Selective breeding of cold-tolerant black soldier fly (Hermetia illucens) larvae: Gut microbial shifts and transcriptional patterns.

The larvae of black soldier fly (BSFL) convert organic waste into insect proteins used as feedstuff for livestock and aquaculture. BSFL production performance is considerably reduced during winter season. Herein, the intraspecific diversity of ten commercial BSF colonies collected in China was evaluated. The Bioforte colony was subjected to selective breeding at 12 °C and 16 °C to develop cold-tolerant BSF with improved production performance. After breeding for nine generations, the weight of larvae, survival rate, and the dry matter conversion rate significantly increased. Subsequently, intestinal microbiota in the cold-tolerant strain showed that bacteria belonging to Morganella, Dysgonomonas, Salmonella, Pseudochrobactrum, and Klebsiella genera were highly represented in the 12 °C bred, while those of Acinetobacter, Pseudochrobactrum, Enterococcus, Comamonas, and Leucobacter genera were significantly represented in the 16 °C bred group. Metagenomic revealed that several animal probiotics of the Enterococcus and Vagococcus genera were greatly enriched in the gut of larvae bred at 16 °C. Moreover, bacterial metabolic pathways including carbohydrate, lipid, amino acids, and cofactors and vitamins, were significantly increased, while organismal systems and human diseases was decreased in the 16 °C bred group. Transcriptomic analysis revealed that the upregulated differentially expressed genes in the 16 °C bred groups mainly participated in Autophagy-animal, AMPK signaling pathway, mTOR signaling pathway, Wnt signaling pathway, FoxO signaling pathway, Hippo signaling pathway at day 34 under 16 °C conditions, suggesting their significant role in the survival of BSFL. Taken together, these results shed lights on the role of intestinal microflora and gene pathways in the adaptation of BSF larvae to cold stress.

Inflammasome activation by viral infection: mechanisms of activation and regulation.

Viral diseases are the most common problems threatening human health, livestock, and poultry industries worldwide. Viral infection is a complex and competitive dynamic biological process between a virus and a host/target cell. During viral infection, inflammasomes play important roles in the host and confer defense mechanisms against the virus. Inflammasomes are polymeric protein complexes and are considered important components of the innate immune system. These immune factors recognize the signals of cell damage or pathogenic microbial infection after activation by the canonical pathway or non-canonical pathway and transmit signals to the immune system to initiate the inflammatory responses. However, some viruses inhibit the activation of the inflammasomes in order to replicate and proliferate in the host. In recent years, the role of inflammasome activation and/or inhibition during viral infection has been increasingly recognized. Therefore, in this review, we describe the biological properties of the inflammasome associated with viral infection, discuss the potential mechanisms that activate and/or inhibit NLRP1, NLRP3, and AIM2 inflammasomes by different viruses, and summarize the reciprocal regulatory effects of viral infection on the NLRP3 inflammasome in order to explore the relationship between viral infection and inflammasomes. This review will pave the way for future studies on the activation mechanisms of inflammasomes and provide novel insights for the development of antiviral therapies.

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.

Chicken Gut Microbiota Responses to Dietary Bacillus subtilis Probiotic in the Presence and Absence of Eimeria Infection.

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.

Immunomodulatory and ameliorative effects of probiotic in combination with diclazuril on broilers under coccidia infection.

AIM: This study aimed to determine the potential prophylactic efficacy of probiotic individually and/or in combination with anti-coccidial drug on the performance and immunity of broilers under an induced coccidial infection over a 28-day of experimental trial. METHODS: One hundred and eighty 1-day-old Cobb broiler chicks were randomly divided into five groups, included control group (CG), control positive group (CPG), probiotic-treated group (Prob), diclazuril-treated group (Dic), and probiotic + diclazuril-treated group (Prob + Dic). On day 21 of age, all birds, except group CG, were orally inoculated with 1 ml of tap water containing 25,000 Eimeria tenella sporulated oocysts. RESULTS: Our results showed that the probiotic treatment did not influence pre-challenge body weight, feed intake and feed conversion ratio (FCR). During the post-challenge period, chickens in groups probiotic and diclazuril individually and in combination exhibited higher body weight and lower (better) FCR, reduced oocyst shedding (throughout the day four, five, six and seven post-infection), cecal lesions and mortality compared with control positive chickens. Moreover, Compared to CPG group, Prob + Dic group showed increased (p < 0.05) serum levels of interleukin-10 (IL-10) and immunoglobulin M (IgM) and decreased the concentrations of interferon gamma (IFN-γ). On the other hand, individual treatment with probiotic exhibited highest serum levels of IL-10 and IgM, while diclazuril alone increased the blood concentrations of IL-10 and decreased the levels of IFN-γ compared to control positive group; however, there was no significant effect of Prob on IFN-γ, Dic on IgM and all groups on interleukin-17. CONCLUSION: In conclusion, supplementation of probiotic, with and/or without anti-coccidial drug, enhances immunity and inhibits the negative effects of Eimeria infection. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reveals the anti-coccidial mechanisms of probiotic in the presence and absence of anti-coccidial drug in preventing the coccidia infection.

A Natural Antimicrobial Agent: Analysis of Antibacterial Effect and Mechanism of Compound Phenolic Acid on Escherichia coli Based on Tandem Mass Tag Proteomics.

The objective of this study was to evaluate the antibacterial mechanisms of phenolic acids as natural approaches against multi-drug resistant Escherichia coli (E. coli). For that purpose, five phenolic acids were combined with each other and 31 combinations were obtained in total. To select the most potent and effective combination, all of the obtained combinations were examined for minimum inhibitory concentration (MIC) and it was found that the compound phenolic acid (CPA) 19 (protocatechuic acid, hydrocinnamic acid, and chlorogenic acid at concentrations of 0.833, 0.208, and 1.677 mg/mL, respectively) showed better efficacy against E. coli compared to other combinations. Furthermore, based on tandem mass tag (TMT) proteomics, the treatment of CPA 19 significantly downregulated the proteins associated with resistance (Tsr, Tar, CheA, and CheW), OmpF, and FliC of multidrug-resistant E. coli. At the same time, we proved that CPA 19 improves the sensitivity of E. coli to antibiotics (ceftriaxone sodium, amoxicillin, fosfomycin, sulfamonomethoxine, gatifloxacin, lincomycin, florfenicol, cefotaxime sodium, and rifampicin), causes the flagellum to fall off, breaks the structure of the cell wall and cell membrane, and leads to macromolecules leaks from the cell. This evidence elaborated the potential therapeutic efficacy of CPA 19 and provided a significant contribution to the discovery of antibacterial agents.

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.

Transcriptome Analysis Revealed Ameliorative Effects of Bacillus Based Probiotic on Immunity, Gut Barrier System, and Metabolism of Chicken under an Experimentally Induced Eimeria tenella Infection.

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.