Alarming multidrug resistance in Staphylococcus aureus isolated from raw milk of cows with subclinical mastitis: Antibiotic resistance patterns and occurrence of selected resistance genes
Bovine mastitis is a widespread and costly disease that affects dairy farming globally, characterized by mammary gland inflammation. Bovine intramammary gland infection has been associated with more than 135 different pathogens of which Staphylococcus aureus is the main etiology of sub-clinical mast...
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| description | Bovine mastitis is a widespread and costly disease that affects dairy farming globally, characterized by mammary gland inflammation. Bovine intramammary gland infection has been associated with more than 135 different pathogens of which Staphylococcus aureus is the main etiology of sub-clinical mastitis (SCM). The current study was designed to investigate the prevalence, antibiotic resistance pattern, and the presence of antibiotic resistance genes (mecA, tetK, aacA-aphD and blaZ) in S. aureus isolated from the raw milk of cows with subclinical mastitis. A total of 543 milk samples were collected from lactating cows such as Holstein Friesian (n = 79), Sahiwal (n = 175), Cholistani (n = 107), and Red Sindhi (n = 182) from different dairy farms in Pakistan. From the milk samples microscopic slides were prepared and the somatic cell count was assessed to find SCM. To isolate and identify S. aureus, milk was streaked on mannitol salt agar (MSA) plates. Further confirmation was done based on biochemical assays, including gram staining (+ coccus), catalase test (+), and coagulase test (+). All the biochemically confirmed S. aureus isolates were molecularly identified using the thermonuclease (nuc) gene. The antibiotic resistance pattern of all the S. aureus isolates was evaluated through the disc diffusion method. Out of 543 milk samples, 310 (57.09%) were positive for SCM. Among the SCM-positive samples, S. aureus was detected in 30.32% (94/310) samples. Out of 94 isolates, 47 (50%) were determined to be multidrug resistant (MDR). Among these MDR isolates, 11 exhibited resistance to Cefoxitin, and hence were classified as methicillin-resistant Staphylococcus aureus (MRSA). The S. aureus isolates showed the highest resistance to Lincomycin (84.04%) followed by Ampicillin (45.74%), while the least resistance was shown to Sulfamethoxazole/Trimethoprim (3.19%) and Gentamycin (6.38%). Polymerase chain reaction (PCR) analysis revealed that 55.31% of the isolates carried blaZ gene, 46.80% carried tetK gene, 17.02% harbored the mecA gene, whereas, aacA-aphD gene was found in 13.82% samples. Our findings revealed a significant level of contamination of milk with S. aureus and half (50%) of the isolates were MDR. The isolated S. aureus harbored various antibiotic resistance genes responsible for the absorbed phenotypic resistance. The alarmingly high prevalence of MDR S. aureus isolates and MRSA strains in these cases possess a serious risk to public health, emphasizes t |
| doi_str_mv | 10.1371/journal.pone.0301200 |
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Bovine intramammary gland infection has been associated with more than 135 different pathogens of which Staphylococcus aureus is the main etiology of sub-clinical mastitis (SCM). The current study was designed to investigate the prevalence, antibiotic resistance pattern, and the presence of antibiotic resistance genes (mecA, tetK, aacA-aphD and blaZ) in S. aureus isolated from the raw milk of cows with subclinical mastitis. A total of 543 milk samples were collected from lactating cows such as Holstein Friesian (n = 79), Sahiwal (n = 175), Cholistani (n = 107), and Red Sindhi (n = 182) from different dairy farms in Pakistan. From the milk samples microscopic slides were prepared and the somatic cell count was assessed to find SCM. To isolate and identify S. aureus, milk was streaked on mannitol salt agar (MSA) plates. Further confirmation was done based on biochemical assays, including gram staining (+ coccus), catalase test (+), and coagulase test (+). All the biochemically confirmed S. aureus isolates were molecularly identified using the thermonuclease (nuc) gene. The antibiotic resistance pattern of all the S. aureus isolates was evaluated through the disc diffusion method. Out of 543 milk samples, 310 (57.09%) were positive for SCM. Among the SCM-positive samples, S. aureus was detected in 30.32% (94/310) samples. Out of 94 isolates, 47 (50%) were determined to be multidrug resistant (MDR). Among these MDR isolates, 11 exhibited resistance to Cefoxitin, and hence were classified as methicillin-resistant Staphylococcus aureus (MRSA). The S. aureus isolates showed the highest resistance to Lincomycin (84.04%) followed by Ampicillin (45.74%), while the least resistance was shown to Sulfamethoxazole/Trimethoprim (3.19%) and Gentamycin (6.38%). Polymerase chain reaction (PCR) analysis revealed that 55.31% of the isolates carried blaZ gene, 46.80% carried tetK gene, 17.02% harbored the mecA gene, whereas, aacA-aphD gene was found in 13.82% samples. Our findings revealed a significant level of contamination of milk with S. aureus and half (50%) of the isolates were MDR. The isolated S. aureus harbored various antibiotic resistance genes responsible for the absorbed phenotypic resistance. The alarmingly high prevalence of MDR S. aureus isolates and MRSA strains in these cases possess a serious risk to public health, emphasizes the urgent need to address this issue to protect both human and animal health in Pakistan.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0301200</identifier><identifier>PMID: 38753608</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Ampicillin ; Analysis ; Animal health ; Animals ; Anti-Bacterial Agents - pharmacology ; Antibiotic resistance ; Antibiotics ; Bacteria ; Bacterial Proteins - genetics ; Biology and Life Sciences ; Catalase ; Catalase test ; Cattle ; Cefoxitin ; Coagulase ; Coagulase test ; Cow's milk ; Dairy farming ; Dairy farms ; Development and progression ; Drug resistance ; Drug resistance in microorganisms ; Drug Resistance, Multiple, Bacterial - genetics ; Female ; Food contamination ; Genes ; Genetic research ; Gentamicin ; Immune system ; Infection ; Lincomycin ; Mammary gland ; Mammary glands ; Mannitol ; Mastitis ; Mastitis, Bovine - epidemiology ; Mastitis, Bovine - microbiology ; MecA protein ; Medicine and Health Sciences ; Methicillin ; Microbial Sensitivity Tests ; Milk ; Milk - microbiology ; Milk production ; Multidrug resistance ; Nosocomial infections ; Pakistan - epidemiology ; Pathogens ; Plasmids ; Polymerase chain reaction ; Proteins ; Public health ; Ribonucleic acid ; RNA ; Skin ; Staphylococcal infections ; Staphylococcal Infections - epidemiology ; Staphylococcal Infections - microbiology ; Staphylococcal Infections - veterinary ; Staphylococcus aureus ; Staphylococcus aureus - drug effects ; Staphylococcus aureus - genetics ; Staphylococcus aureus - isolation & purification ; Staphylococcus infections ; Sulfamethoxazole ; Tetracycline ; Tetracyclines ; Thermonuclease ; Toxins ; Trimethoprim ; Virulence ; Wildlife conservation</subject><ispartof>PloS one, 2024-05, Vol.19 (5), p.e0301200-e0301200</ispartof><rights>Copyright: © 2024 Haq et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</rights><rights>COPYRIGHT 2024 Public Library of Science</rights><rights>2024 Haq et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2024 Haq et al 2024 Haq et al</rights><rights>2024 Haq et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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Bovine intramammary gland infection has been associated with more than 135 different pathogens of which Staphylococcus aureus is the main etiology of sub-clinical mastitis (SCM). The current study was designed to investigate the prevalence, antibiotic resistance pattern, and the presence of antibiotic resistance genes (mecA, tetK, aacA-aphD and blaZ) in S. aureus isolated from the raw milk of cows with subclinical mastitis. A total of 543 milk samples were collected from lactating cows such as Holstein Friesian (n = 79), Sahiwal (n = 175), Cholistani (n = 107), and Red Sindhi (n = 182) from different dairy farms in Pakistan. From the milk samples microscopic slides were prepared and the somatic cell count was assessed to find SCM. To isolate and identify S. aureus, milk was streaked on mannitol salt agar (MSA) plates. Further confirmation was done based on biochemical assays, including gram staining (+ coccus), catalase test (+), and coagulase test (+). All the biochemically confirmed S. aureus isolates were molecularly identified using the thermonuclease (nuc) gene. The antibiotic resistance pattern of all the S. aureus isolates was evaluated through the disc diffusion method. Out of 543 milk samples, 310 (57.09%) were positive for SCM. Among the SCM-positive samples, S. aureus was detected in 30.32% (94/310) samples. Out of 94 isolates, 47 (50%) were determined to be multidrug resistant (MDR). Among these MDR isolates, 11 exhibited resistance to Cefoxitin, and hence were classified as methicillin-resistant Staphylococcus aureus (MRSA). The S. aureus isolates showed the highest resistance to Lincomycin (84.04%) followed by Ampicillin (45.74%), while the least resistance was shown to Sulfamethoxazole/Trimethoprim (3.19%) and Gentamycin (6.38%). Polymerase chain reaction (PCR) analysis revealed that 55.31% of the isolates carried blaZ gene, 46.80% carried tetK gene, 17.02% harbored the mecA gene, whereas, aacA-aphD gene was found in 13.82% samples. Our findings revealed a significant level of contamination of milk with S. aureus and half (50%) of the isolates were MDR. The isolated S. aureus harbored various antibiotic resistance genes responsible for the absorbed phenotypic resistance. The alarmingly high prevalence of MDR S. aureus isolates and MRSA strains in these cases possess a serious risk to public health, emphasizes the urgent need to address this issue to protect both human and animal health in Pakistan.</description><subject>Ampicillin</subject><subject>Analysis</subject><subject>Animal health</subject><subject>Animals</subject><subject>Anti-Bacterial Agents - pharmacology</subject><subject>Antibiotic resistance</subject><subject>Antibiotics</subject><subject>Bacteria</subject><subject>Bacterial Proteins - genetics</subject><subject>Biology and Life Sciences</subject><subject>Catalase</subject><subject>Catalase test</subject><subject>Cattle</subject><subject>Cefoxitin</subject><subject>Coagulase</subject><subject>Coagulase test</subject><subject>Cow's milk</subject><subject>Dairy farming</subject><subject>Dairy farms</subject><subject>Development and progression</subject><subject>Drug resistance</subject><subject>Drug resistance in microorganisms</subject><subject>Drug Resistance, Multiple, Bacterial - genetics</subject><subject>Female</subject><subject>Food contamination</subject><subject>Genes</subject><subject>Genetic research</subject><subject>Gentamicin</subject><subject>Immune system</subject><subject>Infection</subject><subject>Lincomycin</subject><subject>Mammary gland</subject><subject>Mammary glands</subject><subject>Mannitol</subject><subject>Mastitis</subject><subject>Mastitis, Bovine - epidemiology</subject><subject>Mastitis, Bovine - microbiology</subject><subject>MecA protein</subject><subject>Medicine and Health Sciences</subject><subject>Methicillin</subject><subject>Microbial Sensitivity Tests</subject><subject>Milk</subject><subject>Milk - microbiology</subject><subject>Milk production</subject><subject>Multidrug resistance</subject><subject>Nosocomial infections</subject><subject>Pakistan - epidemiology</subject><subject>Pathogens</subject><subject>Plasmids</subject><subject>Polymerase chain reaction</subject><subject>Proteins</subject><subject>Public health</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>Skin</subject><subject>Staphylococcal infections</subject><subject>Staphylococcal Infections - epidemiology</subject><subject>Staphylococcal Infections - microbiology</subject><subject>Staphylococcal Infections - veterinary</subject><subject>Staphylococcus aureus</subject><subject>Staphylococcus aureus - drug effects</subject><subject>Staphylococcus aureus - genetics</subject><subject>Staphylococcus aureus - isolation & purification</subject><subject>Staphylococcus infections</subject><subject>Sulfamethoxazole</subject><subject>Tetracycline</subject><subject>Tetracyclines</subject><subject>Thermonuclease</subject><subject>Toxins</subject><subject>Trimethoprim</subject><subject>Virulence</subject><subject>Wildlife conservation</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk91u1DAQhSMEoqXwBggsISG42MWOEzvmBq0qfipVqkSBW8txJlkXx15sh9Jn4-Vw6LbaRb0gkeJo8s05nomnKJ4SvCSUkzcXfgpO2eXGO1hiikmJ8b3ikAhaLliJ6f2d94PiUYwXGNe0YexhcUAbXlOGm8Pi98qqMBo3oHGyyXRhGlCAaGJSTgMyDp0ntVlfWa-91lNEagqQFxO9VQk61Ac_oqAu0Wjsd-R7pP1lRJcmrVGcWm2NM1pZNKqYTDLxLVq5ZFrjk9G7RhuVEgSX9V2HZqcQYI5nwQgW9Gy1gw_gID4uHvTKRniyXY-Krx_efzn-tDg9-3hyvDpdaFaVadETWpNS0IrlJyOYMQBcM9VAw6u67RRXineY1bziNdai77q2b3EldC8YFYoeFc-vdTfWR7nte5QUM1E2XJQ0EyfXROfVhdwEM6pwJb0y8m_Ah0GqkCu2IIHqmjPe4FKIqmO8JVxoQbu-7aAqG5K13m3dpnaEToNLQdk90f0vzqzl4H9KQrBo8qaywqutQvA_JohJjiZqsFY58NO88Zrli4g6oy_-Qe8ub0sNKldgXO-zsZ5F5YqLKncU8ypTyzuofHcwGp1PaW9yfC_h9V5CZhL8SoOaYpQn55__nz37ts--3GHXoGxa5-M6JeNd3Aera1AHH2OA_rbLBMt5yG66Iechk9shy2nPdv_QbdLNVNE_SEMmFg</recordid><startdate>20240516</startdate><enddate>20240516</enddate><creator>Haq, Ijaz Ul</creator><creator>Kamal, Mustafa</creator><creator>Swelum, Ayman A</creator><creator>Khan, Shehryar</creator><creator>Ríos-Escalante, Patricio R De Los</creator><creator>Usman, Tahir</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0002-8192-5800</orcidid><orcidid>https://orcid.org/0000-0001-5729-8037</orcidid></search><sort><creationdate>20240516</creationdate><title>Alarming multidrug resistance in Staphylococcus aureus isolated from raw milk of cows with subclinical mastitis: Antibiotic resistance patterns and occurrence of selected resistance genes</title><author>Haq, Ijaz Ul ; Kamal, Mustafa ; Swelum, Ayman A ; Khan, Shehryar ; Ríos-Escalante, Patricio R De Los ; Usman, Tahir</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c642t-f13512934612961066ee056a8e8745bda7aa7d06574750c9fddbfb049cf9639a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>Ampicillin</topic><topic>Analysis</topic><topic>Animal health</topic><topic>Animals</topic><topic>Anti-Bacterial Agents - pharmacology</topic><topic>Antibiotic resistance</topic><topic>Antibiotics</topic><topic>Bacteria</topic><topic>Bacterial Proteins - genetics</topic><topic>Biology and Life Sciences</topic><topic>Catalase</topic><topic>Catalase test</topic><topic>Cattle</topic><topic>Cefoxitin</topic><topic>Coagulase</topic><topic>Coagulase test</topic><topic>Cow's milk</topic><topic>Dairy farming</topic><topic>Dairy farms</topic><topic>Development and progression</topic><topic>Drug resistance</topic><topic>Drug resistance in microorganisms</topic><topic>Drug Resistance, Multiple, Bacterial - genetics</topic><topic>Female</topic><topic>Food contamination</topic><topic>Genes</topic><topic>Genetic research</topic><topic>Gentamicin</topic><topic>Immune system</topic><topic>Infection</topic><topic>Lincomycin</topic><topic>Mammary gland</topic><topic>Mammary glands</topic><topic>Mannitol</topic><topic>Mastitis</topic><topic>Mastitis, Bovine - epidemiology</topic><topic>Mastitis, Bovine - microbiology</topic><topic>MecA protein</topic><topic>Medicine and Health Sciences</topic><topic>Methicillin</topic><topic>Microbial Sensitivity Tests</topic><topic>Milk</topic><topic>Milk - microbiology</topic><topic>Milk production</topic><topic>Multidrug resistance</topic><topic>Nosocomial infections</topic><topic>Pakistan - epidemiology</topic><topic>Pathogens</topic><topic>Plasmids</topic><topic>Polymerase chain reaction</topic><topic>Proteins</topic><topic>Public health</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>Skin</topic><topic>Staphylococcal infections</topic><topic>Staphylococcal Infections - epidemiology</topic><topic>Staphylococcal Infections - microbiology</topic><topic>Staphylococcal Infections - veterinary</topic><topic>Staphylococcus aureus</topic><topic>Staphylococcus aureus - drug effects</topic><topic>Staphylococcus aureus - genetics</topic><topic>Staphylococcus aureus - isolation & purification</topic><topic>Staphylococcus infections</topic><topic>Sulfamethoxazole</topic><topic>Tetracycline</topic><topic>Tetracyclines</topic><topic>Thermonuclease</topic><topic>Toxins</topic><topic>Trimethoprim</topic><topic>Virulence</topic><topic>Wildlife conservation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Haq, Ijaz Ul</creatorcontrib><creatorcontrib>Kamal, Mustafa</creatorcontrib><creatorcontrib>Swelum, Ayman A</creatorcontrib><creatorcontrib>Khan, Shehryar</creatorcontrib><creatorcontrib>Ríos-Escalante, Patricio R De Los</creatorcontrib><creatorcontrib>Usman, Tahir</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - Academic</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials Science Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Haq, Ijaz Ul</au><au>Kamal, Mustafa</au><au>Swelum, Ayman A</au><au>Khan, Shehryar</au><au>Ríos-Escalante, Patricio R De Los</au><au>Usman, Tahir</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Alarming multidrug resistance in Staphylococcus aureus isolated from raw milk of cows with subclinical mastitis: Antibiotic resistance patterns and occurrence of selected resistance genes</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2024-05-16</date><risdate>2024</risdate><volume>19</volume><issue>5</issue><spage>e0301200</spage><epage>e0301200</epage><pages>e0301200-e0301200</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Bovine mastitis is a widespread and costly disease that affects dairy farming globally, characterized by mammary gland inflammation. Bovine intramammary gland infection has been associated with more than 135 different pathogens of which Staphylococcus aureus is the main etiology of sub-clinical mastitis (SCM). The current study was designed to investigate the prevalence, antibiotic resistance pattern, and the presence of antibiotic resistance genes (mecA, tetK, aacA-aphD and blaZ) in S. aureus isolated from the raw milk of cows with subclinical mastitis. A total of 543 milk samples were collected from lactating cows such as Holstein Friesian (n = 79), Sahiwal (n = 175), Cholistani (n = 107), and Red Sindhi (n = 182) from different dairy farms in Pakistan. From the milk samples microscopic slides were prepared and the somatic cell count was assessed to find SCM. To isolate and identify S. aureus, milk was streaked on mannitol salt agar (MSA) plates. Further confirmation was done based on biochemical assays, including gram staining (+ coccus), catalase test (+), and coagulase test (+). All the biochemically confirmed S. aureus isolates were molecularly identified using the thermonuclease (nuc) gene. The antibiotic resistance pattern of all the S. aureus isolates was evaluated through the disc diffusion method. Out of 543 milk samples, 310 (57.09%) were positive for SCM. Among the SCM-positive samples, S. aureus was detected in 30.32% (94/310) samples. Out of 94 isolates, 47 (50%) were determined to be multidrug resistant (MDR). Among these MDR isolates, 11 exhibited resistance to Cefoxitin, and hence were classified as methicillin-resistant Staphylococcus aureus (MRSA). The S. aureus isolates showed the highest resistance to Lincomycin (84.04%) followed by Ampicillin (45.74%), while the least resistance was shown to Sulfamethoxazole/Trimethoprim (3.19%) and Gentamycin (6.38%). Polymerase chain reaction (PCR) analysis revealed that 55.31% of the isolates carried blaZ gene, 46.80% carried tetK gene, 17.02% harbored the mecA gene, whereas, aacA-aphD gene was found in 13.82% samples. Our findings revealed a significant level of contamination of milk with S. aureus and half (50%) of the isolates were MDR. The isolated S. aureus harbored various antibiotic resistance genes responsible for the absorbed phenotypic resistance. The alarmingly high prevalence of MDR S. aureus isolates and MRSA strains in these cases possess a serious risk to public health, emphasizes the urgent need to address this issue to protect both human and animal health in Pakistan.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>38753608</pmid><doi>10.1371/journal.pone.0301200</doi><orcidid>https://orcid.org/0000-0002-8192-5800</orcidid><orcidid>https://orcid.org/0000-0001-5729-8037</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2024-05, Vol.19 (5), p.e0301200-e0301200 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_3069287923 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Ampicillin Analysis Animal health Animals Anti-Bacterial Agents - pharmacology Antibiotic resistance Antibiotics Bacteria Bacterial Proteins - genetics Biology and Life Sciences Catalase Catalase test Cattle Cefoxitin Coagulase Coagulase test Cow's milk Dairy farming Dairy farms Development and progression Drug resistance Drug resistance in microorganisms Drug Resistance, Multiple, Bacterial - genetics Female Food contamination Genes Genetic research Gentamicin Immune system Infection Lincomycin Mammary gland Mammary glands Mannitol Mastitis Mastitis, Bovine - epidemiology Mastitis, Bovine - microbiology MecA protein Medicine and Health Sciences Methicillin Microbial Sensitivity Tests Milk Milk - microbiology Milk production Multidrug resistance Nosocomial infections Pakistan - epidemiology Pathogens Plasmids Polymerase chain reaction Proteins Public health Ribonucleic acid RNA Skin Staphylococcal infections Staphylococcal Infections - epidemiology Staphylococcal Infections - microbiology Staphylococcal Infections - veterinary Staphylococcus aureus Staphylococcus aureus - drug effects Staphylococcus aureus - genetics Staphylococcus aureus - isolation & purification Staphylococcus infections Sulfamethoxazole Tetracycline Tetracyclines Thermonuclease Toxins Trimethoprim Virulence Wildlife conservation |
| title | Alarming multidrug resistance in Staphylococcus aureus isolated from raw milk of cows with subclinical mastitis: Antibiotic resistance patterns and occurrence of selected resistance genes |
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