Toxoplasma gondii is a protozoon parasite that can cause severe clinical problems such as congenital toxoplasmosis. the distribution of T. gondii genotypes varies from one geographic area to another. So far, little is known about the parasite genotypes in tunisia, north Africa. the present study aimed isolating and genotyping T. gondii from the amniotic fluid (AF) and placenta of pregnant women in Monastir, Tunisia. Amniotic fluid and/or placenta from 80 women who acquired toxoplasma infection during pregnancy were tested by PCR and/or mouse bioassay. Genotyping of T. gondii isolates from these samples was performed with 15 microsatellite markers. Four viable T. gondii strains were isolated from either the AF or placenta of four women. Specifically, strains TUN001-MON1 and TUN002-MON2 were isolated from both the AF and placenta, TUN003-AHA from only the placenta, and TUN004-NEL from only the AF. The four viable strains were not virulent for mice. Genotyping revealed that the four strains were type II strains. This is the first report on isolation and genotyping of T. gondii from Af human samples in tunisia. further studies focused on T. gondii genotyping on a larger number of human cases and on animals in tunisia are needed to improve the knowledge and epidemiology of toxoplasmosis. Toxoplasma infection is one of the most prevalent parasitic disease, caused by the intracellular protozoa, Toxoplasma gondii. This protozoan affects all warm-blooded animals, including humans 1. Humans are infected by the ingestion of Toxoplasma oocysts in water, food or cat feces polluted soil, or toxoplasma cysts present in raw or undercooked meat 2. Congenital toxoplasmosis (CT) may happen following maternal primary infection during pregnancy. Risk of transmission increases with the pregnancy age, while severity of the disease for the fetus decreases. In fact, placenta barrier is more efficient at the first semester of gestation, allowing the passage of parasites in less than 10% of infected pregnant women. However, it becomes more permeable during pregnancy evolution, leading to parasite transmission around 30% and 60-70% of infected pregnant women in the second and third trimester respectively 3. Although, most congenitally infected newborns appear to be healthy at birth, they may develop symptoms until months, years, or even decades later in life. Hydrocephalus, intracranial calcifications, mental retardation, hepatosplenomegaly, and chorioretinitis are classical signs associated with the disease 3. Undiagnosed and untreated patients may expand irreversible lesions, particularly brain calcifications, hydrocephalus, and eye disease 4. Severity of CT may be associated to several factors including parasite genotype, host genetic variability and immune response 5-7. Previously, T. gondii complex was classified into three major lineages designated type I, II and III 8. However, recent studies using various molecular tools and analyzing genetic
The isolation and molecular typing of Toxoplasma gondii strains provide an essential basis for a better understanding of the parasite’s genetic diversity, determinants of its geographical distribution and associated risks to human health. In this study, we isolated and genetically characterized T. gondii strains from domestic animals in Southern and coastal area of Tunisia. Blood, hearts and/or brains were collected from 766 domestic animals (630 sheep and 136 free-range chickens). Strain isolation from these samples was performed using mouse bioassay and genotyping was carried out with a multiplex PCR technique using 15 microsatellite markers. Thirty viable strains of T. gondii were successfully isolated from tissues of sheep (19/142) and chickens (11/33). In addition, 3 strains could be successfully genotyped from animal tissues for which mouse bioassay was unsuccessful. A large predominance of type II strains (n = 29) was found in the sampled regions, followed by type III (n = 3) and, for the first time in Tunisia, a single isolate of Africa 4 lineage from a sheep. Analyses of population genetics showed the presence of a divergent population of type II lineage in Tunisia, supporting limited recent migrations of strains between Tunisia and other countries of the world.
Toxoplasma gondii is a protozoan parasite that can be transmitted to humans through a variety of routes including blood transfusion. This study aimed to investigate the seroprevalence of T. gondii infection and associated risk factors in healthy blood donors in Tunisia. A total of 800 healthy blood donors from two blood centers in south and coastal Tunisia were analyzed for anti-T. gondii IgG and IgM antibodies by indirect immunofluorescence assay (IFA) and enzyme-linked immunoassays (ELISA), respectively. Structured questionnaires were used to gather information on risk factors for T. gondii infection during collection. The overall seroprevalence was 44.4% of which 352 (44%) and 3 (0.4%) were positive for IgG and both IgG and IgM anti-T. gondii antibodies, respectively. Multivariate analysis showed that T. gondii seropositivity was significantly associated with the birth place (adjusted odds ratio [OR] = 2.72; 95% confidence interval [CI]: 1.49–4.94) and the age of the donors (adjusted OR = 4.98; 95% CI: 1.50–16.58) which are independent risk factors. In addition, the variables of hand washing before eating (adjusted OR = 0.52; 95% CI: 0.37–0.74) and living in an urban environment (adjusted OR = 0.30; 95% CI: 0.13–0.71) are two protective factors. This study provided the first data on the seroprevalence and epidemiology of T. gondii infection in healthy blood donors in Tunisia.
The isolation and molecular typing of Toxoplasma gondii strains provides an essential basis for a better understanding of the distribution of genetic diversity and associated human health risks. In this context, we report, the molecular characterization of strains of T. gondii isolated from domestic animals infected with T. gondii in Southern and coastal area of Tunisia. This diversity is compared to that known for the Western European and Mediterranean sub-region.Blood, hearts and/or brains were collected from 766 domestic animals (630 sheep and 136 free-range chickens). Strain isolation from these samples were performed using mouse bioassay. The strains genotyping was carried out with a multiplex PCR technique using 15 microsatellite markers. Viable strains of T. gondii were isolated from 13.4% sheep and 33.3% chickens. Furthermore, the parasite was also detected in three DNA extracts from animal tissue digestates.This study showed a large predominance of type II strains (87.9%) from which two samples were type II variants for W35 locus. The other genotypes were three type III sheep isolates (9.1%) and, for the first time in Tunisia, an isolate of sheep origin of the Africa 4 genotype (3.0%). The comparison of microsatellite alleles of type II strains shows the recent migration of strains between Tunisia and other countries of the world.
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