Suchergebnisse

11 páginas, 3 figuras, 2 tablas. ; The present study aimed to isolate and perform molecular and phenotypic characterization of Toxoplasma gondii strains infecting Iberian pigs bred under semi-free conditions and destined for human consumption. Blood and heart tissue samples from 361 fattening pigs from 10 various herds selected in the main areas of Iberian pig production were collected at a slaughterhouse; the sera were tested for anti-T. gondii antibodies using a commercial indirect ELISA kit, and a mouse bioassay was carried out using heart muscle of seropositive individual representatives from each geographical location. Seventy-nine (21.9%) of the 361 animals tested positive for anti-T. gondii antibodies according to the serology test. Fifteen samples of myocardial tissue were subjected to bioassay and 5 isolates (TgPigSp1 to TgPigSp5) were obtained. The isolates were characterized by using 11 PCR-RFLP genetic markers; three isolates had a ToxoDB #3 genotype (3/5) and two isolates had a ToxoDB #2 genotype (2/5). The TgPigSp1 and TgPigSp4 isolates were selected for virulence in mice characterization as instances of each different RFLP-genotype found. The TgPigSp1 isolate (#2 genotype) was virulent in mice with notable cumulative mortality (87.5%) and morbidity rates (100%); the TgPigSp4 (#3) was nonvirulent and triggered mild clinical signs in 42.1% of seropositive mice. Infection dynamics and organ distribution of both isolates were analyzed; the data revealed significant differences, including substantially higher parasite load in the lung during the acute phase of infection, in mice infected with TgPigSp1 than in the case of TgPigSp4 (median parasite load 7.6 vs. 0 zoites/mg, respectively; p < 0.05). Furthermore, degrees of severity of detected histopathological lesions appeared to be related to higher parasite burdens. Taking into account the unexpectedly high mortality rate and parasite load associated with the clonal genotype III, which is traditionally considered nonvirulent in mice, the need for further investigation and characterization of the T. gondii strains circulating in any host in Europe is emphasized. ; This research was supported by the projects funded by the Spanish Ministry of Science and Innovation (AGL2016- 75935-C2-R) and the Community of Madrid (PLATESA2-CMP2018/BAA-4370). MF-E was funded by UCM-Santander/2017 pre-doctoral grants. RC-B, EC-F, and LO-M are part of the TOXOSOURCES consortium supported by the funding from the European Union's Horizon 2020 Research and Innovation Programme under the grant agreement No 773830: One Health European Joint Programme.

15 páginas, 5 figuras, 2 tablas. ; Toxoplasma gondii is an important zoonotic agent with high genetic diversity, complex epidemiology, and variable clinical outcomes in animals and humans. In veterinary medicine, this apicomplexan parasite is considered one of the main infectious agents responsible for reproductive failure in small ruminants worldwide. The aim of this study was to phenotypically characterize 10 Spanish T. gondii isolates recently obtained from sheep in a normalized mouse model and in an ovine trophoblast cell line (AH-1) as infection target cells. The panel of isolates met selection criteria regarding such parameters as genetic diversity [types II (ToxoDB #1 and #3) and III (#2)], geographical location, and sample of origin (aborted foetal brain tissues or adult sheep myocardium). Evaluations of in vivo mortality, morbidity, parasite burden and histopathology were performed. Important variations between isolates were observed, although all isolates were classifed as "nonvirulent" (<30% cumulative mortality). The isolates TgShSp16 (#3) and TgShSp24 (#2) presented higher degrees of virulence. Signifcant diferences were found in terms of in vitro invasion rates and tachyzoite yield at 72 h post-inoculation (hpi) between TgShSp1 and TgShSp24 isolates, which exhibited the lowest and highest rates, respectively. The study of the CS3, ROP18 and ROP5 loci allelic profles revealed only type III alleles in ToxoDB #2 isolates and type II alleles in the #1 and #3 isolates included. We concluded that there are relevant intraand inter-genotype virulence diferences in Spanish T. gondii isolates, which could not be inferred by genetic charac‑terization using currently described molecular markers. ; This research was supported by projects funded by the Spanish Ministry of Science and Innovation (AGL2016-75935-C2-R) and the Community of Madrid (PLATESA2-CMP2018/BAA-4370). MF-E was funded by UCM-Santander/2017 pre-doctoral Grants. RC-B, EC-F, and LO-M are part of the TOXOSOURCES con‑ sortium supported by the funding from the European Union's Horizon 2020 Research and Innovation Programme under the Grant Agreement No 773830: One Health European Joint Programme.

5 páginas, 3 figuras, 2 tablas. ; Neospora caninum is one of the more-efficient transplacentally-transmitted organisms. The goal of the present study was to investigate the pathologic and immunologic changes that occur at the materno-fetal interphase in pregnant BALB/c mice infected with N. caninum at mid-gestation. Parasite DNA was detected in feto-placentary units 3 days post-infection (PI). On day 7 PI, the DNA detection level and parasite burden were significantly higher in the placentas than in the fetuses, which may indicate that the parasite is mainly multiplying in the placenta during the initial infection. In the spleens of infected dams, we observed an increase in IFN-gamma, IL-10, and IL-4. However, only IL-4 was upregulated in placentas from the infected dams; this may enhance susceptibility to N. caninum at the materno-fetal interphase and favor transmission to the progeny. Finally, an increase in TNF-alpha expression in nested-PCR-positive placentas combined with necrosis may compromise the viability of the fetuses. ; This work has been supported by a grant from the Spanish Government (AGL2001-1362). Inmaculada C. López-Pérez has been financed by the Spanish Ministry of Education and Science. This experiment complied with the current local animal protection laws of the European Union. ; Peer reviewed

18 páginas, 5 figuras, 4 tablas. ; Neospora caninum is an apicomplexan parasite that can cause abortions and perinatal mortality in sheep. Although ovine neosporosis has been described worldwide, there is a lack of information about the relationship between N. caninum serostatus and the reproductive performance. In this study, we described the infection dynamics in a dairy sheep flock with an abortion rate up to 25% and a N. caninum seroprevalence of 32%. Abortions were recorded in 36% and 9% of seropositive and seronegative sheep, respectively. Seropositive sheep were more likely to abort twice (OR = 4.44) or three or more times (OR = 10.13) than seronegative sheep. Endogenous transplacental transmission was the main route of transmission since 86% of seropositive sheep had seropositive offspring. Within dams that had any abortion, seropositive sheep were more likely than seronegative ones to have female descendants that aborted (OR = 8.12). The slight increase in seropositivity with the age, the low percentage of animals with postnatal seroconversion or with low avidity antibodies, and the seropositivity of one flock dog, indicated that horizontal transmission might have some relevance in this flock. A control programme based on selective culling of seropositive sheep and replacement with seronegative animals was effective in reducing the abortion rate to 7.2%. ; This study was funded by the Spanish Ministry of Science and Innovation (PID2019- 104713RB-C21). R.S.-S. was funded by the Community of Madrid, Spain (PLATESA2, S2018/BAA4370) and M.F.-E. by UCM-Santander/2017 predoctoral grants. L.M. Ortega-Mora and G. AlvarezGarcia are part of the TOXOSOURCES consortium, supported by funding from the European Union's Horizon 2020 Research and Innovation programme under grant agreement No 773830: One Health European Joint Programme. ; Peer reviewed