Autochthonous canine visceral leishmaniasis cases occur in Paraná state since 2012: isolation and identification of Leishmania infantum / Leishmaniose visceral canina autóctone ocorre no estado do Paraná desde 2012: isolamento e identificação de Leishmania infantum

Abstract The aim of this study was to confirm the emergence of canine visceral leishmaniasis among dogs in Foz do Iguaçu. The disease was diagnosed through the isolation and molecular identification of Leishmania infantum. In the first sample collection stage (2012), three lymph node aspirates and 46 buffy coat samples were obtained mostly from the dogs that were seroreagents for leishmaniasis. In the second sample collection stage (2013), the buffy coat samples were collected from 376 dogs located close to Paraguay, Paraná river, center and peripheral parts of the city. The DNA from the six isolates, four from the first sampling stage (4/49) and two from the second sampling stage (2/376), was subjected to polymerase chain reaction using the K26F/R primers. The isolate was confirmed as L. infantum by sequencing. As none of the dogs had ever left the city, the isolates were confirmed as autochthonous. Further, the study confirmed the emergence of canine visceral leishmaniasis in Paraná through the identification of L. infantum among dogs in Foz do Iguaçu city. Hence, collaborative control measures should be designed and implemented by the public agencies and research institutions of Brazil, Argentina, and Paraguay to control the spread of visceral leishmaniasis.

Resumo O objetivo deste estudo foi confirmar a emergência da leishmaniose visceral canina em Foz do Iguaçu próximo à fronteira com a Argentina e ao Paraguai, por meio do isolamento e identificação molecular de Leishmania infantum. Em um primeiro estágio de coleta de animais (2012), três amostras de aspirados de linfonodos e 46 camadas leucocitárias foram obtidas de cães soropositivos para leishmaniose. Em um segundo estágio de coleta (2013), foram coletadas amostras de camada leucocitária de 376 cães de 20 localidades próximas à fronteira com o Paraguai, rio Paraná, centro e periferia da cidade. Seis isolados foram obtidos, quatro da primeira etapa (4/49) e dois da segunda etapa (2/376); estes isolados foram submetidos à amplificação com iniciadores K26F/R, e a análise de sua sequência confirmou a espécie como L. infantum. A autoctonia dos casos foi confirmada, pois 100% dos cães nunca haviam saído da cidade. O estudo confirma a emergência de leishmaniose visceral canina no Paraná com identificação de L. infantum em cães da cidade de Foz do Iguaçu. Assim, medidas de controle devem ser elaboradas e implementadas por órgãos públicos e instituições de pesquisa do Brasil, Argentina e Paraguai em parceria com o objetivo de controlar a disseminação de zoonoses e os casos humanos de LV.

Autochthonous canine visceral leishmaniasis cases occur in Paraná state since 2012: isolation and identification of Leishmania infantum / Leishmaniose visceral canina autóctone ocorre no estado do Paraná desde 2012: isolamento e identificação de Leishmania infantum

The aim of this study was to confirm the emergence of canine visceral leishmaniasis among dogs in Foz do Iguaçu. The disease was diagnosed through the isolation and molecular identification of Leishmania infantum. In the first sample collection stage (2012), three lymph node aspirates and 46 buffy coat samples were obtained mostly from the dogs that were seroreagents for leishmaniasis. In the second sample collection stage (2013), the buffy coat samples were collected from 376 dogs located close to Paraguay, Paraná river, center and peripheral parts of the city. The DNA from the six isolates, four from the first sampling stage (4/49) and two from the second sampling stage (2/376), was subjected to polymerase chain reaction using the K26F/R primers. The isolate was confirmed as L. infantum by sequencing. As none of the dogs had ever left the city, the isolates were confirmed as autochthonous. Further, the study confirmed the emergence of canine visceral leishmaniasis in Paraná through the identification of L. infantum among dogs in Foz do Iguaçu city. Hence, collaborative control measures should be designed and implemented by the public agencies and research institutions of Brazil, Argentina, and Paraguay to control the spread of visceral leishmaniasis.(AU)

O objetivo deste estudo foi confirmar a emergência da leishmaniose visceral canina em Foz do Iguaçu próximo à fronteira com a Argentina e ao Paraguai, por meio do isolamento e identificação molecular de Leishmania infantum. Em um primeiro estágio de coleta de animais (2012), três amostras de aspirados de linfonodos e 46 camadas leucocitárias foram obtidas de cães soropositivos para leishmaniose. Em um segundo estágio de coleta (2013), foram coletadas amostras de camada leucocitária de 376 cães de 20 localidades próximas à fronteira com o Paraguai, rio Paraná, centro e periferia da cidade. Seis isolados foram obtidos, quatro da primeira etapa (4/49) e dois da segunda etapa (2/376); estes isolados foram submetidos à amplificação com iniciadores K26F/R, e a análise de sua sequência confirmou a espécie como L. infantum. A autoctonia dos casos foi confirmada, pois 100% dos cães nunca haviam saído da cidade. O estudo confirma a emergência de leishmaniose visceral canina no Paraná com identificação de L. infantum em cães da cidade de Foz do Iguaçu. Assim, medidas de controle devem ser elaboradas e implementadas por órgãos públicos e instituições de pesquisa do Brasil, Argentina e Paraguai em parceria com o objetivo de controlar a disseminação de zoonoses e os casos humanos de LV.(AU)

Autochthonous canine visceral leishmaniasis cases occur in Paraná state since 2012: isolation and identification of Leishmania infantum.

The aim of this study was to confirm the emergence of canine visceral leishmaniasis among dogs in Foz do Iguaçu. The disease was diagnosed through the isolation and molecular identification of Leishmania infantum. In the first sample collection stage (2012), three lymph node aspirates and 46 buffy coat samples were obtained mostly from the dogs that were seroreagents for leishmaniasis. In the second sample collection stage (2013), the buffy coat samples were collected from 376 dogs located close to Paraguay, Paraná river, center and peripheral parts of the city. The DNA from the six isolates, four from the first sampling stage (4/49) and two from the second sampling stage (2/376), was subjected to polymerase chain reaction using the K26F/R primers. The isolate was confirmed as L. infantum by sequencing. As none of the dogs had ever left the city, the isolates were confirmed as autochthonous. Further, the study confirmed the emergence of canine visceral leishmaniasis in Paraná through the identification of L. infantum among dogs in Foz do Iguaçu city. Hence, collaborative control measures should be designed and implemented by the public agencies and research institutions of Brazil, Argentina, and Paraguay to control the spread of visceral leishmaniasis.

Variables associated with the prevalence of anti-Leishmania spp. antibodies in dogs on the tri-border of Foz do Iguaçu, Paraná, Brazil.

The aim of this study was to investigate the occurrence of anti-Leishmania spp. antibodies in dogs from localities in the city of Foz do Iguaçu, Paraná state, Brazil, on the border with Argentina and Paraguay. Blood samples dogs were collected to perform the following serologic tests: immunochromatographic DPP® rapid test, indirect immunoenzymatic assay (ELISA) and indirect immunofluorescence assay (IFA). In 2012, 285 dogs were analyzed on Argentina border, and in 2013, serum samples from 396 dogs on the border of Paraguay were collected. Using ELISA for screening and IFA for the confirmatory test, the results showed that the antibody prevalence was 1.8% (5/285) on the border of Argentina and 3.0% (12/396) on Paraguay border. When using the DPP® for screening and ELISA as a confirmatory analysis, we observed a seroreagent prevalence in dogs of 2.5% (7/285) on Argentina border and 5.1% (20/396) on Paraguay border. The non-public collection of domestic waste (p= 0.0004) was shown to be associated with leishmaniasis. This study shows the presence of leishmaniasis and suggest the emergence of canine visceral leishmaniasis in state of Paraná due to the confirmed occurrence of seroreactive dogs on Argentina and Paraguay border, which has environmental and geographical characteristics that favor the spread of the parasite.

Variables associated with the prevalence of anti-Leishmania spp. antibodies in dogs on the tri-border of Foz do Iguaçu, Paraná, Brazil / Variáveis associadas à prevalência de anticorpos anti- Leishmania spp. em cães na tríplice fronteira de Foz do Iguaçu, Paraná, Brasil

Abstract The aim of this study was to investigate the occurrence of anti-Leishmania spp. antibodies in dogs from localities in the city of Foz do Iguaçu, Paraná state, Brazil, on the border with Argentina and Paraguay. Blood samples dogs were collected to perform the following serologic tests: immunochromatographic DPP® rapid test, indirect immunoenzymatic assay (ELISA) and indirect immunofluorescence assay (IFA). In 2012, 285 dogs were analyzed on Argentina border, and in 2013, serum samples from 396 dogs on the border of Paraguay were collected. Using ELISA for screening and IFA for the confirmatory test, the results showed that the antibody prevalence was 1.8% (5/285) on the border of Argentina and 3.0% (12/396) on Paraguay border. When using the DPP® for screening and ELISA as a confirmatory analysis, we observed a seroreagent prevalence in dogs of 2.5% (7/285) on Argentina border and 5.1% (20/396) on Paraguay border. The non-public collection of domestic waste (p= 0.0004) was shown to be associated with leishmaniasis. This study shows the presence of leishmaniasis and suggest the emergence of canine visceral leishmaniasis in state of Paraná due to the confirmed occurrence of seroreactive dogs on Argentina and Paraguay border, which has environmental and geographical characteristics that favor the spread of the parasite.

Resumo O objetivo deste estudo foi investigar a ocorrência de anticorpos anti- Leishmania spp. em cães da cidade de Foz do Iguaçu, estado do Paraná, Brasil, fronteira com a Argentina e o Paraguai. Amostras de sangue de cães foram coletadas para realização dos seguintes testes sorológicos: teste rápido imunocromatográfico DPP®, ensaio imunoenzimático indireto (ELISA) e ensaio de imunofluorescência indireta (IFI). Em 2012, 285 cães foram analisados na fronteira com Argentina e, em 2013, amostras de soro de 396 cães na fronteira com o Paraguai. Utilizando ELISA para triagem e IFA para o teste confirmatório, os resultados mostraram uma prevalência de anticorpos de 1,8% (5/285) na fronteira da Argentina e 3,0% (12/396) na fronteira com o Paraguai. Ao usar o DPP® para triagem e ELISA como uma análise confirmatória, observou-se uma prevalência de cães sororreagentes de 2,5% (7/285) na fronteira com a Argentina e 5,1% (20/396) na fronteira com o Paraguai. A não coleta pública de lixo doméstico (p = 0,0004) mostrou-se associada à leishmaniose. Este estudo demonstra a presença de leishmaniose e sugere a emergência da leishmaniose visceral canina no estado do Paraná devido à ocorrência de cães sororreagentes na fronteira Argentina e Paraguai, que possuem características ambientais e geográficas que favorecem a disseminação do parasito.

Variables associated with the prevalence of anti-Leishmania spp. antibodies in dogs on the tri-border of Foz do Iguaçu, Paraná, Brazil / Variáveis associadas à prevalência de anticorpos anti- Leishmania spp. em cães na tríplice fronteira de Foz do Iguaçu, Paraná, Brasil

The aim of this study was to investigate the occurrence of anti-Leishmania spp. antibodies in dogs from localities in the city of Foz do Iguaçu, Paraná state, Brazil, on the border with Argentina and Paraguay. Blood samples dogs were collected to perform the following serologic tests: immunochromatographic DPP® rapid test, indirect immunoenzymatic assay (ELISA) and indirect immunofluorescence assay (IFA). In 2012, 285 dogs were analyzed on Argentina border, and in 2013, serum samples from 396 dogs on the border of Paraguay were collected. Using ELISA for screening and IFA for the confirmatory test, the results showed that the antibody prevalence was 1.8% (5/285) on the border of Argentina and 3.0% (12/396) on Paraguay border. When using the DPP® for screening and ELISA as a confirmatory analysis, we observed a seroreagent prevalence in dogs of 2.5% (7/285) on Argentina border and 5.1% (20/396) on Paraguay border. The non-public collection of domestic waste (p= 0.0004) was shown to be associated with leishmaniasis. This study shows the presence of leishmaniasis and suggest the emergence of canine visceral leishmaniasis in state of Paraná due to the confirmed occurrence of seroreactive dogs on Argentina and Paraguay border, which has environmental and geographical characteristics that favor the spread of the parasite.(AU)

O objetivo deste estudo foi investigar a ocorrência de anticorpos anti- Leishmania spp. em cães da cidade de Foz do Iguaçu, estado do Paraná, Brasil, fronteira com a Argentina e o Paraguai. Amostras de sangue de cães foram coletadas para realização dos seguintes testes sorológicos: teste rápido imunocromatográfico DPP®, ensaio imunoenzimático indireto (ELISA) e ensaio de imunofluorescência indireta (IFI). Em 2012, 285 cães foram analisados na fronteira com Argentina e, em 2013, amostras de soro de 396 cães na fronteira com o Paraguai. Utilizando ELISA para triagem e IFA para o teste confirmatório, os resultados mostraram uma prevalência de anticorpos de 1,8% (5/285) na fronteira da Argentina e 3,0% (12/396) na fronteira com o Paraguai. Ao usar o DPP® para triagem e ELISA como uma análise confirmatória, observou-se uma prevalência de cães sororreagentes de 2,5% (7/285) na fronteira com a Argentina e 5,1% (20/396) na fronteira com o Paraguai. A não coleta pública de lixo doméstico (p = 0,0004) mostrou-se associada à leishmaniose. Este estudo demonstra a presença de leishmaniose e sugere a emergência da leishmaniose visceral canina no estado do Paraná devido à ocorrência de cães sororreagentes na fronteira Argentina e Paraguai, que possuem características ambientais e geográficas que favorecem a disseminação do parasito.(AU)

Leishmaniasis in dogs from recycling centers and from a neighborhood with adjacent forest in an urban area of Londrina, Paraná, Brazil / Leishmaniose em cães de centros de reciclagem e de um bairro nas adjacências de uma mata urbana em Londrina, PR, Brasil

The leishmaniasis are a group of globally widespread parasitic diseases caused by a protozoa of the genus Leishmania, the state of Paraná accounts for 98% of leishmaniasis human cases in southern Brazil and the dogs might serve as a sentinel of the risk of infection to humans in a given area. The aim of this work was to study the epidemiology of leishmaniasis in dogs residents in two different environments, they are: recycling of solid waste centers (Group 1) and a neighborhood adjacent to a forest (Group 2), both in Londrina, Paraná State, Brazil and to determine epidemiological variables associated to infection in these different realities. Serum samples, randomly collected, from 61 dogs from Group 1 and from 74 dogs from Group 2 were submitted to Indirect Immunofluorescence Antibody Test (IFAT) to research for anti-Leishmania IgG antibodies. Of the 135 dogs analyzed, 13 animals (9.6%) were positive; nine (14.7%) in Group 1 and four (5.4%) in Group 2, the difference between the groups is not statistically significant. These results suggest the presence of the protozoan Leishmania spp. in both environments studied which indicate potential for infection in the human population.(AU)

As leishmanioses são um grupo de doenças parasitárias disseminadas por todo o mundo, causadas por protozoários do gênero Leishmania, o estado do Paraná é responsável por 98% dos casos de leishmaniose em humanos do Sul do Brasil e os cães podem servir como sentinelas para o risco de infecção em humanos em uma dada área. O objetivo desse trabalho foi estudar a epidemiologia da leishmaniose em cães residentes em dois diferentes ambientes, são eles: centros de reciclagem de resíduos sólidos (Grupo 1) e um bairro adjacente a uma floresta (Grupo 2) em Londrina/PR, Brasil e determiner variáveis epidemiológicas asociadas à infecção nessas diferentes realidades. Amostras de soro, randomicamente coletadas, de 61 cães do Grupo 1 e de 74 cães do Grupo 2 foram submetidos à Reação de Imunofluorescência Indireta (RIFI) para pesquisa de anticorpos IgG anti-Leishmania. Dos 135 cães analisados, 13 (9,6%) foram positivos; 9 (14,7%) no Grupo 1 e 4 (5,4%) no Grupo 2, a diferença entre os grupos não foi estatisticamente significante. Esses resultados sugerem a presença do protozoário Leishmania spp. nos dois ambientes estudados o que indica potencial de infecção na população humana.(AU)

Leishmaniasis in dogs from recycling centers and from a neighborhood with adjacent forest in an urban area of Londrina, Paraná, Brazil / Leishmaniose em cães de centros de reciclagem e de um bairro nas adjacências de uma mata urbana em Londrina, PR, Brasil

The leishmaniasis are a group of globally widespread parasitic diseases caused by a protozoa of the genus Leishmania, the state of Paraná accounts for 98% of leishmaniasis human cases in southern Brazil and the dogs might serve as a sentinel of the risk of infection to humans in a given area. The aim of this work was to study the epidemiology of leishmaniasis in dogs residents in two different environments, they are: recycling of solid waste centers (Group 1) and a neighborhood adjacent to a forest (Group 2), both in Londrina, Paraná State, Brazil and to determine epidemiological variables associated to infection in these different realities. Serum samples, randomly collected, from 61 dogs from Group 1 and from 74 dogs from Group 2 were submitted to Indirect Immunofluorescence Antibody Test (IFAT) to research for anti-Leishmania IgG antibodies. Of the 135 dogs analyzed, 13 animals (9.6%) were positive; nine (14.7%) in Group 1 and four (5.4%) in Group 2, the difference between the groups is not statistically significant. These results suggest the presence of the protozoan Leishmania spp. in both environments studied which indicate potential for infection in the human population.

As leishmanioses são um grupo de doenças parasitárias disseminadas por todo o mundo, causadas por protozoários do gênero Leishmania, o estado do Paraná é responsável por 98% dos casos de leishmaniose em humanos do Sul do Brasil e os cães podem servir como sentinelas para o risco de infecção em humanos em uma dada área. O objetivo desse trabalho foi estudar a epidemiologia da leishmaniose em cães residentes em dois diferentes ambientes, são eles: centros de reciclagem de resíduos sólidos (Grupo 1) e um bairro adjacente a uma floresta (Grupo 2) em Londrina/PR, Brasil e determiner variáveis epidemiológicas asociadas à infecção nessas diferentes realidades. Amostras de soro, randomicamente coletadas, de 61 cães do Grupo 1 e de 74 cães do Grupo 2 foram submetidos à Reação de Imunofluorescência Indireta (RIFI) para pesquisa de anticorpos IgG anti-Leishmania. Dos 135 cães analisados, 13 (9,6%) foram positivos; 9 (14,7%) no Grupo 1 e 4 (5,4%) no Grupo 2, a diferença entre os grupos não foi estatisticamente significante. Esses resultados sugerem a presença do protozoário Leishmania spp. nos dois ambientes estudados o que indica potencial de infecção na população humana.

Zoonoses em cães e gatos atendidos em Projeto de Controle de Natalidade: Toxoplasma gondii, Leishmania spp. e Leptospira spp., sorodiagnóstico e epidemiologia / Zoonosis in dogs and cats attended by the Birth Control Project: Toxoplasma gondii, Leishmania spp. and Leptospira spp., serodiagnosis and epidemiology

A proximidade da relação homem-animal tem aumentado o risco de transmissão de doenças de caráter zoonótico. O objetivo desse trabalho foi determinar a prevalência de anticorpos IgG anti-Toxoplasma gondii, anti- Leishmania spp. e anti- Leptospira spp. nos cães e gatos atendidos pelo Projeto Controle de Natalidade (PCN) nos anos de 2004 a 2012 e verificar a significância estatística de variáveis ambientais e da população de estudo à presença dos agentes infecciosos. Amostras de sangue de 271 cães e 415 gatos foram colhidas por venopunção jugular ou cefálica para a posterior obtenção dos soros. Estes foram submetidos à técnica de imunofluorescência indireta para detecção de anticorpos IgG anti-T. gondii e anti-Leishmania spp. e à técnica de soroaglutinação microscópica para detecção de anticorpos anti- Leptospira spp. A tabulação dos dados foi realizada utilizando-se o pacote estatístico EpiInfo (3.4.3). Dos soros caninos examinados, 20,6% (56/271), 1,3% (3/236) e 21,6% (51/236) foram positivos para T. gondii, Leishmania spp. e Leptospira spp., respectivamente. Encontrou-se maior prevalência de anticorpos anti- T. gondii em cães sem raça definida (p=0, 0009), em cães alimentados com carne crua ou mal cozida (p= 0,0101) e naqueles que não receberam vacina polivalente (p= 0,0147). Não houve associação estatística entre as variáveis avaliadas no questionário epidemiológico para leishmaniose e leptospirose. Dos soros felinos analisados, 20,9% (87/415) foram positivos para T. gondii. Maiores prevalências foram observadas nos felinos com hábito de caçar roedores (p= 0,0388) e que não recebem água tratada (p= 0,0294). Os resultados obtidos confirmam a importância dos fatores de risco ingestão de carne crua ou mal cozida, ingestão de água não tratada e predação de roedores para infecção por T. gondii. Demonstrou-se baixa prevalência ao sorodiagnostico para Leishmania spp. Os sorovares de Leptospira spp. mais frequentes foram Copenhageni, Canicola e Butembo.(AU)

Increasing human-animal proximity has also increased risks of zoonotic disease transmission. This study measured the prevalence of anti-Toxoplasma gondii, anti-Leishmania spp. and anti-Leptospira spp. IgG antibodies in dogs and cats attended by the Birth Control Project (BCP) from 2004 to 2012 to determine which environmental variables were significantly associated with infectious agents in the study population. Blood samples from 271 dogs and 415 cats were collected by jugular or cephalic venipuncture for serological analysis. Indirect immunofluorescence assays were used to detect anti-T. gondii and anti-Leishmania spp. IgG antibodies and microscopic serum-agglutination tests were used to detect anti- Leptospira spp. antibodies. Data tabulation and statistical analyses were performed using EpiInfo (3.4.3). Among canine sera samples, 20.6% (56/271), 1.3% (3/236), and 21.6% (51/236) were positive for T. gondii, Leishmania spp., and Leptospira spp., respectively. Higher T. gondii prevalence was found in dogs of unknown breeding history (p=0.0009), those fed raw or undercooked meat (p=0.0101), and those that did not receive the polyvalent vaccine (p=0.0147). There was no statistically significant association between leishmaniosis and leptospirosis and variables evaluated in the epidemiological questionnaire. Among feline sera samples, 20.9% (87/415) were positive for T. gondii. The highest prevalence rates were observed in felines that hunted rodents (p=0.0388) and that consumed untreated water (p=0.0294). These results confirmed that ingestion of raw or undercooked meat, drinking untreated water, and predation of rodents were important risk factors for T. gondii infection. A low Leishmania spp. serodiagnostic prevalence was observed. The most frequent Leptospira spp. serovars were Copenhageni, Canicola, and Butembo.(AU)

Zoonoses em cães e gatos atendidos em Projeto de Controle de Natalidade: Toxoplasma gondii, Leishmania spp. e Leptospira spp., sorodiagnóstico e epidemiologia / Zoonosis in dogs and cats attended by the Birth Control Project: Toxoplasma gondii, Leishmania spp. and Leptospira spp., serodiagnosis and epidemiology

A proximidade da relação homem-animal tem aumentado o risco de transmissão de doenças de caráter zoonótico. O objetivo desse trabalho foi determinar a prevalência de anticorpos IgG anti-Toxoplasma gondii, anti- Leishmania spp. e anti- Leptospira spp. nos cães e gatos atendidos pelo Projeto Controle de Natalidade (PCN) nos anos de 2004 a 2012 e verificar a significância estatística de variáveis ambientais e da população de estudo à presença dos agentes infecciosos. Amostras de sangue de 271 cães e 415 gatos foram colhidas por venopunção jugular ou cefálica para a posterior obtenção dos soros. Estes foram submetidos à técnica de imunofluorescência indireta para detecção de anticorpos IgG anti-T. gondii e anti-Leishmania spp. e à técnica de soroaglutinação microscópica para detecção de anticorpos anti- Leptospira spp. A tabulação dos dados foi realizada utilizando-se o pacote estatístico EpiInfo (3.4.3). Dos soros caninos examinados, 20,6% (56/271), 1,3% (3/236) e 21,6% (51/236) foram positivos para T. gondii, Leishmania spp. e Leptospira spp., respectivamente. Encontrou-se maior prevalência de anticorpos anti- T. gondii em cães sem raça definida (p=0, 0009), em cães alimentados com carne crua ou mal cozida (p= 0,0101) e naqueles que não receberam vacina polivalente (p= 0,0147). Não houve associação estatística entre as variáveis avaliadas no questionário epidemiológico para leishmaniose e leptospirose. Dos soros felinos analisados, 20,9% (87/415) foram positivos para T. gondii. Maiores prevalências foram observadas nos felinos com hábito de caçar roedores (p= 0,0388) e que não recebem água tratada (p= 0,0294). Os resultados obtidos confirmam a importância dos fatores de risco ingestão de carne crua ou mal cozida, ingestão de água não tratada e predação de roedores para infecção por T. gondii. Demonstrou-se baixa prevalência ao sorodiagnostico para Leishmania spp. Os sorovares de Leptospira spp. mais frequentes foram Copenhageni, Canicola e Butembo.

Increasing human-animal proximity has also increased risks of zoonotic disease transmission. This study measured the prevalence of anti-Toxoplasma gondii, anti-Leishmania spp. and anti-Leptospira spp. IgG antibodies in dogs and cats attended by the Birth Control Project (BCP) from 2004 to 2012 to determine which environmental variables were significantly associated with infectious agents in the study population. Blood samples from 271 dogs and 415 cats were collected by jugular or cephalic venipuncture for serological analysis. Indirect immunofluorescence assays were used to detect anti-T. gondii and anti-Leishmania spp. IgG antibodies and microscopic serum-agglutination tests were used to detect anti- Leptospira spp. antibodies. Data tabulation and statistical analyses were performed using EpiInfo (3.4.3). Among canine sera samples, 20.6% (56/271), 1.3% (3/236), and 21.6% (51/236) were positive for T. gondii, Leishmania spp., and Leptospira spp., respectively. Higher T. gondii prevalence was found in dogs of unknown breeding history (p=0.0009), those fed raw or undercooked meat (p=0.0101), and those that did not receive the polyvalent vaccine (p=0.0147). There was no statistically significant association between leishmaniosis and leptospirosis and variables evaluated in the epidemiological questionnaire. Among feline sera samples, 20.9% (87/415) were positive for T. gondii. The highest prevalence rates were observed in felines that hunted rodents (p=0.0388) and that consumed untreated water (p=0.0294). These results confirmed that ingestion of raw or undercooked meat, drinking untreated water, and predation of rodents were important risk factors for T. gondii infection. A low Leishmania spp. serodiagnostic prevalence was observed. The most frequent Leptospira spp. serovars were Copenhageni, Canicola, and Butembo.

Gestational toxoplasmosis in Paraná State, Brazil: prevalence of IgG antibodies and associated risk factors

The aim of the present study was to verify the association between seropositivity for IgG anti-Toxoplasma gondii antibodies and social, economic and environmental variables of pregnant women attending the public health centers of Paraná, Brazil. From January 2007 to July 2010, 2226 pregnant women were interviewed and detection of anti-T. gondii specific IgG and IgM antibodies was performed by chemiluminescence test. Seropositivity for anti-T. gondii IgG was observed in 1151 (51.7%) pregnant women, 29 of which (1.3%) presented IgM reagent with IgG of high avidity. The variables associated with the presence of IgG were residency in the rural area, more than one pregnancy, less than or equal to eight years schooling, low per capita income, age group, raw or poorly cooked meat ingestion, and contact with the soil. There was neither association with raw fruit and vegetable ingestion nor with the presence of cats in the residencies.

Gestational toxoplasmosis in Paraná State, Brazil: prevalence of IgG antibodies and associated risk factors.

The aim of the present study was to verify the association between seropositivity for IgG anti-Toxoplasma gondii antibodies and social, economic and environmental variables of pregnant women attending the public health centers of Paraná, Brazil. From January 2007 to July 2010, 2226 pregnant women were interviewed and detection of anti-T. gondii specific IgG and IgM antibodies was performed by chemiluminescence test. Seropositivity for anti-T. gondii IgG was observed in 1151 (51.7%) pregnant women, 29 of which (1.3%) presented IgM reagent with IgG of high avidity. The variables associated with the presence of IgG were residency in the rural area, more than one pregnancy, less than or equal to eight years schooling, low per capita income, age group, raw or poorly cooked meat ingestion, and contact with the soil. There was neither association with raw fruit and vegetable ingestion nor with the presence of cats in the residencies.

Factors associated to infection by Toxoplasma gondii in pregnant women attended in Basic Health Units in the city of Rolândia, Paraná, Brazil.

The aim of the present work was to determine the prevalence of IgG and IgM anti-Toxoplasma gondii antibodies and the factors associated to the infection in pregnant women attended in Basic Health Units in Rolândia, Paraná, Brazil. The sample was divided in two groups: group I (320 pregnant women who were analyzed from July 2007 to February 2008) and group II (287 pregnant women who were analyzed from March to October 2008). In group I, it was found 53.1% of pregnant women with IgG reactive and IgM non-reactive, 1.9% with IgG and IgM reactive, 0.3% with IgG non-reactive and IgM reactive and 44.7% with IgG and IgM non-reactive. In group II, it was found 55.1% with IgG reactive and IgM non-reactive and 44.9% with IgG and IgM non-reactive. The variables associated to the presence of IgG antibodies were: residence in rural areas, pregnant women between 35-40 years old, low educational level, low family income, more than one pregnancy, drinking water which does not originate from the public water supply system and the habit of handling soil or sand. Guidance on primary prevention measures and the quarterly serological monitoring of the pregnant women in the risk group are important measures to prevent congenital toxoplasmosis.

Factors associated to infection by Toxoplasma gondii in pregnant women attended in Basic Health Units in the city of Rolândia, Paraná, Brazil / Fatores associados à infecção por Toxoplasma gondii em gestantes atendidas nas Unidades Básicas de Saúde do município de Rolândia, Paraná, Brasil

The aim of the present work was to determine the prevalence of IgG and IgM anti-Toxoplasma gondii antibodies and the factors associated to the infection in pregnant women attended in Basic Health Units in Rolândia, Paraná, Brazil. The sample was divided in two groups: group I (320 pregnant women who were analyzed from July 2007 to February 2008) and group II (287 pregnant women who were analyzed from March to October 2008). In group I, it was found 53.1 percent of pregnant women with IgG reactive and IgM non-reactive, 1.9 percent with IgG and IgM reactive, 0.3 percent with IgG non-reactive and IgM reactive and 44.7 percent with IgG and IgM non-reactive. In group II, it was found 55.1 percent with IgG reactive and IgM non-reactive and 44.9 percent with IgG and IgM non-reactive. The variables associated to the presence of IgG antibodies were: residence in rural areas, pregnant women between 35-40 years old, low educational level, low family income, more than one pregnancy, drinking water which does not originate from the public water supply system and the habit of handling soil or sand. Guidance on primary prevention measures and the quarterly serological monitoring of the pregnant women in the risk group are important measures to prevent congenital toxoplasmosis.

O objetivo deste trabalho foi determinar a prevalência de anticorpos IgG e IgM anti-Toxoplasma gondii e os fatores associados à infecção em gestantes atendidas nas Unidades Básicas de Saúde de Rolândia, Paraná, Brasil. A amostra foi dividida em dois grupos: grupo I (320 gestantes, analisadas entre julho de 2007 e fevereiro de 2008) e grupo II (287 gestantes, analisadas de março a outubro de 2008). No grupo I, foram encontrados 53,1 por cento de gestantes IgG reagente e IgM não reagente, 1,9 por cento de IgG e IgM reagentes, 0,3 por cento de IgG não reagente e IgM reagente e 44,7 por cento de IgG e IgM não reagentes. No grupo II foram 55,1 por cento de IgG reagente e IgM não reagente e 44,9 por cento de IgG e IgM não reagentes. As variáveis associadas à infecção foram: residência em área rural, gestantes entre 35-40 anos de idade, baixa escolaridade, baixa renda familiar, mais de uma gravidez, consumo de água não proveniente do sistema de abastecimento público de água e hábito de manipular terra ou areia. As orientações sobre medidas de prevenção primária e o acompanhamento sorológico trimestral das gestantes no grupo de risco são medidas importantes para prevenir a toxoplasmose congênita.

Protective activity against oocyst shedding in cats vaccinated with crude rhoptry proteins of the Toxoplasma gondii by the intranasal route.

This study evaluated a vaccine made from crude rhoptry proteins of Toxoplasma gondii with Quil-A, which was administered to cats by the intranasal route. Eleven short-hair domestic cats were divided into four groups: G1 (n=3) received three doses (200 microg/dose) of the rhoptry vaccine with Quil-A (20 microg); G2 (n=3) received PBS with Quil-A (20 microg); G3 (n=3) and G4 (n=2) received only PBS. Treatments were administered at days 0, 21, and 42 by the intranasal route. Challenge was done to G1, G2, and G3 animals with 600 cysts of the VEG strain on day 51 (challenge day); G4 animals were unchallenged. The anti-T. gondii IgG and IgA antibody levels from sera were measured by indirect enzyme-linked immunosorbent assay (ELISA). At challenge, two animals from G1 revealed antibody levels for both IgG and IgA; oocysts were not detected in feces of these two cats. There were no differences in hematological values between groups throughout the experiment (p>0.10). Preventable fractions were 67% in G1 and 0% in G2 and G3. Comparatively, G1 animals shed 89.3% and 90.8% less oocysts than G3 and G4, respectively. Two out of three cats were protected against T. gondii oocyst shedding when the rhoptry vaccine was administered by the intranasal route. This is the first study using crude rhoptry proteins as vaccine by the intranasal route in cats to evaluate protection against oocysts shedding.