Abstract

Toxoplasma gondii is an apicomplexan parasite that infects almost all warm-blooded animals. Little is known about how the parasite virulence in mice extrapolates to other relevant hosts. In the current study, in vitro phenotype and in vivo behavior in mice and sheep of a type II T. gondii isolate (TgShSp1) were compared with the reference type II T. gondii isolate (TgME49). The results of in vitro assays and the intraperitoneal inoculation of tachyzoites in mice indicated an enhanced virulence for the laboratory isolate, TgME49, compared to the recently obtained TgShSp1 isolate. TgShSp1 proliferated at a slower rate and had delayed lysis plaque formation compared to TgME49, but it formed more cyst-like structures in vitro. No mortality was observed in adult mice after infection with 1–105 tachyzoites intraperitoneally or with 25–2,000 oocysts orally of TgShSp1. In sheep orally challenged with oocysts, TgME49 infection resulted in sporadically higher rectal temperatures and higher parasite load in cotyledons from ewes that gave birth and brain tissues of the respective lambs, but no differences between these two isolates were found on fetal/lamb mortality or lesions and number of T. gondii-positive lambs. The congenital infection after challenge at mid-pregnancy with TgShSp1, measured as offspring mortality and vertical transmission, was different depending on the challenged host. In mice, mortality in 50% of the pups was observed when a dam was challenged with a high oocyst dose (500 TgShSp1 oocysts), whereas in sheep infected with the same dose of oocysts, mortality occurred in all fetuses. Likewise, mortality of 9 and 27% of the pups was observed in mice after infection with 100 and 25 TgShSp1 oocysts, respectively, while in sheep, infection with 50 and 10 TgShSp1 oocysts triggered mortality in 68 and 66% of the fetuses/lambs. Differences in vertical transmission in the surviving offspring were only found with the lower oocyst doses (100% after infection with 10 TgShSp1 oocysts in sheep and only 37% in mice after infection with 25 TgShSp1 oocysts). In conclusion, virulence in mice of T. gondii type II isolates may not be a good indicator to predict the outcome of infection in pregnant sheep.

Highlights

  • Toxoplasma gondii is an apicomplexan parasite capable of infecting almost all warm-blooded animals and causing potentially fatal disease in humans and some relevant domestic species, such as small ruminants (Dubey, 2010)

  • On day 40 post-inoculation, one mouse inoculated with the brain homogenate of the sheep abortion was T. gondii PCR-positive in the brain

  • TgShSp1 cultured under regular conditions demonstrated spontaneous conversion to bradyzoite with a statistically higher number of Dolichos biflorus lectin (DBL)-positive cysts (14%) compared to TgME49 (2%) (P < 0.0001)

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Summary

Introduction

Toxoplasma gondii is an apicomplexan parasite capable of infecting almost all warm-blooded animals and causing potentially fatal disease in humans and some relevant domestic species, such as small ruminants (Dubey, 2010). Type I isolates are highly virulent in mice, whereas types II and III show a dose-dependent mortality (Saeij et al, 2006). Laboratory isolates from types I, II, and III are used in T. gondii research and have been maintained in successive passages in cell culture and mice. Enhanced virulence throughout successive passages in cell culture and mice for T. gondii type I isolates (e.g., RH isolate) has been widely reported (Villard et al, 1997; Mavin et al, 2004; Khan et al, 2009), but little is known about the influence of continuous passages in type II isolates. There is little information on how T. gondii virulence in mice compares to virulence in other species, in those experiencing clinical toxoplasmosis, such as sheep and humans

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