Toxoplasma gondii em lingüiças frescas de porco e soroprevalência em açougueiros de fábricas de Londrina, Estado do Paraná, Brasil

Os objetivos deste estudo foram verificar a presença de cistos de Toxoplasma gondii em lingüiça de origem suína tipo frescal e de anticorpos anti-T. gondii no soro de trabalhadores de indústrias produtoras, com Serviço de Inspeção Municipal (SIM) de Londrina, PR. Buscou-se ainda, obter informações relativas às práticas adotadas nestas indústrias e aos hábitos dos trabalhadores, que pudessem influenciar na prevalência desta parasitose. Foram coletadas 149 amostras de lingüiça em oito indústrias produtoras e 47 amostras de sangue de todos os trabalhadores destes locais. Após a realização do bioensaio em camundongos, obteve-se 13 (8,72%) amostras de lingüiça positivas, sendo que em uma o parasita foi isolado e nas outras 12 os camundongos soroconverteram. Dos 47 trabalhadores, 36 (76,6%) atuavam na produção de lingüiça e 11 (23,4%) exerciam outras funções; os percentuais de soropositivos ao T. gondii foram, respectivamente, 59,5% (28/47), 55,5% (20/36) e 72,7% (8/11). Não houve diferença significativa entre nenhuma das variáveis relacionadas às indústrias e aos trabalhadores. Os resultados permitem inferir que lingüiças tipo frescal possuem importância na cadeia epidemiológica da toxoplasmose no Município de Londrina-PR.The aims of this study were to verify the presence of Toxoplasma gondii cysts in fresh pork sausage and the presence of antibodies against T. gondii in serum of workers from factories with Municipal Inspection Service, in Londrina, PR, Brazil. 149 samples of sausage were collected from eight factories and blood samples from 47 workers. We also took information about the practices that were adopted in the factories and the workers' habits that could influence the prevalence of toxoplasmosis. After bioassay in mice, 13 (8.7%) sausage samples were positive, in one of them T. gondii was isolated and in the other 12 the mice seroconverted. Of 47 workers, 36 (76.6%) worked in sausage production and 11 (23.4%) were involved in other functions; 59.5% (28/47), 55.5% (20/36) and 72.7% (8/11), respectively, had T. gondii antibodies. There were no significant differences in the variables of industries' practices and workers' habits related to T. gondii infection. We concluded that fresh pork sausage could be important in the transmission of toxoplasmosis

Ocorrência de anticorpos anti-Toxoplasma gondii em caprinos de Pitanga, Paraná, Brasil

The aim of this study was to verify the prevalence of anti-Toxoplasma gondii antibodies in sera from naturally infected milk-producing caprines using indirect immunofluorescence assay (IFA) and modified agglutination test (MAT), determine specificity and sensibility of MAT and to evaluate the agreement of results between the tests. For this study, 282 caprine serum samples were collected from farms in the city of Pitanga, Paraná State, a region with a high pecuary of small ruminants. Prevalence found by IFA was of 44.68% and 23.05% by MAT. There was statistically significant difference (p=0.0086) in relation to the gender of animals, being females more seropositive. Sensitivity and specificity results for MAT were, respectively, 42.2% and 94.9%. By comparing results, a moderate agreement was observed (Kappa = 0.42). The high prevalence for the infection, associated to high titers found by IFA and clinical signs presented in the herds suggest toxoplasmosis as one of the causes of reproductive problems in caprines. Sensitivity and specificity results for MAT obtained in this study allow this test to be recommended as a confirmatory test for the caprine species.Objetivou-se verificar a prevalência de anticorpos anti-Toxoplasma gondii em soros de caprinos de produção leiteira naturalmente infectados utilizando os testes de imunofluorescência indireta (IFI) e o de aglutinação modificada (MAT), determinar a especificidade e a sensibilidade do MAT e avaliar a concordância dos resultados entre os testes. Para o estudo foram coletadas 282 amostras de soro caprino, de propriedades do município de Pitanga, Paraná, região com uma grande pecuária de pequenos ruminantes. A prevalência encontrada pela IFI foi de 44,68% e 23,05% pelo MAT. Houve diferença estatisticamente significativa (p=0,0086) em relação ao sexo dos animais, sendo que as fêmeas foram mais soropositivas. Os resultados de sensibilidade e especificidade para o MAT foram, respectivamente, 42,2% e 94,9%. Na comparação dos resultados foi observada uma concordância moderada (Kappa = 0,42). A alta prevalência da infecção, aliada aos altos títulos obtidos pela IFI e aos sinais clínicos presentes nos rebanhos sugere a toxoplasmose como uma possível causa de problemas reprodutivos em caprinos. Os resultados de sensibilidade e especificidade para o MAT obtidos neste estudo permitem recomendá-lo como teste confirmatório para a espécie caprina

Toxoplasma gondii: I. Avaliação da virulência de oito amostras

In order to evaluate the virulence and pathogenicity of tachyzoites, eight Toxoplasma gondii strains isolated from different animal species and humans - LIV IV, LIV V e S 11 isolated from swine, RH e VPS from humans, AS 28 from mice, HV III from dog and CN from cat - were inoculated in mice and rabbits. All the strains were strongly virulent for mice. Groups of mice inoculated, intraperitonially, with 10(4) tachyzoites died in an average of 6.0 to 7.8 days, after inoculation. The strains isolated more recently, LIV V and HV III (LD50 of 7 and 15 tachyzoites, respectively) were the most virulent. The RH strain showed the lowest virulence, with LD50 of 3160 tachyzoites. The LIV V strain also seems to be most virulent to rabbits.Oito amostras de T. gondii - LIV IV, LIV V e S 11 isoladas de suínos, RH e VPS de seres humanos, AS 28 de camundongo, HV III de cão e CN de gato - foram inoculadas em camundongos suíços albinos e em coelhos com o objetivo de avaliar a virulência e a patogenicidade. As oito amostras apresentaram-se altamente virulentas para camundongos, matando todos os animais que receberam inóculo, via intraperitoneal, de 10(4) taquizoítas, entre 6,0 e 7,8 dias, em média, após a inoculação. As amostras isoladas mais recentemente, LIV V e HV III (DL50 de 7 e 15 taquizoítas, respectivamente) foram as mais virulentas. A amostra RH foi a que apresentou a menor virulência, com DL50 de 3.160 taquizoítas. A amostra LIV V também se mostrou mais virulenta para coelhos, porém, como foram inoculados apenas 2 animais, estudos posteriores devem ser realizados para confirmar este achado

Seroprevalence of feline leishmaniasis in Paraná using antigens from different species of Leishmania spp / Soroprevalência da leishmaniose felina no Paraná utilizando-se antígenos de diferentes espécies de Leishmania spp

In domestic cats, the number of infections caused by parasites of the genus Leishmania is becoming increasingly significant, although the role of felines in this cycle is not yet clear. The aim of this study was to observe the prevalence of anti-Leishmania spp. using different antigens: Leishmania Viannia brasiliensis, Leishmania Leishmania amazonensis and Leishmania Leishmania infantum, in serological samples from domestic cats, from castration efforts carried out in the city of Londrina / PR, from 2015 to 2016. The test was used in indirect ELISA (Enzyme-Linked Immunosorbent Assay) serological diagnosis, with the aforementioned antigens for plaque sensitization. Of the total of 199 samples analyzed, 69 (34.67%) were considered reagents for Leishmania spp., 37 samples (18.59%) were considered reagents for L. (L.) amazonensis; 40 samples (20.10%) for L. (V.) brasiliensis and 24 (12.06%) for L. (L.) infantum. In 42 samples (21.10%) reactivity to ELISA was observed in more than one of the antigens used. A higher prevalence of antibodies was observed when using the L. (V.) brasiliensis antigen, confirming that this is the most reported species in the state of Paraná. We can conclude that antibodies against Leishmania spp. are circulating in the studied feline population; in addition, the use of the Leishmania antigen species most prevalent in the study region is indicated for obtaining more reliable results.

Soroepidemiologia e variáveis ocupacionais e ambientais relacionadas à leptospirose, brucelose e toxoplasmose em trabalhadores de frigorífico do Estado do Paraná, Brasil

A leptospirose, brucelose e a toxoplasmose são zoonoses de ampla distribuição, sendo o homem participante acidental das suas cadeias epidemiológicas. O objetivo deste trabalho foi realizar levantamento soroepidemiológico e identificar variáveis ocupacionais e ambientais relacionadas a estas enfermidades em 150 trabalhadores de um frigorífico da região Norte do Paraná. Para o diagnóstico de leptospirose foi realizada a prova de soroaglutinação microscópica; para a brucelose, a prova do antígeno acidificado tamponado e do 2-mercaptoetanol e para toxoplasmose, a reação de imunofluorescência indireta. Para cada funcionário foi preenchido um questionário epidemiológico que investigou variáveis ocupacionais e ambientais que poderiam estar associadas a estas infecções. Resultados positivos para leptospirose foram encontrados em 4,00% das amostras, para brucelose em 0,66% das amostras e para toxoplasmose 70,00%. Das três enfermidades pesquisadas, somente os resultados obtidos para leptospirose sugerem infecção de modo ocupacional.Leptospirosis, brucellosis and toxoplasmosis are widely-distributed zoonosis, being the man an accidental participant of their epidemiological chains. The aim of this paper was to make a seroepidemiological report and identify occupational and environmental variables related to these illnesses in 150 workers in a slaughterhouse in the Northern region of Paraná. For the diagnosis of leptospirosis a microscopical seroagglutination test was applied; for brucellosis, the tamponated acidified antigen test and the 2-mercaptoetanol tests were used, and for toxoplasmosis the indirect immunofluorescence reaction test. For each employee an epidemiological survey was filled, which investigated occupational and environmental variables which could be associated with these infections. Positive results for leptospirosis were found in 4.00% of the samples, for brucellosis in 0.66% of samples and toxoplasmosis in 70.00%. From the three diseases researched, only the results for leptospirosis suggest occupational infection

"Baixadeiros" horses: prevalence of anti-Trypanosoma spp. and anti-Leishmania spp. antibodies

A total of 138 horse blood samples were collected from 2012 to 2013 for the subsequent separation of serum collection. For detection of anti-Leishmania spp. and anti-Trypanosoma spp. antibodies, indirect immunofluorescence reactions and immunoenzymatic assays were performed. Samples with titers ≥40 were considered positive in the indirect immunofluorescence (IFAT), and the cut off for the enzyme immunoassay (ELISA) was calculated with three standard deviations. When tested for Trypanosoma spp., 50.37% (68/135) of the samples were ELISA reagents, 18.84% (8/136) were positive for IFAT and 2.89% (4/138) for both tests. A significant statistical association was observed for the municipality (p = 0.013) and collection period (p = 0.042) in the ELISA. When tested for Leishmania spp., 25.4% (35/138) of the samples were ELISA reagents, 13.00% (18/138) were positive for the IFAT and 4.34% (6/138) were positive for two tests, there were no variables associated with seropositivity. A statistical association (p = 0.0034) was observed between the presence of anti-Trypanosoma spp. and anti-Leishmania spp. antibodies when the ELISA was used. The results suggest that this horse breed is in contact with leishmaniasis and trypanosomiasis, which demonstrates the need for a more accurate investigation into the real role of horses in these diseases to assist in disease control measures.Keywords: Leishmaniasis; Trypanosomiasis; Equines; Serolog

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% 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.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% de gestantes IgG reagente e IgM não reagente, 1,9% de IgG e IgM reagentes, 0,3% de IgG não reagente e IgM reagente e 44,7% de IgG e IgM não reagentes. No grupo II foram 55,1% de IgG reagente e IgM não reagente e 44,9% 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

Toxoplasma gondii: II. Tachyzoite antigenic characterization of eigth strains

Oito amostras de T. gondii - LIV IV, LIV V e S 11 isoladas de suínos, RH e VPS de seres humanos, AS 28 de camundongo, HV III de cão e CN de gato - foram analisadas com o objetivo de verificar a existência de possíveis diferenças na resposta imune quando inoculadas em coelhos. Através da técnica de ELISA, não foram constatadas diferenças entre as oito amostras estudadas. Todas as amostras reagiram de forma semelhante com soros homólogos e heterólogos. A suspensão antigênica, constituída de extrato celular total, mostrou-se eficiente no ELISA teste indireto, já que os soros positivos reagiram fortemente e os soros negativos não apresentaram reação contra os antígenos testados. A análise das amostras, pela técnica de Western blot, revelou que os isolados de T. gondii compartilham vários antígenos com algumas variações. Dentre as bandas reconhecidas no Western blot, três foram comuns a todas as amostras: a p33 (33-37 kDa), p54 (52-55 kDa) e a p66 (66 kDa). A amostra HV III, isolada recentemente de um cão, foi a que mais diferiu no perfil antigênico. Essa amostra não apresentou três antígenos (50, 70 e 75 kDa) presentes nas demais amostras. Apenas dois antígenos, um de 62 kDa da CN e outro de 67 kDa da LIV IV, foram amostra-específicos.Eight Toxoplasma gondii strains were analyzed using ELISA and Western blot techniques, in order to demonstrate possible immunological differences. The analyzed strains were: LIV IV, LIV V and S 11 isolated from swine, RH and VPS from a human being, AS 28 from a wild mouse, HV III from a dog and CN from a cat. With the ELISA assay the eight strains showed similar reactivity with homologous and heterologous sera. The antigenic suspension, consisting of total cellular extract of tachyzoites, was effective in the indirect ELISA assay, with the positive sera reacting strongly and the negative not reacting with the antigens. The Western blot analysis showed that the T. gondii strains have similar antigenic profiles with a few variations. Three bands were observed in all strains: one of about 33 kDa (p33), another of 54 kDa (p54) and a third one of 66 kDa (p66). The HV III strain, isolated from a dog, did not show three antigens (50, 70 and 75 kDa) that were present in the others. However, this difference was not detected by the ELISA assay. Only two antigens (62 kDa of the CN and 67 kDa of the LIV IV) were strain-specific antigens

Cross-Sectional Study of Leptospira spp. and Brucela abortus in Goat Herds from Paraná State, Brazil

Background: Leptospirosis and brucellosis are zoonosis worldwide distributed that have great economic importance, especially in goats. This study aimed to carry out a cross-sectional study to determine the prevalence of anti-Leptospira spp. and anti-Brucella abortus antibodies and to determine epidemiological variables associated to infection by these ethiological agents in goats.Materials, Methods & Results: Serum samples were collected from 1055 goats of 95 properties distributed in 18 regional centers of Paraná State from April to August of 2010. Collected samples were submitted to microscopic agglutination test (MAT) for the detection of anti-Leptospira spp. antibodies with 22 reference serovars: Australis, Bratislava, Autumnalis, Butembo, Castellonis, Bataviae, Canicola, Fortbragg, Whitcombi, Cynopteri, Grippotyphosa, Hebdomadis, Copenhageni, Icterohaemorrhagiae, Panama, Pomona, Pyrogenes, Hardjo, Wolffi, Shermani, Tarassovi and Londrina 1 (L1). The samples were also submitted to buffered acidified plate antigen test (BAPA) and 2-mercaptoethanol test (2-ME) to detect anti-Brucella abortus antibodies. The results obtained from epidemiological and serological study were analysed by Epi Info 3.5.3. We observed 9.9% (104/1055) of goats reagents in at least one serovar of Leptospira spp. and no reagent animal to Brucella abortus. The variables that were statistically significant with leptospirosis were: production type meat, sewer destination in dry sump, frequent abortion, shared use of pastures, slaughter of animals on the property, the presence of pigs and wild animals such as deer and capybaras on the property.Discussion: This was the first study about antibodies prevalence against Leptospira spp. and Brucella abortus in goat herds from Paraná State. The presence of both animal species were statiscally associated to risk of leptospirosis. It is necessary avoid the acess of these animals in herds, decreasing the dissemination of the agent. The production type meat is a form of creation in which the animals have no care of zookeepers and continuous veterinary care. The final destination of sewage was statistically associated to risk of leptospirosis in goats and this is a big problem in rural properties, requiring greater attention of authorities for proper disposal of this wastewater, reducing water and food contamination. Among the variables related to management, the herds that used rented pasture or shared pastures also were statistically associated to risk of leptospirosis in goats; with the sharing of pastures a greater flow of animals transiting in the pickets occurs, allowing not only flocks of goats, but other species also, to use the same pastures and become more exposed to certain diseases. The slaughter of pigs, sheep, cattle and goats on the property is a problem for animal and human health due to lack of sanitary inspection. Abortion is one of the main clinical signs of leptospirosis in farm animals, particularly goats. The knowledge of these results will be useful to develop control programs for the disease in Paraná State herds.  The absence of circulating anti-Brucella abortus antibodies in goat herds has also been reported in other Brazilian States