Decoding the intestinal microbiota repertoire of sow and weaned pigs using culturomic and metagenomic approaches

( Daye Mun ),( Hayoung Kim ),( Minhye Shin ),( Sangdon Ryu ),( Minho Song ),( Sangnam Oh ),( Younghoon Kim ) 한국축산학회 2021 한국축산학회지 Vol.63 No.6

To elucidate the role and mechanism of microbes, we combined culture-dependent and culture-independent approaches to investigate differences in gut bacterial composition between sows and weaned pigs. Under anaerobic conditions, several nonselective and selective media were used for isolation from fecal samples. All isolated bacteria were identified and classified through 16S rRNA sequencing, and the microbiota composition of the fecal samples was analyzed by metagenomics using next generation sequencing (NGS) technology. A total of 278 and 149 colonies were acquired from the sow and weaned pig fecal samples, respectively. Culturomics analysis revealed that diverse bacterial genus and species belonged to Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes were isolated from sow and weaned pigs. When comparing culture-dependent and culture-independent analyses, 191 bacterial species and 2 archaeal bacterial species were detected through culture-independent analysis, and a total of 23 bacteria were isolated through a culture-dependent approach, of which 65% were not detected by metagenomics. In conclusion, culturomics and metagenomics should be properly combined to fully understand the intestinal microbiota, and livestock-derived microbial resources should be informed by culturomic approaches to understand and utilize the mechanism of host-microbe interactions.

Effects of Bacillus-based probiotics on growth performance, nutrient digestibility, and intestinal health of weaned pigs

( Daye Mun ),( Hyunjin Kyoung ),( Myunghwan Kong ),( Sangdon Ryu ),( Ki Beom Jang ),( Jangryeol Baek ),( Kyeong Il Park ),( Minho Song ),( Younghoon Kim ) 한국축산학회 2021 한국축산학회지 Vol.63 No.6

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.

Perspectives on Bovine Milk-Derived Extracellular Vesicles for Therapeutic Applications in Gut Health

Daye Mun,Sangnam Oh,Younghoon Kim 한국축산식품학회 2022 한국축산식품학회지 Vol.42 No.1

Extracellular vesicles (EVs) are nanosized vesicles secreted from cells into the extracellular environment and are composed of a lipid bilayer that contains cargos with biological activity;such as lipids;proteins;mRNAs;and noncoding microRNAs (miRNAs). Due to their biological activity and their role in cell-to-cell communication;interest in EVs is rapidly increasing. Bovine milk is a food consumed by people of all ages around the world that contains not only a significant amount of nutrients but also EVs. Milk-derived EVs also exhibit biological activity similar to other source-derived EVs;and studies on bovine milk EVs have been conducted in various research fields regarding sufficient milk production. In particular;not only are the effects of milk EVs themselves being studied;but the possibility of using them as drug carriers or biomarkers is also being studied. In this review;the characteristics and cargo of milk EVs are summarized;as well as their uptake and stability;efficacy and biological effects as carriers;and future research directions are presented.

Value of clay as a supplement to swine diets

Daye Mun,Jongmoon Lee,Jeehwan Choe,Byeonghyeon Kim,Sangnam Oh,Minho Song 충남대학교 농업과학연구소 2017 Korean Journal of Agricultural Science Vol.44 No.2

Spore-forming probiotic bacterium Bacillus coagulans influences on the flavor of Gouda cheese using micro-cheese platform

Hyejin Choi,Sangdon Ryu,Daye Mun,Woongji Lee,An Na Kang,Mingeun Kang,Hayoung Kim,Daniel Lee,Sangnam Oh,Younghoon Kim 한국식품영양과학회 2021 한국식품영양과학회 학술대회발표집 Vol.2021 No.10

Bacillus coagulans is a gram-positive, spore-forming and potential probiotic bacterium in food industry. Spore-forming bacteria is enable to survive in the host"s gastrointestinal tract functioning as probiotics that could play a beneficial role in the host health. We evaluated the probiotic characteristics including acid, bile and heat tolerance of B. coagulans SRCM121381 isolated from human breast milk. SRCM121381 showed high resistance to acid and bile, heat tolerance. We employed FIMM (Fermentor for Intestine Microbiota Model) system to evaluate effects of SRCM 121381 on the microbial dynamics in gut environments. SRCM121381 resulted in increased population of Romboutsia and Turicibacter at the genus level, while the abundance of Escherichia decreased significantly. We added SRCM 121381 with starter strains for Gouda cheese using micro-cheese platform and evaluated on the change of cheese flavor. SRCM121381 increased acetic acid (Vinegar) and decreased esters and ketone as undesirable flavor such as bitter taste. Our results propose that potential probiotic bacterium B. coagulans SRCM121381 could be applied on dairy foods as an adjunct starter with flavor productivity.

Short Communication: Molecular characterization and functionality of rumen-derived extracellular vesicles using a Caenorhabditis elegans animal model

Hye Jin Choi,Daye Mun,류상돈,Min-Jin Kwak,Bum-Keun Kim,Dong-Jun Park,Sangnam Oh,Young Hoon Kim 한국축산학회 2023 한국축산학회지 Vol.65 No.3

The rumen fluids contain a wide range of bacteria, protozoa, fungi, and viruses. The various ruminal microorganisms in the rumen provide nutrients by fermenting the forage they eat. During metabolic processes, microorganisms present in the rumen release diverse vesicles during the fermentation process. Therefore, in this study, we confirmed the function of rumen extracellular vesicles (EVs) and their interaction with the host. We confirmed the structure of the rumen EVs by transmission electron microscope (TEM) and the size of the particles using nanoparticle tracking analysis (NTA). Rumen EVs range in size from 100 nm to 400 nm and are composed of microvesicles, microparticles, and ectosomes. Using the Caenorhabditis elegans smart animal model, we verified the interaction between the host and rumen EVs. Exposure of C. elegans to rumen EVs did not significantly enhance longevity, whereas exposure to the pathogenic bacteria Escherichia coli O157:H7 and Staphylococcus aureus significantly increased lifespan. Furthermore, transcriptome analysis showed gene expression alterations in C. elegans exposed to rumen EVs, with significant changes in the metabolic pathway, fatty acid degradation, and biosynthesis of cofactors. Our study describes the effect of rumen EV interactions with the host and provides novel insights for discovering biotherapeutic agents in the animal industry.

Effects of a probiotic Lactobacillus strain on behavioral response and feces metabolite content during chronic mild stress in mice

Jae Gwang Song,Bomi Lee,Daye Mun,Younghun Kim,Minjee Lee,Jungwoo Yang,Hyung Wook Kim 한국실험동물학회 2023 한국실험동물학회 학술발표대회 논문집 Vol.2023 No.2

Enhanced γ-aminobutyric acid and sialic acid in fermented deer antler velvet and immune promoting effects

( Jiseon Yoo ),( Juyeon Lee ),( Ming Zhang ),( Daye Mun ),( Minkyoung Kang ),( Bohyun Yun ),( Yong-an Kim ),( Sooah Kim ),( Sangnam Oh ) 한국축산학회 2022 한국축산학회지 Vol.64 No.1

Deer antler velvet is widely used in traditional medicine for its anti-aging, antioxidant, and immunity-enhancing effects. However, few studies have reported on the discovery of probiotic strains for deer antler fermentation to increase functional ingredient absorption. This study evaluated the ability of probiotic lactic acid bacteria to enhance the concentrations of bioactive molecules (e.g., sialic acid and gamma-aminobutyric acid [GABA]) in extracts of deer antler velvet. Seventeen strains of Lactobacillus spp. that were isolated from kimchi and infant feces, including L. sakei, L. rhamnosus, L. brevis, and L. plantarum, and those that improved the life span of Caenorhabditis elegans were selected for evaluation. Of the 17 strains, 2 (L. rhamnosus LFR20-004 and L. sakei LFR20-007) were selected based on data showing that these strains increased both the sialic acid and GABA contents of deer antler extract after fermentation for 2 d and significantly improved the life span of C. elegans. Co-fermentation with both strains further increased the concentrations of sialic acid, GABA, and metabolites such as short-chain fatty acids and amino acids. We evaluated the biological effects of the fermented antler velvet (FAV) on the antibacterial immune response in C. elegans by assessing worm survival after pathogen infection. The survival of the C. elegans conditioned with FAV for 24 h was significantly higher compared with that of the control worm group fed only normal feed (non-pathogenic E. coli OP50) exposed to E. coli O157:H7, Salmonella typhi, and Listeria monocytogenes. To evaluate the protective effects of FAV on immune response, cyclophosphamide (Cy), an immune-suppressing agent was treated to in vitro and in vivo. We found that FAV significantly restored viability of mice splenocytes and immune promoting-related cytokines (interleukin [IL]-6, IL-10, inducible nitric oxide synthase [iNOS], interferon [IFN]-γ, and tumor necrosis factor [TNF]-α) were activated compared to non-fermented deer antlers. This finding indicated the protective effect of FAV against Cy-induced cell death and immunosuppressed mice. Taken together, our study suggests that immune-promoting antler velvet can be produced through fermentation using L. rhamnosus LFR20-004 and L. sakei LFR20-007.

The Ingestion of Dietary Prebiotic Alternatives during Lactation Promotes Intestinal Health by Modulation of Gut Microbiota

Ryu Sangdon,Lee Jeong Jae,Mun Daye,김수린,Choe Jeehwan,Song Minho,Kim Younghoon 한국미생물·생명공학회 2022 Journal of microbiology and biotechnology Vol.32 No.11

Palm kernel expeller (PKE), a by-product of palm oil extraction, contains higher amounts of fiber than corn and soybean meal, but offers low energy density, protein value, and amino acid (AA) composition, limiting its use for swine. Recently however, it was reported that dietary fiber has a positive effect on the gut microbiota of the host, and therefore it is necessary to study the effect of PKE feeding on the intestinal microbiota of swine. In this study, we investigated the effects of supplementation with PKE in lactation diets on the gut microbiota composition of lactating sows and their litters. A total of 12 sows were randomly assigned to two dietary treatment groups in a completely randomized design. The treatments were a diet based on corn-soybean meal (CON) and CON supplemented with 20% of PKE. Sow and piglet fecal samples were collected before farrowing, on days 7 and 28 (weaning) after farrowing, and on days 7 and 28 (weaning) after farrowing, respectively, to verify gut microbiota composition by pyrosequencing analysis. The beta-diversity result showed a significant difference only in weaning-stage piglets, but dietary PKE altered the gut microbiota in sows by increasing the abundance of Lactobacillus compared with CON. In piglets, dietary PKE decreased the abundance of opportunistic pathogen Proteus and increased the abundance of potentially beneficial bacteria, such as Prevotellaceae and Prevotella. Our results can be helpful in developing feeding strategies and support the beneficial effects of dietary PKE to improve the gut health of animals.