li, D., Kelley, W.L., Lew, D.P., Vaudaux, P. and Schrenzel, J., 2004. Modulation of fibronectin adhesins and other virulence factors in a teicoplanin-resistant derivative of methicillin-resistant Staphylococcus aureus. Antimicrobial Agents and Chemotherapy, 48, 2958-2965.PubMedCrossRefGoogle Scholar
  31. Rich, M., 2005. Staphylococci in animals: prevalence, identification and antimicrobial susceptibility, with an emphasis on methicillin-resistant Staphylococcus aureus. British Journal of Biomedical Science, 62, 98-105.PubMedGoogle Scholar
  32. Roghmann, M., Taylor, K.L., Gupte, A., Zhan, M., Johnson, J.A., Cross, A., Edelman, R. and Fattom, A.I., 2005. Epidemiology of capsular and surface polysaccharide in Staphylococcus aureus infections complicated by bacteraemia. The Journal of Hospital Infection, 59, 27-32.PubMedCrossRefGoogle Scholar
  33. Salasia, S.I., Khusnan, Z., Lammler, C. and Zschock, M., 2004. Comparative studies on pheno- and genotypic properties of Staphylococcus aureus isolated from bovine subclinical mastitis in central Java in Indonesia and Hesse in Germany. Journal of Veterinary Science, 5, 103-109.PubMedGoogle Scholar
  34. Sears, P.M., Gonzalez, R.N., Wilson, D.J. and Han, H.R., 1993. Procedures for mastitis diagnosis and control. The Veterinary Clinics of North America: Food Animal Practice, 9, 445-468.PubMedGoogle Scholar
  35. Sharma, N.K., Rees, C.E. and Dodd, C.E., 2000. Development of a single-reaction multiplex PCR toxin typing assay for Staphylococcus aureus strains. Applied and Environmental Microbiology, 66, 1347-1353.PubMedCrossRefGoogle Scholar
  36. Signas, C., Raucci, G., Jonsson, K., Lindgren, P.E., Anantharamaiah, G.M., Hook, M. and Lindberg, M., 1989. Nucleotide sequence of the gene for a fibronectin-binding protein from Staphylococcus aureus: use of this peptide sequence in the synthesis of biologically active peptides. Proceedings of the National Academy of Sciences of the United States of America, 86, 699-703.PubMedCrossRefGoogle Scholar
  37. Smeltzer, M.S., Gillaspy, A.F., Jr Pratt, F.L., Thames, M.D. and Iandolo, J.J., 1997. Prevalence and chromosomal map location of Staphylococcus aureus adhesin genes. Gene, 196, 249-259.PubMedCrossRefGoogle Scholar
  38. Straub, J.A., Hertel, C. and Hammes, W.P., 1999. A 23S rDNA-targeted polymerase chain reaction-based system for detection of Staphylococcus aureus in meat starter cultures and dairy products. Journal of Food Protection, 62, 1150-1156.PubMedGoogle Scholar
  39. Tamarapu, S., McKillip, J.L. and Drake, M., 2001. Development of a multiplex polymerase chain reaction assay for detection and differentiation of Staphylococcus aureus in dairy products. Journal of Food Protection, 64, 664-668.PubMedGoogle Scholar
  40. Tsen, H.Y. and Chen, T.R., 1992. Use of the polymerase chain reaction for specific detection of type A, D and E enterotoxigenic Staphylococcus aureus in foods. Applied Microbiology and Biotechnology, 37, 685-690.PubMedCrossRefGoogle Scholar
  41. Tuchscherr, L.P., Buzzola, F.R., Alvarez, L.P., Caccuri, R.L., Lee, J.C. and Sordelli, D.O., 2005. Capsule-negative Staphylococcus aureus induces chronic experimental mastitis in mice. Infection and Immunity, 73, 7932-7937.PubMedCrossRefGoogle Scholar
  42. Verdier, I., Durand, G., Bes, M., Taylor, K.L., Lina, G., Vandenesch, F., Fattom, A.I. and Etienne, J., 2007. Identification of the capsular polysaccharides in Staphylococcus aureus clinical isolates by PCR and agglutination tests. Journal of Clinical Microbiology, 45, 725-729.PubMedCrossRefGoogle Scholar
  43. von Eiff, C., Friedrich, A.W., Peters, G. and Becker, K., 2004. Prevalence of genes encoding for members of the staphylococcal leukotoxin family among clinical isolates of Staphylococcus aureus. Diagnostic Microbiology and Infectious Disease, 49, 157-162.CrossRefGoogle Scholar
  44. Wang, S.C., Wu, C.M., Xia, S.C., Qi, Y.H., Xia, L.N. and Shen, J.Z., 2009. Distribution of superantigenic toxin genes in Staphylococcus aureus isolates from milk samples of bovine subclinical mastitis cases in two major diary production regions of China. Veterinary Microbiology, 137, 276-281.PubMedCrossRefGoogle Scholar
  45. Winn, W., Allen, S., Janda, W., Koneman, E., Procop, G., Schreckenberger, P. and Woods, G., 2006. Koneman's Color Atlas and Textbook of Diagnostic Microbiology. Sixth Edition, Lippincott Williams & Wilkins, Philadelphia, pp.623-671.Google Scholar
  46. Xu, S., Arbeit, R.D. and Lee, J.C., 1992. Phagocytic killing of encapsulated and microencapsulated Staphylococcus aureus by human polymorphonuclear leukocytes. Infection and Immunity, 60, 1358-1362.PubMedGoogle Scholar
  47. Yang B., Zhang X.M., Luo S.P. and Liu M.J., 2009. Investigation on Virulence Determinants and Genetic Typing of Staphylococcus aureus Isolated from Mastitis Milk. Acta Agriculturae Boreali-Occidentalis Sinica, 18, 1-6.Google Scholar
  48. Yang, F.L., Li, X.S., He, B.X., Yang, X.L., Li, G.H., Liu, P., Huang, Q.H., Pan, X.M. and Li, J., 2011a. Malondialdehyde level and some enzymatic activities in subclinical mastitis milk. African Journal of Biotechnology, 10, 5534-5538.Google Scholar
  49. Yang, F.L., Li, X.S., He, B.X., Du, Y.L., Li, G.H., Yang, B.B. and Huang, Q.H., 2011b. Bovine Mastitis in Subtropical Dairy Farms, 2005-2009. Journal of Animal and Veterinary Advances, 10, 68-72.CrossRefGoogle Scholar
  50. Yazdankhah, S.P., Solverod, L., Simonsen, S. and Olsen, E., 1999. Development and evaluation of an immunomagnetic separation-ELISA for the detection of Staphylococcus aureus thermostable nuclease in composite milk. Veterinary Microbiology, 67, 113-125.PubMedCrossRefGoogle Scholar
  51. Zecconi, A., Cesaris, L., Liandris, E., Dapra, V. and Piccinini, R., 2006. Role of several Staphylococcus aureus virulence factors on the inflammatory response in bovine mammary gland. Microbial Pathogenesis, 40, 177-183.PubMedCrossRefGoogle Scholar
  52. Zhang, H., Wei, H.W., Cui, Y., Zhao, G.Q. and Feng, F.Q., 2009. Antibacterial interactions of monolaurin with commonly used antimicrobials and food components. Journal of Food Science, 74, 418-421.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Feng Li Yang
    • 1
  • Xiao Shan Li
    • 1
  • Xian Wei Liang
    • 1
  • Xiu Fang Zhang
    • 1
  • Guang Sheng Qin
    • 1
  • Bing Zhuang Yang
    • 1
  1. 1.Guangxi Key Laboratory of Buffalo Genetics and Breeding, Guangxi Buffalo Research InstituteChinese Academy of Agricultural SciencesNanningPeoples Republic of China

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