Characterization of Staphylococcus aureus isolates from mastitic milk, udder surfaces and milkers' hands from arid and semi arid regions of India for capsular (cap5K and cap8K) and collagen adhesin (cna) genes
313 / 149
Keywords:
Capsular types, cna gene, Duplex PCR, Mastitis, Staphylococcus aureusAbstract
Staphylococcus aureus (S. aureus) is an important cause of bovine mastitis. The antiphagocytic capsule of bacteria promotes colonization and persistence on mucosal surfaces while the presence of collagen adhesin (cna gene) is associated with bacterial adhesion in mammary tissues; hence both are regarded as important determinants of virulence. In the present study, 197 samples consisting of mastitic milk, milkers' hand swabs and udder surface swabs were collected which yielded 107 S. aureus isolates. These S. aureus isolates (107) from different sources of sampling were characterized for their capsular types targeting cap5K and cap8K genes in a duplex PCR along with amplification of cna gene. Majority of the isolates (67.6%) possessed cap5K gene with a lower percentage (22.9%) of isolates carrying cap8K gene and 9.5% carrying both the genes. Moreover, cap5K gene was predominant in isolates from milkers’ hands (85.7%) while cap8K gene was more common in bovine udder isolates (41.2%). cna gene was observed in 27.6% isolates with highest occurrence in milk isolates (44.8%) compared to udder (37.9%) and milkers' hand (17.2%). cna positive isolates carried cap8K gene (66.7%) and were significantly associated with both cap5k and cap8k genes. In conclusion, the majority of the S. aureus isolates of mastitis obtained from arid and semi arid zones of India possessed the genes for capsule production, and cap5K was the predominant gene. The cna gene for collagen adhesion was observed in fewer isolates with significant association with cap8K gene.
Downloads
References
Ahangari Z, Ghorbanpoor M, Shapouri M R S, Gharibi D and Ghazvini K. 2017. Methicillin resistance and selective genetic determinants of Staphylococcus aureus isolates with bovine mastitis milk origin. Iranian Journal of Microbiology 9(3): 152–59
Aslantaº O and Demir C. 2016. Investigation of the antibiotic resistance and biofilm forming ability of Staphylococcus aureus from subclinical bovine mastitis cases. Journal of Dairy Science 99(11): 8607–13. DOI: https://doi.org/10.3168/jds.2016-11310
Bhati T, Gaurav K, Khichar V and Kataria A K. 2018. Prevalence of Staphylococcus aureus isolated from mastitic milk, udder surfaces and milkers’ hands from different farms in Bikaner, Rajasthan. Journal of Animal Research 8(5): 867–72. DOI: https://doi.org/10.30954/2277-940X.10.2018.19
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(1): 345–52 DOI: https://doi.org/10.1128/IAI.69.1.345-352.2001
Brakstad O G, Aasbakk K and Maeland J A. 1992. Detection of Staphylococcus aureus by polymerase chain reaction amplification of the gene. Journal of Clinical Microbiology 30(7): 1654–60. DOI: https://doi.org/10.1128/jcm.30.7.1654-1660.1992
Camuss one C, Rejf P, Pujato N, Schwab A, Marcipar I and Calvinho L F. 2012. Genotypic and phenotypic detection of capsular polysaccharides in Staphylococcus aureus isolated from bovine intramammary infections in Argentina. Brazilian Journal of Microbiology 43 1010–14 DOI: https://doi.org/10.1590/S1517-83822012000300023
El-Sayed A, Alber J, Lammler C, Jager S, Wolter W and Castaneda-Vazquez H. 2006. Comparative study on genotypic properties of Staphylococcus aureus isolated from clinical and subclinical mastitis in Mexico. Veterinaria México 37(2): 165– 79
Haveri M, Roslöf A, Rantala L and Pyörälä S. 2007. Virulence genes of bovine Staphylococcus aureus from persistent and nonpersistent intramammary infections with different clinical characteristics. Journal of Applied Microbiology 103 993– 1000 DOI: https://doi.org/10.1111/j.1365-2672.2007.03356.x
Ikawaty R, Brouwer E C, Duijkeren V E, Mevius D, Verhoef J and Fluet A C. 2010. Virulence factors of genotyped bovine mastitis Staphylococcus aureus isolates in the Netherlands International Journal of Dairy Science 5(2): 60–70. DOI: https://doi.org/10.3923/ijds.2010.60.70
Khichar V and Kataria A K. 2014. Capsular genotyping (cap5k and cap8k) of Staphylococcus aureus isolates from cattle with clinical mastitis. HVM International Journal of the Bioflux Society 6(1): 30–33.
Krithiga N, Antony P X, Tollersrud T, Mukhopadhyay H K, Pillai R M, Vijayalakshmi P and Thanislass J. 2018. Molecular typing of capsular polysaccharides of Staphylococcus aureus isolated from cases of bovine mastitis by PCR. Concepts of Dairy and Veterinary Science (1): 13–16. DOI: https://doi.org/10.32474/CDVS.2018.01.000102
Kumar R, Yadav B R and Singh R S. 2011. Antibiotic resistance and pathogenicity factors in Staphylococcus aureus isolated from mastitic Sahiwal cattle. Journal of Biosciences 36(1): 175–88 DOI: https://doi.org/10.1007/s12038-011-9004-6
van Leeuwen W, Melles D C, Alaidan A, Al-Ahdal M, Boelens H A M, Snijders S V, Wertheim H, van Duijkeren E et al 2005. Host- and tissue-specific pathogenic traits of Staphylococcus aureus Journal of Bacteriology 187 4584– 91. DOI: https://doi.org/10.1128/JB.187.13.4584-4591.2005
Markey B K, Leonard F, Archambault M, Culinane A and Maguire D. 2013. Clinical Veterinary Microbiology. 2nd Edn. 90 pages. Mosby/Elsevier, Edinburgh, Scotland.
Melchior M B, van Osch M H, Graat R M, van Duijkeren E, Mevius D J, Nielen M, Gaastra W and Fink-Gremmels J. 2009. Biofilm formation and genotyping of Staphylococcus aureus bovine mastitis isolates: evidence for lack of penicillin resistance in Agr-type II strains. Journal of Veterinary Microbiology 28: 83–89. DOI: https://doi.org/10.1016/j.vetmic.2008.12.004
Memon J, Yang Y, Kashif J, Yaqoob M, Buriro R, Soomro J, Liping W and Hongjie F. 2013. Genotypes, virulence factors and antimicrobial resistance genes of Staphylococcus aureus isolated in bovine subclinical mastitis from Eastern China Pakistan Veterinary Journal 33(4): 486–91.
Nashev D, Toshkova K, Salasia S I, Hassan AA. Lammler C and Zschock M. 2004. Distribution of virulence genes of Staphylococcus aureus isolated from stable nasal carriers FEMS Microbiology Letters 233 45–52 DOI: https://doi.org/10.1016/j.femsle.2004.01.032
Nathawat P, Bhati T, Sharma S K, Yadav R and Kataria A K. 2015. Characterization of Staphylococcus aureus of goat mastitis milk origin for cap and clfA genes. Journal of Pure and Applied Microbiology 9(2): 1055–61.
O’Riordan K and Lee J C. 2004 Staphylococcus aureus capsular polysaccharides. Clinical Microbiology Reviews 17(1): 218–234 DOI: https://doi.org/10.1128/CMR.17.1.218-234.2004
Pereyra E A, Picech F, Renna M S, Baravalle C, Andreotti C S, Russi R, Calvinho L F, Diez C, Dallard B E. 2016. Detection of Staphylococcus aureus adhesion and biofilm-producing genes and their expression during internalization in bovine mammary epithelial cells. Veterinary Microbiology 183: 69– 77 DOI: https://doi.org/10.1016/j.vetmic.2015.12.002
Proietti P C, Coppola G, Bietta A, Marenzoni M L, Hyatt D R, Coletti M and Passamonti F. 2010. Characterization of genes encoding virulence determinants and toxins in Staphylococcus r from bovine milk in Central Italy. Journal of Veterinary and Medical Science 72(11): 1443–48. DOI: https://doi.org/10.1292/jvms.10-0158
Reinoso E B, El-Sayed A, Lämmler C, Bogni C and Zschöck M. 2008. Genotyping of Staphylococcus aureus isolated from humans, bovine subclinical mastitis and food samples in Argentina. Microbiology Research 163 (3): 314–22. DOI: https://doi.org/10.1016/j.micres.2006.05.013
Ryding U, Flock J I, Flock M, Soderquist B and Cristensson B. 1997. Expression of collagen-binding protein and types 5 and 8 capsular polysaccharide in clinical isolates of Staphylococcus aureus Journal of Infectious Diseases 176: 1096–1009. DOI: https://doi.org/10.1086/516520
Saei H D. 2012. Distribution of collagen adhesin gene among various types of Staphylococcus aureus strains associated with bovine mammary gland. Comparative Clinical Pathology 21 571–76 DOI: https://doi.org/10.1007/s00580-010-1136-9
Salasia S I O, Khusnan Z, Lammler C and Zschock M. 2004. Comparative studies on phenotypic 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(2): 103–09. DOI: https://doi.org/10.4142/jvs.2004.5.2.103
Salimena A P, Lange C C, Camussone C, Signorini M, Calvinho L F, Brito M A, Borges C A, Guimarães A S et al 2016 Genotypic and phenotypic detection of capsular polysaccharide and biofilm formation in Staphylococcus r isolated from bovine milk collected from Brazilian dairy farms. Veterinary Research Communications 40(3–4): 97–106 DOI: https://doi.org/10.1007/s11259-016-9658-5
Schmidt T, Kock M M and Ehlers M M. 2017. Molecular characterization of Staphylococcus aureus isolated from bovine mastitis and close human contacts in South African dairy herds: Genetic diversity and inter-species host transmission. Frontiers in Microbiology 8 511 DOI: https://doi.org/10.3389/fmicb.2017.00511
Sharma S K, Mehta S C and Kataria A K. 2016. Capsular typing of Staphylococcus r isolates from camel and other domestic animals using duplex polymerase chain reaction. Journal of Camel Practice and Research 23(1): 81–84. DOI: https://doi.org/10.5958/2277-8934.2016.00011.4
Soares B S, Melo D A, Motta C C, Marques V F, Barreto N B, Coelho S M O, Coelho I S and Souza M M S. 2017. Characterization of virulence and antibiotic profile and agrtyping of Staphylococcus aureus from milk of subclinical mastitis bovine in State of Rio de Janeiro. Arquivo Brasileiro de Medicina Veterinária e Zootecnia 69(4): 843–50. DOI: https://doi.org/10.1590/1678-4162-9260
Straub J A, Hertel C and Hammes W P. 1999. A 23S rRNA target polymerase chain reaction based system for detection of Staphylococcus aureus in meat starter cultures and dairy products. Journal of Food Protection 62(10): 1150–56. DOI: https://doi.org/10.4315/0362-028X-62.10.1150
Tollersrud T, Kenny K, Reitz A J and Lee J C. 2000. Genetic and serologic evaluation of capsule production by bovine mammary isolates of Staphylococcus aureus and other Staphylococcus spp. from Europe and the United States. Journal of Clinical Microbiology 38(8): 2998–3003. DOI: https://doi.org/10.1128/JCM.38.8.2998-3003.2000
Upadhyay A, Kataria A K, Sharma R and Singh G. 2010. Capsular typing of Staphylococcus aureus isolates from cattle and goat mastitis by PCR targeting cap5K and cap8K genes. Indian Journal of Animal Sciences 80(11): 1062–65.
Veh K A, Klein R C, Ster C, Keefe G, Lacasse P, Scholl D, Roy J P, Haine D, Dufour S, Talbot B G, Ribon A O and Malouin F. 2015. Genotypic and phenotypic characterization of Staphylococcus aureus causing persistent and nonpersistent subclinical bovine intramammary infections during lactation or the dry period. Journal of Dairy Science 98 155–68 DOI: https://doi.org/10.3168/jds.2014-8044
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(3): 725–29. DOI: https://doi.org/10.1128/JCM.01572-06
Yadav R, Sharma S K, Yadav J, Nathawat P and Kataria A K. 2015. Phenotypic and genotypic characterization of Staphylococcus aureus of mastitic milk origin from cattle and buffalo for some virulence properties. Journal of Pure and Applied Microbiology 9(1): 425–31.
Zecconi A, Cesaris L, Liandris E, Daprà V and Piccinini R. 2006. Role of several Staphylococcus aureus virulence factors on the inflammatory response in bovine mammary gland. Microbial Pathogenesis 40(4): 177–83. DOI: https://doi.org/10.1016/j.micpath.2006.01.001
Downloads
Submitted
Published
Issue
Section
License
Copyright (c) 2019 The Indian Journal of Animal Sciences

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
The copyright of the articles published in The Indian Journal of Animal Sciences is vested with the Indian Council of Agricultural Research, which reserves the right to enter into any agreement with any organization in India or abroad, for reprography, photocopying, storage and dissemination of information. The Council has no objection to using the material, provided the information is not being utilized for commercial purposes and wherever the information is being used, proper credit is given to ICAR.