PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- Scarpelli L., Lopes W.D.Z., Migani M., Bresciani K.D.S. & Costa A.J. 2009. Toxoplasma gondii in experimentally infected Bos taurus and Bos indicus semen and tissues. Pesquisa Veterinária Brasileira 29(1):59-64. Departamento de Medicina Veterinária Preventiva, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Via de Acesso Prof. Paulo Donatto Castellani s/n, Jaboticabal, SP 14884-900, Brazil. E-mail: wdzlopes@hotmail.com Eighteen young steers were inoculated with Toxoplasma gondii and randomly distributed into three groups of six animals each: GI, 2.5x105 “P” strain oocysts, GII, 5.0x106 “RH” strain tachyzoites, and GIII (Control). Clinical, serological and parasitemia exams were realized. Parasite investigation by bioassay and PCR was realized on semen and fragments of skeletal musculature, lymph nodes, brain, retina, spleen, liver, lung, testicle, epididymis and seminal vesicle. Blood and semen samples were collected on days -2, -1, 1, 3, 5, 7, 14 and weekly thereafter, up to postinfection day (PID) 84. The inoculated steers (GI and GII) presented hyperthermia from PID 3 to 16. Antibodies against T. gondii were detected through the indirect fluorescence antibody test (IFAT) on PID 5 (1:16) in both inoculated groups (oocysts and tachyzoites), reaching peaks of 1:4096 on PID 7. Parasitemia outbursts occurred in all infected bovines, principally from PID 7 to 28, independent of the strain and inoculate used. Bioassays revealed the presence of parasites in semen samples of animals infected with oocysts (GI) and tachyzoites (GII) on several experimental days between PID 7 and 84. Tissue parasitism by T. gondii was diagnosed by bioassay and the PCR technique in several organ and tissue fragments. These findings suggest the possibility of sexual transmission of T. gondii in the bovine species.

• Cattle PCR semen Toxoplasma gondii Apicomplexa

Abstract in Portuguese:

ABSTRACT.- Scarpelli L., Lopes W.D.Z., Migani M., Bresciani K.D.S. & Costa A.J. 2009. Toxoplasma gondii in experimentally infected Bos taurus and Bos indicus semen and tissues. Pesquisa Veterinária Brasileira 29(1):59-64. Departamento de Medicina Veterinária Preventiva, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Via de Acesso Prof. Paulo Donatto Castellani s/n, Jaboticabal, SP 14884-900, Brazil. E-mail: wdzlopes@hotmail.com Eighteen young steers were inoculated with Toxoplasma gondii and randomly distributed into three groups of six animals each: GI, 2.5x105 “P” strain oocysts, GII, 5.0x106 “RH” strain tachyzoites, and GIII (Control). Clinical, serological and parasitemia exams were realized. Parasite investigation by bioassay and PCR was realized on semen and fragments of skeletal musculature, lymph nodes, brain, retina, spleen, liver, lung, testicle, epididymis and seminal vesicle. Blood and semen samples were collected on days -2, -1, 1, 3, 5, 7, 14 and weekly thereafter, up to postinfection day (PID) 84. The inoculated steers (GI and GII) presented hyperthermia from PID 3 to 16. Antibodies against T. gondii were detected through the indirect fluorescence antibody test (IFAT) on PID 5 (1:16) in both inoculated groups (oocysts and tachyzoites), reaching peaks of 1:4096 on PID 7. Parasitemia outbursts occurred in all infected bovines, principally from PID 7 to 28, independent of the strain and inoculate used. Bioassays revealed the presence of parasites in semen samples of animals infected with oocysts (GI) and tachyzoites (GII) on several experimental days between PID 7 and 84. Tissue parasitism by T. gondii was diagnosed by bioassay and the PCR technique in several organ and tissue fragments. These findings suggest the possibility of sexual transmission of T. gondii in the bovine species.

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.- Costa M.C., Flaiban K.K.M.C., Coneglian M.M., Dognani R., Vettorato E.D., Balarin M.R.S. & Lisbôa J.A.N. 2008. [Neutrophil oxidative burst in Nelore and Limousin calves in the first four months of life.] Metabolismo oxidativo dos neutrófilos de bezerros das raças Nelore e Limousin nos primeiros quatro meses de vida. Pesquisa Veterinária Brasileira 28(9):431-436. Departamento de Clínicas Veterinárias, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Campus Universitário, Cx. Postal 6001, Londrina, PR 86051-990, Brazil. E-mail: janlisboa@uel.br To study some defence mechanisms of beef calves kept on range conditions, 90 healthy calves, 45 Nelore and 45 Limousin, were bled at 24 to 36 hours of life, 15, 30, 60, 90 and, 120 days. Leukogram and neutrophil oxidative burst through the non-stimulated and the stimulated NBT tests were determined. The parasitemia with Anaplasma marginale, Babesia bigemina and B. bovis was investigated. Repeated measure ANOVA was used to analyse the age effect on leukocyte counts. Variation of positive neutrophil throughout the age was analysed through Krushkal-Wallis test. Comparisons among breeds were done using the Mann-Whithney test. The white blood cell count increased with age in both breeds, with a decrease in neutrophil and an increase in lymphocyte counts. The neutrophil lymphocyte ratio had been inverted before 15 days of life. The neutrophil oxidative burst was less effective in the newborn calves increasing with age. Limousin calves’ neutrophils had higher capacity to reduce NBT in advanced ages. The observed differences between taurine and Zebu calves had no effects on calves’ health and could be atributed to Anaplasma marginale natural and asynptomatic infection.

• Beef calf Zebu taurine NBT Anaplasma marginale

Abstract in Portuguese:

ABSTRACT.- Costa M.C., Flaiban K.K.M.C., Coneglian M.M., Dognani R., Vettorato E.D., Balarin M.R.S. & Lisbôa J.A.N. 2008. [Neutrophil oxidative burst in Nelore and Limousin calves in the first four months of life.] Metabolismo oxidativo dos neutrófilos de bezerros das raças Nelore e Limousin nos primeiros quatro meses de vida. Pesquisa Veterinária Brasileira 28(9):431-436. Departamento de Clínicas Veterinárias, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Campus Universitário, Cx. Postal 6001, Londrina, PR 86051-990, Brazil. E-mail: janlisboa@uel.br To study some defence mechanisms of beef calves kept on range conditions, 90 healthy calves, 45 Nelore and 45 Limousin, were bled at 24 to 36 hours of life, 15, 30, 60, 90 and, 120 days. Leukogram and neutrophil oxidative burst through the non-stimulated and the stimulated NBT tests were determined. The parasitemia with Anaplasma marginale, Babesia bigemina and B. bovis was investigated. Repeated measure ANOVA was used to analyse the age effect on leukocyte counts. Variation of positive neutrophil throughout the age was analysed through Krushkal-Wallis test. Comparisons among breeds were done using the Mann-Whithney test. The white blood cell count increased with age in both breeds, with a decrease in neutrophil and an increase in lymphocyte counts. The neutrophil lymphocyte ratio had been inverted before 15 days of life. The neutrophil oxidative burst was less effective in the newborn calves increasing with age. Limousin calves’ neutrophils had higher capacity to reduce NBT in advanced ages. The observed differences between taurine and Zebu calves had no effects on calves’ health and could be atributed to Anaplasma marginale natural and asynptomatic infection.

Abstract in English:

ABSTRACT.- Botteon P.T.L., Ferraz V.P., Glória M.B.A., Melo M.M., Paes Leme F.O., Faleiros R.R. & Alves G.E.S. 2008. [Plasmatic concentrations of tryptamine, tyramine end phenylethylamine in horses under the effect of carbohydrate overload and non-steroid antinflammatory compounds.] Concentrações plasmáticas de triptamina, tiramina e feniletilamina em eqüinos sob efeitos de sobrecarga de carboidratos e antiinflamatórios não esteroidais. Pesquisa Veterinária Brasileira 28(6):299-302. Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Pampulha, Belo Horizonte, MG, 30123-970, Brazil. E-mail: paulobotteon@gmail.com The concentrations of the bioactives amines tryptamine (TRP), tyramine (TYR) and phenylethylamine (PEA) were determined by gas chromatography in plasma samples of 20 horses submitted to carbohydrate overload. Thirty hours after the overload, the horses were randomly distributed in four groups (n=5) and were submitted to four IV treatments every 12 hours: 10ml of saline (GC), ketoprofen 2.2mg/kg (GK), phenylbutazone 4.4mg/kg (GF), and flunixin meglumine 1.1mg/kg (GFM). Blood samples were collected at various times after the overload (0-72 h). Plasma TYR and PEA concentrations ranged from 0.18 to 164.2mg/L, and differed significantly with time (p<0.01), but did not differ in the treatments. Plasma concentrations of TRP differed between times and treatments. The GC was significantly major than other treatments at 48h and 60h after the overload, and the plasma concentration of TRP in groups GK and GFM was significantly lower than in groups GF and GC at 72 h (p=0.0012). We concluded that the anti-inflammatory drugs evaluated do not interfere in the plasma concentration of TYP and PEA. For TRP, ketoprofen and flunixin meglumine was effective to reduce de plasmatic concentration of this amine.

• Amines antinflammatory compounds laminitis horse

Abstract in Portuguese:

ABSTRACT.- Botteon P.T.L., Ferraz V.P., Glória M.B.A., Melo M.M., Paes Leme F.O., Faleiros R.R. & Alves G.E.S. 2008. [Plasmatic concentrations of tryptamine, tyramine end phenylethylamine in horses under the effect of carbohydrate overload and non-steroid antinflammatory compounds.] Concentrações plasmáticas de triptamina, tiramina e feniletilamina em eqüinos sob efeitos de sobrecarga de carboidratos e antiinflamatórios não esteroidais. Pesquisa Veterinária Brasileira 28(6):299-302. Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais, Av. Antônio Carlos 6627, Pampulha, Belo Horizonte, MG, 30123-970, Brazil. E-mail: paulobotteon@gmail.com The concentrations of the bioactives amines tryptamine (TRP), tyramine (TYR) and phenylethylamine (PEA) were determined by gas chromatography in plasma samples of 20 horses submitted to carbohydrate overload. Thirty hours after the overload, the horses were randomly distributed in four groups (n=5) and were submitted to four IV treatments every 12 hours: 10ml of saline (GC), ketoprofen 2.2mg/kg (GK), phenylbutazone 4.4mg/kg (GF), and flunixin meglumine 1.1mg/kg (GFM). Blood samples were collected at various times after the overload (0-72 h). Plasma TYR and PEA concentrations ranged from 0.18 to 164.2mg/L, and differed significantly with time (p<0.01), but did not differ in the treatments. Plasma concentrations of TRP differed between times and treatments. The GC was significantly major than other treatments at 48h and 60h after the overload, and the plasma concentration of TRP in groups GK and GFM was significantly lower than in groups GF and GC at 72 h (p=0.0012). We concluded that the anti-inflammatory drugs evaluated do not interfere in the plasma concentration of TYP and PEA. For TRP, ketoprofen and flunixin meglumine was effective to reduce de plasmatic concentration of this amine.

Abstract in English:

ABSTRACT.- Zuccari D.A.P.C., Pavam M.V., Terzian A.C.B., Pereira R.S., Ruiz C.M. & Andrade J.C.† 2008. Immunohistochemical evaluation of Ki-67 and PCNA in canine mammary neoplasias: Correlation with prognostic factors and clinical outcome. Pesquisa Veterinária Brasileira 28(4):207-215. Centro Regional de Bioterismo, FAMERP, Av. Brigadeiro Faria Lima 5416, São José do Rio Preto, SP 15090-000, Brazil. E-mail: debora.zuccari@famerp.br E-cadherin is a cell-cell adhesion molecule and low e-cadherin expression is related to invasiveness and may indicate a bad prognosis in mammary neoplasms. The expression of cell proliferation markers PCNA and especially Ki-67, has also proved to have a strong prognostic value in this tumor class. The expression of these markers was related to the clinical-pathological characteristics of 73 surgically removed mammary tumors in female dogs by immunohistochemistry. There was no statistical correlation between these markers and death by neoplasm, survival time and disease-free interval. However, the loss of e-cadherin expression and marked Ki-67 expression (p=0.016) were considered statistically significant for the diagnosis (p=0.032). When evaluated as independent factors, there was evidence of the relationship between the loss of e-cadherin expression and high PCNA expression with changes in the body status (divided into obese, normal and cachectic) of female dogs (p=0.030); there was also evidence of the relationship between pseudopregnancy and e-cadherin alone (p=0.021) and for ulceration and PCNA alone (p=0.035). The significant correlation between the markers expression and these well known prognostic factors used individually or in combination suggests their prognostic value in canine mammary tumors.

• Canine cell proliferation marker e-cadherin immunohistochemistry mammary neoplasm

Abstract in Portuguese:

ABSTRACT.- Zuccari D.A.P.C., Pavam M.V., Terzian A.C.B., Pereira R.S., Ruiz C.M. & Andrade J.C.† 2008. Immunohistochemical evaluation of Ki-67 and PCNA in canine mammary neoplasias: Correlation with prognostic factors and clinical outcome. Pesquisa Veterinária Brasileira 28(4):207-215. Centro Regional de Bioterismo, FAMERP, Av. Brigadeiro Faria Lima 5416, São José do Rio Preto, SP 15090-000, Brazil. E-mail: debora.zuccari@famerp.br E-cadherin is a cell-cell adhesion molecule and low e-cadherin expression is related to invasiveness and may indicate a bad prognosis in mammary neoplasms. The expression of cell proliferation markers PCNA and especially Ki-67, has also proved to have a strong prognostic value in this tumor class. The expression of these markers was related to the clinical-pathological characteristics of 73 surgically removed mammary tumors in female dogs by immunohistochemistry. There was no statistical correlation between these markers and death by neoplasm, survival time and disease-free interval. However, the loss of e-cadherin expression and marked Ki-67 expression (p=0.016) were considered statistically significant for the diagnosis (p=0.032). When evaluated as independent factors, there was evidence of the relationship between the loss of e-cadherin expression and high PCNA expression with changes in the body status (divided into obese, normal and cachectic) of female dogs (p=0.030); there was also evidence of the relationship between pseudopregnancy and e-cadherin alone (p=0.021) and for ulceration and PCNA alone (p=0.035). The significant correlation between the markers expression and these well known prognostic factors used individually or in combination suggests their prognostic value in canine mammary tumors.

Abstract in English:

ABSTRACT.- Millar P.R., Daguer H., Vicente R.T., Costa T., Sobreiro L.G. & Amendoeira M.R.R. 2008. [Toxoplasma gondii: epidemiological study of pigs from southwestern Paraná, Brazil.] Toxoplasma gondii: estudo soro-epidemiológico de suínos da região Sudoeste do Estado do Paraná. Pesquisa Veterinária Brasileira 28(1):15-18. Programa de Pós-Graduação em Higiene Veterinária e Processamento Tecnológico de Produtos de Origem Animal, Universidade Federal Fluminense, Rua Vital Brazil Filho 64, Santa Rosa, Niterói, RJ 24230-360, Brazil. E-mail: patriciariddell@vm.uff.br This study evaluated the seroprevalence of toxoplasmosis in pigs bred and slaughtered for human consumption. The frequency of the antibody anti-Toxoplasma gondii was investigated in 408 animals from distinct piggeries of 25 cities in the southwestern region of the state of Paraná, Brazil. Serum samples were examined for IgG antibodies specific for T. gondii through the indirect fluorescent antibody test (IFAT). A frequency of 25.5% of serum samples tested positive for dilutions equal to or greater than 1:64. Statistically significant were not observed in relation to sex and age of the animals. The highest antibody level observed was 4,096 (3.0%) and the most frequent was 64 (74%), suggesting that a chronic infection occurred in the population. The high prevalence of the infection found in the swine herds of this region suggests that these herds can be a source of infection for individuals who typically consume this type of raw or badly cooked meat.

• Toxoplasma gondii pigs seroprevalence

Abstract in Portuguese:

ABSTRACT.- Millar P.R., Daguer H., Vicente R.T., Costa T., Sobreiro L.G. & Amendoeira M.R.R. 2008. [Toxoplasma gondii: epidemiological study of pigs from southwestern Paraná, Brazil.] Toxoplasma gondii: estudo soro-epidemiológico de suínos da região Sudoeste do Estado do Paraná. Pesquisa Veterinária Brasileira 28(1):15-18. Programa de Pós-Graduação em Higiene Veterinária e Processamento Tecnológico de Produtos de Origem Animal, Universidade Federal Fluminense, Rua Vital Brazil Filho 64, Santa Rosa, Niterói, RJ 24230-360, Brazil. E-mail: patriciariddell@vm.uff.br This study evaluated the seroprevalence of toxoplasmosis in pigs bred and slaughtered for human consumption. The frequency of the antibody anti-Toxoplasma gondii was investigated in 408 animals from distinct piggeries of 25 cities in the southwestern region of the state of Paraná, Brazil. Serum samples were examined for IgG antibodies specific for T. gondii through the indirect fluorescent antibody test (IFAT). A frequency of 25.5% of serum samples tested positive for dilutions equal to or greater than 1:64. Statistically significant were not observed in relation to sex and age of the animals. The highest antibody level observed was 4,096 (3.0%) and the most frequent was 64 (74%), suggesting that a chronic infection occurred in the population. The high prevalence of the infection found in the swine herds of this region suggests that these herds can be a source of infection for individuals who typically consume this type of raw or badly cooked meat.

Abstract in English:

ABSTRACT.- Watanabe I., Guimarães J.P., Ogawa K., Iyomasa M.M., Miglino M.A. ,Silva M. C.P., Semprini M., Sosthines M.C.K., Lopes M.O. & Lopes R.A. 2007. [Submandibular gland of rats with ageing: observations with high resolution scanning electron microscopy.] Glândula submandibular de ratos com envelhecimento: observações ao microscópio eletrônico de varredura de alta resolução. Pesquisa Veterinária Brasileira 27(12):501-505. Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Cidade Universitária, Av. Prof. Dr. Orlando Maruer de Paiva 87, 05389-970, São Paulo, SP, Brazil. E-mail: watanabe@icb.usp.br The three-dimensional characteristics of the intracellular components of acinar and ductal cells were revealed using the osmium-DMSO-osmium method. The samples were macerated in diluted osmium after fractured in DMSO solution. The stacks of the rough endoplasmic reticulum are revealed intermingling by several mitochondria. The lamellae of the rough endoplasmic reticulum are located around the nuclei at basal portion and these structures are shown in three-dimensional HRSEM images.

• Submandibular gland rough endoplasmic reticulum mitochondria rat HRSEM

Abstract in Portuguese:

ABSTRACT.- Watanabe I., Guimarães J.P., Ogawa K., Iyomasa M.M., Miglino M.A. ,Silva M. C.P., Semprini M., Sosthines M.C.K., Lopes M.O. & Lopes R.A. 2007. [Submandibular gland of rats with ageing: observations with high resolution scanning electron microscopy.] Glândula submandibular de ratos com envelhecimento: observações ao microscópio eletrônico de varredura de alta resolução. Pesquisa Veterinária Brasileira 27(12):501-505. Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Cidade Universitária, Av. Prof. Dr. Orlando Maruer de Paiva 87, 05389-970, São Paulo, SP, Brazil. E-mail: watanabe@icb.usp.br The three-dimensional characteristics of the intracellular components of acinar and ductal cells were revealed using the osmium-DMSO-osmium method. The samples were macerated in diluted osmium after fractured in DMSO solution. The stacks of the rough endoplasmic reticulum are revealed intermingling by several mitochondria. The lamellae of the rough endoplasmic reticulum are located around the nuclei at basal portion and these structures are shown in three-dimensional HRSEM images.

Abstract in English:

ABSTRACT.- Eigenheer-Moreira J.F., Fernandes F.T., Queiroz F.J.R, Pinho T.G.& Ferreira A.M.R. 2007. [Comparative study of repeat breeds and healthy mares through endometrial histology and plasmatic progesterone concentrations.] Estudo comparativo de éguas repetidoras ou não de cio através da avaliação histológica do endométrio e das concentrações plasmáticas de progesterona. Pesquisa Veterinária Brasileira 27(12):506-512. Curso de Pós-Graduação em Clínica e Reprodução Animal, Rua Vital Brazil Filho 64, Niterói, RJ 24230-340, Brazil. E-mail: joana.vet@gmail.com.br The study aimed to compare endometrial histology and plasmatic progesterone (P4) concentration of repeat breeds and healthy mares. The hypothesis was that there is a correlation between infertility and endometrial histology and P4 concentration in both groups. A total of 36 Campolina and Mangalarga Marchador mares in reproductive age (3-23 years) were used, 11 of them were healthy mares (Control group, 7 embryo recipient and 4 embryo donors), and 25 repeat breeders (10 embryo recipient and 15 embryo donors), classified as based on their reproductive history. Endometrial and blood samples were collected for respectively histological and plasma progesterone concentration evaluation. The endometrial samples obtained after biopsy were fixed in Bouin’s fluid, processed, included in paraffin, and stained with Hematoxylin-Eosin (HE) for histopathological examination. Plasmatic progesterone concentrations were evaluated by enzyme immunoessay (ELISA). There was no correlation between progesterone concentration and fertility. But there was a positive correlation between age and fertility, as older mares had major tendency of subfertility than younger ones. There was also a correlation between biopsy categories and fertility, as more histological alterations were found, higher were the chances for the mares to be subfertile. However not all mares classified as Category I and II maintained pregnancy until parturition. Other factors could influence pregnancy maintenance. In the same way, not all mares in Category III were infertile. The endometrial biopsy was shown to be an easy and cheap diagnostic technique with minimal discomfort to the animals and, together with other data, to be a very important component in the investigation of mare fertility.

• Mare subfertility endometrial biopsy histopathology progesterone

Abstract in Portuguese:

ABSTRACT.- Eigenheer-Moreira J.F., Fernandes F.T., Queiroz F.J.R, Pinho T.G.& Ferreira A.M.R. 2007. [Comparative study of repeat breeds and healthy mares through endometrial histology and plasmatic progesterone concentrations.] Estudo comparativo de éguas repetidoras ou não de cio através da avaliação histológica do endométrio e das concentrações plasmáticas de progesterona. Pesquisa Veterinária Brasileira 27(12):506-512. Curso de Pós-Graduação em Clínica e Reprodução Animal, Rua Vital Brazil Filho 64, Niterói, RJ 24230-340, Brazil. E-mail: joana.vet@gmail.com.br The study aimed to compare endometrial histology and plasmatic progesterone (P4) concentration of repeat breeds and healthy mares. The hypothesis was that there is a correlation between infertility and endometrial histology and P4 concentration in both groups. A total of 36 Campolina and Mangalarga Marchador mares in reproductive age (3-23 years) were used, 11 of them were healthy mares (Control group, 7 embryo recipient and 4 embryo donors), and 25 repeat breeders (10 embryo recipient and 15 embryo donors), classified as based on their reproductive history. Endometrial and blood samples were collected for respectively histological and plasma progesterone concentration evaluation. The endometrial samples obtained after biopsy were fixed in Bouin’s fluid, processed, included in paraffin, and stained with Hematoxylin-Eosin (HE) for histopathological examination. Plasmatic progesterone concentrations were evaluated by enzyme immunoessay (ELISA). There was no correlation between progesterone concentration and fertility. But there was a positive correlation between age and fertility, as older mares had major tendency of subfertility than younger ones. There was also a correlation between biopsy categories and fertility, as more histological alterations were found, higher were the chances for the mares to be subfertile. However not all mares classified as Category I and II maintained pregnancy until parturition. Other factors could influence pregnancy maintenance. In the same way, not all mares in Category III were infertile. The endometrial biopsy was shown to be an easy and cheap diagnostic technique with minimal discomfort to the animals and, together with other data, to be a very important component in the investigation of mare fertility.

Abstract in English:

ABSTRACT.- Moura A.B., Costa A.J., Jordão Filho S., Paim B.B., Pinto F.R. & Di Mauro D.C. 2007. Toxoplasma gondii in semen of experimentally infected swine. Pesquisa Veterinária Brasileira 27(10):430-434. Centro de Pesquisas em Sanidade Animal, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Jaboticabal, SP 14884-900, Brazil. E-mail: a2abm@cav.udesc.br Eight reproductive boars were divided into three groups and inoculated with Toxoplasma gondii [GI (n=3) 1.5x104 oocysts strain P; GII (n=3) 1.0x106 tachyzoites strain RH; and GIII (n=2) non-inoculated control]. Clinical, hematological, parasitemia and serological tests and studies of the parasite in the semen through bioassay and PCR, and in reproductive organs (Bioassay and immunohistochemical analyses) were conducted to evaluate the toxoplasmic infection. Blood and semen were collected on day -2, -1, 1, 3, 5, 7, 9, 11, 14 and weekly up to 84 days post-inoculation (DPI). No clinical or hematimetric alteration was observed in the boars. Parasitemia was detected in one boar inoculated with oocysts at the 7th DPI and in another boar infected with tachyzoites (GII) at the 3rd and 49th DPI. Serological tests revealed antibodies against T. gondii in animals inoculated with oocysts or tachyzoites at the 7th DPI with dilutions of 1:256 and 1:64, which reached peaks of 1:4096 at day 11 and 9, respectively. The bioassays revealed the presence of the parasite in semen samples of a boar inoculated with oocysts (GI) at 3, 49 and 56 DPI and from two boars infected with tachyzoites (GII), one animal at 5 and two animals at 49 days DPI. Mice inoculated with semen from the control group (GIII) remained serologically negative. PCR analysis showed T. gondii DNA in the semen of Boar 1 and Boar 3 inoculated with tachyzoites and oocysts, respectively. The immuno-histochemical tests showed T. gondii in the reproductive organs of Boar 1 and Boar 2, inoculated with tachyzoites and oocysts, respectively. These findings suggest the possible occurrence of venereal transmission of T. gondii in swine.

• Toxoplasmosis swine semen PCR bioassays

Abstract in Portuguese:

ABSTRACT.- Moura A.B., Costa A.J., Jordão Filho S., Paim B.B., Pinto F.R. & Di Mauro D.C. 2007. Toxoplasma gondii in semen of experimentally infected swine. Pesquisa Veterinária Brasileira 27(10):430-434. Centro de Pesquisas em Sanidade Animal, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Jaboticabal, SP 14884-900, Brazil. E-mail: a2abm@cav.udesc.br Eight reproductive boars were divided into three groups and inoculated with Toxoplasma gondii [GI (n=3) 1.5x104 oocysts strain P; GII (n=3) 1.0x106 tachyzoites strain RH; and GIII (n=2) non-inoculated control]. Clinical, hematological, parasitemia and serological tests and studies of the parasite in the semen through bioassay and PCR, and in reproductive organs (Bioassay and immunohistochemical analyses) were conducted to evaluate the toxoplasmic infection. Blood and semen were collected on day -2, -1, 1, 3, 5, 7, 9, 11, 14 and weekly up to 84 days post-inoculation (DPI). No clinical or hematimetric alteration was observed in the boars. Parasitemia was detected in one boar inoculated with oocysts at the 7th DPI and in another boar infected with tachyzoites (GII) at the 3rd and 49th DPI. Serological tests revealed antibodies against T. gondii in animals inoculated with oocysts or tachyzoites at the 7th DPI with dilutions of 1:256 and 1:64, which reached peaks of 1:4096 at day 11 and 9, respectively. The bioassays revealed the presence of the parasite in semen samples of a boar inoculated with oocysts (GI) at 3, 49 and 56 DPI and from two boars infected with tachyzoites (GII), one animal at 5 and two animals at 49 days DPI. Mice inoculated with semen from the control group (GIII) remained serologically negative. PCR analysis showed T. gondii DNA in the semen of Boar 1 and Boar 3 inoculated with tachyzoites and oocysts, respectively. The immuno-histochemical tests showed T. gondii in the reproductive organs of Boar 1 and Boar 2, inoculated with tachyzoites and oocysts, respectively. These findings suggest the possible occurrence of venereal transmission of T. gondii in swine.

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.