No abstract

No abstract

Toxoplasma gondii is a widespread parasite of animals that causes zoonotic infections in humans. Previous studies have revealed a strongly clonal population structure in North America and Europe, while strains from South America are genetically separate and more diverse. However, the composition within North America has been questioned by recent descriptions of genetically more variable strains from this region. Here, we examined an expanded set of isolates using sequencedbased phylogenetic and population analyses to re-evaluate the population structure of T. gondii in North America. Our findings reveal that isolates previously defined by atypical restriction fragment length polymorphism patterns fall into two discrete groups. In one case, these new isolates represent variants of an existing lineage, from which they differ only by minor mutational drift. However, in the second case, it is evident that these isolates define a completely new lineage that is common in North America. Support for this new lineage was based on phylogeny, principle components analysis, STRUCTURE analyses, and statistical analysis of gene flow between groups. This new group, referred to as haplogroup 12, contains divergent genotypes previously referred to as A and X, isolated from sea otters. Consistent with this, group 12 was found primarily in wild animals, as well as occasionally in humans. This new lineage also has a highly clonal population structure. Analysis of the inheritance of multilocus genotypes revealed that different strains within group 12 are the products of a single recombination event between type 2 and a unique parental lineage. Collectively, the archetypal type 2 has been associated with clonal expansion of a small number of lineages in the North, as a consequence of separate but infrequent genetic crosses with several different parental lines.

No abstract

Toxoplasma infection is c o m m o n in m a n and animals, yet for 60 years the life cycle of Toxoplasma gondii remained unknown. R e c e n t l y a new form of Toxoplasma was found in the feces of cats that had eaten Toxoplasma-infected mice (for review of earlier work see [1]). This fecal form is biologically different from the known stages of Toxoplasma. While searching the feces of cats for a morphological equivalent of Toxoplasma, several candidate forms such as fungi, cysts of flagellates, and coccidian oocysts resembling those of Isospora fells, I. rivolta, and I. bigemina were found. Of these only oocysts resembling [. bigemina were constantly and q u a n t i t a t i v e l y associated with fecal Toxoplasma infectivity. W e will describe and characterize these oocysts and show b y a series of m u t u a l l y independent determinations that they should be regarded as oocysts of Toxoplasma got~dii. Some of these findings were briefly reported (2). DEFINITION O F TER3£STrophozoites refer to intracellular and free forms of Toxoplasma which are actively proliferating in the tissues of acutely infected animals (Fig. 1). Free trophozoites are quickly digested in solutions of pepsin at pH 1.3.Cyst refers to an accumulation of Toxoplasma (merozoites) characteristically occurring in the brain and muscle of chronically infected animals (Fig. 2). Cysts are surrounded by an elastic argyrophilic and periodic acid Schiff positive wall and contain much stored glycogen. The cyst wall is destroyed immediately on exposure to pepsin but the released merozoites survive in it for some time.

There is a lack of information about the seroepidemiology of T. gondii infection in the general population of Durango City, Mexico. Anti- Toxoplasma gondii IgG and IgM antibodies were sought in 974 inhabitants in Durango City, Mexico with the use of enzyme-linked immunoassays. in total, 59 (6.1%) of 974 participants (mean age 37 ± 16.1 yr) had IgG anti- T. gondii antibodies. Twenty (2.1%) of them also had IgM anti- T. gondii antibodies. IgG levels of 13-99, 100-150, and >150 International Units (IU)/ml were found in 14 (23.7%), 3 (5.1%), and 42 (71.2%) anti- T. gondii IgG-positive participants, respectively. Prevalence of infection increased with age (P < 0.05), and was significantly lower in participants born in Durango State than those born in other Mexican states (P < 0.01). Toxoplasma gondii infection was significantly associated with consumption of boar meat (adjusted odds ratio [OR]  =  3.02; 95% confidence interval [CI]: 1.49-6.13), and squirrel meat (adjusted OR  =  2.18; 95% CI: 1.17-4.09). in addition, infection was negatively associated with travel abroad (adjusted OR  =  0.42; 95% CI: 0.23-0.77), and salami consumption (adjusted OR  =  0.57; 95% CI: 0.32-0.99). This is the first report of seroprevalence and contributing factors for T. gondii infection in general population in Durango City, and of an association of the consumption of boar meat with T. gondii infection. This study provides a basis for the design of successful preventive measures against T. gondii infection.

To study the effect of high temperature on infectivity of Toxoplasma gondii tissue cysts, pork from infected pigs was mixed with infected mouse brains and homogenized thoroughly. Twenty-gram samples of infected homogenized meat were sealed in plastic pouches, pressed to a uniform thickness of 2 mm, and subjected to water-bath temperatures of 49, 52, 55, 58, 61, 64, and 67 C for 0.01, 3, 6, 9, 12, 24, 48, and 96 min. Treated samples were digested in HCl-pepsin solution and bioassayed in mice. Toxoplasma gondii tissue cysts remained viable at 52 C for 9.5 min but not for 9.5 min at 58 C; tissue cysts were generally rendered nonviable by heating to 61 C or higher temperature for 3.6 min. Tissue cysts survived once at 64 C for 3 min. These data demonstrate that T. gondii tissue cysts are less heat resistant than encysted Trichinella spiralis larvae.

a b s t r a c tLittle is known of the genetic diversity of Toxoplasma gondii circulating in wildlife. In the present study wild animals, from the USA were examined for T. gondii infection. Tissues of naturally exposed animals were bioassayed in mice for isolation of viable parasites. Viable T. gondii was isolated from 31 animals including, to our knowledge for the first time, from a bald eagle (Haliaeetus leucocephalus), five gray wolves (Canis lupus), a woodrat (Neotoma micropus), and five Arctic foxes (Alopex lagopus). Additionally, 66 T. gondii isolates obtained previously, but not genetically characterised, were revived in mice. Toxoplasma gondii DNA isolated from these 97 samples (31 + 66) was characterised using 11 PCR-restriction fragment length polymorphism (RFLP) markers (SAG1, 5 0 -and 3 0 -SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). A total of 95 isolates were successfully genotyped. In addition to clonal Types II, and III, 12 different genotypes were found. These genotype data were combined with 74 T. gondii isolates previously characterised from wildlife from North America and a composite data set of 169 isolates comprised 22 genotypes, including clonal Types II, III and 20 atypical genotypes. Phylogenetic network analysis showed limited diversity with dominance of a recently designated fourth clonal type (Type 12) in North America, followed by the Type II and III lineages. These three major lineages together accounted for 85% of strains in North America. The Type 12 lineage includes previously identified Type A and X strains from sea otters. This study revealed that the Type 12 lineage accounts for 46.7% (79/169) of isolates and is dominant in wildlife of North America. No clonal Type I strain was identified among these wildlife isolates. These results suggest that T. gondii strains in wildlife from North America have limited diversity, with the occurrence of only a few major clonal types. Ó