PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Abreu S.R.O., Mota R.A., Rosinha G.M.S., Forner O., Pinheiro Júnior J.W., Pereira R.R.B., Castro R.S., Elisei C., Soares C.S., Araújo F.R. & Madureira R.C. 2008. [Genotypic comparison between Corynebacterium pseudotuberculosis samples obtained from sheep and goats with caseous lymphadenitis, raised in the semi-arid region of Pernambuco.] Comparação genotípica de isolados de Corynebacterium pseudotuberculosis de caprinos e ovinos do sertão de Pernambuco. Pesquisa Veterinária Brasileira 28(10):481-487. Clínica Escola de Medicina Veterinária, Faculdade de Ciências Biológicas e da Saúde, Centro de Ensino Superior de Maceió, Rodovia Divaldo Suruagy s/n, Quadra 4, Lote 4, Praia do Francês, Marechal Deodoro, AL 57160-000, Brazil. E-mail: silviobiotec@yahoo.com.br The objective was to genotypically compare 35 samples of Corynebacterium pseudotuberculosis obtained from abscesses of sheep and goats diagnosed with caseous lymphadenitis originated from 5 different municipalities in the semi-arid region of Pernambuco, Brazil. The RFLP-PCR technique with Hpy-Ch4 and Msp I and Pst I Msp I restriction enzimes was used to fingerprint the genes rpoB and pld, respectively. The results demonstrate that there was no difference on the fragments banding pattern among samples, independently of the host species or geographic area studied, defining a homogeneous profile of C. pseudotuberculosis responsible for superficial abscesses for the region.

• Genotyping RFLP-PCR Corynebacterium pseudotuberculosis goats sheep

Abstract in Portuguese:

ABSTRACT.- Abreu S.R.O., Mota R.A., Rosinha G.M.S., Forner O., Pinheiro Júnior J.W., Pereira R.R.B., Castro R.S., Elisei C., Soares C.S., Araújo F.R. & Madureira R.C. 2008. [Genotypic comparison between Corynebacterium pseudotuberculosis samples obtained from sheep and goats with caseous lymphadenitis, raised in the semi-arid region of Pernambuco.] Comparação genotípica de isolados de Corynebacterium pseudotuberculosis de caprinos e ovinos do sertão de Pernambuco. Pesquisa Veterinária Brasileira 28(10):481-487. Clínica Escola de Medicina Veterinária, Faculdade de Ciências Biológicas e da Saúde, Centro de Ensino Superior de Maceió, Rodovia Divaldo Suruagy s/n, Quadra 4, Lote 4, Praia do Francês, Marechal Deodoro, AL 57160-000, Brazil. E-mail: silviobiotec@yahoo.com.br The objective was to genotypically compare 35 samples of Corynebacterium pseudotuberculosis obtained from abscesses of sheep and goats diagnosed with caseous lymphadenitis originated from 5 different municipalities in the semi-arid region of Pernambuco, Brazil. The RFLP-PCR technique with Hpy-Ch4 and Msp I and Pst I Msp I restriction enzimes was used to fingerprint the genes rpoB and pld, respectively. The results demonstrate that there was no difference on the fragments banding pattern among samples, independently of the host species or geographic area studied, defining a homogeneous profile of C. pseudotuberculosis responsible for superficial abscesses for the region.

Abstract in English:

ABSTRACT.- Silva M.S., Brum M.C.S., Weiblen R. & Flores E.F. 2007. [Identification and differentiation of herpesvirus types 1 and 5 isolated from clinical samples in central-southern Brazil, Argentina and Uruguay (1987-2006).] Identificação e diferenciação de herpesvírus bovino tipos 1 e 5 isolados de amostras clínicas no Centro-Sul do Brasil, Argentina e Uruguai (1987-2006). Pesquisa Veterinária Brasileira 27(10):403-408. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: flores@ccr.ufsm.br Bovine herpesviruses types 1 and 5 (BoHV-1; BoHV-5) are genetically and antigenically closely related such they can not be distinguished by routine diagnostic tests. As BoHV-1 has been historically associated with respiratory and genital disease, herpesviruses isolated from these clinical syndromes have been tentatively – and sometimes definitively - diagnosed as BoHV-1. Likewise, cases of herpetic neurological infection in cattle have been generally attributed to BoHV-5. This study reports the identification of 40 herpesvirus isolates from different clinical specimens and syndromes in central-southern Brazil, Argentina and Uruguay (1987-2006) by the use of a PCR able to differentiate between BoHV-1 and BoHV-5. BoHV-1 isolates (n=16) were identified in cases of respiratory disease (n=3), vulvovaginitis and/or balanoposthitis (n=3), in semen of healthy bulls (n=5) and in cases of neurological disease (n=5). Viruses identified as BoHV-5 (n=24) were isolated predominantly from cases of neurological disease (n=21), but also from semen of healthy bulls (n=2) and from a spleen of a calf with systemic disease (n=1). These results show that both BoHV-1 and BoHV-5 are not strictly associated with their respective diseases; yet are frequently involved in clinical conditions otherwise attributed to the other virus. These findings also reinforce the need of correctly identifying the herpesvirus isolates as to better understand their pathogenesis and epidemiology.

• Bovine herpesvirus BoHV-1 BoHV-5 differential diagnosis epidemiology

Abstract in Portuguese:

ABSTRACT.- Silva M.S., Brum M.C.S., Weiblen R. & Flores E.F. 2007. [Identification and differentiation of herpesvirus types 1 and 5 isolated from clinical samples in central-southern Brazil, Argentina and Uruguay (1987-2006).] Identificação e diferenciação de herpesvírus bovino tipos 1 e 5 isolados de amostras clínicas no Centro-Sul do Brasil, Argentina e Uruguai (1987-2006). Pesquisa Veterinária Brasileira 27(10):403-408. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. E-mail: flores@ccr.ufsm.br Bovine herpesviruses types 1 and 5 (BoHV-1; BoHV-5) are genetically and antigenically closely related such they can not be distinguished by routine diagnostic tests. As BoHV-1 has been historically associated with respiratory and genital disease, herpesviruses isolated from these clinical syndromes have been tentatively – and sometimes definitively - diagnosed as BoHV-1. Likewise, cases of herpetic neurological infection in cattle have been generally attributed to BoHV-5. This study reports the identification of 40 herpesvirus isolates from different clinical specimens and syndromes in central-southern Brazil, Argentina and Uruguay (1987-2006) by the use of a PCR able to differentiate between BoHV-1 and BoHV-5. BoHV-1 isolates (n=16) were identified in cases of respiratory disease (n=3), vulvovaginitis and/or balanoposthitis (n=3), in semen of healthy bulls (n=5) and in cases of neurological disease (n=5). Viruses identified as BoHV-5 (n=24) were isolated predominantly from cases of neurological disease (n=21), but also from semen of healthy bulls (n=2) and from a spleen of a calf with systemic disease (n=1). These results show that both BoHV-1 and BoHV-5 are not strictly associated with their respective diseases; yet are frequently involved in clinical conditions otherwise attributed to the other virus. These findings also reinforce the need of correctly identifying the herpesvirus isolates as to better understand their pathogenesis and epidemiology.

Abstract in English:

ABSTRACT.- Pereira C.S., Amorim S.D., Santos A.F.M., Siciliano S., Moreno I.M.B., Ott P.H. & Rodrigues D.P. 2007. [Vibrio spp. isolated from marine mammals captured in coastal regions from southwestern to southern Brazil.] Vibrio spp. isolados de mamíferos marinhos capturados na região litorânea do sudeste ao sul do Brasil. Pesquisa Veterinária Brasileira 27(2):81-83. Laboratório de Enterobactérias, Departamento de Bacteriologia, Pavilhão Rocha Lima, Instituto Oswaldo Cruz, FIOCruz, Av. Brasil 4365, Rio de Janeiro, RJ 21045-900, Brazil. E-mail: csoarespereira@hotmail.com In the present investigation was evaluated the incidence of Vibrio spp. from superficial lesions at marine mammals beached or captured by fishing net in the southwestern (RJ) and southern (RS) coastal regions of Brazil. One hundred and ninety eight swabs were collected by DEENSP, GEMARS and Ceclimar institutes and sent to Labent/IOC/FIOCruz where the samples were submitted to enrichment in Alkaline Peptone Water (APW) added with 1% and 3% of sodium chloride (NaCl) incubated at 37oC for 18-24 hours. After the samples were streaked onto Thiossulfate Citrate Bile Sucrose Agar (TCBS), the suspected colonies were submitted to biochemical characterization. The results showed 108 strains, and Vibrio alginolyticus, V. parahaemolyticus, V. vulnificus and V. fluvialis were the main pathogens isolated. These results appoint the importance of surveillance and microbiological monitoring accomplishment and reinforcement of environmental protective programs applied to marine mammals endangered with extinction.

• Vibrionaceae aquatic ecosystem public health

Abstract in Portuguese:

ABSTRACT.- Pereira C.S., Amorim S.D., Santos A.F.M., Siciliano S., Moreno I.M.B., Ott P.H. & Rodrigues D.P. 2007. [Vibrio spp. isolated from marine mammals captured in coastal regions from southwestern to southern Brazil.] Vibrio spp. isolados de mamíferos marinhos capturados na região litorânea do sudeste ao sul do Brasil. Pesquisa Veterinária Brasileira 27(2):81-83. Laboratório de Enterobactérias, Departamento de Bacteriologia, Pavilhão Rocha Lima, Instituto Oswaldo Cruz, FIOCruz, Av. Brasil 4365, Rio de Janeiro, RJ 21045-900, Brazil. E-mail: csoarespereira@hotmail.com In the present investigation was evaluated the incidence of Vibrio spp. from superficial lesions at marine mammals beached or captured by fishing net in the southwestern (RJ) and southern (RS) coastal regions of Brazil. One hundred and ninety eight swabs were collected by DEENSP, GEMARS and Ceclimar institutes and sent to Labent/IOC/FIOCruz where the samples were submitted to enrichment in Alkaline Peptone Water (APW) added with 1% and 3% of sodium chloride (NaCl) incubated at 37oC for 18-24 hours. After the samples were streaked onto Thiossulfate Citrate Bile Sucrose Agar (TCBS), the suspected colonies were submitted to biochemical characterization. The results showed 108 strains, and Vibrio alginolyticus, V. parahaemolyticus, V. vulnificus and V. fluvialis were the main pathogens isolated. These results appoint the importance of surveillance and microbiological monitoring accomplishment and reinforcement of environmental protective programs applied to marine mammals endangered with extinction.

Abstract in English:

Hofer E. & Reis C.M.F. 2005. [Species and serovars of Listeria isolated from sick and clinically healthy animals in Brazil.] Espécies e sorovares de Listeria isolados de animais doentes e portadores no Brasil. Pesquisa Veterinária Brasileira 25(2):79-83. Laboratório de Zoonoses Bacterianas, Depto Bacteriologia, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ 21045-900, Brazil. E-mail: ehofer@ioc.fiocruz.br Two hundred fourty-six strains of the genus Listeria were isolated from sick and clinically healthy animals, collected in three different regions of Brazil during 1971-2000. About 88.2% (217 cultures) yielded Listeria species from faecal specimens of healthy cattle and 29 strains (11.7%) were isolated from sick animals: 15 (6.0%) from central nervous system (CNS) and 14(5.6%) were from otherwise sterile sites. Phenotyping techniques were used to characterize the Listeria isolates. The commonest were L. innocua 6a and non-typable (140/56.9%), L. monocytogenes 4a (37/15.0%) and 4b (22/8.9%), originated mainly from stools of healthy cattle. From sick animals the predominant species and serovars were L. monocytogenes 4b (14/5.6%), and the higher incidence was observed in ruminants (12/4.8%) and 8/3.2% of the serovar 1a from other animal species (rodents and canines) mainly isolated from CNS samples.

• Listeria species serovars sick and healthy animals.

Abstract in Portuguese:

Hofer E. & Reis C.M.F. 2005. [Species and serovars of Listeria isolated from sick and clinically healthy animals in Brazil.] Espécies e sorovares de Listeria isolados de animais doentes e portadores no Brasil. Pesquisa Veterinária Brasileira 25(2):79-83. Laboratório de Zoonoses Bacterianas, Depto Bacteriologia, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ 21045-900, Brazil. E-mail: ehofer@ioc.fiocruz.br Two hundred fourty-six strains of the genus Listeria were isolated from sick and clinically healthy animals, collected in three different regions of Brazil during 1971-2000. About 88.2% (217 cultures) yielded Listeria species from faecal specimens of healthy cattle and 29 strains (11.7%) were isolated from sick animals: 15 (6.0%) from central nervous system (CNS) and 14(5.6%) were from otherwise sterile sites. Phenotyping techniques were used to characterize the Listeria isolates. The commonest were L. innocua 6a and non-typable (140/56.9%), L. monocytogenes 4a (37/15.0%) and 4b (22/8.9%), originated mainly from stools of healthy cattle. From sick animals the predominant species and serovars were L. monocytogenes 4b (14/5.6%), and the higher incidence was observed in ruminants (12/4.8%) and 8/3.2% of the serovar 1a from other animal species (rodents and canines) mainly isolated from CNS samples.

Abstract in English:

ABSTRACT: Guimarães A.M., Lima J.D. & Ribeiro M.EB. 2003. Ultrastructure of Babesia equitrophozoites isolated in Minas Gerais, Brazil. Pesquisa Veterinária Brasileira 23(3):101-104. [Ultra-estrutura de trofozoítos de Babesia equi isolados em Minas Gerais, Brasil.] Departamento de Medicina Veterinária, Universidade Federal de Lavras, Cx. Postal 37, Lavras, MG 37200-000, Brazil. E-mail: amg@ufla.br A transmission electron microscope study was carried out on Babesia equi obtained from a splenectomized horse, from the municipality of Santa Luzia, Minas Gerais, Brazil. The isolate was inoculated into two splenectomized foals (1.05 x 1010 parasitized erythrocytes by B. equi). Trophozoites have a single membrane in direct contact with the cytoplasm of the red blood cells, a prominent nucleus, well-developed rough and smooth endoplasmic reticulum, numerous free ribosomes and small food vacuóles. B. equi trophozoites have a cytostome and a long tubular feeding structure in direct contact with the blood plasmá.

• Babesia equi ultra-estrutura trofozoítos mecanismos de alimentação ultrastructure trophozoites feeding mechanism.

Abstract in Portuguese:

RESUMO: Guimarães A.M., Lima J.D. & Ribeiro M.EB. 2003. Ultrastructure of Babesia equitrophozoites isolated in Minas Gerais, Brazil. Pesquisa Veterinária Brasileira 23(3):101-104. [Ultra-estrutura de trofozoítos de Babesia equi isolados em Minas Gerais, Brasil.] Departamento de Medicina Veterinária, Universidade Federal de Lavras, Cx. Postal 37, Lavras, MG 37200-000, Brazil. E-mail: amg@ufla.br Neste estudo de microsco-pia eletrônica de transmissão utilizou-se um isolado de Babesia equi obtido a partir de um eqüino esplenectomizado, oriundo do município de Santa Luzia, Minas Gerais, Brasil. O isolado foi inoculado em dois potros esplenectomizados (1,05 x 1010 eritrócitos parasitados com B. equi). Os trofozoítos apresentaram uma membrana simples em· contato direto com o citoplasma das· hemácias, núcleo proeminente, retículo endoplasmático liso e rugoso bem desenvolvidos, numerosos ribosomos livres e pequenos vacúolos alimentares. Em trofozoítos de B. equi foi observado citostoma e uma longa estrutura tubular de alimentação em contato direto com o plasma sangüíneo.

Abstract in English:

ABSTRACT.- Mendonça C.L., Vieira D., Kohayagawa A., Schenk M.A.M., Madruga C.R. & Afonso J.A.B. 2003. [Clinical and hematological evaluation of Nelore calves experimentally infected with isolates of Babesia bigemina from the Southeastern, Northeastern and Northern regions of Brazil.] Avaliação clínica e hematológica em bezerros Nelore infectados experimentalmente com isolados de Babesia bigemina das regiões Sudeste, Nordeste e Norte do Brasil. Pesquisa Veterinária Brasileira 23(2):52-60. Clínica de Bovinos, Campus Garanhuns, Univ. Fed. Rural de Pernambuco, Cx. Postal 152, Garanhuns, PE 55292-901, Brazil. A comparative study was made regarding the clinical and hematological alterations caused by isolates of Babesia bigemina from southeastern, northeastern and northern Brazil in experimentally infected Nelore calves. Eighteen calves between 7 and 9 months of age, without antibodies against Babesia sp and raised free from ticks, were used. Three animais were previously inoculated with 2.0x109 parasitic erythrocytes (PE) for each stabilate. The other 15 calves were subdivided into three groups, with five animais each, that were subinoculated with 1.0x1010 PE of the respective isolates. The clinical and hematological alterations were evaluated by the determination of parasitaemia, haemogram, plasmatic fibrinogen,reticulocyte count, descriptive analysis of the bone marrow and erythrocytic osmotic fragility, for 30 days, totalizing seven moments of observation. The follow-up of the immunological response by the indirect fluorescent antibody test was carried out daily until the 10th day after inoculation (DAI) and after that, on the 151 20th, 25th and 30th DAI. A mild clinical manifestation of the disease was observed. The laboratory findings revealed low leveis of parasitaemia; decrease of the erythrogram values; absence of reticulocytes, initial decrease in the total count of leukocytes, neutrophils and lymphocytes with a posterior elevation of the number of these cells; hypercellularity of the erythrocytic series and decrease of the myeloid: erythroid relation which was more accentuated between the 8th and 12th DAI, and an increase of the erythrocytic osmotic fragility in the groups inoculated with the Southeast and Northeast isolates. None of the three isolates of B. bigemina gave rise to the clinical characteristic form of the disease, although they induced an humoral immune response.

• Experimental infection Babesia bigemina calves brazilian isolates haemogram bone marrow erythrocytic osmotic fragility Infecção experimental bezerros isolados hemograma medula óssea fragilidade osmótica eritrocitária.

Abstract in Portuguese:

RESUMO.- Mendonça C.L., Vieira D., Kohayagawa A., Schenk M.A.M., Madruga C.R. & Afonso J.A.B. 2003. [Clinical and hematological evaluation of Nelore calves experimentally infected with isolates of Babesia bigemina from the Southeastern, Northeastern and Northern regions of Brazil.] Avaliação clínica e hematológica em bezerros Nelore infectados experimentalmente com isolados de Babesia bigemina das regiões Sudeste, Nordeste e Norte do Brasil. Pesquisa Veterinária Brasileira 23(2):52-60. Clínica de Bovinos, Campus Garanhuns, Univ. Fed. Rural de Pernambuco, Cx. Postal 152, Garanhuns, PE 55292-901, Brazil. O presente trabalho teve por objetivo estudar comparativamente as alterações clínicas e hematológicas desencadeadas por isolados de Babesia bigemina das regiões Sudeste, Nordeste e Norte do Brasil em bezerros Nelore infectados experimentalmente. Foram utilizados 18 bezerros com idade entre sete e nove meses, isentos de anticorpos contra Babesia sp. e criados livres de carrapatos. Três animais foram previamente inoculados com 2,0x109 eritrócitos parasitados (EP) para cada isolado. Os outros 15 bezerros foram subdivididos em três grupos de cinco animais, que foram subinoculados com 1,0x1010 EP dos respectivos isolados. Foram avaliadas as alterações clínicas e hematológica por meio da determinação da parasitemia, do hemograma, do fibrinogênio plasmático, da contagem de reticulócitos, da análise descritiva da medula óssea e da fragilidade osmótica eritrocitária, no decorrer de 30 dias, perfazendo um total de sete momentos de observação. O acompanhamento da resposta imunológica pelo teste de imunofluorescência indireta foi realizado diariamente até o 10º dia pós-inoculação (DPI) e posteriormente no 15º, 20º, 25º e 30º DPI. Clinicamente, observou-se uma manifestação muito branda da doença. Os achados laboratoriais revelaram baixos níveis de parasitemia; decréscimo nos valores do eritrograma; ausência de reticulócitos; diminuição inicial na contagem total dos leucócitos, neutrófilos e linfócitos com posterior elevação do número destas células; hipercelularidade da série eritrocítica e decréscimo da relação mielóide:eritróide mais acentuada entre o 8º e 12º DPI e um aumento da fragilidade osmótica eritrocitária nos grupos inoculados com os isolados sudeste e nordeste. Nenhum dos três isolados de B. bigemina desencadeou a forma clínica característica da enfermidade, apesar de induzirem uma resposta imune humoral.

Abstract in English:

ABSTRACT.- Madruga C.R., Leal C.R.B., Ferreira A.M.T., Araújo F.R., Bonato A.L.V., Kessler R.H., Schenk M.A.M. & Soares C.O. 2002. Genetic and antigenic analysis of Babesia bigemina isolates from five geographical regions of Brazil. Pesquisa Veterinária Brasileira 22(4):153- 160. [Análise genética e antigênica de isolados de Babesia bigemina das cinco regiões fisiográficas do Brasil.] Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. A molecular epidemiological study was performed with Babesia bigemina isolates from five geographical regions of Brazil. The genetic analysis was done with random amplification of polymorphic DNA (RAPD), repetitive extragenic palindromic elements-polymerase chain reaction (REP-PCR) and enterobacterial repetitive intergenic consensus sequences-polymerase chain reaction (ERIC-PCR) that showed genetic polymorphism between these isolates and generated fingerprinting. In RAPD, IL0872 and IL0876 primers were able to detect at least one fingerprinting for each B. bigemina isolate. The amplification of B. bigemina DNA fragments by REP-PCR and ERIC-PCR gave evidence for the presence in this haemoprotozoan of the sequences described previously in microorganisms of the bacterial kingdom. For the first time it was demonstrated that both techniques can be used for genetic analysis of a protozoan parasite, although the ERIC-PCR was more discriminatory than REP-PCR. Toe dendogram with similarity coeficiente among isolates showed two clusters and one subcluster. The Northeastern and Mid-Westem isolates showed the greatest genetic diversity, while the Southeastem and Southem isolates were the closest. Toe antigenic analysis was done through indirect fluorescent antibodytechnique and Westem blotting using a panei of monoclonal antibodies directed against epitopes on the merozoite membrane surface, rhoptries and membrane of infected erythrocytes. As expected, the merozoite variable surface antigens, major surface antigen (MSA)-1 and MSA-2 showed antigenic diversity. However, B cell epitopes on rhoptries and infected erythrocytes were conserved among all isolates studied. In this study it was possible to identify variable and conserved antigens, which had already been described as potential immunogens. Considering that an attenuated Babesia clone used as immunogen selected populations capable of evading the immunity induced by this vaccine, it is necessary to evaluate more deeply the cross-protection conferred by genetically more distant Brazilian B. bigemina isolates and make an evaluation of the polymorphism degree of variable antigens such as MSA-1 and MSA-2.

• Babesia bigemina Brazilian isolates genetic polymorphism antigenic polymorphism isolados brasileiros polimorfismo genético polimorfismo antigênico.

Abstract in Portuguese:

RESUMO.- Madruga C.R., Leal C.R.B., Ferreira A.M.T., Araújo F.R., Bonato A.L.V., Kessler R.H., Schenk M.A.M. & Soares C.O. 2002. Genetic and antigenic analysis of Babesia bigemina isolates from five geographical regions of Brazil. Pesquisa Veterinária Brasileira 22(4):153- 160. [Análise genética e antigênica de isolados de Babesia bigemina das cinco regiões fisiográficas do Brasil.] Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. Um estudo de epidemiologia molecular foi executado com isolados de Babesia bigemina das cinco regiões fisiográficas do Brasil. A análise genética foi feita com amplificação aleatória de DNA polimórfico (RAPD), reação da polimerase em cadeia com seqüências de elementos extragênicos repetitivos palindrômicos (REP-PCR) e reação da polimerase em cadeia com seqüências repetitivas enterobacterianas intergênicas de consenso (ERIC-PCR) que apresentaram polimorfismo genético entre os isolados e geraram marcadores. No RAPD com os oligonucleotídeos iniciadores IL0872 e IL0876, foi possível detectar pelo menos um marcador por isolado de B. bigemina. A amplificação de fragmentos de DNA de B. bigemina por REPPCR e ERIC-PCR demonstrou a presença dessas seqüências, descritas anteriormente somente em microrganismos bacterianos, nesse hemoprotozoârio, e, pela primeira vez, foi verificado que podem ser utilizadas para análise genética de um protozoário. O ERIC-PCR foi mais discriminatório que o REP-PCR. O dendograma formado com o coeficiente de similaridade entre os isolados evidenciou dois agrupamentos e um subgrupo. Os isolados do Nordeste e Centro-Oeste demonstraram maior diversidade genética, enquanto que os isolados do Sudeste e Sul foram os mais próximos. A análise antigênica foi executada por meio de imunofluorescência indireta e Western blotting usando um painel de anticorpos monoclonais direcionados a epitopos B na membrana dos merozoítos, roptries e membrana de eritrócitos infectados. Os antígenos variáveis da superfície dos merozoítos, antígeno principal da superfície do merozoíto (APSM)-1 e APSM-2 apresentaram diversidade antigênica. Entretanto, os epítopos de células B nas roptries e nos eritrócitos infectados foram conservados em todos os isolados. Nesse estudo foi possível identificar antígenos variáveis e conservados que anteriormente haviam sido descritos como potenciais imunógenos. Considerando que um clone atenuado de Babesia utilizado para imunização selecionou populações capazes de evadir a resposta imune à vacina, torna-se necessário avaliar mais detalhadamente a imunidade cruzada existente entre os isolados brasileiros mais distantes geneticamente e realizar uma avaliação do grau de polimorfismo dos antígenos variáveis APSM-1 e APSM-2.

Abstract in English:

ABSTRACT.- Pagnani K.J.R., Pestana de Castro A.F., Gottschalk M., Silveira, W.D. & Nakazato G. 2002. [Serotyping of Streptococcus suis strains isolated from pigs in the States of São Paulo, Minas Gerais e Paraná, Brazil.] Sorotipagem de amostras de Streptococcus suis isolados de suínos em granjas dos Estados de São Paulo, Minas Gerais e Paraná. Pesquisa Veterinária Brasileira 22(1):1-5. Depto Microbiologia e Imunologia, Instituto de Biologia, Universidade Estadual de Campinas (Unicamp), Campinas, SP 13081-970, Brazil. Streptococcus suis infection in swine is common in all countries where hog production is well developed. This infection has been associated with bronchopneumonia, meningitis, arthritis, pericarditis, myocarditis, endocarditis, fibrinous polyserositis, septicaemia, rhinitis, and abortion. Streptococcus suis has also been described as a pathogen for ruminants and humans. In Brazil there are severa! clinical evidences about the existence of S. suis disease in pigs affecting more than 50% of farms in States of São Paulo, Minas Gerais and Paraná. In the present research 51 strains of S. suis isolated from piggeries of the States of São Paulo, Minas Gerais and Paraná were collected from different pathologies such as septicaemia, meningitis, arthritis and pneumonia and been recovered either in pure culture oras the predominant organism from porcine tissues. Culture of specimens was carried out on 5% bovine blood agar plates incubated at 37ºC for 24 hr: For the biochemical identification the a-hemolytic colonies of all capsulated isolates were submitted to various conventional tests, such as hydrolysis of arginine, Voges-Proskauer Test, and production of acid from various carbohydrates (inulin, salicin, trehalose, lactose, sucrose, sorbitol, mannitol and glycerol). The strains were also tested for their ability to grow in the presence of 6,5% NaCI and for the amylase production. In addition strains were tested by Api Strep 20 to confirm the identification of S.suis. For capsular typing only capsulated strains were typed by co-agglutination test, using antisera raised in rabbits against all reference strains from serotypes 1 to 8. Strains belonging to other serotypes were also typed. The co-agglutination was used for serotyping and the capsular reaction test was carried out for mieasuring the potency of the prepared antisera. From the total of 51 examined strains the following results were obtained, with regard to serotyping: 30 (58,8%) were serotype 2, 11 (21,6%) were serotype 3, seven (13, 72%) were serotype 7, two (3,92%) were serotype 1 and one strain belonged to serotype 14 (1,96%). As far as we are concerned, this is the first report on the isolation of a large number of S. suis strains in Brazil, from cases of illness caused by this bacterium among piglets. Also it was carried out serotyping of the isolates, showíng a high prevalence of serotype 2, as compared to other known serotypes of S. suis.

• Streptococcus suis isolation serotyping co-agglutination isolamento sorotipagem co-aglutinação.

Abstract in Portuguese:

RESUMO.- Pagnani K.J.R., Pestana de Castro A.F., Gottschalk M., Silveira, W.D. & Nakazato G. 2002. [Serotyping of Streptococcus suis strains isolated from pigs in the States of São Paulo, Minas Gerais e Paraná, Brazil.] Sorotipagem de amostras de Streptococcus suis isolados de suínos em granjas dos Estados de São Paulo, Minas Gerais e Paraná. Pesquisa Veterinária Brasileira 22(1):1-5. Depto Microbiologia e Imunologia, Instituto de Biologia, Universidade Estadual de Campinas (Unicamp), Campinas, SP 13081-970, Brazil. Infecções causadas por Streptococcus suis são muito comuns em países onde a indústria de carne suína é desenvolvida. Estas infecções estão relacionadas a casos clínicos de broncopneumonia, meningite, artrite, pericardite, miocardite, endocardite, poliserosite fibrinosa, septicemia, rinite e aborto. Esta bactéria também foi descrita como patógeno de ruminantes e humanos. No Brasil há evidências clínicas da existência de processos infecciosos causados por S. suis afetando mais de 50% das granjas em Estados como São Paulo, Minas Gerais e Paraná. No presente estudo foram isoladas 51 amostras de 5. suis de granjas do Estados acima referidos, coletadas de diferentes casos clínicos como septicemia, meningite, artrite e pneumonia, tendo sido obtidas ou em cultura pura ou como patógeno de maior predominância nos tecidos de suínos. Este material foi semeado em Columbia ágar sangue adicionado de 5% de sangue bovino e incubado a 37ºC por 24 horas. Para a identificação bioquímica as colônias que apresentavam a-hemólise, bem como as amostras padrão, foram submetidas a testes convencionais para a confirmação da espécie 5. suis, tais como: hidrólise de arginina, teste de Voges-Proskaue1: e produção de ácido a partir de vários carboidratos (inulina, salicina, trealose, lactose, sacarose, sorbitol, manitol e glicerol). As amostras também foram testadas para habilidade de crescimento em meio de TSA com 6,5% de NaCI e para a produção de amilase. Todas as amostras que fizeram parte desta pesquisa foram testadas pelo sistema Api 20 Strep para confirmação dos resultados obtidos nos testes convencionais. Para a sorotipagem foram produzidos antissoros de 1 a 8. Outras amostras não pertencentes a estes sorotipos também foram sorotipadas. O antissoro produzido em coelhos foi titulado pelo teste de aglutinação em tubo com 2-mercaptoetanol e pelo teste de reação capsular e, quando adequados, foram usados no teste de co-aglutinação, para a sorotipagem das amostras de 5. suis. A sorotipagem das 51 amostras isoladas mostraram os seguintes resultados: 30 (58,8%) foram classificadas corno sorotipo 2, 11 (21,6%) das amostras como sorotipo 3, sete (13,72%) como sorotipo 7, duas (3,92%) como sorotipo 1 e uma amostra como pertencente ao sorotipo 14 (1,96%). Este é o primeiro relato do isolamento de um grande número de amostras de 5. S. suis no Brasil, de casos típicos de processos infecciosos causados por esta bactéria. Também foi realizada a sorotipagem dos isolados, mostrando uma alta prevalência do sorotipo 2, quando comparada com a dos demais sorotipos encontrados.

Abstract in English:

ABSTRACT-. Reischak D., Ravazzolo A.P. & Moojen V. 2002. [lmmunofluorescence using Brazilian isolates for serological diagnosis of lentivirus infection in goats.] Imunofluorescência utilizando isolados brasileiros no diagnóstico soro lógico de infecção por lentivírus em caprinos. Pesquisa Veterinária Brasileira 22(1):7-12. Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: dilchak@hotmail.com Small ruminant lentiviruses (SRLV) are distributed worldwide and cause persistente infections in sheep and goats. The purpose of this work was to develop an indirect immunofluorescent antibody (IFA) test using Brazilian SRLV isolates for serological diagnosis of infection in goats. The IFA test was compared in its sensitivity and specificity to the AGID test in which Maedi-Visna WLC-1 strain was used as antigen. Ovine synovial secondary cell cultures infected with two goat SRLV isolates (CAEV UFRGS/Br-2 and CAEV UFRGS/Br-5) were used for the IFA. Goat serum samples (n = 239) were tested by the two tests. The AGID test detected antibodies in 129 (53.9%) serum samples. A higher number of animals was considered positive for the IFA. However, different results were obtained depending on the isolate used for the antigen preparation. When CAEV UFRGS/Br-2 was used as antigen, 216 (90.3%) sérum samples tested positive, against 213 (89.1%) with CAEV UFRGS/Br-5. There was no significant statistical difference between the antigens prepared with these two isolates. The IFA had sensitivity of 94.6% and 96.9%, and specificity of 14.5 % and 20%, when CAEV UFRGS/Br-2 and CAEV UFRGS/Br-5 were used as antigens, respectively. These results demonstrate that the IFA with antigens prepared with Brazilian SRLV isolates may play an important a role as a complementary serological test for the diagnosis of infections by these agents.

• Small ruminant lentiviruses Brazilian isolates diagnosis indirect immunofluorescence immunodiffusion goats Lentivírus de pequenos ruminantes isolados brasileiros diagnóstico imunofluorescência indireta imunodifusão em gel de ágar caprinos.

Abstract in Portuguese:

RESUMO.- Reischak D., Ravazzolo A.P. & Moojen V. 2002. [lmmunofluorescence using Brazilian isolates for serological diagnosis of lentivirus infection in goats.] Imunofluorescência utilizando isolados brasileiros no diagnóstico soro lógico de infecção por lentivírus em caprinos. Pesquisa Veterinária Brasileira 22(1):7-12. Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: dilchak@hotmail.com Os lentivírus de pequenos ruminantes (SRLV) têm distribuição mundial e causam infecções persistentes em ovinos e caprinos. O objetivo deste trabalho foi desenvolver um teste de imunofluorescência indireta (IFA), utilizando isolados brasileiros de SRLV, para o diagnóstico sorológico de infecção por estes agentes em caprinos. A técnica de IFA foi comparada, quanto à sensibilidade e à especificidade, ao teste de AGID com antígeno do vírus Maedi-Visna WLC-1. Cultivas celulares secundários de membrana sinovial ovina infectadas com dois isolados de SRLV de origem caprina (CAEV Br/UFRGS-2 e CAEV BiiUFRGS-5) foram utilizados para o teste de IFA. Duzentas e trinta e nove amostras de soro caprino foram submetidas aos dois testes. O teste de AGID detectou 129 (53.9%) amostras de soro caprino com anticorpos para SRLV. O teste de IFA detectou mais amostras reagentes, sendo que resultados diferentes foram observados de acordo com o isolado de SRLV empregado. Quando o isolado CAEV Br/UFRGS-2 foi utilizado corno antígeno, 216 (90.3%) amostras de soro caprino foram reagentes, enquanto que o isolado CAEVBr/UFRGS-5 detectou 213 (89.1%) amostras de soro positivas. Não houve diferença estatisticamente significativa entre esses dois isolados. O teste de IFA desenvolvido teve sensibilidade de 94.6% e 96.9% e especificidade 14.5% e 20%, quando os isolados CAEV Br/UFRGS-2 e CAEV Br/UFRGS-5 foram usados como antígeno, respectivamente. O aprimoramento da técnica, assim como sua comparação com um teste mais sensível, ainda se fazem necessários. No entanto, os resultados demonstraram que a técnica de IFA, utilizando isolados brasileiros de SRLV como antígeno, apresenta potencial como um teste alternativo e complementar para o diagnóstico sorológico de infecção por estes agentes.

Abstract in English:

ABSTRACT.- Hofer E., Silva Filho S.J. & Reis E.M.F. 1998. [Salmonella serovars isolated from feedstuff and poultry feeds in Brazil.] Sorovares de Salmonella isolados de matérias-primas e de ração para aves no Brasil. Pesquisa Veterinária Brasileira 18(1):21-27. Depto Bacteriologia, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ 21045-900, Brazil. Salmonella strains were isolated from feedstuff and poultry feeds from several regions of Brazil in 1976 and from 1979 to 1991. Serotyping of 2293 isolates showed 151 serovars which pertained to 17 serogroups and were classified as subspecies I (99.6%), III (0.33%) and IV (0.04%). There was a predominance of groups 0:7 (30.4%), 0:4 (24.5%), 0:3, 10 (19.1%), 0:13 (7 .8%), 0:1, 3, 19 (4.9%) and 0:18 (3. 7%), representing 90% of the serogroups characterized that accounted for 103 different serotypes (68.2%). Predominant serovars isolated from all sources were S. Montevideo, S. Senftenberg, S. Havana, S. Mbandaka, S. Tennessee, S. Infantis, S. Agona, S. Anatum, S. Cerro and S. Bredeney. Bacteriological and epidemiological aspects and the relationship with serovars isolated from poultry are discussed.

• Poultry feeds Salmonella serogroup serovar prevalence Matéria-prima ração aves sorogrupos sorovares.

Abstract in Portuguese:

RESUMO.- Hofer E., Silva Filho S.J. & Reis E.M.F. 1998. [Salmonella serovars isolated from feedstuff and poultry feeds in Brazil.] Sorovares de Salmonella isolados de matérias-primas e de ração para aves no Brasil. Pesquisa Veterinária Brasileira 18(1):21-27. Depto Bacteriologia, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ 21045-900, Brazil. Foram caracterizadas antigenicamente amostras de Salmonella isoladas de matérias-primas e de ração para aves em 1976 e durante doze anos consecutivos (1979-1991). As 2293 culturas analisadas provieram de sete regiões distintas do país e possibilitaram o reconhecimento de 151 sorovares, classificados bioquimicamente nas subespécies I (99,6%) IIIa (0,33%) e IV (0,04%), respectivamente. Os sorovares identificados se distribuiram por 17 sorogrupos, com predominância de 0:7 (30,4%), 0:4 (24,5%), 0:3, 10 (19, 1%), 0:13 (7,8%), 0:1,3,19 (4,9%) e 0:18 (3,7), que representam 90% dos grupos sorológicos caracterizados e constituídos de 103 (68,2%) sorotipos. Dentre os dez sorovares mais frequentemente reconhecidos citam-se S. Montevideo, S. Senftenberg, S. Havana, S. Mbandaka, S. Tennessee, S. Infantis, S. Agona, S. Anatum, S. Cerro e S. Bredeney. Alguns aspectos de caráter epidemio-lógico foram discutidos, envolvendo particularmente, determinados sorotipos e·inclusive confrontando-se os resultados obtidos com aqueles oriundos de investigação conexa em aves.