Genotyping Toxoplasma gondii with the B1 Gene in Naturally Infected Sheep from an Endemic Region in the Pacific Coast of Mexico.

Toxoplasma gondii is a protozoan parasite with a broad ecological valence, which has been detected in a wide range of hosts and landscapes. Although the genus is considered monospecific, in recent years it has been demonstrated to exhibit more genetic variability than previously known. In Mexico, there are few genotyping studies, which suggest that classical, autochthonous, and atypical strains are circulating. The goal of this study was to describe T. gondii genetic diversity in naturally infected sheep from Colima, Mexico. This is a good site to study ecological aspects of this parasite since it is located between the Nearctic and Neotropical ecozones and it includes domestic and wild risks for transmission. We analyzed 305 tissue samples of semicaptive sheep from six coastal and central zones of Colima and border zones of Michoacán. We used an 803 bp amplicon of the B1 gene to genotype T. gondii and seroprevalence was determined by ELISA. Indexes for genetic diversity and genetic differentiation were calculated and compared with reference strains from North America (NA) and South America (SA). Twenty-three tissue samples were positive for the B1 gene by PCR, which were sequenced. Crude prevalence was 24.4%. The genetic analysis showed 16 variable sites along the 803 bp region that grouped all sequences into 13 haplotypes in the phylogenetic tree. Bayesian and haplotype network analysis showed nine new B1-types, of which three were frequent and six had unique alleles. Comparisons among sequence sets revealed that the Mexican population had lower differentiation than SA and an intermediate genetic variability between South America and North America. The B1 gene analysis showed new T. gondii haplotypes in naturally infected sheep; therefore, this marker could be initially used in molecular screening studies to identify potentially virulent genotypes of this parasite using natural host samples directly.

First isolation of Toxoplasma gondii from cats of Colima, Mexico: tissue distribution and genetic characterization.

Toxoplasma gondii is among the commonest zoonotic infectious agents worldwide. It infects many warm-blooded animals, including felines, the definitive hosts. This parasite is now classified in 15 haplogroups spread out around the world. Few reports reveal a predominance of genotypes I and III in Mexico, although recombinant and atypical variants have also been found in humans and animals. The aim of this study was to detect, isolate and genotype T. gondii from cats of Colima Mexico, and to analyze tissue distribution of the parasite. IgG specific antibodies were investigated in 48 serum samples from unwanted and stray cats by indirect ELISA. Isolation in mice and molecular characterization by PCR-RFLP and sequencing were attempted using pools of brain, heart, liver, lung, spleen and brachiocephalic muscle samples of seropositive cats. Fourteen animals (29.2%) were seropositive, the frequency ranged between 27.3 and 40% among the different localities. Ten seropositive animals were euthanized, eight of them were positive for the B1 gene by conventional PCR. More frequently infected tissues were the brachiocephalic muscle (75.0%) the brain (63.0%) and the spleen (63.0%). Genotype III was determined for the SAG3 locus of the parasite infecting an unwanted cat. Tachyzoites were isolated from the peritoneal cavity of two mice inoculated with the tissue pool of one kitten. Type I alleles were found in SAG1, SAG2, SAG3, BTUB, GRA6, c29-2 and PK1 loci, while c22-8 was type II, and L358 and Apico were type III. This genotype corresponds to ToxoDB genotype #28. This is the first T. gondii isolate genetically characterized in Colima, Mexico and is different to other isolations of the country.