• Journal of Animal Science and Technology
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• 제64권6호
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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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• 제27권11호
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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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• 제39권2호
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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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• 제63권6호
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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.

• 한국가금학회지
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• 제49권2호
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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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• 제9권1호
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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.

• 대한수의학회지
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• 제61권1호
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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.

• 한국식품영양과학회지
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• 제42권4호
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• pp.563-569
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• 2013

본 연구는 흰쥐에서 저분자량(1~3 kDa) 키토올리고당(chitooligosaccharide, COS)이 장내 미생물에 미치는 항균활성과 지질 개선에 관한 용량-반응 효과를 조사하기 위하여 수행되었다. 4주령 된 Sprague-Dawley 수컷 32마리를 1주일 동안 사육 환경에 적응시킨 후 군당 8마리씩 4군으로 나누어서 고지방식이에 키토올리고당을 0%(대조식이), 0.5%, 2%, 5% 수준으로 첨가한 식이를 5주 동안 자유롭게 급여하였다. 실험식이 5주째에 장 통과시간을 측정하였고, 신선한 분변을 취하여 장내 균수를 측정하였으며, 희생 후 혈장 지질, 간 지질, GPT 활성, 분변 지질을 측정하였다. 체중 증가량과 식이섭취량은 COS 5% 첨가군에서 유의하게 낮았다. 분변 중 bifidobacteria, lactobacilli, bacteroides, total anaerobes, total aerobes 수는 대조군, COS 0.5% 첨가군과 COS 2% 첨가군 사이에 차이가 없었으나, COS 5% 첨가군에서는 모두 유의하게 감소하였다. 혈장 총 콜레스테롤, HDL-C 및 LDL-C 농도는 군간 차이가 없었으나, 중성지방 농도는 COS 2% 첨가군과 COS 5% 첨가군에서 유의하게 낮았으며, 키토올리고당의 용량이 높아질수록 낮았다. 간 조직의 중성지방 농도는 COS 5% 첨가군에서 낮았으며, 반면에 분변의 콜레스테롤 및 중성지방 농도는 COS 5% 첨가군에서 높았으나 담즙산은 차이가 없었다. 결론적으로 저분자량(1~3 kDa) 키토올리고당을 5% 첨가한 식이는 흰쥐의 식이섭취량, 체중증가량, 혈장 중성지방을 낮추어 지질 개선효과를 보였고, 분변의 혐기성균 중에 bacteroides의 균수를 크게 감소시켰으며 bifidobacteria와 lactobacilli 균수도 억제함으로써 장 미생물생태 변화를 초래하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권10호
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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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• 제53권4호
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• pp.390-405
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• 2020

본 연구는 probiotics와 prebiotics로 구성된 synbiotics 섭취가 건강한 성인의 배변 활동 및 변내 균총의 개선 효과를 확인하고자 하였고, probiotics 섭취 중단 후에도 prebiotics인 XOS 섭취가 장내 유익균총의 유지에 미치는 영향을 파악함으로써 XOS의 건강기능성 확인을 위한 근거자료를 확보하고자 하였다. 본 연구결과 BL2XO6군과 BL2XO2군의 체위 자료는 두 군 간에 유의적인 차이를 보이지 않았다. 변내 pH는 시료 섭취 전 두 군 모두 7.0이하로 약 산성이었으나, 시료 섭취 1주차 BL2XO6군에서 pH 7.16으로 유의적으로 높았으나 2주차 이후는 pH 7.0 이하의 값을 보여 시료 섭취에 따른 차이는 나타나지 않았다. 변내 균총의 변화를 보면 BL2XO6군의 경우 Clostridium spp. 은 시료 섭취 후 변 내 균 수가 감소하는 양상을 보인 반면, Bifidobacterium spp.은 시료 섭취 후 서서히 증가하는 양상을 보여 4주차에 시료 섭취 전에 비해 유의적으로 높았으나, 이외 주차에서는 유의적인 차이를 보이지 않았다. B. longum은 모든 주차에서 통계적으로 유의하지 않았으나 서서히 증가하는 추이를 보였다. BL2XO2군의 경우 시료 섭취 후 Lactobacillus spp.과 B. longum은 synbiotics 섭취 1주에만 증가하는 추이를 보였다. 각 주차별로 변 내 균총의 변화율에서 군별로 유의적인 차이를 보인 균총은 Clostridium spp.와 Lactobacillus spp.이었다. 즉, Clostridium spp.은 2주차에서, Lactobacillus spp.는 4주차에서 BL2XO6군과 BL2XO2군 사이에서 유의적인 차이를 보였다. 시료 섭취 전·후 Bacteroidetes와 Firmicutes의 비율은 두 군 모두 변화가 없는 것으로 나타났다. 또한, 시료 섭취 전·후의 변의 양, 변의 상태 및 배변 횟수, 복통 및 CSS점수에 유의적인 차이를 보이지 않았을 뿐만 아니라 군에 따른 차이 역시 존재하지 않았다. 이상의 결과 synbiotics 섭취 후 prebiotics만을 섭취할지라도 장내 균총의 변화에 긍정적인 영향을 미치는 것으로 나타나고 있을 뿐만이 아니라 정상적인 배변 활동을 유지하고 있어 synbiotics 섭취 후 prebiotics로 XOS 섭취는 자일로올리고당을 선택적으로 이용할 수 있는 B. longum을 장내에서 지속적으로 증식할 수 있도록 도와 장내 균총을 개선하는 효과와 정상적인 배변활동을 통한 장 건강에 도움을 줄 수 있을 것으로 사료된다.