Comparison of Multilocus Sequence Typing and Spa Typing of Staphylococcus aureus Isolated from Bovine and Bubaline Mastitis
DOI:
https://doi.org/10.48165/ijvsbt.21.5.25Keywords:
Bovine milk, Clonal complexes (CCs), MRSA, Multi-Locus Sequence Typing (MLST), Sequence types (STs)Abstract
Staphylococcus aureus (S. aureus) is the leading causative agent of mastitis in dairy cows, causing reduced milk quality and production. The present study describes the genotypes and clonal distributions of 35 S. aureus strains isolated from cases of mastitis from southern India by multi-locus sequence typing (MLST). The resultant S. aureus sequence types (STs) were compared with their Staphylococcus protein A (spa) types and correlated to methicillin resistance. A total of 27 methicillin-sensitive Staphylococcus aureus (MSSA) and eight methicillin-resistant Staphylococcus aureus (MRSA) strains were identified. Twenty different STs with nine new alleles and nine new STs were obtained. The STs were grouped into four clonal complexes (CCs) CC1, CC5, CC8 and CC97. The predominant STs 2454 and 2459 belonged to CC8 and CC97, respectively. Eight STs were not clustered under any of the CCs. Comparison of genotypes of MLST with that of spa types revealed that some of the isolates with same ST yielded different spa types that were closely related to each other. On the contrary, some of the isolates with same spa types gave different STs in which one ST was the single locus variant of the other in all the cases. Overall, high diversity of S. aureus isolates was observed, excepting for the isolates from a particular geographical region (Bidar) in Karnataka.
Downloads
References
Aires-De-Sousa, M., Boye, K., De Lencastre, H., Deplano, A., et al. (2006). High inter-laboratory reproducibility of DNA sequence-based typing of bacteria in a multicenter study. Journal of Clinical Microbiology, 44, 619-621.
Bardiau, M., Yamazaki, K., Ote, I., Misawa, N., & Mainil, J.G. (2013). Characterization of methicillin-resistant Staphylococcus pseudintermedius isolated from dogs and cats. Microbiology and Immunology, 57(7), 496-501.
Butaye, P., Argudín, M.A., & Smith, T.C. (2016). Livestock-associated MRSA and its current evolution. Current Clinical Microbiology, 3(1), 19-31.
CLSI. (2018). Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals (CLSI Supplement VET08, 4th ed., p. 71). Wayne, PA: Clinical and Laboratory Standards Institute.
Cooper, J.E., & Feil, E.J. (2004). Multilocus sequence typing – what is resolved? Trends in Microbiology, 12(8), 373-377.
Enright, M.C., Day, N.P., Davies, C.E., Peacock, S.J., & Spratt, B.G. (2000). Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus. Journal of Clinical Microbiology, 38(3), 1008-1015.
Gomes, A.R., Westh, H., & De Lencastre, H. (2006). Origins and evolution of methicillin-resistant Staphylococcus aureus clonal lineages. Antimicrobial Agents and Chemotherapy, 50, 3237-3244.
Hallin, M., Deplano, A., Denis, O., De Mendonca, R., De Ryck, R., & Struelens, M.J. (2007). Validation of pulsed-field gel electrophoresis and spa typing for long-term, nationwide epidemiological surveillance studies of Staphylococcus aureus infections. Journal of Clinical Microbiology, 45(1), 127-133.
Haran, K.P., Godden, S.M., Boxrud, D., Jawahir, S., Bender, J.B., & 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.
Hasman, H., Moodley, A., Guardabassi, L., Stegger, M., Skov, R.L., & Aarestrup, F.M. (2010). Spa type distribution in Staphylococcus aureus originating from pigs, cattle and poultry. Veterinary Microbiology, 141, 326-331.
Hata, E., Katsuda, K., Kobayashi, H., Uchida, I., Tanaka, K., & Eguchi, M. (2010). Genetic variation among Staphylococcus aureus strains from bovine milk and their relevance to methicillin-resistant isolates from humans. Journal of Clinical Microbiology, 48, 2130-2139.
Hegde, R. (2011). Rapid identification of bacterial pathogens causing subclinical bovine mastitis with special reference to Staphylococcus aureus, E. coli and predominant streptococcal species by molecular methods (Ph.D. thesis). Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar, India.
Hwang, S.Y., Park, Y.K., Koo, H.C., & Park, Y.H. (2010). Spa typing and enterotoxin gene profile of Staphylococcus aureus isolated from bovine raw milk in Korea. Journal of Veterinary Science, 11(2), 125-131.
Ikawaty, R., Brouwer, E.C., Jansen, M.D., Van Duijkeren, E., Mevius, D., Verhoef, J., & Fluit, A.C. (2009). Characterization of Dutch Staphylococcus aureus from bovine mastitis using a multiple locus variable number tandem repeat analysis. Veterinary Microbiology, 136(3-4), 277-284.
Klein, E., Smith, D.L., & Laxminarayan, R. (2007). Hospitalizations and deaths caused by methicillin-resistant Staphylococcus aureus, United States, 1999-2005. Emerging Infectious Diseases, 13(12), 1840-1846.
Li, T., Lu, H., Wang, X., Gao, Q., Dai, Y., Shang, J., & Li, M. (2017). Molecular characteristics of Staphylococcus aureus causing bovine mastitis between 2014 and 2015. Frontiers in Cellular and Infection Microbiology, 7, 127. https://doi.org/10.3389/fcimb.2017.00127
Luini, M., Cremonesi, P., Magro, G., Bianchini, V., Minozzi, G., Castiglioni, B., & Piccinini, R. (2015). Methicillin-resistant Staphylococcus aureus (MRSA) is associated with low within-herd prevalence of intramammary infections in dairy cows: Genotyping of isolates. Veterinary Microbiology, 178(3-4), 270-274.
Mehndiratta, P.L., & Bhalla, P. (2012). Typing of methicillin-resistant Staphylococcus aureus: A technical review. Indian Journal of Medical Research, 30(1), 16-23.
Resch, G., François, P., Morisset, D., Stojanov, M., Bonetti, E.J., Schrenzel, J., & Moreillon, P. (2013). Human-to-bovine jump of Staphylococcus aureus CC8 is associated with the loss of a β-hemolysin converting prophage and the acquisition of a new staphylococcal cassette chromosome. PLoS One, 8(3), e58187.
Ruppitsch, W., Indra, A., Stoger, A., Mayer, B., Stadlbauer, S., Wewalka, G., & Allerberger, F. (2006). Classifying spa types in complexes improves interpretation of typing results for methicillin-resistant Staphylococcus aureus. Journal of Clinical Microbiology, 44, 2442-2448.
Sakwinska, O., Giddey, M., Moreillon, M., Morisset, D., Waldvogel, A., & Moreillon, P. (2011). Staphylococcus aureus host range and human-bovine host shift. Applied and Environmental Microbiology, 77, 5908-5915.
Sharma, N., Rho, J.R., Hong, Y.H., Kang, T.Y., Lee, H.K., Hur, T.Y., & Jeong, D.K. (2012). Bovine mastitis: An Asiatic perspective. Asian Journal of Animal and Veterinary Advances, 7, 454-476.
Sheela, P., Isloor, R., Rathnamma, D., Veeregowda, B.M., Shambulingappa, B.E., Satyanarayana, M.L., Sundareshan, S., & Hegde, N.R. (2019). Characterisation by spa typing of Staphylococcus aureus isolates originating from bovine and bubaline mastitis in southern India. Indian Journal of Comparative Microbiology, Immunology and Infectious Diseases, 40(1), 1-10.
Silva, N.C.C., Guimarães, F.F., Manzi, M.P., Budri, P.E., Gómez-Sanz, E., Benito, D., & Torres, C. (2013). Molecular characterization and clonal diversity of methicillin-susceptible Staphylococcus aureus in milk of cows with mastitis in Brazil. Journal of Dairy Science, 96(11), 6856-6862.
Smith, E.M., Green, L.E., Medley, G.F., Bird, H.E., Fox, L.K., Schukken, Y.H., Kruze, J.V., Bradley, A.J., Zadoks, R.N., & Dowson, C.G. (2005). Multilocus sequence typing of intercontinental bovine Staphylococcus aureus isolates. Journal of Clinical Microbiology, 43, 4737-4743.
Sundareshan, S. (2012). Studies on phenotypic and molecular characterization of coagulase negative staphylococci isolated from clinical and subclinical cases of bovine mastitis (Ph.D. thesis). Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar, India.
Zadoks, R.N., Middleton, J.R., Mcdougall, S., Katholm, J., & Schukken, Y.H. (2011). Molecular epidemiology of mastitis pathogens of dairy cattle and comparative relevance to humans. Journal of Mammary Gland Biology & Neoplasia, 16(4), 357-372.
Downloads
Published
Issue
Section
License
Copyright (c) 2025 Indian Journal of Veterinary Sciences and Biotechnology

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.