Table S1. Hematological parameters and white blood cell counts determined on study days 1 and 29.... more Table S1. Hematological parameters and white blood cell counts determined on study days 1 and 29. Abbreviations: MCV, mean cell volume; MCH, mean cellular hemoglobin; MCHC, mean cellular hemoglobin concentration. Table S2. Differences in hematological parameters and white blood cell counts between groups and over time. Values of SD 1 and SD 29 were compared between uninfected animals and infected animals altogether. Additionally, differences over time were calculated for both groups. Abbreviations: x ¯ $$ \overline{\mathrm{x}} $$ , mean; σ, standard deviation. Table S3. Serum titer development over time within groups. In brackets: test statistic χ2 and degrees of freedom df according to the Friedman rank sum test. Abbreviations: SD, study day; x ¯ $$ \overline{\mathrm{x}} $$ , mean titer; σ, standard deviation. Table S4. Results (as P-values) of antibody titer comparisons between groups on each study day. Significant differences (P ≤ 0.05) are indicated in bold. Abbreviations: SD, s...
To date, investigations on the immune response to Cystoisospora suis infections focused on suckli... more To date, investigations on the immune response to Cystoisospora suis infections focused on suckling piglets, the age group clinically most affected. Actively immunizing piglets is unfeasible due to their immature immune system and the typically early infection in the first days after birth. Therefore, understanding and possibly enhancing the immune response of immune-competent animals is the prerequisite to develop a passive immunization strategy for piglets which currently rely on very limited treatment options. To investigate antibody and cytokine responses of immune-competent animals and the impact of the oral immunization protocol on their immune response, growers with unknown previous exposure to C. suis (10-11 weeks-old) were infected one or three times with different doses (600 and 6000 or 200 and 2000, respectively) of C. suis oocysts, and compared to uninfected controls. Oocyst excretion was evaluated, and blood and intestinal mucus antibody titers were determined by IFAT. ...
In the current study the expression and ultrastructural localization of heat shock protein 70 (HS... more In the current study the expression and ultrastructural localization of heat shock protein 70 (HSP70) was analyzed by immunogold labelling of surface spreads of meiotic chromosomes from Eimeria tenella oocysts. The authors used a previously reported method that overcomes the difficulties of the resistance of Eimeria oocysts to disruption and permits the release of intact meiotic chromosomes. HSP70 was localized at the ultrastructural level using an anti-HSP70 monoclonal antibody in combination with a secondary antibody coupled to colloidal gold. Synaptonemal complexes (SCs) were visualized by means of the surface spreading technique to study both HSP70 expression and the consequences of the lack of HSP70 in the behaviour of the eimerian chromosomes during meiosis. For that purpose E. tenella oocysts were treated with quercetin, a flavonoid that is known to inhibit the synthesis of HSP70. The results showed a close association of HSP70 with the lateral elements (LEs) of the SCs. That association began at the time that SCs were formed and persisted until disassemble. Comparison between distribution of immunogold label over the SCs from non-treated and treated oocysts revealed a decreasing number of gold particles as the concentration of quercetin increased. The current results demonstrated three dose-dependent effects of the quercetin treatment of Eimeria oocysts: a reduction in the HSP70 synthesis; defects in SC formation or desynapsis, and inhibition of sporulation. HSP70, as a structural component of the SCs, may be involved in SC functions such as chromosomal pairing, recombination, or disjunction.
To date, investigations on the immune response to Cystoisospora suis infections focused on suckli... more To date, investigations on the immune response to Cystoisospora suis infections focused on suckling piglets, the age group clinically most affected. Actively immunizing piglets is unfeasible due to their immature immune system and the typically early infection in the first days after birth. Therefore, understanding and possibly enhancing the immune response of immune-competent animals is the prerequisite to develop a passive immunization strategy for piglets which currently rely on very limited treatment options. To investigate antibody and cytokine responses of immune-competent animals and the impact of the oral immunization protocol on their immune response, growers with unknown previous exposure to C. suis (10-11 weeks-old) were infected one or three times with different doses (600 and 6000 or 200 and 2000, respectively) of C. suis oocysts, and compared to uninfected controls. Oocyst excretion was evaluated, and blood and intestinal mucus antibody titers were determined by IFAT. ...
Eimeria praecox and Eimeria acervulina are two species of coccidia parasites infecting chickens, ... more Eimeria praecox and Eimeria acervulina are two species of coccidia parasites infecting chickens, which develop in the duodenum. The purpose of this study was to evaluate the pathogenicity of E. praecox and to study interactions of this coccidium with E. acervulina. The results showed that the pathogenicity of E. praecox was related to the infective dose, and that its impact on individual weight and weight gain was significant from the lowest administered dose: 5000 oocysts per bird. No morbidity was observed, even with the highest infective dose, but faecal consistency alteration was higher with increasing infective doses. No consistent lesion was observed. When E. praecox was associated with E. acervulina with a low infective dose, performance deterioration seemed to be an additional effect of the two species. However, in the case of heavy infections, signs worsened along with duration of negative impact on growth, compared to a mono-infection.
The present study reports the effects of various field anticoccidial programs on the distribution... more The present study reports the effects of various field anticoccidial programs on the distribution of Eimeria spp. in poultry litter and serum antibody titers against coccidia in broiler chickens raised on the used litters. The programs included in ovo vaccination and various medications with either chemicals, ionophores, or both. In general, serum samples from these chickens showed anticoccidial antibody titers when tested at days 7 and 14 post hatch with the peak response at day 43. Serum anticoccidial titers were highest in birds fed a non-medicated diet compared with those vaccinated or fed medicated diets. Total number of Eimeria oocysts and the composition of Eimeria spp. present in the litter samples from different treatment groups varied depending on the type of anticoccidial program. Oocyst counts in general ranged from 3.7×10(3) to 7.0×10(4) per g of litter. Importantly, both morphological and molecular typing studies revealed four major predominant Eimeria spp., E. acervul...
This study focuses on reporting events in Eimeria tenella oocysts from early to late prophase I i... more This study focuses on reporting events in Eimeria tenella oocysts from early to late prophase I in terms of RAD51 protein in association with the synaptonemal complex formed between homologous chromosomes. The aim of the study was the sequential localization of RAD51 protein, which is involved in the repair of double-strand breaks (DSBs) on the eimerian chromosomes as they synapse and desynapse. Structural Maintenance of Chromosome protein SMC3, which plays a role in synaptonemal complex formation, was labeled to identify initiation and progress of chromosome synapsis and desynapsis in parallel with the appearance and disappearance of RAD51 foci. Antibodies directed against RAD51 and cohesin subunit SMC3 proteins were labeled with either fluorescence or colloidal gold to visualize RAD51 protein foci and synaptonemal complexes. RAD51 protein localization during prophase I was studied on meiotic chromosomes spreads obtained from oocysts at different points in time after the start of sporulation. The present findings showed that foci detected with the antibody directed against RAD51 protein first appeared at the pre-leptotene stage before homologous chromosomes began pairing. Subsequently, the foci were detected in association with the lateral elements at the precise sites where synapsis were in progress. These findings lead us to suggest that in E. tenella, homologous chromosome pairing was a DSB-dependent mechanism and reinforced the participation of RAD51 protein in meiotic homology search, alignment and pairing of chromosomes.
Table S1. Hematological parameters and white blood cell counts determined on study days 1 and 29.... more Table S1. Hematological parameters and white blood cell counts determined on study days 1 and 29. Abbreviations: MCV, mean cell volume; MCH, mean cellular hemoglobin; MCHC, mean cellular hemoglobin concentration. Table S2. Differences in hematological parameters and white blood cell counts between groups and over time. Values of SD 1 and SD 29 were compared between uninfected animals and infected animals altogether. Additionally, differences over time were calculated for both groups. Abbreviations: x ¯ $$ \overline{\mathrm{x}} $$ , mean; σ, standard deviation. Table S3. Serum titer development over time within groups. In brackets: test statistic χ2 and degrees of freedom df according to the Friedman rank sum test. Abbreviations: SD, study day; x ¯ $$ \overline{\mathrm{x}} $$ , mean titer; σ, standard deviation. Table S4. Results (as P-values) of antibody titer comparisons between groups on each study day. Significant differences (P ≤ 0.05) are indicated in bold. Abbreviations: SD, s...
To date, investigations on the immune response to Cystoisospora suis infections focused on suckli... more To date, investigations on the immune response to Cystoisospora suis infections focused on suckling piglets, the age group clinically most affected. Actively immunizing piglets is unfeasible due to their immature immune system and the typically early infection in the first days after birth. Therefore, understanding and possibly enhancing the immune response of immune-competent animals is the prerequisite to develop a passive immunization strategy for piglets which currently rely on very limited treatment options. To investigate antibody and cytokine responses of immune-competent animals and the impact of the oral immunization protocol on their immune response, growers with unknown previous exposure to C. suis (10-11 weeks-old) were infected one or three times with different doses (600 and 6000 or 200 and 2000, respectively) of C. suis oocysts, and compared to uninfected controls. Oocyst excretion was evaluated, and blood and intestinal mucus antibody titers were determined by IFAT. ...
In the current study the expression and ultrastructural localization of heat shock protein 70 (HS... more In the current study the expression and ultrastructural localization of heat shock protein 70 (HSP70) was analyzed by immunogold labelling of surface spreads of meiotic chromosomes from Eimeria tenella oocysts. The authors used a previously reported method that overcomes the difficulties of the resistance of Eimeria oocysts to disruption and permits the release of intact meiotic chromosomes. HSP70 was localized at the ultrastructural level using an anti-HSP70 monoclonal antibody in combination with a secondary antibody coupled to colloidal gold. Synaptonemal complexes (SCs) were visualized by means of the surface spreading technique to study both HSP70 expression and the consequences of the lack of HSP70 in the behaviour of the eimerian chromosomes during meiosis. For that purpose E. tenella oocysts were treated with quercetin, a flavonoid that is known to inhibit the synthesis of HSP70. The results showed a close association of HSP70 with the lateral elements (LEs) of the SCs. That association began at the time that SCs were formed and persisted until disassemble. Comparison between distribution of immunogold label over the SCs from non-treated and treated oocysts revealed a decreasing number of gold particles as the concentration of quercetin increased. The current results demonstrated three dose-dependent effects of the quercetin treatment of Eimeria oocysts: a reduction in the HSP70 synthesis; defects in SC formation or desynapsis, and inhibition of sporulation. HSP70, as a structural component of the SCs, may be involved in SC functions such as chromosomal pairing, recombination, or disjunction.
To date, investigations on the immune response to Cystoisospora suis infections focused on suckli... more To date, investigations on the immune response to Cystoisospora suis infections focused on suckling piglets, the age group clinically most affected. Actively immunizing piglets is unfeasible due to their immature immune system and the typically early infection in the first days after birth. Therefore, understanding and possibly enhancing the immune response of immune-competent animals is the prerequisite to develop a passive immunization strategy for piglets which currently rely on very limited treatment options. To investigate antibody and cytokine responses of immune-competent animals and the impact of the oral immunization protocol on their immune response, growers with unknown previous exposure to C. suis (10-11 weeks-old) were infected one or three times with different doses (600 and 6000 or 200 and 2000, respectively) of C. suis oocysts, and compared to uninfected controls. Oocyst excretion was evaluated, and blood and intestinal mucus antibody titers were determined by IFAT. ...
Eimeria praecox and Eimeria acervulina are two species of coccidia parasites infecting chickens, ... more Eimeria praecox and Eimeria acervulina are two species of coccidia parasites infecting chickens, which develop in the duodenum. The purpose of this study was to evaluate the pathogenicity of E. praecox and to study interactions of this coccidium with E. acervulina. The results showed that the pathogenicity of E. praecox was related to the infective dose, and that its impact on individual weight and weight gain was significant from the lowest administered dose: 5000 oocysts per bird. No morbidity was observed, even with the highest infective dose, but faecal consistency alteration was higher with increasing infective doses. No consistent lesion was observed. When E. praecox was associated with E. acervulina with a low infective dose, performance deterioration seemed to be an additional effect of the two species. However, in the case of heavy infections, signs worsened along with duration of negative impact on growth, compared to a mono-infection.
The present study reports the effects of various field anticoccidial programs on the distribution... more The present study reports the effects of various field anticoccidial programs on the distribution of Eimeria spp. in poultry litter and serum antibody titers against coccidia in broiler chickens raised on the used litters. The programs included in ovo vaccination and various medications with either chemicals, ionophores, or both. In general, serum samples from these chickens showed anticoccidial antibody titers when tested at days 7 and 14 post hatch with the peak response at day 43. Serum anticoccidial titers were highest in birds fed a non-medicated diet compared with those vaccinated or fed medicated diets. Total number of Eimeria oocysts and the composition of Eimeria spp. present in the litter samples from different treatment groups varied depending on the type of anticoccidial program. Oocyst counts in general ranged from 3.7×10(3) to 7.0×10(4) per g of litter. Importantly, both morphological and molecular typing studies revealed four major predominant Eimeria spp., E. acervul...
This study focuses on reporting events in Eimeria tenella oocysts from early to late prophase I i... more This study focuses on reporting events in Eimeria tenella oocysts from early to late prophase I in terms of RAD51 protein in association with the synaptonemal complex formed between homologous chromosomes. The aim of the study was the sequential localization of RAD51 protein, which is involved in the repair of double-strand breaks (DSBs) on the eimerian chromosomes as they synapse and desynapse. Structural Maintenance of Chromosome protein SMC3, which plays a role in synaptonemal complex formation, was labeled to identify initiation and progress of chromosome synapsis and desynapsis in parallel with the appearance and disappearance of RAD51 foci. Antibodies directed against RAD51 and cohesin subunit SMC3 proteins were labeled with either fluorescence or colloidal gold to visualize RAD51 protein foci and synaptonemal complexes. RAD51 protein localization during prophase I was studied on meiotic chromosomes spreads obtained from oocysts at different points in time after the start of sporulation. The present findings showed that foci detected with the antibody directed against RAD51 protein first appeared at the pre-leptotene stage before homologous chromosomes began pairing. Subsequently, the foci were detected in association with the lateral elements at the precise sites where synapsis were in progress. These findings lead us to suggest that in E. tenella, homologous chromosome pairing was a DSB-dependent mechanism and reinforced the participation of RAD51 protein in meiotic homology search, alignment and pairing of chromosomes.
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