Integrated transcriptomics and metabolomics study of embryonic breast muscle of Jiaji ducks.

Because number of matured muscle fibers in poultry does not increase after birth, the meat yield is mainly determined during embryogenesis. We previously indicated breast muscle grew rapidly from 18th day after hatching (E18) to E27, and almost stopped from E27 to E34 of Jiaji ducks, while the mechanism is unclear. This study utilized RNA-seq to explore the related genes of muscle development and their relationship with small molecule metabolites at E18, E27 and E34 of Jiaji ducks. Several thousand differentially expressed genes (DEGs) were detected among E18, E27 and E34. DEGs expression profiles included 8 trend maps, among which trend 1 was opposite to and trend 6 was consistent with breast muscle development trend of Jiaji ducks. Through joint analysis between trend 1 of DEGs and trend 1 of differential metabolites (DEMs), protein digestion and absorption pathway stood out. The decrease of COL8A2 gene expression will lead to the decrease of arginine content, which will inhibit the development of breast muscle in embryonic Jiaji duck. Similarly, joint analysis between trend 6 of DEGs and trend 6 of DEMs indicated the increase of GAMT gene expression will cause the increase of proline content, and then promote the development of breast muscle of Jiaji duck in embryonic period. These results will be helpful for further understanding the mechanism of muscle yields of Jiaji ducks.

Impact of dietary protein and energy levels on fatty acid profile, gut microbiome and cecal metabolome in native growing chickens.

The present study investigated the optimal concentration of dietary ME and CP for the fatty acid profile of meat, gut microbiome, and cecal metabolome in Danzhou chickens from 120 to 150 d of age. A total of seven hundred and twenty 120-d-old Danzhou female chickens, with a similar BW, were randomly allocated into 6 treatments with 6 replicates and each of 20 birds. The chickens were fed 2 levels of dietary ME (11.70 MJ/kg, 12.50 MJ/kg), and 3 levels of dietary CP (13%, 14%, and 15%). The results showed that dietary ME and CP levels didn't affect final BW, ADG, ADFI, and feed gain ratio (g: g) (P > 0.05). The serum concentrations of triglyceride, insulin, and glucose in the 12.50 MJ/kg group were the highest (P < 0.05). Dietary ME, CP levels, and their interactions affected (P < 0.05) the fatty acid content in the breast muscle, thigh muscle, and liver. The levels of C18:0, C20:0, C22:0, C22:1, C18:2, C18:3, C22:6, and SFA of the liver in the high ME group were higher than those in the low ME group (P < 0.05). The levels of C16:0, C14:1, C18:1, C22:5, SFA, MUFA and USFA in the low CP group were higher than the corresponding values in the other groups (P < 0.05). Dietary ME and CP levels altered the composition and relative abundance of microbiota in the cecum of chickens at various taxonomic levels to different extents. Significant effects of interactions were found between dietary ME and CP on the relative abundance of 10 species (P < 0.05), and among these species, 6 species belonged to the genus Bacteroides. Notably, the relative abundance of 2 probiotic species including Lactobacillus crispatus and Lactobacillus salivarius was significantly increased (P < 0.05) with increasing dietary ME level. There were 6 differential metabolites in the cecum, comprising thromboxane A2, 5,6-DHET, prostaglandin D2, 20-hydroxyeicosatetraenoic acid, 12(S)-HPETE and prostaglandin I2 significantly reduced (P < 0.05) with increasing the dietary ME level; all of them are involved in arachidonic acid metabolism. In conclusion, the present study suggested that the dietary levels of 12.50 MJ/kg ME and 14% CP enhanced meat quality in terms of fatty acid composition, and showed benefits for maintaining intestinal health via positive regulation of cecal microbiota in native growing Danzhou chickens.

Effects of mixed extract from two tropical plants on gut microbiome and metabolome in piglets.

In this study, we performed a quantitative analysis of 12 compounds derived from Piper sarmentosum extract (PSE) and guava leaf extract (GE). In addition, we investigated the effects of mixed extract (ME) of PSE and GE (1:1) on piglets' gut microbiome and metabolome. A total of 200 piglets (Duroc × Landrace × Large Yorkshire, 21-day-old) were randomly assigned into two groups with five replicates of 20 piglets/pen having the same initial body weight. Piglets were fed a basal diet supplemented with ME at 0 (T0) or 200 mg/kg (T1) for 3 weeks. The quantitation results by ultraperformance liquid chromatography linked to triple-quadrupole tandem mass spectrometry showed that vitexin 2-O-rhamnoside and pellitorine were the greatest abundant among six compounds detected in the PSE. In addition, quercetin, isoquercitrin and avicularin were found to be the richest of all detected compounds in the GE. Findings on experimental animals indicated that three differential metabolites, comprising L-alanine, sarcosine and dihydrofolic acid, in T1 compared with T0 groups, have exactly opposite levels trends in serum and faeces. Moreover, two metabolic pathways (i.e., urea cycle and glutamate metabolism) differed significantly in the serum and faeces of piglets between T0 and T1 (p < 0.05). At the same time, T1 had significantly higher relative abundances of Agathobacter and Alloprevotella than T0 at genus level (p < 0.05). Correlation analysis revealed that the genus Agathobacter correlated positively with carbamoyl phosphate (p < 0.01) and oxoglutaric acid (p < 0.05), and negatively with succinic acid (p < 0.01) and ornithine (p < 0.05). These four differential metabolites were also involved in the urea cycle and/or glutamate metabolism pathways. The results here indicated that the tested plant extract mixture represents a worthy feed additive with obvious antioxidative properties.

Transcriptomic analysis of bovine endometrial epithelial cells in response to interferon tau and hormone stimulation.

The embryonic loss during early stage of gestation is one of the major causes of infertility for domestic ruminants, causing huge economic losses to pasture. Maternal recognition of pregnancy and implantation are the crucial process for determining the successful establishment and development of pregnancy in cattle. The research on molecular mechanisms of pregnancy recognition will facilitate illustrating the complex process of pregnancy establishment and help to improve pregnancy outcomes. In this study, we performed transcriptomic analysis of primary bovine endometrial epithelial cells (BEND) with or without IFNT and hormones intervention through RNA sequencing. We eventually identified 608 differentially expressed genes (DEGs) including 409 up-regulated genes and 199 down-regulated genes in IFNT and hormones-treated group compared with control group. Gene Ontology (GO) enrichment analysis demonstrated that the majority of DEGs were implicated in immune system process, response to external stimulus, response to cytokine, regulation of response to stress. Results from KEGG analysis showed a significant enrichment of NOD-like receptor signaling pathway, antigen processing and presentation, necroptosis, oxidative phosphorylation, RIG-I-like receptor signaling pathway. Additionally, a set of promising candidate genes, including (USP18, STAT1, PSMB8, IFIH1, MX2, IFI44, DHX58, CASP8, DRAM1, CXCR4), were characterized by constructing an integrated interaction network. Specifically, the mRNA expression of HOXA11, PTGS1 and PTGS2 were remarkably suppressed by silencing DRAM1 under IFNT and hormone administration, thus speculating that DRAM1 might play a crucial role in early pregnancy by regulating endometrial function. The results of this study depicted a relatively comprehensive transcriptional profiles of BEND in response to IFNT and hormones, which contributes to a better understanding of gene interaction network and underlying regulatory mechanisms in endometrium of ruminants during early pregnancy.

Antagonistic Activity of Oroxylin A against Fusarium graminearum and Its Inhibitory Effect on Zearalenone Production.

Fusarium graminearum produces zearalenone (ZEA), a mycotoxin that is widely found in food and feed products and is toxic to humans and livestock. Piper sarmentosum extract (PSE) inhibits F. graminearum, and Oroxylin A appears to be a major antifungal compound in PSE. The aim of this study is to quantify the Oroxylin A content in PSE using UPLC-QTOF-MS/MS, and to investigate the antagonistic activity of Oroxylin A against F. graminearum and its inhibitory effect on ZEA production. The results indicate that Oroxylin A inhibits both fungal growth and ZEA production in a dose-dependent manner. Oroxylin A treatment downregulated the mRNA expression of zearalenone biosynthesis protein 1 (ZEB1) and zearalenone biosynthesis protein 2 (ZEB2). The metabolomics analysis of F. graminearum mycelia indicated that the level of ribose 5-phosphate (R5P) deceased (p < 0.05) after Oroxylin A treatment (64-128 ng/mL). Moreover, as the Oroxylin A treatment content increased from 64 to 128 ng/mL, the levels of cis-aconitate (p < 0.05) and fumarate (p < 0.01) were upregulated successively. A correlation analysis further showed that the decreased R5P level was positively correlated with ZEB1 and ZEB2 expression, while the increased cis-aconitate and fumarate levels were negatively correlated with ZEB1 and ZEB2 expression. These findings demonstrate the potential of Oroxylin A as a natural agent to control toxigenic fungi and their mycotoxin.

The Influence of Piper sarmentosum Extract on Growth Performance, Intestinal Barrier Function, and Metabolism of Growing Chickens.

In the poultry industry, there is an urgent need to evaluate and introduce natural, effective, and safe alternatives for synthetic antibiotics, which have been banned in most countries. The present study aimed to investigate the effects of dietary supplementation with Piper sarmentosum extract (PSE) on the growth performance, intestinal barrier function, and metabolism of growing chickens. A total of 400 seven-day-old female chicks were randomly assigned to four dietary treatments, each of which consisted of five replicates and twenty birds each. The four experimental treatments were fed a basal diet containing 0, 100, 200, and 300 mg PSE/kg (BC, PSE1, PSE2, and PSE3 groups), respectively. The experiment lasted for 28 days. The results showed that dietary supplementation with PSE had no significant effects on the final body weight, average daily gain (ADG), average daily feed intake (ADFI), and the ratio of ADFI to ADG (F/G) (p > 0.05). Compared with the BC group, dietary supplementation with 200-300 mg/kg PSE increased the villus height in the jejunum and ileum of chickens (p < 0.05). The PSE-treated groups significantly increased the mRNA expression of Occludin, ZO-1, and Claudin-1 in the ileal mucosa of chickens (p < 0.05). In addition, a significant decrease in ileal TNF-α and IL-8 mRNA expression (p < 0.05) and a significant increase in IL-22 (p < 0.05) were observed in the PSE2 treatment compared to the BC group. Additionally, three gut metabolites (i.e., citrate, isocitrate, and spermine) showed significant differences among treatments (p < 0.05) and were involved in the tricarboxylic acid (TCA) cycle, the transfer of acetyl groups into mitochondria, and spermidine and spermine biosynthesis, respectively. In conclusion, the findings obtained here indicate that supplemental PSE can enhance the anti-inflammatory capacity and intestinal mucosal barrier function of chickens.

Antifungal activity and metabolomics analysis of Piper sarmentosum extracts against Fusarium graminearum.

AIMS: Fusarium graminearum is a toxic fungus that affects food and feed crops. Piper sarmentosum extract (PSE) is a potential source of anti-mildew natural products for the food and feed industry due to its various pharmacological properties. In this study, we evaluated the antifungal activity and untargeted metabolomics analysis of PSE against F. graminearum. METHODS AND RESULTS: Antifungal activity was evaluated using the mycelium growth rate method. Untargeted metabolomics analysis of PSE was performed using ultra high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The results showed that PSE (1 and 2 mg ml-1) possesses inhibitory activity against F. graminearum, and a total of 17 compounds that including 8 alkaloids, 3 phenols, 3 lipids, and 3 organic acids might be the antifungal markers in PSE. Metabolomics analysis further revealed that PSE could significantly increase the levels of guanosine, guanine, adenosine, and L-isoleucine in fungi, which are related to purine and L-isoleucine metabolic pathways. CONCLUSIONS: PSE is a promising anti-mildew agent that inhibits the growth of F. graminearum in food and feed. PSE (1 and 2 mg ml-1) may exert antifungal properties by inhibiting fungal purine nucleotide synthesis and enhancing the level of L-isoleucine compared with the control groups.

Effects of blended microbial feed additives on performance, meat quality, gut microbiota and metabolism of broilers.

The present study investigated the effects of blend microbial feed additive (BMFA) in diet on performance, meat quality, gut microbiota and metabolism of broilers. In this study 240 seventy-day-old female Wenchang broilers were randomly allocated into four groups with five replicates of 12 broilers each. Broilers in the control group was fed only basal diet (S0), and the other three groups were fed the same basal diet supplemented with 0.2% (S1), 0.4% (S2), or 0.6% (S3) of BMFA, respectively. The trial continued for 54 days. The results showed that broilers in S2 and S3 had lower average daily feed intake (ADFI) compared with S0 and S1 (P < 0.05). However, diet supplementation with BMFA had no significantly influence on the average daily gain (ADG) and the ratio of ADFI to ADG (F/G) (P > 0.05). The highest thigh muscle percentage was observed in S2 (P < 0.05) among all groups. Diet supplementation with BMFA reduced the shear force in both breast and thigh muscles (P < 0.05) of broilers. An increase (P < 0.05) in the total unsaturated fatty acid (USFA), monounsaturated fatty acids (MUFA), and ratio of unsaturated fatty acids to saturated fatty acid (USFA/SFA) in breast muscles was observed in S3 compared with S0. It was found that the S3 had a relatively higher abundance of Lactobacillus (P < 0.001), as well as a lower abundance of the Bacteroides, Rikenellaceae RC9 gut group, Olsenella, Prevotellaceae UCG-001 and Prevotella (P < 0.05) than the S0. Correlation analysis indicated that a total of 17 differential metabolites between the S3 and S0 were significantly correlated with the 7 differential genera microflora. Overall, diet supplementation with 0.6% of BMFA can significantly improve the meat quality of broilers by decreasing the concentration of SFA and enhancing the levels of the total USFA, MUFA and USFA/SFA in breast muscles. Those findings were tightly bound to the higher proportion of Lactobacillus genus in the intestinal tract of broilers influenced by BMFA.

Integrated analysis of lncRNA and gene expression in longissimus dorsi muscle at two developmental stages of Hainan black goats.

It is deemed that meat quality of kids' is better than that of adults' for Hainan black goat. Generally, meat quality is affected by many indicators, such as intramuscular fat (IMF) content, muscle fiber diameter and shear force. It is indicated that long non-coding RNAs (lncRNAs) play essential roles in meat quality of goats. However, it is unclear whether and how lncRNAs and genes play their roles in meat quality of Hainan Black goats. Here, we firstly compared the meat quality between two-month-old kids (kids) and adult goats (adults). Then, the lncRNA-seq and RNA-seq data were integrated and analyzed to explore the potential functions of lncRNAs and genes. The results showed that adults' IMF content and muscle fiber diameter were extremely significantly higher than that of kids (P<0.01). For the sequenced data, average 84,970,398, and 83,691,250 clean reads were obtained respectively for Kids and adults, among which ~96% were mapped to the reference genome of goats. Through analyzing, 18,242 goat annotated genes, 1,429 goat annotated lncRNAs and 2,967 novel lncRNAs were obtained. Analysis of differential expression genes (DEGs) and lncRNAs (DELs) showed that 328 DEGs and 98 DELs existed between kids and adults. Furthermore, functional enrichment analysis revealed that a number of DEGs and DELs were mainly associated with IMF. Primarily, DGAT2 expressed higher in adults than that in kids and CPT1A expressed higher in kids than that in adults. Both of them were overlapped by DEGs and targets of DELs, suggesting the two DEGs and the DELs targeted by the two DEGs might be the potential regulators of goat IMF deposition. Taken together, our results provide basic support for further understanding the function and mechanism of lncRNAs and genes in meat quality of Hainan black goats.

Effects of Vine Tea Extract on Meat Quality, Gut Microbiota and Metabolome of Wenchang Broiler.

This study investigates the effects of vine tea (Ampelopsis grossedentata) extract (AGE) on meat quality, gut microbiota and cecal content metabolites of Wenchang broilers. A total of 240 female Wenchang broilers aged 70 days were randomly allocated into four groups with five replicates of twelve broilers each. Broilers were fed a corn-soybean basal diet supplemented with AGE at 0 (T1), 0.2% (T2), 0.4% (T3) and 0.6% (T4) until 124 days of age. The whole feeding trial lasted 54 days. Results suggest that the content of total triglycerides and low-density lipoprotein cholesterol in serum of broilers are linearly reduced with dietary AGE supplementation (p < 0.05). The T3 and T4 groups had higher (p < 0.05) a* value in thigh and breast muscles than the T1 group. Additionally, the dietary supplementation of AGE decreased the shear force and drip loss of both thigh and breast muscles linearly (p < 0.05). Compared with the T1 group, AGE supplementation increased the levels of inosine monophosphate (IMP) significantly (p < 0.05) in both the thigh and breast muscles. Furthermore, an increase (p < 0.05) in the total unsaturated fatty acid (USFA), polyunsaturated fatty acids (PUFA) and the ratio of unsaturated fatty acids to saturated fatty acid (USFA: SFA) in both the thigh and breast muscles in the T3 group was observed. Higher abundance of Bacteroidota (p < 0.05) and lower abundance of Firmicutes (p < 0.05) were observed in the T3 group. The abundance of Faecalibacterium was significantly decreased (p < 0.05) in the T3 group compared with the T1 group. Cholesterol sulfate and p-cresol sulfate were identified as differential metabolites between the T1 and T3 groups. It suggested that 0.4% of AGE supplementation significantly downregulated the levels of p-cresol sulfate and cholesterol sulfate (p < 0.05) and the hepatic 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) activity compared with the control. Our present study demonstrates that dietary supplementation with AGE can improve the quality and flavor by increasing the IMP and PUFA content in the muscle of Wenchang broilers. Furthermore, dietary AGE supplementation with 0.4% can regulate the cholesterol metabolism of Wenchang broilers.

Contribution of the mutation rs8193069 in TLR4 to mastitis resistance and performance in Holstein cows in southern China.

Bovine mastitis has become increasingly important issues for farmers and consumers, leading to large economic losses in the dairy industry worldwide. Because treatment of mastitis is difficult and costly, improved mastitis resistance through selective breeding would be advantageous. The toll-like receptor 4 (TLR4) is an important player in recognising pathogens and activating immune responses. However, its roles in mastitis occurrence and the underlying molecular mechanisms are unclear. In this study, a single nucleotide polymorphism, rs8193069 (T â†’ C) in TLR4 gene was detected in a Holstein cow resource population in southern China. Association analysis with 5-year production traits, haematology, and biochemistry parameters revealed that individuals with genotype CC had significantly lower somatic cell counts (SCC), lower fat percentage, but higher 305-day milk (p < 0.05) and total milk yield (p < 0.01). Both genotypes CC and CT had lower lymphocyte counts (#LYMPH) (p < 0.01) and basophil counts (#BASO) (p < 0.05) than TT. Genotype CC had a less level of triglyceride (p < 0.01) and creatine kinase (p < 0.05) than CT. Further analysis based on the production data revealed significant positive correlations between SCC and #LYMPH. Analysis of TLR4 protein structure and properties suggested that the missense mutation on the 674th amino acid from Thr to Ile reduced the flexibility and hydrophilicity of TIR domain, implying a weakened binding ability of TLR4 to its adaptors. In conclusion, allele C of rs8193069 was the major allele in Holstein cows that indicated a greater genetic potential to mastitis resistance and milk yields, probably via the LPS-TLR4 inflammatory signalling. This study offers a marker to improve mastitis resistance in the dairy cow population in southern China.

Dietary Supplementation With Didancao (Elephantopus scaber L.) Improves Meat Quality and Intestinal Development in Jiaji Ducks.

Didancao (Elephantopus scaber L.) has been used as a traditional herbal medicine and has exhibited a beneficial role in animal health. This study aimed to investigate the effects of dietary supplementation with E. scaber on growth performance, meat quality, intestinal morphology, and microbiota composition in ducks. A total of 480 Jiaji ducks (42 days old, male:female ratio = 1:1) were randomly assigned to one of four treatments. There were six replicates per treatment, with 20 ducks per replicate. The ducks in the control group (Con) were fed a basal diet; the three experimental groups were fed a basal diet supplementation with 30 (T1), 80 (T2), and 130 mg/kg (T3) of E. scaber. After a 48-day period of supplementation, growth performance, meat quality, intestinal morphology, and microbiota composition were evaluated. The results showed that no differences were observed in the final body weight, average daily feed intake, and average daily gain among the four groups. Compared with that in the Con group, the feed conversion in the T1 and T2 groups was increased significantly; the T2 group was shown to decrease the concentration of alanine aminotransferase in serum; the T3 group was lower than the Con group in the concentration of aspartate aminotransferase and was higher than the Con group in the concentration of high-density lipoprotein-cholesterol. The highest concentration of creatinine was observed in the T1 group. The T2 group was higher than the Con group in the contents of Phe, Ala, Gly, Glu, Arg, Lys, Tyr, Leu, Ser, Thr, Asp, and total amino acids in the breast muscle. Moreover, the T2 group was higher than the Con group in the contents of meat C18:2n-6 and polyunsaturated fatty acid. The concentration of inosinic acid in the T1, T2, and T3 groups was significantly higher than that in the Con group. However, the Con group was higher than the T2 or T3 group in the Zn content. The T2 group was lower than the Con group in the jejunal crypt depth. The T3 group was higher than the Con group in the ileal villus height and the ratio of villus height to crypt depth. In addition, the T3 group had a trend to significantly increase the abundance of Fusobacteria. Compared with the Con group, the T1 and T2 groups displayed a higher abundance of Subdoligranulum. Collectively, dietary supplementation with 80 mg/kg of E. scaber improves meat quality and intestinal development in ducks.

Effects of Guava (Psidium guajava L.) Leaf Extract on the Metabolomics of Serum and Feces in Weaned Piglets Challenged by Escherichia coli.

The effects of dietary supplementation with guava leaf extracts (GE) on intestinal barrier function and serum and fecal metabolome in weaned piglets challenged by enterotoxigenic Escherichia coli (ETEC) were investigated. In total, 50 weaned piglets (Duroc × Yorkshire × Landrace) from 25 pens (two piglets per pen) were randomly divided into five groups: BC (blank control), NC (negative control), S50 (supplemented with 50 mg kg-1 diet GE), S100 (100 mg kg-1 diet GE), and S200 (200 mg kg-1 diet GE), respectively. On day 4, all groups (except BC) were orally challenged with enterotoxigenic ETEC at a dose of 1.0 × 109 colony-forming units (CFUs). After treatment for 28 days, intestinal barrier function and parallel serum and fecal metabolomics analysis were carried out. Results suggested that dietary supplementation with GE (50-200 mg kg-1) increased protein expression of intestinal tight junction proteins (ZO-1, occludin, claudin-1) (p < 0.05) and Na+/H+ exchanger 3 (NHE3) (p < 0.05). Moreover, dietary supplementation with GE (50-200 mg kg-1) increased the level of tetrahydrofolic acid (THF) and reversed the higher level of nicotinamide-adenine dinucleotide phosphate (NADP) induced by ETEC in serum compared with the NC group (p < 0.05), and enhanced the antioxidant capacity of piglets. In addition, dietary addition with GE (100 mg kg-1) reversed the lower level of L-pipecolic acid induced by ETEC in feces compared with the NC group (p < 0.05) and decreased the oxidative stress of piglets. Collectively, dietary supplementation with GE exhibited a positive effect on improving intestinal barrier function. It can reprogram energy metabolism through similar or dissimilar metabolic pathways and finally enhance the antioxidant ability of piglets challenged by ETEC.

Chemical composition and anti-inflammatory activity of n-butanol extract of Piper sarmentosum Roxb. In the intestinal porcine epithelial cells (IPEC-J2).

ETHNOPHARMACOLOGICAL RELEVANCE: Piper sarmentosum Roxb. (PS) is a terrestrial herb primarily distributed in tropical and subtropical regions of Asia. It is widely used in folk medicine in certain countries of Southeast Asia for the treatment of fever, toothache, coughing and pleurisy, which showed the anti-inflammatory activity of PS. AIM OF THE STUDY: This study aimed to investigate the chemical constituents and the molecular mechanism and related metabolic pathway by which n-butanol extract of PS (PSE-NB) exerts its anti-inflammatory effects. MATERIALS AND METHODS: Chemical constituents of PSE-NB was analyzed using ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technique. Anti-inflammatory effects of PSE-NB were investigated in lipopolysaccharide (LPS)-induced IPEC-J2 cells. RESULTS: In total, 218 compounds, including 94 alkaloids and 26 phenolics were tentatively identified, which indicating alkaloids and phenolics were the main constituents of PSE-NB. In addition, the current cell experiment in vitro showed that PSE-NB (10-500 µg/mL) pre-treatment before LPS stimulation significantly decreased mRNA expression of IL-1ß, IL-6 and TNF-α in IPEC-J2 cells compared with LPS treatment (p < 0.05). PSE-NB improved mRNA expression of tight junction proteins (ZO-1 and Occludin) and NHE3, which were reduced by LPS stimulation (p < 0.05). Moreover, PSE-NB (10 µg/mL) alleviated LPS-induced protein expression of p65 and p-p65 (p < 0.05), and reduced p65 translocation into the nucleus induced by LPS. At the same time, metabolic pathway analysis indicated that PSE-NB exerts anti-inflammatory effects mainly via augmentation of methionine metabolism in IPEC-J2 cells. CONCLUSIONS: Taken together, the results suggested that alkaloids and phenolics were the main constituents in PSE-NB. PSE-NB might attenuate LPS-induced inflammatory responses in IPEC-J2 cells by regulating NF-κB signaling pathway and intracellular metabolic pattern.

Using Extract From the Stems and Leaves of Yizhi (Alpiniae oxyphyllae) as Feed Additive Increases Meat Quality and Intestinal Health in Ducks.

Yizhi (Alpiniae Oxyphyllae, A. oxyphylla) has been widely used as an important traditional Chinese medicinal herb for centuries. Existing studies have shown that A. oxyphylla has numerous benefits in human and animal health. We hypothesized that extract from the stems and leaves of A. oxyphylla (AOE) as a feed additive may have positive effects on animal health and products. Thus, this study was conducted to evaluate the effects of AOE as a feed additive on growth performance, serum biochemical parameters, intestinal morphology, microbial composition, and meat quality in Jiaji ducks. A total of 240 Jiaji ducks of 42 days old (1675.8 ± 44.2 g, male: female ratio = 1:1) were blocked based on body weight and randomly allocated into four dietary treatments with three replicates that each had 20 duck individuals. The dietary treatments included: basal diet, control group (CK); basal diet supplementation with 30 mg/kg (Y1), 80 mg/kg (Y2), and 130 mg/kg (Y3) AOE, respectively, and lasted for 49 days. The results showed that average daily feed intake from day 42 to day 60 was decreased with the increasing level of AOE (P < 0.05). Compared with the CK group, the groups with AOE supplementation decreased serum LDL-C level (P < 0.05), the addition of 30 mg/kg AOE increased total amino acids, essential amino acids, branched-chain amino acids, nonessential amino acids, and umami taste amino acids (P < 0.05), but decreased selenium and zinc concentrations in breast muscle (P < 0.05). In addition, the supplementation of 30 or 130 mg/kg AOE significantly increased jejunal villus height (P < 0.05) and tended to increase the ratio of villus height to crypt depth in the jejunum (P = 0.092) compared to the CK group. Moreover, the addition of 30 mg/kg AOE showed a higher abundance of genus unclassified Bacteroidales and genus unclassified Ruminococcaceae than the CK group (P < 0.05). Therefore, dietary supplementation with 30 mg/kg AOE increased meat nutrition profile and flavor through promoting amino acid contents in breast muscle, as well as maintained intestine integrity and modulated the microbial composition. In conclusion, AOE as an antibiotic alternative displayed potential in maintaining intestinal health and improving meat quality.

Chemical composition and protective effect of guava (Psidium guajava L.) leaf extract on piglet intestines.

BACKGROUND: Dietary intervention is an important approach to improve intestinal function of weaned piglets. Phytogenic and herbal products have received increasing attention as in-feed antibiotic alternatives. This study investigated the chemical composition of guava leaf extract (GE) by ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Meanwhile, we investigated the effects of dietary supplementation with GE on diarrhea in relation to immune responses and intestinal health in weaned piglets challenged by enterotoxigenic Escherichia coli (ETEC). RESULTS: In total, 323 characterized compounds, which including 91 phenolic compounds and 232 other compounds were identified. Animal experiment results showed that the supplementation of 50-200 mg kg-1 of GE in the diet could reduce diarrhea incidence, increase activities of superoxide dismutase, glutathione peroxidase and total anti-oxidant capacity in the serum (P < 0.05), decrease the levels of interleukin 1ß, interleukin 6 and tumor necrosis factor α in the serum or jejunum mucosa (P < 0.05), and increase villus height and villus height to crypt depth ratio (P < 0.05) in the jejuna of piglets challenged by oral ETEC compared with negative control group (NC). Meanwhile, diet supplementation with 50-200 mg kg-1 GE reduced the levels of D-lactate, endothelin-1 and diamine oxidase in the serum, and increased the expression of zonula occludens-1, Claudin-1, Occludin and Na+ /H+ exchanger 3 (P < 0.05) in the jejuna mucosa of piglets challenged by ETEC compared with the NC. CONCLUSIONS: These results suggested that GE could attenuate diarrhea and improve intestinal barrier function of piglets challenged by ETEC. © 2020 Society of Chemical Industry.

Identification of reliable reference genes for expression studies in maternal reproductive tissues and foetal tissues of pregnant cows.

The relationship between the conceptus and the maternal uterine environment is crucial for the successful establishment and maintenance of pregnancy in cattle. Gene expression analysis of the conceptus and maternal reproductive tissues is a favourable method to assess the embryonic maternal interaction. The reliability of the commonly used method reverse transcription-quantitative polymerase chain reaction (RT-qPCR) depends on proper normalization to stable reference genes (RGs). The objective of this study was to determine the expression stability of 10 potential RGs in maternal reproductive tissues and foetal tissues, and to analyse the effect of RG selection on the calculation of the relative expression of target genes. The expression stability of 10 potential RGs was analysed in eight different tissues from three pregnant dairy cows. Three programs-GeNorm, NormFinder and Bestkeeper-were used to identify the best RGs. According to all three programs, the most stable RG was CNOT11, whereas the least stable RGs were GAPDH and HPRT1. GeNorm analysis showed that a combination of five RGs (SDHA, PPIA, CNOT11, RPS9 and RPL19) was necessary for appropriate data normalization. However, NormFinder analysis indicated that the combination of CNOT11 and PPIA was the most suitable. When target genes were normalized to these RGs, the relative expression of the Radical S-adenosyl methionine domain containing 2 gene was not affected by the choice of RGs, whereas a large difference was observed in the expression profile of the Nuclear erythroid2-related factor 2 gene between the most stable and least stable RGs. The results indicate that careful selection of RGs is crucial under different conditions, especially for target genes with relatively small fold changes. Furthermore, the results provide useful information for the selection of RGs for evaluating genes affecting bovine reproduction.

Effects of Piper sarmentosum extract supplementation on growth performances and rumen fermentation and microflora characteristics in goats.

This study was carried out to investigate the effect of diet Piper sarmentosum extract (PSE) on the growth performance, antioxidant properties, rumen fermentation and microflora in goats. Forty Hainan black goats with similar body weight were divided into four groups with supplementation of PSE in the concentrate at 0, 300, 600 and 1,200 mg/kg, respectively, and fed for 56 days. Results showed that average daily gain (ADG) was higher and feed intake/body gain (F/G) was lower in goats fed with PSE at 300 mg/kg (p < .05). The activities of glutathione peroxidase (GSH-Px) and total antioxidant capacity (T-AOC) in the serum of goats differed among treatments and were greatest linearly when PSE was added at 1,200 mg/kg (p < .05). The level of malondialdehyde (MDA) in the serum of goats differed among treatments and was lowest linearly when PSE was added at 1,200 mg/kg (p < .05). The level of protozoal protein in the rumen of goats differed among treatments and was lowest linearly when PSE was added at 1,200 mg/kg (p < .05). The concentrations of ruminal acetic acid and valeric acid and the ratio of acetate to propionate were reduced with PSE supplementation (p < .05). Protozoa, fungi, Ruminococcus flavefaciens and Fibrobacter succinogenes contents differed among treatments and were lowest linearly when PSE was added at 1,200 mg/kg (p < .05). Thus, supplementation of PSE at 300-1,200 mg/kg to goat concentrate is recommended for improving antioxidative ability and rumen efficiency and reducing protozoal content of goat.

Landscape of alternative splicing in Capra_hircus.

Alternative splicing (AS) is a fundamental regulatory process in all higher eukaryotes. However, AS landscapes for a number of animals, including goats, have not been explored to date. Here, we sequenced 60 samples representing 5 tissues from 4 developmental stages in triplicate using RNA-seq to elucidate the goat AS landscape. In total, 14,521 genes underwent AS (AS genes), accounting for 85.53% of intron-containing genes (16,697). Among these AS genes, 6,342 were differentially expressed in different tissues. Of the AS events identified, retained introns were most prevalent (37.04% of total AS events). Functional enrichment analysis of differential and specific AS genes indicated goat AS mainly involved in organ function and development. Particularly, AS genes identified in leg muscle were associated with the "regulation of skeletal muscle tissue development" GO term. Given genes were associated with this term, four of which (NRG4, IP6K3, AMPD1, and DYSF) might play crucial roles in skeletal muscle development. Further investigation indicated these five genes, harbored 13 ASs, spliced exclusively in leg muscle, likely played a role in goat leg muscle development. These results provide novel insights into goat AS landscapes and a valuable resource for investigation of goat transcriptome complexity and gene regulation.

Anticoccidial effects of areca nut (Areca catechu L.) extract on broiler chicks experimentally infected with Eimeria tenella.

To study the anticoccidial effects of areca nut extract (ANE) on coccidiosis, 270 one-day old Wenchang broiler chicks were divided into six equal groups, each with three replicate cages (n = 15 per cage). The six groups were the blank control group (BC), negative control group (NC), positive control group (PC), and three ANE-treated groups. The birds in the three control groups (BC, NC and PC) were fed a basal diet without ANE supplementation. The birds in the three ANE-treated groups were fed a basal diet supplied with ANE at 100 (T1), 200 (T2), or 300 (T3) mg/kg feed. At 15 days of age, the birds in the NC, PC and the three ANE groups were challenged orally with 1 × 105Eimeria tenella oocysts per chick. At 48 h after oocysts inoculation, the birds in group PC were supplied diclazuril with drinking water for 5 days. The results showed that ANE and diclazuril significantly improved feed intake and body weight gain (P < 0.05) relative to the NC group. Both ANE and diclazuril significantly (P < 0.05) reduced OPG on day 4-9 post-inoculation (p.i.) relative to the NC group. Coccidial infection damaged the integrity of the cecal mucosa and thickened cecal tunica muscularis. ANE and diclazuril mitigated the mucosal damage caused by coccidial infection. Diet ANE supplementation reduced the cecal lesion scores compared to the NC group (P < 0.05). ANE and diclazuril increased nitric oxide (NO) levels at 3 days p.i., but reduced NO levels at 6 days p.i. (P < 0.05) compared to the NC group. Diet ANE supplementation increased the concentration of interleukin 2 (IL-2) in infected chicken relative to the NC group. The current results showed the anticoccidial properties, and beneficial effect on intestinal mucosa damage of ANE in broiler chicks challenged with coccidiosis.