Virulence gene profile and antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis in Kashmir, India
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Abstract
The Staphylococcus aureusis often responsible for a number of diseases in humans and animals, and it is considered as a main etiological agent of bovine mastitis. The pathogenicity of S. aureus is due to both its ability to resist antibiotics, and the production of toxins. This study investigated virulence genes, prevalence and antibiogram profile of S. aureus isolated from dairy cows suffering from mastitis in the Kashmir. A total of 70 S. aureus isolates were obtained from 250 mastitic milk samples collected from both organized (47/150) and unorganized (23/100) dairy farms. Five pathogenic factors including clfA, hld, seo, lukM, and coa and one resistance gene mecAgene were checked through PCR. Clumping factor gene (clfA) was found in most of the isolates with a percentage of 81.42 % whereas,hld, seo, lukM, and coa were present in 61.2, 54.28, 70, and 71 percent of isolates, respectively. However, amplification of coagene yielded DNA bands of two different sizes. A high percentage of antimicrobial resistance rates were observed, wherein, Ampicillin showed highest resistance with 85.7 %, followed by Kanamycin, Cefotaxime, Sulphadizine and Streptomicin showing 71.42 %, 54.28 %, 51.48 % and 42 %, respectively. A high frequency of Methicillin resistant S. aureus (MRSA 28.57 %) was observed in these isolates and all methicillin resistant isolates were found to be positive for mecA gene via PCR amplification. These results revealed that mastitis-associated S. aureus among bovines of Kashmir is able to accumulate different virulence factors and resistance to antimicrobials, making the treatment of infections difficult.
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Keywords
Antibiogram, Genotypic characterization, Mastitis, PCR, Staphylococcus aureus
References
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Barrio, M. B., Rainard, P. and Prevost, G. (2006). LukM/LukF'-PV is the most active Staphylococcus aureus leukotoxin on bovine neutrophils. Microbes and Infection.,8: 2068-74
Booth, M. C., Pence, L. M., Mahasreshti, P., Callegan, M., and Gilmore, M. (2001). Clonal associations among Staphylococcus aureus isolates from various sites of infections. Infection and Immunity, 69: 345-352
Botrel, M. A., Haenni, M., Morignat, E., Sulpice., Madec, J. Y. and Calavas, D. (2010). Distribution and antimicrobial resistance of clinical and subclinical mastitis pathogens in dairy cows in Rhône-Alpes, France. Food borne Pathogens and Disease, 7: 479-487
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
CLSI. Performance Standards for Antimicrobial Disk and Dilution Susceptibility for Bacteria Isolated from Animals; Approved Standard - Second Edition. CLSI document M31-A3,28: 65–72. Wayne, PA, USA: Clinical and Laboratory Standards Institute, 2008.
Dego, K., Vandijk, J. E. and Nederbragt, H. (2002). Factors involved in the early pathogenises of Bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion. Veterinary Quarterly, 24: 181-198
Dhanda, M. R. and Sethi, M. S. (1962): Investigation of Mastitis in India.Icar Res. Series 35, New Delhi, India.
Dinges, M. M, Orwin, P. M. and Schlievert, P. M. (2000). Exotoxins of Staphylococcus aureus.Journal of Clinical Microbiology,13: 16-34
Dua, K. (2001): Incidence, etiology and estimated economic losses due to mastitis in Punjab and in India-An update. Indian Dairyman, 53: 41-48
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 aureusfrom birds. Journal of veterinary medicine.B, Infectious Diseases and Veterinary Public Health, 52: 38-44
Fueyo, J. M., Mendoza, M. C., Rodicio., Muniz, M. R. J., Alvarez, M. A. and . Martín, M. C. (2005).Cytotoxin and Pyrogenic Toxin Superantigen Gene Profiles of Staphylococcus aureus Associated with Subclinical Mastitis in Dairy Cows and Relationships with Macro restriction Genomic Profiles. Journal of Clinical Microbiology, 43: 1278–1284.
Gitau, G. K., Wabacha, J. K., Mulei, C., Mndurumo, S. and Nduhiu, J. M. (2011). Isolation rates and antimicrobial sensitivity patterns of bovine mastitis pathogens in peri-urban area of Nairobi, Kabete, Kenya.Ethiopian Veterinary Journal, 15: 1-13
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Guler, L., Ok, U., Gunduz, K., Gulcu, Y. and Hadimli, H. S. (2005). Antimicrobial susceptibility and coagulase gene typing of Staphylococcus aureus isolated from bovine clinical mastitis cases in Turkey. Journal of Dairy Science, 88: 3149-3154
Haran, K. P., Godden, S. M., Boxrud, D., Jawahir, S. Bender, J. B. and Sreevatsan, S. (2012). Prevalence and Characterization of Staphylococcus aureus, Including Methicillin-Resistant Staphylococcus aureus, Isolated from Bulk Tank Milk from Minnesota Dairy Farms. Journal of Clinical Microbiology, 50: 688-695
Haveri, M., Roslof, A., Rantala, L. and Pyarala, S. (2007). Virulence genes of bovine Staphylococcus aureus from persistent and non-persistent intramammary infections with different clinical characteristics. Journal of Applied Microbiology, 103:993–1000
Horodyska, M. M., Kubiak, J., Lassa, H. and Malinowski, E. (2012). Changes in antibiotic susceptibility pattern of atypical Staphylococcus aureus strains isolated from cows of the same herd in 2008-2010. Bulletin of the Veterinary Institute in Pulawy, 56: 139-14
Hussain, A., Shakoor, A., Yousaf, A., Rehman, S. U. and Zaman, M. A. (2012). Clinical and subclinical Staphylococcus auerus in dairy buffaloes: disease characteristics and antibiotic susceptibility profiles of isolates. The Journal of Animal and Plant Sciences, 22:
217-220
Kalorey, D. R., Shanmugam, Y., Kurkure, N. K., 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
Khakpoor, M., Safarmashaei, S. and Jafary, R. (2011). Study of Milk Extracted from Cows Related to Staphylococcus aureus by Culturing and PCR. Global Veterinaria,7: 572-575
Kumar, R., Yadav, B. R. and Singh, R. S. (2010). Genetic Determinants of antibiotic resistance in Staphylococcus aureus isolates from milk of mastitic crossbred cattle. Current Microbiology, 60: 379-386
Kuzma, K., Malinowski, E., Lassa, H. and K?ossowska, A. (2003). Specific detection of Staphylococcus aureus by PCR in intramammary infection.Bulletin of the Veterinary Institute in Pulawy, 47: 183-190
Lee, J. H. (2003). Methicillin (oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans. Applied and Environmental Microbiology, 69: 6489–6494
Li, J., Zhou, H., Yuan, L., He, T. and Hu, S. (2009). Prevalence, genetic diversity, and antimicrobial susceptibility profiles of Staphylococcus aureus isolated from bovine mastitis in Zhejiang Province, China. Journal of Zhejiang University-Science B,10:753-760
Malinowski, E., Lassa, H., Smulski, S., K?ossowska, A. and Kaczmarowski, M. (2008). Antimicrobial susceptibility of bacteria isolated from cows with mastitis in 2006–2007.Bulletin of the Veterinary Institute in Pulawy, 52: 565–572
Marconi, C., Cunha, M. L. R. S., Araujo, J. P., Jr. and Rugolo, L. M. S. S. (2005). Standardization of the PCR technique for the detection of delta toxin in Staphylococcus spp. Journal of Venomous Animals and Toxins Including Tropical Diseases, 11:117–128
Momtaz, H., Tajbakhsh, E., Rahimi, E. and Momeni, M. (2011). Coagulase gene polymorphism of Staphylococcus aureus isolated from clinical and sub-clinical bovine mastitis in Isfahan and Chaharmahalva Bakhtiari provinces of Iran. Comparative Clinical Pathology, 20:519-522
NDDB, (2012). National Dairy Development Board (NDDB). Annual Report, 2011-12.
Omoe, K., Ishikawa, M., Shimoda, Y., Hu, D. L., Ueda, S. and Shinagawa, K. (2002). Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureusisolates harboring seg, seh, or sei genes. Journal of Clinical Microbiology, 40:857–862
Ote, I., Taminiau, B., Duprez, J.N. Dizier, I. andMainil, J. G. (2011). Genotypic characterization by polymerase chain reaction of Staphylococcus aureus isolates associated with bovine mastitis. Veterinary Microbiology, 153: 285-292
Ranjan, R., Gupta, M. K. and Singh, K. K. (2011). Study of bovine mastitis in different climatic conditions in Jharkhand, India. Veterinary World, 4: 205-208.
Turutoglu, H., Ercelik, S. and Ozturk, D. (2006). Antibiotic resistance of Staphylococcus aureusand Coagulase negative Staphylococci isolated from bovine mastitis. Bulletin of the Veterinary Institute in Pulawy, 50:41–45
Wang, G. Y. and Niu, Z. N. (2009). Isolation and identification of pathogenic bacteria to clinical mastitis in liaochen city.Journal of Shandong Agriculture University
Yamagishi, N., Jinkawa, Y., Omoe, K., Makino, S. and Oboshi, K. (2007). Sensitive test for screening for Staphylococcus aureus in bovine mastitis by broth cultivation and PCR. Veterinary Record, 16: 381-383
Barrio, M. B., Rainard, P. and Prevost, G. (2006). LukM/LukF'-PV is the most active Staphylococcus aureus leukotoxin on bovine neutrophils. Microbes and Infection.,8: 2068-74
Booth, M. C., Pence, L. M., Mahasreshti, P., Callegan, M., and Gilmore, M. (2001). Clonal associations among Staphylococcus aureus isolates from various sites of infections. Infection and Immunity, 69: 345-352
Botrel, M. A., Haenni, M., Morignat, E., Sulpice., Madec, J. Y. and Calavas, D. (2010). Distribution and antimicrobial resistance of clinical and subclinical mastitis pathogens in dairy cows in Rhône-Alpes, France. Food borne Pathogens and Disease, 7: 479-487
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
CLSI. Performance Standards for Antimicrobial Disk and Dilution Susceptibility for Bacteria Isolated from Animals; Approved Standard - Second Edition. CLSI document M31-A3,28: 65–72. Wayne, PA, USA: Clinical and Laboratory Standards Institute, 2008.
Dego, K., Vandijk, J. E. and Nederbragt, H. (2002). Factors involved in the early pathogenises of Bovine Staphylococcus aureus mastitis with emphasis on bacterial adhesion and invasion. Veterinary Quarterly, 24: 181-198
Dhanda, M. R. and Sethi, M. S. (1962): Investigation of Mastitis in India.Icar Res. Series 35, New Delhi, India.
Dinges, M. M, Orwin, P. M. and Schlievert, P. M. (2000). Exotoxins of Staphylococcus aureus.Journal of Clinical Microbiology,13: 16-34
Dua, K. (2001): Incidence, etiology and estimated economic losses due to mastitis in Punjab and in India-An update. Indian Dairyman, 53: 41-48
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 aureusfrom birds. Journal of veterinary medicine.B, Infectious Diseases and Veterinary Public Health, 52: 38-44
Fueyo, J. M., Mendoza, M. C., Rodicio., Muniz, M. R. J., Alvarez, M. A. and . Martín, M. C. (2005).Cytotoxin and Pyrogenic Toxin Superantigen Gene Profiles of Staphylococcus aureus Associated with Subclinical Mastitis in Dairy Cows and Relationships with Macro restriction Genomic Profiles. Journal of Clinical Microbiology, 43: 1278–1284.
Gitau, G. K., Wabacha, J. K., Mulei, C., Mndurumo, S. and Nduhiu, J. M. (2011). Isolation rates and antimicrobial sensitivity patterns of bovine mastitis pathogens in peri-urban area of Nairobi, Kabete, Kenya.Ethiopian Veterinary Journal, 15: 1-13
Goh, S. H., Byrne, S. K., Zhang, J. L. and Chow, A. W. (1992). Molecular typing of Staphylococcus aureus on the basis of coagulase gene polymorphisms. Journal of Clinical Microbiology, 30: 1642-1645
Guler, L., Ok, U., Gunduz, K., Gulcu, Y. and Hadimli, H. S. (2005). Antimicrobial susceptibility and coagulase gene typing of Staphylococcus aureus isolated from bovine clinical mastitis cases in Turkey. Journal of Dairy Science, 88: 3149-3154
Haran, K. P., Godden, S. M., Boxrud, D., Jawahir, S. Bender, J. B. and Sreevatsan, S. (2012). Prevalence and Characterization of Staphylococcus aureus, Including Methicillin-Resistant Staphylococcus aureus, Isolated from Bulk Tank Milk from Minnesota Dairy Farms. Journal of Clinical Microbiology, 50: 688-695
Haveri, M., Roslof, A., Rantala, L. and Pyarala, S. (2007). Virulence genes of bovine Staphylococcus aureus from persistent and non-persistent intramammary infections with different clinical characteristics. Journal of Applied Microbiology, 103:993–1000
Horodyska, M. M., Kubiak, J., Lassa, H. and Malinowski, E. (2012). Changes in antibiotic susceptibility pattern of atypical Staphylococcus aureus strains isolated from cows of the same herd in 2008-2010. Bulletin of the Veterinary Institute in Pulawy, 56: 139-14
Hussain, A., Shakoor, A., Yousaf, A., Rehman, S. U. and Zaman, M. A. (2012). Clinical and subclinical Staphylococcus auerus in dairy buffaloes: disease characteristics and antibiotic susceptibility profiles of isolates. The Journal of Animal and Plant Sciences, 22:
217-220
Kalorey, D. R., Shanmugam, Y., Kurkure, N. K., 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
Khakpoor, M., Safarmashaei, S. and Jafary, R. (2011). Study of Milk Extracted from Cows Related to Staphylococcus aureus by Culturing and PCR. Global Veterinaria,7: 572-575
Kumar, R., Yadav, B. R. and Singh, R. S. (2010). Genetic Determinants of antibiotic resistance in Staphylococcus aureus isolates from milk of mastitic crossbred cattle. Current Microbiology, 60: 379-386
Kuzma, K., Malinowski, E., Lassa, H. and K?ossowska, A. (2003). Specific detection of Staphylococcus aureus by PCR in intramammary infection.Bulletin of the Veterinary Institute in Pulawy, 47: 183-190
Lee, J. H. (2003). Methicillin (oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans. Applied and Environmental Microbiology, 69: 6489–6494
Li, J., Zhou, H., Yuan, L., He, T. and Hu, S. (2009). Prevalence, genetic diversity, and antimicrobial susceptibility profiles of Staphylococcus aureus isolated from bovine mastitis in Zhejiang Province, China. Journal of Zhejiang University-Science B,10:753-760
Malinowski, E., Lassa, H., Smulski, S., K?ossowska, A. and Kaczmarowski, M. (2008). Antimicrobial susceptibility of bacteria isolated from cows with mastitis in 2006–2007.Bulletin of the Veterinary Institute in Pulawy, 52: 565–572
Marconi, C., Cunha, M. L. R. S., Araujo, J. P., Jr. and Rugolo, L. M. S. S. (2005). Standardization of the PCR technique for the detection of delta toxin in Staphylococcus spp. Journal of Venomous Animals and Toxins Including Tropical Diseases, 11:117–128
Momtaz, H., Tajbakhsh, E., Rahimi, E. and Momeni, M. (2011). Coagulase gene polymorphism of Staphylococcus aureus isolated from clinical and sub-clinical bovine mastitis in Isfahan and Chaharmahalva Bakhtiari provinces of Iran. Comparative Clinical Pathology, 20:519-522
NDDB, (2012). National Dairy Development Board (NDDB). Annual Report, 2011-12.
Omoe, K., Ishikawa, M., Shimoda, Y., Hu, D. L., Ueda, S. and Shinagawa, K. (2002). Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureusisolates harboring seg, seh, or sei genes. Journal of Clinical Microbiology, 40:857–862
Ote, I., Taminiau, B., Duprez, J.N. Dizier, I. andMainil, J. G. (2011). Genotypic characterization by polymerase chain reaction of Staphylococcus aureus isolates associated with bovine mastitis. Veterinary Microbiology, 153: 285-292
Ranjan, R., Gupta, M. K. and Singh, K. K. (2011). Study of bovine mastitis in different climatic conditions in Jharkhand, India. Veterinary World, 4: 205-208.
Turutoglu, H., Ercelik, S. and Ozturk, D. (2006). Antibiotic resistance of Staphylococcus aureusand Coagulase negative Staphylococci isolated from bovine mastitis. Bulletin of the Veterinary Institute in Pulawy, 50:41–45
Wang, G. Y. and Niu, Z. N. (2009). Isolation and identification of pathogenic bacteria to clinical mastitis in liaochen city.Journal of Shandong Agriculture University
Yamagishi, N., Jinkawa, Y., Omoe, K., Makino, S. and Oboshi, K. (2007). Sensitive test for screening for Staphylococcus aureus in bovine mastitis by broth cultivation and PCR. Veterinary Record, 16: 381-383
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Virulence gene profile and antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis in Kashmir, India. (2017). Journal of Applied and Natural Science, 9(2), 893-898. https://doi.org/10.31018/jans.v9i2.1294
