A study was conducted to determine the effects of adherent Lactobacillus culture on growth performance, intestinal microbial population, and serum cholesterol level of broilers. Four dietary treatments, consisting of the basal diet (control), basal diet + 0.05, 0.10, or 0.15% Lactobacillus culture (LC), were fed to 2,000 Arbor Acres broiler chicks from 1 to 42 d of age (DOA). The chicks were randomly assigned to 40 cages (50 chicks per cage, 10 cages per diet). The experimental period was 42 d. Body weights and feed to gain ratio were measured at 21 and 42 DOA. The intestinal microbial populations and serum cholesterol levels were determined at 10, 20, 30, and 40 DOA. The results showed that body weights and feed to gain ratios were improved significantly (P < 0.05) when compared to control broilers for broilers fed diets containing 0.05 or 0.10% LC, but not 0.15% LC, at 21 and 42 DOA. Coliform counts in the cecum of birds receiving 0.05% LC at 10, 20, and 30 DOA, and 0.10% at 10 and 20 DOA were significantly lower (P < 0.05) than those of the control birds. The total aerobes, total anaerobes, lactobacilli, and streptococci in the small intestines and ceca of the control birds were not significantly different from those of the treated groups. Serum cholesterol levels were significantly lower (P < 0.05) in broilers fed the three diets containing LC at 30 DOA, and in the birds fed 0.05 or 0.10% LC at 20 DOA.
Probiotic organism Bifidobacteria was isolated from the faeces of breast-fed infants at Universiti Putra Malaysia. Trypticase phytone peptone yeast extract agar (TPY) was used as a selective media for the isolation. Morphological examination of the isolates indicated that Bifidobacteria was Gram-positive rods in nature, curved with characteristics of V and Y shapes. The organisms were non-catalase producing, non-nitrate reducing, non-motile, had an absence of indole and were unable to liquify gelatin. The ratios of acetic and lactic acids were determined using high performance liquid chromatography (HPLC). Using carbohydrate fermentation profile test API-CH-50 kits, 20 Bifidobacteria strains had been identified: they were the species of Bifidobacteria infantis and two different sub-species, mainly infantis and lacentis. Based on a wide zone of inhibition, three suitable strains of B. infantis, Bifi-11, Bifi-19 and Bifi-20, were tested in weaning foods for antimicrobial activity towards two human pathogens: E. coli-0157 (World Health Organization) and Salmonella typhimurium S-285. The pH, titratable acidity of weaning foods and total colony count for Bifidobacteria, enteropathogenic Escherichia coli and S. typhimurium were recorded at 3-h intervals for 30 h. It was found that after 9 h of incubation of weaning foods, the pH declined to < 3.6 from pH 6.0, whereas titratable acidity increased from 0.026 to 0.08%. It was indicated that Bifidobacteria inhibited E. coli better than did S typhimurium due to low pH. After 24 h of incubation, approximately 98% of E. coli was inhibited by Bifidobacteria. It is suggested that the inhibitory effect of Bifidobacteria strains in weaning foods towards the growth of enteropathogenic E. coli and S. typhimurium was solely due to low pH and the production of volatile acid components by the organism.
1. Hubbard x Hubbard (HH) and Shaver x Shaver (SS) chicks given a dietary supplement of either 50 mg/kg oxytetracycline (OTC) or 1 g/kg Lactobacillus culture (LC) were exposed to 36 +/- 1 degrees C for 3 h daily from day (d) 21 to 42. 2. Prior to heat treatment, body weight (d 21) and weight gain (d 1 to d 21) of OTC and LC birds were greater than those fed the control diet. Chicks given LC had the best food efficiency followed by OTC and control birds during d 1 to d 21. Body weight (d 1 and d 21) and weight gain (d 1 to d 21) were greater for HH tlhan SS chicks. 3. After 3 weeks of heat exposure, birds receiving the LC diet had greater body weight and weight gain, higher food intake and lower food efficiency than OTC and control chicks. 4. Antibody production against Newcastle discase vaccine on d 21 was not affected by strain or diet. On d 42, while diet had negligible effect on this variable among the SS broilers, HH birds fed LC had higher antibody production than those on the control diet. 5. Neither strain nor diet had a significant effect on mortality.
We evaluated the efficacy of supplementation of active Mitsuokella jalaludinii culture (AMJC) on the growth performance, nutrient use, and mineral concentrations in tibia bone and plasma of broiler chickens fed corn-soybean meal diets. Dietary treatments included low-nonphytate P (NPP) feed (containing 0.24% and 0.232% NPP for chicks from 1 to 21 and 22 to 42 d of age, respectively), low-NPP feed added with different levels of AMJC (equivalent to 250, 500, 750, and 1,000 U phytase/kg of feed), and normal-NPP feed (containing 0.46 and 0.354% NPP for chicks from 1 to 21 and 22 to 42 d of age, respectively). Supplementation of AMJC to low-NPP feed increased (P < 0.05) weight gain and feed intake and decreased (P < 0.05) feed:gain ratio of chickens during the whole experiment (Days 1 to 42). Supplementation of AMJC increased (P < 0.05) the AME value, digestibility of DM and CP, and retention of P, Ca, and Cu. Mn retention in broilers was only increased (P < 0.05) by AMJC supplementation from 18 to 20 d of age, and Zn retention was improved (P < 0.05) only at a high level of AMJC (equivalent to 1,000 U phytase/kg of feed) supplementation. Chicks fed low-NPP feed added with AMJC had similar tibia ash percentages as those fed the normal-NPP diet. Generally, supplementing AMJC to low-NPP feed increased (P < 0.05) Ca, decreased significantly (P < 0.05) Mn and Cu, but did not affect Zn and P concentrations in tibia ash. Supplementing AMJC also increased (P < 0.05) plasma P but had no effect on plasma Ca or Mn. Plasma Zn concentration was increased only when a high level of AMJC (equivalent to 1,000 U phytase/kg of feed) was used. In conclusion, AMJC supplementation to low-NPP feed improved growth performance; AME value; digestibility of CP and DM; use of Ca, P, and Cu; and bone mineralization.
This study was undertaken to optimize yeast extract, glucose, and vitamin concentrations; and also culture pH for maximizing the growth of a probiotic bacterium, Lactobacillus rhamnosus, and to assess the effects of these factors by using response surface methodology. A central composite design was used as an experimental design for the allocation of treatment combinations. A polynomial regression model with cubic and quartic terms was used for analysis of the experimental data. It was found that the effects involving yeast extract, glucose, vitamins and pH on the growth of L. rhamnosus were significant, and the strongest effect was given by the yeast extract concentration. Estimated optimum conditions of the factors for the growth of L. rhamnosus are as follows: pH=6.9; vitamin solution=1.28% (v/v); glucose=5.01% (w/v) and yeast extract=6.0% (w/v).
This study evaluated the effect of a Probiotic mouthrinse containing nisin, a bacteriocin extracted from Lactococcus lactis on dental plaque and gingivitis in young adult population. A group of 32 subjects were randomly assigned into two groups of 16 each. The first group started using the control mouthrinse (placebo) for 2 weeks followed by a washout period of 4 weeks. This group then used the test mouthrinse (Probiotic) for a further duration of 2 weeks. The second group followed a similar protocol as the first except that this group started with the test mouthrinse (Probiotic). Plaque Index (PI) and Gingival Index (GI) were recorded at baseline and after 2 weeks for each group. All subjects were given full mouth prophylaxis after each measurements. The results of this study showed that rinsing with Probiotic mouthrinse resulted in a statistically significant reduction of plaque accumulation and gingivitis compared to rinsing with placebo. The results indicated that Probiotic mouthrinse containing nisin had the potential of inhibiting plaque accumulation and was effective in reducing gingivitis.
To assess the safety of Bifidobacterium pseudocatenulatum G4 in BALB/c mice that involves examination of bacterial translocation, changes in the internal organs and histology of the intestinal lining.
The copy numbers of 16S rRNA genes in 12 probiotic Lactobacillus strains of poultry origin were analyzed. Genomic DNA of the strains was digested with restriction endonucleases that do not cut within the 16S rRNA gene of the strains. This was followed by Southern hybridization with a biotinylated probe complementary to the 16S rRNA gene. The copy number of the 16S rRNA gene within a Lactobacillus species was found to be conserved. From the hybridization results, Lactobacillus salivarius I 24 was estimated to have seven copies of the 16S rRNA gene, Lactobacillus panis C 17 to have five copies and Lactobacillus gallinarum strains I 16 and I 26 four copies. The 16S rRNA gene copy numbers of L. gallinarum and L. panis reported in the present study are the first record. Lactobacillus brevis strains I 12, I 23, I 25, I 211, I 218 and Lactobacillus reuteri strains C 1, C 10, C 16 were estimated to have at least four copies of the 16S rRNA gene. In addition, distinct rRNA restriction patterns which could discriminate the strains of L. reuteri and L. gallinarum were also detected. Information on 16S rRNA gene copy number is important for physiological, evolutionary and population studies of the bacteria.
Probiotics are live organisms that are primarily used to improve gastrointestinal disorders such as diarrhea, irritable bowel syndrome, constipation, lactose intolerance, and to inhibit the excessive proliferation of pathogenic intestinal bacteria. However, recent studies have suggested that probiotics could have beneficial effects beyond gastrointestinal health, as they were found to improve certain metabolic disorders such as hypertension. Hypertension is caused by various factors and the predominant causes include an increase in cholesterol levels, incidence of diabetes, inconsistent modulation of renin and imbalanced sexual hormones. This review discusses the antihypertensive roles of probiotics via the improvement and/or treatment of lipid profiles, modulation of insulin resistance and sensitivity, the modulation of renin levels and also the conversion of bioactive phytoestrogens as an alternative replacement of sexual hormones such as estrogen and progesterone.
The objective of this study was to evaluate agricultural wastes as immobilizers for probiotics in liquid foods, such as soy milk. Probiotic strains were initially evaluated for acid and bile tolerance and the ability to produce alpha-galactosidase. Rinds of durian, mangosteen, and jackfruit were dried, ground, and sterilized prior to immobilization of selected strains ( Lactobacillus acidophilus FTDC 1331, L. acidophilus FTDC 2631, L. acidophilus FTDC 2333, L. acidophilus FTDC 1733, and Lactobacillus bulgaricus FTCC 0411). Immobilized cells were inoculated into soy milk, and growth properties were evaluated over 168 h at 37 degrees C. Soy milk containing free cells without agrowastes was used as the control. Immobilized probiotics showed increased growth, greater reduction of stachyose, sucrose, and glucose, higher production of lactic and acetic acids, and lower pH in soy milk compared to the control. The results illustrated that agrowastes could be used for the immobilization of probiotics with enhanced growth, utilization of substrates, and production of organic acids.
Different probiotic strains are incorporated into cultured-milk drinks by respective manufacturers with the common aim of providing health benefits to the consumers. Four common cultured-milk products (brands N, S, V and Y) were evaluated for their quality of probiotic strains used, based on the susceptibility of the probiotics to various pH levels simulated to mimic the gastrointestinal system. Results showed that brands Y and V have higher initial probiotic inoculum compared to brands N and S although probiotics from brands N and S were more tolerant to pH 3. Generally, all probiotic strains preferred higher pH with highest viability of Lactobacillus spp., Bifidobacterium spp., and Streptococcus thermophilus observed at pH 8.1. Our study also discovered that among the four brands tested, brand V contained probiotic strains which are most likely to remain viable after passage through the gastrointestinal system.
Bile salt deconjugation by Lactobacillus strains is often closely linked to bile tolerance and survival of the strains in the gut and lowering of cholesterol in the host. The present study investigated the deconjugation of bile salts and removal of cholesterol by 12 Lactobacillus strains in vitro. The 12 strains were previously isolated from the gastrointestinal tract of chickens.
Soy products have attracted much attention lately as carriers for probiotics. This study was aimed at enhancing the growth of probiotics in soymilk via supplementation with prebiotics.
Ten strains of Lactobacillus were evaluated for their viability in soymilk. Lactobacillus acidophilus ATCC 314, L. acidophilus FTDC 8833, L. acidophilus FTDC 8633 and L. gasseri FTDC 8131 displayed higher viability in soymilk and were thus selected to be evaluated for viability and growth characteristics in soymilk supplemented with B-vitamins. Pour plate analyses showed that the supplementation of all B-vitamins studied promoted the growth of lactobacilli to a viable count exceeding 7 log CFU/ml. alpha-Galactosidase specific activity of lactobacilli as determined spectrophotometrically showed an increase upon supplementation of B-vitamins. High-performance liquid chromatography analyses revealed that this led to increased hydrolysis of soy oligosaccharides and subsequently higher utilization of simple sugars. Production of organic acids as determined via high-performance liquid chromatography also showed an increase, accompanied by a decrease in pH of soymilk. Additionally, the supplementation of B-vitamins also promoted the synthesis of riboflavin and folic acid by lactobacilli in soymilk. Our results indicated that B-vitamin-supplemented soymilk is a good proliferation medium for strains of lactobacilli.
1. An experiment was conducted to determine the effects of supposedly unpleasant physical treatment on broiler performance, small intestinal development and ameliorating role of probiotics. 2. The following treatments were applied from day one: (1) chicks exposed to normal human contact fed basal diet (control); (2) chicks were exposed to unpleasant physical treatment and fed basal diet (UPT-BD); and (3) chicks were exposed to unpleasant physical treatment and fed basal diet supplemented with Lactobacillus (UPT-BDL). Chicks were exposed to UPT from days 1 to 21. Different segments of gastrointestinal tract were sampled at 14, 28, 35 and 42 d of age. 3. Broilers of UPT-BD had lower feed consumption compared with control group at 7 d of age. Overall, UPT-BDL birds showed higher body weight gain (BWG) and better feed conversion ratio (FCR) over the course of the experiment. 4. Birds of UPT-BD had lower concentrations of lactic, propionic and butyric acids in the caecum as compared with other groups at 14 d of age. Acetic acid concentration was profoundly decreased in both UPT groups compared to the control. 5. Duodenal villus height of UPT-BD broilers showed a slight reduction compared to the control and UPT-BDL birds at 14 d of age. Afterwards until day 42, UPT-BDL birds showed the highest villus height among treatments in different parts of the small intestine. 6. The results suggested that, even though UPT did not have significant inhibitory effects on the development of the small intestine and broiler performance, it negatively affected bacterial metabolic end products in the caecum, which could be ameliorated by the addition of Lactobacillus.
The in vitro fermentability of sago (Metroxylon sagu) resistant starch type III (RS(3)) by selected probiotic bacteria was investigated. Sago RS(3) with 12% RS content was prepared by enzymatic debranching of native sago starch with pullulanase enzyme, followed by autoclaving, cooling, and annealing. The fermentation of sago RS(3) by L. acidophilus FTCC 0291, L. bulgaricus FTCC 0411, L. casei FTCC 0442, and B. bifidum BB12 was investigated by observing the bacterial growth, carbohydrate consumption profiles, pH changes, and total short chain fatty acids (SCFA) produced in the fermentation media. Comparisons were made with commercial fructo-oligosaccharide (FOS), Hi-maize 1043, and Hi-maize 240. Submerged fermentations were conducted in 30 mL glass vials for 24 h at 37 degrees C in an oven without shaking. The results indicated that fermentation of sago RS(3) significantly (P < 0.05) yielded the highest count of Lactobacillus sp. accompanied by the largest reduction in pH of the medium. Sago RS(3) was significantly the most consumed substrate compared to FOS and Hi-maizes.
Lactobacillus sp. FTDC 2113, L. acidophilus FTDC 8033, L. acidophilus ATCC 4356, L. casei ATCC 393, Bifidobacterium FTDC 8943 and B. longum FTDC 8643 were incorporated into soymilk supplemented with fructooligosaccharides (FOS), inulin, mannitol, maltodextrin and pectin. The objective of the present study was to evaluate the effects of prebiotics on the bioactivity of probiotic-fermented soymilk. Proteolytic activity was increased in the presence of FOS, while the supplementation of inulin and pectin increased the angiotensin I-converting enzyme inhibitory activity accompanied by lower IC(50) values. The beta-glucosidase activity was also enhanced in the presence of pectin. This led to higher bioconversion of glucosides to aglycones by probiotics, especially genistin and malonyl genistin to genistein. Results from this study indicated that the supplementation of prebiotics enhanced the in-vitro antihypertensive effect and production of bioactive aglycones in probiotic-fermented soymilk. Therefore, this soymilk could potentially be used as a dietary therapy to reduce the risks of hypertension and hormone-dependent diseases such as breast cancer, prostate cancer and osteoporosis.
Among the bacterial fermentation end products in the chicken cecum, butyrate is of particular importance because of its nutritional properties for the epithelial cell and pathogen inhibitory effects in the gut. An in vitro experiment, operated with batch bioreactor, was conducted to quantify butyric-producing bacteria in a simulated broiler cecum supplemented with Lactobacillus salivarius ssp. salicinius JCM 1230 and Lactobacillus agilis JCM 1048 during 24 h of incubation. Selected bacterial species were determined by real-time PCR and short-chain fatty acids and lactate concentrations were monitored. The results showed that after 24 h of incubation, Lactobacillus supplementation significantly increased the number of lactobacilli, bifidobacteria and Faecalibacterium prausnitzii in medium containing cecal content and lactobacilli supplementation (Cc + L) compared with the control (Cc). Addition of lactobacilli did not alter Escherichia coli and Clostridium butyricum, whereas it significantly (P < 0.05) reduced Salmonella in treatment Cc + L compared with the Cc treatment. Propionate and butyrate formation were significantly (P < 0.05) increased in treatment Cc + L as compared with the Cc treatment. Lactate was only detected in treatment containing 2 Lactobacillus strains. After 24 h of incubation, acetate concentration significantly (P < 0.05) decreased in all treatments. It was suggested that lactate produced by Lactobacillus in the cecal content improved the growth of butyric producers such as F. prausnitzii, which significantly increased butyrate accumulation. Additionally, the results showed that butyrate and propionate inhibited Salmonella without influencing the E. coli profile.
The aim of this work was to develop a standard quantitative method to measure the acid tolerance of probiotic cells when exposed to a simulated gastric fluid. Three model strains of different cell concentrations were exposed to a standard simulated gastric fluid of fixed volume. The fluid pH ranged from pH 1.5 to 2.5. In general, the death kinetics followed an exponential trend. The overall death constant, k (d), for all strains was found to be in a power relationship with the pH value and the initial cell concentration, and it can be expressed as k(d)=k(AII) (pH(-9.0)N(0)(-0.19)) where k (AII) is defined as the acid intolerance indicator and N (0) is the initial cell concentration (CFU/ml). This equation was validated with the experimental data with an average R (2) of 0.98. The acid intolerance of cells can be quantitatively expressed by the k (AII) values, where higher value indicates higher intolerance. In conclusion, a standard quantitative method has been developed to measure the acid tolerance of probiotic cells. This could facilitate the selection of probiotic strains and processing technologies.
Cell immobilization is an alternative to microencapsulation for the maintenance of cells in a liquid medium. The objective of this study was to evaluate the effects of agrowastes from durian (Durio zibethinus), cempedak (Artocarpus champeden), and mangosteen (Garcinia mangostana) as immobilizers for lactobacilli grown in soymilk. Rinds from the agrowastes were separated from the skin, dried, and ground (150 microm) to form powders and used as immobilizers. Scanning electron microscopy revealed that lactobacilli cells were attached and bound to the surface of the immobilizers. Immobilized cells of Lactobacillus acidophilus FTDC 1331, L. acidophilus FTDC 2631, L. acidophilus FTDC 2333, L. acidophilus FTDC 1733, and L. bulgaricus FTCC 0411 were inoculated into soymilk, stored at room temperature (25 degrees C) and growth properties were evaluated over 168 h. Soymilk inoculated with nonimmobilized cells was used as the control. Utilization of substrates, concentrations of lactic and acetic acids, and changes in pH were evaluated in soymilk over 186 h. Immobilized lactobacilli showed significantly better growth (P < 0.05) compared to the control, accompanied by higher production of lactic and acetic acids in soymilk. Soymilk containing immobilized cells showed greater reduction of soy sugars such as stachyose, raffinose, sucrose, fructose, and glucose compared to the control (P < 0.05).