11-Title: Antimicrobial resistance profiling of Staphylococcus aureus isolated from mastitic milk of bovine and dairy environment from arid and semi-arid regions of India
Authors: Taruna Bhati, Rajesh Chhabra, Rahul Yadav, Gaurav Charaya and Anil Kumar Kataria
Source: Ruminant Science (2021)-10(1):57-66.
How to cite this manuscript: Bhati Taruna, Chhabra Rajesh, Yadav Rahul, Charaya Gaurav and Kataria Anil Kumar (2021). Antimicrobial resistance profiling of Staphylococcus aureus isolated from mastitic milk of bovine and dairy environment from arid and semi-arid regions of India. Ruminant Science 10(1):57-66.
The present study was conducted on 197 samples comprising of 80 mastitic milk samples, 66 udder swabs and 51 swabs of milkers’ hands and were subjected to bacterial isolation, identification and determination of antibiogram pattern against 32 antimicrobials. One hundred and seven Staphylococcus aureus (S. aureus) isolates were isolated and were further characterized as methicillin-resistant S. aureus (MRSA)strains and for the presence of extended-spectrum of beta-lactamase (ESBL) activity. Isolates were molecularly identified as S. aureus by 23S rRNA gene-based ribotyping and nucgene amplification. Five antibiotics namely cefotaxime+clavulanic acid, imipenem, oxacillin, piperacillin+tazobactam, and vancomycin were found 100% effective against all the isolates followed by gentamicin (97.2%) and nitrofurantoin (96.3%). An overall higher resistance was shown towards penicillin (92.5%) and ampicillin (88.8%) but source-wise, mastitic milk isolates showed higher resistance towards ampicillin (90.2%) than penicillin (88.2%). There were 66 (61.7%) multi-drug resistant (MDR)isolates with MAR index ranging from 0.03 to 0.44. The majority of isolates from milkers’ hands (95.2%) were identified as MRSA by the MeReSa agar base method while the mecA gene was detected in 85.7%, 82.4% and 74.3% isolates from milker’s hands, udder and mastitic milk, respectively. Oxacillin E-strip method identified 21 (19.6%) MRSA isolates, all carrying the mecA gene. The detection of MDR strains and high prevalence of the mecA gene among S. aureus isolates is an alarming situation warranting careful monitoring of the resistance status of S. aureus in dairy environments as S. aureus transmission is dynamic and involves humans, animals as well as farm production environment.
Ateba C, Mbewe M, Moneoang MS and Bezuidenhout CC (2010). Antibiotic-resistant Staphylococcus aureus isolated from milk in the Mafikeng area, North West province, South Africa. South African Journal of Science 106:12-13.
Bauer AW, Kirby WM, Sherris JC and Turck M (1966). Antibiotic susceptibility testing by a standardized single disc method. American Journal of Clinical Pathology 45(4):493-496.
Beniæ M, Habrun B and Kompes G (2012). Clinical and epidemiological aspects of cow mastitis caused by Staphylococcus aureus and its methicillin-resistant strains. Medical Sciences Croatian Veterinary Institute, Zagreb, Croatia 37:113-122.
Bhati T, Gaurav K, Khichar V and Kataria AK (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.
Brakstad OG, Aasbakk K and Maeland JA (1992). Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. Journal of Clinical Microbiology 30(7):1654-60.
CLSI (2013). Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Third Informational Supplement (M100-S23). Clinical and Laboratory Standards Institute, Pennsylvania, USA. pp 72-81.
Madushanka DNN, Padmakumara HMS, Kumarasinghe GDN, Sanjeewa MPK and Magamage MPS (2017). Effect of two different bedding systems on udder health management of dairy cows. Ruminant Science 6(1):1-6.
Das Gunjan, Lalnunpuia C, Sarma K, Behera SK, Dutta TK and Bandyopadhyay Samiran (2015). Prevalence of Staphylococcus aureus associated sub-clinical mastitis in crossbred cows in Mizoram. Ruminant Science 4(2):167-170.
Diwakar, Akriti, Choudhary Sunita, Meena Dhirendra, Bhati Taruna and Kataria AK (2019). Antibiotic sensitivity pattern of some Staphylococcus aureus isolates from milk from goats with clinical mastitis. Ruminant Science 8(1):19-22.
Duncan DB (1955). Multiple Range and Multiple F-Test. Biometrics 11:1-5.
Hamid S, Bhat MA, Mir IA, Taku A, Badroo GA,Nazki S and Malik A (2017). Phenotypic and genotypic characterization of methicillin-resistant Staphylococcus aureus from bovine mastitis. Veterinary World 10(3):363-367.
Hanon BM (2017). Molecular Study of some virulence genes in biotype diversity of methicillin resistance Staphylococcus aureus isolated from handling carrier and bovine mastitis. International Journal of Science and Research 6 (2):1746-1752.
Hoque MN, Das ZC, Rahman ANMA, Haider MG and Islam MA (2018). Molecular characterization of Staphylococcus aureus strains in bovine mastitis milk in Bangladesh. International Journal of Veterinary Science and Medicine 6(1):53-60.
Ippolito G, Leone S, Lauria FN, Nicastri E and Wenzel RP (2010). Methicillin-resistant Staphylococcus aureus: the superbug. International Journal of Infectious Diseases 14(4):S7-S11.
Jamali H, Radmehr B and Ismail S (2014). Prevalence and antibiotic resistance of Staphylococcus aureus isolated from bovine clinical mastitis. Journal of Dairy Science 97(4):2226-2230.
Jamrozy DM, Fielder MD, Butaye P and Coldham NG (2012). Comparative genotypic and phenotypic characterisation of methicillin-resistant Staphylococcus aureus ST398 isolated from animals and humans. PLoS ONE 7(7):e40458.
Kalayu AA, Woldetsadik DA and Woldeamanuel Y (2020). Burden and antimicrobial resistance of S. aureus in dairy farms in Mekelle, Northern Ethiopia. BMC Veterinary Research 16:20.
Kumar Vaibhav, Patel JS, Patel BR, Mevada VK and Raval AP (2012). Therapeutic efficacy of antimicrobial drugs in clinical mastitis of cross bred cattle. Ruminant Science 1(2):177-180.
Krumperman PH (1983). Multiple antibiotic resistance indexing of Escherichia coli to identify high-risk sources of fecal contamination of foods. Applied and Environmental Microbiology 46:165-170.
Livermore DM and Brown DFJ (2001). Detection of ß-lactamase mediated resistance. Journal of Antimicrobial Chemotherapy 48:59-64.
Marques VF, da Motta CC, Soares BdaS, Melo DA, de Coelho, SdeMdeO, Coelho I da S and Souza MMSde (2017). Biofilm production and beta-lactamic resistance in Brazilian Staphylococcus aureus isolates from bovine mastitis. Brazilian Journal of Microbiology 48(1):118-124.
Mehrotra M, Wang G and Johnson WM (2000). Multiplex PCR for detection of genes for Staphylococcus aureus enterotoxins, exfoliative toxins, toxic shock syndrome toxin 1, and methicillin resistance. Journal of Clinical Microbiology 38(3):1032-1035.
Murakami K,Minamide W, Wada K, Nakamura E and Teraoka H (1991). Identification of methicillin-resistant strains of staphylococci by polymerase chain reaction. Journal of Clinical Microbiology 29:2240-2244.
Nirmali WKR, Priyabhashana AHL, Bandara AMS and Magamage MPS (2018). Assessment of milk quality of upcountry diary farm in Sri Lanka. Ruminant Science 7(1):1-4.
NCCLS (1992). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. 2nd Edn, Approved Standard. NCCLS Document M7-A2 (M100-S4).
Pantosti A, Sanchini A and Monaco M (2007). Mechanisms of antibiotic resistance in S. aureus. Future Microbiology 2:323-334.
Parth FM, Chauhan HC, Bhagat AG, Chandel BS, Patel MV, Dadawala AI and Kher HN (2016). Detection of virulence associated factors from Staphylococcus aureus isolated from bovine mastitis. Buffalo Bulletin 35(4):687-696.
Pereira V, Lopes C, Castro A, Silva J, Gibbs P and Teixeira P (2009).Characterization for enterotoxin production, virulence factors, and antibiotic susceptibility of S. aureus isolated from various foods in Portugal. Food Microbiology 26(3):278-282.
Quinn PJ, Carter ME, Markey BK and Carter GR (1994). Clinical Veterinary Microbiology. Mosby-Year Book Europe Ltd, England.
Raguvaran R, Mondal DB, Jithin MV, Kumar Bipin and Sivakumar M (2016). Staphylococcus aureus mastitis in a nondescript goat. Ruminant Science 5(2):291-292.
Raorane Abhay, Chothe Shubhadha, Dubal ZB, Barbuddhe SB, Karunakaran M, Doijad Swapnil, Pathak Ajay, Poharkar Krupali and Singh NP (2013). Antimicrobial resistance of the pathogens isolated from bovine mastitis in Goa. Ruminant Science 2(2):139-144.
Ren Q, Liao G, Wu Z, Lv J and Chen W (2020). Prevalence and characterization of Staphylococcus aureus isolates from subclinical bovine mastitis in southern Xinjiang, China. Journal of Dairy Science 103:3368-3380.
Rohlf FJ (1998). NTSYSpc Numerical taxonomy and multivariate analysis system version 2.0 user guide. Applied Biostatistics Inc., Setauket, New York. p 37.
Sanjiv K and Kataria AK (2006). Antibiogram of Staphylococcus aureus isolates of cattle clinical mastitis origin. Veterinary Practitioner 7 (2):123-125.
Savita, Singh AP, Nayak TC, Chahar A, Yadav R and Kachhawa JP (2020). Isolation and identification of bacteria in subclinical mastitis in cattle from Bikaner city. Ruminant Science 9(1):45-48.
Shabaz SS, Prameela DR, Sreenivasulu D and Sujatha K (2020). Evaluation of on-farm milk culture system for identification of mastitis pathogens. Ruminant Science 9(2):363-372.
Shamila-Syuhada AK, Rusul G, Wan-Nadiah WA and Chuah LO (2016). Prevalence and antibiotics resistance of Staphylococcus aureus isolates isolated from raw milk obtained from small-scale dairy farms in Penang, Malaysia. Pakistan Veterinary Journal 36(1):98-102.
Singh Pawanjit, Nigam Rajesh, Kumar Amit and Pandey Vijay (2018a). Isolation and molecular characterization of pathogens associated with mastitis in Sahiwal cows. Ruminant Science 7(1):43-46.
Singh A, Joshi RK, Joshi N and Singh P (2018b). Isolation and identification of multidrug resistant and methicillin resistant Staphylococcus aureus from bovine. International Journal of Current Microbiology and Applied Sciences 7:230-238.
Straub JA, Hertel C and Hammes WP (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):1150-1156.
Sunita, Diwakar and Kataria AK (2017). Antibiotic resistance pattern of Staphylococcus aureus isolated from milk of cattle with clinical mastitis. Ruminant Science 6(2):319-322.
Suleiman AB, Kwaga JKP, Umoh VJ, Okolocha EC, Muhammed M, Lammler C, Shaibu SJ, Akineden O and Weiss R (2012). Macro-restriction analysis of Staphylococcus aureus isolated from subclinical bovine mastitis in Nigeria. African Journal of Microbiology Research 6(33):6270-6274.
Xu J, Tan X, Zhang X, Xia X and Sun H (2015). The diversities of staphylococcal species, virulence and antibiotic resistance genes in the subclinical mastitis milk from a single Chinese cow herd. Microbial Pathogens 88:29.