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Previous issue date: 2007-02-05 / The main purpose of this study was to compare genotypically samples of Corynebacterium pseudotuberculosis obtained from the abscesses of sheep and goats diagnosed with caseous lymphadenitis proceeding from the semi-arid region of Pernambuco, Brazil. The RFLP-PCR technique was used to fingerprint the genes rpoB and pld, with the restriction enzymes Hpy-Ch4 and MspI; and PstI and MspI applied, respectively. The banding pattern observed through eletrophoreses was similar in all samples for both genes studied, independently of the restriction enzyme utilized. The in vitro sensitivity profile was also analyzed by testing the samples with 14 different antimicrobials. Initially, the contents of the abscesses were cultivated in blood-agar and the isolated colonies obtained were identified using the API-Coryne biochemical identification kit (Bio-Merieux, France), and selecting Gram-positive and nitrate-negative samples, which were classified as Corynebacterium pseudotuberculosis. The disk diffusion technique was used to perform antibiogramsand the results showed that 96.8% of the samples were sensitive to chloramphenicol and ciprofloxacin; 93.5% were sensitive to norfloxacin and cefazolin; 90.3% to amoxicillin and tetracycline; 87.1% to sulfa-trimethoprim; 83.9% to orbifloxacin; 77.4% to ampicillin, lincomycin and penicillin; 32% to gentamicin; 12.9% to novobiocin and 9.7% to neomycin. It was also noted that 87.1% of the samples were resistant to novobiocin; 80.6% to neomycin; 41.9% to gentamicin; 19.3% to lincomycin; 16.1% to ampicillin, orbifloxacin and penicillin; 12.9% to sulfatrimethoprim; 6.4% to cefazolin, norfloxacin and amoxicillin, and 3.2% to ciprofloxacin, chloramphenicol and tetracycline. For all samples, with the exception of one, was found some degree of multiple resistances with 16% of them being multiresistant to approximately 50% of the antimicrobial drugs tested. Based on those results, it is possible to conclude that there is a phenotypic variation among the samples analyzed, which could be associated to one or more resistance genes. Thegenotypic results did not demonstrate any difference on the band fragmentation pattern among the samples when analyzing the genes pld and rpoB, regardless of the host species origin or geographic area, indicating a homogeneous genotypic profile of the infection in the region of study. / Objetivou-se com este estudo comparar genotipicamente amostras de Corynebacterium pseudotuberculosis recuperadas do conteúdo de abscessos de caprinos e ovinos com linfadenite caseosa procedentes do Sertão de Pernambuco, Brasil. Utilizou-se a técnica de fingerprint RFLP-PCR aplicadas aos genes rpoB e Pld. Os genes foram tratados com Hpy-Ch 4 e Msp I e Pst I e Msp I, respectivamente.Os perfis de restrição observados foram semelhantes entre todas as amostras para os dois genes estudados. Avaliou-se o perfil de sensibilidade in vitro frente a 14 antimicrobianos. Inicialmente o conteúdo dos abscessos foi cultivado em ágar-sangue e as colônias isoladas foram identificadas pelo kit de identificação bioquímico API-Coryne (Bio-Merieux – França), sendo selecionadas as amostras Gram-positivas e nitrato-negativas, classificadas como C. pseudotuberculosis. Para a realização dos antibiogramas, utilizou-se a técnica de difusão em discos. A sensibilidade aos antimicrobianos demonstram que 96,8% das amostras foramsensíveis ao cloranfenicol e ciprofloxacina; 93,5% a norfloxacina e cefazolina; 90,3% a amoxicilina e tetraciclina; 87,1% a sulfa-trimetoprim; 83,9% a orbifloxacina; 77,4% a ampicilina, lincomicina e penicilina; 32% a gentamicina; 12,9% a novobiocina e 9,7% a neomicina. Quanto à resistência, 87,1% das amostras foram resistentes à novobiocina; 80,6% a neomicina; 41,9% a gentamicina; 19,3% a lincomicina; 16,1% a ampicilina, orbifloxacina e penicilina; 12,9% a sulfa-trimetoprim; 6,4% a cefazolina, norfloxacina e amoxicilina e 3,2% a ciprofloxacina, cloranfenicol e tetraciclina. Com exceção de uma amostra todas as demais apresentaram algum grau de resistência múltipla, sendo que 16% delas foram multirresistentes a aproximadamente 50% dos antimicrobianos testados. Concluiu-se que apesar de existir uma variação fenotípica entre as amostras analisadas que pode estar associada a um ou mais genes de resistência, os resultados da genotipagem demonstram não haver qualquer diferença nos padrões de fragmentos de bandas entre as amostras para os genes pld e rpoB, independente da origem da espéciehospedeira ou da área geográfica, indicando um padrão genotípico homogêneo de infecção na região estudada.
| Identifer | oai:union.ndltd.org:IBICT/oai:tede2:tede2/5844 |
| Date | 05 February 2007 |
| Creators | ABREU, Sílvio Romero de Oliveira |
| Contributors | MOTA, Rinaldo Aparecido, SOARES, Cleber Oliveira, LANGONI, Hélio, OLIVEIRA, Andréa Alice da Fonseca, GOMES FILHO, Manoel Adrião, CASTRO, Roberto Soares de |
| Publisher | Universidade Federal Rural de Pernambuco, Programa de Pós-Graduação em Ciência Veterinária, UFRPE, Brasil, Departamento de Medicina Veterinária |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFRPE, instname:Universidade Federal Rural de Pernambuco, instacron:UFRPE |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | -3061482854177903105, 600, 600, 600, -3020210563763616780, 453670264235017319 |
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