Dietary silymarin improves performance by altering hepatic lipid metabolism and cecal microbiota function and its metabolites in late laying hens.

BACKGROUND: Liver lipid dysregulation is one of the major factors in the decline of production performance in late-stage laying hens. Silymarin (SIL), a natural flavonolignan extracted from milk thistle, is known for its hepatoprotective and lipid-lowering properties in humans. This study evaluates whether SIL can provide similar benefits to late-stage laying hens. A total of 480 68-week-old Lohmann Pink laying hens were randomly assigned into 5 groups, each group consisting of 6 replicates with 16 hens each. The birds received a basal diet either without silymarin (control) or supplemented with silymarin at concentrations of 250, 500, 750, or 1,000 mg/kg (SIL250, SIL500, SIL750, SIL1000) over a 12-week period. RESULTS: The CON group exhibited a significant decline in laying rates from weeks 9 to 12 compared to the initial 4 weeks (P = 0.042), while SIL supplementation maintained consistent laying rates throughout the study (P > 0.05). Notably, the SIL500 and SIL750 groups showed higher average egg weight than the CON group during weeks 5 to 8 (P = 0.049). The SIL750 group had a significantly higher average daily feed intake across the study period (P < 0.05), and the SIL500 group saw a marked decrease in the feed-to-egg ratio from weeks 5 to 8 (P = 0.003). Furthermore, the SIL500 group demonstrated significant reductions in serum ALT and AST levels (P < 0.05) and a significant decrease in serum triglycerides and total cholesterol at week 12 with increasing doses of SIL (P < 0.05). SIL also positively influenced liver enzyme expression (FASN, ACC, Apo-VLDL II, FXR, and CYP7A1; P < 0.05) and altered the cecal microbiota composition, enhancing species linked to secondary bile acid synthesis. Targeted metabolomics identified 9 metabolites predominantly involved in thiamin metabolism that were significantly different in the SIL groups (P < 0.05). CONCLUSIONS: Our study demonstrated that dietary SIL supplementation could ameliorate egg production rate in late stage laying hens, mechanistically, this effect was via improving hepatic lipid metabolism and cecal microbiota function to achieve. Revealed the potentially of SIL as a feed supplementation to regulate hepatic lipid metabolism dysregulation. Overall, dietary 500 mg/kg SIL had the best effects.

Effects of dietary water-soluble extract of rosemary supplementation on growth performance and intestinal health of broilers infected with Eimeria tenella.

This study was conducted to explore the effect of dietary supplementation of water-soluble extract of rosemary (WER) on growth performance and intestinal health of broilers infected with Eimeria tenella (E. tenella), and evaluate the anticoccidial activity of WER. 360 1-d-old Chinese indigenous male yellow-feathered broiler chickens were randomly allocated to six groups: blank control (BC) group and infected control (IC) group received a basal diet; positive control (PC) group, received a basal diet supplemented with 200 mg/kg diclazuril; WER100, WER200, and WER300 groups received a basal diet containing 100, 200, and 300 mg/kg WER, respectively. On day 21, all birds in the infected groups (IC, PC, WER100, WER200, and WER300) were orally gavaged with 1 mL phosphate-buffered saline (PBS) of 8 × 104 sporulated oocysts of E. tenella, and birds in the BC group were administrated an aliquot of PBS dilution. The results showed that dietary supplementation of 200 mg/kg WER increased the average daily gain of broilers compared to the IC group from days 22 to 29 (P < 0.001). The anticoccidial index values of 100, 200, and 300 mg/kg WER were 137.49, 157.41, and 144.22, respectively, which indicated that WER exhibited moderate anticoccidial activity. Compared to the IC group, the groups supplemented with WER (100, 200, and 300 mg/kg) significantly lowered fecal oocyst output (P < 0.001) and cecal coccidia oocysts, alleviated intestinal damage and maintained the integrity of intestinal epithelium. Dietary supplementation with WER significantly improved antioxidant capacity, elevated the levels of secretory immunoglobulin A, and diminished inflammation within the cecum, particularly at a dosage of 200 mg/kg. The results of this study indicated that dietary supplementation with 200 mg/kg WER could improve broiler growth performance and alleviate intestinal damage caused by coccidiosis.

Avian coccidiosis, a prevalent parasitic disease caused by Eimeria protozoa, leads to significant economic losses in the global poultry industry. Currently, the control of coccidiosis in chickens primarily relies on chemical and ionophore anticoccidials. However, the long-term use of these compounds has resulted in the development of drug-resistant strains, presenting a critical challenge. Additionally, the toxic and side effects of ionophore anticoccidials have become increasingly apparent. Thus, there is an urgent need to find economical and environmentally friendly measures to control coccidiosis in chickens. In this study, we established a model of Eimeria tenella infection in broilers to explore whether the water-soluble extract of rosemary (WER) could serve as an alternative method for controlling avian coccidiosis. Our results showed that dietary supplementation with WER (100, 200, and 300 mg/kg) had a beneficial anticoccidial effect, alleviating intestinal damage caused by coccidiosis by enhancing the intestinal antioxidant defense and activating the immune function of the infected broilers. Specifically, dietary supplementation with 200 mg/kg WER emerged as a promising strategy for controlling avian coccidiosis in the poultry industry.

Effect of Zinc Amino Acid Complexes on Growth Performance, Tissue Zinc Concentration, and Muscle Development of Broilers.

The present study aimed to evaluate the effects of zinc amino acid complexes on growth performance, tissue zinc concentration, and muscle development in broilers. A total of 504 day-old male arbor acres broilers were randomly divided into seven treatments (fed with a basal diet or a basal diet supplemented with 120 mg kg-1 Zn as ZnSO4, 30, 60, 90 or 120 mg kg-1 Zn as ZnN, or 30 mg kg-1 Zn as ZnA separately). Each group had six replicates, with 12 birds per replicate. The results showed that the addition of 60 mg kg-1 ZnN significantly increased (P < 0.05) the average daily gain (ADG) and breast muscle percentage of broilers. Zinc concentration of ZnN and ZnA added groups were higher than (P < 0.05) that in the Zn sulfate group under the same addition dose. Except for the 30 mg kg-1 ZnN group, the muscle fiber diameter and cross-sectional area (CSA) were significantly increased (P < 0.05) in the ZnN addition groups. Compared with the basal diet group, adding ZnN significantly increased (P < 0.05) the expression of MTOR, MYOD, and MYOG at day 21 and decreased (P < 0.05) the expression of Atrogin-1. The expression levels of AKT, MTOR, P70S6K, and MYOD were increased at day 42, while the expression levels of MuRF1 and Atrogin-1 were decreased. Adhesion, backbone regulation of actin, MAPK, mTOR, and AMPK were significantly enriched as indicated by KEGG pathway enrichment analysis. In conclusion, zinc amino acid complexes could improve growth performance, tissue zinc concentration, and regulate breast muscle development.

Clec7a drives gut fungus-mediated host lipid deposition.

BACKGROUND: Compared to that of bacteria, the role of gut fungi in obesity development remains unknown. RESULTS: Here, alterations in gut fungal biodiversity and composition were confirmed in obese pig models and high-fat diet (HFD)-fed mice. Antifungal drugs improved diet-induced obesity, while fungal reconstruction by cohousing or fecal microbiota transplantation maintained the obese phenotype in HFD-fed mice. Fungal profiling identified 5 fungal species associated with obesity. Specifically, Ascomycota_sp. and Microascaceae_sp. were reduced in obese mice and negatively correlated with fat content. Oral supplementation with fungi was sufficient to prevent and treat diet-induced obesity. Clec7a, which is involved in fungal recognition, was highly expressed in HFD-fed mice. The Clec7a agonist accelerated diet-induced obesity, while Clec7a deficieny in mice resulted in resistance to diet-induced obesity and blocked the anti-obese effect of antifungal drugs and fungi. CONCLUSIONS: Taken together, these results indicate that gut fungi/Clec7a signaling is involved in diet-induced obesity and may have therapeutic implications as a biomarker for metabolic dysregulation in humans. Video Abstract.

Early flora colonization affects intestinal immunoglobulin G uptake in piglets, which may be mediated by NF-κB-FcRn pathway.

Introduction: The passive immunity of newborn piglets is mainly derived from immunoglobulin G (IgG) in breast milk, and the incomplete transfer of passive immune is considered to be an important cause of piglet death. This study was conducted to investigate the effect of early intestinal flora colonization on IgG uptake and its possible mechanism. Methods: The newborn piglets and IPEC-J2 cells were used to investigate the possible factors and regulatory mechanisms affecting intestinal IgG uptake. In vivo, all 40 piglets were euthanized on postnatal d 0, 1, 3, and 7, with 10 piglets per time. The blood sample, gastric contents, jejunal contents and mucosa were collected for analysis. In vitro, IPEC-J2 cells transwell culture system was used to establish the IgG transporter model to explore the specific regulatory mechanism of IgG transport. Results: Our results demonstrated that the intestinal IgG uptake was positively correlated with the expression of Neonatal Fc receptor (FcRn). With the increase of age, the intestinal flora of newborn piglets was gradually enriched. The function of intestinal genes also changes with the colonization of intestinal flora. We found that the expression trend of TLR2, TLR4 and NF-κB (P65) in intestine was consistent with that of FcRn. Furthermore, the in vitro results demonstrate that the NF-κB signaling pathway is involved in regulating FcRn-mediated IgG transmembrane transport. Discussion: Early flora colonization affects intestinal IgG uptake in piglets, which may be mediated by NF-κB-FcRn pathway.

Maternal antibiotic treatment during pregnancy attenuates the transport and absorption of maternal antibody IgG through TLR4 and TLR2 receptor.

Maternal antibody IgG, the main antibody in colostrum, plays an important role in neonates protection. Commensal microbiota is closely related to host antibody repertoire. However, there are few reports on how maternal gut microbiota affects maternal antibody IgG transfer. In the present study, we investigated the effects of altering the gut microbiota (treated with antibiotics during pregnancy) on maternal IgG transportation and offspring absorption and explored its underlying mechanisms. Results showed that antibiotic treatment during pregnancy significantly decreased maternal cecal microbial richness (Chao1 and Obesrved species) and diversity (Shannon and Simpson). Plasma metabolome enriched significant changes in the process of bile acid secretion pathway, and the concentration of deoxycholic acid, a secondary metabolite of microorganisms was lowered. Flow cytometry analysis indicated that antibiotic treatment promoted the number of B cells and abated the number of T, DC and M1 cells in intestinal lamina propria of dams. Surprisingly, the serum IgG level in antibiotic treated dams was significantly increased, while IgG contents in colostrum was decreased. Moreover, pregnancy antibiotic treatment in dams was reduced the expression of FcRn, TLR4 and TLR2 in breast of dams and in duodenum and jejunum of neonates. Furthermore, TLR4-/- and TLR2-/- knock-out mice showed a lower FcRn expression in breast of dams and in duodenum and jejunum of neonates. These findings suggest that maternal intestine bacteria may affect the maternal IgG transfer through regulating the breast TLR4 and TLR2 of dams.

Magnolol as a Protective Antioxidant Alleviates Rotenone-Induced Oxidative Stress and Liver Damage through MAPK/mTOR/Nrf2 in Broilers.

This study aimed to investigate the protective effects and molecular mechanism of magnolol supplementation on rotenone-induced oxidative stress in broilers. Two hundred and eighty-eight old male AA broilers were randomly divided into four groups: the CON group: basic diet with sunflower oil injection; the ROT group: basic diet with 24 mg/kg BW rotenone; the MAG + ROT group: basic diet with 300 mg/kg magnolol and rotenone injection; and the MAG group: basic diet with 300 mg/kg magnolol and sunflower oil injection. At 21−27 days of age, the broilers in each group were intraperitoneally injected with rotenone (24 mg/kg BW) or the same volume of sunflower oil. The results showed that magnolol reversed the decrease in ADG post-injection and FBW via rotenone induction. Compared to the ROT group, MAG + ROT group enhanced the average daily gain post injection (p < 0.05). Magnolol supplement could improve the activity and mRNA expression of rotenone-suppressed antioxidant enzymes such as GSH and GSH-PX (p < 0.05). Similarly, the MDA content as an oxidative damage marker was significantly reduced after magnolol addition (p < 0.05). The hepatocyte apoptosis and the mRNA expression of apoptosis-related signaling pathway in the ROT group increased, but magnolol supplementation inhibited rotenone-induced apoptosis through the Nrf2 signaling pathway. Through RNA transcriptome analysis, there were 277 differential genes expressions (DEGs) among the CON group with ROT group, and 748 DEGs were found between the ROT group and the MAG + ROT group. KEGG pathway enrichment found that magnolol relieved rotenone-induced energy metabolism disorder and oxidative damage through signaling pathways such as MAPK and mTOR. In conclusion, magnolol attenuates rotenone-induced hepatic injury and oxidative stress of broilers, presumably by restoring hepatic antioxidant function via the MAPK/mTOR/Nrf2 signaling pathway.

Dietary essential oils improves the growth performance, antioxidant properties and intestinal permeability by inhibiting bacterial proliferation, and altering the gut microbiota of yellow-feather broilers.

This experiment was conducted to investigate the antibacterial effects of essential oils (EO) in vitro and the influence of EO on growth performance, intestinal morphology and oxidation resistance and cecal microflora of yellow-feathered broilers. A total of 720 one-day-old male yellow feather broilers were randomly assigned into 4 treatments with 6 replicate cages of 30 broilers each. The groups were as follows: CON group (basal diet), EO200 group (basal diet + 200 mg/kg EO), EO400 group (basal diet + 400 mg/kg EO), and EO600 group (basal diet + 600 mg/kg EO). The experiment lasted for 48 d. Results showed that the growth and biofilm formation of avian pathogenic E. coli O78 and Salmonella pullorum were limited by adding EO to the diet (P < 0.05). Besides, birds fed with EO had greater (P < 0.05) average daily feed intake (ADFI), average daily gain (ADG), and body weight (BW) during d 1 to 21, 22 to 42, and 1 to 48 and lower (P < 0.05) feed: gain (F:G) than those fed with basal diet during d 22 to 42 and 1 to 48. Moreover, the activity of antioxidant enzyme and the intestinal permeability were improved in the EO400 and EO600 groups rather than the CON group on d 21 (P < 0.05). There were significant differences in cecal microbial composition and enrichment of metabolic pathways of birds among all groups by 16S-based sequencing. In summary, some dose of EO improved bacteriostatic ability, antioxidant ability, and intestinal health of broilers which contributed to the growth performance improvement of yellow-feathered broilers, which can be a promising antibiotic alternative for improving poultry production.

Arbutin improves gut development and serum lipids via Lactobacillus intestinalis.

Arbutin has been widely studied in whitening, anti-inflammatory, and antioxidant. However, the interaction between arbutin and intestinal microbes has been rarely studied. Thus, mice were treated with arbutin concentrations of 0, 0.1, 0.2, 0.4, and 1 mg/ml. We found that arbutin promoted gut development such as villus length, villus areas, and villus length/crypt depth (L/D). Total cholesterol (TC), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were significantly reduced by low concentrations of arbutin. Importantly, we analyzed the microbial composition in the control and 0.4 mg/ml arbutin group and found that the abundance of Lactobacillus intestinalis (L. intestinalis) was highest and enhanced in arbutin. Further, mice were fed with oral antibiotics and antibiotics + 0.4 mg/ml arbutin and then we transplanted fecal microbes from oral 0.4 mg/ml arbutin mice to mice pretreated with antibiotics. Our results showed that arbutin improves gut development, such as villus width, villus length, L/D, and villus areas. In addition, L. intestinalis monocolonization was carried out after a week of oral antibiotics and increased villus length, crypt depth, and villus areas. Finally, in vitro arbutin and L. intestinalis co-culture showed that arbutin promoted the growth and proliferation of L. intestinalis. Taken together, our results suggest that arbutin improves gut development and health of L. intestinalis. Future studies are needed to explore the function and mechanism of L. intestinalis affecting gut development.

Multi-Omics Analysis of the Microbiome and Metabolome Reveals the Relationship Between the Gut Microbiota and Wooden Breast Myopathy in Broilers.

Wooden breast (WB) is a widely prevalent myopathy in broiler chickens. However, the role of the gut microbiota in this myopathy remains largely unknown, in particular the regulatory effect of gut microbiota in the modulation of muscle metabolism. Totally, 300 1-day-old Arbor Acres broilers were raised until 49 days and euthanized, and the breast filets were classified as normal (NORM), mild (MILD), or severe wooden breast (SEV). Birds with WB comprised 27.02% of the individuals. Severe WB filets had a greater L* value, a* value, and dripping loss but a lower pH (P < 0.05). WB filets had abundant myofiber fragmentation, with a lower average myofiber caliber and more fibers with a diameter of <20 µm (P < 0.05). The diversity of the intestinal microflora was decreased in birds with severe WB, with decreases in Chao 1, and observed species indices. At the phylum level, birds with severe WB had a lower Firmicutes/Bacteroidetes ratio (P = 0.098) and a decreased abundance of Verrucomicrobia (P < 0.05). At the species level, gut microbiota were positively correlated with 131 digesta metabolites in pathways of glutamine and glutamate metabolism and arginine biosynthesis but were negatively correlated with 30 metabolites in the pathway of tyrosine metabolism. In plasma, WB induced five differentially expressed metabolites (DEMs), including anserine and choline, which were related to the severity of the WB lesion. The microbial-derived metabolites, including guanidoacetic acid, antiarol, and (2E)-decenoyl-ACP, which entered into plasma were related to meat quality traits and myofiber traits. In summary, WB filets differed in gut microbiota, digesta, and plasma metabolites. Gut microbiota respond to the wooden breast myopathy by driving dynamic changes in digesta metabolites that eventually enter the plasma.

Effects of Different Oligosaccharides on Growth Performance and Intestinal Function in Broilers.

Objective: This study was conducted to investigate the effects of different oligosaccharides on the growth performance and intestinal function in broilers. Methods: A total of 360 1-day-old yellow-feather chickens were randomly divided into 5 groups and fed with a basal diet supplemented with 50 mg/kg chlortetracycline (ANT), 3 g/kg isomalto-oligosaccharide (IMO), 3 g/kg raffinose oligosaccharide (RFO), and 30 mg/kg chitooligosaccharide (COS). The experiment lasted for 56 days, with 1-28 days as the starter phase and 29-56 days as the grower phase. Results: The results showed that dietary supplementation with RFO and COS significantly improved average daily gain (ADG) and average daily feed intake (ADFI) (p < 0.05). Relative to the control group, diets supplemented with oligosaccharides dramatically increased the level of serum IgM (RFO, COS), T-SOD (COS), and GSH-Px (IMO and RFO) and the expression of ZO-1(IMO) and claudin-1 (RFO) (p < 0.05). Adding antibiotics or oligosaccharides to the diet could remarkedly increase the villus height and villus height (VH)/crypt depth (CD) ratio of each group (p < 0.05). Through the ileum α-diversity analysis and comparison of OTU number in each group showed that the microbial richness of the IMO group increased in the starter phase, and that of the RFO and CSO group increased in the grower phase. Additionally, compared with the control group, IMO supplementation increased the level of ileum sIgA (p < 0.05) and the content of valeric acid (p < 0.05) in the cecum. Conclusions: In summary, the addition of oligosaccharides in diet can improve the immune function and antioxidant capacity and improve intestinal health of broilers.

Protective effect and possible mechanism of arctiin on broilers challenged by Salmonella pullorum.

This study was aimed to investigate the effects of dietary arctiin (ARC) supplementation (100, 200, and 400 mg/kg) on the growth performance and immune response of broilers after a Salmonella pullorum (S. pullorum) challenge, and we conducted in vitro antibacterial test to explore the bacteriostatic mechanism of ARC. The in vivo trial was randomly assigned to six groups: noninfected control (NC) group and positive control (PC) group received a basal diet; TET group, received a basal diet supplemented with 100 mg/kg chlortetracycline; ARC100, ARC200, and ARC400 groups received a basal diet containing 100, 200, and 400 mg/kg ARC, respectively. From days 14 to 16, all birds (except the NC group) were infected with 1 mL (1 × 108 CFU per mL) fresh S. pullorum culture by oral gavage per day. In vivo results showed that dietary supplementation of 200 mg/kg ARC significantly increased average daily gain (P < 0.05) and decreased feed-to-gain ratio of broilers vs. the PC group during days 15 to 28 after being challenged with S. pullorum (P < 0.05). The jejunal crypt depth (CD) was decreased by supplementing 100 or 200 mg/kg ARC in diets compared with PC birds at day 19 (P < 0.05). The jejunal villi height (VH) was increased by supplementing 100, 200, or 400 mg/kg ARC in diets compared with PC birds at day 28 (P < 0.05). Besides, dietary supplementation of 200 mg/kg ARC increased the jejunal VH to CD ratio than the PC group both at days 19 and 28 (P < 0.05). Notably, the broilers had lower serum lipopolysaccharide and diamine oxidase levels in the ARC100 and ARC200 groups at day 28 than those in the PC group (P < 0.05). Furthermore, in comparison to PC birds, the birds in ARC groups (100, 200, and 400 mg/kg) had higher serum contents of IgM and IL-10, and the birds in the ARC200 group had higher serum contents of IgA at day 19 (P < 0.05). At day 28, the birds in ARC groups (100, 200, and 400 mg/kg) had lower serum contents of IL-8, and the birds in the ARC200 group had lower serum contents of IFN-γ compared with PC birds (P < 0.05). The in vitro experiment showed that ARC significantly inhibited the biofilm formation and adhesion of S. pullorum (P < 0.05). Metabonomics analysis revealed that ARC can restrain the formation of the biofilm by affecting a variety of metabolic pathways of S. pullorum. Therefore, dietary supplementation of 200 mg/kg ARC might be a potential way to substitute antibiotics to control S. pullorum infection in broilers.

Pullorosis caused by Salmonella pullorum (S. pullorum) is a severe contagious disease and could cause great economic loss to the poultry industry. Antibiotics are usually used to control pullorosis, while prolonged use of antibiotics has led to the emergence of multidrug-resistant bacterial strains. Therefore, it is necessary to find safer and more effective alternatives to substitute antibiotics. In this study, we established a model of S. pullorum infection in broilers and conducted in vitro antibacterial test to explore the preventive effect and mechanisms of dietary arctiin (ARC) supplementation on S. pullorum infection in broilers. The results showed that ARC could not only improve the immune function of infected broilers by regulating the immune system but also directly inhibit the invasion of S. pullorum to broilers by inhibiting the formation and adhesion rate of S. pullorum biofilm. Dietary supplementation of 200 mg/kg ARC might be a potential way to substitute antibiotics to control S. pullorum infection in broilers.

The effects of magnolol supplementation on growth performance, meat quality, oxidative capacity, and intestinal microbiota in broilers.

The aim of the present study was to evaluate the effect of magnolol (MAG) on growth performance, meat quality, oxidative capacity, and intestinal microbiota in the yellow-feather broilers. A total of 360 one-day-old male yellow-feather broiler chicks were allocated into 5 groups of 6 replicates and 12 chickens per replicate, were fed a basal diet supplemented with 0 (Control group, CON), 100, 200, 300, or 400 mg/kg MAG for 51 d. The results showed that dietary supplementation with 200 and 300 mg/kg MAG increased the average daily gain (ADG) but decreased feed to gain ratio (F/G) during the overall periods (P < 0.05). Dietary MAG increased significantly the redness value (a∗) of the breast muscle (P < 0.05), and decreased the water loss rate and shear force of the breast meat (P < 0.05). MAG supplement reduced the malondialdehyde (MDA) levels, and increased the glutathione (GSH), superoxide dismutase (SOD), and total antioxidant capacity (T-AOC) levels in breast muscle and jejunum. PCR analysis showed that MAG increased the levels of NF-E2-related factor 2 (Nrf2), NAD(P)H/quinone oxidoreductase 1 (NQO1), heme oxygenase-1 (HO-1), glutathione-S transferase (GST), glutamate-cysteine ligase catalytic subunit (GCLC), glutamate-cysteine ligase modifier subunit (GCLM), and SOD expressions (P < 0.05). Analysis of differential enrichment of gut microbiota found that Faecalibacterium in the cecum of MAG supplemented broilers increased, and Coprobacillus has decreased (P < 0.05). In conclusion, MAG improved growth performance, meat quality of the broilers and antioxidant capacity, and modulated gut microbiota homeostasis.

Dietary Beta-Hydroxy-Beta-Methyl Butyrate Supplementation Inhibits Hepatic Fat Deposition via Regulating Gut Microbiota in Broiler Chickens.

The present study is aimed to explore the effects of different dietary beta-hydroxy-beta-methyl butyrate (HMB) levels (0, 0.05%, 0.10%, or 0.15%) on liver lipid metabolism on Wenshi broiler chickens. Results showed that HMB reduced the liver weight as well as liver concentrations of triacylglycerol (TG) and total cholesterol (TC) (quadratically, p < 0.05), and the lowest values were observed in the 0.10% HMB group. Meanwhile, HMB supplementation significantly altered the expression levels of key genes related to lipid metabolism in the liver of broiler chickens (p < 0.05). Furthermore, 16S rRNA gene sequencing revealed that HMB supplementation could greatly change the richness, diversity, and composition of the broiler gut microbiota, and the Bacteroidetes relative abundance at the phylum level and the Alistipes relative abundance at the genus level were affected (p < 0.05). Correlation analysis further suggested a strong association between Bacteroidetes relative abundance and lipid metabolism-related parameters (p < 0.05). Together, these data suggest that 0.10% HMB supplementation could inhibit hepatic fat deposition via regulating gut microbiota in broilers.

The preventive effect and mechanisms of adsorbent supplementation in low concentration aflatoxin B1 contaminated diet on subclinical symptom and histological lesions of broilers.

This study aimed to investigate the subclinical symptom and histological lesions of 21-day-old and 42-day-old broilers exposure to low concentration aflatoxin B1 (AFB1), and the preventive effect with adsorbent (Toxo-MX) supplementation. A total of 576 one-day-old Arbor Acres broilers were randomly allotted into 6 treatments 8 replicates and 12 birds per cage, fed with 0 ppb, 60 ppb and 120 ppb AFB1 contamination diet with or without Toxo-MX supplementation. Results showed both 60 ppb and 120 ppb AFB1 contamination significantly reduced growth performance in 21-day-old broilers (P < 0.05), but not in 42-day-old broilers (P > 0.05), however, AFB1 contamination in diet caused a higher feed to gain ratio (P < 0.05). Broilers of 21-day-old exposure to 60 ppb and 120 ppb AFB1 increased mRNA expression of hepatic inflammatory cytokines, and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activity (P < 0.05), 42-day-old broilers showed a same change in 120 ppb but not in 60 ppb of AFB1 contamination (P < 0.05). mRNA expressions of clauding-1, Zonula occludens-1 (ZO-1), and occludin decreased, but Bax, Bcl-2, and caspase-3 increased in 21-day-old broilers exposure to 60 ppb and 120 ppb AFB1 (P < 0.05), broilers of 42-day-old resisted on intestinal aflatoxicosis impairment against 60 ppb AFB1 contamination (P < 0.05), but not in 120 ppb (P < 0.05). Toxo-MX supplementation significantly reversed the detrimental effects on growth performance in both age broilers and reduced the accelerated feed to gain ratio caused by AFB1 (P < 0.05). Intestinal mRNA expression of tight junction and apoptotic genes in both age broilers were recovered by Toxo-MX supplementation (P < 0.05). However, Toxo-MX did not restore the accelerated expression of hepatic inflammation cytokines and SOD, GSH-Px in 120ppb AFB1 group (P < 0.05). The data demonstrated that diet supplementation with Toxo-MX reversed the detrimental effect on growth performance and intestine in broilers exposure to 60 ppb and 120 ppb AFB1. However, did not completely recovered hepatic inflammation induced by AFB1.

Effects of Dietary Ginsenoside Rg1 Supplementation on Growth Performance, Gut Health, and Serum Immunity in Broiler Chickens.

The restriction and banning of antibiotics in farm animal feed has led to a search for promising substitutes for antibiotics to promote growth and maintain health for livestock and poultry. Ginsenoside Rg1, which is one of the most effective bioactive components in ginseng, has been reported to have great potential to improve the anti-inflammatory and anti-oxidative status of animals. In this study, 360 Chinese indigenous broiler chickens with close initial body weight were divided into 5 groups. Each group contained 6 replicates and each replicate had 12 birds. The experimental groups were: the control group, fed with the basal diet; the antibiotic group, fed basal diet + 300 mg/kg 15% chlortetracycline; and three Rg1 supplementation groups, fed with basal diet + 100, 200, and 300 mg/kg ginsenoside Rg1, respectively. The growth performance, immune function, and intestinal health of birds were examined at early (day 1-28) and late (day 29-51) stages. Our results showed that dietary supplementation of 300 mg/kg ginsenoside Rg1 significantly improved the growth performance for broilers, particularly at the late stage, including an increase in final body weight and decrease of feed conversion ratio (P < 0.05). Additionally, the integrity of intestinal morphology (Villus height, Crypt depth, and Villus height/Crypt depth) and tight junction (ZO-1 and Occludin), and the secretion of sIgA in the intestine were enhanced by the supplementation of Rg1 in chicken diet (P < 0.05). The immune organ index showed that the weight of the thymus, spleen, and bursa was significantly increased at the early stage in ginsenoside Rg1 supplementation groups (P < 0.05). Our findings might demonstrate that ginsenoside Rg1 could serve as a promising antibiotic alternative to improve the growth performance and gut health for broiler chickens mainly through its amelioration of inflammatory and oxidative activities.

Protective Effects of L-Theanine on IPEC-J2 Cells Growth Inhibition Induced by Dextran Sulfate Sodium via p53 Signaling Pathway.

L-theanine is a nonprotein amino acid found in tea leaves and has been widely used as a safe food additive in beverages or foods because of its varied bioactivities. The aim of this study was to reveal the in vitro gastrointestinal protective effects of L-theanine in DSS-induced intestinal porcine enterocyte (IPEC-J2) cell models using molecular and metabolic methods. Results showed that 2.5% dextran sulfate sodium (DSS) treatment inhibited the cell proliferation of IPEC-J2 and blocked the normal operation of the cell cycle, while L-theanine pretreatment significantly preserved these trends to exert protective effects. L-theanine pre-treatment also up-regulated the EGF, CDC2, FGF2, Rb genes and down-regulated p53, p21 proliferation-related mRNA expression in DSS-treated cells, in accompany with p53 signaling pathway inhibition. Meanwhile, metabolomics analysis revealed that L-theanine and DSS treated IPEC-J2 cells have different metabolomic profiles, with significant changes in the key metabolites involved in pyrimidine metabolism and amino acid metabolism, which play an important role in nucleotide metabolism. In summary, L-theanine has a beneficial protection in DSS-induced IPEC-J2 cells via promoting proliferation and regulating metabolism disorders.

Effects of Feeding Fermented Mulberry Leaf Powder on Growth Performance, Slaughter Performance, and Meat Quality in Chicken Broilers.

This study was conducted to investigate the effects of feeding fermented mulberry leaf powder (FMLP) on growth performance, slaughter performance, and meat quality of broilers. A total of 360 1-day-old chickens were randomly divided into 5 groups. The control group was fed basal diet (CON), 3% FMLP, 6% FMLP, 9% FMLP, and 3% unfermented mulberry leaf powder. The (MLP) group was fed basal diet supplemented with 3%, 6%, 9% fermented mulberry leaf powder, and 3% MLP, respectively. The experiment lasted for 56 days, with 1-28 days as the starter phase and 29-56 days as the grower phase. The results on the growth performance showed that diets supplemented with 3% FMLP significantly increased the ratio of villus height to crypt depth in the duodenum, jejunum, and ileum of broilers, enhanced the activity of intestinal amylase and digestibility of dry matter and crude protein, improved the average daily gain (ADG), and decreased the feed to gain ratio (F/G) (p < 0.05). Compared with the control group diet, the 3% FMLP group diet significantly increased the breast muscle yield (p < 0.05), reduced the abdominal fat ratio (0.1 < p < 0.05), and improved the slaughter performance of broilers. The 3% MLP group diet increased the shear force of breast muscle (p < 0.05) and thigh muscle of broilers compared to the control group, and adding FMLP could reverse the above results. Additionally, relative to the control group, FMLP supplementation improved the contents of inosine monophosphate (IMP), total amino acids (TAA), essential amino acids (EAA), and delicious amino acids (DAA) in breast and thigh muscle, and improved polyunsaturated fatty acids (PUFA) and essential fatty acids (EFA) in breast muscle; the 6% and 9% FMLP groups showed preferably such effects (p < 0.05). In conclusion, dietary supplementation of FMLP can improve the digestion and absorption of nutrients, and then improve the growth performance of broilers; it also has a positive effect on improving slaughter performance and meat quality.

Protective Effect of l-Theanine against DSS-Induced Colitis by Regulating the Lipid Metabolism and Reducing Inflammation via the NF-κB Signaling Pathway.

The present study revealed the phylactic effects of l-theanine on a DSS-induced colitis mice model. The results showed that 3% DSS treatment significantly induced intestinal damage as reflected by DAI, histopathological feature, and colon length, while l-theanine pretreatment markedly prevented these trends to exert protective effects. Meanwhile, l-theanine pretreatment decreased the levels of TNF-α, IL-1ß, IL-6, iNOS, and COX2 on DSS-induced colitis. Notably, DSS inhibited the proliferation and promoted the apoptosis of intestinal epithelial cells, thereby damaging the integrity of the intestinal epithelial barrier, whereas l-theanine also played a protective role by attenuating these deteriorated effects. It was also observed that l-theanine treatment downregulated the levels of p-p65, p65, p-p53, p53, and p-AKT protein expression in acute DSS-induced colitis, which showed the protective function of l-theanine, mainly via the NF-κB signaling pathway. Furthermore, the results of lipid analysis and transcriptome analysis show that l-theanine reversed transcriptional profiles and lipid profiles of colitis models, mainly via the inflammatory reactivity-related pathway. Interestingly, the correlation analysis between transcriptional profiles and lipid profiles showed that inflammatory response-related genes were almost significantly correlated with differential lipid metabolites. In summary, l-theanine plays a protective role in DSS-induced colitis via downregulating the NF-κB signaling pathway and regulating lipid metabolism disorders.

Eugenol Alleviates Dextran Sulfate Sodium-Induced Colitis Independent of Intestinal Microbiota in Mice.

The present study investigated the effect of eugenol (EUG) on dextran sulfate sodium (DSS)-induced colitis and explored the underlying mechanisms. C57BL/6 mice were intragastrically administered normal saline or EUG (20 mg/kg body weight) for 17 days, and colitis was induced by using 3% DSS from day 7. The results showed that EUG increased the body weight and reduced the disease activity index score and colon pathological scores in DSS-treated mice (P < 0.05). Further, EUG preserved the proinflammatory cytokines (interleukin (IL)-6, -12, -21, and -23), lowered (P < 0.05) colonic malondialdehyde (MDA), uncoupling protein 2 (UCP2) expression and p65 phosphorylation, and activated (P < 0.05) colonic kelch-like ECH-associated protein 1 and nuclear factor (erythroid-derived 2)-like 2 expressions but did not affect the intestinal microbiota in DSS-treated mice. Furthermore, EUG ameliorated colitis in antibiotic-treated mice, while fecal microbiota transplantation from EUG preadministered mice failed to ameliorate colitis. In conclusion, EUG could alleviate colitis by attenuating colonic inflammation and oxidative stress independent of intestinal microbiota.