PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

Assis R.A., Lobato F.C.F., Serakides R., Santos R.L., Dias G.R.C., Nascimento R.A.P., Abreu V.L.V, Parreiras P.M. & Uzal F.A. 2005. Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs. Pesquisa Veterinária Brasileira 25(1):4-8. Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Presidente Antônio Carlos 6627, Cx. Postal 567, Belo Horizonte, MG 30123-970, Brazil. E-mail: assisra@rwnet.com.br Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

• Blackleg malignant oedema Clostridia guinea pigs fluorescent antibody technique streptavidin biotin peroxidase technique.

Abstract in Portuguese:

Assis R.A., Lobato F.C.F., Serakides R., Santos R.L., Dias G.R.C., Nascimento R.A.P., Abreu V.L.V, Parreiras P.M. & Uzal F.A. 2005. Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs. Pesquisa Veterinária Brasileira 25(1):4-8. Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Presidente Antônio Carlos 6627, Cx. Postal 567, Belo Horizonte, MG 30123-970, Brazil. E-mail: assisra@rwnet.com.br Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

Abstract in English:

Minho A.P., Navarro I.T., Freire R.L., Vidotto O., Gennari S.M., Marana E.M. & Garcia J.L. 2004. Evaluation of the indirect fluorescent antibody test and modified agglutination test for detection of antibodies against Toxoplasma gondii in experimentally infected pigs. Pesquisa Veterinária Brasileira 24(4):199-202. Depto Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Cx. Postal 6001, Londrina, PR 86050-970, Brazil. E-mail: italmar@uel.br The study determined the sensitivity and specificity of the indirect fluorescent antibody test (IFAT) and modified agglutination test (MAT) for anti-Toxoplasma gondii antibody detection by analyzing sera from 46 experimentally infected pigs. Values for sensitivity were 95.7% (confidence interval 95%: 84.0-99.2%) and for specificity 97.8% (confidence interval 95%: 87.0-99.9%) in both tests. There was an optimum agreement of results between IFAT and MAT evidenced by a Kappa test of 0.86. These results validate these tests for the detection of T. gondii infection in pigs. IFAT and MAT despite methodologies with different characteristics and readings have similar accuracy in pig serum samples.

• Toxoplasma gondii swine antibodies serology indirect fluorescent antibody test (IFAT) modified agglutination test (MAT).

Abstract in Portuguese:

Minho A.P., Navarro I.T., Freire R.L., Vidotto O., Gennari S.M., Marana E.M. & Garcia J.L. 2004. Evaluation of the indirect fluorescent antibody test and modified agglutination test for detection of antibodies against Toxoplasma gondii in experimentally infected pigs. Pesquisa Veterinária Brasileira 24(4):199-202. Depto Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Cx. Postal 6001, Londrina, PR 86050-970, Brazil. E-mail: italmar@uel.br The study determined the sensitivity and specificity of the indirect fluorescent antibody test (IFAT) and modified agglutination test (MAT) for anti-Toxoplasma gondii antibody detection by analyzing sera from 46 experimentally infected pigs. Values for sensitivity were 95.7% (confidence interval 95%: 84.0-99.2%) and for specificity 97.8% (confidence interval 95%: 87.0-99.9%) in both tests. There was an optimum agreement of results between IFAT and MAT evidenced by a Kappa test of 0.86. These results validate these tests for the detection of T. gondii infection in pigs. IFAT and MAT despite methodologies with different characteristics and readings have similar accuracy in pig serum samples.

Abstract in English:

Rossetti C.A., Vansco B.N., Pini, N & Carfagnini J.C. 2004. Comparison of three diagnostic techniques for the detection of leptospires in the kidneys of wild house mice (Mus musculus). Pesquisa Veterinária Brasileira 24(1):6-10. Instituto de Patobiología, Centro Nacional de Investigaciones Agropecuarias (CNIA) del Instituto Nacional de Tecnología Agropecuaria (INTA), CC 25 (1712) Castelar, Buenos Aires, Argentina. E-mail: crossetti@cicv.inta.gov.ar Forty-one wild house mice (Mus musculus) were trapped in an urban area, near railways, in Santa Fe city, Argentina. Both kidneys from each mouse were removed for bacteriological and histological examination. One kidney was inoculated into Fletcher semi-solid medium and isolates were serologically typed. The other kidney was microscopically examined after hematoxylin-eosin, silver impregnation and immunohistochemical stains. Leptospires, all of them belonging to the Ballum serogroup, were isolated from 16 (39%) out of 41 samples. The presence of the agent was recorded in 18 (44%) and in 19 (46%) out of 41 silver impregnated and immunohistochemically stained samples respectively. Additionally, leptospires were detected in high number on the apical surface of epithelial cells and in the lumen of medullary tubules and they were less frequently seen on the apical surface of epithelial cells or in the lumen of the cortical tubules, which represents an unusual finding in carrier animals. Microscopic lesions consisting of focal mononuclear interstitial nephritis, glomerular shrinkage and desquamation of tubular epithelial cells were observed in 13 of 19 infected and in 10 of 22 non-infected mice; differences in presence of lesions between infected and non-infected animals were not statistically significant (P=0,14). The three techniques, culture, silver impregnation and immunohistochemistry, had a high agreement (k³0.85) and no significant differences between them were detected (P>0.05). In addition, an unusual location of leptospires in kidneys of carrier animals was reported, but a relationship between lesions and presence of leptospires could not be established.

• Leptospires diagnostic techniques Mus musculus lesions.

Abstract in Portuguese:

Rossetti C.A., Vansco B.N., Pini, N & Carfagnini J.C. 2004. Comparison of three diagnostic techniques for the detection of leptospires in the kidneys of wild house mice (Mus musculus). Pesquisa Veterinária Brasileira 24(1):6-10. Instituto de Patobiología, Centro Nacional de Investigaciones Agropecuarias (CNIA) del Instituto Nacional de Tecnología Agropecuaria (INTA), CC 25 (1712) Castelar, Buenos Aires, Argentina. E-mail: crossetti@cicv.inta.gov.ar Forty-one wild house mice (Mus musculus) were trapped in an urban area, near railways, in Santa Fe city, Argentina. Both kidneys from each mouse were removed for bacteriological and histological examination. One kidney was inoculated into Fletcher semi-solid medium and isolates were serologically typed. The other kidney was microscopically examined after hematoxylin-eosin, silver impregnation and immunohistochemical stains. Leptospires, all of them belonging to the Ballum serogroup, were isolated from 16 (39%) out of 41 samples. The presence of the agent was recorded in 18 (44%) and in 19 (46%) out of 41 silver impregnated and immunohistochemically stained samples respectively. Additionally, leptospires were detected in high number on the apical surface of epithelial cells and in the lumen of medullary tubules and they were less frequently seen on the apical surface of epithelial cells or in the lumen of the cortical tubules, which represents an unusual finding in carrier animals. Microscopic lesions consisting of focal mononuclear interstitial nephritis, glomerular shrinkage and desquamation of tubular epithelial cells were observed in 13 of 19 infected and in 10 of 22 non-infected mice; differences in presence of lesions between infected and non-infected animals were not statistically significant (P=0,14). The three techniques, culture, silver impregnation and immunohistochemistry, had a high agreement (k³0.85) and no significant differences between them were detected (P>0.05). In addition, an unusual location of leptospires in kidneys of carrier animals was reported, but a relationship between lesions and presence of leptospires could not be established.

Abstract in English:

ABSTRACT.- Scherer C.F.C., Flores E.F., Weiblen R., Kreutz L.C., Dürr J.W., Brum L.P., Quadros V.L. & Lima M. 2000. [A rapid virus-neutralization test for detection of antibodies against bovine viral diarrhea virus (BVDV) in milk.] Técnica rápida de neutralização viral para pesquisa de anticorpos contra o vírus da Diarréia Viral Bovina (BVDV) no leite. Pesquisa Veterinária Brasileira 22(2):45-50. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. The identification of bovine viral diarrhea virus (BVDV) positive herds through detection of antibodies in milk may viabilize large scale control/eradication programs. With this objective, the virus neutralization test (VN) was adapted to detect BVDV antibodies in milk. The adaptation consisted of a reduction in the time of incubation followed by detection of viral antigens in the indicator cells by immunofluorescence (IFA) and allowed readings at 24 hours. The rapid virus neutralization test (RVN) was initially tested in 1,335 serum samples, showing a 93. 7% sensitivity and 91.1 % agreement with the traditional VN. The RVN was also used to test 423 bovine sera that were toxic for cell culture in the traditional VN test, detecting 316 (74.7%) positive samples. Testing of matched serum and milk samples from BVDV seropositive cows showed that the VNR can detect antibodies in the milk of cows with serum neutralizing titers as low as 10. Anti-BVDV neutralizing activity was detected in milk of 97.4% (191/196) of cows with serum titers 3320; in 92.9% (79/85) of cows with titers of 160; in 88% (59/67) of cows with serum titers of 80. The frequency of BVDV antibodies in milk was 76.9% (40/52) for cows with serum titers of 40; 61.3% (19/31) for cows with titers of 20 and 33.3% (10/30) for cows with serum titers of 20. These results demonstrate that the RVN test is adequate for detecting BVDV antibodies in milk, mainly in cows having moderate to high serum titers, and therefore may be used for testing bulk milk samples to identify herds with viral activity. The use of this test may viabilize large scale programs for control/eradication of BVDV infection. It allows to assay a large number of samples and identify positive herds through testing milk routinely submitted for somatic cell counts (SCC), reducing costs with individual sample collection, shipping and testing.

• Bovine viral diarrhea virus BVDV diagnosis antibodies milk Vírus da Diarréia Viral Bovina diagnóstico leite anticorpos.

Abstract in Portuguese:

RESUMO.- Scherer C.F.C., Flores E.F., Weiblen R., Kreutz L.C., Dürr J.W., Brum L.P., Quadros V.L. & Lima M. 2000. [A rapid virus-neutralization test for detection of antibodies against bovine viral diarrhea virus (BVDV) in milk.] Técnica rápida de neutralização viral para pesquisa de anticorpos contra o vírus da Diarréia Viral Bovina (BVDV) no leite. Pesquisa Veterinária Brasileira 22(2):45-50. Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil. A identificação de rebanhos positivos para o vírus da Diarréia Viral Bovina (BVDV) através de detecção de anticorpos no leite pode viabilizar programas de controle em larga escala. Com esse objetivo, a técnica de soroneutralização (SN) foi adaptada para a pesquisa de anticorpos em amostras de leite. A adaptação consistiu na redução do tempo de incubação do teste, seguida da detecção de antígenos virais por imunofluorescência. A redução do tempo de incubação minimizou os efeitos tóxicos do leite sobre as células de cultivo, além de permitir a obtenção dos resultados em 24 horas. A técnica rápida (SNR) foi inicialmente testada em 1.335 amostras de soro bovino, apresentando sensibilidade de 93,7% e concordância de 91, 1% em relação à SN tradicional. A SNR foi também utilizada para testar 423 amostras de soro bovino que apresentaram toxicidade para as células na SN tradicional, detectando 316 (74,7%) amostras positivas. O teste de amostras de soro e leite de 520 vacas em lactação demonstrou que a SNR pode detectar anticorpos no leite de vacas com títulos séricos a partir de 10. Atividade neutralizante anti-BVDV no leite foi detectada em 97,4% (191/196) de vacas com títulos séricos 3 320; em 92,9% (79/85) de vacas com títulos de 160; em 88% (59/67) de vacas títulos de 80. A freqüência de animais positivos na SNR foi de 76,9% (40/52) para animais com títulos séricos de 40; 61,3% (19/31) com títulos de 20 e de 33,3% (10/30) para vacas com títulos de 10. Esses resultados demonstram que a técnica de SNR é adequada para a pesquisa de anticorpos anti-BVDV no leite, principalmente em animais com títulos moderados e altos de anticorpos. Essa técnica pode ser utilizada para testar amostras coletivas de leite e identificar rebanhos com atividade viral. A utilização dessa técnica pode viabilizar programas regionais de combate à infecção, pois permite testar um grande número de amostras e identificar rebanhos positivos através do leite enviado rotineiramente para contagem de células somáticas (CCS), reduzindo significativamente os custos com a coleta individual, transporte e teste de amostras.

Abstract in English:

ABSTRACT.- Madruga C.R., Marques A.P.C., Araújo F.R., Miguita M., Carvalho C.M.E., Araújo F.S., Umaki A.C.S., Crocci A.J. & Queiróz R.A. 2001. Evaluation of an ELISA for detection of antibodies to Babesia bigemina in cattle and it's application in an epidemiological survey in Brazil. [Avaliação de um ELISA para detecção de anticorpos contra Babesia bigemina em bovinos e sua aplicação em um inquérito sorológico no Brasil.] Pesquisa Veterinária Brasileira 21 (2):72-76. Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. An indirect enzyme-linked immunosorbent assay (ELISA) using a crude antigen was evaluated for its performance to detect Babesia bigemina antibodies. The sensitivity and specificity were 98.0% and 99.0%, respectively. ln agreement with the high specificity, no cross-reactions were verified with sera from calves inoculated three times with 107 Babesia bovis organisms. With regard to the comparison of ELISA and indirect fluorescent antibody test (lFAT) in detecting antibodies against B. bigemina in calves experimentally infected with five Brazilian geographical isolates of this hemoparasite, lFAT was able to detect antibodies one day earlier in most of the calves' sera. There was a good agreement between results shown by ELISA and IFAT with sera from an enzootically stable area (k=0.61). However, there was no agreement between these serological tests with sera from an enzootically unstable area (k=0.33). The ELISA was employed in an epidemiological survey using with 1,367 sera from four counties in the Pantanal of Mato Grosso do Sul and characterized this region as an enzootically stable area, since the prevalence ranged from 87.7 to 98.9%. Therefore, this ELISA with high sensitivity, specificity and performance similar to lFAT can be employed in serological diagnosis of B. bigemina.

• Babesia bigemina ELISA serological tests cattle Pantanal testes sorológicos bovinos Pantanal.

Abstract in Portuguese:

RESUMO.- Madruga C.R., Marques A.P.C., Araújo F.R., Miguita M., Carvalho C.M.E., Araújo F.S., Umaki A.C.S., Crocci A.J. & Queiróz R.A. 2001. Evaluation of an ELISA for detection of antibodies to Babesia bigemina in cattle and it's application in an epidemiological survey in Brazil. [Avaliação de um ELISA para detecção de anticorpos contra Babesia bigemina em bovinos e sua aplicação em um inquérito sorológico no Brasil.] Pesquisa Veterinária Brasileira 21 (2):72-76. Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. Um ensaio de imunoadsorção enzimática (ELISA) baseado em antígeno bruto foi avaliado na detecção de anticorpos contra Babesia bigemina. A sensibilidade e a especificidade do teste foram de 98,0% e 99,0%, respectivamente. Concordando com a alta especificidade do teste, não foram verificadas reações cruzadas com soros de bezerros inoculados três vezes com 107 merozoítos de Babesia bovis. Com relação à comparação do ELISA com a imunofluorescência indireta (IFAT) na detecção de anticorpos contra B. bigemina em bezerros experimentalmente infectados com cinco isolados brasileiros geograficamente distintos deste hemoparasito, o IFAT foi capaz de detectar anticorpos um dia antes do ELISA na maioria dos soros dos animais. Houve urna boa concordância entre os resultados encontrados no ELISA e no IFAT com soros de bovinos de região de estabilidade enzoótica (k=0.61 ). No entanto, não houve concordância entre os testes sorológicos com soros de animais de área de instabilidade enzoótica (k=0.33). O ELISA foi empregado em um inquérito epidemiológico com 1.367 soros de quatro municípios do Pantanal de Mato Grosso do Sul e caracterizou esta região corno urna área de estabilidade enzoótica, urna vez que as prevalências variaram de 87, 7 a 98,9%. Dessa forma, este ELISA, que apresentou alta sensibilidade, especificidade e desempenho similar ao IFAT, pode ser utilizado no diagnóstico sorológico de B. bigemina.

Abstract in English:

ABSTRACT.- Madruga C.R., Kessler R.H., Schenk M.A.M. & Miguita M. 2000. A conglutination test for a rapid detection of antibodies against Babesia bigemina. [Teste de conglutinação rápida para detecção de anticorpos contra Babesia bigemina.] Pesquisa Veterinária Brasileira 20(4):161-166. Embrapa Gado de Corte, Rodovia BR 262, Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. A rapid conglutination test (RCT) with performance comparable to the indirect fluorescent antibody technique (IFAT) was developed to detect antibodies against Babesia bigemina (B. bigemina-RCT). The B. bigemina-RCT is a sensitive, specific, economical, and rapidly performed serological test suitable for field application or minimally equipped laboratories. This test had a sensitivity of 90.9%, and specificity of 97.6%, compared to IFAT, which showed for the sarne parameters respectively, 98.3% and 99.7%. The early detection of anti- B. bigemina immunoglobulins by RCT in experimental infections was nearly parallel to that of IFAT. Cross reactions were observed with sera from calves experimentally infected with Babesia bovis (1.8%) and with Anaplasma marginale (1.2%). RCT antigen prepared with non parasitized erythrocytes (negative antigen) showed 1.5%, 3.5% and 2.2% of positive reactions with sera from animals experimentally infected with B. bigemina, B. bovis and A. marginale. However, none of the sera from animals of endemic areas for babesia infection resulted in positive reactions with the negative antigen. Considering these results and shelf life over six months, the B. bigemina-RCT could be used for epidemiological surveys and evaluation of control measures against this species of Babesia.

• Babesia bigemina antibodies serological test conglutination anticorpos teste sorológico conglutinação.

Abstract in Portuguese:

RESUMO.- Madruga C.R., Kessler R.H., Schenk M.A.M. & Miguita M. 2000. A conglutination test for a rapid detection of antibodies against Babesia bigemina. [Teste de conglutinação rápida para detecção de anticorpos contra Babesia bigemina.] Pesquisa Veterinária Brasileira 20(4):161-166. Embrapa Gado de Corte, Rodovia BR 262, Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. Um teste rápido de conglutinação (TCR) com desempenho comparável a imunofloorencência indireta (IFI) foi desenvolvido para detectar anticorpos contra Babesia bigemina. O TCR:-B.bigemina é um teste sorológico sensível, econômico e executável rapidamente; apropriado para condições de campo ou laboratórios com estrutura mínima. Este teste tem uma sensibilidade de 90,9% e especificidade de 97,6%, enquanto que a IFI apresentou para os mesmos parâmetros, respectivamente, 98,3% e 99,7%. Nas infecções experimentais a detecção de imunoglobulinas anti-B. bigemina pelo TCR foi aproximadamente a mesma da IFI. As reações cruzadas verificadas nos soros de bezerros experimentalmente infectados com Babesia bovis e Anaplasma marginale foram 1,8% e 1,2%, respectivamente. O antígeno preparado com eritrócitos não parasitados (antígeno negativo) apresentou 1,5%, 3,5% e 2,2% de reações positivas com os soros de animais infectados corri ·B. bigemina, B. bovis e A. marginale. Entretanto, nenhum dos soros dos animais de áreas endêmicas para infecção de babésia resultaram ern reações positivas com o antígeno negativo. Consideraodo,estes resultados e o período de viabilidade do antígeno de TCR, acima de seis meses, possibilita o TCR-B. bigemina ser utilizado em levantamentos epidemiológicos e na avaliação das medidas de controle contra esta espécie de Babesia.

Abstract in English:

ABSTRACT.- Madruga C.R., Araújo F.R., Marques A.P.C., Carvalho C.M.E., Cusinato F.Q., Crocci A.J., Kessler R.H. & Miguita M. 2000. [Development of an enzyme-linked immunosorbent assay for the detection of antibodies against Babesia bovis.] Desenvolvimento de uma prova de imunoadsorção enzimática para detecção de anticorpos contra Babesia bovis. Pesquisa Veterinária Brasileira 20(4):167-170. Embrapa Gado de Corte, BR 262 Km 4, Campo Grande, MS 79002-970, Brazil. An enzyme-linked immunosorbent assay (ELISA) for antibodies to Babesia bovis was developed and evaluated in comparison with the indirect fluorescent antibody test (IFAT). The ELISA sensitivity and specificity, estimated with 100 positive sera from cattle experimentally infected with B. bovis and 108 negative sera collected from B. bovis-free herds, were 98.0% and 98.1 %, respectively. Positive and negative predictive values were, respectively, 98.0% and 98.1 %, and precision was 98.1 %. No cross-reactions were detected with 80 sera from calves experimentally inoculated with Babesia bigemina. The ELISA was compared with IFAT using 110 cattle sera from an enzootically stable area and with 168 cattle sera from an enzootically unstable area. In both cases, there was a significant agreement between results of both tests (P=0.631 and 0.4725, respectively). In an epidemiological study performed with ELISA in the Pantanal region of the State of Mato Grosso do Sul with 1,365 cattle sera, 83.9%were positive for antibodies against B. bovis, characterizing this region as enzootically stable.

• Babesia bovis ELISA indirect fluorescent antibody test cattle epidemiological survey Pantanal Mato Grosso do Sul Brazil imunofluorescência indireta bovino levantamento sorológico Pantanal Mato Grosso do Sul Brasil.

Abstract in Portuguese:

RESUMO.- Madruga C.R., Araújo F.R., Marques A.P.C., Carvalho C.M.E., Cusinato F.Q., Crocci A.J., Kessler R.H. & Miguita M. 2000. [Development of an enzyme-linked immunosorbent assay for the detection of antibodies against Babesia bovis.] Desenvolvimento de uma prova de imunoadsorção enzimática para detecção de anticorpos contra Babesia bovis. Pesquisa Veterinária Brasileira 20(4):167-170. Embrapa Gado de Corte, BR 262 Km 4, Campo Grande, MS 79002-970, Brazil. Uma prova de imunoadsorção enzimática (ELISA) para detecção de anticorpos contra Babesia bovis foi desenvolvida e avaliada em comparação à imunofluorescência indireta (IFI). A sensibilidade e especificidade do ELISA, determinadas pela análise de 100 soros positivos de bovinos infectados experimentalmente com B. bovis e 108 soros negativos colhidos de bovinos livres de infecção por este hemoparasito, foram de 98,0% e 98, 1 %, respectivamente. Os valores preditivos positivo e negativo foram, respectivamente, 98,0% e 98, 1 % e a precisão do teste foi de 9.8, 1 %. Não foram detectadas reações cruzadas com 80 soros de bezerros experimentalmente inoculados com Babesia bigemina. O ELISA foi comparado à IFI usando 110 soros de rebanhos de área de estabilidade endêmica e 168 soros de rebanhos de áreas de instabilidade endêmica. Em ambos os casos, houve concordância significativa (P=0,631 e 0,4725, respectivamente) entre os resultados demonstrados pelos dois testes. Em um estudo epidemiológico realizado com o ELISA na região do Pantanal de Mato Grosso do Sul, com 1.365 soros de bovinos, 83,9% foram positivos para anticorpos contra B. bovis, caracterizando a região estudada como endemicamente estável.

Abstract in English:

ABSTRACT.- Caldas A.P.F., Leal E.S., Silva E.F.A. & Ravazzolo A.P. [Detection of feline immunodeficiency provirus in domestic cats by polymerase chain reaction.] Detecção do provírus da Imunodeficiência Felina em gatos domésticos pela técnica de Reação em Cadeia da Polimerase. Pesquisa Veterinária Brasileira 20(1):20-25. Centro de Biotecnologia/Faculdade de Veterinária, UFRGS, Av. Bento Gonçalves 9500, Porto Alegre, RS 91501-970, Brazil. Feline immunodeficiencyvirus (FIV) infection of domestic cats is one of the most promising animal models for the infection by the human immunodeficiency virus (HIV) which causes acquired immunodeficiency syndrome (AIDS). lnfected cats may develop a disease similar to that observed in AIDS patients, with increased susceptibility to opportunistic infections. Ln this study we used the polymerase chain reaction (PCR) to detect proviral DNA of feline immunodeficiency virus on the blood and tissue samples from cats with a clinical diagnosis of immunodeficiency. The PCR primers were used to amplify the gag gene, which is conserved among different isolates. From 40 samples analyzed, 15 were positive and 4 of them were submitted to hybridization to confirm the specificity of the amplified fragments. These results confirm the presence of FIV in domestic cats in Rio Grande do Sul, Brazil.

• Feline immunodeficiency virus FIV polymerase chain reaction PCR Vírus da Imunodeficiência Felina Reação em Cadeia da Polimerase.

Abstract in Portuguese:

SINOPSE.- Caldas A.P.F., Leal E.S., Silva E.F.A. & Ravazzolo A.P. [Detection of feline immunodeficiency provirus in domestic cats by polymerase chain reaction.] Detecção do provírus da Imunodeficiência Felina em gatos domésticos pela técnica de Reação em Cadeia da Polimerase. Pesquisa Veterinária Brasileira 20(1):20-25. Centro de Biotecnologia/Faculdade de Veterinária, UFRGS, Av. Bento Gonçalves 9500, Porto Alegre, RS 91501-970, Brazil. A infecção de gatos domésticos pelo Vírus da Imunodeficiência Felina (FIV) é um dos modelos mais promissores para o estudo da infecção pelo vírus da imunodeficiência humana (HIV) que causa a Síndrome de Imunodeficiência Adquirida (AIDS). O FIV causa, em gatos, uma enfermidade similar àquela observada em pacientes com AIDS, sobretudo no que diz respeito ao aumento da susceptibilidade a infecções oportunistas. No presente estudo, utilizou-se a Reação em Cadeia da Polimerase (PCR), com o objetivo de detectar o provírus do FIV em gatos com sinais clínicos de imunodeficiência. O fragmento de DNA escolhido como alvo para amplificação situa-se no gene gag do lentivírus felino, o qual é conservado entre as diferentes amostras do vírus. O DNA utilizado foi extraído a partir de amostras de sangue e de tecidos de animais com suspeita clínica de imunodeficiência. Das 40 amostras analisadas, 15 foram positivas, das quais 4 foram submetidas à hibridização, confirmando a especificidade dos fragmentos amplificados. Esses resultados demonstram a presença do FIV na população de gatos domésticos do Rio Grande do Sul, Brasil.

Abstract in English:

ABSTRACT.- Monteavaro, C.E., Soto P., Echevarría H.M., Catena M.C., Portiansky E.L. & Gimeno E.J. 2000. Immunohistochemical detection of Tritrichomonas foetus in experimentally infected mice. [Detecção imunohistoquímica de Tritrichomonas foetus em camundongos experimentalmente infectados.] Pesquisa Veterinária Brasileira 20(1):43-46. Institute of Pathology, Veterinary School, UNLP, P.O.Box 296, 1900 La Plata, Argentina. The need to intensify knowledge of the pathogenesis of bovine genital trichomoniasis (BGT) led to the use ofalternative animal models such as the mouse. Nevertheless, it is necessary to elucidate the dynamics of the infection in this animal species, evaluating different stages of the colonization and evolution of the pathological alterations. The immunohistochemistry (IHC) of fers advantages over the routine histopathological staining techniques for the detection of the protozoan in tissues, cellular detritus and inside the macrophages. The goal of the present study was to demonstrate the presence of Tritrichomonas foetus in the reproductive tract of infected mice using an IHC technique. Female BALB/c mice were infected with a suspension of T. foetus by intravaginal route, in the estrum phase, detected by exfoliative vaginal cytology. After 10 weeks, the animais were sacrificed; uterus and vagina were fixed and histologically processed. Some slides were stained with HE. The rest of the slides were processed for IHC. An immunoadsorbed polyclonal serum against T. foetus was used. The. avidine-biotine technique (HistoMouse, Zymed ™) was employed. The histopathological studies showed a dilation of the uterine glands, presence of macrophages in the lumen of the organ and inner part of the endometrial glands. No T. foetus was identified tísing this method. The IHQ allowed additionally the identification of the protozoan in the endometrium, endometrial glands, uterine lumen and inside neutrophils and macrophages. The cytological studies stained with IHC showed either isolated T. foetus adhered to epithelial cells or inside macrophages. This technique proves to be a useful tool for the study of the pathogenesis of bovine genital trichomoniasis (BGT) in an experimental model.

• Tritrichomonas foetus bovine genital trichomoniasis immunohistochemistry mouse tricomoníase genital bovina imunohistoquímica camundongo.

Abstract in Portuguese:

RESUMO.- Monteavaro, C.E., Soto P., Echevarría H.M., Catena M.C., Portiansky E.L. & Gimeno E.J. 2000. Immunohistochemical detection of Tritrichomonas foetus in experimentally infected mice. [Detecção imunohistoquímica de Tritrichomonas foetus em camundongos experimentalmente infectados.] Pesquisa Veterinária Brasileira 20(1):43-46. Institute of Pathology, Veterinary School, UNLP, P.O.Box 296, 1900 La Plata, Argentina. A necessidade de aumentar o conhecimento da patogenia da tricomoníase genital bovina (BGT) conduziu ao uso de modelos experimentais alternativos como o camundongo. Não obstante, é necessário elucidar a dinâmica da infecção nesta espécie e avaliar as diferentes fases da colonização e evolução das alterações patológicas. A imunohistoquí-mica (IHQ) oferece vantagens sobre as técnicas histoquímicas de rotina para a observação do protozoário em tecidos, detritos celulares e dentro de macrófagos. O objetivo do presente trabalho foi demonstrar pelo uso de uma técnica de IHQ a presença de Tritrichomonas foetus no sistema reprodutivo de camundongos infectados. Camundongos BALB/c fêmeas foram infectados pela via intravaginal, com uma suspensão de T. foetus, na fase de estro, detectado com citologia exfoliativa vaginal. Depois de 10 semanas, os animais foram sacrificados; útero e vagina forma fixados e processados para histologia. Alguns cortes foram corados com HE. O restante dos cortes foi processado para IHQ. Foi usado um soro policlonaf imunoadsorvído anti-T.foetus. A técnica de avidina biatina (HistoMouse, Zymed™) foi empregada. Os estudos histopatológicos mostraram uma dilatação das glândulas uterinas, presença de macrófagos no lúmen do órgão e parte interna das glândulas endometriais. T. foetus não foi identificado por esse método. A IHQ permitiu identificar as mesmas lesões observadas e a presença do protozoário no endométrio, nas glândulas endometriais, no lúmen uterino e dentro de neutrófilos e macrófagos. O estudo citológico em lâminas coradas por IHQ, mostrou T.foetus aderido a células epiteliais, ou dentro de macrófagos. Esta técnica demonstra ser uma ferramenta útil para o estudo da patogenia da tricomoníase genital bovina (BGT) utilizando-se o camundongo como modelo experimental.

Abstract in English:

ABSTRACT.- Fernández P.E., Sanguinetti H.R., Portiansky E.L, Barbeito C.G. & Gimeno E.J. 1999. Detection of illegal estrogen administration through immunohistochemical markers in the bovine prostate. [Verificação da administração ilegal de estrógenos através de marcadores imunohistoquímicos na próstata de bovinos] Pesquisa Veterinaria Brasileira 19(3/4):133-138. Institute of Pathology "Prof. Dr. Bernardo Epstein", School of Veterinary Sciences, National University of La Plata, P.O.Box 296, (1900) La Plata, Argentina. The immunodetection of diverse cell markers was evaluated in prostatic samples from bullocks, and bullocks showing epithelial hyperplasia-metaplasia, with oestrogen-induced changes, and in experimental samples from bullocks inoculated with dietylstilbestrol (DES). Antigen-retrieval procedures allowed the use of tissues that had been fixed in formalin for long periods. Three tissue markers were chosen for the study: cytokeratins 13 and 16, vimentin and desmin. Monoclonal antibody K8.12 (specific for cytokeratins 13 and 16) stained basal cells and hyperplastic-metaplastic epithelium; monoclonal antivimentin, and desmin, allowed the definition of fibromuscular changes.

• Bovine prostate cell markers immunohistochemistry oestrogen Próstata bovina marcadores de celulares imunohistoquímica estrógenos.

Abstract in Portuguese:

RESUMO.- Fernández P.E., Sanguinetti H.R., Portiansky E.L, Barbeito C.G. & Gimeno E.J. 1999. Detection of illegal estrogen administration through immunohistochemical markers in the bovine prostate. [Verificação da administração ilegal de estrógenos através de marcadores imunohistoquímicos na próstata de bovinos] Pesquisa Veterinaria Brasileira 19(3/4):133-138. Institute of Pathology "Prof. Dr. Bernardo Epstein", School of Veterinary Sciences, National University of La Plata, P.O.Box 296, (1900) La Plata, Argentina. A imunodeteccão de marcadores celulares foi avaliada em amostras prostáticas de bovinos com hiperplasia ou hiperplasia-metaplasia epiteliais, induzidas por estrógenos administrados ilegalmente e em próstatas de bovinos inoculados com dietilstilbestrol (DES). A técnica de recuperação antigênica permitiu o uso de tecidos fixados em formalina, por longos períodos. Foram utilizados os anticorpos monoclonais K8.12, anti-vimentina e anti-desmina para determinação de células basais coradas/epitélio hiperplásico-metaplásico, células do estroma e células musculares, respectivamente. As alterações tissulares observadas nos casos de campo e nos experimentais foram semelhantes, através do que se concluiu que houve administração ilegal de estrógenos. O teste imunohistoquímico com esses marca-dores específicos foi útil ao exame histológico da próstata, uma vez que a análise das imagens permite maior e melhor quantificação das alterações observadas. Os testes bioquí-micos, entretanto, são necessários para uma avaliação mais precisa.