Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial

Detalhes bibliográficos
Autor(a) principal: Moraes, Dunya Mara Cardoso
Data de Publicação: 2014
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFG
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/6460
Resumo: The purpose of this research was to investigate presence of Salmonella sp. by conventional bacterial identification and real-time PCR on eggshell, albumen, and yolk of washed and unwashed commercial white and brown eggs; in samples of flooring material from transport crates (meconium); raising environment (swab of cages and drinking fountains); cloacal swab; food and insects from growing, rearing and production phases in a commercial group of laying hens. Also, was examinated the susceptibility profile of Salmonella isolates to 14 antimicrobials: sulfamethoxazole (25μg), trimethoprim-sulfamethoxazole (25μg), enrofloxacin (5μg), tetracyclin (30μg) sulphonamides (300μg), ciprofloxacin (10μg), amoxicillin and clavulanic acid (3μg), ampicillin (20μg), ceftiofur (30μg), gentamicin (10μg), oxytetracycline (30μg), neomycin (30μg), doxycycline (30μg) and apramycin (15μg); and evaluated the presence of virulence gene spvC and resistence genes intl1, sul1 and blaTEM. A total of 68 dozens of white eggs and 61 dozens of brown eggs were randomly collected in poultry houses and 30 dozens and 34 dozens of white and brown eggs, respectively, were collected in egg-storage room (on the farm). Each collected dozen corresponded to three sample units: eggshell, albumen and yolk, totaling 387 samples. Salmonella spp. was detected by conventional bacteriology in 5.4% of analyzed samples and in 16% by qPCR. Salmonella Agona represented 18.2% of identified serovars, Salmonella enterica subs. enterica O:4.5 18.2%, Salmonella Schwarzengrund 18.2%, Salmonella Cerro 13.6%, Salmonella Anatum 13.6%, Salmonella Enteritidis 9.1%, Salmonella Johannesburg 4.5%, Salmonella Corvallis 4.5%, Salmonella Livingstone and Salmonella Senftenberg. From 864 samples collected (248 samples originated from growing, 392 from rearing and 224 from production phase), 2,8% where positives in bacteriologic technique and 15,3% in qPCR. Contamination was higher in growing and rearing phases and declined in production phase. Twenty four isolations of Salmonella where typified as Salmonella Agona (41,7%), Salmonella Livingstone (33,3%), Salmonella Cerro (16,7%), Salmonella Senftenberg (4,2%) and Salmonella Schwarzengrund (4,2%). The highest resistance percentage observed were to sulfamethoxazole (91,0%), sulphonamides (51%) and ceftiofur (28,9%) and 0% to ciprofloxacin. Only Salmonella Johannesburg and Salmonella Corvallis showed resistance to only one antibiotic and others serovars were resistant to at least two antimicrobials, noting that Salmonella Schwarzengrund presented resistance to 13 of them. The gene spvC was dectected only in serovar Enteritidis, while intl1 was present in six of ten analyzed serovars and the gene sul1 in three of them, always in association with intl1. The gene blaTEM was not identified. In conclusion, this research points numerous serovars circulating in commercial egg structures and phenotypic antimicrobials resistance investigation, along with antimicrobial resistance genes in isolates of Salmonella sp. are important investigation tools to determinate genetic profile of these bacteria.
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spelling Andrade, Maria Auxiliadorahttp://lattes.cnpq.br/9441751521255467Jayme, Valéria de Sáhttp://lattes.cnpq.br/0603234425928309Andrade, Maria AuxiliadoraStringhini, Maria LuizaDuarte, Sabrina CastilhoLinhares, Guido Fontgalland CoelhoMesquita, Albenones José dehttp://lattes.cnpq.br/9632713129112672Moraes, Dunya Mara Cardoso2016-11-08T17:37:11Z2014-04-11MORAES, D. M. C. Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial. 2014. 95 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás,Goiânia, 2014.http://repositorio.bc.ufg.br/tede/handle/tede/6460The purpose of this research was to investigate presence of Salmonella sp. by conventional bacterial identification and real-time PCR on eggshell, albumen, and yolk of washed and unwashed commercial white and brown eggs; in samples of flooring material from transport crates (meconium); raising environment (swab of cages and drinking fountains); cloacal swab; food and insects from growing, rearing and production phases in a commercial group of laying hens. Also, was examinated the susceptibility profile of Salmonella isolates to 14 antimicrobials: sulfamethoxazole (25μg), trimethoprim-sulfamethoxazole (25μg), enrofloxacin (5μg), tetracyclin (30μg) sulphonamides (300μg), ciprofloxacin (10μg), amoxicillin and clavulanic acid (3μg), ampicillin (20μg), ceftiofur (30μg), gentamicin (10μg), oxytetracycline (30μg), neomycin (30μg), doxycycline (30μg) and apramycin (15μg); and evaluated the presence of virulence gene spvC and resistence genes intl1, sul1 and blaTEM. A total of 68 dozens of white eggs and 61 dozens of brown eggs were randomly collected in poultry houses and 30 dozens and 34 dozens of white and brown eggs, respectively, were collected in egg-storage room (on the farm). Each collected dozen corresponded to three sample units: eggshell, albumen and yolk, totaling 387 samples. Salmonella spp. was detected by conventional bacteriology in 5.4% of analyzed samples and in 16% by qPCR. Salmonella Agona represented 18.2% of identified serovars, Salmonella enterica subs. enterica O:4.5 18.2%, Salmonella Schwarzengrund 18.2%, Salmonella Cerro 13.6%, Salmonella Anatum 13.6%, Salmonella Enteritidis 9.1%, Salmonella Johannesburg 4.5%, Salmonella Corvallis 4.5%, Salmonella Livingstone and Salmonella Senftenberg. From 864 samples collected (248 samples originated from growing, 392 from rearing and 224 from production phase), 2,8% where positives in bacteriologic technique and 15,3% in qPCR. Contamination was higher in growing and rearing phases and declined in production phase. Twenty four isolations of Salmonella where typified as Salmonella Agona (41,7%), Salmonella Livingstone (33,3%), Salmonella Cerro (16,7%), Salmonella Senftenberg (4,2%) and Salmonella Schwarzengrund (4,2%). The highest resistance percentage observed were to sulfamethoxazole (91,0%), sulphonamides (51%) and ceftiofur (28,9%) and 0% to ciprofloxacin. Only Salmonella Johannesburg and Salmonella Corvallis showed resistance to only one antibiotic and others serovars were resistant to at least two antimicrobials, noting that Salmonella Schwarzengrund presented resistance to 13 of them. The gene spvC was dectected only in serovar Enteritidis, while intl1 was present in six of ten analyzed serovars and the gene sul1 in three of them, always in association with intl1. The gene blaTEM was not identified. In conclusion, this research points numerous serovars circulating in commercial egg structures and phenotypic antimicrobials resistance investigation, along with antimicrobial resistance genes in isolates of Salmonella sp. are important investigation tools to determinate genetic profile of these bacteria.Objetivou-se com esta pesquisa investigar a presença de Salmonella sp por bacteriologia por bacteriologia por bacteriologia por bacteriologia por bacteriologia por bacteriologia por bacteriologia por bacteriologia por bacteriologia por bacteriologia por bacteriologia por bacteriologia convencional convencionalconvencionalconvencionalconvencionalconvencional convencionalconvencional e PCR em tempo real em casca, albúmen e gema de ovos comerciais brancos e vermelhos, lavados e não lavados; em amostras de forros de caixa de transporte, amostras de ambientes de criação (suabes de gaiola e bebedouro), suabes de cloaca, ração e insetos oriundos das fases de cria, recria e produção de um lote de poedeiras comerciais e investigar o perfil de suscetibilidade de isolados de Salmonella frente 14 antimicrobianos sulfametoxazol (25μg), sulfametoxazol-trimetoprim (25μg), enrofloxacina (5μg), tetraciclina (30μg), sulfonamidas (300μg), ciprofloxacina (10μg), amoxilina ácido clavulânico (3μg), ampicilina (20μg), ceftiofur (30μg), gentamicina (10μg), oxitetraciclina (30μg), neomicina (30μg), doxiciclina (30μg) e apramicina (15μg) e avaliar a presença do gene de virulência spvC e dos genes de resistência intl1, sul1 e blaTEM. Foram colhidas, nos galpões, 68 dúzias de ovos brancos e 27 dúzias de vermelhos e nos entrepostos, após classificação e sanitização, 30 e 34 de dúzias de ovos brancos e vermelhos, respectivamente. Cada dúzia de ovos correspondeu a três amostras: uma amostra de pool casca, uma de pool de albúmen e uma de pool gema, totalizando 387 amostras. Obteve-se pela bacteriologia convencional 5,4% (21/387) e PCR em tempo real 16,0% (62/387) de amostras positivas para Salmonella. Os sorovares identificados foram Salmonella Agona, Salmonella enterica subs. enterica O:4,5, Salmonella Schwarzengrund, Salmonella Cerro, Salmonella Anatum, Salmonella Enteritidis, Salmonella Johannesburg e Salmonella Corvallis. Foram coletadas também 864 amostras no fluxo de produção de ovos e Salmonella sp. foi isolada pela bacteriologia convencional em 2,8% das amostras e pelo PCR em tempo real em 15,3%, sendo que 248 amostras foram originadas da cria, 392 na recria e 224 na produção. De acordo com a analise estatística a infecção evoluiu da fase de cria para a recria e ao chegar a fase de produção o nível de infecção declinou. Os 24 isolados de Salmonella foram tipificados e os sorovares identificados foram Salmonella Agona, Salmonella Livingstone, Salmonella Cerro, Salmonella Senftenberg e Salmonella Schwarzengrund. Foram analisadas também 45 isolados de Salmonella pertencentes aos sorovares Agona, Livingstone, Cerro, Schwarzengrund, Salmonella enterica subs. enterica O:4,5, Anatum, Enteritidis, Johannesburg, Corvallis e Senftenberg. Os maiores percentuais de resistência foram para sulfametoxazol (91,0%), sulfonamidas (51,0%) e ceftiofur (28,9%). O gene spvC foi detectado somente no sorovar Enteritidis, enquanto o gene intl1 esteve presente em seis dos dez sorovares analisados, o gene sul1 em três deles sempre em associação com intl1 e o gene blaTEM não foi identificado. Conclui-se com essa pesquisa que existe uma variedade de sorovares circulantes em granjas de postura e em ovos comerciais; a investigação fenotípica de resistência aos antimicrobianos aliada a pesquisa dos genes de virulência e de resistência aos antibióticos em isolados de Salmonella sp. circulantes em granjas de postura comercial são importante ferramenta de investigação para determinação do perfil genético dessas bactérias. 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dc.title.por.fl_str_mv Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial
dc.title.alternative.eng.fl_str_mv Bacteriological and molecular salmonella sp. investigation at commercial laying houses
title Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial
spellingShingle Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial
Moraes, Dunya Mara Cardoso
Antimicrobianos
genes de virulência
ovos
PCR em tempo real
Antimicrobials
Eggs
PCR
Virulence gene
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial
title_full Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial
title_fullStr Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial
title_full_unstemmed Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial
title_sort Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial
author Moraes, Dunya Mara Cardoso
author_facet Moraes, Dunya Mara Cardoso
author_role author
dc.contributor.advisor1.fl_str_mv Andrade, Maria Auxiliadora
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/9441751521255467
dc.contributor.advisor-co1.fl_str_mv Jayme, Valéria de Sá
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/0603234425928309
dc.contributor.referee1.fl_str_mv Andrade, Maria Auxiliadora
dc.contributor.referee2.fl_str_mv Stringhini, Maria Luiza
dc.contributor.referee3.fl_str_mv Duarte, Sabrina Castilho
dc.contributor.referee4.fl_str_mv Linhares, Guido Fontgalland Coelho
dc.contributor.referee5.fl_str_mv Mesquita, Albenones José de
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/9632713129112672
dc.contributor.author.fl_str_mv Moraes, Dunya Mara Cardoso
contributor_str_mv Andrade, Maria Auxiliadora
Jayme, Valéria de Sá
Andrade, Maria Auxiliadora
Stringhini, Maria Luiza
Duarte, Sabrina Castilho
Linhares, Guido Fontgalland Coelho
Mesquita, Albenones José de
dc.subject.por.fl_str_mv Antimicrobianos
genes de virulência
ovos
PCR em tempo real
topic Antimicrobianos
genes de virulência
ovos
PCR em tempo real
Antimicrobials
Eggs
PCR
Virulence gene
CIENCIAS AGRARIAS::MEDICINA VETERINARIA
dc.subject.eng.fl_str_mv Antimicrobials
Eggs
PCR
Virulence gene
dc.subject.cnpq.fl_str_mv CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description The purpose of this research was to investigate presence of Salmonella sp. by conventional bacterial identification and real-time PCR on eggshell, albumen, and yolk of washed and unwashed commercial white and brown eggs; in samples of flooring material from transport crates (meconium); raising environment (swab of cages and drinking fountains); cloacal swab; food and insects from growing, rearing and production phases in a commercial group of laying hens. Also, was examinated the susceptibility profile of Salmonella isolates to 14 antimicrobials: sulfamethoxazole (25μg), trimethoprim-sulfamethoxazole (25μg), enrofloxacin (5μg), tetracyclin (30μg) sulphonamides (300μg), ciprofloxacin (10μg), amoxicillin and clavulanic acid (3μg), ampicillin (20μg), ceftiofur (30μg), gentamicin (10μg), oxytetracycline (30μg), neomycin (30μg), doxycycline (30μg) and apramycin (15μg); and evaluated the presence of virulence gene spvC and resistence genes intl1, sul1 and blaTEM. A total of 68 dozens of white eggs and 61 dozens of brown eggs were randomly collected in poultry houses and 30 dozens and 34 dozens of white and brown eggs, respectively, were collected in egg-storage room (on the farm). Each collected dozen corresponded to three sample units: eggshell, albumen and yolk, totaling 387 samples. Salmonella spp. was detected by conventional bacteriology in 5.4% of analyzed samples and in 16% by qPCR. Salmonella Agona represented 18.2% of identified serovars, Salmonella enterica subs. enterica O:4.5 18.2%, Salmonella Schwarzengrund 18.2%, Salmonella Cerro 13.6%, Salmonella Anatum 13.6%, Salmonella Enteritidis 9.1%, Salmonella Johannesburg 4.5%, Salmonella Corvallis 4.5%, Salmonella Livingstone and Salmonella Senftenberg. From 864 samples collected (248 samples originated from growing, 392 from rearing and 224 from production phase), 2,8% where positives in bacteriologic technique and 15,3% in qPCR. Contamination was higher in growing and rearing phases and declined in production phase. Twenty four isolations of Salmonella where typified as Salmonella Agona (41,7%), Salmonella Livingstone (33,3%), Salmonella Cerro (16,7%), Salmonella Senftenberg (4,2%) and Salmonella Schwarzengrund (4,2%). The highest resistance percentage observed were to sulfamethoxazole (91,0%), sulphonamides (51%) and ceftiofur (28,9%) and 0% to ciprofloxacin. Only Salmonella Johannesburg and Salmonella Corvallis showed resistance to only one antibiotic and others serovars were resistant to at least two antimicrobials, noting that Salmonella Schwarzengrund presented resistance to 13 of them. The gene spvC was dectected only in serovar Enteritidis, while intl1 was present in six of ten analyzed serovars and the gene sul1 in three of them, always in association with intl1. The gene blaTEM was not identified. In conclusion, this research points numerous serovars circulating in commercial egg structures and phenotypic antimicrobials resistance investigation, along with antimicrobial resistance genes in isolates of Salmonella sp. are important investigation tools to determinate genetic profile of these bacteria.
publishDate 2014
dc.date.issued.fl_str_mv 2014-04-11
dc.date.accessioned.fl_str_mv 2016-11-08T17:37:11Z
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dc.identifier.citation.fl_str_mv MORAES, D. M. C. Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial. 2014. 95 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás,Goiânia, 2014.
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identifier_str_mv MORAES, D. M. C. Investigação bacteriológica e molecular de salmonella sp. em granjas de postura comercial. 2014. 95 f. Tese (Doutorado em Ciência Animal) - Universidade Federal de Goiás,Goiânia, 2014.
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