PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Nunes Rangel M.F., Lemos L.S., Almeida C.M.C., Sales L.G. & Vieira-da-Motta O. 2010. Development of Polyclonal IgY antibodies in chickens and IgG in rabbits immunized against proteins of Pythium insidiosum isolated from horses in the state of Rio de Janeiro. Pesquisa Veterinária Brasileira 30(1):87-93. Micologia Veterinária do Setor de Doenças Infectocontagiosas, Laboratório de Sanidade Animal do Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Av. Alberto Lamego 2000, Horto, Campos dos Goytacazes, RJ 28013-602, Brazil. E-mail: olney@uenf.br Pythiosis is caused by Pythium insidiosum and the occurrence of disease in horses was described in the North and Northwest State of Rio de Janeiro, Brazil. The disease was described in cattle, sheep, humans, and horses in different states and regions across the country. This paper describes the development of IgY and IgG polyclonal antibodies, in chicken and rabbits, respectively against proteins extracted from kunkers and hyphae of P. insidiosum from affected horses. The proteins were recognized by chicken, rabbit and horse antibodies by immunodiffusion and Western blot against majority bands of 27 and 43 KDa, and titrated by ELISA. The antibodies IgY developed by the first time against Brazilian strains of P. insidiosum may represent a valuable tool in the detection of antigens of the pathogen and contribute to further studies aimed at immunotherapy and knowledge about this disease in endemic areas in Rio de Janeiro and in Brazil.

• Pythiosis Pythium insidiosum

Abstract in Portuguese:

RESUMO.- Nunes Rangel M.F., Lemos L.S., Almeida C.M.C., Sales L.G. & Vieira-da-Motta O. 2010. Development of Polyclonal IgY antibodies in chickens and IgG in rabbits immunized against proteins of Pythium insidiosum isolated from horses in the state of Rio de Janeiro. [Desenvolvimento de Anticorpos IgY em galinhas e IgG em coelhos imunizados contra proteínas de Pythium insidiosum isolado de equinos no Estado do Rio de Janeiro, Brasil.] Pesquisa Veterinária Brasileira 30(1):87-93. Micologia Veterinária do Setor de Doenças Infectocontagiosas, Laboratório de Sanidade Animal do Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Av. Alberto Lamego 2000, Horto, Campos dos Goytacazes, RJ 28013-602, Brazil. E-mail: olney@uenf.br Pitiose é causada por Pythium insidiosum e a doença foi descrita em equinos no Norte e Noroeste do Estado do Rio de Janeiro, Brasil. A doença foi descrita em bovinos, ovelhas, humanos e cavalos em diferentes estados e regiões do país. Este trabalho descreve o desenvolvimento de anticorpos policlonais, IgY e IgG, em galinha e coelho, respectivamente, contra proteínas extraídas de kunkers e hifas de P. insidiosum de cavalos doentes. As proteínas foram reconhecidas por anticorpos de galinha, coelho e cavalos contra as bandas majoritárias de 27 e 34 KDa em imunodifusão e Western blot tituladas por ELISA. Os anticorpos IgY desenvolvidos pela primeira vez contra cepas brasileiras de P. insidiosum podem representar um valioso instrumento na detecção de antígenos de patógenos e contribuem para novos estudos baseados na imunoterapia e no entendimento sobre esta doença em áreas endêmicas no Rio de Janeiro e em todo o país.

Abstract in English:

ABSTRACT.- Arduino G.G.C., Girio R.J.S., Magajevski F.S. & Pereira G.T. 2009. [Agglutinating antibody titers induced by commercial vaccines against bovine leptospirosis.] Títulos de anticorpos aglutinantes induzidos por vacinas comerciais contra leptospirose bovina. Pesquisa Veterinária Brasileira 29(7):575-582. Departamento de Medicina Veterinária Preventiva e Reprodução Animal, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Via de Acesso Paulo Donato Castellane s/n, Jaboticabal, SP 14884-900, Brazil. E-mail: cocaarduino@yahoo.com.br In the investigation 100 heifers were used, divided into 5 groups of 20 animals each. The four experimental groups were vaccinated using distinct commercial polyvalent bacterines: A, B, C and D, and a group was the control. Samples were collected at days 0, 3, 7, 14, 21, 28, 35, 42, 42, 56, 63, 70, 77, 84, 91, 120, 150 and 180 from the first injection of the vaccine. The selection of the animals for the experimental groups was done based on a serological screening with 6 antigens of Leptospira sp. constituted by non-reagent animals. The vaccine titers were monitored using the microscopic agglutination test (MAT) for Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, Pomona and Wolffi serovars. All vaccines used were capable to product agglutinins for the Hardjo and Wolffi serovars observed at 3 days after vaccination, remaining until the 150th day; those serovars induced the highest titres of agglutinins. Vaccine D, in spite of not containing the Wolffi serovar, induced the production of agglutinins to this serovar. Agglutinins to the Canicola serovar were only observed in the animals vaccinated with the D bacterine. Vaccine D induced the highest average titers of antibodies to all tested serovars.

• Leptospirosis cattle vaccine agglutinins

Abstract in Portuguese:

ABSTRACT.- Arduino G.G.C., Girio R.J.S., Magajevski F.S. & Pereira G.T. 2009. [Agglutinating antibody titers induced by commercial vaccines against bovine leptospirosis.] Títulos de anticorpos aglutinantes induzidos por vacinas comerciais contra leptospirose bovina. Pesquisa Veterinária Brasileira 29(7):575-582. Departamento de Medicina Veterinária Preventiva e Reprodução Animal, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Via de Acesso Paulo Donato Castellane s/n, Jaboticabal, SP 14884-900, Brazil. E-mail: cocaarduino@yahoo.com.br In the investigation 100 heifers were used, divided into 5 groups of 20 animals each. The four experimental groups were vaccinated using distinct commercial polyvalent bacterines: A, B, C and D, and a group was the control. Samples were collected at days 0, 3, 7, 14, 21, 28, 35, 42, 42, 56, 63, 70, 77, 84, 91, 120, 150 and 180 from the first injection of the vaccine. The selection of the animals for the experimental groups was done based on a serological screening with 6 antigens of Leptospira sp. constituted by non-reagent animals. The vaccine titers were monitored using the microscopic agglutination test (MAT) for Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, Pomona and Wolffi serovars. All vaccines used were capable to product agglutinins for the Hardjo and Wolffi serovars observed at 3 days after vaccination, remaining until the 150th day; those serovars induced the highest titres of agglutinins. Vaccine D, in spite of not containing the Wolffi serovar, induced the production of agglutinins to this serovar. Agglutinins to the Canicola serovar were only observed in the animals vaccinated with the D bacterine. Vaccine D induced the highest average titers of antibodies to all tested serovars.

Abstract in English:

ABSTRACT.- Silva-Zacarias F.G., Spohr K.A.H., Lima B.A.C., Dias J.A., Müller E.E., Ferreira Neto J.S., Turilli C. & Freitas J.C. 2009. [Prevalence of antibodies against Chlamydophila spp. in herds with bovine abortion of Paraná state, Brazil.] Prevalência de anticorpos anti-Chlamydophila spp. em propriedades rurais com histórico de aborto bovino no estado do Paraná. Pesquisa Veterinária Brasileira 29(3):215-219. Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid 380, Cx. Postal 6001, Londrina, PR 86051-990, Brazil. E-mail: freitasj@uel.br Chlamydophila abortus is a recognized cause of bovine epizootic abortion. Abortion, premature birth and weak lamb/calf, stillbirth and repeat breeding in irregular intervals are the most frequent disease manifestations. The complement fixation test is the recommended by the World Organization of Animal Health (OIE) for Chlamydophila spp. serologic diagnosis. The aim of this study was estimate the prevalence of antibodies against Chlamydophila spp. in cattle herds with abortion, selected inside the sampling design of National Program of Control and Erradication of Brucellosis in Paraná state. Serum samples of 3,102 cows (age ³ 24 months) from 373 herds were analyzed by complement fixation test. Totally, 44 (1.42%) animal were positive with titers ³ 32. The seroprevalence of Chlamydophila spp. in the herds was 8.82% (6.15%-12.17%). Four variables were associated with seroprevalence for Chlamydophila spp. in the final model of logistic regression: confined or semi-confined breeding (OR=3.339, P=0.004), farms with less than 35 cows (OR=3.339, P=0.017), abortion in the pasture (OR=2.372, P=0.037) and pasture rent (OR=3.398, P=0.006) were risk factors for Chlamydophila spp. This bacterium infected a small number of cattle from herds with abortion in Paraná state. Chlamydophila spp impact as abortion cause is reduced in this state.

• Chlamydophila abortus bovine epizootic abortion serology bacteria

Abstract in Portuguese:

ABSTRACT.- Silva-Zacarias F.G., Spohr K.A.H., Lima B.A.C., Dias J.A., Müller E.E., Ferreira Neto J.S., Turilli C. & Freitas J.C. 2009. [Prevalence of antibodies against Chlamydophila spp. in herds with bovine abortion of Paraná state, Brazil.] Prevalência de anticorpos anti-Chlamydophila spp. em propriedades rurais com histórico de aborto bovino no estado do Paraná. Pesquisa Veterinária Brasileira 29(3):215-219. Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina, Rodovia Celso Garcia Cid 380, Cx. Postal 6001, Londrina, PR 86051-990, Brazil. E-mail: freitasj@uel.br Chlamydophila abortus is a recognized cause of bovine epizootic abortion. Abortion, premature birth and weak lamb/calf, stillbirth and repeat breeding in irregular intervals are the most frequent disease manifestations. The complement fixation test is the recommended by the World Organization of Animal Health (OIE) for Chlamydophila spp. serologic diagnosis. The aim of this study was estimate the prevalence of antibodies against Chlamydophila spp. in cattle herds with abortion, selected inside the sampling design of National Program of Control and Erradication of Brucellosis in Paraná state. Serum samples of 3,102 cows (age ³ 24 months) from 373 herds were analyzed by complement fixation test. Totally, 44 (1.42%) animal were positive with titers ³ 32. The seroprevalence of Chlamydophila spp. in the herds was 8.82% (6.15%-12.17%). Four variables were associated with seroprevalence for Chlamydophila spp. in the final model of logistic regression: confined or semi-confined breeding (OR=3.339, P=0.004), farms with less than 35 cows (OR=3.339, P=0.017), abortion in the pasture (OR=2.372, P=0.037) and pasture rent (OR=3.398, P=0.006) were risk factors for Chlamydophila spp. This bacterium infected a small number of cattle from herds with abortion in Paraná state. Chlamydophila spp impact as abortion cause is reduced in this state.

Abstract in English:

ABSTRACT.- Galo K.R, Fonseca A.H., Madureira R.C. & Barbosa Neto J.D. 2009. [Occurrence of homologous anti-Borrelia burgdorferi antibodies in equines in the metropolitan mesorregion of Belém, State of Para, Brazil.] Freqüência de anticorpos homólogos anti-Borrelia burgdorferi em eqüinos na mesorregião metropolitana de Belém, Estado do Pará. Pesquisa Veterinária Brasileira 29(3):229-232. Departamento de Epidemiologia e Saúde Pública, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ 23890-000, Brazil. E-mail: adivaldo@ufrrj.br Spirochaetes transmitted by ticks are microorganisms of worldwide distribution, which infect wild, domestic animals and human beings. A total of 300 equine sera from four municipalities: Ananideua (58), Belém (61), Castanhal (131), and Santa Izabel do Pará (50), were evaluated for Borrelia burgdorferi by an Elisa test. There were no significant differences (P<0.05) among municipalities, breed, sex or husbandry. A total of 80 (26.7%) horses were B. burgdorferi positive with titles of 1:800, 72 (90%) horses, 1:1.600, 6 (7.5%) horses, and 1:3.200, 2 (2.5%) horses. The results were similar to those in the USA, where related frequencies ranged from 7 to 75% in asymptomatic seropositive horses. The presence of anti-B.burgdorferi homologous antibodies in horses from four municipalities in the metropolitan mesorregion of Belém suggests the possibility of occurrence of human cases in the region.

• Borrelia burgdorferi Spirochaetacae horses serology

Abstract in Portuguese:

ABSTRACT.- Galo K.R, Fonseca A.H., Madureira R.C. & Barbosa Neto J.D. 2009. [Occurrence of homologous anti-Borrelia burgdorferi antibodies in equines in the metropolitan mesorregion of Belém, State of Para, Brazil.] Freqüência de anticorpos homólogos anti-Borrelia burgdorferi em eqüinos na mesorregião metropolitana de Belém, Estado do Pará. Pesquisa Veterinária Brasileira 29(3):229-232. Departamento de Epidemiologia e Saúde Pública, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ 23890-000, Brazil. E-mail: adivaldo@ufrrj.br Spirochaetes transmitted by ticks are microorganisms of worldwide distribution, which infect wild, domestic animals and human beings. A total of 300 equine sera from four municipalities: Ananideua (58), Belém (61), Castanhal (131), and Santa Izabel do Pará (50), were evaluated for Borrelia burgdorferi by an Elisa test. There were no significant differences (P<0.05) among municipalities, breed, sex or husbandry. A total of 80 (26.7%) horses were B. burgdorferi positive with titles of 1:800, 72 (90%) horses, 1:1.600, 6 (7.5%) horses, and 1:3.200, 2 (2.5%) horses. The results were similar to those in the USA, where related frequencies ranged from 7 to 75% in asymptomatic seropositive horses. The presence of anti-B.burgdorferi homologous antibodies in horses from four municipalities in the metropolitan mesorregion of Belém suggests the possibility of occurrence of human cases in the region.

Abstract in English:

ABSTRACT.- Modolo J.R., Stachissini A.V.M., Gennari S.M., Dubey J.P., Langoni H., Padovani C.R., Barroso L. & Leite B.L.S. 2008. [Frequency of antibodies anti-Neospora caninum in sera of goats of the State São Paulo and its relationship with flock management.] Freqüência de anticorpos anti-Neospora caninum em soros de caprinos do estado de São Paulo e sua relação com o manejo dos animais. Pesquisa Veterinária Brasileira 28(12):597-600. Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Cx. Postal 524, Rubião Júnior s/n, Botucatu, SP 18618-000, Brazil. E-mail: jrmodolo@fmvz.unesp.br In order to assess the frequency of dairy goats seropositive for Neospora caninum in the state of São Paulo, and its relationship with age, sex, presence of dogs and reproductive problems in goat pens, serum samples were obtained from 923 goats of both sexes older than 3 months of age. Samples were collected in 17 properties located in different municipalities. Neospora agglutination test (NATe”25) was used to determine the presence of Neospora caninum in all animals. Epidemiological and reproductive data were obtained through a questionnaire which was administered in all goat pens. Significance level was set at 5% for all statistical tests. N. caninum positivity rate was 19.77%. Only one of the properties had no seropositive animal revealing N. caninum diffusion in the state. Seropositivity frequency did not correlate with sex, age, or reproductive problems. However, the presence of dogs was associated with a higher frequency of N. caninum seropositivity. A choroplethic map with hatch patterns showing the geographical distribution of goats seropositive for Neospora caninum may considerably contribute to geographic epidemiological studies and the development of disease control strategies.

• Neospora caninum occurrence; goats dogs epidemiological variables reproductive problems geographic distribution

Abstract in Portuguese:

ABSTRACT.- Modolo J.R., Stachissini A.V.M., Gennari S.M., Dubey J.P., Langoni H., Padovani C.R., Barroso L. & Leite B.L.S. 2008. [Frequency of antibodies anti-Neospora caninum in sera of goats of the State São Paulo and its relationship with flock management.] Freqüência de anticorpos anti-Neospora caninum em soros de caprinos do estado de São Paulo e sua relação com o manejo dos animais. Pesquisa Veterinária Brasileira 28(12):597-600. Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Cx. Postal 524, Rubião Júnior s/n, Botucatu, SP 18618-000, Brazil. E-mail: jrmodolo@fmvz.unesp.br In order to assess the frequency of dairy goats seropositive for Neospora caninum in the state of São Paulo, and its relationship with age, sex, presence of dogs and reproductive problems in goat pens, serum samples were obtained from 923 goats of both sexes older than 3 months of age. Samples were collected in 17 properties located in different municipalities. Neospora agglutination test (NATe”25) was used to determine the presence of Neospora caninum in all animals. Epidemiological and reproductive data were obtained through a questionnaire which was administered in all goat pens. Significance level was set at 5% for all statistical tests. N. caninum positivity rate was 19.77%. Only one of the properties had no seropositive animal revealing N. caninum diffusion in the state. Seropositivity frequency did not correlate with sex, age, or reproductive problems. However, the presence of dogs was associated with a higher frequency of N. caninum seropositivity. A choroplethic map with hatch patterns showing the geographical distribution of goats seropositive for Neospora caninum may considerably contribute to geographic epidemiological studies and the development of disease control strategies.

Abstract in English:

ABSTRACT.- Modolo J.R., Langoni H., Padovani C.R., Barrozo L.V., Leite B.L.S., Gennari S.M. & Stachissini A.V.M. 2009.[Occurrence of anti-Toxoplasma gondii antibodies in goat sera in the state of São Paulo, and its association with epidemiological variables, reproductive problems and risks on public health.] Avaliação da ocorrência de anticorpos anti-Toxoplasma gondii, em soros de caprinos do estado de São Paulo, e associação com variáveis epidemiológicas, problemas reprodutivos e riscos à saúde pública. Pesquisa Veterinária Brasileira 28(12):606-610. Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Cx. Postal 524, Botucatu, SP 18618-000, SP, Brazil. E-mail: jrmodolo@fmvz.unesp.br The study evaluated the anti-Toxplasma gondii antibodies occurrence in dairy goats in São Paulo State, and also verified likely associations with age, sex, presence of cats, occurrence of reproductive problems, and potential risks to public health. Serum was obtained from 923 animals of both sex, over three months of age from 17 properties in different cities. For the diagnosis, the indirect immunofluorescence reaction (RIFIe”16) and a questionnaire about health to collect epidemiological information and the reproductive cycle of every goat were used. Among all properties, 15 foci of T. gondii were diagnosed with seropositivity ranging from 2.70% to 81.25%. Neither association between the frequency of seropositivity and the sex of animals nor occurrence of reproduction failure of goats was identified. There was positive influence of anti-T. gondii antibody rate due to the older age of the animals and the presence of cats in pens. The risk of transmitting T. gondii to the public health must be considered, once in some properties where the positive serology was verified natural, products such as milk and/or meat are commercialized. Results were discussed at 5% level of significance.

• Toxoplasma gondii occurrence commercialization map distribution dairy goats

Abstract in Portuguese:

ABSTRACT.- Modolo J.R., Langoni H., Padovani C.R., Barrozo L.V., Leite B.L.S., Gennari S.M. & Stachissini A.V.M. 2009.[Occurrence of anti-Toxoplasma gondii antibodies in goat sera in the state of São Paulo, and its association with epidemiological variables, reproductive problems and risks on public health.] Avaliação da ocorrência de anticorpos anti-Toxoplasma gondii, em soros de caprinos do estado de São Paulo, e associação com variáveis epidemiológicas, problemas reprodutivos e riscos à saúde pública. Pesquisa Veterinária Brasileira 28(12):606-610. Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Cx. Postal 524, Botucatu, SP 18618-000, SP, Brazil. E-mail: jrmodolo@fmvz.unesp.br The study evaluated the anti-Toxplasma gondii antibodies occurrence in dairy goats in São Paulo State, and also verified likely associations with age, sex, presence of cats, occurrence of reproductive problems, and potential risks to public health. Serum was obtained from 923 animals of both sex, over three months of age from 17 properties in different cities. For the diagnosis, the indirect immunofluorescence reaction (RIFIe”16) and a questionnaire about health to collect epidemiological information and the reproductive cycle of every goat were used. Among all properties, 15 foci of T. gondii were diagnosed with seropositivity ranging from 2.70% to 81.25%. Neither association between the frequency of seropositivity and the sex of animals nor occurrence of reproduction failure of goats was identified. There was positive influence of anti-T. gondii antibody rate due to the older age of the animals and the presence of cats in pens. The risk of transmitting T. gondii to the public health must be considered, once in some properties where the positive serology was verified natural, products such as milk and/or meat are commercialized. Results were discussed at 5% level of significance.

Abstract in English:

ABSTRACT.- Melo E.S.P., Araújo F.R., Ramos C.A.N., Soares C.O., Rosinha G.M.S., Elisei C. & Madruga C.R. 2007. [ELISA based on recombinant truncated MSP5 for detection of antibodies against Anaplasma marginale in cattle.] ELISA com MSP5 recombinante truncada para detecção de anticorpos contra Anaplasma marginale em bovinos. Pesquisa Veterinária Brasileira 27(7):301-306. Embrapa Gado de Corte, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. E-mail: flabio@cnpgc.embrapa.br The objective of this study was the production and solubilization of recombinant truncated MSP5 of Anaplasma marginale and the evaluation of its performance in an enzyme-linked immunosorbent assay (ELISA), to detect antibodies against the rickettsia in cattle. The fragment of msp5 gene, except the hydrophobic N-terminal region, was amplified by PCR, cloned in pTrcHis-TOPO plasmid and expressed in Escherichia coli. Solubilization of the recombinant protein was evaluated in different pHs and concentrations of urea. The sensibility and specificity of the assay were evaluated with 66 sera from cattle experimentally-infected and 96 sera from cattle free of A. marginale defined by polymerase chain reaction for msp5 gene. Serum samples from 1,666 cattle from Brazil - states of Rio Grande do Sul (73), Mato Grosso do Sul (91), Pernambuco (86), Bahia (314) and Minas Gerais (267), Uruguay (32) and Costa Rica (803), were tested by ELISAs with recombinant truncated MSP5 and with recombinant MSP1a, and the agreement between both ELISAs was calculated. ELISA with recombinant truncated MSP5 protein detected infected animals with sensibility of 96.97% and specificity of 100%. In cattle experimentally-infected, the ELISA detected antibodies from the 12th day post-infection (DPI) to the end of the experiment, at the 37th DPI. The agreement between the ELISAs with truncated MSP5 and MSP1a antigens was 95.67%, with a kappa index of 0.81. Disagreement results showed significative difference (p <0.001). Antibodies for A. marginale were detected in animals of the all the region analyzed. The ELISA with recombinant truncated MSP5 showed a good performance in ELISA for detention of antibodies against A. marginale, with high sensitivity and specificity, representing an important tool for the diagnosis of anaplasmose bovine in epidemiological studies.

• Anaplasma marginale ELISA MSP5 solubilization recombinant protein

Abstract in Portuguese:

ABSTRACT.- Melo E.S.P., Araújo F.R., Ramos C.A.N., Soares C.O., Rosinha G.M.S., Elisei C. & Madruga C.R. 2007. [ELISA based on recombinant truncated MSP5 for detection of antibodies against Anaplasma marginale in cattle.] ELISA com MSP5 recombinante truncada para detecção de anticorpos contra Anaplasma marginale em bovinos. Pesquisa Veterinária Brasileira 27(7):301-306. Embrapa Gado de Corte, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil. E-mail: flabio@cnpgc.embrapa.br The objective of this study was the production and solubilization of recombinant truncated MSP5 of Anaplasma marginale and the evaluation of its performance in an enzyme-linked immunosorbent assay (ELISA), to detect antibodies against the rickettsia in cattle. The fragment of msp5 gene, except the hydrophobic N-terminal region, was amplified by PCR, cloned in pTrcHis-TOPO plasmid and expressed in Escherichia coli. Solubilization of the recombinant protein was evaluated in different pHs and concentrations of urea. The sensibility and specificity of the assay were evaluated with 66 sera from cattle experimentally-infected and 96 sera from cattle free of A. marginale defined by polymerase chain reaction for msp5 gene. Serum samples from 1,666 cattle from Brazil - states of Rio Grande do Sul (73), Mato Grosso do Sul (91), Pernambuco (86), Bahia (314) and Minas Gerais (267), Uruguay (32) and Costa Rica (803), were tested by ELISAs with recombinant truncated MSP5 and with recombinant MSP1a, and the agreement between both ELISAs was calculated. ELISA with recombinant truncated MSP5 protein detected infected animals with sensibility of 96.97% and specificity of 100%. In cattle experimentally-infected, the ELISA detected antibodies from the 12th day post-infection (DPI) to the end of the experiment, at the 37th DPI. The agreement between the ELISAs with truncated MSP5 and MSP1a antigens was 95.67%, with a kappa index of 0.81. Disagreement results showed significative difference (p <0.001). Antibodies for A. marginale were detected in animals of the all the region analyzed. The ELISA with recombinant truncated MSP5 showed a good performance in ELISA for detention of antibodies against A. marginale, with high sensitivity and specificity, representing an important tool for the diagnosis of anaplasmose bovine in epidemiological studies.

Abstract in English:

Alves A.L., Madureira R.C., Silva R.A., Corrêa F.N. & Botteon R.C.C.M. 2004. [Frequency of antibodies against Borrelia burgdorferi in dogs from the metropolitan region of Rio de Janeiro.] Freqüência de anticorpos contra Borrelia burgdorferi em cães na região metropolitana do Rio de Janeiro. Pesquisa Veterinária Brasileira 24 (4):203-206. Depto Medicina e Cirurgia Veterinária, Inst. Veterinária, UFRRJ, Seropédica, RJ 23851-970, Brazil. E-mail: fabiolanc@ig.com.br Lyme disease is a worldwide zoonosis caused by Borrelia burgdorferi (Spirochaetae) and transmitted through ticks (Ixodidae). Many species of domestic and wild animals are affected. The dog is an important reservoir in the domiciliary environment. The disease affects the joints and causes alterations in other organs. Blood serum samples from 143 dogs, living in the counties of Seropédica, Itaguaí and Rio de Janeiro, were analyzed with the indirect ELISA test, to search for antibodies against Borrelia burgdorferi latu sensu. Average positive results were 48.25%, 41.9% in Seropédica, 4.9% in Itaguaí and 1.4% in Rio de Janeiro. The titres varied from 1:400 to 1:6400. The prevalence was lower (p< 0.05) in dogs up to one year of age than in older dogs, with significant difference. Between males and females was no statistical difference with positives titres. The result of this survey should draw attention to the occurrence of Borrelia sp in the area studied, considering the importance of Lyme disease as an emerging zoonosis.

• Borrelia burgdorferi Spirochaetae dogs enzyme immunoassay serology.

Abstract in Portuguese:

Alves A.L., Madureira R.C., Silva R.A., Corrêa F.N. & Botteon R.C.C.M. 2004. [Frequency of antibodies against Borrelia burgdorferi in dogs from the metropolitan region of Rio de Janeiro.] Freqüência de anticorpos contra Borrelia burgdorferi em cães na região metropolitana do Rio de Janeiro. Pesquisa Veterinária Brasileira 24 (4):203-206. Depto Medicina e Cirurgia Veterinária, Inst. Veterinária, UFRRJ, Seropédica, RJ 23851-970, Brazil. E-mail: fabiolanc@ig.com.br Lyme disease is a worldwide zoonosis caused by Borrelia burgdorferi (Spirochaetae) and transmitted through ticks (Ixodidae). Many species of domestic and wild animals are affected. The dog is an important reservoir in the domiciliary environment. The disease affects the joints and causes alterations in other organs. Blood serum samples from 143 dogs, living in the counties of Seropédica, Itaguaí and Rio de Janeiro, were analyzed with the indirect ELISA test, to search for antibodies against Borrelia burgdorferi latu sensu. Average positive results were 48.25%, 41.9% in Seropédica, 4.9% in Itaguaí and 1.4% in Rio de Janeiro. The titres varied from 1:400 to 1:6400. The prevalence was lower (p< 0.05) in dogs up to one year of age than in older dogs, with significant difference. Between males and females was no statistical difference with positives titres. The result of this survey should draw attention to the occurrence of Borrelia sp in the area studied, considering the importance of Lyme disease as an emerging zoonosis.

Abstract in English:

ABSTRACT.- Marinho M., Langoni H., Oliveira S.L., Carreira R., Perri, S.H.V. e Luvizoto M.C. 2002. [Humoral iminune response, bacterial recovery and time lesions in mice geneticaly selected for high and low antibody production and in outbreed Balb/c mice face to Leptospira interrogans sorovar icterohaemorrhagiae.] Resposta humoral, recuperação bacteriana e lesões histológicas em camundongos geneticamente selecionados para bons e maus produtores de anticorpos e Balb/c, frente à infecção por Leptospira interrogans sorovar icterohaemorrhagiae. Pesquisa Veterinária Brasileira 23(1):5-12. Laboratório de Microbiologia, Departamento de Apoio, Produção e Saúde Animal FOA, Unesp-Campus de Araçatuba, Cx. Postal 341, Araçatuba, SP 16050-680, Brazil. The aim of the present work was to evaluate the association among the kinetics of humoral immune response, the recovery ofviable leptospiras and the intensity ofthe tissue lesions in mice selected for high (H) and low (L) production of antibodies (selection N-A) and in mice from the outbreed Balb/c Iine, inoculated with pathogenic Leptospira interrogans serovar icterohaemorrhagiae. Toe H and L lines showed modifications in some steps ofthe immune response, mainly in relation to the macrophagic activity, representing extreme phenotypes found in heterogeneous natural populations. Mice were sacrificed in eight moments after the infection. Analysis of the results revealed that from the 7th day after the infection, on Iine H mice presented antibodies titles significantly higher as compared to the L line mice, maintaining the multispecific effect Balb/c tine mice showed intermediàte results between the two Iines. Antibodies production worked as a timiting factor to the infection, becausewhen a greater leptospiras recovery was obtained, at the initial phase of the infection, the titles of antibodies were elevated. Inflammatory and degenerative process led to similar lesions in the organs ofinfected mice. Only a variation in the degree oftissue compromising was observed according to the tine. H tine mice showed more extensive lesions than L and Balb/c lines mice. For the Balb/c tine mice, the lesions were moderate. As a mie, H and Balb/c mice lines showed a Th2 profile of immune response, with higher indexes of antibody production and gravity ofthe lesions, while L line mice presented a Th 1 profile ofimmune response.

• Leptospirosis humoral immune response bacterial recovery lesions mice genetically selected Balb/c Leptospirose recuperação bacteriana lesões resposta humoral camundongos geneticamente selecionados.

Abstract in Portuguese:

RESUMO.- Marinho M., Langoni H., Oliveira S.L., Carreira R., Perri, S.H.V. e Luvizoto M.C. 2002. [Humoral iminune response, bacterial recovery and time lesions in mice geneticaly selected for high and low antibody production and in outbreed Balb/c mice face to Leptospira interrogans sorovar icterohaemorrhagiae.] Resposta humoral, recuperação bacteriana e lesões histológicas em camundongos geneticamente selecionados para bons e maus produtores de anticorpos e Balb/c, frente à infecção por Leptospira interrogans sorovar icterohaemorrhagiae. Pesquisa Veterinária Brasileira 23(1):5-12. Laboratório de Microbiologia, Departamento de Apoio, Produção e Saúde Animal FOA, Unesp-Campus de Araçatuba, Cx. Postal 341, Araçatuba, SP 16050-680, Brazil. O presente trabalho teve por finalidade associar a cinética da resposta humoral à recuperação de leptospiras viáveis e à intensidade das lesões teciduais em camundongos geneticamente selecionados para bons (High) e maus (Low) produtores de anticorpos (seleção IV-A), além de camundongos outbreed, Balb/c, inoculados com amostra patogênica de Leptospira interrogans sorovar icterohaemorrhagiae. As linhagens High e Low (seleção IV-A) apresentam modificações em alguns compartimentos da resposta imune, principalmente em relação à atividade macrofágica, representando fenótipos extre mos encontrados em populações naturais heterogêneas. Os camundongos foram sacrificados em oito momentos após a infecção. A análise dos resultados revelou que a partir do 7º dia após a infecção, os camundongos da linhagem High apresentaram elevação nos títulos de anticorpos estatisticamente significantes quando comparados aos camundongos da linhagem Low, mantendo assim o efeito multiespecífico. Os camundongos Balb/c apresentaram resultados intermediários entre as duas linhagens. A produção de anticorpos colaborou como fator limitante à infecção, pois quando obtevese maior recuperação de leptospiras, na fase inicial da infecção, os títulos de anticorpos encontravam-se em elevação. As lesões observadas nos órgãos de camundongos infectados consistiram basicamente nos mesmos processos inflamatórios e degenerativos, que não se alteraram, variando apenas o grau de comprometimento tecidual, de acordo com a linhagem. A linhagem high apresentou lesões mais extensas que as apresentadas pelas linhagens low e Balb/c, sendo que nesta última as lesões foram moderadas. De forma geral a linhagem High e Balb/c apresentaram um perfil de resposta Th2, com o maior índice de produção de anticorpos e gravidade das lesões, enquanto a linhagem Low apresentou um perfil de resposta Th 1.

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.