Ocorrência de anticorpos anti-Toxoplasma gondii e anti-Neospora caninum em ovinos com distúrbios reprodutivos e fatores de risco / Occurrence of antibodies anti-Toxoplasma gondii and anti-Neospora caninum in sheep with history of reproductive disorders and risk factors

Objetivou-se avaliar a ocorrência, sinais clínicos e fatores de risco associados a soropositividade para Toxoplasma gondii e Neospora caninum em ovinos. Foram utilizados 294 animais com histórico de distúrbios reprodutivos de 28 fazendas do estado de São Paulo, Brasil, diagnosticados através da imunofluorescência indireta (1:64 e 1:50). A ocorrência de T. gondii foi de 29,9% (88/294) e de N. caninum 18% (53/294), sendo 3,7% (11/294) dos ovinos soropositivos para ambos. Observou-se com maiores chances à infecção pelo T. gondii: ovinos mestiços (p=0,04), Santa Inês (p=0,006), fornecimento de pastagem (p<0,001) ou associada a concentrado (p<0,001), uso exclusivamente de monta natural (p=0,002, OR=2,28 e IC95%=1,37-3,79) e a presença de aves nas propriedades (p=0,001). Na infecção por N. caninum essa chance aumentou em: fêmeas (p=0,031), propriedades sem aprisco (p<0,001) e sistema de criação semi-intensivo (p<0,001). Em relação ao histórico de problemas reprodutivos, ovelhas infectadas pelo N. caninum e T. gondii apresentaram redução da chance de apresentarem abortamento (p=0,044) e repetição de estro (p=0,025) respectivamente. O T. gondii esteve mais presente sorologicamente que o N. caninum em ovinos com histórico de distúrbios reprodutivos e apesar de suas semelhanças, diferiram epidemiologicamente em aspectos relacionados as características da criação como raça, sexo, sistema de criação, tipo de alimentação e manejo reprodutivo.(AU)

The objective was to evaluate the occurrence, clinical signs and risk factors associated with seropositivity to Toxoplasma gondii and Neospora caninum in sheep. We used 294 sheep with history of reproductive disorders from 28 farms located in the state of São Paulo, southeastern Brazil. Producers were interviewed, and blood samples were collected to perform indirect immunofluorescence tests (1:64 and 1:50 respectively). The frequency of T. gondii infection was found to be 29.9% (88/294), the frequency of N. caninum was 18% (53/294), and 3.7% (11/294) of the sheep were seropositive for both. We observed the following risk factors associated with T. gondii infection: crossbred sheep (p=0.04), Santa Inês breed (p=0.006), pasture supply (p<0.001) or associated with concentrate (p<0.001), exclusive use of natural breeding (p=0.002), and presence of birds on farms (p=0.001). For N. caninum the factors were: female sheep (p=0.031), absence of barns (p<0.001), and semi-intensive system (p<0.001). In relation to the history of reproductive problems, N. caninum and T. gondii infected sheep presented a reduction in the risk of having an abortion (p=0.044) and repeated estrus (p=0.025) respectively. T. gondii was more serologically present than N. caninum in sheep with a history of reproductive disorders and, despite their similarities, differed epidemiologically in aspects related to breeding characteristics such as race, sex, breeding system, type of feeding and reproductive management.(AU)

Isolation and Genotyping of Toxoplasma gondii Strains in Ovine Aborted Fetuses in Khorasan Razavi Province, Iran

Toxoplasmosis is an important zoonotic disease that can cause abortion in humans and animals. The aim of this study was isolation and subsequent genotyping of Toxoplasma gondii isolates in ovine aborted fetuses. During 2012-2013, 39 ovine aborted fetuses were collected from sheep flocks in Khorasan Razavi Province, Iran. The brain samples were screened for detection of the parasite DNA by nested PCR. The positive brain samples were bioassayed in Webster Swiss mice. The serum samples of mice were examined for T. gondii antibodies by IFAT at 6 weeks post inoculation, and T. gondii cysts were searched in brain tissue samples of seropositive mice. The positive samples were genotyped by using a PCR-RLFP method. Subsequently, GRA6 sequences of isolates were analyzed using a phylogenetic method. The results revealed that T. gondii DNA was detected in 54% (20/37, 95% CI 38.4-69.0%) brain samples of ovine aborted fetuses. In bioassay of mice, only 2 samples were virulent and the mice were killed at 30 days post inoculation, while the others were non-virulent to mice. The size of cysts ranged 7-22 µm. Complete genotyping data for GRA6 locus were observed in 5 of the 20 samples. PCR-RLFP results and phylogenetic analysis revealed that all of the isolated samples were closely related to type I. For the first time, we could genotype and report T. gondii isolates from ovine aborted fetuses in Khorasan Razavi Province, Iran. The results indicate that the T. gondii isolates are genetically related to type I, although most of them were non-virulent for mice.

MOLECULAR CHARACTERIZATION AND SEQUENCE PHYLOGENETIC ANALYSIS OF SURFACE ANTIGEN 3 (SAG3) GENE OF LOCAL INDIAN ISOLATES (CHENNAI AND IZATNAGAR) OF Toxoplasma gondii / Caracterização molecular e análise filogenética de sequências do antígeno de superfície 3 (SAG3) em isolados indianos (CHENNAI E IZATNAGAR) de Toxoplasma gondii

Context and objective: The molecular characterization of local isolates of Toxoplasma gondii is considered significant so as to assess the homologous variations between the different loci of various strains of parasites. Design and setting: The present communication deals with the molecular cloning and sequence analysis of the 1158 bp entire open reading frame (ORF) of surface antigen 3 (SAG3) of two Indian T. gondii isolates (Chennai and Izatnagar) being maintained as cryostock at the IVRI. Method: The surface antigen 3 (SAG3) of two local Indian isolates were cloned and sequenced before being compared with the available published sequences. Results: The sequence comparison analysis revealed 99.9% homology with the standard published RH strain sequence of T. gondii. The strains were also compared with other established published sequences and found to be most related to the P-Br strain and CEP strain (both 99.3%), and least with PRU strain (98.4%). However, the two Indian isolates had 100% homology between them. Conclusion: Finally, it was concluded that the Indian isolates were closer to the RH strain than to the P-Br strain (Brazilian strain), the CEP strain and the PRU strains (USA), with respect to nucleotide homology. The two Indian isolates used in the present study are known to vary between themselves, as far as homologies related to other genes are concerned, but they were found to be 100% homologous as far as SAG3 locus is concerned. This could be attributed to the fact that this SAG3 might be a conserved locus and thereby, further detailed studies are thereby warranted to exploit the use of this particular molecule in diagnostics and immunoprophylactics. The findings are important from the point of view of molecular phylogeny. .

Contexto e objetivo. A caracterização molecular de isolados indianos de Toxoplasma gondii é importante para a investigação de variações genéticas existentes entre cepas do parasito em diferentes locos gênicos. Delineamento e disposição. A presente comunicação realizou a clonagem e o sequenciamento dos 1158 pares de base correspondendo à totalidade do quadro de leitura do antígeno de superfície 3 (SAG3) de Toxoplasma gondii em dois isolados indianos (Chennai e Izatnagar) mantidos em um biorrepositório localizado em IVRI. Método. As sequências do SAG3 dos dois isolados indianos foram clonadas, sequenciadas e posteriormente comparadas com sequências SAG3 de Toxoplasma gondii disponíveis em publicações. Resultados. A comparação das sequências revelou 99,9% de homologia com a cepa RH padrão; 99,3% de homologia com as cepas P-Br e CEP; e 98,4% de homologia com a cepa PRU. Os dois isolados indianos eram 100% idênticos no que diz respeito à sequência SAG3. Conclusão. Concluiu-se que os isolados indianos são filogeneticamente mais próximos da cepa RH em relação à cepa brasileira P-Br, ou às cepas CEP e PRU (USA). No entanto, a análise de outros genes de Toxoplasma gondii destes dois isolados indianos mostrou diferenças na composição de nucleotídeos, ao contrário do que foi encontrado para o locus SAG3. Estes resultados poderiam ser atribuídos ao fato do locus SAG3 ser altamente conservado, necessitando de estudos adicionais para determinar se SAG3 poderia ser utilizado no diagnóstico da toxoplasmose. No entanto, estes resultados são importantes do ponto de vista da filogenia molecular. .

Prevalence of Toxoplasma gondii in Dogs in Zhanjiang, Southern China

Toxoplasmosis, caused by Toxoplasma gondii, is a parasitic zoonosis with worldwide distribution. The present study investigated the prevalence of T. gondii in dogs in Zhanjiang city, southern China, using both serological and molecular detection. A total of 364 serum samples and 432 liver tissue samples were collected from the slaughter house between December 2012 and January 2013 and were examined for T. gondii IgG antibody by ELISA and T. gondii DNA by semi-nested PCR based on B1 gene, respectively. The overall seroprevalence of T. gondii IgG antibody was 51.9%, and T. gondii DNA was detected in 37 of 432 (8.6%) liver tissue samples. These positive DNA samples were analyzed by PCR-RFLP at 3'- and 5'-SAG2. Only 8 samples gave the PCR-RFLP data, and they were all classified as type I, which may suggest that the T. gondii isolates from dogs in Zhanjiang city may represent type I or type I variant. This study revealed the high prevalence of T. gondii infection in dogs in Zhanjiang city, southern China. Integrated measures should be taken to prevent and control toxoplasmosis in dogs in this area for public health concern.

Genetic Diversity of Toxoplasma gondii Strains from Different Hosts and Geographical Regions by Sequence Analysis of GRA20 Gene

Toxoplasma gondii is a eukaryotic parasite of the phylum Apicomplexa, which infects all warm-blood animals, including humans. In the present study, we examined sequence variation in dense granule 20 (GRA20) genes among T. gondii isolates collected from different hosts and geographical regions worldwide. The complete GRA20 genes were amplified from 16 T. gondii isolates using PCR, sequence were analyzed, and phylogenetic reconstruction was analyzed by maximum parsimony (MP) and maximum likelihood (ML) methods. The results showed that the complete GRA20 gene sequence was 1,586 bp in length among all the isolates used in this study, and the sequence variations in nucleotides were 0-7.9% among all strains. However, removing the type III strains (CTG, VEG), the sequence variations became very low, only 0-0.7%. These results indicated that the GRA20 sequence in type III was more divergence. Phylogenetic analysis of GRA20 sequences using MP and ML methods can differentiate 2 major clonal lineage types (type I and type III) into their respective clusters, indicating the GRA20 gene may represent a novel genetic marker for intraspecific phylogenetic analyses of T. gondii.

Sequence Diversity in MIC6 Gene among Toxoplasma gondii Isolates from Different Hosts and Geographical Locations

Toxoplasma gondii is an opportunistic protozoan parasite that can infect almost all warm-blooded animals including humans with a worldwide distribution. Micronemes play an important role in invasion process of T. gondii, associated with the attachment, motility, and host cell recognition. In this research, sequence diversity in microneme protein 6 (MIC6) gene among 16 T. gondii isolates from different hosts and geographical regions and 1 reference strain was examined. The results showed that the sequence of all the examined T. gondii strains was 1,050 bp in length, and their A + T content was between 45.7% and 46.1%. Sequence analysis presented 33 nucleotide mutation positions (0-1.1%), resulting in 23 amino acid substitutions (0-2.3%) aligned with T. gondii RH strain. Moreover, T. gondii strains representing the 3 classical genotypes (Type I, II, and III) were separated into different clusters based on the locus of MIC6 using phylogenetic analyses by Bayesian inference (BI), maximum parsimony (MP), and maximum likelihood (ML), but T. gondii strains belonging to ToxoDB #9 were separated into different clusters. Our results suggested that MIC6 gene is not a suitable marker for T. gondii population genetic studies.

Sequence Variation in Superoxide Dismutase Gene of Toxoplasma gondii among Various Isolates from Different Hosts and Geographical Regions

Toxoplasma gondii, an obligate intracellular protozoan parasite of the phylum Apicomplexa, can infect all warm-blooded vertebrates, including humans, livestock, and marine mammals. The aim of this study was to investigate whether superoxide dismutase (SOD) of T. gondii can be used as a new marker for genetic study or a potential vaccine candidate. The partial genome region of the SOD gene was amplified and sequenced from 10 different T. gondii isolates from different parts of the world, and all the sequences were examined by PCR-RFLP, sequence analysis, and phylogenetic reconstruction. The results showed that partial SOD gene sequences ranged from 1,702 bp to 1,712 bp and A + T contents varied from 50.1% to 51.1% among all examined isolates. Sequence alignment analysis identified total 43 variable nucleotide positions, and these results showed that 97.5% sequence similarity of SOD gene among all examined isolates. Phylogenetic analysis revealed that these SOD sequences were not an effective molecular marker for differential identification of T. gondii strains. The research demonstrated existence of low sequence variation in the SOD gene among T. gondii strains of different genotypes from different hosts and geographical regions.

Prevalence and Genetic Characterization of Toxoplasma gondii in Pet Dogs in Central China

The prevalence and genotype of Toxoplasma gondii infection in dogs in Henan Province, Central China was investigated. A total of 125 blood samples were collected from pet dogs during April to June 2013, and all samples were examined by indirect hemagglutination antibody test (IHA) and nested PCR. The overall T. gondii prevalence in pet dogs was 24.0% (30/125), with 20.8% (26/125) in IHA and 10.4% (13/125) in PCR, respectively. No statistical associations were found between animal gender and age and the prevalence of T. gondii infection. Thirteen positive DNA samples were genotyped using 11 PCR-RFLP markers, including SAG1, (3'+5') SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. Of these, only 2 samples were genotyped with complete data for all loci, and a novel genotype (type III at SAG3 and GRA6 loci, and type I at other loci) was identified. This is the first report of genetic characterization of T. gondii infection in dogs in China.

Prevalence and Genetic Characterization of Toxoplasma gondii in Pet Dogs in Central China

The prevalence and genotype of Toxoplasma gondii infection in dogs in Henan Province, Central China was investigated. A total of 125 blood samples were collected from pet dogs during April to June 2013, and all samples were examined by indirect hemagglutination antibody test (IHA) and nested PCR. The overall T. gondii prevalence in pet dogs was 24.0% (30/125), with 20.8% (26/125) in IHA and 10.4% (13/125) in PCR, respectively. No statistical associations were found between animal gender and age and the prevalence of T. gondii infection. Thirteen positive DNA samples were genotyped using 11 PCR-RFLP markers, including SAG1, (3'+5') SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. Of these, only 2 samples were genotyped with complete data for all loci, and a novel genotype (type III at SAG3 and GRA6 loci, and type I at other loci) was identified. This is the first report of genetic characterization of T. gondii infection in dogs in China.

Seropositivity and Serointensity of Toxoplasma gondii Antibodies and DNA among Patients with Schizophrenia

The aim of this cross sectional case control study was to examine the serofrequency and serointensity of Toxoplasma gondii (Tg) IgG, IgM, and DNA among patients with schizophrenia. A total of 101 patients with schizophrenia and 55 healthy controls from Sungai Buloh Hospital, Selangor, Malaysia and University Malaya Medical Center (UMMC) were included in this study. The diagnosis of schizophrenia was made based on the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV). The presence of Tg infection was examined using both indirect (ELISA) and direct (quantitative real-time PCR) detection methods by measuring Tg IgG and IgM and DNA, respectively. The serofrequency of Tg IgG antibodies (51.5%, 52/101) and DNA (32.67%, 33/101) among patients with schizophrenia was significantly higher than IgG (18.2%, 10/55) and DNA (3.64%, 2/55) of the controls (IgG, P=0.000, OD=4.8, CI=2.2-10.5; DNA, P=0.000, OD=12.9, CI=2.17-10.51). However, the Tg IgM antibody between patients with schizophrenia and controls was not significant (P>0.005). There was no significant difference (P>0.005) in both serointensity of Tg IgG and DNA between patients with schizophrenia and controls. These findings have further demonstrated the strong association between the active Tg infection and schizophrenia.

Seropositivity and Serointensity of Toxoplasma gondii Antibodies and DNA among Patients with Schizophrenia

The aim of this cross sectional case control study was to examine the serofrequency and serointensity of Toxoplasma gondii (Tg) IgG, IgM, and DNA among patients with schizophrenia. A total of 101 patients with schizophrenia and 55 healthy controls from Sungai Buloh Hospital, Selangor, Malaysia and University Malaya Medical Center (UMMC) were included in this study. The diagnosis of schizophrenia was made based on the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV). The presence of Tg infection was examined using both indirect (ELISA) and direct (quantitative real-time PCR) detection methods by measuring Tg IgG and IgM and DNA, respectively. The serofrequency of Tg IgG antibodies (51.5%, 52/101) and DNA (32.67%, 33/101) among patients with schizophrenia was significantly higher than IgG (18.2%, 10/55) and DNA (3.64%, 2/55) of the controls (IgG, P=0.000, OD=4.8, CI=2.2-10.5; DNA, P=0.000, OD=12.9, CI=2.17-10.51). However, the Tg IgM antibody between patients with schizophrenia and controls was not significant (P>0.005). There was no significant difference (P>0.005) in both serointensity of Tg IgG and DNA between patients with schizophrenia and controls. These findings have further demonstrated the strong association between the active Tg infection and schizophrenia.

Estudo clínico, laboratorial e epidemiológico da infecção por toxoplasma gondii em animais silvestres, bovinos, suínos e comunidades rurais da região de Nhecolândia, Pantanal, Brasil / Clinical, laboratory and epidemiological study of toxoplasma gondii infection in wild animals, cattle, pigs and rural communities in the region Nhecolândia, Pantanal, Brazil

A toxoplasmose, causada pelo Toxoplasma gondii é uma protozoose que acomete o homem e uma grande variedade de animais de sangue quente e aves. No Brasil, a prevalência pode variar de 20 por cento a 90 por cento dependendo da área estudada, clima, condição socioeconômica e cultural. A infecção se dá através da ingestão de oocistos, que podem ser encontrados no solo, água e alimentos ou através da manipulação e ingestão de carne crua ou mal cozida, além da infecção congênita, apresentando importância em saúde pública. Este trabalho objetivou estudar a ocorrência da infecção por Toxoplasma gondii em animais silvestres, bovinos, suínos, ovinos e comunidade rural da região de Nhecolândia, no Pantanal do Mato Grosso do Sul, utilizando métodos sorológicos (Hemaglutinação Indireta - HAI, Reação de Imunofluorescência Indireta - RIFI, Técnica de aglutinação modificada - MAT) e moleculares (Reação em cadeia pela polimerase – PCR, PCR-RFLP). Foram feitas coletas de amostras de sangue de 73 indivíduos da comunidade rural, de 25 cães, 442 bovinos e 148 porco-monteiros. Observou-se que 47,95 por cento (35/73) das pessoas eram sororreagentes. Destas, apenas um indivíduo sororreagente (2,9 por cento) apresentou lesão ocular presumível da infecção pelo parasito. Nos animais, observou-se a ocorrência de anticorpos anti- T. gondii em 48 por cento dos cães, 30,55 por cento dos bovinos e 1,3 por cento nos porco-monteiros. Relatos de várias partes do mundo têm demonstrado a importância do ciclo silvestre na epidemiologia da infecção por Toxoplasma gondii. No entanto, apesar do papel conhecido de alguns felinos selvagens como hospedeiros definitivos para manutenção e transmissão do parasita para outros predadores carnívoros, pouco se sabe sobre a incidência de Toxoplasma gondii nestes animais...

Toxoplasmosis, caused by Toxoplasma gondii is a protozoan infection that affects humans and a wide variety of warm-blooded animals and birds. In Brazil, the prevalence varies from20 percent to 90 percent depending on the study area, climate, socioeconomic and cultural conditions. Infection occurs through ingestion of oocysts, which can be found in soil, food and water orthrough manipulation and ingestion of raw or undercooked meat, with public health significance. Collection of blood samples from 73 individuals from the rural community of 25 dogs, 442 cattle and 148 feral pig Nhecolândia the region were made. It was found that47.95 percent (35/73) were seropositive persons. Of these, only one sororreagente individual (2.9 percent) had presumed ocular injury from infection by the parasite. In animals, we observed theoccurrence of 48 percent of dogs and 30.55 percent in cattle and 1.3 percent in feral pig. Reports from several parts of the world have demonstrated the importance of the sylvatic cycle in the epidemiology of Toxoplasma gondii infections. However, despite the known role of some wild felids as definitive hosts for maintenance and transmission of the parasite to other carnivore predators, little is known about the incidence of Toxoplasma gondii in these animals. Therefore, one of the objective of this study was to detect exposure and occurrence of the T. gondii infection inwild carnivores and rodents of the Pantanal techniques...

Genotyping of Toxoplasma gondii from Rats (Rattus rattus) in Riyadh, Saudi Arabia

Toxoplasma 3 main clonal lineages are designated as type I, II, and III; however, atypical and mixed genotypes were also reported. This study was conducted for detection of Toxoplasma gondii genotypes in rats (Rattus rattus) in Riyadh region, Saudi Arabia. PCR test on T. gondii B1 gene was conducted on ELISA IgM positive samples for confirmation of the infection. However, genetic analysis of the SAG2 locus was performed to determine T. gondii genotypes using PCR-RFLP technique. PCR test on T. gondii B1gene showed that 22 (81.5%) out of the 27 ELISA IgM positive samples have T. gondii DNA. Genotypic analysis shows that, of the total 22 PCR positive samples, only 13 (59.1%) were of type II, 7 (31.8%) were of type III, and 2 (9.1%) were of an unknown genotype. It is obvious that the prevalence of both type II and III is high in rats. No reports have been available on T. gondii genotypes among rats in Riyadh region, and only little is known about its seroprevalence in rats. Future studies on T. gondii genotypes in rats using multi-locus markers is needed in Riyadh region, Saudi Arabia for better understanding of T. gondii pathogenesis and treatment in humans and animals.

Toxoplasmose experimental no modelo murino com fenótipo extremo para tolerância oral: caracterização celular e humoral / Experimental toxoplasmosis in murine models selected for extreme phenotypes of oral tolerance: celular and humoral characterization

O tecido linfóide das mucosas apresenta estruturas especializadas contendo células imunorreguladoras e mecanismos capazes de determinar indução de tolerância ou resposta inflamatória após a administração oral de um antígeno. Diversos agentes causadores de doenças utilizam a via oral como rota de infecção, dentre eles o Toxoplasma gondii. A infecção por T. gondii pode gerar diversas manifestações patológicas, como por exemplo, a doença inflamatória intestinal, que envolve diferentes tipos celulares, diversas citocinas, bem como a participação da microbiota intestinal. Para melhor compreender o envolvimento dos diversos tipos celulares e elementos humorais na toxoplasmose, elegemos como modelos experimentais as linhagens de camundongos geneticamente selecionados para tolerância oral: TR (resistentes à tolerância) e TS (susceptíveis à tolerância). Os camundongos TS se caracterizam por baixa resposta inflamatória e maior quantidade de células T regulatórias produtoras de IL-10. Em contraste, os camundongos TR produzem fortes reações inflamatórias. A caracterização da toxoplasmose experimental nos camundongos TR e TS infectados por gavagem com T. gondii foi feita por análise histológica e imunohistoquímica de cortes do intestino, fígado e baço, dosagem de citocinas e quimiocinas, além da caracterização da microbiota intestinalNossos resultados confirmam o estabelecimento de uma resposta inflamatória intestinal aguda nos camundongos TR, que ocorre durante os 14 dias iniciais da infecção. As lesões apresentam macrófagos e, em menor número, células dendríticas. Além disso, o perfil de citocinas corrobora essas observações, com alta produção de citocinas pró-inflamatórias, tanto sistêmica quanto localizada. Os camundongos TS, por outro lado, apresentaram um perfil característico voltado para a regulação, inibindo a infecção intestinal e produção de citocinas inflamatórias, ao mesmo tempo em que favorece um aumento da quantidade de parasitos nos órgãos...

The mucosal lymphoid tissue has specialized structures containingimmunoregulatory cells and mechanisms to determine the induction of tolerance or inflammatory response following oral administration of an antigen. Several agentscausative of diseases uses the oral route as a form of infection, includingToxoplasma gondii. T. gondii infection can lead to various pathologicalmanifestations, such as inflammatory bowel disease, involving various cell types, several cytokines, as well as the involvement of the intestinal microbiota. To better understand the involvement of different cell types and humoral elements in the toxoplasmosis, we choose as experimental models two lineages of mice genetically selected for oral tolerance: TR (tolerance resistant) and TS (tolerance susceptible). TS mice are characterized by low inflammatory response and increased amount of regulatory T cells producing IL-10. In contrast, the TR mice produce strong inflammatory reactions. Characterization of experimentaltoxoplasmosis in TR and TS mice infected by gavage with T. gondii was made by histological and immunohistochemical analysis of sections of the intestine, liver andspleen, dosage of cytokines and chemokines, in addition to characterization of intestinal microbiota. Our results confirm the setting of an acute inflammatory response in the intestines of TR mice, which occurs during the initial 14 days from infection. Lesions presented macrophages and to a lesser number, dendritic cells. Moreover, thecytokine profile supports these observations, with a high production of proinflammatory cytokines, either systemic or localized...

Isolation and genotyping of Toxoplasma gondii from pregnant dairy cows (Bos taurus) slaughtered / Isolamento e genotipagem de Toxoplasma gondii em vacas de leite (Bos taurus) prenhas abatidas

The current study aimed to evaluate serology, and isolate and genotype Toxoplasma gondii strains from pregnant dairy cows, slaughtered in an abattoir for human consumption, and their fetuses. Blood from 60 pregnant dairy cows and blood and tissue samples (brain, lung, heart, and liver) from their fetuses were collected and analyzed in a mouse bioassay. Antibodies against T. gondii were observed in 48.3% of cows and 3.7% of fetuses (IFAT, titers ≥ 50 for cows and 25 for fetuses were considered positive). Fourteen fetuses (23.3%) and six cows (10.0%) were identified as positive in the bioassay. T. gondii was isolated from a blood sample of a cow older than 4 years old in the 6th month of pregnancy, and from a blood sample of a fetus in the 6th month of gestation. These isolates were identified by polymerase chain reaction (PCR) as being of T. gondii and both strains showed type II alleles for all PCR-restriction fragment length polymorphism (PCR-RFLP) markers tested. T. gondii type II strain from cattle was isolated for the first time in Brazil. The current study also showed that transplacental transmission of T. gondii naturally occurs in dairy cows (23.3%) from Southern Brazil.

O presente estudo teve como objetivo avaliar a ocorrência de anticorpos, isolar e genotipar Toxoplasma gondii de vacas prenhas abatidas em um matadouro para consumo humano e de seus respectivos fetos. Sangue de 60 vacas gestantes e amostras de sangue e tecidos (cérebro, pulmão, coração e fígado) de seus fetos foram coletados e utilizados para bioensaio em camundongos. Anticorpos contra T. gondii foram observados em 48,3% das vacas e em 3,7% dos fetos (foram considerados positivos títulos ≥50 para as vacas e ≥25 para os fetos). Quatorze fetos (23,3%) e seis vacas (10,0%) apresentaram-se positivas para T. gondii ao bioensaio. T. gondii foi isolado de amostra de sangue de uma vaca com mais de quatro anos no 6º mês de gestação e de amostra de sangue de um feto no 6º mês de gestação. Por PCR esses isolados foram identificados como sendo de T. gondii e ambas as cepas apresentaram o alelo tipo II em todos os marcadores de PCR-RFLP testados. Esta é a primeira identificação de genótipo tipo II de T. gondii em bovinos do Brasil. Além disso, este estudo mostrou que a transmissão transplacentária de T. gondii ocorre naturalmente em bovinos de leite (23,3%).

Toxoplasmose congênita em crianças sul-americanas: [revisão] / Congenital toxoplasmosis in south American children: [review]

Aims: To review the present knowledge about congenital toxoplasmosis in South America and to advance some hypothesis for future research. Source of data: Medline and Scielo database search for papers reporting clinical characteristics of cohorts of children in South America and comparative studies between South America and other continents. Summary of findings: Systematic analysis of primary data obtained during screening programs showed that the risk of ocular lesions in congenital toxoplasmosis was much higher in the South American cohorts (47%; 18/38) than in Europe (14%, 79/550). The crude risk of intracranial lesions was much higher in the cohorts from South America (53%, 20/38) than those from Europe (9%, 49/550). In a Colombian cohort it was found 11% of mortality. Additionally, a comparative prospective cohort of congenitally infected children from Brazil and Europe found that in Brazilian children eye lesions were larger, more numerous and more likely to affect the area of the retina responsible for central vision that their counterpart in Europe. The presence of Toxoplasma strains genetically different to those found in North America and Europe could explain the higher severity of congenital toxoplasmosis in South America. Conclusions: Congenital toxoplasmosis in South America seems to be more frequent and infected children are more symptomatic than in Europe and in North America. Research for new drugs and candidate vaccines are a priority to improve indicators of health in children of South America.

Objetivos: revisar o conhecimento atual sobre toxoplasmose congênita na América do Sul e traçar algumas hipóteses para futura pesquisa. Fonte de dados: busca nas bases de dados Pubmed e Scielo por artigos sobre características clínicas de coortes de crianças com toxoplasmose congênita na América do Sul e estudos comparativos entre América do Sul e outros continentes. Síntese dos dados: uma análise sistemática de dados primários obtidos durante programas de triagem mostrou que o risco de lesões oculares foi muito maior na coorte de crianças da América do Sul (47%, 18/38) do que nas européias (14%, 79/550). O risco bruto de lesões intracranianas foi muito maior na coortes da América do Sul (53%, 20/38) do que nas da Europa (9%, 49/550). Em uma coorte colombiana constatou-se 11% de mortalidade. Adicionalmente, uma coorte prospectiva, que comparou crianças com toxoplasmose congênita do Brasil e da Europa, mostrou que nas crianças brasileiras as lesões oculares foram maiores, mais numerosas e com maior probabilidade de atingir o polo posterior da retina do que nas européias. A presença de cepas de Toxoplasma gondii diferentes das da Europa e dos Estados Unidos pode explicar a maior gravidade da toxoplasmose congênita na América do Sul. Conclusões: a toxoplasmosis congênita na América do Sul parece ser mais frequente e as crianças infectadas são mais sintomáticas do que na Europa e na América do Norte. A pesquisa sobre novas drogas e vacinas deve ser prioritária, para melhorar os indicadores de saúde nas crianças da América do Sul.

Experimental reinfection of BALB/c mice with different recombinant type I/III strains of Toxoplasma gondii: involvement of IFN-³ and IL-10

To assess reinfection of BALB/c mice with different Toxoplasma gondii strains, the animals were prime infected with the non-virulent D8 strain and challenged with virulent recombinant strains. Thirty days after challenge, brain cysts were obtained from surviving BALB/c mice and inoculated in Swiss mice to obtain tachyzoites for DNA extraction and PCR-RFLP analysis to distinguish the different T. gondii strains present in possible co-infections. Anti-Toxoplasma immune responses were evaluated in D8-primed BALB/c mice by detecting IFN-³ and IL-10 produced by T cells and measuring immunoglobulin levels in serum samples. PCR-RFLP demonstrated that BALB/c mice were reinfected with the EGS strain at 45 days post prime infection (dpi) and with the EGS and CH3 strains at 180 dpi. High levels of IFN-³ were detected after D8 infection, with no significant difference between 45 and 180-day intervals. However, higher IL-10 levels and higher plasmatic IgG1 and IgA were detected from samples obtained 180 days after infection. BALB/c mice were susceptible to reinfection with different recombinant T. gondii strains and this susceptibility correlated with enhancement of IL-10 production.

Genotype of Toxoplasma gondii from Blood of Stray Cats in Gyeonggi-do, Korea

Genotyping of Toxoplasma gondii has been performed in 23 PCR positive blood samples from stray cats in Korea. We used 2 separate PCR-restriction fragment length polymorphism (RFLP) patterns of SAG2 gene, amplifying the 5'and 3'ends of the locus. The results revealed that all samples belonged to the type I clonal lineage. Although T. gondii organisms were not isolated from the samples, the results of the present study represent that stray cats with T. gondii infection should be seriously concerned in our environment. Adequate and continuous control programs of stray cats are needed to reduce the risk of transmission of T. gondii as a zoonotic infection threatening the public health.

Genotyping of a Korean isolate of Toxoplasma gondii by multilocus PCR-RFLP and microsatellite analysis

Although the Korean isolate KI-1 of Toxoplasma gondii has been considered to be a virulent type I lineage because of its virulent clinical manifestations, its genotype is unclear. In the present study, genotyping of the KI-1 was performed by multilocus PCR-RFLP and microsatellite sequencing. For 9 genetic markers (c22-8, c29-2, L358, PK1, SAG2, SAG3, GRA6, BTUB, and Apico), the KI-1 and RH strains exhibited typical PCR-RFLP patterns identical to the type I strains. DNA sequencing of tandem repeats in 5 microsatellite markers (B17, B18, TUB2, W35, and TgM-A) of the KI-1 also revealed patterns characteristic of the type I. These results provide strong genetic evidence that KI-1 is a type I lineage of T. gondii.

alteration of liver function indices in rat infected with Toxoplasma gondii [RH strain]

Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii. The migration of the parasite in blood as well as some organs like liver may cause some changes in physiological and biochemical indices in infected individuals. These may change the level of some indices like urea, total bilirubin, total protein, and albumin. The present investigation was conducted to study alteration of some liver functional indices of rats which were experimentally infected with Toxoplasma gondii. 80 non-infected rats and 60 non-infected mice were selected. Rats were divided into two groups of 60 test and 20 control rats. The test group was infected intraperitoneally with 5xl0[4] tachyzoites. Every 3 days for 60 days, three rats from test group and one rat from control group were bled. Standard techniques were used for urea, bilirubin, total protein and albumin tests. In addition, the livers of infected rats were searched biologically for presence of the parasite using intrapritoneal injection in mice method. The results indicated that, Toxoplasma cyst was present in the liver of infected rats within 6 to 27 days post infection. The parasite disappeared in liver after 28 days of infection. Biochemical results indicated that, urea from 6[th] to 60[th] day, total bilirubin from 6[th] to 27[th] day, albumin and total protein from 6[th] to 12[th] day post infection were increased but decreased to normal values afterward. Generally, temporary alteration of some biochemical indices during experimental infection of rat with toxoplasmosis may occur. The alteration mainly is due to the parasite migration to various tissues of the animal and it shifts to the normal condition following cyst formation in brain or muscles