• Journal of Animal Science and Technology
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• v.64 no.6
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• pp.1144-1172
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• 2022

Salmonella enterica serovar Typhimurium isolate HJL777 is a virulent bacterial strain in pigs. The high rate of salmonella infection are at high risk of non-typhoidal salmonella gastroenteritis development. Salmonellosis is most common in young pigs. We investigated changes in gut microbiota and biological function in piglets infected with salmonella via analysis of rectal fecal metagenome and intestinal transcriptome using 16S rRNA and RNA sequencing. We identified a decrease in Bacteroides and increase in harmful bacteria such as Spirochaetes and Proteobacteria by microbial community analysis. We predicted that reduction of Bacteroides by salmonella infection causes proliferation of salmonella and harmful bacteria that can cause an intestinal inflammatory response. Functional profiling of microbial communities in piglets with salmonella infection showed increasing lipid metabolism associated with proliferation of harmful bacteria and inflammatory responses. Transcriptome analysis identified 31 differentially expressed genes. Using gene ontology and Innate Immune Database analysis, we identified that BGN, DCN, ZFPM2 and BPI genes were involved in extracellular and immune mechanisms, specifically salmonella adhesion to host cells and inflammatory responses during infection. We confirmed alterations in gut microbiota and biological function during salmonella infection in piglets. Our findings will help prevent disease and improve productivity in the swine industry.

• Journal of Microbiology and Biotechnology
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• v.27 no.11
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• pp.1971-1982
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• 2017

Piliated Lactobacillus rhamnosus (pLR) strains possess higher adherent capacity than non-piliated strains. The objective of this study was to isolate and characterize probiotic pLR strains in human fecal samples. To this end, mouse polyclonal antiserum (anti-SpaA) against the recombinant pilus protein (SpaA) of L. rhamnosus strain GG (LGG) was prepared and tested for its reactivity and specificity. With the anti-SpaA, a method combining the de Man, Rogosa, and Sharpe (MRS) agar plating separation and colony immunoblotting (CIB) was developed to isolate pLR from 124 human fecal samples. The genetic and phenotypic characteristics of the resultant pLR isolates were compared by randomly amplified polymorphic DNA (RAPD) fingerprinting, and examination of adhesion to Caco-2 cells, hydrophobicity, autoaggregation, and in vitro gastrointestinal tolerance. Anti-SpaA specifically reacted with three pLR strains of 25 test strains, as assessed by western blotting, immunofluorescence flow cytometry, and immunoelectron microscopy (IEM) assays. The optimized MRS agar separation plus anti-SpaA-based CIB procedure could quantitatively detect $2.5{\times}10^3CFU/ml$ of pLR colonies spiked in $10^6CFU/ml$ of background bacteria. Eight pLR strains were identified in 124 human fecal samples, and were confirmed by 16S RNA gene sequencing and IEM identification. RAPD fingerprinting of the pLR strains revealed seven different patterns, of which only two isolates from infants showed the same RAPD profiles with LGG. Strain PLR06 was obtained with high adhesion and autoaggregation activities, hydrophobicity, and gastrointestinal tolerance. Anti-SpaA-based CIB is a rapid and inexpensive method for the preliminary screening of novel adherent L. rhamnosus strains for commercial purposes.

• Journal of Ginseng Research
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• v.39 no.2
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• pp.183-187
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• 2015

Background: Gut microflora play a crucial role in the biotransformation of ginsenosides to compound K (CK), which may affect the pharmacological effects of ginseng. Prebiotics, such as NUTRIOSE, could enhance the formation and consequent absorption of CK through the modulation of gut microbial metabolic activities. In this study, the effect of a prebiotic fiber (NUTRIOSE) on the pharmacokinetics of ginsenoside CK, a bioactive metabolite of ginsenosides, and its mechanism of action were investigated. Methods: Male Sprague-Dawley rats were given control or NUTRIOSE-containing diets (control diet + NUTRIOSE) for 2 wk, and ginseng extract or vehicle was then orally administered. Blood samples were collected to investigate the pharmacokinetics of CK using liquid chromatography-tandem mass spectrometry. Fecal activities that metabolize ginsenoside Rb1 to CK were assayed with fecal specimens or bacteria cultures. Results: When ginseng extract was orally administered to rats fed with 2.5%, 5%, or 10% NUTRIOSE containing diets, the maximum plasma concentration ($C_{max}$) and area under the plasma concentration-time curve values of CK significantly increased in a NUTRIOSE content-dependent manner. NUTRIOSE intake increased glycosidase activity and CK formation in rat intestinal contents. The CK-forming activities of intestinal microbiota cultured in vitro were significantly induced by NUTRIOSE. Conclusion: These results show that prebiotic diets, such as NUTRIOSE, may promote the metabolic conversion of ginsenosides to CK and the subsequent absorption of CK in the gastrointestinal tract and may potentiate the pharmacological effects of ginseng.

• Journal of Animal Science and Technology
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• v.63 no.6
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• pp.1314-1327
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• 2021

Bacillus is characterized by the formation of spores in harsh environments, which makes it suitable for use as a probiotic for feed because of thermostability and high survival rate, even under long-term storage. This study was conducted to investigate the effects of Bacillus-based probiotics on growth performance, nutrient digestibility, intestinal morphology, immune response, and intestinal microbiota of weaned pigs. A total of 40 weaned pigs (7.01 ± 0.86 kg body weight [BW]; 28 d old) were randomly assigned to two treatments (4 pigs/pen; 5 replicates/treatment) in a randomized complete block design (block = BW and sex). The dietary treatment was either a typical nursery diet based on corn and soybean meal (CON) or CON supplemented with 0.01% probiotics containing a mixture of Bacillus subtilis and Bacillus licheniformis (PRO). Fecal samples were collected daily by rectal palpation for the last 3 days after a 4-day adaptation. Blood, ileal digesta, and intestinal tissue samples were collected from one pig in each pen at the respective time points. The PRO group did not affect the feed efficiency, but the average daily gain was significantly improved (p < 0.05). The PRO group showed a trend of improved crude protein digestibility (p < 0.10). The serum transforming growth factor-β1 level tended to be higher (p < 0.10) in the PRO group on days 7 and 14. There was no difference in phylum level of the intestinal microbiota, but there were differences in genus composition and proportions. However, β-diversity analysis showed no statistical differences between the CON and the PRO groups. Taken together, Bacillus-based probiotics had beneficial effects on the growth performance, immune system, and intestinal microbiota of weaned pigs, suggesting that Bacillus can be utilized as a functional probiotic for weaned pigs.

• Korean Journal of Poultry Science
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• v.49 no.2
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• pp.125-137
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• 2022

This study aimed to determine the effects of dietary microalgae (Tetradesmus sp. (TO)) on intestinal immunity and microbiota of pre-starter broilers. One hundred and twenty 1-day-old birds (Ross 308) were allocated to two dietary treatment groups with six blocks in a randomized complete block design. The two experimental diets consisted of a corn-soybean meal-based basal diet and a diet with 0.5% TO powder instead of cornstarch in the basal diet. After feeding the experimental diets for ten days, all birds' body weight and feed intake were measured, and representative eight birds were selected from each treatment group. Small intestinal lamina propria cells were isolated using flow cytometry to examine the frequency of immune cells. Cecal feces were harvested for 16s rRNA gut microbiota analysis and fecal IgA levels. Here, we found that 0.5% TO supplementation increased CD3⁺CD4⁺ T cells in the small intestine, but decreased CD3⁺CD8⁺ T cells in the small intestine. Gut microbial analysis showed that TO supplementation significantly increased the alpha diversity of the gut microbiome. Taxonomic analysis showed that TO treatment increased the abundance of Firmicutes and decreased that of Bacteroidetes at the phylum level. The distribution of Enterobacteriaceae containing many harmful bacteria at the family level, was lower in the TO group. In the LEfSe analysis, the TO group had a significantly enriched abundance of Agathobaculum at the genus level. Overall, results show that Tetradesmus sp. supplementation influences intestinal T-cell immunity and induces the expansion of beneficial gut microbes in pre-starter broiler chickens.

• Journal of Species Research
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• v.9 no.1
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• pp.35-45
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• 2020

In 2016 and 2017, as part of a comprehensive investigation to identify the prokaryotic species in Korea, a total of 12 bacterial strains were isolated from the gastrointestinal tract and/or fecal samples of four endangered species, including reptile, bird, and marine and terrestrial mammals. Phylogenetic analysis with the 16S rRNA gene sequence was used to assign these strains to the phyla, Firmicutes, Actinobacteria or Proteobacteria. Furthermore, most of the strains Firmicutes belonged to the order Lactobacillales. Interestingly, 12 of the isolated strains have not been previously reported from the Korean Peninsula. Also, based on their high 16S rRNA gene sequence similarities(>98.7%) and formation of strong monophyletic clades with the closest type species, each isolated strain of isolates was assigned to an independent, predefined bacterial species. Gram-stain reaction, colony and cell morphology, biochemical characteristics, isolation source, and NIBR IDs are described in the species description section.

• Korean Journal of Veterinary Research
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• v.61 no.1
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• pp.2.1-2.6
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• 2021

The reproductive tracts have an intimate relationship with reproduction because there are bacterial communities that can affect reproductive health. The differences in the bacterial community of periparturient dairy cows were investigated. Vaginal and fecal samples were collected seven days before and after calving, and DNA was extracted to sequence the V3-V4 regions of the 16S rRNA genes. In the postpartum vaginas, operational taxonomic units, Chao1, Shannon, and Simpson were decreased, and phyla Fusobacteria and Bacteroidetes were increased. In summary, bacterial abundance can affect the periparturient biological differences in dairy cows, suggesting a susceptibility to infection within one week after calving.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.4
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• pp.563-569
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• 2013

This study was carried out to investigate the dose-response of chitooligosaccharide (with a molecular weight of 1~3 kDa) on antimicrobial activity and lipid lowering functions in rats. Sprague-Dawley male rats were given experimental diets containing 0 (control), 0.5, 2, or 5% chitooligosaccharide (COS) for 5 weeks. Weight gain and food intake were significantly lower in rats fed 5% COS than control rats and rats fed 0.5 and 2% COS. The numbers of fecal bacteria, including bifidobacteria, lactobacilli, bacteroides, total anaerobes, and total aerobes, which reflect gut microbiota, were significantly decreased in rats fed 5% COS. Plasma triglyceride concentrations significantly decreased in a dose-dependent manner in rats fed 2% or 5% COS, while plasma total cholesterol was not significantly different among groups. The hepatic concentration of triglycerides was lower in rats fed 5% COS, and fecal triglycerides significantly increased in rats fed 5% COS. These results indicate that 5% COS supplementation in a diet may exert antimicrobial activity in vivo, and inhibit the proliferation of typical gut microbes, while lowering lipids.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.10
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• pp.1660-1669
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• 2018

Objective: This study was conducted to evaluate the effect of probiotics (Bacillus subtilis and Enterococcus faecium) and xylo-oligosaccharide (XOS) supplementation on growth performance, nutrient digestibility, serum profiles, intestinal health, fecal microbiota and noxious gas emission in weanling pigs. Methods: A total of 240 weanling pigs ([Yorkshire${\times}$Landrace]${\times}$Duroc) with an average body weight (BW) of $6.3{\pm}0.15kg$ were used in this 28-day trial. Pigs were randomly allocated in 1 of the following 4 dietary treatments in a $2{\times}2$ factorial arrangement with 2 levels of probiotics (0 and 500 mg/kg probiotics) and XOS (0 and 200 mg/kg XOS) based on the BW and sex. Results: Administration of probiotics or XOS improved average daily gain (p<0.05) during 0 to 14 d and the overall period, while pigs that were treated with XOS had a greater average daily gain and feed efficiency (p<0.05) compared with unsupplemented treatments throughout 15 to 28 d and the whole experiment. Either probiotics or XOS treatments increased the apparent total tract digestibility of nutrients (p<0.05) during 0 to 14 d. No effects on serum profiles were observed among treatments. The XOS increased villus height: crypt depth ratio in jejunum (p<0.05). The supplementation of probiotics (500 mg/kg) or XOS (200 mg/kg) alone improved the apparent total tract digestibility of dry matter, nitrogen and gross energy on d 14, the activity of trypsin and decreased fecal NH3 concentration (p<0.05). Administration of XOS decreased fecal Escherichia coli counts (p<0.05), while increased lactobacilli (p<0.05) on d 14. There was no interaction between dietary supplementation of probiotics and XOS. Conclusion: Inclusion of XOS at 200 mg/kg or probiotics (Bacillus subtilis and Enterococcus faecium) at 500 mg/kg in diets containing no antibiotics significantly improved the growth performance of weanling pigs. Once XOS is supplemented, further providing of probiotics is not needed since it exerts little additional effects.

• 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.