• Journal of Nutrition and Health
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• v.53 no.4
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• pp.390-405
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• 2020

Purpose: The effect of prebiotics intake after administration of a synbiotics mixture (a probiotic, Bifidobacterium longum, and a prebiotic, xylooligosaccharide containing sugar [XOS]) on human intestinal microflora and defecation characteristics was investigated in a randomized controlled trial. Methods: Twenty-five healthy young volunteers (11 males and 14 females) were randomly assigned to 2 groups (BL2XO2 and BL2XO6). The synbiotics mixture was orally administered to both groups for 2 weeks, and the prebiotics were subsequently administered to the BL2XO6 group for 4 additional weeks. The daily dose of the synbiotics mixture comprised 10¹⁰ colony-forming unit of Bifidobacterium longum and 10 g of XOS, and during the prebiotics period, the daily dose of prebiotics comprised only 10 g of XOS. The fecal pH, microflora, and defecation characteristics were analyzed at baseline and at weeks 1, 2, 4, and 6. Results: The counts of B. longum and Bifidobacterium spp. in the BL2XO6 group exhibited a steady, increasing trend during the synbiotics and prebiotics periods, whereas those of the BL2XO2 group exhibited considerable variation in each week of the study period. Although there was no significant difference, the counts of fecal Bifidobacterium in the BL2XO6 group tended to be higher than those of the BL2XO2 group at week 6. The growth of Lactobacillus spp. exhibited a time-dependent variation, peaking at week 6 in both groups. Low counts of Clostridium spp. were observed after treatment with the synbiotics and prebiotics in the BL2XO6 group (p < 0.05) throughout the study, whereas the inhibitory effect on Clostridium spp. was maintained only during the synbiotics period in the BL2XO2 group. The defecation characteristics did not differ between the two groups. Conclusion: Administration of XOS after a synbiotics mixture containing B. longum and XOS can exert a prebiotic effect in healthy young volunteers by stimulating Bifidobacteriun spp. growth and inhibiting growth of Clostridium spp.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.24-29
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• 2013

In the present study, the effects of prebiotics and prebiotics+probiotics on intestinal microflora and fermentation products were evaluated in a pig in vitro fermentation model. The substrates used in this study were iso-malto oligosaccharide (IMO), partially digested chicory-inulin (CI), raffinose (RA), and cyclodextrin (CD) as prebiotics and Lactobacillus reiteri as probiotics. For a pig in vitro fermentation, the experimental diet for growing pigs was predigested using digestive enzymes secreted by small intestine and this hydrolyzed diet was mixed with a buffer solution containing 5% fresh swine feces. The mixture was then incubated with either prebiotics or prebiotics+probiotics for 24 h. Samples were taken at 24 h, and viable counts of microflora, gas, pH, volatile organic compounds (VOCs) and short-chain fatty acid (SCFA) were analyzed. The viable count of Enterobacteriaceae was significantly decreased (p<0.001) in all treatments containing prebiotics and prebiotics+probiotics when compared to the control. However, the number of lactic acid bacteria increased in the prebiotics and prebiotics+probiotics treatment. The pH values in the fermentation fluid decreased in all treatments when compared to the control, and their effects were greater in the prebiotics+probiotics group than prebiotics group. Fermentation with prebiotics resulted in a reduction in malodorous compounds such as ammonia, hydrogen sulfide and skatole when compared to the prebiotics+probiotics group. Short-chain fatty acid production was also higher for treatment with prebiotics+probiotics than treatment with prebiotics. In conclusion, the results of this study demonstrated that fermentation with prebiotics was effective in reducing the formation of malodorous compounds and prebiotics+probiotics was effective in increasing lactic acid bacteria and SCFA and reducing the pH. Moreover, further studies will be needed to determine whether the results observed in the in vitro model would occur in pigs that ingest these prebiotics or probiotics.

• Korean Journal of Microbiology
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• v.50 no.4
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• pp.296-301
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• 2014

Rotavirus is a major cause of acute gastroenteritis in young children in developed and developing countries. The use of probiotics for the treatment of gastrointestinal diseases is both safe and easily accessible. In this study, we evaluated the anti-rotaviral activities of probiotic mixtures in a Sprague-Dawley rat. 24 litters with their dams were randomly assigned to four groups; placebo, phosphate buffered saline (PBS), and two probiotic mixture (PRO-1 and PRO-2) groups. All rats were inoculated with rotavirus at dose of 8 log plaque forming units per rat at 5 days old. Animals in the PRO-1 and PRO-2 groups were orally administered probiotic mixtures 1 or 2, respectively, at a dose of 8 log colony forming units daily during 4 days. For control purposes, placebo and PBS groups were orally administered the same amount of placebo (containing maltose and polydextrose) or PBS once daily for 4 days, respectively. Antiviral analysis was performed by real-time quantitative PCR (RT-qPCR) and observing intestinal villi. As a result, weights of small intestines were greater in the PRO-1, PRO-2 groups than in control groups. Villi were short and villous epithelial necrosis was exhibited in control groups, but these morphological changes were not observed in PRO-1, PRO-2 treated rats. RT-qPCR analysis showed that VP7 gene level of rotavirus in fecal samples and small intestinal epithelial cells were lower in the PRO-1 and PRO-2 groups. These findings suggest that probiotic mixtures may be useful probiotics for the treatment of or as alternative therapies for rotaviral gastroenteritis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.10
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• pp.1484-1490
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• 2014

In the present study, we examined the effects of a mixture of three strains, Lactobacillus plantarum CECT 7527, 7528, and 7529 (L. plantarum mixture), on body weight and lipid metabolism in Sprague-Dawley rats fed a high-fat diet. Rats were fed a high-fat diet and subjected to oral gavage with vehicle or the L. plantarum mixture ($0.6{\times}10^9$, $1.2{\times}10^9$, $2.4{\times}10^9$ colony-forming units (CFU)/day/rat, respectively) for 8 weeks. In rats fed a high-fat diet, oral administration of $2.4{\times}10^9CFU/day$ of the L. plantarum mixture significantly reduced body weight gain as well as weights of liver and epididymal fat. Leptin levels in sera were significantly reduced by oral administration of $2.4{\times}10^9CFU/day$ of the L. plantarum mixture. The L. plantarum mixture ($1.2{\times}10^9$ or $2.4{\times}10^9CFU/day$) also reduced the concentrations of total cholesterol and LDL-cholesterol in sera when it administered orally. Further, the L. plantarum mixture significantly reduced the atherogenic index and cardiac risk factor. In addition, oral administration of $2.4{\times}10^9CFU/day$ of the L. plantarum mixture markedly reduced levels of total lipids, triglycerides, and total cholesterol in the liver. The results of this study indicate that the L. plantarum mixture may exhibit anti-obesity and cholesterol-lowering effects, which suggest that the L. plantarum mixture has the potential to be a probiotic in the management of obesity and hypercholesterolemia.

• Food Science of Animal Resources
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• v.33 no.2
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• pp.229-238
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• 2013

Lactic acid bacteria (LAB) are added to different food products for a long time due to health beneficial effects on human host. LAB is applied in dairy products, such as yoghurt, cheese, and various fermented products, and also in non-dairy products, such as sausages. However, reaching the human gut alive as well as in a sufficient cell amount to exert positive health effects is still a big challenge, due to LAB sensitive character and vulnerability against harsh and detrimental conditions in human digestive system. Keeping physiological activity of sensitive LAB strains alive is for the formulation of novel food products with a probiotic health claim of utmost interest, thus microencapsulation has been applied and investigated as a promising technique for a good and reliable protection. Microencapsulation allows reduced cell injury or cell loss by retaining cells within the encapsulating membrane and can be enforced by spray-drying, emulsion, extrusion, and a range of other technologies in combination with an appropriate coating material, such as alginate, chitosan, and mixture of these two polymers. In this review, established and well-studied microencapsulation techniques with their favored coating materials, as well as the recent applications of microencapsulated LAB into dairy products will be discussed.

• Journal of Microbiology and Biotechnology
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• v.13 no.2
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• pp.276-281
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• 2003

The adhesion of probiotic bacteria to the intestinal mucosa is one of the desirable properties for their colonization in the intestinal tract, where these bacteria constantly compete with other bacteria. The adhesion of different strains of bifidobacteria to Caco-2 cells was compared. Among the strains examined, BGN-4 showed the highest adhesion level and the greatest cell surface hydrophobicity (CSH). No close relationship was found between the adhesion and CSH of the strains. Upon protease and heat treatment, the adhesion of the BGN-4 to the Caco-2 cells decreased significantly. The cells grown at $42^{\circ}C$ showed a lower CSH and self-aggregation levels than cells grown at $37^{\circ}C$. The treatment of EGTA did not have any effect on the adhesion. The degree of adhesion did not differ among the experimental groups in which galactose, mannose, or fucose were added in the adhesion assay mixture. The results suggest that the adhesion of the Bifidobacterium to the epithelial cells may be affected by the composition and structure of the cell membrane and interacting surfaces.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.5
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• pp.655-661
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• 2011

Two experiments were conducted to investigate the effect of dietary supplementation of Bacillus, Saccharomyces and lactic acid bacteria (LAB) on performance and nutrient digestibility in grower and finisher pigs. In Exp. 1, 80 pigs (32 females and 48 males), $28.7{\pm}0.9\;kg$ body weight (BW), were randomly divided into 4 treatment groups balanced for sex and weight (5 pigs per pen, 4 pens per treatment). They were fed one of four diets: a basal grower (20-50 kg BW) and finisher (>50 kg BW) diet without any addition of probiotic or antibiotic (diet C), the basal diet supplemented with Bacillus subtilis H4 (diet B), diet B supplemented with Saccharomyces boulardi Sb (diet BS) and diet BS supplemented with a LAB complex (diet BSL). The LAB complex consisted of Enterococcus faecium 6H2, Lactobacillus acidophilus C3, Pediococcus pentosaceus D7, and Lactobacillus fermentum NC1. In Exp. 2, 16 male pigs, $29.2{\pm}0.8\;kg$ BW, were kept in individual pens and divided into 4 groups (4 pigs in each group). All 4 groups were given exactly the same growing-period diets (diet C, B, BS and BSL) as in Exp 1. The total faeces and urine were collected during 5 days (day 20-24) to determine nitrogen retention and total tract digestibility. In the growing period, average daily feed intake (ADFI), average daily gain (ADG) and feed conversion ratio (FCR) were not affected by diet B and BS (p>0.05), but ADG increased (+5.9%) (p<0.05) and FCR improved (+5.9%) (p<0.05) on diet BSL compared with the control, although ADFI was not different (p>0.05). Digestibility of crude protein and organic matter was higher (p<0.05) in diet BSL and digestibility of crude fibre was higher (p<0.05) in diet BS and BSL than in diet C. Nitrogen retention was not affected by diet (p>0.05). The faecal LAB counts were increased in grower pigs fed diet BSL (p<0.05) and faecal E. coli counts were decreased in pigs fed diets BS and BSL (p<0.05). In the finishing period, no effects of diet were found in ADFI, ADG, FCR, nutrient digestibility, and nitrogen retention (p>0.05). Faecal LAB and E. coli counts in the finisher pigs were not affected by diet (p>0.05). In conclusion, the current study demonstrates that a mixture of bacteria and yeast has the potential to be used as a probiotic dietary supplement in grower pigs.

• Korean Journal of Microbiology
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• v.47 no.3
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• pp.231-240
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• 2011

The objective of this study was to evaluate the acid and bile tolerance, bile salt hydrolase (BSH) activity, and cholesterol assimilation ability of lactic acid bacteria isolated from mustard leaf kimchi. MLK11, MLK22, MLK27, MLK41, and MLK67 were relatively acid- and bile-tolerant strains, with more than $10^5$ CFU/ml after incubation in simulated gastric juice and intestinal fluid, while MLK53 was the most sensitive strain to acid and bile. Strains MLK22 and MLK67 deconjugated the highest level of sodium glycocholate with more than 3.5 mM of cholic acid released, while deconjugation was lowest by strains MLK13 and MLK41 which released only 1.35 mM and 1.16 mM, respectively. Specially, strains MLK22 and MLK67 showed higher deconjugation of sodium glycocholate compared to sodium taurocholate and conjugated bile mixture. Although strains MLK22 and MLK67 exhibited maximal BSH activity at the stationary phase, MLK22 had somewhat higher total BSH activity compared to MLK67 towards both sodium glycocholate and sodium taurocholate. Meanwhile, cholesterol removal varied among tested strains (p<0.05) and ranged from 5.22 to 39.16 ${\mu}g$/ml. Especially, MLK67 strain assimilated the highest level of cholesterol in media supplemented with 0.3% oxgall, cholic acid, and taurocholic acid (p<0.05). According to physiological and biological characteristics, pattern of carbohydrate fermentation, and 16S rDNA sequence, strain MLK67 that may be considered as probiotic strain due to acid and bile tolerance and cholesterol-lowering effects was identified as Pediococcus pentosaceus MLK67.

• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.718-731
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• 2018

The beneficial effects of lactic acid bacteria (LAB) have been intensively investigated in recent decades with special focus on modulation of the host intestinal microbiota. Numerous discoveries of effective probiotics are driven by a significantly increasing demand for dietary supplements. Consequently, technological advances in the large-scale production and lyophilization are needed by probiotic-related industries for producing probiotic LAB for commercial use. Our study had a dual objective, to determine the optimum growth medium composition and to investigate appropriate cryoprotective additives (CPAs) for Lactobacillus salivarius, and compare its responses with other Lactobacillus species. The one-factor-at-a-time method and central composite design were applied to determine the optimal medium composition for L. salivarius cultivation. The following composition of the medium was established (per liter): 21.64 g maltose, 85 g yeast extract, 1.21 ml Tween 80, 6 g sodium acetate, $0.2g\;MgSO_4{\cdot}7H_2O$, $0.02g\;MnSO_4{\cdot}H_2O$, $1g\;K_2HPO_4$, $1.5g\;KH_2PO_4$, $0.01g\;FeSO_4{\cdot}7H_2O$, and 1 g sodium citrate. A cryoprotective additive combination comprising 10% (w/v) skim milk and 10% (w/v) sucrose supplemented with 2.5% (w/v) sodium glutamate was selected for L. salivarius, and its effectiveness was confirmed using culture-independent methods in the freeze-dried cells of the Lactobacillus strains. In conclusion, the optimized medium enhanced the species-specific cultivation of L. salivarius. On the other hand, the cryoprotective effects of the selected CPA mixture may also be dependent on the bacterial strain. This study highlights the necessity for precise and advanced processing techniques for large-scale production of probiotics in the food and feed industries.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.4
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• pp.493-499
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• 2011

The objective of this study was to find the way to prolong the storage time of sawdust-based oyster mushroom (Pleurotus osteratus) spent substrate (OMSS) by fermenting with potential probiotic microorganisms to recycle the otherwise waste of mushroom farms. To this purpose, lactic acid bacteria (LAB) were screened to select the best lactic acid-producing strains. Three strains of LAB (Lactobacillus plantarum Lp1', Pediococcus acidilacticii Pa193, L. plantarum Lp2M) were selected and in mixture they lowered the pH of the fermented OMSS to 3.81. fOMSS (fermented sawdust-based oyster mushroom spent substrate) could be stored at room temperature for at least 17 days without any deterioration of feed quality based on the pH, smell, and color. In dry matter disappearance rate in situ, commercial TMR (total mixed ration), OMSS and OMMM (oyster mushroom mycelium mass) showed no significant differences between the samples after 6, 12 and 24 h incubation except for 48 h. Two separate field studies were performed to test the effects of fOMSS supplement on the growth performance of postweaning Holstein calves. Field trials included groups of animals feeding calf starter supplemented with: Control (no supplement), AB (colistin 0.08% and oxyneo 110/110 0.1%), fOMSS (10% fOMSS) and fConc (10% fermented concentrate) and DFM (direct-fed microbials, average $10^9$ cfu for each of three LAB/d/head). Growth performance (average daily gain and feed efficiency) of the fOMSS supplement group was higher than that of AB followed by fConc and DFM even though there was no statistically significant difference. The Control group was lower than any other group. Various hematological values including IgG, IgA, RBC (red blood cell), hemoglobin, and hematocrit were measured every 10 days to check any unusual abnormality for all groups in trial I and II, and they were within a normal and safe range. Our results suggest that sawdust-based OMSS could be recycled after fermentation with three probiotic LAB strains as a feed supplement for post-weaning calves, and fOMSS has the beneficial effects of an alternative to antibiotics for a growth enhancer in dairy calves.