Isolation and RFLP genotyping of Toxoplasma gondii from the domestic dogs (Canis familiaris) from Grenada, West Indies revealed high genetic variability

Abstract

Stray dogs are considered as sentinels in the epidemiology of Toxoplasma gondii because they are carnivores and eat a variety of foods, including garbage. In the present study, tissues and sera of 249 stray dogs (Canis familiaris) from Grenada, West Indies were examined for T. gondii infection. Sera were examined for antibodies to T. gondii by the modified agglutination test (MAT); 89 (35.7%) of 249 were seropositive with titers of 25 in seven dogs, 50 in 22 dogs, 100 in 22 dogs, 200 or higher in 38 dogs. Hearts of 76 seropositive dogs were bioassayed in mice. Viable T. gondii was isolated from 12 dogs; these isolates were designated TgDogGr1 to TgDogGr12. These isolates were further propagated in cell culture and DNA isolated from cell culture derived tachyzoites of 12 isolates was genotyped using 10 PCR-restriction fragment length polymorphism markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). The results revealed six genotypes, including ToxoDB PCR-RFLP #1, #2, #3, #7, #13 and #224, with 1, 6, 1, 2, 1 and 1 isolates, respectively. The result supports previous findings that T. gondii population genetics is highly diverse in Grenada.

Keywords: Dog (Canis familiaris); Genetic types; Grenada; Isolation; Seroprevalence; Toxoplasma gondii.