Magnolol-driven microbiota modulation elicits changes in tryptophan metabolism resulting in reduced skatole formation in pigs.

Skatole of gut origin has garnered significant attention as a malodorous pollutant due to its escalating emissions, recalcitrance to biodegradation and harm to animal and human health. Magnolol is a health-promoting polyphenol with potential to considerably mitigate the skatole production in the intestines. To investigate the impact of magnolol and its underlying mechanism on the skatole formation, in vivo and in vitro experiments were conducted in pigs. Our results revealed that skatole concentrations in the cecum, colon, and faeces decreased by 58.24% (P = 0.088), 44.98% (P < 0.05) and 43.52% (P < 0.05), respectively, following magnolol supplementation. Magnolol supplementation significantly decreased the abundance of Lachnospira, Faecalibacterium, Paramuribaculum, Faecalimonas, Desulfovibrio, Bariatricus, and Mogibacterium within the colon (P < 0.05). Moreover, a strong positive correlation (P < 0.05) between skatole concentration and Desulfovibrio abundance was observed. Subsequent in silico studies showed that magnolol could dock well with indolepyruvate decarboxylase (IPDC) within Desulfovibrio. Further in vitro investigation unveiled that magnolol addition led to less indole-3-pyruvate diverted towards the oxidative skatole pathway by the potential docking of magnolol towards IPDC, thereby diminishing the conversion of substrate into skatole. Our findings offer novel targets and strategies for mitigating skatole emission from the source.

Magnolol Supplementation Alters Serum Parameters, Immune Homeostasis, Amino Acid Profiles, and Gene Expression of Amino Acid Transporters in Growing Pigs.

This study investigated whether dietary supplementation with magnolol affects growth performance, anti-inflammatory abilities, serum and muscle amino acid profiles, and metabolisms in growing pigs. A total of 42 seventy-days-old growing barrows (Duroc × Landrace × Yorkshire) were randomly allocated into two dietary groups: Con, control group (basal diet); and Mag, magnolol group (basal diet supplemented with 400 mg/kg of magnolol). The results revealed that dietary supplementation with magnolol had no effect (p > 0.05) on growth performance. However, magnolol supplementation remarkably increased (p < 0.05) the serum content of albumin, total protein, immunoglobulin G, immunoglobulin M, and interleukin-22. In addition, dietary magnolol supplementation altered the amino acid (AA) profiles in serum and dorsal muscle and particularly increased (p < 0.05) the serum content of arginine and muscle glutamate. Simultaneously, the mRNA expression of genes associated with AA transport in jejunum (SLC38A2, SLC1A5, and SLC7A1) and ileum (SLC1A5 and SLC7A1) was higher (p < 0.05) in the Mag group than in the Con group. Additionally, the serum metabolomics analysis showed that the addition of magnolol significantly enhanced (p < 0.05) arginine biosynthesis, as well as D-glutamine and D-glutamate metabolism. Overall, these results suggested that dietary supplementation with magnolol has the potential to improve the accumulation of AAs, protein synthesis, immunity, and body health in growing pigs by increasing intestinal absorption and the transport of AAs.

Density Functional Study to Investigate the Ability of (ZnS)n (n = 1-12) Clusters Removing Hg0, HgCl, and HgCl2 via Electron Localization Function and Non-Covalent Interactions Analyses.

In this study, the density functional theory is used to study the ability of (ZnS)n clusters to remove Hg0, HgCl, and HgCl2 and reveals that they can be absorbed on (ZnS)n clusters. According to electron localization function (ELF) and non-covalent interactions (NCI) analyses, the adsorption of Hg0 on (ZnS)n is physical adsorption and the adsorption ability of (ZnS)n for removing Hg0 is weak. When (ZnS)n adsorbs HgCl and HgCl2, two new Hg-S and Zn-Cl bonds form in the resultant clusters. An ELF analysis identifies the formation of Hg-S and Zn-Cl bonds in (ZnS)nHgCl and (ZnS)nHgCl2. A partial density of states and charge analysis confirm that as Hg0, HgCl, and HgCl2 approach (ZnS)n clusters, atomic orbitals in Hg and Zn, Hg and S, as well as Zn and Cl overlap and hybridize. Adsorption energies of HgCl and HgCl2 on (ZnS)n clusters are obviously bigger than those of Hg0, indicating that HgCl and HgCl2 adsorption on (ZnS)n clusters is much stronger than that of Hg0. By combining ELF analysis, NCI analysis, and adsorption energies, the adsorption of HgCl, and HgCl2 on (ZnS)n clusters can be classified as chemical adsorption. The adsorption ability of (ZnS)n clusters for removing HgCl and HgCl2 is higher than that of Hg0.

Mulberry leaf extract improves intestinal barrier function and displays beneficial effects on colonic microbiota and microbial metabolism in weaned piglets.

BACKGROUND: Mulberry leaf extract (MLE) extracted from mulberry leaves is rich in a variety of bioactive ingredients and can be used as feed additives of weaned piglets. The present study was conducted to evaluate the effects of dietary MLE supplementation on intestinal barrier function, colon microbial numbers and microbial metabolites of weaned piglets. RESULTS: MLE supplementation increased the villus height and the villus height/crypt depth ratio in jejunum and ileum (P < 0.05), increased the mRNA expression of ZO-1, Claudin-1 and MUC-2 in the ileal mucosa (P < 0.05), and decreased the serum level of lipopolysaccharide (P < 0.01). Meanwhile, MLE reduced the mRNA expression of tumor necrosis factor-α and interleukin-1ß (P < 0.05) and increased secretory immunoglobulin A level in the ileal mucosa (P < 0.05). In addition, MLE increased the numbers of beneficial bacteria Bifidobacterium and Lactobacillus (P < 0.05) and decreased the number of potential pathogenic bacteria Escherichia coli (P < 0.05) in the colon. Correspondingly, MLE supplementation reduced the pH value of colonic digesta (P < 0.05) and altered the microbial fermentation pattern of the colon by increasing the concentrations of microbial metabolites derived from carbohydrates fermentation such as lactate, acetate, butyrate and total short-chain fatty acids (P < 0.05), and decreasing the concentrations of microbial metabolites derived from amino acid fermentation such as p-cresol, skatole, spermine, histamine and tryptamine (P < 0.05). CONCLUSION: MLE supplementation improved intestinal barrier function and displayed beneficial effects on colon microbes and microbial metabolism in weaned piglets. © 2022 Society of Chemical Industry.

Guanidine acetic acid supplementation altered plasma and tissue free amino acid profiles in finishing pigs.

BACKGROUND: As a nutritive feed additive, guanidine acetic acid (GAA) participates in the metabolism of energy and proteins. This study aimed to investigate the effects of GAA on growth performance, organ index, plasma and tissue free amino acid profiles, and related metabolites in finishing pigs. A total of 72 crossbred pigs (body weight 86.59 ± 1.16 kg) were randomly assigned to 1 of 4 dietary treatments (GAA0, GAA500, GAA1000, and GAA1500). They were fed the basal diets supplemented with 0, 500, 1000, or 1500 mg/kg GAA for 42 days, respectively. The growth performance and organ weight were evaluated, and the contents of crude protein, free amino acids, and metabolites in plasma and tissues were determined. Spearman correlation between plasma and tissue free amino acids and related metabolites was also analyzed. RESULTS: Growth performance in pigs was not altered by GAA (P > 0.05). The absolute and relative weight of kidneys increased (quadratic, P < 0.05). As dietary GAA concentration was increased, the contents of plasma glycine, serine, leucine, ornithine, and ratio of ornithine/arginine decreased (linear or quadratic, P < 0.05), but the contents of plasma isoleucine and taurine and the ratios of alanine/branched-chain amino acids and proline/ornithine increased quadratically (P < 0.05). The hepatic γ-amino-n-butyric acid content increased linearly and quadratically (P < 0.001), while the carnosine content decreased (quadratic, P = 0.004). The contents of renal arginine, proline, cystine, glutamate, and total amino acids (TAA) decreased quadratically (P < 0.05), but the contents of glycine (quadratic, P = 0.015) and γ-amino-n-butyric acid (linear, P = 0.008) increased. The pancreatic tryptophan content (quadratic, P = 0.024) increased, while the contents of pancreatic proline (linear, P = 0.005) and hydroxyproline (quadratic, P = 0.032) decreased in response to GAA supplementation. The contents of cardiac essential amino acids (EAA), nonessential amino acids (NEAA), and TAA in GAA1000 were higher than those in GAA1500 (P < 0.05). Supplementing with GAA linearly increased the contents of methionine, threonine, valine, isoleucine, leucine, phenylalanine, tryptophan, lysine, histidine, arginine, serine, alanine, glutamine, asparagine, tyrosine, proline, taurine, cystathionine, α-aminoadipic acid, ß-aminoisobutyric acid, EAA, NEAA, and TAA in the spleen (P < 0.05). A strong Spearman correlation existed between plasma and tissue free amino acids and related metabolites. CONCLUSION: GAA supplementation did not altered pig growth performance, but it altered plasma and tissue free amino acid profiles and the contents of related metabolites in pigs in a tissue-dependent manner.

Functional Characterization and Whole-Genome Analysis of an Aflatoxin-Degrading Rhodococcus pyridinivorans Strain.

Aflatoxin B1 (AFB1) is one of the most toxic, naturally occurring carcinogen compounds and is produced by specific strains of fungi. Crop contamination with AFB1 can cause huge economic losses and serious health problems. Many studies have examined the microbiological degradation of AFB1, especially the use of efficient AFB1-degrading microorganisms, to control AFB1 contamination. Here, we reported the identification of a new Rhodococcus pyridinivorans strain (4-4) that can efficiently degrade AFB1 (degradation rate 84.9%). The extracellular component of this strain showed the strongest capacity to degrade AFB1 (degradation rate 83.7%). The effects of proteinase K, SDS, temperature, pH, incubation time, and AFB1 concentration on the AFB1 degradation ability of the extracellular component were investigated. We sequenced the complete genome of this strain, encoding 5246 protein-coding genes and 169 RNA genes on a circular chromosome and two plasmids. Comparative genomic analysis revealed high homology with other Rhodococcus strains with high AFB1-degradation ability. Further proteomic analyses of this strain identified a total of 723 proteins in the extracellular component, including multiple potential AFB1-degrading enzymes, along with enzymes that are reported to response to AFB1 treatment. Overall, the results demonstrate that R. pyridinivorans 4-4 would be an excellent candidate for the biodegradation and detoxification of AFB1 contamination.

Exogenous Glutathione Protects IPEC-J2 Cells against Oxidative Stress through a Mitochondrial Mechanism.

The accumulation of reactive oxygen species (ROS) triggers oxidative stress in cells by oxidizing and modifying various cellular components, preventing them from performing their inherent functions, ultimately leading to apoptosis and autophagy. Glutathione (GSH) is a ubiquitous intracellular peptide with multiple functions. In this study, a hydrogen peroxide (H2O2)-induced oxidative damage model in IPEC-J2 cells was used to investigate the cellular protection mechanism of exogenous GSH against oxidative stress. The results showed that GSH supplement improved the cell viability reduced by H2O2-induced oxidative damage model in IPEC-J2 cells in a dose-dependent manner. Moreover, supplement with GSH also attenuated the H2O2-induced MMP loss, and effectively decreased the H2O2-induced mitochondrial dysfunction by increasing the content of mtDNA and upregulating the expression TFAM. Exogenous GSH treatment significantly decreased the ROS and MDA levels, improved SOD activity in H2O2-treated cells and reduced H2O2-induced early apoptosis in IPEC-J2 cells. This study showed that exogenous GSH can protect IPEC-J2 cells against apoptosis induced by oxidative stress through mitochondrial mechanisms.

Effects of dietary citrus extract on growth performance, carcass characteristics and meat quality of pigs.

This study investigated the effects of citrus extract on growth, carcass and meat quality of Duroc × Landrace × Large White pigs. One hundred and eight pigs (54 barrows, 54 females) were assigned to one of three dietary treatments for 138 days. The dietary treatments were (1) basic diet; (2) basic diet supplemented with 75 mg/kg chlortetracycline; and (3) basic diet supplemented with citrus extract (0.25 ml/kg during 56-112 days of age and 0.20 ml/kg during 113-194 days of age). No significant differences among treatments were found for growth performance, carcass characteristics, meat quality and free amino acids (p > 0.05). Feeding citrus extract tended to increase intramuscular fat (p = 0.052). Citrus extract and chlortetracycline increased C15:0 concentration (p = 0.016) and superoxide dismutase activity (p = 0.004). The pigs that received chlortetracycline exhibited the lowest (p = 0.033) muscle malondialdehyde concentration. Overall, citrus extract ameliorated some meat quality indicators without adverse effects on pig growth or carcass performance.

Solid-state fermentation by Aspergillus niger and Trichoderma koningii improves the quality of tea dregs for use as feed additives.

This study evaluated the ability of Aspergillus niger and Trichoderma koningii to improve the quality of tea dregs (TDs) through solid-state fermentation as well as the value of the fermented tea dregs (FTDs) produced for use as bio-feed additives. After fermentation, FTDs differed in color and structure. Fermentation with A. niger and T. koningii increased the contents of crude protein, crude fiber, neutral detergent fiber, and acid detergent fiber of TDs. Compared to the unfermented group, the contents of reducing sugar, total flavonoids, total polyphenols, and theasaponins were increased in A. niger FTDs, while in T. koningii FTDs caffeine was completely degraded, the theasaponins were lower, and the contents of reducing sugar and caffeine higher. Regarding free amino acids, A. niger FTDs had the highest content of total amino acids, total essential amino acids, total non-essential amino acids, total aromatic amino acids, total branched-chain amino acids, and total non-protein amino acids, and all types of essential amino acids, followed by T. koningii FTDs and the control TDs. Fungal fermentation had similar effects on the content of various hydrolytic amino acids as those on above free amino acids, and increased the content of bitter and umami components. The composition of essential amino acids of TDs or FTDs was similar to that of the standard model, except for sulfur-containing amino acids and isoleucine. Solid-state fermentation with A. niger and T. koningii effectively improved the nutritional value of TDs, increased the contents of functional substances, and improved the flavor of TDs. This study demonstrated a feasible approach to utilize TDs that not only increases animal feed resources, but also reduces the production of resource waste and pollution.

Chlorogenic acid: A comprehensive review of the dietary sources, processing effects, bioavailability, beneficial properties, mechanisms of action, and future directions.

Chlorogenic acids (CGAs), a group of hydroxycinnamates, are generally abundant in everyday foods and beverages, most prominently in certain coffee drinks. Among them, the chlorogenic acid (CGA), also termed as 5-O-caffeoylquinic acid (5-CQA), is one of the most abundant, highly functional polyphenolic compounds in the human diet. The evidence of its health benefits obtained from clinical studies, as well as basic research, indicates an inverse correlation between 5-CQA consumption and a lower risk of metabolic syndromes and chronic diseases. This review focuses on the beneficial properties for health and mechanisms of action of 5-CQA, starting with its history, isomers, dietary sources, processing effects, preparation methods, pharmacological safety evaluation, and bioavailability. It also provides the possible molecular mechanistic bases to explain the health beneficial effects of 5-CQA including neuroprotective, cardiovascular protective, gastrointestinal protective, renoprotective, hepatoprotective, glucose and lipid metabolism regulatory, and anticarcinogenic effects. The information summarized here could aid in the basic and clinical research on 5-CQA as a natural dietary additive, potential drug candidate, as well as a natural health promoter.

Effect of amino acids and their derivatives on meat quality of finishing pigs.

Amino acids provide key nutritional value, and significantly contribute to taste and flavor of meat. Here, we review the role of free amino acids in the muscle fibers in meat quality and sensory signals. We furthermore discuss how dietary supplementation of free amino acids and their derivatives (e.g. tryptophan, threonine, arginine, lysine, leucine, glutamate, threonine, sarcosine, betaines, and cysteamine) can influence these attributes. The available data shows that the quality of the meat is subject to the amino acids that are provided in the animal feed.

Equol Inhibits LPS-Induced Oxidative Stress and Enhances the Immune Response in Chicken HD11 Macrophages.

BACKGROUND/AIMS: There has been increasing recent attention on the antioxidative capacity of equol. This study tested the effect of equol on oxidative stress induced by lipopolysaccharide (LPS) and regulation of immunity in chicken macrophages. METHODS: Chicken HD11 macrophages were challenged with LPS (100 ng/mL) alone or with LPS (100 ng/mL) and (±)equol (10, 20, 40, 80, 160 µmol/L) together for 24h. Evaluated responses included the contents of malondialdehyde (MDA) and reduced glutathione (GSH), activities of total superoxide dismutase (T-SOD) and inducible nitric oxide synthase (iNOS), transcript abundance of superoxide dismutase 2 (SOD2), catalase (CAT), glutathione transferase (GST), Toll-like receptor 4 (TLR4), tumor necrosis factor alpha (TNFα) and interleukin-1 beta (IL-1ß), and contents of the cytokines TNFα, IL-1ß, interleukin-2 (IL-2) and interferon beta (IFNß). RESULTS: Exposure to LPS induced oxidative stress as contents of MDA increased and GSH decreased in LPS-treated cells (P < 0.05) compared to those in control cells. Compared to LPS alone, co-treatment with equol (20 µmol/L, 40 µmol/L or 80 µmol/L) reduced contents of MDA and increased those of GSH (both P < 0.05). Activity of T-SOD increased (P < 0.05) in cells treated with the higher contentration of equol (80 µmol/L or 160 µmol/L), however, all concentrations (20 µmol/L to 160 µmol/L) increased activity of iNOS (P < 0.05). The highest concentration of equol (160 µmol/L) increased SOD2 and GST transcripts (P < 0.05). Equol treatment increased transcripts of TLR4, TNFα and IL-1ß (P < 0.05). And there were similar changes in contents of IL-1ß, IL-2, IFNß and TNFα in the cells (P < 0.05). CONCLUSIONS: It concluded that equol can protect chicken HD11 macrophages from oxidative stress induced by LPS through reducing lipid peroxidation products and enhancing contents of antioxidants, and activities of relevant antioxidase enzymes; effects were also seen in gene expression related to the immune response and increased contents of cytokines. The optimal concentration of equol on antioxidation and immune enhancement in chicken macrophages was 40 µmol/L.

Transferrin-conjugated nanodiamond as an intracellular transporter of chemotherapeutic drug and targeting therapy for cancer cells.

AIM: PEGylated fluorescent nanodiamond (FND) conjugated with Tf (FND-PEG-Tf) was investigated for targeted drug delivery. MATERIALS & METHODS: Human hepatoma (HepG2) and normal (L-02) cell lines were used to investigate the difference in cellular uptake of FND-PEG-Tf and its loading drug system. Nanoparticle uptake was evaluated by flow cytometry and laser scanning confocal microscopy. RESULTS: FND-PEG-Tf showed highly specific TfR-mediated uptake by HepG2 cells, relative to negative controls (L-02 cell), which was a strong correlation among TfR density on the cell surface. The mechanism of TfR-mediated uptake was attested by free Tf with Fe³âº as a competitive agent. The difference in cell viability between L-02 and HepG2 cells treated with doxorubicin hydrochloride (DOX) nanoparticles (FND-PEG-Tf-DOX) can be explained by FND-PEG-Tf, which can target drug delivery to cancer cells. CONCLUSION: FND-PEG-Tf can potentially be utilized in targeted cancer cell imaging and effective drug delivery for cancer therapy.

In vivo enhancement of anticancer therapy using bare or chemotherapeutic drug-bearing nanodiamond particles.

BACKGROUND: This study investigated the use of nanodiamond particles (NDs) as a promising material for drug delivery in vivo and in vitro. METHODS: HepG2 cells (a human hepatic carcinoma cell line) were used to determine the characteristics of a nanodiamond-doxorubicin complex (ND-DOX) when taken up by cells in vitro using laser scanning confocal microscopy and dialysis experiments. We also compared the survival rate and histopathology of tumor-bearing mice after treatment with NDs or ND-DOX in vivo. RESULTS: In vitro investigation showed that ND-DOX has slow and sustained drug release characteristics compared with free doxorubicin. In vivo, the survival rate of tumor-bearing mice treated with ND-DOX was four times greater than that of mice treated with free doxorubicin. Interestingly, the survival rate in mice treated with NDs alone was close to that of mice treated with free doxorubicin. This indicates that treatment with ND-DOX can prolong the lifespan of tumor-bearing mice significantly compared with conventional doxorubicin and that NDs can have this effect as well. Histopathological analysis showed that neither the NDs nor ND-DOX were toxic to the kidney, liver, or spleen in contrast with the well-known toxic effects of free doxorubicin on the kidney and liver. Further, both the bare NDs and ND-DOX could suppress tumor growth effectively. CONCLUSION: NDs can potentially prolong survival, and ND-DOX may act as a nanodrug with promising chemotherapeutic efficacy and safety.