Antioxidant Enzyme System Modulation by Dietary Palm Oils, Palm Kernel Oil and Soybean Oil in Laying Hens.

Palm-based oils (palm oil and kernel oil) and soybean oil have unique fatty acid and antioxidant profiles based on the compounds present in them. Hence, this study elucidated the antioxidant properties of crude palm oil (CPO), red palm oil (RPO), refined palm oil (RBD), palm kernel oil (PKO) and soybean oil (SBO) and the influence of dietary oils on blood lipid profiles, tissue fatty acid deposition and the expression of hepatic lipid and lipoprotein metabolism genes in laying hens. The oils were analyzed for color, beta-carotene, free fatty acid and acid value, phenolic content and lipid peroxidation. In an in vivo trial, 150 laying hens were allotted into five groups and supplemented with either CPO, RPO, RBD, PKO or SBO for 16 weeks. High antioxidant compounds present in palm oils help reduce the oxidation of oils. Dietary supplementation with palm oils, particularly CPO and RPO, contributed to the lower liver, serum and jejunal mucosal antioxidant enzyme activities. The antioxidant enzyme genes in the jejunal mucosa were downregulated in palm oils and PKO, but there was no difference between oils in antioxidant enzyme genes in the liver. In conclusion, dietary supplementation with oils with high antioxidant content contributed to protection against oxidation and was associated with a lower requirement for producing antioxidant enzymes.

Brown and Green Seaweed Antioxidant Properties and Effects on Blood Plasma Antioxidant Enzyme Activities, Hepatic Antioxidant Genes Expression, Blood Plasma Lipid Profile, and Meat Quality in Broiler Chickens.

The study was designed to analyze the effects of brown seaweed (BS) and green seaweed (GS) on blood plasma antioxidant enzyme activities, hepatic antioxidant genes expression, blood plasma lipid profile, breast meat quality, and chemical composition in broiler chickens. The dietary treatment groups contained basal diet [negative control (NC)], basal diet + vitamin E (100 mg/kg feed) [positive control (PC)], basal diet + 0.25, 0.50, 0.75, 1, and 1.25% BS and GS supplements separately. The findings showed that both BS and GS exhibited remarkable antioxidant activity. In contrast, the maximum antioxidant activity was recorded by BS (55.19%), which was significantly higher than the GS (25.74%). Results showed that various levels of BS and GS had no significant effects on broiler blood plasma catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) enzyme activities. The hepatic superoxide dismutase 1 (SOD1) gene mRNA expression was significantly higher for birds fed 0.50% and 0.75% BS. Regarding the plasma lipid profile, the total cholesterol (TC) and high-density lipoprotein (HDL) levels were higher (p < 0.05) for birds fed 0.75 and 1% BS compared to the negative and positive control groups. The findings showed that different levels of BS and GS had significantly higher breast meat crude protein (CP) content.

A refined medium to enhance the antimicrobial activity of postbiotic produced by Lactiplantibacillus plantarum RS5.

Postbiotic RS5, produced by Lactiplantibacillus plantarum RS5, has been identified as a promising alternative feed supplement for various livestock. This study aimed to lower the production cost by enhancing the antimicrobial activity of the postbiotic RS5 by improving the culture density of L. plantarum RS5 and reducing the cost of growth medium. A combination of conventional and statistical-based approaches (Fractional Factorial Design and Central Composite Design of Response Surface Methodology) was employed to develop a refined medium for the enhancement of the antimicrobial activity of postbiotic RS5. A refined medium containing 20 g/L of glucose, 27.84 g/L of yeast extract, 5.75 g/L of sodium acetate, 1.12 g/L of Tween 80 and 0.05 g/L of manganese sulphate enhanced the antimicrobial activity of postbiotic RS5 by 108%. The cost of the production medium was reduced by 85% as compared to the commercially available de Man, Rogosa and Sharpe medium that is typically used for Lactobacillus cultivation. Hence, the refined medium has made the postbiotic RS5 more feasible and cost-effective to be adopted as a feed supplement for various livestock industries.

Supplementation of postbiotic RI11 improves antioxidant enzyme activity, upregulated gut barrier genes, and reduced cytokine, acute phase protein, and heat shock protein 70 gene expression levels in heat-stressed broilers.

The aim of this work was to evaluate the impacts of feeding different levels of postbiotic RI11 on antioxidant enzyme activity, physiological stress indicators, and cytokine and gut barrier gene expression in broilers under heat stress. A total of 252 male broilers Cobb 500 were allocated in cages in environmentally controlled chambers. All the broilers received the same basal diet from 1 to 21 d. On day 22, the broilers were weighed and grouped into 7 treatment groups and exhibited to cyclic high temperature at 36 ± 1°C for 3 h per day until the end of the experiment. From day 22 to 42, broilers were fed with one of the 7 following diets: negative control, basal diet (0.0% RI11) (NC group); positive control, NC diet + 0.02% (w/w) oxytetracycline (OTC group); antioxidant control, NC diet + 0.02% (w/w) ascorbic acid. The other 4 other groups were as follows: NC diet + 0.2% cell-free supernatant (postbiotic RI11) (v/w), NC diet + 0.4% cell-free supernatant (postbiotic RI11) (v/w), NC diet + 0.6% cell-free supernatant (postbiotic RI11) (v/w), and NC diet + 0.8% cell-free supernatant (postbiotic RI11) (v/w). Supplementation of different levels (0.4, 0.6, and 0.8%) of postbiotic RI11 increased plasma glutathione peroxidase, catalase, and glutathione enzyme activity. Postbiotic RI11 groups particularly at levels of 0.4 and 0.6% upregulated the mRNA expression of IL-10 and downregulated the IL-8, tumor necrosis factor alpha, heat shock protein 70, and alpha-1-acid glycoprotein levels compared with the NC and OTC groups. Feeding postbiotic RI11, particularly at the level of 0.6%, upregulated ileum zonula occludens-1 and mucin 2 mRNA expressions. However, no difference was observed in ileum claudin 1, ceruloplasmin, IL-6, IL-2, and interferon expression, but downregulation of occludin expression was observed as compared with the NC group. Supplementation of postbiotic RI11 at different levels quadratically increased plasma glutathione peroxidase, catalase and glutathione, IL-10, mucin 2, and zonula occludens-1 mRNA expression and reduced plasma IL-8, tumor necrosis factor alpha, alpha-1-acid glycoprotein, and heat shock protein 70 mRNA expression. The results suggested that postbiotics produced from Lactiplantibacillus plantarum RI11 especially at the level of 0.6% (v/w) could be used as an alternative to antibiotics and natural sources of antioxidants in poultry feeding.

Growth Performance, Cytokine Expression, and Immune Responses of Broiler Chickens Fed a Dietary Palm Oil and Sunflower Oil Blend Supplemented With L-Arginine and Varying Concentrations of Vitamin E.

This study set out to examine the combined effects of the supplementation of a dietary palm oil (PO) and sunflower oil (SO) blend, 0. 25% L-Arginine (L-Arg), and different levels of vitamin E (Vit E) on growth performance, fat deposition, cytokine expression, and immune response in broilers. A total of 216 1-day-old male broiler chicks (Cobb500) were randomly distributed into six dietary groups as follows: Diet 1: 6% palm oil (negative control); Diet 2: PO and SO blend (4% palm oil and 2% sunflower oil) + 0.25% L-Arg (positive control); Diet 3: (PO and SO blend + 0.25% L-Arg) + 20 mg/kg Vit E; Diet 4: (PO and SO blend + 0.25% L-Arg) + 50 mg/kg Vit E; Diet 5: (PO and SO blend + 0.25% L-Arg) + 100 mg/kg Vit E; and Diet 6: (PO and SO blend + 0.25% L-Arg) + 150 mg/kg Vit E. Weight gain and serum IgG and IgM increased while feed conversion ratio, fat deposition, and plasma cholesterol decreased in broilers fed Vit E with the oil blend and L-Arg, compared to those fed the negative control (Diet 1). Expression of IFN and TNF-α were reduced, whereas TGF-ß1 was up-regulated as the level of Vit E increased in the broiler diets. In summary, the combination of oil blend, L-Arg, and Vit E at a level of 50 mg/kg increased the performance and altered the expression of cytokines that may positively influence immune function in broiler chickens.

Dietary Supplementation of Postbiotics Mitigates Adverse Impacts of Heat Stress on Antioxidant Enzyme Activity, Total Antioxidant, Lipid Peroxidation, Physiological Stress Indicators, Lipid Profile and Meat Quality in Broilers.

The purpose of this work was to evaluate the impacts of feeding different postbiotics on oxidative stress markers, physiological stress indicators, lipid profile and meat quality in heat-stressed broilers. A total of 252 male Cobb 500 (22-day-old) were fed with 1 of 6 diets: A basal diet without any supplementation as negative control (NC); basal diet + 0.02% oxytetracycline served as positive control (PC); basal diet + 0.02% ascorbic acid (AA); or the basal diet diet + 0.3% of RI11, RS5 or UL4 postbiotics. Postbiotics supplementation, especially RI11 increased plasma activity of total-antioxidant capacity (T-AOC), catalase (CAT) and glutathione (GSH), and decreased alpha-1-acid-glycoprotein (α1-AGP) and ceruloplasmin (CPN) compared to NC and PC groups. Meat malondialdehyde (MDA) was lower in the postbiotic groups than the NC, PC and AA groups. Plasma corticosterone, heat shock protein70 (HSP70) and high density lipoprotein (HDL) were not affected by dietary treatments. Postbiotics decreased plasma cholesterol concentration compared to other groups, and plasma triglyceride and very low density lipoprotein (VLDL) compared to the NC group. Postbiotics increased breast meat pH, and decreased shear force and lightness (L*) compared to NC and PC groups. The drip loss, cooking loss and yellowness (b*) were lower in postbiotics groups compared to other groups. In conclusion, postbiotics particularly RI11 could be used as an alternative to antibiotics and natural sources of antioxidants for heat-stressed broilers.

Effects of Feeding Different Postbiotics Produced by Lactobacillus plantarum on Growth Performance, Carcass Yield, Intestinal Morphology, Gut Microbiota Composition, Immune Status, and Growth Gene Expression in Broilers under Heat Stress.

The effects of feeding different postbiotics on growth performance, carcass yield, intestinal morphology, gut microbiota, immune status, and growth hormone receptor (GHR) and insulin-like growth factor 1 (IGF-1) gene expression in broilers under heat stress were assessed in this study. A total of 252 one-day-old male broiler chicks (Cobb 500) were randomly assigned in cages in identical environmentally controlled chambers. During the starter period from 1 to 21 days, all the birds were fed the same basal diet. On day 22, the birds were weighed and randomly divided into six treatment groups and exposed to cyclic high temperature at 36 ± 1 °C for 3 h per day from 11:00 to 14:00 until the end of the experiment. From day 22 to 42 (finisher period), an equal number of birds were subjected to one of the following diets: NC (negative control) basal diet; PC (positive control) basal diet + 0.02% oxytetracycline; or AA (ascorbic acid) basal diet + 0.02% ascorbic acid. The other three groups (RI11, RS5 and UL4) were basal diet + 0.3% different postbiotics (produced from different Lactobacillus plantarum strains, and defined as RI11, RS5 and UL4, respectively). The results demonstrated that birds fed RI11 diets had significantly higher final body weight, total weight gain and average daily gain than the birds that received the NC, PC and AA treatments. The feed conversion ratio was significantly higher in the RI11 group compared with the other groups. Carcass parameters were not affected by the postbiotic-supplemented diet. Postbiotic supplementation improved villi height significantly in the duodenum, jejunum and ileum compared to the NC, PC and AA treatments. The crypt depth of the duodenum and ileum was significantly higher in NC group compared to other treatment groups except RI11 in duodenum, and UL4 in ileum was not different with NC groups. The villus height to crypt depth ratio of duodenum and ileum was significantly higher for the postbiotic treatment groups and AA than the PC and NC treatment groups. The postbiotic RI11 group recorded significantly higher caecum total bacteria and Lactobacillus count and lower Salmonella count compared to the NC and PC treatment groups. The Bifidobacterium population in the NC group was significantly lower compared to the other treatment groups. The postbiotic (RI11, RS5 and UL4) and AA treatment groups showed lower Enterobacteriaceae and E. coli counts and caecal pH than the NC and PC treatment groups. The plasma immunoglobulin M (IgM) level was significantly higher in the birds receiving postbiotic RI11 than those receiving other treatments. The plasma immunoglobulin G (IgG) level was higher in the RI11 treatment group than in the NC, AA and RS5 groups. The plasma immunoglobulin A (IgA) level was not affected by postbiotic supplements. The hepatic GHR mRNA expression level was significantly increased in birds fed postbiotics RI11, RS5 and UL4, AA and PC compared to the NC-fed birds. Postbiotic RI11 led to significantly higher hepatic IGF-1 mRNA expression level compared to the NC, PC, and AA treatments. Mortality was numerically lesser in the postbiotic treatment groups, but not significantly different among all the treatments. In conclusion, among the postbiotics applied in the current study as compared with NC, PC and AA, RI11 could be used as a potential alternative antibiotic growth promoter and anti-stress treatment in the poultry industry.

Effects of postbiotic supplementation on growth performance, ruminal fermentation and microbial profile, blood metabolite and GHR, IGF-1 and MCT-1 gene expression in post-weaning lambs.

BACKGROUND: Postbiotics have been established as potential feed additive to be used in monogastric such as poultry and swine to enhance health and growth performance. However, information on the postbiotics as feed additive in ruminants is very limited. The aim of this study was to elucidate the effects of supplementation of postbiotics in newly-weaned lambs on growth performance, digestibility, rumen fermentation characteristics and microbial population, blood metabolite and expression of genes related to growth and volatile fatty acid transport across the rumen epithelium. RESULTS: Postbiotic supplementation increased weight gain, feed intake, nutrient intake and nutrient digestibility of the lambs. No effect on ruminal pH and total VFA, whereas butyrate and ruminal ammonia-N concentration were improved. The lambs fed with postbiotics had higher blood total protein, urea nitrogen and glucose. However, no difference was observed in blood triglycerides and cholesterol levels. Postbiotics increased the population of fibre degrading bacteria but decreased total protozoa and methanogens in rumen. Postbiotics increased the mRNA expression of hepatic IGF-1 and ruminal MCT-1. CONCLUSIONS: The inclusion of postbiotics from L. plantarum RG14 in newly-weaned lambs improved growth performance, nutrient intake and nutrient digestibility reflected from better rumen fermentation and microbial parameters, blood metabolites and upregulation of growth and nutrient intake genes in the post-weaning lambs.

Postbiotic metabolites produced by Lactobacillus plantarum strains exert selective cytotoxicity effects on cancer cells.

BACKGROUND: Lactobacillus plantarum, a major species of Lactic Acid Bacteria (LAB), are capable of producing postbiotic metabolites (PM) with prominent probiotic effects that have been documented extensively for rats, poultry and pigs. Despite the emerging evidence of anticancer properties of LAB, very limited information is available on cytotoxic and antiproliferative activity of PM produced by L. plantarum. Therefore, the cytotoxicity of PM produced by six strains of L. plantarum on various cancer and normal cells are yet to be evaluated. METHODS: Postbiotic metabolites (PM) produced by six strains of L. plantarum were determined for their antiproliferative and cytotoxic effects on normal human primary cells, breast, colorectal, cervical, liver and leukemia cancer cell lines via MTT assay, trypan blue exclusion method and BrdU assay. The toxicity of PM was determined for human and various animal red blood cells via haemolytic assay. The cytotoxicity mode was subsequently determined for selected UL4 PM on MCF-7 cells due to its pronounced cytotoxic effect by fluorescent microscopic observation using AO/PI dye reagents and flow cytometric analyses. RESULTS: UL4 PM exhibited the lowest IC50 value on MCF-7, RG14 PM on HT29 and RG11 and RI11 PM on HL60 cell lines, respectively from MTT assay. Moreover, all tested PM did not cause haemolysis of human, dog, rabbit and chicken red blood cells and demonstrated no cytotoxicity on normal breast MCF-10A cells and primary cultured cells including human peripheral blood mononuclear cells, mice splenocytes and thymocytes. Antiproliferation of MCF-7 and HT-29 cells was potently induced by UL4 and RG 14 PM respectively after 72 h of incubation at the concentration of 30% (v/v). Fluorescent microscopic observation and flow cytometric analyses showed that the pronounced cytotoxic effect of UL4 PM on MCF-7 cells was mediated through apoptosis. CONCLUSION: In conclusion, PM produced by the six strains of L. plantarum exhibited selective cytotoxic via antiproliferative effect and induction of apoptosis against malignant cancer cells in a strain-specific and cancer cell type-specific manner whilst sparing the normal cells. This reveals the vast potentials of PM from L. plantarum as functional supplement and as an adjunctive treatment for cancer.

Fatty acid composition, fat deposition, lipogenic gene expression and performance of broiler fed diet supplemented with different sources of oil.

The present study assessed the effect of feeding palm oil (PO), sunflower oil (SO) and their combination on performance, fat deposition, fatty acid composition and lipogenic gene expression of broilers reared for 42 days. A total of 144 1-day-old broilers (Cobb500) were randomly allotted into four treatment diets with each having six replicates of six chicks in each replicate following a completely randomized design. Live weight gain and feed efficiency was significantly (P < 0.05) higher in birds fed with a combination of oil sources compared to controls. Birds fed with the combination of oil and SO alone had higher carcass yield and lower abdominal fat. Higher (P < 0.05) concentrations of unsaturated fatty acids (UFA) and lower concentrations of palmitic acid and saturated fatty acid (SFA) was found in birds fed SO alone and combinations of SO and PO. Furthermore, the outcomes showed that birds fed diet supplemented with SO and the combination of SO and PO down-regulated gene expression of key hepatic lipogenic enzymes of fatty acids synthase (FAS), acetyl-CoA carboxylase (ACC) and stearoyl-CoA desaturase (SCD). These findings suggest that the diet containing the combination of 2% PO and 4% SO may reduce hepatic lipogenesis, as well as lower abdominal fat content of broilers.

Effects of dietary postbiotic and inulin on growth performance, IGF1 and GHR mRNA expression, faecal microbiota and volatile fatty acids in broilers.

BACKGROUND: Postbiotics (metabolic products by lactic acid bacteria) and prebiotics have been established as substitute to antibiotics in order to enhance immunity and growth performance in broiler chickens. Nonetheless, insufficient information is available on the effects of postbiotics and prebiotics combination on growth performance, faecal microbiota, pH and volatile fatty acids (VFA), as well as liver insulin like growth factor 1 (IGF1) and growth hormone receptor (GHR) mRNA expressions in broiler chickens. The aim of this experiment was to evaluate the effects of different types of postbiotics with different levels of prebiotic (inulin) on broiler for those parameters. RESULTS: The results showed that birds fed T3: (0.3 % RI11 + 0.8 % Inulin), T4: (0.3 % RI11 + 1.0 % Inulin), and T6: (0.3 % RG14+ 1.0 % Inulin) had higher (p < 0.05) final body weight (BW) and total weight gain (WG) than other treatments. Birds fed T3 had lower feed conversion ratio (FCR) which was significantly different from those fed with negative control diet but was similar to other treatments. Postbiotic and inulin increased (p < 0.05) faecal lactic acid bacteria (LAB) and reduced (p < 0.05) Enterobacteriaceae count. Birds fed T4 and T6 had higher faecal acetic acid and propionic acid respectively, and both had higher total VFA and lactic acid bacteria but lower pH and Enterobacteriaceae (ENT) counts compared to other treatments. The liver of birds fed T4 and T6 had higher IGF1 expression compared to other treatments while T6 had higher GHR mRNA expression compared to other treatments. CONCLUSIONS: Results indicate that the addition of postbiotics and inulin combinations had beneficial effects on total BW, feed efficiency, mucosa architecture and IGF1 and GHR mRNA expression in broiler chickens.

Effects of feeding different postbiotic metabolite combinations produced by Lactobacillus plantarum strains on egg quality and production performance, faecal parameters and plasma cholesterol in laying hens.

BACKGROUND: Probiotics are beneficial bacteria that are able to colonize the host digestive system, increasing the natural flora and preventing colonization of pathogenic organisms and thus, securing optimal utility of the feed. However, commercial probiotic often do not meet the expected standards and the viability of the efficacy of these strains remains questionable. Another major issue has been highlighted in relation to the application of antibiotic resistant probiotics, the antibiotic resistant gene can be transferred between organisms. Recently, postbiotic metabolites produced from microbes have been extensively studied as feed additive in order to substitute in-feed antibiotics. RESULTS: No significant difference (P > 0.05) was found among the treatment groups on overall feed intake, egg weight, egg mass and feed conversion efficiency. COM456 had a significant reduction (P < 0.05) in faecal pH compared to the other groups at 28 weeks of age onwards. COM456 had significant higher (P < 0.05) level of lactic acid bacteria counts from 30 weeks of age onwards, followed by COM246 and COM345 at 32 and 34 weeks of age, respectively. Significant reduction of faecal Enterobacteriaceae (P < 0.05) were observed in COM246 and COM456 from 30 weeks of age onwards. The lowest levels (P < 0.05) of plasma and egg yolk cholesterol were observed in COM456, followed by COM345 and COM246. There was no significant difference in terms of yolk weight between the treatment groups. Significant higher (P < 0.05) content of C18:3, C20:2 and C22:6 were found in treatments supplemented with metabolite combinations as compared with the control group. CONCLUSIONS: The present study demonstrated the positive effects of metabolite combinations supplementation in laying hens. Increase in hen-day egg production was observed in all treatments supplemented with metabolite combinations. In addition, the metabolite combinations, COM456 had reduced the faecal pH and faecal Enterobacteriaceae population, improved the faecal lactic acid bacteria, reduced the plasma and yolk cholesterol and improved the faecal volatile fatty acids content. Postbiotic metabolite combinations can be used as an alternative feed additive to achieve high productivity and better animal health while reducing the use of conventional chemotherapeutic agents such as in-feed antimicrobials.