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Fulltext 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

Khatun J, Loh TC, Foo HL, Akit H, Khan KI

Front Vet Sci, 2020;7:619.
PMID: 33195499 DOI: 10.3389/fvets.2020.00619

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.
Fulltext Comparative Efficacy of Selected Phytobiotics with Halquinol and Tetracycline on Gut Morphology, Ileal Digestibility, Cecal Microbiota Composition and Growth Performance in Broiler Chickens

Basit MA, Kadir AA, Loh TC, Abdul Aziz S, Salleh A, Zakaria ZA, et al.

Animals (Basel), 2020 Nov 19;10(11).
PMID: 33227911 DOI: 10.3390/ani10112150

The current experiment was designed to estimate the comparative efficacy of selected phytobiotics Persicaria odorata leaf meal (POLM) and Piper betle leaf meal (PBLM) with halquinol, and tetracycline in broiler chickens. The 150-day-old broiler chickens were randomly assigned to five dietary groups. The dietary supplementation groups were the basal diet (BD), which served as the negative control (NC), and BD + 0.2 g/kg tetracycline, which served as the positive control (PC); BD + 0.03 g/kg halquinol (HAL), BD + 8 g/kg POLM (Po8), and BD + 4 g/kg PBLM (Pb4) were the treatment groups. Growth performance, gut morphology, ileal digestibility, and cecal microbiota composition were measured. On day 21, the body weight gain (BWG) was enhanced (p < 0.05) in the broiler chickens fed on phytobiotics (Po8 and Pb4) relative to the NC group, however, on day 42 and in terms of overall growth performance, BWG was enhanced (p < 0.05 in diets (Po8, Pb4, HAL and PC) in comparison with the NC group. Conversely, feed conversion ratio (FCR) was recorded reduced (p < 0.05) in Pb4, Po8, HAL, and PC group in comparison with the NC group. Supplementation of phytobiotics (Po8 and Pb4), HAL and PC, positively improved the gut morphology compared to the NC group. Furthermore, the maximum (p < 0.05) villus height (VH) in duodenum and jejunum was observed in broilers fed on diet Pb4. Supplementation of phytobiotics, HAL and PC, improved (p < 0.05) the digestibility of dry matter (DM) (except for HAL), organic matter (OM), crude protein (CP), ether extract (EE), and ash compared to the NC group. Dietary supplementation of phytobiotics (Po8 and Pb4), HAL and PC, significantly reduced the E. coli, Salmonella, and Staphylococcus aureus (except for HAL) counts compared to the NC group. However, supplementation of Pb4 resulted in significantly decreased total anaerobic bacteria and Clostridium spp. counts compared to the NC group. In addition, supplementation of phytobiotics significantly increased the Lactobacillus count compared to HAL, PC, and NC groups. In conclusion, dietary supplementation of phytobiotics improved the gut morphology, positively modulated and maintained the dynamics of cecal microbiota with enhanced nutrient digestibility, thus, increased the growth performance. Based on current results, phytobiotics could be used as an alternative to AGPs for sustainable broiler chicken production.
Fulltext Effects of dietary postbiotic and inulin on growth performance, IGF1 and GHR mRNA expression, faecal microbiota and volatile fatty acids in broilers

Kareem KY, Loh TC, Foo HL, Akit H, Samsudin AA

BMC Vet Res, 2016;12(1):163.
PMID: 27496016 DOI: 10.1186/s12917-016-0790-9

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

Ooi MF, Foo HL, Loh TC, Mohamad R, Rahim RA, Ariff A

Sci Rep, 2021 Apr 07;11(1):7617.
PMID: 33828119 DOI: 10.1038/s41598-021-87081-6

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.
Fulltext 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

Humam AM, Loh TC, Foo HL, Izuddin WI, Zulkifli I, Samsudin AA, et al.

Poult Sci, 2021 Mar;100(3):100908.
PMID: 33518339 DOI: 10.1016/j.psj.2020.12.011

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.
Fulltext Comparative Studies of Inhibitory and Antioxidant Activities, and Organic Acids Compositions of Postbiotics Produced by Probiotic Lactiplantibacillus plantarum Strains Isolated From Malaysian Foods

Chang HM, Foo HL, Loh TC, Lim ETC, Abdul Mutalib NE

Front Vet Sci, 2020;7:602280.
PMID: 33575277 DOI: 10.3389/fvets.2020.602280

Despite inflammation being a protective natural defense against imbalance stressors in the body, chronic inflammation could lead to the deterioration of immune response, low production, and poor performance in livestock as well as severe economic losses to the farmers. Postbiotics produced by Lactiplantibacillus plantarum has been reported recently to be a natural source of antioxidant, promoting growth performance, anti-inflammation, and immune responses. However, the effects of fermentation media on the compositions of L. plantarum postbiotic have not been reported elsewhere. Hence, a comparative study was conducted to compare the volatile compounds, organic acid composition, and antioxidant and antimicrobial activities of postbiotics produced by six strains of L. plantarum cultivated by using formulated media and the commercial de Man, Rogosa, and Sharpe (MRS) medium as a control. Postbiotics RG14, RI11, and UL4 produced by using formulated media exhibited higher inhibitory activity against Pediococcus acidilactici 446, Escherichia coli E-30, Salmonella enterica CS3, and vancomycin-resistant Enterococci except for Listeria monocytogenes LS55. As for the antioxidant activity, hydroxyl radical scavenging activity was enhanced in formulated media, whereas reducing power activity was the highest in postbiotic RI11. Three organic acids, namely, acetic acid, caproic acid, and lactic acid, were detected in the postbiotic produced by various L. plantarum strains. The concentration of acetic acid was influenced by the fermentation media, whereas caproic acid was detected as the highest in postbiotic RG11. Lactic acid was the predominant compound detected in all the postbiotics and had the significantly highest concentration in postbiotic RS5 when produced by using the MRS medium. Intermediary and pyrrole compounds were the other main compounds that were detected by using GC-MS. Positive correlations were found between organic acid production and inhibitory activity, as well as antioxidant activity exhibited by postbiotics. In conclusion, the compositions and functional characteristics of postbiotics produced by the six strains of L. plantarum were strain-dependent and affected greatly by the fermentation medium. The effects of postbiotic composition on the functional characteristics of postbiotics were elucidated in this study to warrant their applications as a promising beneficial natural growth promoter for the livestock industry.
Fulltext Meat Quality, Fatty Acid Content and NMR Metabolic Profile of Dorper Sheep Supplemented with Bypass Fats

Behan AA, Akhtar MT, Loh TC, Fakurazi S, Kaka U, Muhamad A, et al.

Foods, 2021 May 19;10(5).
PMID: 34069633 DOI: 10.3390/foods10051133

The supplementation of rumen bypass fat (RBF) has remained one of the preferred approaches used to decrease undesirable saturated fatty acids (FA) and increase beneficial unsaturated FA in the meat. This study was planned to evaluate the influences of rumen bypass fats on meat quality, fatty acid and metabolic profiles in male Dorper sheep (n = 36) with 24.66 ± 0.76 kg (mean ± standard error) initial body weight. Treatment comprised a basal diet (30:70 rice straw to concentrate) with no added RBF as a control (CON), basal diet with prilled fat (PF), basal diet with prilled fat plus lecithin (PFL) and basal diet with calcium soap of palm fatty acids (CaS). The findings revealed that cooking loss, drip loss and shear force in longissimus dorsi (LD) muscle were not affected by RBF supplementation, while meat pH was significantly higher in the CaS on aging day 1. However, the diet supplemented with prilled fat and lecithin modified the meat's fatty acid profile significantly by increasing unsaturated fatty acids and decreasing saturated fats. The relative quantification of the major differentiating metabolites found in LD muscle of sheep showed that total cholesterol, esterified cholesterol, choline, glycerophosphocholine and glycerophospholipids were significantly lower in CaS and PFL diets, while glycerol and sphingomyelin were significantly higher in CaS and PFL diets. Most of the metabolites in the liver did not show any significant difference. Based on our results, the supplementation of protected fats did not have a negative influence on meat quality and the meat from Dorper sheep fed prilled fat with lecithin contained more healthy fatty acids compared to other diets.
Fulltext Biodegradation of palm kernel cake by cellulolytic and hemicellulolytic bacterial cultures through solid state fermentation

Alshelmani MI, Loh TC, Foo HL, Lau WH, Sazili AQ

ScientificWorldJournal, 2014;2014:729852.
PMID: 25019097 DOI: 10.1155/2014/729852

Four cellulolytic and hemicellulolytic bacterial cultures were purchased from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ) and the American Type Culture Collection (ATCC). Two experiments were conducted; the objective of the first experiment was to determine the optimum time period required for solid state fermentation (SSF) of palm kernel cake (PKC), whereas the objective of the second experiment was to investigate the effect of combinations of these cellulolytic and hemicellulolytic bacteria on the nutritive quality of the PKC. In the first experiment, the SSF was lasted for 12 days with inoculum size of 10% (v/w) on different PKC to moisture ratios. In the second experiment, fifteen combinations were created among the four microbes with one untreated PKC as a control. The SSF lasted for 9 days, and the samples were autoclaved, dried, and analyzed for proximate analysis. Results showed that bacterial cultures produced high enzymes activities at the 4th day of SSF, whereas their abilities to produce enzymes tended to be decreased to reach zero at the 8th day of SSF. Findings in the second experiment showed that hemicellulose and cellulose was significantly (P < 0.05) decreased, whereas the amount of reducing sugars were significantly (P < 0.05) increased in the fermented PKC (FPKC) compared with untreated PKC.
Fulltext 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

Loh TC, Choe DW, Foo HL, Sazili AQ, Bejo MH

BMC Vet Res, 2014;10:149.
PMID: 24996258 DOI: 10.1186/1746-6148-10-149

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.
Effects of feeding fermented fish on egg cholesterol content in hens

Loh TC, Law FL, Goh YM, Foo HL, Zulkifli I

Anim Sci J, 2009 Feb;80(1):27-33.
PMID: 20163464 DOI: 10.1111/j.1740-0929.2008.00591.x

This study was conducted to investigate the effects of feeding fermented fish (FF) to layers on laying performance, and polyunsaturated fatty acid and cholesterol levels in eggs and plasma. A total of 96, 13-week-old Babcock B380 pullets were used in this study. They were randomly assigned to four numerically equal groups with eight replicates per treatment, three birds per replicate. All the birds were housed in individual cages. The dietary treatments were: Control diet, without FF; FF3 diet containing 3% (w/w) FF, FF6 diet containing 6% (w/w) FF and FF9 diet containing 9% (w/w) FF. The study was carried out for 16 weeks inclusive of two weeks of adjustment. Weekly feed intake and egg production were recorded. Blood plasma cholesterol and fatty acid profiles were assayed at the end of the experiment. FF did not enhance (P > 0.05) egg mass but (P < 0.05) decreased egg weight slightly. However, egg yolk cholesterol and plasma cholesterol concentrations were reduced (P < 0.05) by FF. The n-6:n-3 fatty acids ratio in the egg yolk (Control = 7.9, FF9 = 6.2) and plasma (Control = 10.6, FF9 = 6.2) were decreased by feeding FF. Moreover, FF was able to increase (P < 0.05) the docosahexaenoic acid (DHA) concentrations in egg yolk and plasma. In conclusion, this study demonstrated that FF increased DHA and reduced egg yolk cholesterol in poultry eggs.
Fulltext Blood lipid profiles, fatty acid deposition and expression of hepatic lipid and lipoprotein metabolism genes in laying hens fed palm oils, palm kernel oil, and soybean oil

Izuddin WI, Loh TC, Nayan N, Akit H, Noor AM, Foo HL

Front Vet Sci, 2023;10:1192841.
PMID: 37519991 DOI: 10.3389/fvets.2023.1192841

The palm oil, palm kernel oil and soybean oil have unique and distinctive fatty acid chain length and saturation profiles, and how they affect lipid peroxidation, fatty acid intake and metabolism is worth exploring in poultry. This study elucidated the influence the dietary oils on lipid peroxidation, blood lipid profiles, fatty acid deposition of liver, serum and yolk and the expression of liver genes related to lipid and lipoprotein metabolism in laying hens. About 150 Hisex brown laying hens were fed diets containing crude palm oil (CPO), red palm oil (RPO), refined palm oil (RBD), palm kernel oil (PKO) or soybean oil (SBO) for 16 weeks. Serum, liver and yolk lipid peroxidation were not different between dietary oils. The PKO increased liver, serum and yolk medium-chain fatty acids (MCFA). There was no difference in liver saturated fatty acids (SFA). The CPO and RPO reduced serum SFA, but the PKO increased yolk SFA. The SBO increased polyunsaturated fatty acids (PUFA) in liver serum and yolk. No difference in liver elaidic acid (C18:1-trans), but SBO lowered elaidic acid (C18:1-trans) in serum. Higher very-low density lipoprotein (VLDL) in CPO than RPO and SBO and greater serum lipase in CPO, RBD and PKO than SBO. There was no difference in sterol regulatory element-binding protein 2 (SREBP-II) between oils. Apolipoprotein VLDL-II (APOVLDL2) was upregulated in palm oils and apolipoprotein B-100 (APOB) in RBD. Downregulation in peroxisome proliferator-activated receptor-alpha (PPAR-α), peroxisome proliferator-activated receptor gamma (PPAR-γ) and low-density lipoprotein receptor (LDLR) was observed in palm oils and PKO. In conclusion, different dietary oils greatly influence several aspects of fatty acid metabolism, deposition and lipoprotein profiles but have no influence on reducing lipid peroxidation.
Fulltext 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

Azizi MN, Loh TC, Foo HL, Akit H, Izuddin WI, Yohanna D

Animals (Basel), 2023 May 09;13(10).
PMID: 37238013 DOI: 10.3390/ani13101582

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.
Fulltext Prevalence of antibiotic resistance in lactic acid bacteria isolated from the faeces of broiler chicken in Malaysia

Shazali N, Foo HL, Loh TC, Choe DW, Abdul Rahim R

Gut Pathog, 2014 Jan 22;6(1):1.
PMID: 24447766 DOI: 10.1186/1757-4749-6-1

BACKGROUND: Probiotics are commonly used as feed additive to substitute antibiotic as growth promoter in animal farming. Probiotic consists of lactic acid bacteria (LAB), which enhance the growth and health of the animal. Probiotic also have higher possibility to become pathogenic bacteria that may carry antibiotic resistant gene that can be transmitted to other LAB species. The aim of this study was to identify the LAB species in the faeces of broiler chicken and to determine the prevalence of antibiotic resistant in LAB of broiler chicken.
METHODS: Sixty faeces samples were collected from wet markets located in Klang Valley of Malaysia for the isolation of LAB using de-Mann Rogosa Sharpe medium. Thirteen species of LAB were obtained in this study and the identification of LAB was performed by using API test kit on the basis of carbohydrate fermentation profile. Antibiotic susceptibility assay was then carried out to determine the prevalence of LAB antibiotic resistance.
RESULTS: Lactococcus lactis subsp lactis was found in nine out of sixty faecal samples. Lactobacillus paracasei was the second common LAB species isolated from chicken faecal. No significant difference (P > 0.05) was found between the occurrence of Lactobacillus brevis, Lactobacillus curvatus, Lactobacillus plantarum, Leuconostoc lactis mesenteroides subsp mesenteroides/dectranium and Pediococcus pentosaceus isolated from 5 different locations. Most of the isolated LAB was resistant to antibiotic and high variability of the antibiotic resistance was observed among the LAB against 15 types of antibiotics. Penicillin, amoxicillin, chloramphenicol, and ampicillin had significant higher (P< 0.05) inhibitory zone than nalidixic acid, gentamycin, sulphamethoxazole, kanamycin, and streptomycin.
CONCLUSIONS: Many species of LAB were isolated from the faecal samples of broiler chicken that resistance to the common antibiotics used in the farm. The development of resistant against antibiotics in LAB can be attributed to the long term exposure of antibiotic as growth promoter and therapeutic agents. Thus, it is essential to advise farmer the safety measure of antibiotic application in animal farming. Additionally, the supplementation of probiotic in animal feeding also needs more attention and close monitoring.
Fulltext Characterization of cellulolytic bacterial cultures grown in different substrates

Alshelmani MI, Loh TC, Foo HL, Lau WH, Sazili AQ

ScientificWorldJournal, 2013;2013:689235.
PMID: 24319380 DOI: 10.1155/2013/689235

Nine aerobic cellulolytic bacterial cultures were obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ) and the American Type Culture Collection (ATCC). The objectives of this study were to characterize the cellulolytic bacteria and to determine the optimum moisture ratio required for solid state fermentation (SSF) of palm kernel cake (PKC). The bacteria cultures were grown on reconstituted nutrient broth, incubated at 30°C and agitated at 200 rpm. Carboxymethyl cellulase, xylanase, and mannanase activities were determined using different substrates and after SSF of PKC. The SSF was conducted for 4 and 7 days with inoculum size of 10% (v/w) on different PKC concentration-to-moisture ratios: 1 : 0.2, 1 : 0.3, 1 : 0.4, and 1 : 0.5. Results showed that Bacillus amyloliquefaciens 1067 DSMZ, Bacillus megaterium 9885 ATCC, Paenibacillus curdlanolyticus 10248 DSMZ, and Paenibacillus polymyxa 842 ATCC produced higher enzyme activities as compared to other bacterial cultures grown on different substrates. The cultures mentioned above also produced higher enzyme activities when they were incubated under SSF using PKC as a substrate in different PKC-to-moisture ratios after 4 days of incubation, indicating that these cellulolytic bacteria can be used to degrade and improve the nutrient quality of PKC.
Effects of Postbiotics and Paraprobiotics as Replacements for Antibiotics on Growth Performance, Carcass Characteristics, Small Intestine Histomorphology, Immune Status and Hepatic Growth Gene Expression in Broiler Chickens

Danladi Y, Loh TC, Foo HL, Akit H, Md Tamrin NA, Naeem Azizi M

Animals (Basel), 2022 Apr 03;12(7).
PMID: 35405905 DOI: 10.3390/ani12070917

Background: This experiment was designed to investigate how replacing antibiotics with postbiotics and paraprobiotics could affect growth performance, small intestine morphology, immune status, and hepatic growth gene expression in broiler chickens. Methods: The experiment followed a completely randomized design (CRD) in which eight treatments were replicated six times with seven birds per replicate. A total of 336, one-day-old (COBB 500) chicks were fed with the eight treatment diets, which include T1 = negative control (Basal diet), T2 = positive control (Basal diet + 0.01% (w/w) Oxytetracycline), T3 = Basal diet + 0.2% (v/w) postbiotic TL1, T4 = Basal diet + 0.2% (v/w) postbiotic RS5, T5 = Basal diet + 0.2% (v/w) paraprobiotic RG11, T6 = Basal diet + 0.2% (v/w) postbiotic RI11, T7 = Basal diet + 0.2% (v/w) paraprobiotic RG14, T8 = Basal diet + 0.2% (v/w) paraprobiotic RI11, for 35 days in a closed house system. Results: The growth performance indicators (final body weight, cumulative weight gain, and feed conversion ratio) were not significantly (p > 0.05) affected by the dietary treatments. However, feed intake recorded a significant (p < 0.05) change in the starter and finisher phases across the dietary treatments. Paraprobiotic RG14 had significantly (p < 0.05) lower abdominal fat and intestines. Villi heights were significantly (p < 0.05) increased, while the crypt depth decreased significantly due to dietary treatments. The dietary treatments significantly influenced colon mucosa sIgA (p < 0.05). Similarly, plasma immunoglobulin IgM level recorded significant (p < 0.05) changes at the finisher phase. In this current study, the hepatic GHR and IGF-1 expressions were significantly (p < 0.05) increased by postbiotics and paraprobiotics supplementation. Conclusions: Therefore, it was concluded that postbiotics and paraprobiotics differ in their effect on broiler chickens. However, they can replace antibiotics without compromising the growth performance, carcass yield, and immune status of broiler chickens.
Fulltext Palm kernel cake and decanter cake as broiler feed ingredients: Effects on immunity, gut microbiota, antioxidant gene expression and plasma lipid profile

Azizi MN, Loh TC, Chung ELT, Aziz MFA, Foo HL

Poult Sci, 2026 Oct 05;105(12):107902.
PMID: 42854500 DOI: 10.1016/j.psj.2026.107902

The present study examined the impact of feeding broilers diets containing blended palm kernel cake (PKC) and decanter cake (DC) on immune response, cecal microbiota, antioxidant gene expression, and plasma lipid profile in broiler chickens. A total of 1,260 male Ross 308 broiler chicks were reared for 35 days in a completely randomized design (CRD). Birds were assigned to 18 dietary treatments, with seven replicates of 10 birds per treatment. These treatments comprised a basal negative control, single-ingredient positive controls (5% PKC or DC), and PKC:DC combinations at 30:70, 50:50, and 70:30 ratios with inclusion levels of 2-10%. Dietary supplementation with PKC and DC blends significantly enhanced (p  0.05). Dietary treatments did not influence (p > 0.05) the concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), or interleukin-6 (IL-6). In contrast, interleukin-10 (IL-10) expression was upregulated (p  0.05). Overall, moderate inclusion levels of PKC and DC blends improved IL-10 expression, cecal Lactobacillus populations, and GPX1 gene expression without altering the measured plasma lipid concentrations.
Fulltext Postbiotic metabolites produced by Lactobacillus plantarum strains exert selective cytotoxicity effects on cancer cells

Chuah LO, Foo HL, Loh TC, Mohammed Alitheen NB, Yeap SK, Abdul Mutalib NE, et al.

BMC Complement Altern Med, 2019 Jun 03;19(1):114.
PMID: 31159791 DOI: 10.1186/s12906-019-2528-2

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.
Fulltext 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

Humam AM, Loh TC, Foo HL, Izuddin WI, Awad EA, Idrus Z, et al.

Animals (Basel), 2020 Jun 05;10(6).
PMID: 32516896 DOI: 10.3390/ani10060982

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.
Fulltext Chemical Compositions of Brown and Green Seaweed, and Effects on Nutrient Digestibility in Broiler Chickens

Azizi MN, Loh TC, Foo HL, Akit H, Izuddin WI, Shazali N, et al.

Animals (Basel), 2021 Jul 20;11(7).
PMID: 34359273 DOI: 10.3390/ani11072147

This study aimed to analyse the nutritional properties, apparent ileal digestibility (AID) and apparent metabolisable energy (AME) of broiler chickens fed with brown seaweed (BS) and green seaweed (GS). Proximate analysis was performed to determine the nutrient composition of seaweed. The amino acids were determined using high-performance liquid chromatography (HPLC), and atomic absorption spectroscopy was used to determine the minerals content. The gross energy (GE) was determined using a fully automatic bomb calorimeter, and the AME value was calculated. Titanium dioxide (TiO2) was used as an indigestible marker to calculate the AID. A digestibility trial was conducted to investigate the effects of seaweeds on crude protein (CP), crude fibre (CF), ether extract (EE), dry matter (DM), organic matter (OM), amino acids (AA) and minerals digestibility, and AME on broiler chickens. Thirty-six broiler chickens were randomly distributed into two dietary treatment groups with six replicates and three birds per replicate. Results showed that brown and green seaweed was a source of macro and micronutrients. For the AME and AID of seaweed-based diets, the results showed that the AME value for BS and GS was 2894.13 and 2780.70 kcal/kg, respectively. The AID of BS and GS was 88.82% and 86.8% for EE, 82.03% and 80.6% for OM, 60.69% and 57.80% for CP, 48.56 and 44.02% for CF, and 17.97 and 19.40% for ash contents, respectively. Meanwhile, the AID of CP and CF was significantly higher for BS compared to the GS. Findings showed that the AID of various AA was 40.96 to 77.54%, and the AID of selected minerals (Ca, Na, K, Mg, Zn, Cu, Fe) for both BS and GS groups were above 90%.
Effect of Brachiaria decumbens as a novel supplementation on the production performance of broiler chickens

Alghirani MM, Chung ELT, Kassim NA, Ong YL, Jesse FFA, Sazili AQ, et al.

Trop Anim Health Prod, 2022 Nov 18;54(6):386.
PMID: 36399259 DOI: 10.1007/s11250-022-03384-4

The primary goal of this research was to elucidate the novel influence of Brachiaria decumbens supplementation on broiler chicken growth performance, nutritional digestibility, cecal microbiota, intestinal histomorphology, carcass characteristics, and meat quality. A total of 300 male day-old Ross 308 broiler chickens were randomly subjected to six different treatment groups having five replicates per treatment with 10 birds in each replicate. In treatment 1, broiler chickens were fed commercial diets with no added additives; in treatment 2, broiler chickens were offered commercial diets containing 100 mg/kg of the antibiotic oxytetracycline. However, in treatments 3, 4, 5, and 6, broiler chickens received similar commercial diets supplemented with 25 mg/kg, 50 mg/kg, 75 mg/kg, and 100 mg/kg of B. decumbens ground leaf powder, respectively, without antibiotics. Throughout the 42-day trial, the body weight gain and total feed intake for each replicate were recorded every week to determine the growth performance. Then, on 21th and 42nd day, ten broilers from each treatment (two in each replicate) were randomly selected and slaughtered to assess the digestibility of nutrients, histomorphology of the small intestine, the population of the cecal microbiota, carcass characteristics, as well as quality of both breast and drumstick muscle. There were differences (p

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