Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes

Detalhes bibliográficos
Autor(a) principal: SOARES, Karla Danielle Almeida
Data de Publicação: 2019
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRPE
Texto Completo: http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/8189
Resumo: Dairy production in Brazil stands out as one of the most important activities for Brazilian livestock, being responsible for the source of income of millions of Brazilians. However, this activity is constantly influenced by the occurrence of mastitis, especially in the subclinical form, which occurs in herds. Staphylococcus spp is the main genus of mastitis-causing bacteria in cattle. The aim of this study was to investigate enterotoxigenic Staphylococcus aureus biofilm-forming isolates from bovine mastitis in dairy herds and to profile resistance against disinfectants. Ten farms linked to a federally registered Milk Processing power plant were selected. Physical examination of the mammary gland and milk of the animals was performed and then the California Mastitis Test. Samples were collected from 960 animals for microbiological examination after antisepsis of the roof ostium with 70 ° GL alcohol. In the laboratory, aliquots of 10 μl of milk were seeded on 5% sheep blood agar and then plates incubated at 37 ° C for 48 hours. Subsequently, 213 isolates Staphylococcus spp. classified as strong and moderate biofilm producers were selected for characterization regarding the phenotypic profile and presence of the femA gene for identification of S. aureus. Of these, 152 isolates were confirmed, which were subjected to detection of genes capable of producing biofilms (icaA, icaD and Bap), genes encoding enterotoxins (SeA, SeB, SeC, SeD, SeE), and resistance related genes (Tet (k), Vana). The action of disinfectants (chlorhexidine and lactic acid) used in the milking routine on biofilm in formation and consolidated in 86 of these samples was tested. In 84.16% (134/152) of the samples the BAP gene was found, in 78.29% (119/152) the IcaD gene and in only 1.32% (2/152) the IcaA gene. Only enterotoxins Sec in 17.76% (27/152) of the samples and See in 0.66% (1/152). TetK resistance genes were found in 48.68% (74/152) of the samples. Satisfactory results were observed for the two disinfectants tested on the biofilm in formation, in 94.2% (81/86) of chlorhexidine isolates and 100% of lactic acid isolates. However, for the consolidated biofilm the action percentage was much lower, the lactic acid showed low efficacy over the already formed biofilm, with only 3.5% of reduction rate, while chlorhexidine was able to reduce 43% of the consolidated biofilm. The effect of disinfectants on reducing biofilm adhesion was quite significant. Research into more effective products that act on both free bacterial cells and microorganisms in the form of biofilms is suggested.
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spelling MEDEIROS, Elizabeth Sampaio deNERO, Luís AugustoPINHEIRO JUNIOR, José WiltonROLIM, Maria Betânia de QueirozSILVA, Karla Patrícia Chaves daBARBOSA, Julicelly Gomeshttp://lattes.cnpq.br/5770903127454350SOARES, Karla Danielle Almeida2019-09-10T13:41:40Z2019-07-09SOARES, Karla Danielle Almeida. Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes. 2019. 78 f. Tese (Programa de Pós-Graduação em Biociência Animal) - Universidade Federal Rural de Pernambuco, Recife.http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/8189Dairy production in Brazil stands out as one of the most important activities for Brazilian livestock, being responsible for the source of income of millions of Brazilians. However, this activity is constantly influenced by the occurrence of mastitis, especially in the subclinical form, which occurs in herds. Staphylococcus spp is the main genus of mastitis-causing bacteria in cattle. The aim of this study was to investigate enterotoxigenic Staphylococcus aureus biofilm-forming isolates from bovine mastitis in dairy herds and to profile resistance against disinfectants. Ten farms linked to a federally registered Milk Processing power plant were selected. Physical examination of the mammary gland and milk of the animals was performed and then the California Mastitis Test. Samples were collected from 960 animals for microbiological examination after antisepsis of the roof ostium with 70 ° GL alcohol. In the laboratory, aliquots of 10 μl of milk were seeded on 5% sheep blood agar and then plates incubated at 37 ° C for 48 hours. Subsequently, 213 isolates Staphylococcus spp. classified as strong and moderate biofilm producers were selected for characterization regarding the phenotypic profile and presence of the femA gene for identification of S. aureus. Of these, 152 isolates were confirmed, which were subjected to detection of genes capable of producing biofilms (icaA, icaD and Bap), genes encoding enterotoxins (SeA, SeB, SeC, SeD, SeE), and resistance related genes (Tet (k), Vana). The action of disinfectants (chlorhexidine and lactic acid) used in the milking routine on biofilm in formation and consolidated in 86 of these samples was tested. In 84.16% (134/152) of the samples the BAP gene was found, in 78.29% (119/152) the IcaD gene and in only 1.32% (2/152) the IcaA gene. Only enterotoxins Sec in 17.76% (27/152) of the samples and See in 0.66% (1/152). TetK resistance genes were found in 48.68% (74/152) of the samples. Satisfactory results were observed for the two disinfectants tested on the biofilm in formation, in 94.2% (81/86) of chlorhexidine isolates and 100% of lactic acid isolates. However, for the consolidated biofilm the action percentage was much lower, the lactic acid showed low efficacy over the already formed biofilm, with only 3.5% of reduction rate, while chlorhexidine was able to reduce 43% of the consolidated biofilm. The effect of disinfectants on reducing biofilm adhesion was quite significant. Research into more effective products that act on both free bacterial cells and microorganisms in the form of biofilms is suggested.A produção leiteira no Brasil se destaca por ser uma das atividades mais importantes para pecuária brasileira, sendo responsável pela fonte de renda de milhões de brasileiros. Entretanto, essa atividade é constantemente influenciada pela ocorrência da mastite, principalmente na forma subclínica, que ocorre nos rebanhos. Staphylococcus spp constitui o principal gênero de bactérias causadoras de mastite em bovinos. Objetivou-se com este estudo pesquisar Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e traçar o perfil de resistência frente à desinfetantes. Foram selecionadas 10 fazendas vinculadas à uma Usina de beneficiamento de leite sob registro federal. Foi realizado o exame físico da glândula mamária e do leite dos animais e, em seguida, realizado o California Mastitis Test. Foram coletadas amostras de 960 animais para o exame microbiológico após antissepsia do óstio dos tetos com álcool 70°GL. No laboratório, alíquotas de 10 μL de leite foram semeadas em ágar sangue ovino a 5% e, em seguida, as placas incubadas a 37°C por 48 horas. Posteriormente, 213 isolados Staphylococcus spp. classificados como fortes e moderados produtores de biofilmes foram selecionados para caracterização quanto ao perfil fenotípico e presença do gene femA para identificação de S. aureus. Destes, foram confirmados 152 isolados, que foram submetidos a detecção de genes capazes de produzir biofilmes (icaA, icaD e Bap), genes que codificam as enterotoxinas (SeA, SeB, SeC, SeD, SeE), e genes relacionados a resistência (Tet(k), Vana). Testou-se a ação de desinfetantes (clorexidine e ácido láctico) utilizados na rotina de ordenha sobre o biofilme em formação e consolidado em 86 destas amostras. Em 84,16% (134/152) das amostras foram constatados o gene BAP, em 78,29% (119/152) o gene IcaD e, em apenas 1,32% (2/152) o gene IcaA. Apenas as enterotoxinas Sec em 17,76% (27/152) das amostras e See em 0,66% (1/152). Foram encontrados genes de resistência TetK em 48,68% (74/152) das amostras. Observou-se resultado satisfatório dos dois desinfetantes testados sobre o biofilme em formação, em 94,2% (81/86) dos isolados submetidos ao clorexidine e 100% dos submetidos ao ácido láctico. Entretanto, para o biofilme consolidado o percentual de ação foi bem inferior, o ácido láctico apresentou baixa eficácia sobre o biofilme já formado, com apenas 3,5% da taxa de redução, enquanto a clorexidine conseguiu reduzir 43% do biofilme consolidado. O efeito dos desinfetantes na redução da adesão de biofilmes foi bastante significativo. Sugere-se a investigação de produtos mais eficazes, que atuem tanto nas células bacterianas livres, como nos microrganismos na forma de biofilmes.Submitted by Mario BC (mario@bc.ufrpe.br) on 2019-09-10T13:41:40Z No. of bitstreams: 1 Karla Danielle Almeida Soares.pdf: 621702 bytes, checksum: f0c9b78fb0dce47ff82edcb658f26a3a (MD5)Made available in DSpace on 2019-09-10T13:41:40Z (GMT). No. of bitstreams: 1 Karla Danielle Almeida Soares.pdf: 621702 bytes, checksum: f0c9b78fb0dce47ff82edcb658f26a3a (MD5) Previous issue date: 2019-07-09application/pdfporUniversidade Federal Rural de PernambucoPrograma de Pós-Graduação em Biociência AnimalUFRPEBrasilDepartamento de Morfologia e Fisiologia AnimalStaphylococcus aureusBovino de leiteMastiteDesinfetanteCIENCIAS AGRARIAS::MEDICINA VETERINARIAPesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis-1510757014399315592600600600-8922364187987396204453670264235017319info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFRPEinstname:Universidade Federal Rural de Pernambuco (UFRPE)instacron:UFRPEORIGINALKarla Danielle Almeida Soares.pdfKarla Danielle Almeida Soares.pdfapplication/pdf621702http://www.tede2.ufrpe.br:8080/tede2/bitstream/tede2/8189/2/Karla+Danielle+Almeida+Soares.pdff0c9b78fb0dce47ff82edcb658f26a3aMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82165http://www.tede2.ufrpe.br:8080/tede2/bitstream/tede2/8189/1/license.txtbd3efa91386c1718a7f26a329fdcb468MD51tede2/81892019-09-10 10:41:40.444oai:tede2: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Biblioteca Digital de Teses e Dissertaçõeshttp://www.tede2.ufrpe.br:8080/tede/PUBhttp://www.tede2.ufrpe.br:8080/oai/requestbdtd@ufrpe.br ||bdtd@ufrpe.bropendoar:2024-05-28T12:36:39.096636Biblioteca Digital de Teses e Dissertações da UFRPE - Universidade Federal Rural de Pernambuco (UFRPE)false
dc.title.por.fl_str_mv Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes
title Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes
spellingShingle Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes
SOARES, Karla Danielle Almeida
Staphylococcus aureus
Bovino de leite
Mastite
Desinfetante
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes
title_full Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes
title_fullStr Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes
title_full_unstemmed Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes
title_sort Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes
author SOARES, Karla Danielle Almeida
author_facet SOARES, Karla Danielle Almeida
author_role author
dc.contributor.advisor1.fl_str_mv MEDEIROS, Elizabeth Sampaio de
dc.contributor.advisor-co1.fl_str_mv NERO, Luís Augusto
dc.contributor.referee1.fl_str_mv PINHEIRO JUNIOR, José Wilton
dc.contributor.referee2.fl_str_mv ROLIM, Maria Betânia de Queiroz
dc.contributor.referee3.fl_str_mv SILVA, Karla Patrícia Chaves da
dc.contributor.referee4.fl_str_mv BARBOSA, Julicelly Gomes
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/5770903127454350
dc.contributor.author.fl_str_mv SOARES, Karla Danielle Almeida
contributor_str_mv MEDEIROS, Elizabeth Sampaio de
NERO, Luís Augusto
PINHEIRO JUNIOR, José Wilton
ROLIM, Maria Betânia de Queiroz
SILVA, Karla Patrícia Chaves da
BARBOSA, Julicelly Gomes
dc.subject.por.fl_str_mv Staphylococcus aureus
Bovino de leite
Mastite
Desinfetante
topic Staphylococcus aureus
Bovino de leite
Mastite
Desinfetante
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
dc.subject.cnpq.fl_str_mv CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Dairy production in Brazil stands out as one of the most important activities for Brazilian livestock, being responsible for the source of income of millions of Brazilians. However, this activity is constantly influenced by the occurrence of mastitis, especially in the subclinical form, which occurs in herds. Staphylococcus spp is the main genus of mastitis-causing bacteria in cattle. The aim of this study was to investigate enterotoxigenic Staphylococcus aureus biofilm-forming isolates from bovine mastitis in dairy herds and to profile resistance against disinfectants. Ten farms linked to a federally registered Milk Processing power plant were selected. Physical examination of the mammary gland and milk of the animals was performed and then the California Mastitis Test. Samples were collected from 960 animals for microbiological examination after antisepsis of the roof ostium with 70 ° GL alcohol. In the laboratory, aliquots of 10 μl of milk were seeded on 5% sheep blood agar and then plates incubated at 37 ° C for 48 hours. Subsequently, 213 isolates Staphylococcus spp. classified as strong and moderate biofilm producers were selected for characterization regarding the phenotypic profile and presence of the femA gene for identification of S. aureus. Of these, 152 isolates were confirmed, which were subjected to detection of genes capable of producing biofilms (icaA, icaD and Bap), genes encoding enterotoxins (SeA, SeB, SeC, SeD, SeE), and resistance related genes (Tet (k), Vana). The action of disinfectants (chlorhexidine and lactic acid) used in the milking routine on biofilm in formation and consolidated in 86 of these samples was tested. In 84.16% (134/152) of the samples the BAP gene was found, in 78.29% (119/152) the IcaD gene and in only 1.32% (2/152) the IcaA gene. Only enterotoxins Sec in 17.76% (27/152) of the samples and See in 0.66% (1/152). TetK resistance genes were found in 48.68% (74/152) of the samples. Satisfactory results were observed for the two disinfectants tested on the biofilm in formation, in 94.2% (81/86) of chlorhexidine isolates and 100% of lactic acid isolates. However, for the consolidated biofilm the action percentage was much lower, the lactic acid showed low efficacy over the already formed biofilm, with only 3.5% of reduction rate, while chlorhexidine was able to reduce 43% of the consolidated biofilm. The effect of disinfectants on reducing biofilm adhesion was quite significant. Research into more effective products that act on both free bacterial cells and microorganisms in the form of biofilms is suggested.
publishDate 2019
dc.date.accessioned.fl_str_mv 2019-09-10T13:41:40Z
dc.date.issued.fl_str_mv 2019-07-09
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dc.identifier.citation.fl_str_mv SOARES, Karla Danielle Almeida. Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes. 2019. 78 f. Tese (Programa de Pós-Graduação em Biociência Animal) - Universidade Federal Rural de Pernambuco, Recife.
dc.identifier.uri.fl_str_mv http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/8189
identifier_str_mv SOARES, Karla Danielle Almeida. Pesquisa de Staphylococcus aureus enterotoxigênicos formadores de biofilmes isolados de bovinos com mastite em rebanhos leiteiros e perfil de resistência frente a desinfetantes. 2019. 78 f. Tese (Programa de Pós-Graduação em Biociência Animal) - Universidade Federal Rural de Pernambuco, Recife.
url http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/8189
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dc.publisher.initials.fl_str_mv UFRPE
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dc.publisher.department.fl_str_mv Departamento de Morfologia e Fisiologia Animal
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