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.

Coccidiosis is a well-known poultry disease that causes the severe destruction of the intestinal tract, resulting in reduced growth performance and immunity, disrupted gut homeostasis and perturbed gut microbiota. Supplementation of probiotics were explored to play a key role in improving growth performance, enhancing innate and adaptive immunity, maintaining gut homeostasis and modulating gut microbiota during enteric infection. This study was therefore designed to investigate the chicken gut whole microbiota responses to Bacillus subtilis (B. subtilis) probiotic feeding in the presence as well as absence of Eimeria infection. For that purpose, 84 newly hatched chicks were assigned into four groups, including (1) non-treated non-challenged control group (CG − ET), (2) non-treated challenged control group (CG + ET), (3) B. subtilis-fed non-challenged group (BS − ET) and (4) B. subtilis-fed challenged group (BS + ET). CG + ET and BS + ET groups were challenged with Eimeria tenella (E. tenella) on 21 day of housing. Our results for Alpha diversity revealed that chickens in both infected groups (CG + ET and BS + ET) had lowest indexes of Ace, Chao 1 and Shannon, while highest indexes of Simpson were found in comparison to non-challenged groups (CG − ET and BS − ET). Firmicutes was the most affected phylum in all experimental groups following Proteobacteria and Bacteroidota, which showed increased abundance in both non-challenged groups, whereas Proteobacteria and Bacteroidota affected both challenged groups. The linear discriminant analysis effect size method (lEfSe) analysis revealed that compared to the CG + ET group, supplementation of probiotic in the presence of Eimeria infection increased the abundance of some commensal genera, included Clostridium sensu stricto 1, Corynebacterium, Enterococcus, Romboutsia, Subdoligranulum, Bacillus, Turicibacter and Weissella, with roles in butyrate production, anti-inflammation, metabolic reactions and the modulation of protective pathways against pathogens. Collectively, these findings evidenced that supplementation of B. subtilis probiotic was positively influenced with commensal genera, thereby alleviating the Eimeria-induced intestinal disruption.

In this study, we performed transcriptome analysis in the cecum tissues of negative control untreated non-challenged (NC), positive control untreated challenged (PC), and Bacillus subtilis (B. subtilis) fed challenged chickens (BS + ET) in order to examine the underlying potential therapeutic mechanisms of Bacillus based probiotic feeding under an experimental Eimeria tenella (E. tenella) infection. Our results for clinical parameters showed that birds in probiotic diet decreased the bloody diarrhea scores, oocyst shedding, and lesion scores compared to positive control birds. RNA-sequencing (RNA-seq) analysis revealed that in total, 2509 up-regulated and 2465 down-regulated differentially expressed genes (DEGs) were detected in the PC group versus NC group comparison. In the comparison of BS + ET group versus PC group, a total of 784 up-regulated and 493 down-regulated DEGs were found. Among them, several DEGs encoding proteins involved in immunity, gut barrier integrity, homeostasis, and metabolism were up-regulated by the treatment of probiotic. Functional analysis of DEGs also revealed that some gene ontology (GO) terms related with immunity, metabolism and cellular development were significantly affected by the exposure of probiotic. Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that the DEGs in the cecum of B. subtilis-fed challenged group were mainly participated in the pathways related with immunity and gut barrier integrity, included mitogen-activated protein kinase (MAPK) signaling pathway, toll-like receptor (TLR) signaling pathway, extracellular matrix (ECM)–receptor interaction, tight junction, and so on. Taken together, these results suggest that Bacillus based probiotic modulate the immunity, maintain gut homeostasis as well as barrier system and improve chicken metabolism during E. tenella infection.

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Semiaquilegia adoxoides (DC.) Makino is a herbal medicine and it is recorded that its water extract can be used to treat acute diseases caused by bacterial infections. In order to understand the polysaccharide of Semiaquilegia adoxoides (DC.) Makino (SMP), FT-IR and HPLC methods were performed to determine the basic chemical structure and monosaccharide compositions of SMP. The antioxidant capacity of SMP was analyzed by monitoring both the scavenging rate of DPPH and ABTS free radical. To investigate the effects of SMP on the acute bacterial disease, minimum inhibitory concentrations (MICs) of SMP on E. coli or S. aureus were detected; meanwhile, mice were administrated with SMP for 7 days and then infected with E. coli or S. aureus, and the parameters were measured at the 9th day. Results showed that SMP was a furanose which was mainly composed of glucose (60.3%) and had certain antioxidant activities. Both MIC values of SMP on E. coli and S. aureus were 250 ml/mL, which means that SMP has no direct antibacterial effects. The mice experiments revealed that SMP had potential effects on immunomodulatory by reducing WBC and the expression of serum IL-1, IL-6, and TNF-α and increasing IgM of E. coli or S. aureus infected mice. These findings supported the effect of Semiaquilegia adoxoides (DC.) Makino in folk use with scientific evidence.

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.

Litsea cubeba L. essential oil (LCEO) can affect the growth of drug-resistance bacteria. However, research on stress response of drug-resistant A. baumannii under sub-lethal LCEO concentrations had been limited so far. Therefore, transcriptomic analysis of A. baumannii under 1/2 minimum inhibitory concentration (MIC, 0.54 mg/mL) of LCEO was performed. Results of transcriptomic analysis showed that 320/352 genes were significantly up/down-regulated, respectively, in LCEO-treated A. baumannii. Both up and down-regulated genes were significantly enriched in three GO terms (oxidation-reduction process; oxidoreductase activity; oxidoreductase activity, acting on the CH-CH group of donors), which indicated that the redox state of A. baumannii was significantly affected by LCEO. LCEO may also inhibit aerobic respiration, synthesis of ketone bodies and the metabolism of some amino acids while, meanwhile, promoting fatty acid degradation of A. baumannii according to Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. The permeability and the stress of cell membrane of A. baumannii were significantly affected by LCEO. After crystal violet dyeing, the biofilm formation of A. baumannii was promoted/inhibited by extremely low/relatively high concentration of LCEO, respectively. LCEO and chloramphenicol have synergistic growth inhibitory effect against A. baumannii according to the Fractional Inhibitory Concentration Index (FICI) value = 0.375. Our results indicate that the growth of A. baumannii was inhibited by LCEO, and give insights into the stress response of A. baumannii under sub-lethal concentrations of LCEO. These results provided evidence that A. baumannii was inhibited by LCEO, and expanded knowledges of stress response of A. baumannii under sub-lethal concentration of LCEO.