Comparison phenotypic and genotypic identification of Staphylococcus species isolated from bovine mastitis
In addition to Staphylococcus aureus nowadays other coagulase-positive staphylococci (CoPS) and coagulase-negative staphylococci (CoNS), earlier considered of minor importance, are now accepted as relevant pathogens for humans and animals. The involvement of these microorganisms in bovine mastitis e...
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description | In addition to Staphylococcus aureus nowadays other coagulase-positive staphylococci (CoPS) and coagulase-negative staphylococci (CoNS), earlier considered of minor importance, are now accepted as relevant pathogens for humans and animals. The involvement of these microorganisms in bovine mastitis etiology and the possibility their transmission through milk to humans justify the requirement of developing reliable methods for identification of the most frequent species among them. The purpose of this study was to compare the phenotypic techniques with the genotypic method carried out by sequencing of the rpoB gene in identification of several species of the genus Staphylococcus isolated from bovine mastitis. A total of 300 staphylococci isolates of bovine mastitis cases from several Brazilian dairy herds were studied by phenotypic and genotypic techniques, respectively: 150 CoPS and 150 CoNS strains. A total of 18 CoNS different species and 4 CoPS species were identified. Among the CoNS the following species were recognized: 48 (32%) Staphylococcus warneri, 22(15%) S. epidermidis, 20(13%) S. hyicus, 10(7%) S. xylosus, 7(5%) S. haemolyticus, 6(4%) S. simulans, 6(4%) S. schleiferi subsp schleiferi, 6(4%) S. hominis, 5(3%) S. pasteuri, 4(2.7%) S. cohnii, 3(2%) S. saprophyticus subsp. saprophyticus 3(2%) S. chromogenes 3(2%) S. sciuri, 2(1%) S. saccharolyticus, 2(1%) S. lugdunensi, 1(0,7%) S. auricularis, 1(70%) S. saprophyticus subsp. bovis, 1(0.7%) S. capitis. And among the 150 CoPS were identified respectively: 105 (70%) S. aureus, 21(14%), S. hyicus, 19(13%) S. intermedius e 5(3%) S. schleiferi subsp coagulans. Considering the 150 CoNS isolates, the identifications performed by phenotypic and genotypic tests presented 96.7% of concordance, kappa coefficient of agreement = 0.933, SE (standard error) of kappa=0.021 (95% confidence interval: 0.893 to 0.974), Pearson’s correlation coefficient (r) = 0.9977, (confidence interval 95%: 0.9938 a 0.9992) and in relation to 150 CPS isolates it was detected an agreement of 98.7%, kappa = 0.960, SE of kappa = 0.016, (95% confidence interval: 0.929 to 0.992) Pearson’s correlation coefficient (r) = 0.9994 (95% confidence interval: 0.9681 to 1.0000). The verified agreement strength between the identification methods can be considered as excellent. These results assure that according to laboratory resources any of them will be suitable to perform the staphylococci identification.
RESUMO: Além de Staphylococcus aureus atualm |
doi_str_mv | 10.1590/s0100-736x2016001200003 |
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RESUMO: Além de Staphylococcus aureus atualmente outros estafilococos coagulase positiva (SCP) e estafilococos coagulase-negativos (SCN), anteriormente considerados de menor relevância, são reconhecidos como importantes patógenos para humanos e animais. O envolvimento desses micro-organismos na etiologia da mastite bovina e a possibilidade da sua transmissão através do leite aos humanos justifica a utilização de métodos confiáveis para a identificação das espécies mais frequentes. O objetivo deste estudo foi comparar as técnicas fenotípicas com o método genotípico realizada por sequenciamento do gene rpoB na identificação de espécies do gênero Staphylococcus spp. isolados de mastite bovina. Um total de 300 estafilococos isolados de casos de mastite bovina em diferentes rebanhos leiteiros brasileiros foram estudados por técnicas fenotípicas e genotípicas, respectivamente: 150 linhagens de SCP e 150 linhagens de SCN. Foram identificados um total de 18 espécies de SCN e 4 espécies SCP. Entre os SCN as seguintes espécies identificadas: 48 (32%) Staphylococcus warneri, 22 (15%) S. epidermidis, 20 (13%) S. hyicus, 10 (7%) S. xylosus, 7 (5%) S. haemolyticus, 6 (4%) S. simulans, 6 (4%) S. schleiferi subsp schleiferi, 6 (4%) S. hominis, 5 (3%) S. pasteuri, 4 (2,7%) S. cohnii, 3 (2%) S. saprophyticus subsp. saprophyticus, 3 (2%) S. chromogenes, 3 (2%) S. sciuri, 2 (1%) S. saccharolyticus, 2 (1%) S. lugdunensi, 1 (0,7%) S. auricularis, 1 (70 %) S. saprophyticus subsp. bovis, 1 (0,7%) S. capitis. E entre as 150 SCP foram identificados, 105 (70%) S. aureus, 21 (14%), S. hyicus, 19 (13%) S. intermedius e 5 (3%) S. schleiferi subsp coagulans. Considerando-se os 150 SCN isolados, as identificações realizadas por testes fenotípicos e genotípicos apresentaram 96,7% de concordância, coeficiente de concordância kappa = 0,933, SE (erro padrão) de kappa = 0,021 (95% intervalo de confiança: 0,893-0,974), coeficiente de correlação de Pearson (r) = 0,9977, (intervalo de confiança de 95%: 0,9938 a 0,9992) e em relação a 150 SCP isolados foi observado uma concordância de 98,7%, kappa = 0,960, sE de kappa = 0,016, (95% de intervalo de confiança: 0,929 a 0,992) coeficiente de correlação de Pearson (r) = 0,9994 (95% intervalo de confiança: 0,9681-1,0000). A correlação entre os métodos de identificação pode ser considerada como excelente. Esses resultados demonstraram que de acordo com os recursos disponíveis no laboratório, poderia ser utilizada qualquer uma das metodologias.</description><identifier>ISSN: 0100-736X</identifier><identifier>ISSN: 1678-5150</identifier><identifier>EISSN: 0100-736X</identifier><identifier>DOI: 10.1590/s0100-736x2016001200003</identifier><language>eng</language><publisher>Colégio Brasileiro de Patologia Animal - CBPA</publisher><subject>biochemical tests ; coagulase negative Staphylococcus ; coagulase positive Staphylococcus ; genotype ; mastitis ; Phenotype ; rpoB gene ; sequencing ; Staphylococcus spp ; VETERINARY SCIENCES</subject><ispartof>Pesquisa Veterinária Brasileira, 2016-12, Vol.36 (12), p.1160-1164</ispartof><rights>This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c411t-d32fa589bdfb15fb6b722670d5dfacea883a03f6d4963c01ea693e06c2ce9bb83</citedby><cites>FETCH-LOGICAL-c411t-d32fa589bdfb15fb6b722670d5dfacea883a03f6d4963c01ea693e06c2ce9bb83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,864,885,27924,27925</link.rule.ids></links><search><creatorcontrib>Guimarães, Felipe Freitas</creatorcontrib><creatorcontrib>Joaquim, Samea Fernandes</creatorcontrib><creatorcontrib>Manzi, Marcela Pinho</creatorcontrib><creatorcontrib>Silva, Rodrigo Costa da</creatorcontrib><creatorcontrib>Bruder-Nascimento, Ariane Cristina Mendes de Oliveira</creatorcontrib><creatorcontrib>Costa, Elizabeth Oliveira</creatorcontrib><creatorcontrib>Langoni, Hélio</creatorcontrib><title>Comparison phenotypic and genotypic identification of Staphylococcus species isolated from bovine mastitis</title><title>Pesquisa Veterinária Brasileira</title><addtitle>Pesq. Vet. Bras</addtitle><description>In addition to Staphylococcus aureus nowadays other coagulase-positive staphylococci (CoPS) and coagulase-negative staphylococci (CoNS), earlier considered of minor importance, are now accepted as relevant pathogens for humans and animals. The involvement of these microorganisms in bovine mastitis etiology and the possibility their transmission through milk to humans justify the requirement of developing reliable methods for identification of the most frequent species among them. The purpose of this study was to compare the phenotypic techniques with the genotypic method carried out by sequencing of the rpoB gene in identification of several species of the genus Staphylococcus isolated from bovine mastitis. A total of 300 staphylococci isolates of bovine mastitis cases from several Brazilian dairy herds were studied by phenotypic and genotypic techniques, respectively: 150 CoPS and 150 CoNS strains. A total of 18 CoNS different species and 4 CoPS species were identified. Among the CoNS the following species were recognized: 48 (32%) Staphylococcus warneri, 22(15%) S. epidermidis, 20(13%) S. hyicus, 10(7%) S. xylosus, 7(5%) S. haemolyticus, 6(4%) S. simulans, 6(4%) S. schleiferi subsp schleiferi, 6(4%) S. hominis, 5(3%) S. pasteuri, 4(2.7%) S. cohnii, 3(2%) S. saprophyticus subsp. saprophyticus 3(2%) S. chromogenes 3(2%) S. sciuri, 2(1%) S. saccharolyticus, 2(1%) S. lugdunensi, 1(0,7%) S. auricularis, 1(70%) S. saprophyticus subsp. bovis, 1(0.7%) S. capitis. And among the 150 CoPS were identified respectively: 105 (70%) S. aureus, 21(14%), S. hyicus, 19(13%) S. intermedius e 5(3%) S. schleiferi subsp coagulans. Considering the 150 CoNS isolates, the identifications performed by phenotypic and genotypic tests presented 96.7% of concordance, kappa coefficient of agreement = 0.933, SE (standard error) of kappa=0.021 (95% confidence interval: 0.893 to 0.974), Pearson’s correlation coefficient (r) = 0.9977, (confidence interval 95%: 0.9938 a 0.9992) and in relation to 150 CPS isolates it was detected an agreement of 98.7%, kappa = 0.960, SE of kappa = 0.016, (95% confidence interval: 0.929 to 0.992) Pearson’s correlation coefficient (r) = 0.9994 (95% confidence interval: 0.9681 to 1.0000). The verified agreement strength between the identification methods can be considered as excellent. These results assure that according to laboratory resources any of them will be suitable to perform the staphylococci identification.
RESUMO: Além de Staphylococcus aureus atualmente outros estafilococos coagulase positiva (SCP) e estafilococos coagulase-negativos (SCN), anteriormente considerados de menor relevância, são reconhecidos como importantes patógenos para humanos e animais. O envolvimento desses micro-organismos na etiologia da mastite bovina e a possibilidade da sua transmissão através do leite aos humanos justifica a utilização de métodos confiáveis para a identificação das espécies mais frequentes. O objetivo deste estudo foi comparar as técnicas fenotípicas com o método genotípico realizada por sequenciamento do gene rpoB na identificação de espécies do gênero Staphylococcus spp. isolados de mastite bovina. Um total de 300 estafilococos isolados de casos de mastite bovina em diferentes rebanhos leiteiros brasileiros foram estudados por técnicas fenotípicas e genotípicas, respectivamente: 150 linhagens de SCP e 150 linhagens de SCN. Foram identificados um total de 18 espécies de SCN e 4 espécies SCP. Entre os SCN as seguintes espécies identificadas: 48 (32%) Staphylococcus warneri, 22 (15%) S. epidermidis, 20 (13%) S. hyicus, 10 (7%) S. xylosus, 7 (5%) S. haemolyticus, 6 (4%) S. simulans, 6 (4%) S. schleiferi subsp schleiferi, 6 (4%) S. hominis, 5 (3%) S. pasteuri, 4 (2,7%) S. cohnii, 3 (2%) S. saprophyticus subsp. saprophyticus, 3 (2%) S. chromogenes, 3 (2%) S. sciuri, 2 (1%) S. saccharolyticus, 2 (1%) S. lugdunensi, 1 (0,7%) S. auricularis, 1 (70 %) S. saprophyticus subsp. bovis, 1 (0,7%) S. capitis. E entre as 150 SCP foram identificados, 105 (70%) S. aureus, 21 (14%), S. hyicus, 19 (13%) S. intermedius e 5 (3%) S. schleiferi subsp coagulans. Considerando-se os 150 SCN isolados, as identificações realizadas por testes fenotípicos e genotípicos apresentaram 96,7% de concordância, coeficiente de concordância kappa = 0,933, SE (erro padrão) de kappa = 0,021 (95% intervalo de confiança: 0,893-0,974), coeficiente de correlação de Pearson (r) = 0,9977, (intervalo de confiança de 95%: 0,9938 a 0,9992) e em relação a 150 SCP isolados foi observado uma concordância de 98,7%, kappa = 0,960, sE de kappa = 0,016, (95% de intervalo de confiança: 0,929 a 0,992) coeficiente de correlação de Pearson (r) = 0,9994 (95% intervalo de confiança: 0,9681-1,0000). A correlação entre os métodos de identificação pode ser considerada como excelente. Esses resultados demonstraram que de acordo com os recursos disponíveis no laboratório, poderia ser utilizada qualquer uma das metodologias.</description><subject>biochemical tests</subject><subject>coagulase negative Staphylococcus</subject><subject>coagulase positive Staphylococcus</subject><subject>genotype</subject><subject>mastitis</subject><subject>Phenotype</subject><subject>rpoB gene</subject><subject>sequencing</subject><subject>Staphylococcus spp</subject><subject>VETERINARY SCIENCES</subject><issn>0100-736X</issn><issn>1678-5150</issn><issn>0100-736X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>DOA</sourceid><recordid>eNpVkd1uGyEQhVdVIuWneYbwAnYG8LK7l5XVNpYi9SKJ1Ds0C0PMar2sgFT12xfHtdVyMzBwPmbOVNU9hyWvO3hIwAEWjVS_BXAFwAWUJT9V16eLnxf_7K-qm5QGANHIhl9XwzrsZow-hYnNW5pC3s_eMJwsezufvKUpe-cNZl_eBceeM87b_RhMMOY9sTST8ZRYwYyYyTIXw4714ZefiO0wZZ99-lxdOhwT3f2Nt9Xrt68v68fF04_vm_WXp4VZcZ4XVgqHddv11vW8dr3qGyFUA7a2Dg1h20oE6ZRddUoa4ISqkwTKCENd37fyttocuTbgoOfodxj3OqDXH4kQ3zTG7M1IurO2JklCNAW3sqItfzoEVQtCV3dUWMsjK5X-xqCH8B6nUrx-PhiqD4aeXeclFkFzFJgYUorkzgVw0Id56XRS_j8v-Qf_y4q8</recordid><startdate>20161201</startdate><enddate>20161201</enddate><creator>Guimarães, Felipe Freitas</creator><creator>Joaquim, Samea Fernandes</creator><creator>Manzi, Marcela Pinho</creator><creator>Silva, Rodrigo Costa da</creator><creator>Bruder-Nascimento, Ariane Cristina Mendes de Oliveira</creator><creator>Costa, Elizabeth Oliveira</creator><creator>Langoni, Hélio</creator><general>Colégio Brasileiro de Patologia Animal - CBPA</general><general>Colégio Brasileiro de Patologia Animal (CBPA)</general><scope>AAYXX</scope><scope>CITATION</scope><scope>GPN</scope><scope>DOA</scope></search><sort><creationdate>20161201</creationdate><title>Comparison phenotypic and genotypic identification of Staphylococcus species isolated from bovine mastitis</title><author>Guimarães, Felipe Freitas ; Joaquim, Samea Fernandes ; Manzi, Marcela Pinho ; Silva, Rodrigo Costa da ; Bruder-Nascimento, Ariane Cristina Mendes de Oliveira ; Costa, Elizabeth Oliveira ; Langoni, Hélio</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c411t-d32fa589bdfb15fb6b722670d5dfacea883a03f6d4963c01ea693e06c2ce9bb83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>biochemical tests</topic><topic>coagulase negative Staphylococcus</topic><topic>coagulase positive Staphylococcus</topic><topic>genotype</topic><topic>mastitis</topic><topic>Phenotype</topic><topic>rpoB gene</topic><topic>sequencing</topic><topic>Staphylococcus spp</topic><topic>VETERINARY SCIENCES</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Guimarães, Felipe Freitas</creatorcontrib><creatorcontrib>Joaquim, Samea Fernandes</creatorcontrib><creatorcontrib>Manzi, Marcela Pinho</creatorcontrib><creatorcontrib>Silva, Rodrigo Costa da</creatorcontrib><creatorcontrib>Bruder-Nascimento, Ariane Cristina Mendes de Oliveira</creatorcontrib><creatorcontrib>Costa, Elizabeth Oliveira</creatorcontrib><creatorcontrib>Langoni, Hélio</creatorcontrib><collection>CrossRef</collection><collection>SciELO</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>Pesquisa Veterinária Brasileira</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Guimarães, Felipe Freitas</au><au>Joaquim, Samea Fernandes</au><au>Manzi, Marcela Pinho</au><au>Silva, Rodrigo Costa da</au><au>Bruder-Nascimento, Ariane Cristina Mendes de Oliveira</au><au>Costa, Elizabeth Oliveira</au><au>Langoni, Hélio</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparison phenotypic and genotypic identification of Staphylococcus species isolated from bovine mastitis</atitle><jtitle>Pesquisa Veterinária Brasileira</jtitle><addtitle>Pesq. Vet. Bras</addtitle><date>2016-12-01</date><risdate>2016</risdate><volume>36</volume><issue>12</issue><spage>1160</spage><epage>1164</epage><pages>1160-1164</pages><issn>0100-736X</issn><issn>1678-5150</issn><eissn>0100-736X</eissn><abstract>In addition to Staphylococcus aureus nowadays other coagulase-positive staphylococci (CoPS) and coagulase-negative staphylococci (CoNS), earlier considered of minor importance, are now accepted as relevant pathogens for humans and animals. The involvement of these microorganisms in bovine mastitis etiology and the possibility their transmission through milk to humans justify the requirement of developing reliable methods for identification of the most frequent species among them. The purpose of this study was to compare the phenotypic techniques with the genotypic method carried out by sequencing of the rpoB gene in identification of several species of the genus Staphylococcus isolated from bovine mastitis. A total of 300 staphylococci isolates of bovine mastitis cases from several Brazilian dairy herds were studied by phenotypic and genotypic techniques, respectively: 150 CoPS and 150 CoNS strains. A total of 18 CoNS different species and 4 CoPS species were identified. Among the CoNS the following species were recognized: 48 (32%) Staphylococcus warneri, 22(15%) S. epidermidis, 20(13%) S. hyicus, 10(7%) S. xylosus, 7(5%) S. haemolyticus, 6(4%) S. simulans, 6(4%) S. schleiferi subsp schleiferi, 6(4%) S. hominis, 5(3%) S. pasteuri, 4(2.7%) S. cohnii, 3(2%) S. saprophyticus subsp. saprophyticus 3(2%) S. chromogenes 3(2%) S. sciuri, 2(1%) S. saccharolyticus, 2(1%) S. lugdunensi, 1(0,7%) S. auricularis, 1(70%) S. saprophyticus subsp. bovis, 1(0.7%) S. capitis. And among the 150 CoPS were identified respectively: 105 (70%) S. aureus, 21(14%), S. hyicus, 19(13%) S. intermedius e 5(3%) S. schleiferi subsp coagulans. Considering the 150 CoNS isolates, the identifications performed by phenotypic and genotypic tests presented 96.7% of concordance, kappa coefficient of agreement = 0.933, SE (standard error) of kappa=0.021 (95% confidence interval: 0.893 to 0.974), Pearson’s correlation coefficient (r) = 0.9977, (confidence interval 95%: 0.9938 a 0.9992) and in relation to 150 CPS isolates it was detected an agreement of 98.7%, kappa = 0.960, SE of kappa = 0.016, (95% confidence interval: 0.929 to 0.992) Pearson’s correlation coefficient (r) = 0.9994 (95% confidence interval: 0.9681 to 1.0000). The verified agreement strength between the identification methods can be considered as excellent. These results assure that according to laboratory resources any of them will be suitable to perform the staphylococci identification.
RESUMO: Além de Staphylococcus aureus atualmente outros estafilococos coagulase positiva (SCP) e estafilococos coagulase-negativos (SCN), anteriormente considerados de menor relevância, são reconhecidos como importantes patógenos para humanos e animais. O envolvimento desses micro-organismos na etiologia da mastite bovina e a possibilidade da sua transmissão através do leite aos humanos justifica a utilização de métodos confiáveis para a identificação das espécies mais frequentes. O objetivo deste estudo foi comparar as técnicas fenotípicas com o método genotípico realizada por sequenciamento do gene rpoB na identificação de espécies do gênero Staphylococcus spp. isolados de mastite bovina. Um total de 300 estafilococos isolados de casos de mastite bovina em diferentes rebanhos leiteiros brasileiros foram estudados por técnicas fenotípicas e genotípicas, respectivamente: 150 linhagens de SCP e 150 linhagens de SCN. Foram identificados um total de 18 espécies de SCN e 4 espécies SCP. Entre os SCN as seguintes espécies identificadas: 48 (32%) Staphylococcus warneri, 22 (15%) S. epidermidis, 20 (13%) S. hyicus, 10 (7%) S. xylosus, 7 (5%) S. haemolyticus, 6 (4%) S. simulans, 6 (4%) S. schleiferi subsp schleiferi, 6 (4%) S. hominis, 5 (3%) S. pasteuri, 4 (2,7%) S. cohnii, 3 (2%) S. saprophyticus subsp. saprophyticus, 3 (2%) S. chromogenes, 3 (2%) S. sciuri, 2 (1%) S. saccharolyticus, 2 (1%) S. lugdunensi, 1 (0,7%) S. auricularis, 1 (70 %) S. saprophyticus subsp. bovis, 1 (0,7%) S. capitis. E entre as 150 SCP foram identificados, 105 (70%) S. aureus, 21 (14%), S. hyicus, 19 (13%) S. intermedius e 5 (3%) S. schleiferi subsp coagulans. Considerando-se os 150 SCN isolados, as identificações realizadas por testes fenotípicos e genotípicos apresentaram 96,7% de concordância, coeficiente de concordância kappa = 0,933, SE (erro padrão) de kappa = 0,021 (95% intervalo de confiança: 0,893-0,974), coeficiente de correlação de Pearson (r) = 0,9977, (intervalo de confiança de 95%: 0,9938 a 0,9992) e em relação a 150 SCP isolados foi observado uma concordância de 98,7%, kappa = 0,960, sE de kappa = 0,016, (95% de intervalo de confiança: 0,929 a 0,992) coeficiente de correlação de Pearson (r) = 0,9994 (95% intervalo de confiança: 0,9681-1,0000). A correlação entre os métodos de identificação pode ser considerada como excelente. Esses resultados demonstraram que de acordo com os recursos disponíveis no laboratório, poderia ser utilizada qualquer uma das metodologias.</abstract><pub>Colégio Brasileiro de Patologia Animal - CBPA</pub><doi>10.1590/s0100-736x2016001200003</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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identifier | ISSN: 0100-736X |
ispartof | Pesquisa Veterinária Brasileira, 2016-12, Vol.36 (12), p.1160-1164 |
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language | eng |
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source | DOAJ Directory of Open Access Journals; EZB-FREE-00999 freely available EZB journals |
subjects | biochemical tests coagulase negative Staphylococcus coagulase positive Staphylococcus genotype mastitis Phenotype rpoB gene sequencing Staphylococcus spp VETERINARY SCIENCES |
title | Comparison phenotypic and genotypic identification of Staphylococcus species isolated from bovine mastitis |
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