PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Costa M.M., Machado S.A., Krewer C.C., Ilha M.R.S., Graça D.L., Guaraldi A.L.M. & Vargas A.C. 2006. Pathogenicity of Rhodococcus equi in mice, isolated from environment, human and horse clinical samples. Pesquisa Veterinária Brasileira 26(3):167-170. Laboratório de Bacteriologia, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Rurais, Universidade Federal de Santa Maria, Avenida Roraima 1000, Santa Maria, RS 97105-900, Brazil. E-mail: agueda@ccr.ufsm.br Rhodococcus equi is a facultative intracellular pathogen associated with bronchopneumonia, mesenteric lymphadenitis and enterocolitis in foals. Although R. equi is likely to be found in every horse-breeding farm, the clinical disease is unrecognized in most of them. Capsule components, equi factor, micolic acid and some products encoded by the large 85-90Kb plasmid were described as virulence factors. However, the pathogenesis of R. equi infections and the sensibility of foals are not completely understood. The aim of this study was evaluate the virulence of R. equi isolated from human, horses and environment for mices. Nine strains carrying the 85-90Kb plasmid isolated from foal clinical specimens, one from immunodeficient human patient and six plasmidless strains (four isolated from feces, one from pasture and one from immunodeficient human patient) were inoculated in cyclophosphamide immunossuppressed mice. The pathological changes and viability of R. equi cells in the liver of mice was verified after the 3rd, 6th an 10th day after inoculation for horse and environmental isolates and for R. equi isolates from human patients on the 1st, 3rd and 6th day. During the necropsy procedures, infiltrate of macrophages and pyogranulomatous lesions were detected after the sixth pos-inoculation day in the liver and spleen. In horse isolates, only plasmid positive strains were virulent, but in human isolates both strains (plasmid positive e plasmid negative) were virulent. Both groups of the immunossupressed mice inoculated with R. equi isolated from environment showed pathological changes. All R. equi strains were unable to kill non imunossuppressed mice.

• Rhodococcus equi mice virulence cyclophosphamide

Abstract in Portuguese:

ABSTRACT.- Costa M.M., Machado S.A., Krewer C.C., Ilha M.R.S., Graça D.L., Guaraldi A.L.M. & Vargas A.C. 2006. Pathogenicity of Rhodococcus equi in mice, isolated from environment, human and horse clinical samples. Pesquisa Veterinária Brasileira 26(3):167-170. Laboratório de Bacteriologia, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Rurais, Universidade Federal de Santa Maria, Avenida Roraima 1000, Santa Maria, RS 97105-900, Brazil. E-mail: agueda@ccr.ufsm.br Rhodococcus equi is a facultative intracellular pathogen associated with bronchopneumonia, mesenteric lymphadenitis and enterocolitis in foals. Although R. equi is likely to be found in every horse-breeding farm, the clinical disease is unrecognized in most of them. Capsule components, equi factor, micolic acid and some products encoded by the large 85-90Kb plasmid were described as virulence factors. However, the pathogenesis of R. equi infections and the sensibility of foals are not completely understood. The aim of this study was evaluate the virulence of R. equi isolated from human, horses and environment for mices. Nine strains carrying the 85-90Kb plasmid isolated from foal clinical specimens, one from immunodeficient human patient and six plasmidless strains (four isolated from feces, one from pasture and one from immunodeficient human patient) were inoculated in cyclophosphamide immunossuppressed mice. The pathological changes and viability of R. equi cells in the liver of mice was verified after the 3rd, 6th an 10th day after inoculation for horse and environmental isolates and for R. equi isolates from human patients on the 1st, 3rd and 6th day. During the necropsy procedures, infiltrate of macrophages and pyogranulomatous lesions were detected after the sixth pos-inoculation day in the liver and spleen. In horse isolates, only plasmid positive strains were virulent, but in human isolates both strains (plasmid positive e plasmid negative) were virulent. Both groups of the immunossupressed mice inoculated with R. equi isolated from environment showed pathological changes. All R. equi strains were unable to kill non imunossuppressed mice.

Abstract in English:

ABSTRACT.- Spilki F.R., Esteves P.A., Franco A.C., Lima M., Holz C.L., Batista H.B.C.R., Driemeier D., Flores E.F., Weiblen R. & Roehe P.M. 2002. [Neurovirulence and neuroinvasiveness of bovine herpesvirus type 1 and 5 in rabbits.] Neurovirulência e neuroinvasividade de Herpesvírus bovinos tipos 1 e 5 em coelhos. Pesquisa Veterinária Brasileira 22(2):58-63. Centro de Pesquisas Veterinárias Desidério Finamor, Estrada do Conde 6000, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@orion.ufrgs.br In order to determine the capacity of bovine herpesvirus type 1 and 5 (BHV-1 and BHV-5) to invade, multiply and spread along the central nervous system (CNS) (neuroinvasiveness), as well as their potential to induce neurological illness (neurovirulence), 30 to 35 days old rabbits were inoculated with the BHV-5 strain EVI 88 / 95 and Los Angeles and Cooper BHV-1 strains, by the intrathecal (IT) and intranasal (IN) routes. The BHV-5 strain induced severe neurological clinical signs in 100% (12/12) of the rabbits inoculated by both routes. Histopathological examination revealed multifocal non-suppurative meningoencephalitis, characterized by multifocal gliosis and perivascular cuffing. Virus was recovered from many parts of the brain. Both BHV-1 strains, when inoculated via 1T route, were not neurovirulent. The strain Los Angeles, after IN inoculation, induced signs of severe respiratory disease (7/7), as well as signs of neurological impairment, indistinguishable from those induced by BHV-5, in 57% (4/7) of the infected rabbits. However, the rabbits with nervous signs revealed at histopathology vasculitis and thrombosis in lungs and brain, the latter with foci of neuronal necrosis, but no lesions indicative of encephalitis, suggesting that neural damage was probably consequent to tissue anoxia. The BHV-1 strain Cooper, after IN inoculation, induced only mild signs of respiratory disease. These findings indicate that the BHV-5 strain was both neuroinvasive and neurovirulent, since it was capable of invading, spreading and multiplying in the rabbits brains by both routes of inoculation, yet causing neurological disease, apparently consequent to vírus induced neural damage. The BHV-1 Los Angeles strain was not neuroinvasive, whereas its neurovirulence was probably consequent to tissue anoxia, which histologically seemed not to be related to direct viral pathogenic effect. The BHV-1 strain Cooper was neither neurovirulent nor neuroinvasive for rabbits. It is possible that these observations bear relationship with the frequent association of BHV-5 with encephalitis in cattle, as opposed to BHV-1 encephalitis, which is a rare event in nature.

• Infectious bovine rhinotracheitis IBR BHV-1 bovine encephalitis herpesvirus BHV-5 Rinotraqueíte infecciosa bovina herpesvírus da encefalite bovina.

Abstract in Portuguese:

RESUMO.- Spilki F.R., Esteves P.A., Franco A.C., Lima M., Holz C.L., Batista H.B.C.R., Driemeier D., Flores E.F., Weiblen R. & Roehe P.M. 2002. [Neurovirulence and neuroinvasiveness of bovine herpesvirus type 1 and 5 in rabbits.] Neurovirulência e neuroinvasividade de Herpesvírus bovinos tipos 1 e 5 em coelhos. Pesquisa Veterinária Brasileira 22(2):58-63. Centro de Pesquisas Veterinárias Desidério Finamor, Estrada do Conde 6000, Cx. Postal 47, Eldorado do Sul, RS 92990-000, Brazil. E-mail: proehe@orion.ufrgs.br Com o objetivo de avaliar a capacidade dos herpesvírus bovinos tipos 1 e 5 (BHV-1 e BHV-5) de invadir e replicar no sistema nervoso central (SNC) (neuroinvasividade), bem como sua capacidade de induzir doença neurológica (neurovirulência), coelhos com 30 a 35 dias de idade foram inoculados com uma amostra do Herpesvírus da Encefalite Bovina (BHV-5; amostra EVI 88/95) ou com amostras de BHV-1 (Los Angeles ou Cooper), pelas vias intratecal (IT) e intranasal (IN). A inoculação da amostra de BHV-5, tanto pela via 1T como IN, induziu sinais clínicos neurológicos em 100% (12/12) dos coelhos inoculados. Os exames histopatológicos revelaram um quadro de meningoencefalite não-purulenta multifocal, caracterizada por gliose multifocal e infiltrados perivasculares. O vírus foi isolado de várias áreas do SNC desses animais. As amostras de BHV-1, quando inoculadas pela via IT, não foram neurovirulentas. A amostra Los Angeles de BHV-1, quando administrada pela via IN, induziu sinais respiratórios severos, além de sinais neurológicos em 57% (4/7) dos animais inoculados. Entretanto, o exame histopatológico destes quatro animais revelou vasculite e trombose no pulmão e cérebro, este último apresentando focos de necrose neuronal, porém sem lesões indicativas de encefalite. Isso sugere que os sinais neurológicos foram, provavelmente, consequentes a prejuízos no fluxo sangüíneo encefálico, e não a danos neuronais provocados pela inoculação desse vírus. A amostra Cooper de BHV-1, quando inoculada pela via IN, induziu apenas sinais leves de infecção respiratória. Estes resultados indicam que apenas a amostra de BHV-5 foi capaz de invadir e replicar no encéfalo dos coelhos quando inoculada tanto por via IN como IT, apresentando neuroinvasividade e neurovirulência. É possível que estas observações tenham relação com o fato de amostras de BHV-5 freqüentemente causarem encefalites, em contraposição a infecções pelo BHV-1, onde encefalites são raramente observadas.

Abstract in English:

Virulence factors were determined in 43 cultures of Yersinia enterocolitica serotype 0:3, biotype 4, phagetype VIII, isolated from apparently healthy pigs. Twenty one isolates were from tonsils, twelve from feces, six from colon contents and four from tongues. The in vitro tests consisted of examining autoagglutination at 36ºC, and absorption of Congo Red. The virulence assays of Y. enterocolitica were conducted by in vivo tests, testing the invasiveness into guinea-pig eyes and detection of heat stable enterotoxin in suckling mice. The results revealed a higher positivity percentage of the autoagglutination test (76.7%), followed by absorption of Congo Red (53.5%), invasiveness into guinea-pig eyes (20.9%) and detection of heat-stable enterotoxin (7.0%). A higher correlation was found in vitro test as compareci to in vivo tests, which characterized 53.5% Y. enterocolitica isolates tested as virulent and which allowed the conclusion that the simultaneous utilization of autoagglutination and absorption of Congo Red tests may be relevant for the identification of pathogenic strains.

• Yersinia enterocolitica pathogenicity swine

Abstract in Portuguese:

Foram pesquisados os fatores de virulência em 43 estirpes de Yersinia enterocolitica sorotipo 0:3, biotipo 4, lisotipo VIII isoladas de suínos sadios, sendo 21 amostras provenientes de amigdala, 12 de fezes, seis de conteúdo de cólon e quatro de língua. A avaliação da virulência foi verificada pela invasibilidade em olho de cobaia, detecção de enterotoxina termoestável em camundongo lactente, auto aglutinação a 36 ºC e absorção do vermelho Congo. Os resultados revelaram um maior percentual de positividade no teste de auto-aglutinação (76.7%), seguido da absorção do vermelho Congo (53,9%), invasibilidade em olho de cobaia (20,9%) e detecção de enterotoxina termoestável (7,0%). Evidenciou-se maior correlação entre os testes realizados "in vitro" quando comparados às provas "in vivo", caracterizando 53,5% das amostra testadas, como virulentas. A utilização simultânea dos testes de auto-aglutinação a 36°C e absorção do vermelho Congo na identificação de cepas patogênicas parecem ser importantes.

• Yersinia enterocolitica patogenicidade suíno

Abstract in English:

Experimental neonatal colibacillosis, in newbom piglets was attempted using 4 groups of enterotoxigenic Escherichia coli (ETEC) strains, as follows: 1) Two strains from serogroup 0149:K91, both producing thermolabile enterotoxin (LT) and K88 colonization factor; 2) Two strains from serogroup 0101:K30, producing thermostable enterotoxin (STa) and K99 colonization factor; 3) One strain from serogroup 0157:K?, producing thermostable enterotoxin of the STb type and K88 antigen, and 4) One strain from serogroup 08:K?, producing STa enterotoxin anda new colonization factor, named F42. All fourteen piglets inoculated orally with these strains of ETEC developed clinical disease and died up to 42 hours after inoculation, being possible to visualize, by indirect fluorescent antibody technique, in all of them, that colonization of small intestine by the inoculated ETEC had occurred. The production of STa "in vivo", into the gut, by strains from group 2 and 4 was an important factor to prove that experimental colibacillosis did occur. In fact, coprocultures either from the diarrheic stools or from the gut contents revealed a high rate of LT+-K88+ and STa+ -K99+ colonies recovery. Though some quantitative differences among the examined materiais have been observed, the recovery of STa + -F42 + colonies was lower than in the former groups of ETEC strains. However clinical symptoms, production of STa "in vivo" and colonization of the gut of inoculated piglets proved that F42 antigen is undoubtedly a new colonization factor among ETEC involved in porcine colibacillosis.

• Porcine colibacillosis experimental reproduction virulence factors LT STa K88 K99 and F42

Abstract in Portuguese:

Foi tentada a reprodução experimental da colibacilose suína neonatal, em leitões recém-nascidos, usando-se para tal 4 grupos de amostras de Escherichia coli enterotoxigênicas (ETEC), a saber: 1) Duas amostras do sorogrupo 0149:K81, produtoras da enterotoxina termolábil (L T) e do fator de colonização K88; 2) Duas amostras do sorogrupo 0101:K30, produtoras da enterotoxina termoestável (STa) e do fator de colonização K99; 3) Uma amostra do sorogrupo 0157:K?, produtora da enterotoxina termoestável do tipo STb e do fator de colonização K88, e 4) Uma amostra do sorogrupo 08:K?, produtora da enterotoxina termoestável (STa) e de um novo fator de colonização, denominado F42. Todos os 14 leitões inoculados por via oral com estas amostras de ETEC desenvolveram doença clínica com morte até 42 horas após a inoculação, tendo sido possível detectar em todos eles a colonização do intestino delgado pelas amostras de ETEC inoculadas, através da técnica de imunofluorescência indireta. A produção de STa "in vivo", por amostras dos grupos 2 e 4 foi um fator importante na comprovação de que a reprodução experimental da doença por estas amostras realmente ocorreu. De fato, a coprocultura, quer das fezes diarréicas, quer do conteúdo intestinal dos animais, revelou um alto índice de recuperação de colonias LT+ -K88+ e STa+ -K99+. Embora tenham ocorrido entre os diversos materiais examinados algumas diferenças quantitativas, a recuperação de colônias STa=+ -F42+ foi menor do que nos casos anteriores, porém os achados referentes a doença clínica, produção de STa "in vivo" e colonização do intestino delgado dos leitões inoculados, comprovaram que o antígeno F42 é, sem dúvida, um novo fator de colonização em amostras de ETEC envolvidas na colibacilose suína.

• Colibacilose suína reprodução experimental fatores de virulência LT STa K88 K99 e F42

Abstract in English:

Four hundred and seventy-seven cultures of Escherichia coli isolated in the region of Concórdia, Santa Catarina, Brazil, were examined for the presence of ''virulence factors", such as: thermolabile (LT) enterotoxin; thermostable (STa and STb) enterotoxins and the colonization factors K88, K99 and 987P. Paired samples of sera from some sick piglets· were also collected for the search of anti-LT antibodies. Eighty-six (18,02%) E. coli cultures produced ·STa enterotoxin whereas LT enterotoxin was detected in 8 (1,67%) only. Among 391 STa- cultures 49 (12,53%) were STb+. Among 28 K88+ cultures, 1 produced LT; 9 STa and the remaining were non-enterotoxigenic. None of the L T+ cultures codified for K99 antigen, which, on the other hand, was found in 5 STa+ and in 17 non-entérotoxigenic cultures. 987P antigen was found in 3 non-enterotoxigenic cultures. Four out of 10 samples of the examined paired sera showed serological conversion in relation to titres found in ·the passive immunehemolysis (PIH) test. The results of serogrouping revealed that all LT+ cultures belonged to serogroup 01.49. Among 86 STa+ cultures, 1 was grouped as serogroup 09, 2 as 010, 4 as 035; 2 as 064; 3 as 0108; 2 as 0138; 1 as 0149 and 1 as 0157. Seventy STa+ cultures were not classified.· Among 49 STb+ cultures 1 belonged to serogroup 09; 4 to 010; 1 to 035; 1 to 0139; 2 to 0149; 1 to 0157 and 39 cultures could not be serogrouped. These results suggest that in porcine colibacillosis of the region of Concórdia, SC, Brasil the thermostable enterotoxins (STa and STb) may play an important role. It could be possible that among these cultures other colonization factors, different from K88, K99 and 987P may occur. Also, most cultures examined could not be grouped among the serogroups generally accepted as enteropathogenic for swine.

• Virulence factors porcine colibacillosis Concórdia Brazil

Abstract in Portuguese:

Quatrocentos e setenta e sete amostras de Escherichia coli, isoladas na região de Concórdia, SC, Brazil, foram examinadas quanto a presença de fatores de virulência a saber: enterotoxína termolábil (LT), enterotoxinas termoestáveis (STa e STb) e os fatores de colonização K88, K99 e 987P. Amostras pareadas de soro de alguns leitões doentes foram também coletadas para a pesquisa de anticorpos anti-LT. Oitenta e seis (18,02%) amostras de E. coli produziram a enterotoxina STa enquanto a enterotoxina LT-foi detectada em apenas 8 (1.67%). Entre as-381 amostras STa-, 49 (12.53%) foram STb+. Entre 28 amostras K88+, uma produziu LT, 9 STa e as restantes eram não enterotoxigenicas. Nenhuma das amostras ·LT+ codificou para o antígeno K99, porém, este foi encontrado em 5 amostras STa+ e em 17 não entérotoxigênicas. Quatro entre os 10 soros examinados apresentaram conversão sorológica nos níveis de anticorpos anti-LT quando os soros foram examinados pelo teste da imunohemólise passiva (PIH). Os resultados dos exames sorológicos revelaram que todas as amostras LT+ pertenciam ao sorogrupo 0149. Entre 86 amostras STa+, 1 foi classificada como pertencendo ao sorogrupo 09, 2 ao 010; 4 ao 035; 2 ao 064; 3 ao 0108; 2 ao 0138; 1 ao 0149; 1 ao 0157 e 70 amostras não foram Classificadas. Entre as 49 amostras STb+ 1 pertencia ao sorogrupo 09; 4 ao 010; 1 ao 035; 1 ao 0139; 2 ao 0149; 1 ao 0157 e 39 não foram classificadas. Estes resultados sugerem que na colibacilose suína da região· de Concórdia as enterotoxinas termoestáveis (STa e STb) possam desempenhar um importante papel, existindo a possibilidade de que nelas possam ocorrer outros fatores de colonização que não K88, K99 e 987P. As amostras estudadas também, em sua maioria, não se enquadraram entre· aqueles sorogrupos geralmente aceitos como enteropatogênicos para suínos.

• Fatores de virulência colibacilose suína Concórdia Brazil