PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Carvalho D., Oliveira T.M.F.S., Baldani C.D. & Machado R.Z. 2009. An enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies against Leishmania chagasi in dogs. Pesquisa Veterinária Brasileira 29(2):120-124. Departamento de Patologia Veterinária, Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, Via de Acesso Prof. Paulo Donato Castellane s/n, Jaboticabal, SP 14870-000, Brazil. E-mail: zacarias@fcav.unesp.br Visceral leishmaniasis is an emergent zoonosis with an increasing number of new cases in Brazil where the domestic dog is an important parasite reservoir in the infectious cycle of Leishmania chagasi. An enzyme-linked immunosorbent assay (ELISA), based upon the use of a total soluble antigenic preparation of L. chagasi, was adapted for the detection of IgM antibodies in the serum of infected dogs. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkboard titrations. The specificity and sensitivity of the ELISA were 100 %. A total of 110 serum samples were taken from dogs in Belo Horizonte, Minas Gerais, Brazil, and examined for anti-L. chagasi IgM antibody by ELISA and indirect fluorescent antibody test (IFAT). About 25% (n=27) of all the dogs tested were found serologically positive for L. chagasi by IFAT, while 89.09% (n=98) were seropositive by ELISA. The results obtained by ELISA and IFAT were significantly different (P<0.01). The combined use of ELISA and IFAT is recommended in order to enable veterinary services to more efficiently detect canine visceral leishmaniasis.

• Leishmania chagasi diagnosis dogs IgM enzyme-linked immunosorbent assay

Abstract in Portuguese:

ABSTRACT.- Carvalho D., Oliveira T.M.F.S., Baldani C.D. & Machado R.Z. 2009. An enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies against Leishmania chagasi in dogs. Pesquisa Veterinária Brasileira 29(2):120-124. Departamento de Patologia Veterinária, Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, Via de Acesso Prof. Paulo Donato Castellane s/n, Jaboticabal, SP 14870-000, Brazil. E-mail: zacarias@fcav.unesp.br Visceral leishmaniasis is an emergent zoonosis with an increasing number of new cases in Brazil where the domestic dog is an important parasite reservoir in the infectious cycle of Leishmania chagasi. An enzyme-linked immunosorbent assay (ELISA), based upon the use of a total soluble antigenic preparation of L. chagasi, was adapted for the detection of IgM antibodies in the serum of infected dogs. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkboard titrations. The specificity and sensitivity of the ELISA were 100 %. A total of 110 serum samples were taken from dogs in Belo Horizonte, Minas Gerais, Brazil, and examined for anti-L. chagasi IgM antibody by ELISA and indirect fluorescent antibody test (IFAT). About 25% (n=27) of all the dogs tested were found serologically positive for L. chagasi by IFAT, while 89.09% (n=98) were seropositive by ELISA. The results obtained by ELISA and IFAT were significantly different (P<0.01). The combined use of ELISA and IFAT is recommended in order to enable veterinary services to more efficiently detect canine visceral leishmaniasis.

Abstract in English:

ABSTRACT.- Diéguez F.J., Yus E., Sanjuán M.L., Vilar M.J. & Arnaiz I. 2008. Monitoring bovine viral diarrhea virus (BVDV) infection status in dairy herds. Pesquisa Veterinária Brasileira 28(12):588-592. Unidad de Epidemiología y Sanidad Animal, Instituto de Investigación y Análisis Alimentarios, Facultad de Veterinaria s/n, Lugo 27002, Spain. E-mail: franciscojavier.dieguez@usc.es This study was designed to assess the relationship between antibodies against bovine viral diarrhea virus (BVDV) determined in the bulk tank milk (BTM) and the within-herd seroprevalence. We also assessed the efficiency of measuring antibody levels in BTM samples to monitor BVDV infection status in a herd. In the 81 farms included in the study, BTM samples were obtained and blood samples withdrawn from all cattle older than one year. The infection status was then determined in serum and milk using a commercial blocking ELISA based on the detection of anti-p80 antibodies. Apart from these baseline serum and milk samples, another BTM sample was collected from each herd 9 months later, and a third BTM sample obtained 9 months after this. In these second and third milk samples, anti-BVDV antibodies were determined using the same ELISA kit. Statistical tests revealed good agreement between herd seroprevalences (% seropositive animals in the herd) and the antibody levels detected in the BTM samples. During the 18 months of follow-up, the farms with persistently infected cattle at the study outset (14.8% of the herds) showed a significant decrease in BTM antibody titers after virus clearance. Conversely, a significant increase in BTM antibody levels was observed in the herds infected with BVDV during the follow-up period. Our findings indicate that monitoring antibody levels in the BTM is a useful method of identifying changes in the BVDV infection status of a herd.

• Bovine viral diarrhea vírus BVDV antibodies diagnostic milk

Abstract in Portuguese:

ABSTRACT.- Diéguez F.J., Yus E., Sanjuán M.L., Vilar M.J. & Arnaiz I. 2008. Monitoring bovine viral diarrhea virus (BVDV) infection status in dairy herds. Pesquisa Veterinária Brasileira 28(12):588-592. Unidad de Epidemiología y Sanidad Animal, Instituto de Investigación y Análisis Alimentarios, Facultad de Veterinaria s/n, Lugo 27002, Spain. E-mail: franciscojavier.dieguez@usc.es This study was designed to assess the relationship between antibodies against bovine viral diarrhea virus (BVDV) determined in the bulk tank milk (BTM) and the within-herd seroprevalence. We also assessed the efficiency of measuring antibody levels in BTM samples to monitor BVDV infection status in a herd. In the 81 farms included in the study, BTM samples were obtained and blood samples withdrawn from all cattle older than one year. The infection status was then determined in serum and milk using a commercial blocking ELISA based on the detection of anti-p80 antibodies. Apart from these baseline serum and milk samples, another BTM sample was collected from each herd 9 months later, and a third BTM sample obtained 9 months after this. In these second and third milk samples, anti-BVDV antibodies were determined using the same ELISA kit. Statistical tests revealed good agreement between herd seroprevalences (% seropositive animals in the herd) and the antibody levels detected in the BTM samples. During the 18 months of follow-up, the farms with persistently infected cattle at the study outset (14.8% of the herds) showed a significant decrease in BTM antibody titers after virus clearance. Conversely, a significant increase in BTM antibody levels was observed in the herds infected with BVDV during the follow-up period. Our findings indicate that monitoring antibody levels in the BTM is a useful method of identifying changes in the BVDV infection status of a herd.

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:

Canal C.W., Ferreira D.J., Macagnan M., Fallavena L.C.B., Moraes H.L.S. & Wald V.B. 2004. Prevalence of antibodies against chicken anaemia virus (CAV) in broiler breeders in Southern Brazil. Pesquisa Veterinária Brasileira 24(2):89-92. Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Faculdade de Veterinária da Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91540-000, Brazil. E-mail: claudio.canal@ufrgs.br Chicks infected during the first two weeks of life with chicken anaemia virus (CAV) manifest clinical disease that can be avoided if the breeder hens transfer enough antibodies to their progeny. The objective of the present work was to establish the prevalence and titer of anti-CAV antibodies in some Brazilian broiler hen breeder flocks and verify in which phase of life the birds were infected. A total of 1,709 serum samples from 12 broiler hen flocks vaccinated against CAV and 64 unvaccinated flocks were analyzed for CAV antibodies with an enzyme-linked immunosorbent assay (ELISA). All non-vaccinated breeder flocks were found to be infected with CAV, with 89% of the hens tested presenting antibodies, 52% of these with titers considered high enough to protect their progeny against CAV infection. Likewise, all vaccinated hens had antibody titer to CAV capable of conferring protection to their progeny. Thus, vaccination of hens seems capable of conferring protection to chicks against clinically apparent CAV-associated disease.

• Antibodies avian pathology chicken anemia virus CAV epidemiology prevalence.

Abstract in Portuguese:

Canal C.W., Ferreira D.J., Macagnan M., Fallavena L.C.B., Moraes H.L.S. & Wald V.B. 2004. Prevalence of antibodies against chicken anaemia virus (CAV) in broiler breeders in Southern Brazil. Pesquisa Veterinária Brasileira 24(2):89-92. Centro de Diagnóstico e Pesquisa em Patologia Aviária (CDPA), Faculdade de Veterinária da Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91540-000, Brazil. E-mail: claudio.canal@ufrgs.br Chicks infected during the first two weeks of life with chicken anaemia virus (CAV) manifest clinical disease that can be avoided if the breeder hens transfer enough antibodies to their progeny. The objective of the present work was to establish the prevalence and titer of anti-CAV antibodies in some Brazilian broiler hen breeder flocks and verify in which phase of life the birds were infected. A total of 1,709 serum samples from 12 broiler hen flocks vaccinated against CAV and 64 unvaccinated flocks were analyzed for CAV antibodies with an enzyme-linked immunosorbent assay (ELISA). All non-vaccinated breeder flocks were found to be infected with CAV, with 89% of the hens tested presenting antibodies, 52% of these with titers considered high enough to protect their progeny against CAV infection. Likewise, all vaccinated hens had antibody titer to CAV capable of conferring protection to their progeny. Thus, vaccination of hens seems capable of conferring protection to chicks against clinically apparent CAV-associated disease.

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.- Souza V.F, Melo S.V., Esteves P.A., Schmidt C.S.R., Gonçalves D.A., Schaefer R., Silva T.C., Almeida R.S., Vicentini F., Franco A.C., Oliveira E.A., Spilki F.R., Weiblen R., Flores E.F., Lemos R.A., Alfieri A.A., Pituco E.M. & Roehe P.M. 2002. [Monoclonal antibody characterization of bovine herpesviruses types 1 (BHV-1) and 5 (BHV-5).] Caracterização de herpesvírus bovinos tipos 1 (BHV-1) e 5 (BHV-5) com anticorpos monoclonais. Pesquisa Veterinária Brasileira 22(1):13-18,-Depto Microbiologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, and Centro de Pesquisas Universitárias Desidério Finamor, Estrada do Conde 6000, Eldorado do Sul, RS 90001-970, Brazíl. E-mail: proehe@vottex.ufrgs.br The antigenic profile of 45 herpesviruses (44 viruses from cattle, including six reference BHV-1 strains and 15 putative BHV-1; three reference BHV-5 strains and 20 putative BHV-5) and one butfalo isolate (BuHV) were examined with a panei of monoclonal antibodies (Mabs) prepared against bovine herpesvirus antigens. Tests were performed by immunoperoxidase (IPX) on infected cell cultures, with the Mabs as primary antibodies. lmmunostaining allowed the differentiation between types 1 and 5 viruses. Ali isolates from cases of encephalitis displayed BHV-5 profiles. Four BHV-5 isolates obtained from geographically distinct áreas displayed different and highly variable IPX patterns of reactivity. Two viruses with BHV-5 antigenic profile were isolated from semen of asymptomatic bulis. The results showed that the antigenic characterization with the Mab panei employed here is useful for typing BHV-1 and BHV-5 isolates.

• Bovine herpesvirus BHV-1 BHV-5 monoclonal antibodies immunoperoxidase Herpesvírus bovino 1 herpesvírus bovino 5 anticorpos monoclonais imunoperoxidase.

Abstract in Portuguese:

RESUMO.- Souza V.F, Melo S.V., Esteves P.A., Schmidt C.S.R., Gonçalves D.A., Schaefer R., Silva T.C., Almeida R.S., Vicentini F., Franco A.C., Oliveira E.A., Spilki F.R., Weiblen R., Flores E.F., Lemos R.A., Alfieri A.A., Pituco E.M. & Roehe P.M. 2002. [Monoclonal antibody characterization of bovine herpesviruses types 1 (BHV-1) and 5 (BHV-5).] Caracterização de herpesvírus bovinos tipos 1 (BHV-1) e 5 (BHV-5) com anticorpos monoclonais. Pesquisa Veterinária Brasileira 22(1):13-18,-Depto Microbiologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, and Centro de Pesquisas Universitárias Desidério Finamor, Estrada do Conde 6000, Eldorado do Sul, RS 90001-970, Brazíl. E-mail: proehe@vottex.ufrgs.br O perfil antigénico de 45 herpesvírus (44 de bovinos, sendo seis amostras de referência de BHV-1 e 15 prováveis BHV-1; três amostras de referência de BHV-5 e 20 prováveis BHV-5) e uma amostra de herpesvírus bubalino (BuHV) foi examinado com um painel de anticorpos monoclonais (Acms) produzidos contra antígenos de herpesvírus bovinos. Para os exames, foi utilizada a prova de imunoperoxidase (IPX) sobre cultivas de células infectadas, tendo os Acms como anticorpos primários. A determinação dos padrões de reatividade das amostras de vírus frente aos Acms permitiu a diferenciação entre os tipos 1 e 5. Todas as amostras isoladas de casos de encefalite apresentaram perfil de BHV-5. Quatro amostras de BHV-5 isoladas de áreas geograficamente distintas apresentaram perfis de reatividade diferenciados em relação às demais amostras do tipo 5. Duas amostras de vírus com perfil antigénico de BHV-5 foram isoladas de sêmen de animais infectados. Estes resultados comprovam a utilidade da caracterização antigénica com este painel de Acms na tipagem de amostras de BHV-1 e BHV-5.

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&#39;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&#39; 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&#39;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.- Madruga C.R., Marques A.P.C., Leal C.R.B., Carvalho C.M.E., Araújo F.R. & Kessler R.H. 2000. Evaluation of an enzyme-linked immunosorbent assay to detect antibodies against Anaplasma marginale. [Avaliação de uma prova de imunoadsorção enzimática para detecção de anticorpos contra Anaplasma marginale.] Pesquisa Veterinária Brasileira 20(3):109-112. Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. A rapid indirect enzyme-linked immunosorbent assay (ELISA) was developed for measuring antibodies against Anaplasma marginale using a partially soluble antigen prepared from semipurified initial bodies from erythrocytes with 80.0% of rickettsiaemia. This technique utilized alkaline phosphatase and p-nitrophenyl phosphate as reaction indicators. The high sensitivity (100.0%) was confirmed with sera from 100 calves experimentally-infected with A. marginale. All of these animals showed seroconversion before or at the same time of the first rickettsiaemia or even when it was not detected. Also the elevated specificity (94.0%) was confirmed by the low percentage of cross-reactions with sera from animals experimentally-infected with Babesia bigemina and Babesia bovis (1.4 and 6.6%, respectively). Performances of ELISA and indirect fluorescent antibody test (IFAT) with 324 sera from enzootically stable area did not show statistical difference (P>0.05), since the former showed 96.9% and the latter 97.2% of positive reactions. The advantage of this ELISA is a shorter execution time than others developed until now, allowing more samples to be analyzed.

• Anaplasma marginale ELISA serological tests cattle testes sorológicos bovino.

Abstract in Portuguese:

RESUMO.- Madruga C.R., Marques A.P.C., Leal C.R.B., Carvalho C.M.E., Araújo F.R. & Kessler R.H. 2000. Evaluation of an enzyme-linked immunosorbent assay to detect antibodies against Anaplasma marginale. [Avaliação de uma prova de imunoadsorção enzimática para detecção de anticorpos contra Anaplasma marginale.] Pesquisa Veterinária Brasileira 20(3):109-112. Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. Um ensaio de imunoadsorção enzimática (ELISA) indireto, foi desenvolvido para detecção de anticorpos contra Anaplasma marginale, utilizando um antígeno parcialmente solúvel preparado a partir de corpúsculos iniciais semipurificados de eritrócitos com 80,0% de parasitemia. Esta técnica utilizou a fosfatase alcalina e o p-nitrofenil fosfato como indicadores de reação. A alta sensibilidade (100,0%) foi comprovada com soros ele 100 bezerros experimentalmente infectados com A. marginale. Todos estes animais soroconverteram antes ou ao mesmo tempo que a primeira parasitemia foi detectada, ou mesmo quando não se obteve parasitemia. A elevada especificidade (94,0%) foi confirmada pela baixa percentagem ele reações cruzadas com soros de animais infectados experimentalmente com Babesia bigemina e Babesia bovis (1,4 e 6,6%, respectivamente). Os desempenhos do ELISA e da imunofluorescência indireta (IFI) com 324 soros de uma área de estabilidade enzoótica não diferiram estatisticamente (P>0,05), uma vez que o primeiro teste detectou 96,9% de reações positivas, enquanto o segundo, 97,2%. A vantagem deste ELISA foi um menor tempo ele execução do que outros desenvolvidos até o momento, permitindo a análise de um maior·número de soros.