Leptospira spp. in horses in southern Brazil: Seroprevalence, infection risk factors, and influence on reproduction.

Leptospirosis in horses is often associated with reproductive disorders. In the southern states of Brazil, horses are used for various jobs and cultural practices; nevertheless, serological surveillance for Leptospira is rare. Therefore, the objective of this study was to determine the seroprevalence of Leptospira spp. in horses in southern Brazil, as well as to identify the risk factors for infection and its impacts on reproduction. We performed microscopic agglutination tests for 12 serovars that corresponding 9 serogroup (Sejroe, Icterohaemorrhagiae, Australis, Pyrogenes, Pomona, Canicola, Grippotyphosa, Tarassovi and Ballum) in 595 samples from 60 herds. A brief history was obtained to analyze risk factors for reproductive disorders. A total of 45.9% of the tested horses were seropositive, of which the most frequent serogroups were Icterohaemorrhagiae (Icterohaemorrhagiae and Copenhageni serovars) and Ballum (Ballum serovar). Simple infections were found in 45.4% of seropositive animals, while mixed infections occurred in 54.6% of horses. There was a correlation between seropositivity and age and sex, that is, seropositivity was more frequent in animals over 6 years old and in females. There was no correlation between seropositivity and reproductive disorders. We conclude that there is a high seroprevalence of Leptospira spp. in southern Brazil with predominance of Icterohaemorrhagiae serogroup, mainly in older animals. Location, breeds, contact with dogs or other domestic animals are not risk factors, whereas gender is a risk factor. Reproductive disorders are not due to leptospirosis in the study region.

Relation of reproductive disturbance in sheep and Leptospira interrogans serovar Icterohaemorrhagiae infection: Impacts on cellular oxidation status.

This study evaluated the seropositivity of Brucella abortus and Leptospira interrogans in ewes with reproductive disturbances in southern Brazil and verified the creatine kinase (CK) activity and oxidation status via assessment of superoxide dismutase, glutathione peroxidase and glutathione transferase in serum of seropositive animals for L. interrogans serovar Icterohaemorrhagiae. For Leptospira infection 381 animals with clinical history of reproductive disturbance from Planalto Serrano de Santa Catarina (Brazil) were analyzed, showing an occurrence for L. interrogans of 20.2% from which 81.8% were seropositive for L. interrogans Icterohaemorrhagiae. Serovars Wolfii, Grippothyphosa, Bratislava, Canicola and Butembo were also identified. In the case of B. abortus, positive cases were identified by buffered acidified antigen, finding 14 positive samples, but none of them were positive after a second test (2-mercaptoethanol), showing the absence of relationship between infection with B. abortus and abortion in the tested individuals. Serum reactive oxygen species (ROS) levels and CK activity were found higher in animals positive for Leptospira infection, presenting higher titrations (1:320) than non-infected individuals. Serum glutathione peroxidase activity was higher in positive animals with titrations 1:160 and 1:320, while serum glutathione S-transferase was higher in positive individuals only for titrations 1:320. Serum superoxide dismutase showed lower activity in infected animals with titrations of 1:320. Our results show the region of Planalto Serrano de Santa Catarina with a high occurrence levels of sheep infected by L. interrogans serovar Icterohaemorrhagiae, from which animals with high titrations (1:320) present oxidative stress elicited by excessive ROS production, triggering the stimulation of antioxidant systems to counter this excess. In summary, ovine with higher titrations (1:320) present oxidative damage that can contribute to disease pathophysiology.

Antibodies to Leptospira interrogans in goats and risk factors of the disease in Santa Catarina (West side), Brazil.

Leptospirosis is an infectious disease caused by the bacterium Leptospira spp. In goats, the productive impact of leptospirosis is not well known and totally unknown in Santa Catarina (SC), Brazil. This study aimed to investigate leptospirosis seroprevalence and its risk factors in goats in the west side of SC. A total of 654 blood samples were analyzed using the microscopic agglutination technique and 35.47% (232) of the animals were seropositives. Except for serogroup Autumnalis, positive samples for all other serogroups were found as follows: Sejroe (Hardjo, Wolffi), Grippotyphosa (Grippotyphosa), Canicola (Canicola), Icterohaemorrhagiae (Icterohaemorrhagiae, Copenhageni), Australis (Australis, Bratislava) and Pomona (Pomona). The contact among sheep and goats, and the addition of concentrate as food supplement were found to be risk factors for leptospirosis. Based on these results, we conclude that there is a high occurrence of anti-Leptospira antibodies in goats in the Western part of Santa Catarina State.

Immunological response and markers of cell damage in seropositive horses for Toxoplasma gondii.

Toxoplasmosis is an important parasitic disease affecting several species of mammals, but little is known about this disease in horses. This study aimed to investigate the levels of several immunological variables and markers of cell damage in the serum of seropositive horses for Toxoplasma gondii. Sera samples of adult horses from the Santa Catarina State, Brazil used on a previous study were divided into groups according to their antibody levels for T. gondii determined by immunofluorescence assay, i.e. 20 samples from seronegative horses (Group A - control), 20 samples from horses with titers of 1:64 (Group B), 20 samples of horses with titers of 1:256 (Group C), and five samples from horses with titers of 1:1024 (Group D). Positive animals (Groups B, C, and D) had higher levels of immunoglobulins (IgM and IgG), pro-inflammatory cytokines (TNF-α, IFN-γ, IL-1, IL-4, and IL-6) and protein C-reactive protein, as well as lower levels of IL-10 (anti-inflammatory cytokine) when compared to seronegative horses (Group A). The nitric oxide levels were also elevated in seropositive horses. Therefore, we have found humoral and cellular immune responses in seropositive horses, and a correlation between high antibody levels and inflammatory mediators. Markers of cell injury by lipid peroxidation (TBARS) and protein oxidation (AOPP) were elevated in animals seropositives for T. gondii when compared to seronegatives. Therefore, seropositive horses to T. gondii can keep active immune responses against the parasite. As a consequence with chronicity of disease, they show cellular lesions that may lead to tissue damage with the appearance of clinical disease.

Toxoplasma gondii antibodies on domiciled cats from Lages municipality, Santa Catarina State, Brazil / Anticorpos contra Toxoplasma gondii em gatos domiciliados no município de Lages, Santa Catarina, Brasil

Sera were collected from 300 domiciled cats from the municipality of Lages, Southern Brazil, to determine the prevalence of Toxoplasma gondii antibodies and risk factors associated. Tests for T. gondii antibodies were performed using indirect immunofluorescent antibody test (IFAT). Positive reactions with titers ≥1:64 were found in 43 (14.33%) cats. A significant number of seropositive cats were ≥6 month old (p = 0.03758) and had access to the streets or/and rural areas (p = 0.04185). The results indicate that T. gondii is widespread in cats in Lages with a prevalence of 14.33%.

Amostras de sangue foram obtidas de 300 gatos domiciliados para determinar a prevalência de anticorpos contra Toxoplasma gondii e possíveis fatores de risco. A detecção de anticorpos da classe IgG foi realizada por meio da Reação de Imunofluorescência Indireta (RIFI). Reações positivas com títulos ≥1:64 foram encontradas em 43 (14,33%) gatos. Os títulos encontrados variaram de 1:64 a 1:4096, sendo que 19 animais (44,19%) tiveram titulação de 1:64, 18 animais (41,86%) com 1:256, cinco animais (11,63%) com 1:1024 e um animal (2,32%) teve titulação de 1:4096. Foi verificada correlação entre soropositividade dos felinos ao T. gondii e idade (p = 0,03758) e com o acesso à rua e/ou zona rural (p = 0,04185). Os resultados indicam que T. gondii está disseminado entre os gatos domiciliados de Lages, Estado de Santa Catarina, Brasil, com uma prevalência de 14,33%.

[Occurrence of antibodies and risk factors for infection for Toxoplasma gondii in dogs in the cities of Lages and Balneário Camboriú, Santa Catarina State, Brazil]. / Ocorrência de anticorpos e fatores de risco para infecção por Toxoplasma gondii em cães, nas cidades de Lages e Balneário Camboriú, Santa Catarina, Brasil.

The aim of this study was to identify the occurrence of antibodies and risk factors for infection for Toxoplasma gondii in dogs in the cities of Lages and Balneário Camboriú, Santa Catarina State, Brazil. Blood samples from 400 domiciliated dogs were processed for antibody detection against T. gondii by indirect immunofuorescence antibody test (IFAT). The following variables were obtained by questionnaire: breed, age, sex, diet, environment, presence of cats and street access. A Chi-Square test was performed to verify the association between the seroprevalence with the studied variables. From all evaluated dogs, 89 (22.3%) had antibodies against T. gondii. A total of 52 (26%) and 37 (18.5%) dogs were positive for T. gondii in Lages and Balneário Camboriú, respectively. No statistical differences were observed on the prevalences of T. gondii between both cities. Greater occurrences of seropositives for toxoplasmosis were observed in mixed-breed dogs (P = 0.002), who had access to the street (P = 0.003) and received homemade diet (P = 0.028). A strong trend for seropositivity against T. gondii was observed among adult dogs, indicating post-natal exposure, and among those that have previous contact with cats.

Ocorrência de anticorpos e fatores de risco para infecção por Toxoplasma gondii em cães, nas cidades de Lages e Balneário Camboriú, Santa Catarina, Brasil / Occurrence of antibodies and risk factors for infection for Toxoplasma gondii in dogs in the cities of Lages and Balneário Camboriú, Santa Catarina State, Brazil

Com os objetivos de conhecer a ocorrência de anticorpos e identificar fatores de risco para a infecção por Toxoplasmagondii em cães, nas cidades de Lages e Balneário Camboriú, SC, amostras de sangue de 400 cães domiciliados foram processadas para a detecção de anticorpos contra T. gondii pela técnica da imunofluorescência indireta (RIFI). Dados referentes à raça, idade, sexo, tipo de dieta, ambiente, presença de gatos e acesso à rua foram obtidos por meio de questionário. Análise estatística foi realizada (teste de χ²). Dos 400 cães, 89 (22,3 por cento) apresentaram anticorpos contra T. gondii. Dos cães de Lages, 52 (26 por cento) foram positivos para T. gondii, enquanto os de Balneário Camboriú, 37 (18,5 por cento)foram sororreagentes. Não foram observadas diferenças estatísticas entre as prevalências de T. gondii nos dois municípios. Maior ocorrência de sororreagentes para toxoplasmose foi verificada entre os cães sem raça definida (P = 0,002), que tinham acesso à rua (P = 0,003) e que recebiam dieta caseira (P = 0,028). Forte tendência para reagentes ao T. gondii foi observada entre os animais adultos, indicando exposição pós-natal ao agente, e entre os cães em contato com felinos.

The aim of this study was to identify the occurrence of antibodies and risk factors for infection for Toxoplasma gondii in dogs in the cities of Lages and Balneário Camboriú, Santa Catarina State, Brazil. Blood samples from 400 domiciliated dogs were processed for antibody detection against T. gondii by indirect immunofluorescence antibody test (IFAT). The following variables were obtained by questionnaire: breed, age, sex, diet, environment, presence of cats and street access. A Chi-Square test was performed to verify the association between the seroprevalence with the studied variables. From all evaluated dogs, 89 (22.3 percent) had antibodies against T. gondii. A total of 52 (26 percent) and 37 (18.5 percent) dogs were positive for T. gondii in Lages and Balneário Camboriú, respectively. No statistical differences were observed on the prevalences of T. gondii between both cities. Greater occurrences of seropositives for toxoplasmosis were observed in mixed-breed dogs (P = 0.002), who had access to the street (P = 0.003) and received homemade diet (P = 0.028). A strong trend for seropositivity against T. gondii was observed among adult dogs, indicating post-natal exposure, and among those that have previous contact with cats.

Toxoplasma gondii in semen of experimentally infected swine

Eight reproductive boars were divided into three groups and inoculated with Toxoplasma gondii [GI (n=3) 1.5x10(4) oocysts strain P; GII (n=3) 1.0x10(6) 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.

Oito reprodutores suínos foram divididos em três grupos e inoculados com Toxoplasma gondii [GI (n=3) 1.5x10(4) oocistos cepa P; GII (n=3) 1.0x10(6) taquizoítos cepa RH, e GIII (n=2) controle, não inoculados]. Exames clínicos, hematológicos, de parasitemia e sorológicos foram realizados para avaliar a infecção toxoplásmica. Pesquisa do parasito no sêmen, por meio do bioensaio e pela técnica da PCR, e em órgãos do sistema reprodutor (bioensaio e imunohistoquímica) foi realizada. Sangue e sêmen foram colhidos nos dias -2, -1, 1, 3, 5, 7, 9, 11, 14, e semanalmente até o 84º dia pós-infecção (DPI). Nenhuma alteração clínica ou hematimétrica foi observadda nos animais. Parasitemia foi detectada em um animal inoculado com oocistos no 7º DPI e em outro inoculado com taquizoítos (GII) nos 3º e 49º DPI. A sorologia revelou a presença de anticorpos contra T. gondii nos animais inoculados com oocistos ou taquizoítos no 7º DPI com títulos de 1:256 e 1:64, que atingiram picos de 1:4096 nos dias 11 e 9, respectivamente. O bioensaio revelou a presença do parasita em amostras seminais de um animal inoculado com oocistos (GI) nos 3º, 49º e 56º DPI, e de dois animais infectados com taquizoítos (GII), um deles no 5º DPI e os dois ao 49º DPI. Pela PCR, o DNA de T. gondii foi detectado no sêmen dos Suínos 1 e 3 inoculados com taquizoítos e oocistos, respectivamente. A imunohistoquímica revelou T. gondii em órgão do aparelho reprodutor dos Suínos 1 e 2, inoculados com taquizoítos e oocistos, respectivamente. Esses achados sugerem a possibilidade da ocorrência da transmissão venérea do T. gondii em suínos.

[Occurrence of anti-Toxoplasma gondii antibodies in swine and ovine slaughtered at municipality of Guarapuava in the State of Paraná, Brazil]. / Ocorrência de anticorpos contra Toxoplasma gondii em suínos e ovinos abatidos no município de Guarapuava, PR, Brasil.

This study evaluated sero-occurrence of toxoplasmosis in swine and ovine slaughtered in the municipality of Guarapuava in the state of Paraná. Blood samples from 117 swine and 157 ovine were collected. Indirect Immunofluorescence Assay (IFA) was used to detect IgG anti-Toxoplasma gondii antibodies. Serum samples were considered positive at dilutions >or=1:64. Out of 117 swine blood sera, 10 (8.54%) were positive, while 11 out of 157 ovine samples (7.0%) showed a reaction to T. gondii antibodies. Our data showed a decrease in the occurrence of toxoplasmosis in swine and ovine. This is due to improved technologies used to raise swine, and better management used to raise ovine as well as the age of the animals evaluated.