Citric Acid and Magnolol Ameliorate Clostridium perfringens Challenge in Broiler Chickens.

Clostridium perfringens (C. perfringens) is a common pathogenic bacterium implicated in the enteric diseases of animals. Each year, the disease is responsible for billions of dollars of losses worldwide. The development of new phytomedicines as alternatives to antibiotics is becoming a new hotspot for treating such diseases. Citric acid (CA) and magnolol (MA) have been shown to have antibacterial, antioxidant, and growth-promoting properties. Here, the bacteriostatic effects of combinations of CA and MA against C. perfringens were investigated, together with their effects on yellow-hair chickens challenged with C. perfringens. It was found that the optimal CA:MA ratio was 50:3, with a dose of 265 µg/mL significantly inhibiting C. perfringens growth, and 530 µg/mL causing significant damage to the bacterial cell morphology. In animal experiments, C. perfringens challenge reduced the growth, damaged the intestinal structure, activated inflammatory signaling, impaired antioxidant capacity, and perturbed the intestinal flora. These effects were alleviated by combined CA-MA treatment. The CA-MA combination was found to inhibit the TLR/Myd88/NF-κB and Nrf-2/HO-1 signaling pathways. In conclusion, the results suggest the potential of combined CA-MA treatment in alleviating C. perfringens challenge by inhibiting the growth of C. perfringens and affecting the TLR/MyD88/NF-κB and Nrf-2/HO-1 signaling pathways.

Punicalagin: a monomer with anti-Eimeria tenella effect from fruit peel of Punica granatum L.

Poultry production was long plagued by coccidiosis, and the development of alternative therapies will make practical sense. In this work, 2 battery experiments were designed. In battery experiment 1, the best effect of 7 anticoccidial herbs (Sophora japonica Linn, Citrus aurantium L, leaf of Acer palmatum, bark of Magnolia officinalis, fruit peel of Punica granatum L., Eclipta prostrata L., and Piper sarmentosum Roxb.) against Eimeria tenella infection of 21-day-old male Chinese Guangxi yellow-feathered chickens were screened out by clinic indexes (bloody feces scores, cecal lesion scores, oocysts output, relative weight gain rate, and survival rate). According to the results from battery experiment 1 and other literature research, we selected 2 monomers which were extracted from fruit peel of Punica granatum L. for further battery experiment 2 which were similar with battery experiment 1. Clinic results showed that Punicalagin had better anticoccidial effect than Ellagic acid. The anticoccidial mechanism exploration results of Elisa, antioxidant test, and pathological observation showed that Punicalagin reduced the cecal inflammation, improved the expression of immunoglobulin in cecal tissue, improved cecal integrity, and restored its REDOX state. Results of 16S rRNA sequencing analysis showed that Punicalagin also maintained the fecal flora health during E. tenella infection through insignificantly increasing the proportion of Lactobacillus and Faecalibacterium as well as significantly reducing the proportion of pathogenic bacteria, Escherichia-Shigella. RNA-Seq analysis results suggested that Punicalagin may play a role in controlling E. tenella infection by interaction with cytochrome P450 family enzymes. Overall, Punicalagin has promising potential as an alternative therapy for chicken Eimeria tenella infection.

Dietary supplementation with Piper sarmentosum extract on gut health of chickens infected with Eimeria tenella.

To explore the potential alternative of anti-coccidials, we investigated the therapeutic efficacy of dietary Piper sarmentosum extract (PSE) on induced coccidia infection in chickens. A total of 96-day-old chickens were randomly distributed to 1 of 3 treatment groups, including (1) control negative untreated uninfected (CN), (2) control positive untreated infected (CP), and (3) Piper sarmentosum (P. sarmentosum) extract-treated infected group (PSE). Our results demonstrated that E. tenella challenged untreated group showed a reduction (P < 0.05) in post-infection (PI) body weight compared to control negative group. However, supplementation of P. sarmentosum extract had no significant effects on body weight and cecal lesions compared with control positive group. Infected chickens fed PSE diet decreased (P < 0.05) the bloody diarrhea scores and oocyst shedding (during the day 5 to 8 post-infection) than that of CP chickens. E. tenella-challenged chickens upregulated (P < 0.05) the mRNA expression of IL-8 and Bcl-2 compared to PSE chickens, while IFN-γ compared to CN chickens. On the other hand, treatment of P. sarmentosum extract tended to increase (P < 0.05) the transcription patterns of IL-4, IL-10, CLDN 1, SOD 1, and Bax with the comparison of control positive group; however, there were no significant effects on IL-8, ZO 1, and CAT expression between the PSE and CP groups. Collectively, these findings elaborated that dietary P. sarmentosum extract exhibit potential anti-coccidial effects in controlling the coccidia infection in chickens.

Transcriptomic Analysis of Drug-Resistance Acinetobacter baumannii under the Stress Condition Caused by Litsea cubeba L. Essential Oil via RNA Sequencing.

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 analysisof 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.

The Antioxidant Polysaccharide from Semiaquilegia adoxoides (DC.) Makino Adjusts the Immune Response of Mice Infected by Bacteria.

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.