João Luis Garcia

Soroprevalência do toxoplasma gondii, em suínos, bovinos, ovinos e eqüinos, e sua correlação com humanos, felinos e caninos, oriundos de propriedades rurais do norte do Paraná-Brasil

Toxoplasma gondii the agent causing toxoplasmosis has a worldwide distribution. In the present study we attempt to determine the prevalence of toxoplasmosis in food animals and horses from farms in Jaguapita county. IgG antibody were detected using indirect immunofluorescence. Titers higher or equal to 1:64 were considered reactive. We found prevalence rates of 24% (267) for swine, 25.8% (400) for bovine , 51.8% (228) for ovine samples, and 12.1% (173) for equine. Seropositivity increased with age in sheeps and pigs, there were no gender differences in the species studied. There was a positive and significant correlation between human and feline (r= 0.64 p=0.05), human and canine (r=0.78 p=0.01), canine and swine (r=0.96 p=0.0001), bovine and ovine (r=0.82 p=0.006), bovine and equine (r=0.89 p=0.001), and ovine and equine (r=0.92 p=0.0004) titers. These results suggest that carnivores, herbivores, and omnivores are exposed to the same sources of transmission. Toxoplasmosis has a high prevalence on the studied population. Our results suggest that raw or rare cooked meat may an important way of transmiting toxoplasmosis to humans in this region.

Soroprevalência, epidemiologia e avaliação ocular da toxoplasmose humana na zona rural de Jaguapitã (Paraná), Brasil

A toxoplasmose e uma protozoonose de distribuicao mundial, que pode infectar uma grande variedade de especies. No presente estudo, avaliou-se a presenca de toxoplasmose em 345 moradores da zona rural do Municipio de Jaguapita, Estado do Parana, Brasil. A frequencia dos titulos nas amostras de soro humano foi comparada com a frequencia dos titulos encontrados em 1 420 amostras de diversas especies animais com as quais os moradores da regiao tinham contato. A prevalencia de anticorpos anti-Toxoplasma gondii foi de 66% entre a populacao do estudo (reacao de imunofluorescencia indireta). Foram considerados positivos os titulos maiores ou iguais a 16. O maior titulo encontrado foi de 65 536 (1%) e os titulos mais frequentes foram de 256 (29%) e 1024 (19%). A comparacao entre seres humanos e animais revelou uma correlacao positiva e significativa entre a distribuicao dos titulos de anticorpos de humanos e felinos (r = 0,78; P = 0,01) e de humanos e caninos (r = 0,64; P= 0,05). Os participantes do estudo foram tambem testados pela tela de Amsler, sendo que 22% deles (75/345) relataram algum tipo de alteracao ocular; destas pessoas, 77% (58/75) foram sororreagentes a toxoplasmose. Quarenta e uma das 58 pessoas foram encaminhadas para exame oftalmico de fundo de olho. Neste grupo, 22% dos pacientes (9/41) apresentaram lesoes caracteristicas de coriorretinite cicatrizada, sugestivas de toxoplasmose ocular (idade entre 34 e 78 anos). Nenhum paciente apresentou processo inflamatorio. Dentre seis pacientes (67%) com lesoes unilaterais, quatro apresentaram titulo de anticorpos de 256. O inquerito epidemiologico revelou que os individuos sororreagentes tinham 2,06 vezes mais possibilidades de apresentarem problemas oculares do que os nao reagentes. Neste sentido, nao foram verificadas diferencas significativas com relacao a sexo, contato com animais, consumo de carne crua ou mal passada e leite cru e abate de animais para consumo proprio. Os resultados sugerem que a toxoplasmose esta amplamente distribuida na regiao, com significativa incidencia de lesoes oculares provocadas pelo Toxoplasma gondii. E recomendavel um maior controle por parte das autoridades sanitarias, visando diminuir o risco de infecao toxoplasmica, principalmente entre gestantes.

Soroepidemiologia da toxoplasmose em gatos e cães de propriedades rurais do município de Jaguapitã, estado do Paraná, Brasil

The present study was designed to stablish the seroprevalence of toxoplasmosis in cats and dogs from a rural area of the Jaguapita county, Parana state, Antibodies against-Toxoplasma gondii were detected by indirect immuno-fluorescence test. Titers than 1:16 were considered positive. A prevalence rate of 73% and 84.1% was observed for cats and dogs respectively . There was no difference related to sender. The most frequent titer in cats was 1:4096 (28.6%), whereas the highest titer found was 1:65536 (0.8%). There was a statistical significant increase in seropositivity with age. The most common titers in dogs was 1:64 (38.4%), whereas the highest titer was 1:4096 (1.9%). There was a decrease in seropositivity in dogs with less than 8 of age. There was no significant correlation in the distribuition of titers between dogs and cats. Based on our data we conclude that T. gondii is widely distributed in dogs and cats from this region.

Factors associated with seropositivity for anti-Toxoplasma gondii antibodies in pregnant women of Londrina, Paraná, Brazil

The aim of this study was to evaluate associations between seropositivity for IgG and IgM anti-Toxoplasma gondii antibodies and socio-economic and environmental variables in pregnant women of Londrina, state of Paraná, Brazil. We interviewed 492 pregnant women, each of whom answered an epidemiological questionnaire, and collected blood samples for measurement of IgG and IgM anti-T. gondii antibodies by chemiluminescence. A confirmatory diagnosis of acute infection was made by an IgG avidity test. Titres of specific IgG anti-T. gondii were obtained by IFAT. Seropositivity for IgG anti-T. gondii antibodies was observed in 242 women (49.2%) and, of these, six pregnant women (1.2%) showed seropositivity for IgM. Age group, level of education, per capita income, presence of a cat in the house and a habit of eating green vegetables were all factors associated with a greater chance of infection with T. gondii. This study showed that 250 (50.8%) pregnant women were susceptible to T. gondii and considered to be at high risk for toxoplasmosis during pregnancy. Based on the results obtained, is critical to establish a program of health surveillance for toxoplasmosis, in order to contribute to diagnosis and early treatment during the prenatal period. It is also necessary to introduce measures to prevent the Toxoplasma infection in seronegative pregnant women.

SEROPREVALENCE OF Toxoplasma gondii IN BACK YARD CHICKEN (Gallus gallus domesticus) FROM RURAL PROPERTIES IN NORTH REGION OF PARANÁ STATE, BRAZIL

O Toxoplasma gondii e um protozoario que pode infectar todos os animais homeotermicos. No presente trabalho, verificou-se a ocorrencia de anticorpos anti-T. gondii em galinhas de criacoes domesticas (fundo de quintal), oriundas de propriedades rurais localizadas no municipio de Jaguapita, Estado do Parana, Brasil. Estudos previos demostraram que o parasita esta amplamente disseminado nas diferentes especies de animais domesticos, de companhia e no homem nessas mesmas propriedades. Os soros foram submetidos a reacao de Imunofluorescencia Indireta, sendo a positividade considerada para aqueles titulos maiores ou iguais a 16. Foram estudados 155 soros , sendo que 16 (10,3%) foram reagentes a toxoplasmose, e 139 (89,7%) nao reagentes. Os titulos mais frequentes foram de 64 (5/31,2%), 16 e 1024 (ambos com 4/25%) e o maior titulo encontrado foi de 1024 (4/25%). A soroprevalencia do T. gondii nao foi influenciada pelo sexo da ave (Exato de Fisher p = 0,12), pela finalidade da criacao (corte ou postura, Exato de Fisher p = 0,39), pela percentagem de felinos sororeagentes na propriedade (r = -0,19 p = 0,27), nem pelo numero de felinos sororeagentes (r = -0,09 p = 0,60). Atraves do presente trabalho, demostrou-se uma ocorrencia elevada da resposta sorologica ao T. gondii em aves domesticas oriundas de propriedades rurais neste municipio, consequentemente esses animais podem representar risco de infeccao para o homem e para outros animais quando consumidos crus ou mal cozidos.

Occurrence of Ehrlichia canis and Anaplasma platys in household dogs from northern Parana

Canine monocytic ehrlichiosis caused primarily by Ehrlichia canis and canine thrombocytic anaplasmosis induced by Anaplasma platys are important emerging zoonotic tick-borne diseases of dogs. There is evidence that these pathogens can also affect humans. This study evaluated the presence of E. canis and A. platys in blood samples collected from 256 domiciled dogs in the municipality of Jataizinho, located in north region of the State of Parana, Brazil, by PCR assay. The occurrence of E. canis and A. platys was 16.4% (42/256) and 19.4% (49/256), respectively; while 5.47% (14/256) of the dogs evaluated were co-infected by these two organisms. The presence of E. canis and A. platys was not significantly associated with the variables evaluated (sex, age, outdoor access, and presence of ticks during blood collection). Infection of dogs by E. canis was associated with anemia and thrombocytopenia, while infection induced by A. platys was related only to thrombocytopenia. Canine monocytic ehrlichiosis and canine thrombocytic anaplasmosis should be included in the differential diagnoses when these hematological alterations are observed during routine laboratory evaluation of dogs.

Soroepidemiologia da toxoplasmose e avaliação ocular pela Tela de Amsler, em pacientes da zona rural, atendidos na unidade de saúde do município de Jaguapitã, PR, Brasil

In this study, we collected serum from 82 randomly selected patients from a rural area in Jaguapitã county, Paraná State. All sera were subjected to indirect immunofluorescence assay (IFA) to detect IgG antibodies against T. gondii. Serum titers >/=+ 16 were considered positive. Of the 82 evaluated sera, 68 (82.9%) were positive to toxoplasmosis and 14 (17.1%) were non-reactive. The most common titers found were 64 (23/33.8%) and 256 (16/23.5%), the highest titer being 4096 (8/11.8%). The 82 patients were also evaluated by the Amsler Grid test, with 22 (26.8%) patients presenting at least one alteration in this test. Masculinity was found to be a protection factor in the Amsler grid test (OR = 0.21 0.04 < OR < 0. 86 chi2 = 4,98 p = 0,02). There were no significant statistical differences related to the epidemiological investigation. We concluded that T. gondii has a wide distribution in the population studied.

Vaccination of pigs with the S48 strain of Toxoplasma gondii - safer meat for human consumption

As clinical toxoplasmosis is not considered a problem in pigs, the main reason to implement a control strategy against Toxoplasma gondii (T. gondii) in this species is to reduce the establishment of T. gondii tissue cysts in pork, consequently reducing the risk of the parasite entering the human food chain. Consumption of T. gondii tissue cysts from raw or undercooked meat is one of the main sources of human infection, with infected pork being considered a high risk. This study incorporates a mouse bioassay with molecular detection of T. gondii DNA to study the effectiveness of vaccination (incomplete S48 strain) in its ability to reduce tissue cyst burden in pigs, following oocyst (M4 strain) challenge. Results from the mouse bioassay show that 100% of mice which had received porcine tissues from vaccinated and challenged pigs survived compared with 51.1% of mice which received tissues from non-vaccinated and challenged pigs. The presence (or absence) of T. gondii DNA from individual mouse brains also confirmed these results. This indicates a reduction in viable T. gondii tissue cysts within tissues from pigs which have been previously vaccinated with the S48 strain. In addition, the study demonstrated that the main predilection sites for the parasite were found to be brain and highly vascular muscles (such as tongue, diaphragm, heart and masseter) of pigs, while meat cuts used as human food such as chop, loin, left tricep and left semitendinosus, had a lower burden of T. gondii tissue cysts. These promising results highlight the potential of S48 strain tachyzoites for reducing the number of T. gondii tissues cysts in pork and thus improving food safety.

Diagnosis and isolation of Toxoplasma gondii in horses from Brazilian slaughterhouses

This study aimed to investigate anti-Toxoplasma gondii antibodies and to isolate the parasite from the brains of horses processed at slaughterhouses in Brazil. We collected brain and blood samples from 398 horses of various ages, from six Brazilian states. Serum samples were evaluated by indirect fluorescent antibody test (IFAT cut-off titre ≥ 1:64), and brains were submitted to mouse bioassay. Among the 398 horses, positivity for T. gondii was identified in 46 (11.6%) by IFAT and in 14 (3.5%) by mouse bioassay. In 12 of those 14 bioassays, mice were positive only by IFAT (cut-off titre ≥ 1:16), T. gondii being isolated in the remaining two. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of 18S rDNA to differentiate among T. gondii, Neospora caninum, and Sarcocystis neurona, we found that two of the 14 brains were positive for T. gondii only. For genotyping of the two isolates and the PCR-positive brain, we performed PCR-RFLP based on 13 markers, and SAG2 all samples were Toxoplasma gondii type I. Collectively, IFAT of horse sera and mouse bioassay identified positivity in 60 (15%) of the samples. Our results show that some horses sent to slaughter in Brazil have been exposed to T. gondii.

Concomitant canine distemper, infectious canine hepatitis, canine parvoviral enteritis, canine infectious tracheobronchitis, and toxoplasmosis in a puppy

The concomitant infections of Canine distemper virus (CDV), Canine adenovirus A types 1 (CAdV-1) and 2 (CAdV-2), Canine parvovirus type 2 (CPV-2), and Toxoplasma gondii are described in a 43-day-old mixed-breed puppy. Clinically, there were convulsions and blindness with spontaneous death; 14 siblings of this puppy, born to a 10-month-old dam, which was seropositive (titer: 1,024) for T. gondii, also died. Necropsy revealed unilateral corneal edema (blue eye), depletion of intestinal lymphoid tissue, non-collapsible lungs, congestion of meningeal vessels, and a pale area in the myocardium. Histopathology demonstrated necrotizing myocarditis associated with intralesional apicomplexan protozoa; necrotizing and chronic hepatitis associated with rare intranuclear inclusion bodies within hepatocytes; necrotizing bronchitis and bronchiolitis; interstitial pneumonia associated with eosinophilic intracytoplasmic inclusion bodies within epithelial cells; atrophy and fusion of intestinal villi with cryptal necrosis; and white matter demyelination of the cerebrum and cerebellum associated with intranuclear inclusion bodies within astrocytes. Polymerase chain reaction (PCR) amplified the partial fragments (bp) of the CDV N gene (290 bp), CPV-2c VP2 capsid protein gene (583 bp), and CAdV-1 (508 bp) and CAdV-2 (1,030 bp) E gene from urine and tissue samples. The PCR assays demonstrated that the apicomplexan protozoa observed within several organs contained DNA specific for T. gondii; genotyping revealed T. gondii type III. The findings support the characterization of concomitant infections of CDV, CAdV-1, CAdV-2, CPV-2, and T. gondii in this puppy. Further, seroreactivity to T. gondii of the dam in association with the systemic disease observed in the puppy described herein is suggestive of congenital toxoplasmosis.