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Genetic divergence of Toxoplasma gondii strains associated with ocular toxoplasmosis, Brazil.

ABSTRACT: Previous studies have shown a high prevalence of toxoplasmosis and the frequent occurrence of ocular disease in Brazil. To identify the genotypes of parasite strains associated with ocular disease, we compared 25 clinical and animal isolates of Toxoplasma gondii from Brazil to previously characterized clonal lineages from North America and Europe. Multilocus nested polymerase chain reaction analysis was combined with direct sequencing of a polymorphic intron to classify strains by phylogenetic methods. The genotypes of T. gondii strains isolated from Brazil were highly divergent when compared to the previously described clonal lineages. Several new predominant genotypes were identified from different regions of Brazil, including 2 small outbreaks attributable to foodborne or waterborne infection. These findings show that the genetic makeup of T. gondii is more complex than previously recognized and suggest that unique or divergent genotypes may contribute to different clinical outcomes of toxoplasmosis in different localities.

SUBMITTER: Khan A

PROVIDER: S-EPMC3373049 | biostudies-literature | 2006 Jun

REPOSITORIES: biostudies-literature

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Genetic divergence of Toxoplasma gondii strains associated with ocular toxoplasmosis, Brazil.

Khan Asis A Jordan Catherine C Muccioli Cristina C Vallochi Adriana L AL Rizzo Luiz V LV Belfort Rubens R Vitor Ricardo W A RW Silveira Claudio C Sibley L David LD

Emerging infectious diseases 20060601 6

Previous studies have shown a high prevalence of toxoplasmosis and the frequent occurrence of ocular disease in Brazil. To identify the genotypes of parasite strains associated with ocular disease, we compared 25 clinical and animal isolates of Toxoplasma gondii from Brazil to previously characterized clonal lineages from North America and Europe. Multilocus nested polymerase chain reaction analysis was combined with direct sequencing of a polymorphic intron to classify strains by phylogenetic m ...[more]

PMID: 16707050

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Project description:BackgroundPrevious Toxoplasma gondii studies revealed that mutations in the dhps (dihydropteroate synthase) gene are associated with resistance to sulfonamides. Although Brazilian strains are genotypically different, very limited data are available regarding the susceptibility of strains obtained from human to sulfonamides. The aim of this study was to evaluate the efficacy of sulfadiazine (SDZ) against Brazilian isolates of T. gondii and verify whether isolates present polymorphisms in the dhps gene. We also investigated whether the virulence-phenotype and/or genotype were associated with the profile of susceptibility to SDZ.MethodsFive T. gondii isolates obtained from newborns with congenital toxoplasmosis were used to verify susceptibility. Mice were infected with 104 tachyzoites and orally treated with different doses of SDZ. The mortality curve was evaluated by the Log-rank test. The presence of polymorphisms in the dhps gene was verified using sequencing. A descriptive analysis for 11 Brazilian isolates was used to assess the association between susceptibility, genotype, and virulence-phenotype.ResultsStatistical analysis showed that TgCTBr03, 07, 08, and 16 isolates were susceptible to SDZ, whereas TgCTBr11 isolate presented a profile of resistance to SDZ. Nineteen polymorphisms were identified in dhps exons. Seven polymorphisms corresponded to non-synonymous mutations, with four being new mutations, described for the first time in this study. No association was found between the profile of susceptibility and the virulence-phenotype or genotype of the parasite.ConclusionsThere is a high variability in the susceptibilities of Brazilian T. gondii strains to SDZ, with evidence of drug resistance. Despite the large number of polymorphisms identified, the profile of susceptibility to SDZ was not associated with any of the dhps variants identified in this study. Other genetic factors, not yet determined, may be associated with the resistance to SDZ; thus, further studies are needed as a basis for a more adequate toxoplasmosis treatment.

Project description:Human toxoplasmosis, a protozoonosis caused by Toxoplasma gondii, has been described as a worldwide foodborne disease with important public health impact. Despite infection has reportedly varied due to differences in alimentary, cultural and hygienic habits and geographic region, social vulnerability influence on toxoplasmosis distribution remains to be fully established. Accordingly, the present study has aimed to assess T. gondii seroprevalence and factors associated to social vulnerability for infection in households of Ivaiporã, southern Brazil, with 33.6% population making half minimum wage or less, ranked 1,055th in population (31,816 habitants), 1,406th in per capita income (U$ 211.80 per month) and 1,021st in HDI (0.764) out of 5,570 Brazilian cities. Serum samples and epidemiological questionnaires were obtained from citizen volunteers with official City Secretary of Health assistance in 2015 and 2016. In overall, serosurvey has revealed 526/715 (73.57%) positive samples for anti-T. gondii antibodies by Indirect Fluorescent Antibody Test. Logistic regression has shown a significant increase associated to adults (p = 0.021) and elderly (p = 0.014) people, illiterates (p = 0.025), unemployment (p <0.001) and lack of household water tank (p = 0.039). On the other hand, sex (male or female), living area (urban or rural), yard hygiene, meat ingestion, sand or land contact, owning pets (dog, cat or both) were not significant variables of positivity for anti-T. gondii antibodies in the surveyed population. Although no significant spatial cluster was found, high intensity areas of seropositive individuals were located in the Kernel map where the suburban neighborhoods are located. In conclusion, socioeconomic vulnerability determinants may be associated to Toxoplasma gondii exposure. The increased risk due to illiteracy, adult or elderly age, unemployment and lack of household water tank were confirmed by multivariate analysis and the influence of low family income for seropositivity by the spatial analysis.

Project description:BackgroundToxoplasma gondii has a largely clonal population in North America and Europe, with types I, II and III clonal lineages accounting for the majority of strains isolated from patients. RH, a particular type I strain, is most frequently used to characterize Toxoplasma biology. However, compared to other type I strains, RH has unique characteristics such as faster growth, increased extracellular survival rate and inability to form orally infectious cysts. Thus, to identify candidate genes that could account for these parasite phenotypic differences, we determined genetic differences and differential parasite gene expression between RH and another type I strain, GT1. Moreover, as differences in host cell modulation could affect Toxoplasma replication in the host, we determined differentially modulated host processes among the type I strains through host transcriptional profiling.ResultsThrough whole genome sequencing, we identified 1,394 single nucleotide polymorphisms (SNPs) and insertions/deletions (indels) between RH and GT1. These SNPs/indels together with parasite gene expression differences between RH and GT1 were used to identify candidate genes that could account for type I phenotypic differences. A polymorphism in dense granule protein, GRA2, determined RH and GT1 differences in the evasion of the interferon gamma response. In addition, host transcriptional profiling identified that genes regulated by NF-ĸB, such as interleukin (IL)-12p40, were differentially modulated by the different type I strains. We subsequently showed that this difference in NF-ĸB activation was due to polymorphisms in GRA15. Furthermore, we observed that RH, but not other type I strains, recruited phosphorylated IĸBα (a component of the NF-ĸB complex) to the parasitophorous vacuole membrane and this recruitment of p- IĸBα was partially dependent on GRA2.ConclusionsWe identified candidate parasite genes that could be responsible for phenotypic variation among the type I strains through comparative genomics and transcriptomics. We also identified differentially modulated host pathways among the type I strains, and these can serve as a guideline for future studies in examining the phenotypic differences among type I strains.

Dataset Information

Genetic divergence of Toxoplasma gondii strains associated with ocular toxoplasmosis, Brazil.

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Genetic divergence of Toxoplasma gondii strains associated with ocular toxoplasmosis, Brazil.

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