Characterization of Toxoplasma gondii from Raccoons (Procyon lotor), Coyotes (Canis latrans), and Striped Skunks (Mephitis mephitis) in Wisconsin Identified Several Atypical Genotypes

Abstract

During 2005-2006, sera and tissues from raccoons (Procyon lotor), coyotes (Canis latrans), and skunks (Mephitis mephitis) from the state of Wisconsin were tested for Toxoplasma gondii infection. Antibodies to T. gondii were found in 32 of 54 (59.2%) raccoons, 18 of 35 (51.4%) coyotes, and 5 of 7 (71.4%) skunks using the modified agglutination test and a cut-off titer of 1:20. Pooled tissues (brains, hearts, and tongues) from 30 raccoons, 15 coyotes, and 1 skunk were bioassayed for T. gondii infection in mice or cats. Viable T. gondii was isolated from 5 of 30 (16.7%) raccoons, 6 of 15 (40.0%) coyotes, and the skunk. Genetic characterization of the 12 parasite isolates by multilocus PCR-RFLP markers revealed 6 different genotypes including 5 atypical and I archetypal II lineages. The results indicate the prevalence of T. gondii in wildlife mammals is high and that these animals may serve as an important reservoir for transmission of T. gondii.

Genotyping Toxoplasma gondii from wildlife in Pennsylvania and identification of natural recombinants virulent to mice

“…Laboratory-reared coyotes fed T. gondii oocysts or tissue cysts became infected but remained asymptomatic (Dubey, 1982). In the present study, T. gondii antibodies were found in 34.5% of 266 coyotes, which is lower than 51.4% of 35 coyotes from Wisconsin (Dubey et al, 2007), 59% of 222 coyotes collected in Kentucky, Ohio, and Indiana (Dubey et al, 1999), and 62% of 52 coyotes from Texas (Lindsay et al, 1996). In the present study viable T. gondii was isolated from 10 (32.3%) of 53 seropositive coyotes.…”

“…In the present study viable T. gondii was isolated from 10 (32.3%) of 53 seropositive coyotes. The majority of isolates that have been genotyped (18 in total) in the present or previous studies (Dubey et al, 2004a, 2007, 2011) comprise genotypes #1 and #3 are also known as the Type II, genotypes #4 and #5 are known as Type 12, both are dominant in wildlife in North America (Dubey et al, 2011). …”

Application of a new PCR-RFLP panel suggests a restricted population structure for Eimeria tenella in UK and Irish chickens

“…There are more than 170 peer reviewed manuscripts available in PubMed under the keywords ‘ Toxoplasma’ and ‘PCR-RFLP’ (accessed 12th July; 2016); including surveillance of multiple host species in many different regions (e.g. Su et al, 2006, Dubey et al, 2007a, Dubey et al, 2007b, Dubey et al, 2008, Velmurugan et al, 2009, Burrells et al, 2016). Development of PCR-RFLP for Eimeria ; starting with E. tenella ; will allow expansion of genetics studies with these parasites.…”

“…The approach is straightforward and applicable in many laboratories given provision of a thermal cycler and equipment for gel electrophoresis. Among apicomplexan parasites PCR-RFLP has been employed most notably to genotype T. gondii , including examples of parasites derived from multiple host species (Su et al, 2006, Dubey et al, 2008, Burrells et al, 2016) and regions (Dubey et al, 2007a, Dubey et al, 2007b, Velmurugan et al, 2009), and underpinning some of the most fundamental population genetics studies (Su et al, 2006). The availability of more accessible and cost-effective population genetics tools will provide opportunities for complimentary, and importantly comparable, analyses of E. tenella from other populations.…”

Reinfecção experimental de camundongos Balb/c por cepas geneticamente distintas de Toxoplasma gondii

“…As sequências de DNA alvo foram, primeiramente, amplificadas em dois multiplex PCR, utilizando primers externos para todos os marcadores (multiplex 1: SAG1, 5' 3' SAG2, SAG3, BTUB e GRA6 e multiplex 2: alt.SAG2, c22-8, c29-2, L358, PK1 e Apico), seguido de nested PCR para cada marcador, individualmente, utilizando primers internos, conforme descrito previamente (110,198,199). Os detalhes sobre os marcadores moleculares, primers (PCR e nested PCR) e enzimas de restrição que foram utilizadas estão presentes no Quadro 1…”