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The intestinal microbiota, an invisible organ supporting a host’s survival, has essential roles in metabolism, immunity, growth, and development. Due to the influence of the intestinal microbiota on the biology of an organism, application of such data to wildlife conservation has gained interest. While there is a standard protocol for studying the human intestinal microbiota, an equivalent one for wildlife is still under development. A major challenge is sampling the intestinal microbiota in an effective, unbiased way. Faecal samples are a popular proxy of the intestinal microbiota because collection is non-invasive, convenient, and allows for longitudinal sampling. Yet, it is unclear whether the faecal microbiota is representative of the intestinal microbiota. In amphibians, research on sampling methodology is limited. In this study, we first characterise and compare microbiota (small intestine, large intestine, faeces) of two Hong Kong stream-dwelling frog species: the Lesser Spiny Frog (Quasipaa exilispinosa) and the Hong Kong Cascade Frog (Amolops hongkongensis). We found that both species have similar dominant phyla and families, but diverge in terms of the dominant genera. Second, we assess the performance of faecal microbiota in representing the intestinal microbiota in these two species. By comparing the microbiota between three sample types (small intestine, large intestine, and faeces), we found (1) that microbiota of small and large intestine differs significantly, (2) that faeces are not an appropriate proxy of both intestinal sections, and (3) a set of microbial taxa that significantly differ between sample type. Our study raises caution on equating faecal and intestinal microbiota. The use of faeces can avoid sacrifice an animal, but researchers should avoid over-extrapolation and interpret results carefully.

Recently, we reported the robust in vitro three-dimensional (3D) expansion of intestinal organoids derived from adult bovine (> 24 months) samples. The present study aimed to establish an in vitro 3D system for the cultivation of intestinal organoids derived from growing cattle (12 months old) for practical use as a potential alternative to in vivo systems for various purposes. However, very few studies on the functional characterization and 3D expansion of adult stem cells from livestock species compared to those from other species are available. In this study, intestinal crypts, including intestinal stem cells, from the small intestines (ileum and jejunum) of growing cattle were isolated and long-term 3D cultures were successfully established using a scaffold-based method. Furthermore, we generated an apical-out intestinal organoid derived from growing cattle. Interestingly, intestinal organoids derived from the ileum, but not the jejunum, could be expanded without losing the ability to recapitulate crypts, and these organoids specifically expressed several specific markers of intestinal stem cells and the intestinal epithelium. Furthermore, these organoids exhibited key functionality with regard to high permeability for compounds up to 4 kDa in size (e.g., fluorescein isothiocyanate [FITC]-dextran), indicating that apical-out intestinal organoids are better than other models. Collectively, these results indicate the establishment of growing cattle-derived intestinal organoids and subsequent generation of apical-out intestinal organoids. These organoids may be valuable tools and potential alternatives to in vivo systems for examining host-pathogen interactions involving epithelial cells, such as enteric virus infection and nutrient absorption, and may be used for various purposes.

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.

Wheat gluten is an increasingly common ingredient in poultry diets but its impact on the small intestine in chicken is not fully understood. This study aimed to identify effects of high-gluten diets on chicken small intestines and the variation of their associated transcriptional responses by age. A total of 120 broilers (Ross Strain) were used to perform two animal experiments consisting of two gluten inclusion levels (0% or 25%) by bird’s age (1 week or 4 weeks). Transcriptomics and histochemical techniques were employed to study the effect of gluten on their duodenal mucosa using randomly selected 12 broilers (3 chicks per group). A reduction in feed intake and body weight gain was found in the broilers fed a high-gluten containing diet at both ages. Histochemical photomicrographs showed a reduced villus height to crypt depth ratio in the duodenum of gluten-fed broilers at 1 week. We found mainly a significant effect on the gene expression of duodenal mucosa in gluten-fed broilers at 1 week (289 differentially expressed genes [DEGs]). Pathway analyses revealed that the significant DEGs were mainly involved in ribosome, oxidative phosphorylation, and peroxisome proliferator-activated receptor (PPAR) signaling pathways. These pathways are involved in ribosome protein biogenesis, oxidative phosphorylation and fatty acid metabolism, respectively. Our results suggest a pattern of differential gene expression in these pathways that can be linked to chronic inflammation, suppression of cell proliferation, cell cycle arrest and apoptosis. And via such a mode of action, high-gluten inclusion levels in poultry diets could lead to the observed retardation of villi development in the duodenal mucosa of young broiler chicken.

Four Korean native steers (511 ± 17.2 kg; 2 × 2 replicated crossover design) fitted with duodenal cannulas were used to investigate the influence of oral administration of soluble whey protein (WP; 82.29% crude protein) on ruminal fermentation, gastrointestinal (GI) hormone secretion in the blood, pancreatic α-amylase activity in the duodenum, and disappearance rate in each segment of the GI tract. Steers were orally fed the basal diet (control; TMR [total mixed ration] 9 kg/d) or the basal diet with enriched WP (400 g/d) for 14 days. The apparent crude protein disappearance rate in the rumen of the WP was higher than in control (p < 0.05). However, no difference between groups was observed in the apparent crude protein disappearance rate in the intestine and the apparent starch disappearance rates in the rumen, GI tract. The level of cholecystokinin, secretin, and ghrelin in serum and pancreatic α-amylase activity in the duodenum of the WP also did not change. The changes in the level of blood urea nitrogen related to protein metabolism were higher in the WP than in the control (p < 0.05). However, the levels of total protein, lipid, carbohydrate and mineral metabolites did not change. Consequently, we suggest that the oral administration of WP in steers assisted in ruminal fermentation due to the population increase of microbes in the rumen but did not improve the starch digestion rate in the small intestine because GI hormone secretion in the blood and pancreatic α-amylase activity did not change.

This study aimed to confirm the applicability of the new nitrogen (N2) gas stunning method in the broiler slaughtering process by comparing the meat and small intestine quality following different stunning methods (electrical, carbon dioxide (CO2), N2, and halal). Four treatments were compared: (i) electrical stunning (Elec), (ii) 80% CO2 gas stunning (CO2-gas), (iii) 98% N2 gas stunning (N2-gas), and (iv) the non-stunning method (halal). N2 gas stunning (98%) and the halal method were conducted at the pilot plant abattoir of the national institute of animal science, Korea, and electrical and 80% CO2 stunning were performed on the nearest commercial slaughter house. Meat pH24h, color (lightness, redness and yellowness), proximate composition, water holding capacity (WHC), cooking loss, and Warner-Bratzler shear force (WBSF) were measured, and in the small intestine, pH24h, color, thickness, and WBSF were measured. The Elec treatment showed high lightness, yellowness, and low redness in both meat and the small intestine, indicated by a pale color; the CO2-gas treatment showed high redness, low lightness, and low yellowness, and the coloration of meat from the N2-gas treatment was intermediate between Elec and CO2-gas. For other quality traits, the N2-gas showed good results and was between Elec and CO2-gas. Additionally, severe stress (low pH in both meats), low WHC in meat, and cracked small intestine with numerous apertures were observed in the CO2-gas, and pale colored hemorrhagic breast meat was found in the Elec. Therefore, in view of animal welfare and quality traits of meat and the small intestine, 98% N2 gas can be considered in broiler stunning.

In the published article “Impacts of whey protein on starch digestionin rumen and small intestine of steers. J Anim Sci Technol. 2019;61(2):98-108. https://doi.org/10.5187/jast.2019.61.2.98,” the4th author’s name and affiliation are given incorrectly [1]. The editorialoffice would like to correct the author’s name and affiliation. Corrected name and affiliation of 4th author :Kyoung-Hoon Kim44Graduate School of International Agricultural Technology, PyeongchangCampus, Seoul National University, Pyeongchang 25354,Korea

항생제 대체물질로서 가축용 생균제를 개발하기 위하여, 오리의 소장 및 맹장으로부터 416주의 미생물을 분리하였으며, 항균물질인 박테리오신을 생산하는 13주를 최종적으로 선발하였다. 선발균주들은 Lactobacillus salivarius 1주, Lactobacillus plantarum 1주, Pediococcus pentosaceus 1주, Enterococcus 7주, 그리고 3주의 E. coli로 동정되었으며, 복합생균제를 제조할 목적으로 상기 균주중 5균주에 대한 박테리오신 및 생균제적인 특성를 조사하였다. 유산균인 Lact. salivarius JWS 58, Ent. faecium JWS 833, 그리고 Ped. pentosaceus JWS 939는 식품관련 병원성 균주로 알려진 Listeria mono- cytogenes에 대하여 강한 항균능력을 나타냈으며, 상기 균주의 cell-free supernatant를 단백질 분해효소 처리시 대부분이 병원균 억제활성을 상실하였다. 선발균주인 E. coli JWS 108은 그람음성균주인 E. coli와 Staphylococcus aureus 균주에 대한 생육억제활성을 나타냈다. 또한 Lact. salivarius JWS 58균주와 Ent. faecium JWS 833균주는 각각 Lactobacillus sp. 및 Enterococcus sp. 균주에 대하여 억제활성을 지니고 있었다. Lact. salivarius JWS 58균주는 pH 2.5 완충용액에서 2시간 동안 50% 이상 생존하였으며, pH 2.0 이하에서는 대부분의 선발균주들이 사멸하였다. 그러나 선발균주 모두 0.5% 담즙산 농도까지 아무런 억제현상없이 정상적으로 성장하였다. Ped. pentosaceus JWS 939와 Lact. plantarum JWS 1354 균주는 상대적으로 높은 amylase와 cellulase 효소 활성를 나타냈으며, E. coli JWS 108 균주는 낮은 효소활성을 보였다. Protease 및 lipase 효소 활성은 5균주 모두 유사하게 나타내었다.

The aim of this study was to isolate and characterize bacteriocin-producing bacteria from the intestine of duck to use as probiotics for livestock. A total of 416 strains were isolated from the small intestine and cecum of ducks and 13 isolates were finally selected after determinging inhibitory activity against pathogenic indicators by spot-on-lawn method. The selected strains were identified as Lactobacillus salivarius JWS 58, Lactobacillus plantarum JWS 1354, Pediococcus pentosaceus JWS 939, 7 strains of enterococci, and 3 strains of Escherichia coli. Lact. salivarius JWS 58, Ent. faecium JWS 833, and Ped. pentosaceus JWS 939 showed a strong inhibitory activity against Listeria monocytogenes. E. coli JWS 108 inhibited the growth of E. coli and Staphylococcus aureus. Lact. salivarius JWS 58 strain survived almost 50% in pH 2.5 phosphate buffer for 2 hr. Ped. pentosaceus JWS 939 and Lact. plantarum JWS 1354 showed strong amylolytic activity. These results suggest that a combination of bacteriocins or multispecies probiotics of the selected strains has a strong potential of alternative to antibiotics in livestock production.

The aim of this study is to develop an in vitro intestinal system for studying drug absorption and microbial interaction. Human intestinal epithelial cell line (Caco-2) cultured on a petri dish cannot properly reconstitute human intestine because the former is different from the latter in terms of physiological and morphological properties such as epithelial electrical resistance and production of apical brush amino peptidase enzyme.In this presentation, we show that this discrepancy can be reduced by growing Caco-2 cells in microfluidic cell culture device (μFCCD) where a polyester membrane is installed between two PDMS (polydimethylsiloxane) layers having microchannels perfusing nutrients. Cells in μFCCD were taller than those in a petri dish, while F-actin in the apical brush border of the former was denser than that in the apical brush border of the latter. The former formed villi-like structures, which were not well observed in those in a petri dish. These results suggest that the microenviroment in μFCCD mimics the in vivo tissue microenvironment and the intestinal cells in the chip can be ready for studying drug absorption as well as host-bacteria interaction.

In this study, the effects of processed small black soybeans on the intestinal morphological characteristics of ratswere examined. Adult male rats were fed diets containing raw, cooked, or acidified small black soybean powders for 4 weeks.The total short chain fatty acid (SCFA) level was significantly higher in the acidified small black soybean supplemented groupthan in the raw and cooked soybean diet groups. The major SCFAs found in the experimental groups were acetate, followedby propionate and butyrate. The duodenal villus height and colonic mucosal thickness were also significantly higher in theacidified small black soybean supplemented group than in the raw and cooked soybean diet groups. The acidified small blacksoybean supplemented group showed the densest colonic mucosa by staining with alcian blue (AB), as compared to the rawand cooked soybean diet groups. The acidified small black soybean supplemented group exhibited strongly stained CD4+ inthe mucosal lamina propria, while cooked and acidified diet groups were more strongly stained CD8+ in the submucosallamina propria than the raw diet group. These results suggest that acidified small black soybeans may help improve intestinalfunction.

We investigated the effects of acupuncture on proximal colonic motility in conscious dogs. Two of stainlesssteel electrodes were implanted at the proximal colon in 5 healthy mongrel dogs. Manual acupuncture was applied withoutadditional stimulation at the following acupoints, LI01, LI02, LI03, LI04, LI05, LI06, LI07 or LI11. All dogs exhibitedspontaneous colonic activity. Manual acupuncture at LI06 depressed colonic motility by decreasing the total duration andthe frequency and increasing the cycle length of contractile states. While LI07 inhibited only the total duration, LI03 acu-point significantly depressed the cycle length without affecting other factors of contractile states. These findings dem-onstrated that acupuncture could have specific effects on colonic motility in different ways depending on acupoints.acupuncture at LI06 or LI07 decreasing colonic motility may be used for the treatment of diseases caused by an increasedcolonic motility, and acupuncture at LI03 reducing the cycle length of colonic motility may be useful for constipation. Acu-points of large intestine meridian have point-specific effects on proximal colonic motility in conscious dogs.

The research aimed to identify the lacted acid bacteria from intestinal of milkfish (Chanos chanos) potential acvity pathogen anbacteria used PCR 18s Rrna Methode. The lactic acid bacteria were isolated using selective media MRS agar for lactic acid bacteria. Identification isolates PCR 18S rRNA and sequencing method that are DNA isolation, DNA amplification use PCR, DNA visualizasion from amplification with electroforesis, Basic Local Alignment Search Tool (BLAST) and Tree view program. In view of PCR analysis and tree view program, LAB is Lactobacillus acidophilus strain ATTC 4796.

We show here that polymeric nanoparticles with sizes of 100 nm and under can be dispersed in the body of nematode C. elegans through the receptor-mediated endocytosis. When we fed animals with 50, 100, 200, and 500 nm polystyrene clusters, only 50 and 100 nm particles could be delivered to internal organs from intestine. By the loss-offunction mutations in genes related with endocytosis, we found that nanoparticles could be transferred from intestine to neighbor organs through receptor-mediated endocytosis.

To produce a novel bacteriocin against fish pathogenic bacteria, Lactococcus lactis subsp. was isolated from flounders collected from aquaculture farms. The antimicrobial activity and some physico-chemical properties of the purified bacteriocin were studied. The bacteriocin was purified by 15% PEG 4000/20% Na2SO4 and ethanol saturation followed by twice C-18 reverse phase HPLC. The molecular weight of the bacteriocin measured by MALDI-TOF was 3.3 kDa. We found that the bacteriocin inhibited the growth of gram-positive bacteria including fish pathogenic bacteria, Streptococcus sp. and Lactococcus garviae, but not the growth of gram-negative bacteria. Especially, S. pleomorphus indicator strain (400 arbitrary units ml-1), which is a harmful pathogenic bacteria to aquaculture fish, was shown to be most sensitive . The bacteriocin was found to be stable at 100°C for 20 min and in pH ranged from 3.0-4.0, but it was unstable at pH 10.0. The results indicates that antimicrobial agent produced by L. lactis subsp. may be a good candidate instead of antibiotics to apply in aquaculture and seafood industries.