Sarcocystidae in wild birds of southeastern Brazil.

This study aimed to identify members of the Sarcocystidae family in naturally infected wild birds at a rescue center in the state of Minas Gerais, southeastern Brazil. The heart and brain of 44 wild birds were evaluated by bioassay in mice to detect T. gondii, and extracted DNA was used for nested PCR of the 18S ribosomal DNA gene to detect members of the Sarcocystidae family. The positive samples were sequenced, assembled, edited and compared with sequences deposited in GenBank. Toxoplasma gondii was isolated from six (13.6%) out of 44 birds. Toxoplasma gondii DNA was identified in 10/44 (22.7%) of the birds. The amplified sequences exhibited 100% similarity with the DNA of the ME49 strain of T. gondii. Sarcocystis DNA (99% similarity) was identified in 5/44 (11.4%) of the birds. T. gondii and Sarcocystis spp. are common in wild birds in Minas Gerais, Brazil.

Toxoplasma gondii: Cytokine responses in mice reinfected with atypical strains.

This study aimed to elucidate the cellular immune response against Toxoplasma gondii in chronically infected mice reinfected with different strains of the parasite to elucidate the immunological basis for chronicity or virulence and to uncover the involvement of genes that encode virulence proteins and modulate the immune response. BALB/c mice were infected by oral gavage with non-virulent D8 strain and challenged 45 days post-infection with virulent EGS or CH3 strains. Cytokine measurement was performed 2 days post-challenge in cell extracts of the small intestine and 2, 7, and 14 days post-challenge in serum. Virulence gene allele type of these strains was analyzed. Challenged mice survived by avoiding exacerbated inflammation and inhibiting the overproduction of cytokines. Local and systemic cytokine response in challenged mice was similar to chronic controls and quite distinct in mice acutely infected with the EGS or CH3 strains. Allelic combinations of the virulence genes ROP5/ROP18 was predictive of virulence in mice when tested in these T. gondii strains. Other allelic combinations of rhoptries and dense granules genes showed discrepancies.

Tissue dissemination and humoral response after experimental reinfection with atypical Toxoplasma gondii strains obtained from congenital human toxoplasmosis in Brazil.

The paradigm that Toxoplasma gondii infection generates sterilizing protective immunity was broken by case studies in which reinfections were observed in immunocompetent pregnant women in the chronic phase of toxoplasmosis. Since then, several murine models have suggested that immunoprotection against a previous T. gondii infection may be violated after reinfection with strains of different genotypes. This study aimed to evaluate the dissemination of the parasite after reinfection with the virulent TgCTBr9 and EGS strains in BALB/c mice chronically infected with the avirulent TgCTBr5 strain. Three mice were euthanized at 2, 4, 8, 12, 24 and 48 h post challenge (p.c.) and at 7, 14 and 30 days p.c. Intestines, mesenteric lymph nodes, lungs and brains were collected for PCR-RFLP. Blood samples were collected to measure total IgG, IgG1 and IgG2a by ELISA. The reinfected animals survived and presented reduced morbidity after challenge with the virulent strains. Mice challenged with the TgCTBr9 strain showed a slight increase in anti-T. gondii IgG1. The spread of the TgCTBr5 strain was observed to occur earlier than the dissemination of the virulent TgCTBr9 or EGS strains. The TgCTBr9 strain was observed in the mesenteric lymph node at 7 days post challenge (d.p.c.); in the intestine and lungs at 14 d.p.c.; and in the brain at 30 d.p.c. EGS strain was demonstrated in the mesenteric lymph node and lung at 7 d.p.c and in the intestine and brain at a later time point. The immune response promoted by the primary infection with the avirulent strain (TgCTBr5) protected the animals from death after challenge with the virulent strains (TgCTBr9 or EGS).

Surto de toxoplasmose no município de Gouveia, Minas Gerais / Outbreak of toxoplamosis in Goveia, Minas Gerais

Objetivo: investigar surto de toxoplasmose no município de Gouveia-MG, no período entre fevereiro e maio de 2015. Método: foram realizados estudos descritivo e caso-controle, com análise multivariada (regressão logística) e nível de significância de 5%. Definiu-se como caso o indivíduo com toxoplasmose aguda, confirmada laboratorialmente (IgM e/ou IgG reagentes e baixa avidez de IgG), e controle o indivíduo susceptível (IgM e IgG não reagentes). Resultado: o estudo contemplou 50 casos e 68 controles, sendo a maioria dos casos do sexo masculino (43 pacientes, 86%). Os casos tiveram 10 vezes mais chance de tomar água procedente do Manancial (IC95%: 2,8-37,4; p=0,01), 5,1 vezes mais chance de ingerir linguiça (IC95%: 1,2-20,4; p=0,02) e 4,5 vezes mais chance de ser do sexo masculino (IC95%: 1,2-16,7; p=0,02), quando comparados aos controles. Conclusão: foi confirmado um surto de toxoplasmose aguda, em Gouveia - MG, de transmissão hídrica e/ou alimentar. A maior ocorrência no sexo masculino pode estar relacionada a hábitos de risco, envolvidos na transmissão da doença. Foram propostas medidas que resultaram na diminuição e no controle dos casos da doença.

Objective: to investigate the outbreak of toxoplasmosis in the municipality of Gouveia-MG, between February and May 2015. Methods: Descriptive and case control study, with multivariate analysis (logistic regression) and level of significance of 5%. For the purpose of the study, a case was defined on an individual with laboratory which confirmed acute toxoplasmosis (IgM and/or IgG with low IgG avidity) and a control was a susceptible individual (nonreactive IgM and IgG). Results: The study comprised 50 cases and 68 controls, with most cases being male (43 patients, 86%). When compared to controls, cases had 10 times greater odds of having drunk water from Stream A (IC95%: 2,8-37,4; p=0,01), 5,1 times greater odds of having ingested sausage (IC95%: 1,2-20,4; p=0,02) and 4.5 times greater odds of being of male sex (IC95%: 1,2-16,7; p=0,02). Conclusion: A water/foodborne outbreak of acute toxoplasmosis was confirmed in Gouveia-MG, affecting predominantly men. This may be associated with habits regarding disease transmission. Measures that resulted in the reduction and control of cases of this disease have been proposed.

Toxoplasma gondii: isolation, biological and molecular characterisation of samples from free-range Gallus gallus domesticus from countryside Southeast Brazil / Toxoplasma gondii: isolamento, caracterização biológica e molecular de amostras de Gallus gallus domesticus no interior do Sudeste brasileiro

Abstract Toxoplasma gondii presents a high prevalence worldwide, infecting several animals. Felines are considered the definitive hosts and among the intermediate hosts we highlight mammals and birds. The man can become infected by ingesting tissue cysts present in birds and mammals. Biological and molecular aspects of T. gondii allows a better understanding of the epidemiology of toxoplasmosis. This work is a serologic screening of 58 chickens grown (Gallus gallus domesticus) for human consumption in Espírito Santo State, by means of indirect haemagglutination assay (IHA). Thirteen chickens tested positive for anti-T. gondii antibodies. The heart and brain of five positive chickens were harvested, treated with pepsin and inoculated separately, in two Swiss mice, intraperitoneally. Tachyzoites were observed in the peritoneum of all the animals, between seven and 10 days after the inoculum. Ten isolates were obtained and biologically characterised in BALB/c mice inoculated with 101 to 104 tachyzoites. All isolates were classified as virulent or intermediately virulent. Isolates were genotyped by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, revealing three different genotypes. None of the isolates exhibited the clonal type I, II or III genotype. No genotypic differences were observed between the isolates from the brain or heart from the same bird.

Resumo Toxoplasma gondii apresenta alta prevalência mundial, capaz de infectar diversos animais. Felinos são considerados os hospedeiros definitivos e entre os hospedeiros intermediários destacamos os mamíferos e as aves. O homem pode se infectar ingerindo cistos teciduais presentes na carne das aves e mamíferos. O conhecimento dos aspectos biológicos e moleculares do parasito possibilitam melhor entendimento da epidemiologia da toxoplasmose. Neste trabalho foi realizada triagem sorológica por hemaglutinação indireta (HI) em 58 galinhas caipiras (Gallus gallus domesticus) utilizadas para consumo humano, provenientes do estado do Espírito Santo, Brasil. Treze galinhas apresentaram sorologia positiva para T. gondii. O coração e o cérebro de cinco galinhas positivas foram colhidos, tratados com pepsina e inoculados separadamente, em dois camundongos Swiss, por via intraperitoneal. Observou-se taquizoítos no peritônio de todos os camundongos, entre sete e 10 dias após o inóculo. Foram obtidos 10 novos isolados de T. gondii os quais foram estudados em camundongos BALB/C inoculados com 101 a 104 taquizoítos por animal. Todos os isolados foram considerados virulentos ou de virulência intermediária. A caracterização molecular dos isolados, realizada por PCR-RFLP, demonstrou a ocorrência de três genótipos distintos. Nenhum isolado apresentou genótipo clonal ou linhagem clonal do Brasil. Não foi observada diferença molecular (PCR-RFLP) entre os isolados obtidos a partir do cérebro ou do coração da mesma ave. Dois isolados já haviam sido relatados na literatura como causadores de doenças em humanos.

Toxoplasma gondii: isolation, biological and molecular characterisation of samples from free-range Gallus gallus domesticus from countryside Southeast Brazil.

Toxoplasma gondii presents a high prevalence worldwide, infecting several animals. Felines are considered the definitive hosts and among the intermediate hosts we highlight mammals and birds. The man can become infected by ingesting tissue cysts present in birds and mammals. Biological and molecular aspects of T. gondii allows a better understanding of the epidemiology of toxoplasmosis. This work is a serologic screening of 58 chickens grown (Gallus gallus domesticus) for human consumption in Espírito Santo State, by means of indirect haemagglutination assay (IHA). Thirteen chickens tested positive for anti-T. gondii antibodies. The heart and brain of five positive chickens were harvested, treated with pepsin and inoculated separately, in two Swiss mice, intraperitoneally. Tachyzoites were observed in the peritoneum of all the animals, between seven and 10 days after the inoculum. Ten isolates were obtained and biologically characterised in BALB/c mice inoculated with 101 to 104 tachyzoites. All isolates were classified as virulent or intermediately virulent. Isolates were genotyped by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, revealing three different genotypes. None of the isolates exhibited the clonal type I, II or III genotype. No genotypic differences were observed between the isolates from the brain or heart from the same bird.

Kinetics of parasite distribution after reinfection with genetically distinct strains of Toxoplasma gondii.

Recent data shows that prior infection by Toxoplasma gondii does not protect the host from subsequent reinfection even after the development of immunological memory. Although animal models for T. gondii reinfection were proposed after cases of natural human reinfection were described, little is known about the events that occur immediately after challenge. To further understand these events, BALB/c mice were chronically infected with D8 non-virulent strain (genotype ToxoDB#8 BrIII) and challenged with two different virulent strains: EGS (genotype ToxoDB #229) or CH3 strain (genotype ToxoDB #19). Primary infection protected animals from lethal challenge and morbidity was reduced. Reinfection was confirmed by PCR-RFLP, showing differences in the way the parasites spread in challenged animals. Parasites reached the lungs during early infection and a parasitism delay in the intestine was observed in D8+CH3 group. Parasites from challenge strains were not detected in the brain of D8+CH3 and in the intestine and brain of D8+EGS group. Previous infection with D8 strain of T. gondii protected against lethal challenges, but it did not prevent parasite spread to some organs.