PESQUISA VETERINÁRIA BRASILEIRA

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.- Vieira R.C., Bombardiere E., Oliveira J.J., Lino-Júnior R.S., Brito L.A.B. & Junqueira-Kipnis A.P. 2008. [Influence of Copaifera langsdorffii oil on the repair of a surgical wound in the presence of foreign body.] Influência do óleo de Copaifera langsdorffii no reparo de ferida cirúrgica em presença de corpo estranho. Revista Pesquisa Veterinária Brasileira 28(8):358-366. Instituto de Patologia Tropical e Saúde Pública (IPTSP), UFG, Rua 235 s/n, Sala 325, Setor Universitário, Goiânia, GO 74605-050, Brazil. E-mail: rafael.objetivo@gmail.com Copaifera langsdorffii is a Brazilian native leguminosae that produce resin-oil, popularly known as copaíba oil. This oil is used for the treatment of skin wound due to its recognized antiinflammatory and wound healing effects. Despite, its popular use, there are few published data about the therapeutic effect of this medicinal plant. The aim of the study was to evaluate the topic treatment effect of the Copaíba oil on the process of skin repair inflammation induced by a foreign body subcutanously implanted. Sixty BALB/c mice were submitted to a 1cm linear incision and a 12mm circle coverslip was subcutaneously implanted. Four treatments groups were established: control, sterile saline (C); vehicle control, sterile mineral oil, (VC); treatment 1 (T1), mineral oil plus copaiba oil (V/V), and treatment 2 (T2) copaiba oil. The evaluations were performed at pre-determined time points (1, 3, 5, 7 and 14 days). It was possible to find fibroblasts, epithelial cells proliferation, re-epithelization and newly formed blood vessels in all groups, however, all oil treated groups (T1 and T2) did not present re-epithelization at three days post surgical incision. On days 5 and 7, a higher intensity of edema and hyperemia on the groups T1 and T2 was observed, besides that, the T1 and T2 groups presented a serous cellular scab on the wounds that was absent on the C and VC groups. The inflammatory reactions among the groups C and VC showed more mononuclear cells than the T1 and T2 groups that presented a mixed cell patter composed from both mono and polymorphonuclear cells. Although the surgical wounds were re-epithelizaded, in the groups T1 and T2, they were covered by a serous cellular crust and the dermis tissue still presented an intense mononuclear cell inflammatory focus. Fourteen days after of the surgical incision, the gross aspects on groups C and VC were similar and on groups T1 and T2, despite wound to be completely closed and without crusts, the skin those animals was thickened. Furthermore, the dermis on group T2 presented moderate fibrosis, while the other groups presented slightly ones. The results demonstrated that topical treatment with C. langsdorffii oil debilitated the normal process of a wound repair in the presence of a foreign body.

• Copaifera langsdorffii copaíba wound healing anti-inflammatory

Abstract in Portuguese:

ABSTRACT.- Vieira R.C., Bombardiere E., Oliveira J.J., Lino-Júnior R.S., Brito L.A.B. & Junqueira-Kipnis A.P. 2008. [Influence of Copaifera langsdorffii oil on the repair of a surgical wound in the presence of foreign body.] Influência do óleo de Copaifera langsdorffii no reparo de ferida cirúrgica em presença de corpo estranho. Revista Pesquisa Veterinária Brasileira 28(8):358-366. Instituto de Patologia Tropical e Saúde Pública (IPTSP), UFG, Rua 235 s/n, Sala 325, Setor Universitário, Goiânia, GO 74605-050, Brazil. E-mail: rafael.objetivo@gmail.com Copaifera langsdorffii is a Brazilian native leguminosae that produce resin-oil, popularly known as copaíba oil. This oil is used for the treatment of skin wound due to its recognized antiinflammatory and wound healing effects. Despite, its popular use, there are few published data about the therapeutic effect of this medicinal plant. The aim of the study was to evaluate the topic treatment effect of the Copaíba oil on the process of skin repair inflammation induced by a foreign body subcutanously implanted. Sixty BALB/c mice were submitted to a 1cm linear incision and a 12mm circle coverslip was subcutaneously implanted. Four treatments groups were established: control, sterile saline (C); vehicle control, sterile mineral oil, (VC); treatment 1 (T1), mineral oil plus copaiba oil (V/V), and treatment 2 (T2) copaiba oil. The evaluations were performed at pre-determined time points (1, 3, 5, 7 and 14 days). It was possible to find fibroblasts, epithelial cells proliferation, re-epithelization and newly formed blood vessels in all groups, however, all oil treated groups (T1 and T2) did not present re-epithelization at three days post surgical incision. On days 5 and 7, a higher intensity of edema and hyperemia on the groups T1 and T2 was observed, besides that, the T1 and T2 groups presented a serous cellular scab on the wounds that was absent on the C and VC groups. The inflammatory reactions among the groups C and VC showed more mononuclear cells than the T1 and T2 groups that presented a mixed cell patter composed from both mono and polymorphonuclear cells. Although the surgical wounds were re-epithelizaded, in the groups T1 and T2, they were covered by a serous cellular crust and the dermis tissue still presented an intense mononuclear cell inflammatory focus. Fourteen days after of the surgical incision, the gross aspects on groups C and VC were similar and on groups T1 and T2, despite wound to be completely closed and without crusts, the skin those animals was thickened. Furthermore, the dermis on group T2 presented moderate fibrosis, while the other groups presented slightly ones. The results demonstrated that topical treatment with C. langsdorffii oil debilitated the normal process of a wound repair in the presence of a foreign body.

Abstract in English:

ABSTRACT.- La Paz M.N., Fonseca V.U., Campos D.B., Artoni L.P., Sousa L.M.M.C. & Papa P.C. 2007. [In vitro progesterone production from bovine corpus luteum throughout gestation.] Produção de progesterona in vitro pelas células do corpo lúteo bovino ao longo da gestação. Pesquisa Veterinária Brasileira 27(9):370376. Setor de Anatomia, Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques Paiva, 87, SP 05508-270, Brazil. E-mail: ppapa@usp.br The aim was to test the hypothesis that cultivated bovine luteal cells from three different thirds of pregnancy behave the same way as in vivo luteal cells relative to P4 production. Corpus luteum samples from days 90 (n=3), 150 (n=3) and 210 (n=3) of pregnancy were obtained at a local slaughterhouse. Under aseptic conditions cells were mechanically dispersed and cultivated in a 96 wells-plate. After 24 hours of culture, cells were washed and the precursor pregnenolone was added. Experiments were conducted eight times for each studied time period (24, 48 and 96 h) and three times for each gestational age. Culture medium and cells were collected after 24, 48 and 96 hours of precursor addition and kept frozen at -20oC until processing. Progesterone was measured by RIA and protein content by Lowry’s method. Results were statistically analyzed and considered different when p <0.05. A higher P4 production was observed on day 90 of gestation (35.277±0.075), then this production was decreased at day 150 (28.820±0.231) and increased again at day 210 (32.777±0.099). After 24 hours of culture, luteal cells P4 production reached maximum values in the group of 90 days (2.912±0.047) when compared to 150 (2.669±0.137) and 210 days (2.741±0.088). At 48 and 96 hours of culture, bovine luteal cells from day 90 of gestation produced more P4 than cells from day 210 (2.934±0.029 and 2.976±0.121 respectively x 2.760±0.059 and 2.695±0.149, respectively; p<0.05), which in turn, produced more P4 than cells from day 150 (2.334±0.084 for 48 h and 2.205±0.136 for 96 h). Luteal cells from day 150 of gestation presented a decreasing P4 production throughout the 96 hours of culture. These differences could be explained by differential gene expression of enzymes and/or factors belonging to the esteroidogenic cascade in accordance to the gestational period. The established luteal cell culture model could be used for further functional studies once P4 secretion pattern in vitro resembled what occurs in vivo.

• Corpus luteum gestation progesterone cell culture

Abstract in Portuguese:

ABSTRACT.- La Paz M.N., Fonseca V.U., Campos D.B., Artoni L.P., Sousa L.M.M.C. & Papa P.C. 2007. [In vitro progesterone production from bovine corpus luteum throughout gestation.] Produção de progesterona in vitro pelas células do corpo lúteo bovino ao longo da gestação. Pesquisa Veterinária Brasileira 27(9):370376. Setor de Anatomia, Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques Paiva, 87, SP 05508-270, Brazil. E-mail: ppapa@usp.br The aim was to test the hypothesis that cultivated bovine luteal cells from three different thirds of pregnancy behave the same way as in vivo luteal cells relative to P4 production. Corpus luteum samples from days 90 (n=3), 150 (n=3) and 210 (n=3) of pregnancy were obtained at a local slaughterhouse. Under aseptic conditions cells were mechanically dispersed and cultivated in a 96 wells-plate. After 24 hours of culture, cells were washed and the precursor pregnenolone was added. Experiments were conducted eight times for each studied time period (24, 48 and 96 h) and three times for each gestational age. Culture medium and cells were collected after 24, 48 and 96 hours of precursor addition and kept frozen at -20oC until processing. Progesterone was measured by RIA and protein content by Lowry’s method. Results were statistically analyzed and considered different when p <0.05. A higher P4 production was observed on day 90 of gestation (35.277±0.075), then this production was decreased at day 150 (28.820±0.231) and increased again at day 210 (32.777±0.099). After 24 hours of culture, luteal cells P4 production reached maximum values in the group of 90 days (2.912±0.047) when compared to 150 (2.669±0.137) and 210 days (2.741±0.088). At 48 and 96 hours of culture, bovine luteal cells from day 90 of gestation produced more P4 than cells from day 210 (2.934±0.029 and 2.976±0.121 respectively x 2.760±0.059 and 2.695±0.149, respectively; p<0.05), which in turn, produced more P4 than cells from day 150 (2.334±0.084 for 48 h and 2.205±0.136 for 96 h). Luteal cells from day 150 of gestation presented a decreasing P4 production throughout the 96 hours of culture. These differences could be explained by differential gene expression of enzymes and/or factors belonging to the esteroidogenic cascade in accordance to the gestational period. The established luteal cell culture model could be used for further functional studies once P4 secretion pattern in vitro resembled what occurs in vivo.

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.- Oliveira C.M.C., Barbosa J.D., Pfeifer I.B. & Cardoso D.P. 2003. [The blood and urinary values in the pre and post-parturient period of buffaloes, kept on exclusive pasture feeding.] Parâmetros sangüineos e urinários, no pré e pós-parto, de búfalas criadas em sistema exclusivo de pastejo. Pesquisa Veterinária Brasileira 23(2):87-92. Central de Diagnóstico da Universidade Federal do Pará, Rua Maximino Porpino 1000, Castanhal, PA 68740-080, Brazil. E-mail: diomedes@ufpa.br Eight lactating buffaloes of the Murrah breed were used from 60 days before up to 60 days after calving, in order to evaluate alterations in the concentration of blood glucose, total plasmatic protein, haematocrit and the presence of ketonic bodies in the urine. The 5 to 10-year old buffaloes were clinically healthy and had had at least two lactations. During the experimental period the animais were maintained on Brachiaria brizantha pasture and · supplemented with a mineral mixture and water. The media concentrations of blood glucose and the media values of the haematocrit diminished significantly after calving (p<0,05). The concentrations of total plasmatic protein did not show significant variation during the pre and post-parturient period. The ketonic bodies in the pre-parturient period were only detected in the urine of one buffaloe, but after the 32nd day of lactation ketonic bodies were detected in ali animais. There was a direct relationship between the color of the urine positive by the Rothera test and blood glucose concentrations. lt can be concluded that at the beginning of lactation the buffaloes had an energetic deficit, characterized by a decline of blood glucose concentrations and the presence of ketonic bodies in the urine, and that lactation caused a progressive decline of the hematocrit, but that the concentration of total plasmatic protein did not vary during the pre and post-parturient period.

• Buffaloes ketonic bodies lactation metabolic perfil urine Búfalas corpos cetônicos lactação perfil metabólico urina.

Abstract in Portuguese:

RESUMO.- Oliveira C.M.C., Barbosa J.D., Pfeifer I.B. & Cardoso D.P. 2003. [The blood and urinary values in the pre and post-parturient period of buffaloes, kept on exclusive pasture feeding.] Parâmetros sangüineos e urinários, no pré e pós-parto, de búfalas criadas em sistema exclusivo de pastejo. Pesquisa Veterinária Brasileira 23(2):87-92. Central de Diagnóstico da Universidade Federal do Pará, Rua Maximino Porpino 1000, Castanhal, PA 68740-080, Brazil. E-mail: diomedes@ufpa.br Foi avaliada a ocorrência de alterações nas concentrações de glicose sangüínea, proteína plasmática total; hematócrito e presença de corpos cetônicos na urina de oito búfalas leiteiras da raça Murrah, com idade variando entre 5 e 10 anos, com no mínimo duas lactações, clinicamente sadias, desde 60 dias antes até 60 dias pós-parto. As concentrações médias de glicose sangüínea e o valor médio do hematócrito diminuíram significativamente no pós-parto (p<0,05). As concentrações de proteína plasmática total não sofreram variações significativas do pré para o pós-parto. No período préparto os corpos cetônicos só foram detectados na urina de uma búfala; entretanto, a partir do 32º dia de lactação foram detectados em todos os animais. Houve uma relação direta entre a coloração da urina positiva para o teste de Rothera e as concentrações de glicose sangüínea. Pode-se concluir que na fase inicial da lactação as búfalas utilizadas sofreram um déficit energético, caracterizado pela diminuição nas concentrações sangüíneas de glicose e presença de corpos cetônicos na urina, e que a lactação causou um declínio progressivo no hematócrito, enquanto que as concentrações da proteína plasmática total não sofreram variações do pré para o pós-parto.

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.