Host-pathogen interaction in Toxoplasma gondii-infected mixed chicken blood cell cultures.

Toxoplasma gondii has the ability to infect various nucleated cell types in different hosts. The aim of the present study was to investigate which chicken blood cells were targeted by T. gondii in a mixed blood cell culture similar to in vivo conditions and to evaluate parasite-host cell interactions. The study consisted of two subsequent experiments. In experiment 1, we applied T. gondii tachyzoites (ME49) at a multiplicity of infection of 1 tachyzoite per blood cell and examined parasite replication, cytokine, and inducible nitric oxide synthase (iNOS) mRNA expression between 1 h and 48 h post-infection (p.i.) by quantitative PCR. By using T. gondii RH-GFP tachyzoites expressing the green fluorescent protein (GFP) in experiment 2, we aimed for visualizing infected cells by confocal laser scanning microscopy (CLSM) and flow cytometric analysis at 24 h p.i. The parasite replication curve showed a massive decrease of parasite stages until 24 h p.i. followed by an approximately plateau phase. We observed mainly significantly increased iNOS mRNA expression levels in T. gondii-infected culture compared to uninfected cells. Flow cytometry and CLSM data confirmed monocytes/macrophages as main target cells for T. gondii. Moreover, different lymphocytes like B cells and cytotoxic T cells seem to be targeted to a low extent. Our findings indicate that monocytes/macrophages play a key role during T. gondii infection in chicken as host cells and triggering of immune response. To the best of our knowledge, this is the first report of a mixed chicken blood cell culture experimentally infected with T. gondii.

Efficacy of an injectable toltrazuril - gleptoferron (Forceris®) to control coccidiosis (Cystoisospora suis) in comparison with iron supplemented piglets without anticoccidial treatment.

The efficacy and safety of an injectable combination product containing toltrazuril and gleptoferron (Forceris® - CEVA) for the control of coccidiosis due to Cystoisospora suis was investigated in neonatal piglets. The study was carried out on five European commercial pig farms in France, Germany and Spain and 122 litters consisting of 1508 piglets were selected and randomly allocated to one of two treatment groups. The first group received a single intramuscular injection per piglet of the test product, containing 45 mg toltrazuril and 200 mg iron and this was compared with a control group, which was administered a single intramuscular treatment of iron at 200 mg per piglet only. Body weights, faecal scores and oocysts counts, recorded as oocysts per gram of faeces, (OPG) were observed for 21 days. Only 1138 piglets were actually exposed to coccidiosis and data sets of these animals were selected for statistical analysis. The efficacy of the test product in the control of coccidiosis was shown in higher body weight gains, a lower percentage of animals with diarrhoea, fewer samples with positive oocysts counts as well as a lower excretion peak and a reduced area under the curve of OPG from study day (SD) 4 - SD 21. The combination product of toltrazuril and gleptoferron provided an effective alternative approach to current conventional separate treatment for the prevention of iron deficiency anaemia and coccidiosis in neonatal piglets. It reduced the numbers of potentially stressful interventions and work time.

Efficacy of an injectable toltrazuril - gleptoferron (Forceris®) to control coccidiosis (Cystoisospora suis) in comparison with iron supplemented piglets without anticoccidial treatment.

The efficacy and safety of an injectable combination product containing toltrazuril and gleptoferron (Forceris® - CEVA) for the control of coccidiosis due to Cystoisospora suis was investigated in neonatal piglets. The study was carried out on five European commercial pig farms in France, Germany and Spain and 122 litters consisting of 1508 piglets were selected and randomly allocated to one of two treatment groups. The first group received a single intramuscular injection per piglet of the test product, containing 45 mg toltrazuril and 200 mg iron and this was compared with a control group, which was administered a single intramuscular treatment of iron at 200 mg per piglet only. Body weights, faecal scores and oocysts counts, recorded as oocysts per gram of faeces, (OPG) were observed for 21 days. Only 1138 piglets were actually exposed to coccidiosis and data sets of these animals were selected for statistical analysis. The efficacy of the test product in the control of coccidiosis was shown in higher body weight gains, a lower percentage of animals with diarrhoea, fewer samples with positive oocysts counts as well as a lower excretion peak and a reduced area under the curve of OPG from study day (SD) 4 - SD 21. The combination product of toltrazuril and gleptoferron provided an effective alternative approach to current conventional separate treatment for the prevention of iron deficiency anaemia and coccidiosis in neonatal piglets. It reduced the numbers of potentially stressful interventions and work time.

Mutual interactions of the apicomplexan parasites Toxoplasma gondii and Eimeria tenella with cultured poultry macrophages.

BACKGROUND: Toxoplasma gondii and Eimeria tenella are two common parasites in poultry. Mixed infections are likely to occur frequently in chickens due to the high prevalence of both pathogens. In this study, we investigate the co-occurrence of the two pathogens in the same immunocompetent host cell population towards potential parasite-parasite as well as altered patterns of parasite-host interactions. METHODS: Primary macrophages from chicken blood were co-infected in vitro with T. gondii tachyzoites (RH strain) and E. tenella sporozoites (Houghton strain) for 72 h. Morphological observations by light microscopy and assessments of parasite replication by quantitative real-time PCR (qPCR) were performed at 24, 48 and 72 h post-infection (hpi). Six host cell immune factors previously linked to T. gondii or E. tenella infection were selected for gene expression analysis in this study. RESULTS: Distinct morphological changes of macrophages were observed during mixed infection at 24 hpi and immunological activation of host cells was obvious. Macrophage mRNA expression for iNOS at 48 hpi and for TNF-α at 72 hpi were significantly elevated after mixed infection. Distinct upregulation of IL-10 was also present during co-infection compared to Eimeria mono-infection at 48 and 72 hpi. At 72 hpi, the total number of macrophages as well as the number of both parasites decreased markedly. As measured by qPCR, E. tenella population tended to increase during T. gondii co-infection, while T. gondii replication was not distinctly altered. CONCLUSIONS: Mutual interactions of T. gondii and E. tenella were observed in the selected co-infection model. The interactions are supposed to be indirect considering the observed changes in host cell metabolism. This study would thus help understanding the course of co-infection in chickens that may be relevant in terms of veterinary and zoonotic considerations.

Experimental Toxoplasma gondii and Eimeria tenella co-infection in chickens.

The widespread apicomplexan parasites Toxoplasma gondii (T. gondii) and Eimeria tenella (E. tenella) are important pathogens with high prevalence in poultry. The aim of our study was the investigation of mutual influences in co-infected chickens, focusing on immune response and course of infection. Two separate trials were performed using in total 96 1-day-old chickens, divided into four study groups: group NC (negative control, uninfected), group PC-T (oral or intramuscular infection with T. gondii oocysts (trial 1) or tachyzoites (trial 2), respectively), group PC-E (oral infection with E. tenella (trial 1) or E. tenella and Eimeria acervulina (trial 2)), and group TE (co-infection). T. gondii and Eimeria infections were validated by different parameters, and cytokine expression in the gut and spleen was investigated. T. gondii-specific antibodies were detected earliest 4 days post infection (p.i.) by immunoblot and direct DNA detection was possible in 22.1% of all tissue samples from infected chickens. Eimeria spp. merogony seemed to be enhanced by co-infection with T. gondii, interestingly without marked differences in oocyst excretion between co-infected and Eimeria spp. mono-infected chickens. An increase of messenger RNA (mRNA) expression of Th1- (IFN-γ, IL-12, TNF-α) and Th2-related cytokines (IL-10) mainly in groups PC-E and TE was observed, however, without statistically significant differences between co-infection and single infection with Eimeria. In conclusion, most of the measurable immune response could be attributed to Eimeria infection. To the best of our knowledge, this is the first report on co-infection experiments of T. gondii with Eimeria spp. in chickens.