Aim In this study, we have examined the individual and combined protective mechanism of probiotic and Bidens pilosa on the performance and gut health of chickens during Eimeria tenella infection over a 29‐day experimental trial. Methods and Results A total of one hundred and fifty 1‐day‐old chickens were equally distributed into five treatment groups with three biological replicates: two groups were allocated as control groups (control group untreated unchallenged, CG and control positive untreated challenged, CPG) and three groups were fed diets with probiotic (PG), B. pilosa (BPG) and probiotic + B. pilosa (PG + BPG) and challenged with E. tenella. Birds of all groups were assessed for pre and post‐infection body weights, oocysts shedding, caecal lesion scores and mRNA expression levels of apoptosis related proteins (Bcl‐2, Bax and caspase‐3), antioxidant enzymes (CAT and SOD 1), pro‐inflammatory cytokines (IL‐6 and IL‐8) and tight junction proteins (CLDN 1 and ZO 1). Our results revealed that during infection (day 21–29), E. tenella challenged chickens significantly decreased the body weight compared with uninfected control chickens; however, there was no significant effect on body weight of chickens fed with probiotic, B. pilosa and probiotic + B. pilosa was observed. Eimeria tenella challenged untreated birds increased (P < 0·05) oocysts shedding, destructive ratio of caeca and mortality as compared to treated challenged birds. CPG group up‐regulated the mRNA expression levels of anti‐apoptosis protein Bcl‐2 while down‐regulated the pro‐apoptosis protein Bax relative to PG, BPG and PG + BPG groups. Moreover chickens fed probiotic, B. pilosa and probiotic + B. pilosa diets enhanced the activities of antioxidant enzymes, pro‐inflammatory cytokines and tight junction proteins with the comparison of control positive untreated challenged chickens. Conclusion These findings elaborated that feed supplementation of probiotic and B. pilosa (individually or in combination) appeared to be effective in inhibiting the occurrence of disease and decreasing the severity of Eimeria infection in chickens. Significance and Impact of the Study This study explained the underlying anti‐coccidial mechanism in which probiotic and B. pilosa (individually and/or in combination) improve the performance of chicken and protect against gut inflammatory responses caused by E. tenella.
Background Probiotics and Medicinal plants are widely used as an alternative to treat different kinds of the acute and chronic diseases in all over the world. Bidens pilosa is commonly used as foods and therapeutics against various pathogens including protozoan. In the present study probiotics and Bidens pilosa were used to explore the anticoccidial activity during experimental infection of Eimeria tenella ( E. tenella ) in chicken Methods One hundred and fifty one-day-old Chinese yellow breed chickens were divided into five equal groups. Group 1 was kept as control group (CG; un-treated, unchallenged). All other groups were inoculated with E. tenella sporulated oocysts on day 21 of age. Group 2 was kept as positive control (CPG; un-treated, challenged). Group 3 was treated with probiotics (PG) at the rate of 1g/kg of feed. Group 4 was treated with Bidens pilosa (BPG) at the dose of 0.5%/kg of feed. Group 5 was kept as probiotics + Bidens pilosa group (PG + BPG). All birds were weighed initially, day of infection and the final day of experiment. On day 5, 6, 7 and 8 post-infection (PI), bloody diarrhea was scored as normal (0) to sever (1, 2, 3 and 4), at the same time oocysts shedding was quantified by McMaster method. Gross caecal lesion scoring was checked at the final day of experiment. The apoptosis related gene expressions (Bcl-2, Bax, Caspase-3), anti-oxidant enzymatic expressions (CAT, SOD 1), Peptide transporter 1 (PepT 1), pro-inflammatory cytokines (IL-6, IL-8) and tight junction proteins expressions (CLDN 1, ZO 1) were checked by real time PCR. All related gene expressions average relative to β-actin was calculated by 2−ΔΔCt method. Results Birds treated with probiotics, Bidens pilosa and probiotics + Bidens pilosa slightly increased the body weight gain, significantly reduced oocysts shedding, destructive ratio of ceaca and mortality % than control positive group. CPG birds up-regulated the expressions of Bcl-2 while down-regulated the expressions of Bax and caspase-3 as compared to PG, BPG and PG + BPG groups. Moreover, Eimeria challenged chickens supplemented with probiotics, Bidens pilosa alone and combined showed higher anti-oxidant enzymatic activity than those of challenged un-treated chickens. Additionally, the expressions of pro-inflammatory cytokines and tight junction related proteins were also up-regulated by the treated challenged than un-treated challenged chickens. Conclusion Present results revealed that, feed supplementation of probiotics and Bides pilosa induced apoptosis, enhanced anti-oxidant enzymatic activity and pro-inflammatory cytokines as well as tight junction proteins and reduce mortality during E.tenella infection in chickens.
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