The inhibition mechanism of PostbioYDFF-3 on quality deterioration of refrigerated grass carp fillets from the perspective of endogenous enzyme and microorganisms changes.

The protective mode of PostbioYDFF-3 (referred to as postbiotics) on the quality stability of refrigerated fillets was explored from the aspects of endogenous enzyme activity and the abundance of spoilage microorganisms. Compared to the control group, the samples soaked in postbiotics showed significant reductions in TVC, TVB-N and TBARS values by 39.6%, 58.6% and 25.5% on day 5, respectively. In addition, the color changes, biogenic amine accumulation and texture softening of the fish fillets soaked in postbiotics were effectively suppressed. Furthermore, the activity of endogenous enzyme activities was detected. The calpain activities were significantly inhibited (p < 0.05) after soaking in postbiotics, which declined by 23%. Meanwhile, high throughput sequencing analysis further indicated that the growth of spoilage microorganism such as Acinetobacter and Pseudomonas were suppressed. Overall, the PostbioYDFF-3 was suitable for preserving fish meat.

Bacillus subtilis HW2 enhances growth performance and alleviates gut injury via attenuation of endoplasmic reticulum stress and regulation of gut microbiota in broilers under necrotic enteritis challenge.

This study investigated the effects of Bacillus subtilis HW2 on the growth performance, immune response, endoplasmic reticulum (ER) stress, and intestinal health in broilers with necrotic enteritis. Three hundred 1-day-old male Cobb 500 broilers (33.88 ± 2.34 g) were randomly allocated to 5 groups including non-infected control (NC group), basal diet + necrotic enteritis challenge (NE group), basal diet + 1 × 106 CFU/g B. subtilis HW2 + necrotic enteritis challenge (L-Pro group), basal diet + 5 × 106 CFU/g B. subtilis HW2 + necrotic enteritis challenge (M-Pro group), and basal diet + 1 × 107 CFU/g B. subtilis HW2 + necrotic enteritis challenge (H-Pro group), with 6 replicates per group. All broilers except NC group were orally given with sporulated coccidian oocysts at day 14 and Clostridium perfringens from days 19 to 21. Results showed that L-Pro and M-Pro groups improved growth performance and intestinal morphology in necrotic enteritis-challenged broilers, and L-Pro, M-Pro, and H-Pro groups improved intestinal barrier function and immune response and decreased ER stress in necrotic enteritis-challenged broilers. Analysis of the gut microbiota revealed that L-Pro group increased the abundances of Alistipes, Coprobacter, Barnesiella, and Limosilactobacillus, decreased Erysipelatoclostridium abundance on day 42 in necrotic enteritis-challenged broilers. M-Pro group increased Turicibacter abundance on day 28 and the abundances of Alistipes, Barnesiella, and Limosilactobacillus on day 42 in necrotic enteritis-challenged broilers. H-Pro group decreased Romboutsia abundance on day 28 and unidentified_Clostridia abundance on day 42 in necrotic enteritis-challenged broilers. Analysis of short-chain fatty acids (SCFAs) revealed higher isobutyric acid and isovaleric acid levels in L-Pro and M-Pro groups than NE group. Correlation analysis revealed the correlations between the biochemical parameters and gut microbiota as well as SCFAs, especially Romboutsia, Barnesiella, Coprobacter, isobutyric acid, and isovaleric acid. Overall, our results indicated that B. subtilis HW2 supplementation could ameliorate necrotic enteritis infection-induced gut injury. The optimal dietary supplementation dosage of Bacillus subtilis HW2 was 5 × 106 CFU/g.

Multi-omics dataset of bovine mammary epithelial cells stimulated by ten different essential amino acids.

Application of high-throughput sequencing and screening help to detect the transcriptional and metabolic discrepancies in organs provided with various levels of nutrients. The influences of individual essential amino acid (EAA) administration on transcriptomic and metabolomic profilings of bovine mammary epithelial cells (BMECs) were systematically investigated. A RNA sequencing and liquid chromatography-tandem mass spectrometry generated a comprehensive comparison of transcriptomics, non-targeted metabolomics and targeted amino acids profilings of BMECs with individual EAA stimulation by turn. The sequencing data and raw LC-MS/MS data of samples were presented in the databases of Gene Expression Omnibus, MetaboLights and Figshare for efficient reuse, including exploring the divergences in metabolisms between different EAAs and screening valuable genes and metabolites regulating casein synthesis.

CircRNA-5335 Regulates the Differentiation and Proliferation of Sheep Preadipocyte via the miR-125a-3p/STAT3 Pathway.

The content of intramuscular fat (IMF) from preadipocytes is proportional to meat quality in livestock. However, the roles of circRNAs in IMF deposition in sheep are not well known. In this study, we show that circRNA-5335/miR-125a-3p/STAT3 play a crucial adjective role in the proliferation and differentiation of sheep preadipocytes. In this study, we characterized the roles of differentially expressed circRNA-5335/miR-125a-3p/STAT3, which were screened from sheep of different months of age and based on sequencing data. Firstly, the expression profiles of circRNA-5335/miR-125a-3p/STAT3 were identified during the differentiation of preadipocytes in vitro by RT-qPCR and WB. Then, the targeting relationship of the circRNA-5335/miR-125a-3p/STAT3 was verified by dual-luciferase reporter assays. The results of RT-qPCR, CCK8, EdU and Oil Red O staining assay showed that miR-125a-3p suppressed the differentiation and raised the proliferation of preadipocytes by targeting STAT3. As a competing endogenous RNA, the downregulation of circRNA-5335 decreased the expression of STAT3 by increasing miR-125a-3p, which inhibited the differentiation of preadipocytes and promoted proliferation. Our present study demonstrates the functional significance of circRNA-5335/miR-125a-3p/STAT3 in the differentiation of sheep preadipocytes, and provides novel insights into exploring the mechanism of IMF.

Chlorogenic acid protects against intestinal inflammation and injury by inactivating the mtDNA-cGAS-STING signaling pathway in broilers under necrotic enteritis challenge.

This study aimed to determine the effects of chlorogenic acid (CGA) on the growth performance, intestinal health, immune response, and mitochondrial DNA (mtDNA)-cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) signaling pathway in broilers under necrotic enteritis (NE) challenge. The 180 one-day-old male Cobb 500 broilers with similar body weight of 44.59 ± 1.39 g were randomly allocated into 3 groups. The groups were control diet (Control group), control diet + NE challenge (NE group), and control diet + 500 mg/kg CGA + NE challenge (NE + CGA group), with 6 replicates per treatment. All broilers except the Control group were given sporulated coccidian oocysts (d 14) and Clostridium perfringens (d 19-21) by oral gavage. Our findings showed that CGA improved the growth performance and intestinal morphology in broilers under NE challenge. CGA supplementation elevated the barrier function in broilers under NE challenge, which reflected in the decreased serum concentrations of D-lactate and diamine oxidase, and upregulated jejunal protein expression of occludin. CGA supplementation also improved the immune function, which reflected in the increased concentrations and gene expressions of anti-inflammatory factors, and decreased concentrations and gene expressions of proinflammatory factors. CGA supplementation further enhanced intestinal cell proliferation and differentiation, which manifested in the increased number of goblet cells and positive cells of proliferating cell nuclear antigen on d 28 and 42. Furthermore, CGA supplementation decreased the mtDNA (d 42) and mitochondrial reactive oxygen species levels (d 28 and 42), and increased the mitochondrial membrane potential (d 42) and mitochondrial complex I (d 28 and 42) or III (d 28) activity. Broilers challenged with NE had upregulated jejunal protein expressions of cGAS, phospho-TANK-binding kinase 1, and phospho-interferon regulatory factor 7 compared with the Control group, which were downregulated after CGA supplementation. In conclusion, dietary supplementation CGA could protect against intestinal inflammation and injury by reducing the leakage of mtDNA and inactivating the cGAS-STING signaling pathway in broilers under NE challenge.

Effects of Lactobacillus plantarum HW1 on Growth Performance, Intestinal Immune Response, Barrier Function, and Cecal Microflora of Broilers with Necrotic Enteritis.

The purpose of the study was to investigate the effects of Lactobacillus plantarum HW1 on growth performance, intestinal immune response, barrier function, and cecal microflora of broilers with necrotic enteritis. In total, 180 one-day-old male Cobb 500 broilers were randomly allocated into three groups comprising a non-infected control (NC) group, basal diet + necrotic enteritis challenge (NE) group, and basal diet + 4 × 106 CFU/g Lactobacillus plantarum HW1 + necrotic enteritis challenge (HW1) group. Broilers in the NE and HW1 groups were orally given sporulated coccidian oocysts at day 14 and Clostridium perfringens from days 19 to 21. The results showed that the HW1 treatment increased (p < 0.05) the average daily gain of broilers from days 15 to 28 and from days 0 to 28 compared with the NE group. Moreover, the HW1 treatment decreased (p < 0.05) the oocysts per gram of excreta, intestinal lesion scores, ileal interleukin (IL) 1ß and tumor necrosis factor α levels, and serum D-lactic acid and diamine oxidase levels, while increasing (p < 0.05) the ileal IL-10 level, thymus index, and protein expressions of ileal occludin and ZO-1. Additionally, the HW1 treatment decreased (p < 0.05) the jejunal and ileal villus height, jejunal villus height/crypt depth value, and cecal harmful bacterial counts (Clostridium perfringens, Salmonella, Escherichia coli, and Staphylococcus aureus), and increased (p < 0.05) the cecal Lactobacillus count. In conclusion, dietary supplementation with 4 × 106 CFU/g Lactobacillus plantarum HW1 could relieve necrotic enteritis infection-induced intestinal injury and improve growth performance in broilers by improving intestinal barrier function and regulating intestinal microbiology.

Gut microbiota metabolites, redox status, and the related regulatory effects of probiotics.

Oxidative stress is a state of imbalance between oxidation and antioxidation. It is caused by excess levels of free radicals and leads to the damage of DNA, proteins, and lipids. The crucial role of gut microbiota in regulating oxidative stress has been widely demonstrated. Studies have suggested that the redox regulatory effects of gut microbiota are related to gut microbiota metabolites, including fatty acids, lipopolysaccharides, tryptophan metabolites, trimethylamine-N-oxide and polyphenolic metabolites. In recent years, the potential benefits of probiotics have been gaining increasing scientific interest owing to their ability to modulate gut microbiota and oxidative stress. In this review, we summarise the adverse health effects of oxidative stress and discuss the role of the gut microbiota and its metabolites in redox regulation. Based on the influence of gut microbiota metabolites, the roles of probiotics in preventing oxidative stress are highlighted.

Effects of Lactiplantibacillus plantarum on oxidative stress, mitophagy, and NLRP3 inflammasome activation in broiler breast meat.

Poultry meat has a high polyunsaturated fatty acids content, making it vulnerable to oxidative stress. Mitophagy participates in the regulation of oxidative stress and the nucleotide-binding and oligomerization domain (NOD)-like receptor family as well as pyrin domain-containing protein 3 (NLRP3) inflammasome activation. Lactiplantibacillus plantarum P8 (P8) is a probiotic strain with an antioxidant capacity. In the present study, we investigated the effects of P8 on oxidative stress, mitochondrial function, mitophagy, and NLRP3 inflammasome in the breast meat of oxidatively stressed broilers. Four hundred 1-day-old male broilers were assigned to a 2 × 2 factorial design with 2 P8 levels (0 or 1 × 108 cfu/g), either with or without dexamethasone (DEX) injection, for a 21-day experimental period. DEX was injected intraperitoneally once daily from d 16 to 21. The breast meat was collected on d 21. The results showed that P8 supplementation decreased malondialdehyde (MDA) levels, increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and activated the Keap1-Nrf2 pathway in DEX-injected broilers. Moreover, P8 supplementation downregulated mitochondrial DNA (mtDNA) copy number and increased the expressions of peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α), silent information regulator 1 (SIRT1), mitochondrial fusion protein 1 (Mfn1), and optic atrophy protein 1 (OPA1) in DEX-treated broilers. In addition, the decreased mitophagy level in DEX-treated broilers was elevated with P8 supplementation, as reflected by the increased gene expression of autophagy-related gene 5 (ATG5), Bcl-2-interacting protein (Becline-1), Parkin, PTEN-induced kinase 1 (PINK1), light chain 3 II (LC3II)/LC31, and the protein expression of Parkin as well as decreased p62 expression. In addition, P8 supplementation inhibited NLRP3 inflammasome activation by decreasing the transcription of NLRP3, IL-18, cysteinyl aspartate-specific proteinase-1 (Caspase-1), and the expression of NLRP3 and IL-18 in DEX-treated broilers. In conclusion, dietary P8 supplementation alleviates oxidative stress, improves mitophagy, and inhibits NLRP3 inflammasome activation in the breast meat of oxidatively stressed broilers.

Identification and analysis of differentially expressed microRNAs in endometrium to explore the regulation of sheep fecundity.

BACKGROUND: MicroRNAs (miRNAs) play an important regulatory role in mammalian reproduction. Currently, most studies are primarily concentrated on ovarian miRNAs, ignoring the influence of endometrial miRNAs on the fecundity of female sheep. To uncover potential regulators of sheep fecundity, RNA-seq was used to comparatively analyze miRNA expression profiles of endometrium between high prolificacy sheep (HP, litter size = 3) and low prolificacy sheep (LP, litter size = 1) with FecB genotype. RESULTS: Firstly, genomic features of miRNAs from endometrium were analyzed. Furthermore, 58 differentially expressed (DE) miRNAs were found in the endometrium of Hu sheep with different litter size. A co-expression network of DE miRNAs and target genes has been constructed, and hub genes related litter size are included, such as DE miRNA unconservative_NC_019472.2_1229533 and unconservative_NC_019481.2_1637827 target to estrogen receptor α (ESR1) and unconservative_NC_019481.2_1637827 targets to transcription factor 7 (TCF7). Moreover, functional annotation analysis showed that the target genes (NRCAM and NEGR1) of the DE miRNAs were significantly enriched in cell adhesion molecules (CAMs) signaling pathway, which was related to uterine receptivity. CONCLUSION: Taken together, this study provides a new valuable resource for understanding the molecular mechanisms underlying Hu sheep prolificacy.

Multi-omics reveals the mechanisms underlying Lactiplantibacillus plantarum P8-mediated attenuation of oxidative stress in broilers challenged with dexamethasone.

Oxidative stress is a common phenomenon in poultry production. Several molecules, including antioxidant genes, miRNAs, and gut microbiota metabolites, have been reported to participate in redox regulation. Lactiplantibacillus plantarum P8 (P8) was shown to improve the antioxidant capacity of chickens, but the specific molecular mechanisms remain unclear. In this study, 400 broilers were allocated to 4 treatment groups: control diet (Con group), control diet + dexamethasone injection (DEX group), control diet containing 1 × 108 CFU/g P8 (P8 group), and control diet containing 1 × 108 CFU/g P8 + DEX injection (DEX_P8 group). Integrated analysis of the microbiome, metabolomics, and miRNAomics was conducted to investigate the roles of P8 in oxidative stress in broilers. Results demonstrated that P8 supplementation significantly improved growth performance, jejunal morphology, and antioxidant function in DEX-treated broilers. Analysis of the gut microbiota revealed a higher abundance of Barnesiella (P = 0.01) and Erysipelatoclostridium (P = 0.05) in the DEX_P8 group than in the DEX group. Functional prediction indicated that certain pathways, including the phenylacetate degradation pathway, were enriched in the DEX_P8 group compared to the DEX group. Metabolites in the cecal contents were distinct between the groups. P8 supplementation increased the content of metabolites with antioxidant capacity, e.g., urobilinogen (P < 0.01), and decreased that of metabolites related to oxidative stress, e.g., genistein (P < 0.01). Functional prediction indicated that metabolites that differed between the DEX_P8 and DEX groups were enriched in pathways including "tryptophan metabolism" and "primary bile acid biosynthesis". The miRNAomics analysis further showed that, compared to the DEX group, several miRNAs in the jejunum, such as gga-miR-21-3p (P = 0.03), were increased, whereas gga-miR-455-3p (P = 0.02) was decreased in the DEX_P8 group. The PI3K-Akt, Ras, and Rap1 signaling pathways were enriched in the DEX_P8 group compared to the DEX group through KEGG analysis. Correlation analysis revealed potential interactions between growth performance, oxidation/antioxidation, jejunal morphology, gut microbiota, cecal content metabolites, and jejunal miRNAs. Overall, our results indicate that P8 supplementation may improve the growth performance, jejunal morphology and antioxidant capacity of DEX-treated broilers by regulating gut microbiota, its metabolites, and intestinal miRNAs.

Integrated multi-omics reveals the beneficial role of chlorogenic acid in improving the growth performance and immune function of immunologically stressed broilers.

Intensive production can cause immunological stress in commercial broilers. Chlorogenic acid (CGA) regulates the intestinal microbiota, barrier function, and immune function in chickens. As complex interrelations regulate the dynamic interplay between gut microbiota, the host, and diverse health outcomes, the aim of this study was to elucidate the immunoregulatory mechanisms of CGA using multi-omics approaches. A total of 240 one-day-old male broilers were assigned to a 2 × 2 factorial design with 2 CGA levels (0 or 500 mg/kg) either with or without dexamethasone (DEX) injection for a 21-day experimental period. Therefore, there were 4 dietary treatments: control, DEX, CGA, and DEX + CGA, with 6 replicates per treatment. CGA supplementation improved (P < 0.05) growth performance, jejunal morphology, jejunal barrier function, and immune function in DEX-treated broilers. Moreover, in DEX + CGA-treated broilers, the increase in gut microbiome diversity (P < 0.05) was consistent with a change in taxonomic composition, especially in the Clostridiales vadin BB60_group. Additionally, the levels of short-chain fatty acids increased remarkably (P < 0.01) after CGA supplementation. This was consistent with the Kyoto Encyclopedia of Genes and Genomes analysis results that the "pyruvate fermentation to butanoate" pathway was more enriched (P < 0.01) in the DEX + CGA group than in the DEX group. Proteomics revealed that CGA treatment increased the expression of several health-promoting proteins, thymosin beta (TMSB4X) and legumain (LGMN), which were verified by multiple reaction monitoring. Metabolomics revealed that CGA treatment increased the expression of health-promoting metabolites (2,4-dihydroxy benzoic acid and homogentisic acid). Proteomic and metabolic analyses showed that CGA treatment regulated the peroxisome proliferator-activated receptor (PPAR) and mitogen-activated protein kinase (MAPK) pathways. Western blotting results support these findings. Pearson's correlation analyses showed correlations (P < 0.01) between altered immune function, jejunal barrier function, different microbiota, proteins, and metabolites parameters. Overall, our data indicate that CGA treatment increased growth performance and improved the immunological functions of DEX-treated broilers by regulating gut microbiota and the PPAR and MAPK pathways. The results offer novel insights into a CGA-mediated improvement in immune function and intestinal health.

Hepatic Proteomics Analysis Reveals Attenuated Endoplasmic Reticulum Stress in Lactiplantibacillus plantarum-Treated Oxidatively Stressed Broilers.

Endoplasmic reticulum (ER) stress plays important roles in oxidative stress (OS), contributing to liver injury. Lactiplantibacillus plantarum P8 (P8) was reported to regulate broiler OS and the gut microbiota in broilers, but its roles in hepatic ER stress remain unclear. In the present study, the role of P8 in liver OS and ER stress was evaluated, and proteomics was performed to determine the mechanism. Results revealed that P8 treatment decreased liver OS and ER stress in dexamethasone (DEX)-induced oxidatively stressed broilers. Proteomics showed that differentially expressed proteins (DEPs) induced by DEX cover the "cellular response to unfold protein" term. Moreover, the DEPs (GGT5, TXNDC12, and SRM) between DEX- and DEX + P8-treated broilers were related to OS and ER stress and enriched in the glutathione metabolism pathway. RT-qPCR further confirmed the results of proteomics. In conclusion, P8 attenuates hepatic OS and ER stress by regulating GGT5, TXNDC12, SRM, and glutathione metabolism in broilers.

Probabilistic assessment of dietary rare earth elements intake among people living near a rare earth ore.

Rare earth elements (REEs) can cause neoplasms, reduce bone density, affect children's intelligence, etc., and diet is an important way for the human body to absorb REEs. With the increasing use of REEs, the impact on human health is becoming more and more important. So, we used a probabilistic assessment method with Monte Carlo simulation to evaluate the dietary intake of REEs by residents of a large light rare earth mining area in Shandong Province. 16 REEs in 447 samples (including wheat, maize, dry beans, vegetables, fruits and eggs) were detected. The mean value of total REEs for all samples was 286.96 µg/kg, and of light rare earth elements (LREEs) was 270.18 µg/kg. Among of LREEs, Ce, La, Nd and Pr were dominant. The REEs content of different food categories showed that wheat, leafy vegetables and allium vegetables had higher content of REEs, melons vegetables, root vegetables, fruits and eggs had the lowest content. The mean dietary intake of rare earth oxides for the whole population was 4.20 µg/kg bw/d, wheat and vegetables (leafy vegetables, allium vegetables and root vegetables) were the main sources of REEs. Dietary intake estimates of REEs by age and gender did not exceed the acceptable daily intake which means implying no impact on human health.

Pesticide residues and dietary risk assessment in radishes in Shandong.

Pesticide residues in radishes can induce serious health hazards, especially in children and toddlers. In order to assess potential health risk from pesticide residues in radishes, a total of 26 pesticides were evaluated by gas chromatography with mass spectrometry in 1690 samples, which were collected from the year 2016 to 2019 in Shandong Province of China. All the 26 pesticide residues were detected in 752 radish samples (44.50%), but only 221 samples (13.08%) contained detectable pesticide residues, which are above the maximum residue limits (MRLs). Multiple residues with two to nine pesticides were present in 5.09% (86 out of 1690) of samples. Hazard quotient (HQ) and the cumulative risk index were far below 100, while percentage value of acute reference dose (%ARfD) of triazophos exceeded 100 for adults, children, and toddlers. The %ARfD value for carbofuran, aldicarb, monocrotophos, and parathion was over 100 for toddlers. From the perspective of public health, the occurrence of pesticide residues in radishes could not pose a serious health risk problem, but the acute health risk should be paid more attention, especially to toddlers. It is recommended to make strict regulations on the management of pesticide residues and human health risk assessment about pesticide residues.

Prevalence and influence factors for myopia and high myopia in schoolchildren in Shandong, China.

OBJECTIVES: The aim of the study was to identify the prevalence and risk factors of myopia and high myopia in students from primary school and junior high school in Shandong. METHODS: A total of 35,614 subjects completed the visual acuity test, refraction error measurement, and questionnaire in 2019. The visual acuity test was performed using the standard logarithmic visual acuity chart and the refractive error was measured by an automatic refractometer without cycloplegia. RESULTS: The average age was 12.38 ± 1.78 years, with 18,501 boys and 17,113 girls. The overall prevalence of myopia and high myopia was 68.02% and 5.90%, respectively, and reached up to 85.54% and 13.13% for the grade 9 students. The risk factors included girls, parental myopic history, time spent doing homework, and less sleep time. Performing eye exercise was significantly associated with a lower risk of myopia. Use of mobile devices and reading while lying down were only related to myopia, not high myopia. CONCLUSION: The prevalence of myopia and high myopia is at a high level. In addition to genetic factors, continuous close work and a lack of sleep was an important factor associated with children myopia and high myopia.

Chlorogenic acid improves growth performance and intestinal health through autophagy-mediated nuclear factor erythroid 2-related factor 2 pathway in oxidatively stressed broilers induced by dexamethasone.

The effects of chlorogenic acid (CGA) on growth performance, intestinal morphology, antioxidant capacity, and the autophagy-mediated nuclear factor erythroid 2-related factor 2 (Nrf2) pathway in oxidatively stressed broilers were investigated. A total of 400 one-day-old male Cobb broilers were divided randomly into 4 groups using a 2 × 2 factorial arrangement with 2 CGA supplemental levels (0 and 500 mg/kg) and 2 dexamethasone (DEX) challenge levels (0 and 3 mg/kg body weight). All the broilers were injected intraperitoneally with DEX or sterile saline beginning at the age of 15 d for 6 consecutive days. The experiment lasted for 21 d. The CGA increased average daily gain (ADG), villus height, villus height/crypt depth (V/C) value, and the protein expressions of Occludin and ZO-1 in the ileum and decreased the feed:gain (F:G) ratio, which were impaired by the DEX challenge. Superoxide dismutase (SOD), catalase (CAT), gutathione S-transferase (GST), and heme oxygenase-1 (HO-1) activities in the serum and ileum were increased by CGA, whereas protein carboxyl (PCO) level in the serum and ileum, and malondialdehyde (MDA) level in the ileum were decreased of the DEX challenged broilers. The DEX challenge decreased microtubule-associated protein 1 light chain 3 (LC3)-II, Beclin1, and autophagy-related gene (ATG) 7 mRNA expressions, and the LC3-II/LC3-I value and increased LC3-I, cysteinyl aspartate specific proteinase (Caspase)-3 and Caspase-9 mRNA expressions in the ileum, which were improved by CGA. DEX also decreased the protein expressions of Kelch-like ECH-associated protein-1 (Keap1), Nrf2, HO-1, NADPH quinone oxidoreductase-1(NQO-1) and increased sequestosome 1 (p62) in the ileum, which were improved by CGA. Interactions occurred between DEX and CGA for the ADG, F:G ratio, villus height, crypt depth, V/C value, and SOD, CAT, GST, and HO-1 activities, MDA and PCO levels, LC3-II/LC3-I value, and expressions of LC3-I, LC3-II, Beclin1, ATG7, Caspase-3, Caspase-9, Occludin, ZO-1, Keap1, Nrf2, HO-1, NQO-1, and p62. In conclusion, CGA improved the growth performance and intestinal health of oxidatively stressed broilers by activating the autophagy-mediated Nrf2 pathway.

Effects of Chlorogenic Acid on Performance, Anticoccidial Indicators, Immunity, Antioxidant Status, and Intestinal Barrier Function in Coccidia-Infected Broilers.

The effects of chlorogenic acid (CGA) on growth performance, anticoccidial indicators (oocysts per gram of excreta, cecal lesion score, and bloody diarrhea score), immunity, antioxidant status, and intestinal barrier function in coccidia-infected broilers were investigated. A total of 240 one-day-old Arbor Acres broilers were randomly divided into four groups with six replicates of ten broilers each for 42 days. Four treatments included control diet (non-infected control, NC), control diet +Eimeria infection (infected control, IC), control diet +0.5 g/kg CGA + Eimeria infection (CGA0.5), and control diet +1 g/kg CGA + Eimeria infection (CGA1). At day 14, each broiler in IC, CGA0.5, and CGA1 groups was orally inoculated with 1 mL saline containing 4 × 105 sporulated oocysts. The results showed that the CGA1 group increased the average daily gain by 12.57% (p < 0.001) and decreased the feed/gain ratio (p = 0.010) and mortality (p = 0.030) by 13.00% and 77.76%, respectively, of broilers from 14 to 42 days compared with the IC group. The CGA1 group decreased the oocysts per gram of excreta (p < 0.001) and bloody diarrhea score (p = 0.001) compared with the IC group. The CGA0.5 and CGA1 groups increased total antioxidant capacity (p < 0.001) at day 21 and villus height (p < 0.001) in the duodenum and jejunum at day 42, and decreased the levels of interleukin 6 (IL-6) (p = 0.002), malondialdehyde (MDA) (p < 0.001), D-lactic acid (p < 0.001), and diamine oxidase (DAO) (p < 0.001) at day 21 and the levels of MDA (p < 0.001) and D-lactic acid (p = 0.003) at day 42 compared with the IC group. In the CGA1 group, villus height in the duodenum (p < 0.001) and jejunum (p = 0.017) increased at day 21 and in the ileum (p < 0.001) at day 42, and the level of DAO (p < 0.001) decreased at day 42 compared with the IC group. Broilers in the IC group had a higher IL-6 level (p = 0.048) at day 42 and lower IL-10 (p = 0.027) and immunoglobulin A (p = 0.042) levels at day 21, and IL-10 level (p = 0.017) at day 42 than those in the NC group, while no significant differences were observed among the NC, CGA0.5, and CGA1 groups. In conclusion, dietary supplementation with 1 g/kg CGA improved growth performance, immunity, antioxidant status, and intestinal barrier function in coccidia-infected broilers.

Protective effects of chlorogenic acid on the meat quality of oxidatively stressed broilers revealed by integrated metabolomics and antioxidant analysis.

Oxidation is a major cause of meat quality deterioration during broiler production, which leads to undesirable meat color and impaired water holding capacity (WHC), thereby impacting consumer appeal and satisfaction. Chlorogenic acid (CGA), a natural phenolic acid, is regarded as a potential, safer and healthier antioxidant to improve meat quality. To investigate the protective effects of CGA on the meat quality of oxidatively stressed broilers, 240 one-day-old male Cobb broiler chickens were allocated to four treatments: basal diet (control group), basal diet + dexamethasone (DEX) injection (DEX group), basal diet containing 500 mg kg-1 CGA (CGA group), and basal diet containing 500 mg kg-1 CGA + DEX injection (DEX_CGA group). Meat quality, antioxidant capacity, the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, and metabolomic profile were detected in the breast muscle of broilers. Then, correlation analysis between meat quality and antioxidant capacity, antioxidant-related genes, and metabolites was performed. The results indicated that CGA supplementation improved the growth performance and meat quality traits (pH, WHC, and meat color) and enhanced the antioxidant enzyme activity by activating the Nrf2 pathway in the breast muscle of oxidatively stressed broilers. A total of 619 metabolites were identified, among which 93 differential metabolites were found between control and DEX groups, and 65 differential metabolites were observed between DEX and DEX_CGA groups. Breast metabolic profiles were changed by DEX treatment, while CGA supplementation could normalize the metabolic changes in DEX-challenged broilers. Metabolic pathway analysis revealed that most of the differential metabolites between DEX and DEX_CGA groups were involved in pyrimidine/purine, propanoate and phenylalanine metabolism, primary bile acid biosynthesis, and lysine metabolism, which may contribute to explain the protective effects of CGA on meat quality. Moreover, according to the correlation analysis, four metabolites were identified as potential biomarkers to predict the meat quality. In conclusion, our findings demonstrate that CGA is an effective, natural and safe antioxidant to enhance the quality of meat from intensive industrial poultry production.

Interactions between Human Gut Microbiome Dynamics and Sub-Optimal Health Symptoms during Seafaring Expeditions.

During long ocean voyages, crew members are subject to complex pressures from their living and working environment, which lead to chronic diseases-like sub-optimal health status. Although the association between dysbiotic gut microbiome and chronic diseases has been broadly reported, the correlation between the sub-optimal health status and gut microbiome remains elusive. Here, the health status of 77 crew members (20-35 years old Chinese, male) during a 135-day sea expedition was evaluated using the shotgun metagenomics of stool samples and health questionnaires taken before and after the voyage. We found five core symptoms (e.g., abnormal defecation frequency, insomnia, poor sleep quality, nausea, and overeating) in 55 out of 77 crew members suffering from sub-optimal health status, and this was termed "seafaring syndrome" (SS) in this study. Significant correlation was found between the gut microbiome and SS rather than any single symptom. For example, SS was proven to be associated with individual perturbation in the gut microbiome, and the microbial dynamics between SS and non-SS samples were different during the voyage. Moreover, the microbial signature for SS was identified using the variation of 19 bacterial species and 26 gene families. Furthermore, using a Random Forest model, SS was predicted with high accuracy (84.4%, area under the concentration-time curve = 0.91) based on 28 biomarkers from pre-voyage samples, and the prediction model was further validated by another 30-day voyage cohort (accuracy = 83.3%). The findings in this study provide insights to help us discover potential predictors or even therapeutic targets for dysbiosis-related diseases. IMPORTANCE Systemic and chronic diseases are important health problems today and have been proven to be strongly associated with dysbiotic gut microbiome. Studying the association between the gut microbiome and sub-optimal health status of humans in extreme environments (such as ocean voyages) will give us a better understanding of the interactions between observable health signs and a stable versus dysbiotic gut microbiome states. In this paper, we illustrated that ocean voyages could trigger different symptoms for different crew member cohorts due to individual differences; however, the co-occurrence of high prevalence symptoms indicated widespread perturbation of the gut microbiome. By investigating the microbial signature and gut microbiome dynamics, we demonstrated that such sub-optimal health status can be predicted even before the voyage. We termed this phenomenon as "seafaring syndrome." This study not only provides the potential strategy for health management in extreme environments but also can assist the prediction of other dysbiosis-related diseases.

Investigating the growth performance, meat quality, immune function and proteomic profiles of plasmal exosomes in Lactobacillus plantarum-treated broilers with immunological stress.

Exosomes are extracellular membranous nanovesicles that carry functional molecules to mediate cell-to-cell communication. To date, whether probiotics improve the immune function of broilers by plasmal exosome cargo is unclear. In this study, 300 broilers were allocated to three treatments: control diet (CON group), control diet + dexamethasone injection (DEX group), and control diet containing 1 × 108 cfu g-1 P8 + DEX injection (P8 + DEX group). The growth performance, meat quality and immune function of plasma and jejunal mucosa were detected. Exosomes were isolated from the plasma and characterized. Then, the exosome protein profile was determined by proteomic analysis. Correlation analyses between the exosomal proteins and growth performance, meat quality, immune function were performed. Lastly, the related protein levels were verified by multiple reaction monitoring (MRM). Results showed that P8 treatment increased the growth performance, meat quality and immune function of DEX-induced broilers with immunological stress. Moreover, the average diameters, cup-shaped morphology and expressed exosomal proteins confirmed that the isolated extracellular vesicles were exosomes. A total of 784 proteins were identified in the exosomes; among which, 126 differentially expressed proteins (DEPs) were found between the DEX and CON groups and 102 DEPs were found between the P8 + DEX and DEX groups. Gene ontology analysis indicated that DEPs between the DEX and CON groups are mainly involved in the metabolic process, cellular anatomical entity, cytoplasm, etc. DEPs between the P8 + DEX and DEX groups are mainly involved in the multicellular organismal process, response to stimulus, cytoplasm, etc. Pathway analysis revealed that most of the DEPs between the DEX and CON groups participated in the ECM-receptor interaction, focal adhesion, regulation of actin cytoskeleton, etc. Most of the DEPs between the P8 + DEX and DEX groups participated in the ErbB and PPAR signaling pathways. Moreover, many DEPs were correlated with the altered parameters of growth performance, meat quality and immunity in P8-treated broilers. MRM further revealed that the upregulated FABP6 and EPCAM in the DEX group were decreased by P8 + DEX treatment, and the downregulated C1QTNF3 in the DEX group was increased by P8 + DEX treatment. In conclusion, our findings demonstrated that P8 may promote the immune function, growth performance and meat quality of broilers with immunological stress by regulating the plasma exosomal proteins, especially the proteins of FABP6, EPCAM and C1QTNF3 and the pathway of PPAR (ILK/FABP6).