PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Almeida A.P.M.G., Kommers G.D., Nogueira A.P.A., Júnior L.G.B., Marques B.M.F.P. & Lemos R.A.A. 2006. [Evaluation of the toxicity of Leucaena leucocephala (Leg. Mimosoi-deae) in sheep.] Avaliação do efeito tóxico de Leucaena leucocephala (Leg.Mimosoideae) em ovinos. Pesquisa Veterinária Brasileira 26(3):190-194. Departamento de Medicina Veterinária, Faculdade de Medicina Vetetinária e Zootecnia, Universidade Federal de Mato Grosso do Sul, Av. Senador Filinto Müller 2443, Cx.Postal 549, Campo Grande, MS 79070-900, Brazil. E-mail: eqrural@nin.ufms.br An experiment was performed to evaluate the toxicity of Leucaena leucocephala for sheep. Two ewes and six lambs were divided into seven treatments and fed the plant. The adult ewes did not show clinical signs of poisoning. The 4 to 5-month-old lambs presented partial wool loss 6 days after beginning of the administration of the plant and total wool loss after 12 or 20 days. The age of the sheep in this experiment suggests resistance of adult ewes and susceptibility of young lambs to L. leucocaephala poisoning.

• Poisonous plants Leucaena leucocephala Leguminosae Mimosoideae plant poisoning sheep diseases pathology

Abstract in Portuguese:

ABSTRACT.- Almeida A.P.M.G., Kommers G.D., Nogueira A.P.A., Júnior L.G.B., Marques B.M.F.P. & Lemos R.A.A. 2006. [Evaluation of the toxicity of Leucaena leucocephala (Leg. Mimosoi-deae) in sheep.] Avaliação do efeito tóxico de Leucaena leucocephala (Leg.Mimosoideae) em ovinos. Pesquisa Veterinária Brasileira 26(3):190-194. Departamento de Medicina Veterinária, Faculdade de Medicina Vetetinária e Zootecnia, Universidade Federal de Mato Grosso do Sul, Av. Senador Filinto Müller 2443, Cx.Postal 549, Campo Grande, MS 79070-900, Brazil. E-mail: eqrural@nin.ufms.br An experiment was performed to evaluate the toxicity of Leucaena leucocephala for sheep. Two ewes and six lambs were divided into seven treatments and fed the plant. The adult ewes did not show clinical signs of poisoning. The 4 to 5-month-old lambs presented partial wool loss 6 days after beginning of the administration of the plant and total wool loss after 12 or 20 days. The age of the sheep in this experiment suggests resistance of adult ewes and susceptibility of young lambs to L. leucocaephala poisoning.

Abstract in English:

Simionatto S., Vaz E.K., Michelon A., Seixas F.K., Dellagostin O.A. 2005. [Development and evaluation of new strategies for immunization against swine colibacillosis.] Desenvolvimento e avaliação de novas estratégias de imunização contra colibacilose suína. Pes-quisa Veterinária Brasileira 25(2):84-90. Laboratório de Biologia Molecular, Centro de Bio-tecnologia, UFPel, Campus Capão do Leão, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: ssimionatto@bol.com.br Swine colibacillosis caused by enterotoxigenic Escherichia coli remains one of the main sanitary problems in pig farms. The recombinant DNA technology offers the possibility of developing new immunization strategies. This paper describes the development of a subunit vaccine through the expression and purification of the E. coli K88 FaeC fimbrial protein. The gene that codes for this antigen was amplified by PCR and cloned into an E. coli expression vector fused to a 6X histidine tag. The recombinant protein was purified by affinity chromatography and used for mice immunization. In parallel, the same gene was cloned into an eucariotic expression vector with the addition of the Kozak sequence for improving translation of this gene in muscle cells. The resulting plasmid named pUP310 was purified in large scale and used to immunize mice. The immune response afforded by both forms of immunization was monitored by ELISA. There was an immune response in mice inoculated with pUP310 and purified FaeC. It was possible to detect anti-FaeC antibodies 42 days after the first inoculation. The antibody titer increased with time, being still detectable 7 months after the first inoculation. It is concluded that recombinant FaeC and pUP310 are potential tools for immunization of swine against E. coli K88.

• Swine colibacillosis FaeC Escherichia coli DNA vaccine recombinant vaccine.

Abstract in Portuguese:

Simionatto S., Vaz E.K., Michelon A., Seixas F.K., Dellagostin O.A. 2005. [Development and evaluation of new strategies for immunization against swine colibacillosis.] Desenvolvimento e avaliação de novas estratégias de imunização contra colibacilose suína. Pes-quisa Veterinária Brasileira 25(2):84-90. Laboratório de Biologia Molecular, Centro de Bio-tecnologia, UFPel, Campus Capão do Leão, Cx. Postal 354, Pelotas, RS 96010-900, Brazil. E-mail: ssimionatto@bol.com.br Swine colibacillosis caused by enterotoxigenic Escherichia coli remains one of the main sanitary problems in pig farms. The recombinant DNA technology offers the possibility of developing new immunization strategies. This paper describes the development of a subunit vaccine through the expression and purification of the E. coli K88 FaeC fimbrial protein. The gene that codes for this antigen was amplified by PCR and cloned into an E. coli expression vector fused to a 6X histidine tag. The recombinant protein was purified by affinity chromatography and used for mice immunization. In parallel, the same gene was cloned into an eucariotic expression vector with the addition of the Kozak sequence for improving translation of this gene in muscle cells. The resulting plasmid named pUP310 was purified in large scale and used to immunize mice. The immune response afforded by both forms of immunization was monitored by ELISA. There was an immune response in mice inoculated with pUP310 and purified FaeC. It was possible to detect anti-FaeC antibodies 42 days after the first inoculation. The antibody titer increased with time, being still detectable 7 months after the first inoculation. It is concluded that recombinant FaeC and pUP310 are potential tools for immunization of swine against E. coli K88.

Abstract in English:

Amaral L.A, Isa H., Dias L.T., Rossi Jr. O.D. & Nader Filho A. 2004. [Efficiency of the disinfection of teatcups and teats during mechanic milking of dairy cows.] Avaliação da eficiência da desinfecção de teteiras e dos tetos no processo de ordenha mecânica de vacas. Pesquisa Veterinária Brasileira 24(4):173-177. FCAV, Universidade Estadual Paulista, Via de Acesso Prof. Paulo Donato Castellane s/n, Jaboticabal, SP 14884-900, Brazil. E-mail: lamaral@fcav.unesp.br The purpose of this investigation was to evaluate the efficiency of the teatcup disinfection process by immersion into a disinfectant solution, using sodium hypochlorite and organic chlorine, to verify the residual chlorine dynamics in both solutions during milking time, and to evaluate three premilking disinfection methods: immersion, spray and sponge. Total coliforms, fecal coliforms, Staphylococcus sp and mesophilic microorganism counting was done. The results showed that the disinfecting clusters between individual cows, using immersion in sodium hypochlorite or organic chlorine solutions with a concentration of 150ppm, was not efficient for the rubber teatcup microorganism reduction. The organic chlorine solution showed more stability. Teatcup is a potential microorganism vehicle for milk and the mammary gland. Disinfection of the teats reduced the number of the microorganisms studied in all treatments, showing to be a simple method that can be used to reduce the microrganism transmission risks during milking and to increase the microbial quality of the milk.

• Disinfection milking chlorine teatcups pre-dipping.

Abstract in Portuguese:

Amaral L.A, Isa H., Dias L.T., Rossi Jr. O.D. & Nader Filho A. 2004. [Efficiency of the disinfection of teatcups and teats during mechanic milking of dairy cows.] Avaliação da eficiência da desinfecção de teteiras e dos tetos no processo de ordenha mecânica de vacas. Pesquisa Veterinária Brasileira 24(4):173-177. FCAV, Universidade Estadual Paulista, Via de Acesso Prof. Paulo Donato Castellane s/n, Jaboticabal, SP 14884-900, Brazil. E-mail: lamaral@fcav.unesp.br The purpose of this investigation was to evaluate the efficiency of the teatcup disinfection process by immersion into a disinfectant solution, using sodium hypochlorite and organic chlorine, to verify the residual chlorine dynamics in both solutions during milking time, and to evaluate three premilking disinfection methods: immersion, spray and sponge. Total coliforms, fecal coliforms, Staphylococcus sp and mesophilic microorganism counting was done. The results showed that the disinfecting clusters between individual cows, using immersion in sodium hypochlorite or organic chlorine solutions with a concentration of 150ppm, was not efficient for the rubber teatcup microorganism reduction. The organic chlorine solution showed more stability. Teatcup is a potential microorganism vehicle for milk and the mammary gland. Disinfection of the teats reduced the number of the microorganisms studied in all treatments, showing to be a simple method that can be used to reduce the microrganism transmission risks during milking and to increase the microbial quality of the milk.

Abstract in English:

Paulovich F.B., Borowski S.M., Driemeier D., Razia L.E., Coutinho T. A., Prates A.B.H., Pescador C., Correa A. & Barcellos D.E.S.N. 2004. [Assessment of the pathogenicity of Brachyspira pilosicoli strains by conventional histopathological techniques and by immunohistochemistry.] Avaliação da patogenicidade de amostras de Brachyspira pilosicoli através de técnicas histopatológicas convencionais e por imuno-histoquímica. Pesquisa Veterinária Brasileira 24(3):144-148. Depto Patologia Clínica Veterinária, Faculdade de Veterinária, UFRGS, Cx. Postal 15094, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@vortex.ufrgs.br The study assessed differences of pathogenicity among 19 strains of Brachyspira pilosicoli isolated from cases of diarrhea in swine in the State of Rio Grande do Sul, Brazil, using an one-day-old chick model. The chicken inoculated with live bacterial cultures were submitted to euthanasia 21 days later, and the cecum was examined histologically using hematoxylin-eosin, silver staining and immunohistochemistry with a polyclonal antibody anti-Brachyspira pilosicoli. With silver staining and immunohistochemistry, respectively, 21.59% and 70.96% of the chicken showed colonization of the cecal epithelium with B. pilosicoli. Differences in the type of colonization characterized by continuous adhesion, focal adhesion or free bacteria in the intestinal lumen were observed. Immunohistochemistry was more efficient to assess intestinal colonization, because 49.37% more colonized chicken could be detected with silver staining. With three strains, elongated figures inside the cytoplasm of epithelial cecal cells in the inoculated chicken were observed.

• Brachyspira spp Brachyspira pilosicoli spirochetal colitis immunohistochemistry pathogenesis.

Abstract in Portuguese:

Paulovich F.B., Borowski S.M., Driemeier D., Razia L.E., Coutinho T. A., Prates A.B.H., Pescador C., Correa A. & Barcellos D.E.S.N. 2004. [Assessment of the pathogenicity of Brachyspira pilosicoli strains by conventional histopathological techniques and by immunohistochemistry.] Avaliação da patogenicidade de amostras de Brachyspira pilosicoli através de técnicas histopatológicas convencionais e por imuno-histoquímica. Pesquisa Veterinária Brasileira 24(3):144-148. Depto Patologia Clínica Veterinária, Faculdade de Veterinária, UFRGS, Cx. Postal 15094, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@vortex.ufrgs.br The study assessed differences of pathogenicity among 19 strains of Brachyspira pilosicoli isolated from cases of diarrhea in swine in the State of Rio Grande do Sul, Brazil, using an one-day-old chick model. The chicken inoculated with live bacterial cultures were submitted to euthanasia 21 days later, and the cecum was examined histologically using hematoxylin-eosin, silver staining and immunohistochemistry with a polyclonal antibody anti-Brachyspira pilosicoli. With silver staining and immunohistochemistry, respectively, 21.59% and 70.96% of the chicken showed colonization of the cecal epithelium with B. pilosicoli. Differences in the type of colonization characterized by continuous adhesion, focal adhesion or free bacteria in the intestinal lumen were observed. Immunohistochemistry was more efficient to assess intestinal colonization, because 49.37% more colonized chicken could be detected with silver staining. With three strains, elongated figures inside the cytoplasm of epithelial cecal cells in the inoculated chicken were observed.

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.- Molnár L., Molnár E., Lima E.S.C. & Dias H.L.T. 2002. [Evaluation of six serological tests for the diagnosis of brucellosis in water buffaloes.] Avaliação de seis testes sorológicos no diagnóstico da brucelose bubalina. Pesquisa Veterinária Brasileira 22(2):41-44. Centro Agropecuário, Lidea, Universidade Federal do Pará, Belém, PA 66075-900, Brazil. Four hundred and forty buffalo sera, selected from about 1,200 blood samples of another study, were examined. The samples were tested by six serological methods: two of agglutination, two of indirect ELISA and two of competitive ELISA. To determine the relative sensitivity and specificity of different tests, animals with a positive result to competitive ELISA of the FAO/IAEA were considered as infected. The relative sensitivity of competitive ELISA, indirect ELISA with conjugate anti-bovine light chain monoclonal antibody labelled with HRPO, indirect ELISA with anti-bovine lgG conjugate, rose Bengal test and rapid slide agglutination test was 100%, 98.57%, 97.14%, 91.42% and 79.28%, and the relative specificity 99.33%, 97.33%, 95.66%, 94.00% and 86.33%, respectively. The value of the different serological tests for the diagnosis of brucellosis is discussed.

• Brucellosis water buffalo competitive ELISA indirect ELISA agglutination Bucelose bubalino ELISA competitivo ELISA indireto aglutinação.

Abstract in Portuguese:

RESUMO.- Molnár L., Molnár E., Lima E.S.C. & Dias H.L.T. 2002. [Evaluation of six serological tests for the diagnosis of brucellosis in water buffaloes.] Avaliação de seis testes sorológicos no diagnóstico da brucelose bubalina. Pesquisa Veterinária Brasileira 22(2):41-44. Centro Agropecuário, Lidea, Universidade Federal do Pará, Belém, PA 66075-900, Brazil. Foram examinados 440 soros bubalinos, selecionados em um outro exame de cerca de 1200 amostras sangüíneas. Utilizaram-se seis diferentes testes sorológicos para o exame dessas amostras: dois de aglutinação, dois de ELISA indireto e dois de ELISA competitivo. Os animais positivos no ELISA competitivo da FAO/IAEA foram considerados como infectados, e a comparação com os resultados dos outros testes aconteceu neste sentido. A sensibilidade relativa foi de 100%, 98,57%, 97,14%, 91,42% e 79,28%, e a especificidade relativa de 99,33%, 97,33%, 95,66%, 94,00% e 86,33% nas provas de ELISA competitivo, ELISA indireto com conjugado antibovino de cadeia leve (anticorpo monoclonal com HRPO), ELISA indireto com conjugado contra lgG bovino total, teste do antígeno acidificado tamponado e aglutinação rápida, respectivamente. Discute-se o valor dos diferentes testes sorológicos no diagnóstico da brucelose.

Abstract in English:

ABSTRACT.- Schenk M.A.M., Mendonça C.L., Madruga C.R., Kohayagawa A. & Araújo F.R. 2001. [Clinical and laboratorial evaluation of Nellore cattle experimentally infected with Trypanosoma vivax] Avaliação clínico and laboratorial de bovinos nelore infectados experimentalmente com Trypanosoma vivax. Pesquisa Veterinária Brasileira 21(4):157-161. Embrapa Gado de Corte, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. In arder to evaluate the clinical-laboratorial alterations, six Neil ore calves were inoculated with 107 Trypanosoma vivax isolated from Poconé region, Mato Grosso, Brazil. The animals were evaluated daily for rectal temperature, packed cell volume (PCV), parasitemia, antibody production, calor of mucous membranes, behavior and appetite. Blood and serum samples for biochemical evaluation for aspartate aminotransferase (AST), alkaline phbsphatase (AF), gamma glutamyltransferase (GGT), cholesterol, urea, creatinine, creatine kinase (CK), calcium, phosphorus and proteinogram were collected on days 4, 8, 12, 16, 23 and 30 post inoculation (DPI). During the following 6 months rectal temperature, PCV and parasitemia were evaluated weekly. T. vivax was evidenced from 1 DPI in all calves and persisted until day 30 in five of six animals. A remarkable decrease (p<0.05) of PCV mean value (25%) was observed on 10 DPI. The animals presented no alterations in their clinical or serum biochemical state during the trial. Seroconversion took place 6 and 8 DPI, and all the animals remained seropositive during the 30 days of experiment. In all the experimental animals the occurrence of T. vivax infection was verified, characterized by the increase of corporal temperature, presente of the blood protozoa and reduction of the globular volume, without altéations in the other variables analyzed. Nellore calves, when experimentally inoculated with T. vivax, are able to establish a balance between host-parasite relationship.

• Trypanosoma vivax calves hematology serum biochemistry proteinogram bezerros hematologia bioquímica sérica proteinograma.

Abstract in Portuguese:

RESUMO.- Schenk M.A.M., Mendonça C.L., Madruga C.R., Kohayagawa A. & Araújo F.R. 2001. [Clinical and laboratorial evaluation of Nellore cattle experimentally infected with Trypanosoma vivax] Avaliação clínico and laboratorial de bovinos nelore infectados experimentalmente com Trypanosoma vivax. Pesquisa Veterinária Brasileira 21(4):157-161. Embrapa Gado de Corte, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. Avaliaram-se as alterações clínico-laboratoriais de seis bezerros Nelore, de ambos os sexos, inoculados experimentalmente com 107 organismos viáveis de Trypanosoma vivax, isolados de bovinos da região de Poconé, Estado de Mato Grosso. Os animais foram observados diariamente, durante 30 dias, quanto aos parâmetros de temperatura retal, volume globular (VG), parasitemia, produção de anticorpos, coloração de mucosas, comportamento e apetite. Deteminaram-se os níveis séricos de aspartato aminotransferase (AST), fosfatase alcalina (FA), gama glutamiltransferase (GGT), creatina kinase (CK), colesterol, uréia, creatinina, cálcio, fósforo e o perfil eletroforético das proteínas séricas aos 4, 8, 12, 16, 23 e 30 dias pós-inoculação (DPI). Durante os 6 meses seguintes, os animais foram observados semanalmente, avaliando-se a temperatura retal, o VG e a parasitemia. T. vivax foi evidenciado a partir do terceiro e quarto DPI em todos os bezerros e persistiu até o 30º DPI em cinco dos seis animais em estudo. Ocorreu um decréscimo significativo (p<0,05) do valor médio do VG (25%) aos dez DPI. Os animais não apresentaram qualquer alteração no quadro clínico, bem como na avaliação da bioquímica sérica durante o período experimental. A soroconversão ocorreu aos 6 e 8 DPI, permanecendo todos os animais soropositivos nos 30 dias experimentais. Bovinos nelores jovens, infectados experimentalmente com T. vivax, foram capazes de estabelecer um equilíbrio na relação hospedeiro-parasita.

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., 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.

Abstract in English:

ABSTRACT.- Moraes S.S. 1998. [Evaluation of zinc, manganese and iron levels in liver samples of cattle and sheep from various regions of Brazil.] Avaliação das concentrações de zinco, manganês e ferro no fígado de bovinos e ovinos de várias regiões do Brasil. Pesquisa Veterinária Brasileira 18(3/4):107-110. Centro Nacional de Pesquisa de Gado de Corte, Embrapa, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. To complement studies on diseases of cattle and sheep possibly caused by mineral deficiencies, liver samples collected· in several regions of Brazil and which had been analysed for copper and cobalt were now analysed for zinc, manganese and iron. In most regions the levels of zinc were only sporadically low. Only in the States of Ceará and Piauí a greater number of samples had low levels of this mineral, suggesting the occurrence of zinc deficiency, but subclinically, as case histories and clinical pathological studies did not indicate the occurrence of this deficiency. Also the values for manganese were sporadically Iower than those considered normal. Only in the States of Rio de Janeiro (southern part), Espírito Santo (northern part), Ceará (Serra da Ibiapaba) and Amapá, there was a greater incidence of low values of manganese; the sarne reasons mentioned for zinc indicate that deficiency of manganese in these regions occurs subclinically.

• Mineral deficiencies zinc and manganese deficiency cattle sheep Brazil Deficiências minerais deficiências de zinco e manganês bovinos ovinos.

Abstract in Portuguese:

RESUMO.- Moraes S.S. 1998. [Evaluation of zinc, manganese and iron levels in liver samples of cattle and sheep from various regions of Brazil.] Avaliação das concentrações de zinco, manganês e ferro no figado de bovinos e ovinos de várias regiões do Brasil. Pesquisa Veterinária Brasileira 18(3/4):107-110. Centro Nacional de Pesquisa de Gado de Corte, Embrapa, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. Complementando estudos realizados sobre doenças de bovinos e ovinos possivelmente causadas por deficiências minerais, as amostras de fígado coletadas em diversas regiões do Brasil, que tinham sido analisadas somente para cobre e cobalto, foram agora analisadas para zinco, manganês e ferro. Na maioria das regiões, os valores de zinco só esporadicamente estiveram abaixo do nível considerado normal. Somente dos Estados do Ceará e do Piauí, número maior de amostras apresentou teores baixos para este mineral, sugerindo a ocorrência de deficiência de zinco, mas sob a forma sub-clínica, pois os históricos e estudos clínico-patológicos não indicavam a ocorrência desta deficiência. Os valores de manganês também só esporadicamente estiveram abaixo do nível considerado normal. Somente nas amostras dos Estados do Rio de Janeiro (Sul), do Espírito Santo (Norte), do Ceará (Serra da lbiapaba) e do Amapá, havia uma incidência maior de valores baixos de manganês; os mesmos motivos apontados acima para o zinco, indicam que a deficiência de manganês nestas regiões deve ocorrer sob forma sub-clínica.