Background: Aflatoxin B1 (AFB1) is a toxic metabolite generated by Aspergillus species and is commonly detected during the processing and storage of food; it is considered a group I carcinogen. The hepatotoxic effects, diseases, and mechanisms induced by AFB1 owing to chronic or acute exposure are well documented; however, there is a lack of research on its effects on the intestine, which is a crucial organ in the digestive process. Dogs are often susceptible to chronic AFB1 exposure owing to lack of variation in their diet, unlike humans, thereby rendering them prone to its effects. Therefore, we investigated the effects of AFB1 on canine small intestinal epithelial primary cells (CSIc). Methods: We treated CSIc with various concentrations of AFB1 (0, 1.25, 2.5, 5, 10, 20, 40, and 80 μM) for 24 h and analyzed cell viability and transepithelial-transendothelial electrical resistance (TEER) value. Additionally, we analyzed the mRNA expression of tight junction-related genes (OCLN, CLDN3, TJP1, and MUC2), antioxidant-related genes (CAT and GPX1), and apoptosis-related genes (BCL2, Bax, and TP53). Results: We found a significant decrease in CSIc viability and TEER values after treatment with AFB1 at concentrations of 20 μM or higher. Quantitative polymerase chain reaction analysis indicated a downregulation of OCLN, CLDN3, and TJP1 in CSIc treated with 20 μM or higher concentrations of AFB1. Additionally, AFB1 treatment downregulated CAT , GPX1, and BCL2. Conclusions: Acute exposure of CSIc to AFB1 induces toxicity, and exposure to AFB1 above a certain threshold compromises the barrier integrity of CSIc.
Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.
Purpose: This study was conducted to assess the post-discharge experiences of caregiving mothers of pediatric patients with intestinal failure who were receiving home TPN treatment. Methods: This was a qualitative study utilizing Colaizzi’s phenomenological research method. The eight participants were mothers of pediatric outpatients from the short-gut syndrome clinic at a tertiary hospital in Seoul who were continuing home TPN treatment through a CVC. Data were collected from January to May 2022 through individual in-depth interviews, and analyzed. Results: Analysis of 127 meaningful statements from the mothers identified 12 themes and 36 sub-themes, organized into five categories: “Mixed emotions regarding hospital discharge”, “Problems after discharge”, “Stress in everyday life”, “Support from nurses and family members”, and “Looking to the future”. Conclusion: The study results provided insights into the meaning and value of the post-discharge experiences of mothers of pediatric patients with intestinal failure. These findings will be valuable in the development of interventions to provide education and other support measures for primary caregivers of children with intestinal failure.
To produce an intestinal immunomodulatory beverage containing Centella asiatica extract (CAE), three types of CAE-added beverage prototypes were prepared, and their immunomodulatory activities and marker compounds were analyzed. As a result of the cytotoxicity assessment, all the beverages did not show significant toxicity compared to the control group. Next, the immunomodulatory activities of the beverage prototype were evaluated using the inflammatory model of IL-1β-induced intestinal epithelial cell line. All the samples significantly reduced the production of IL-6, IL-8, and MCP-1 in a CAE concentration-dependent manner. In addition, CAE-added beverages inhibited NO, IL-6, and IL-12 production in LPS-induced RAW 264.7 cells. When the major triterpenoids, as marker compounds for the production of CAE-added beverages, were analyzed by HPLC-DAD, only asiaticoside was detected beyond the limit of quantification, while madecassoside, madecassic acid, and asiatic acid were not detected. The amounts of asiaticoside in CAE-added beverage prototypes were confirmed in No. 1 (19.39 μg/mL), 2 (19.25 μg/mL), and 3 (19.98 μg/mL). In conclusion, the results of this study suggested that CAE-added beverage prototypes induced immunomodulatory effects in the intestinal inflammatory cell line models and asiaticoside could be used as a marker compound for CAE-added beverage production.
Microbes in insect gut significantly influence host physiology. While Lepidoptera is a diverse insect order, the relationship between microbial symbiosis and host development remains elusive, especially concerning role of gut-colonizing bacteria in metamorphosis. We investigated the gut microbial diversity in Galleria mellonella throughout its life cycle using 16S rRNA amplicon sequencing. Our findings revealed a predominance of Enterococcus spp. in larvae and Enterobacter spp. in pupae. Remarkably, removing Enterococcus spp. hastened the larval-to-pupal transition. Transcriptome analysis showed an upregulation of immune response genes in pupae and hormone genes in larvae. Notably, the production of antimicrobial peptides in the host gut varied with developmental stages. Some of these peptides suppressed the growth of Enterococcus innesii, a dominant gut bacterium in G. mellonella larvae. This research underscores the pivotal role of gut microbiota shifts in metamorphosis, driven by the secretion of antimicrobial peptides in the G. mellonella digestive system.
Prebiotics are known as components of intestinal microbiota that can improve and maintain human health status by stimulating the growth and activity of the intestinal tract as a method of controlling the intestinal environment. In this study, we examined whether 2’-fucosyllactose (FL) could affect intestinal microbial population and bowel activity. Water content and frequency of mouse feces were increased in the 2’-FL treated group at a high concentration (1,000 mg/kg), with brightness of the color enhanced and physical properties diluted. In addition, intestinal microbial analysis showed that harmful bacteria Clostridium and Staphylococcus strains were decreased and beneficial bacteria such as Lactobacillus strains were markedly increased in the group treated with a high concentration of 2’-FL compared to those in the control group. These findings suggest that administration of 2’-FL can maintain healthy bowel activity by reducing harmful bacteria population and improving diluted physical properties.
According to the classification of World Health Organization, primary adenocarcinomas of the sinonasal tract can be initially classified as salivary and non-salivary types. The latter are further divided into intestinal and non-intestinal types. Sinonasal intestinal-type adenocarcinoma(ITAC) is rare adenocarcinoma subtype, which is closely occupational exposure to hardwood dusts, leather. In this study, we present a case of ITAC in a 68-year-old man. We successfully treated with wide excision and soft tissue reconstruction with free anterolateral thigh flap.
This study was conducted to evaluate the effects of ImmunoSEB as an immune-booster additive on the performance of broiler chicks. A total of 1150 broiler chickens were randomly allotted to three treatments on the basis of body weight (10 pens per treatment with 40 broilers in each pen): the control (CON), CON + ImmunoSEB 0.025% in feed (SEB25), and CON + ImmunoSEB 0.050% in feed (SEB50). The experiment was conducted for d 42 in three phases (phase 1, d 0–14; phase 2, d 15–28; and phase 3, d 29–42). There were significant differences in the average daily gain (ADG) and feed intake. The ADG at d 14 in the SEB50 treatment was greater than that in the CON treatment. The overall ADG in the SEB50 treatment was greater than that in the CON treatment. During d 0–14, the feed intake of chickens in the SEB50 treatment increased compared to that in the CON treatment. The crude protein and lysine digestibility improved in the SEB25 and SEB50 treatments compared to those in the CON treatment at d 28. Superoxide dismutase concentration significantly increased in the SEB50 treatment compared to that in the CON treatment. The interleukin-1β and tumor necrosis factor-α concentrations were higher in the cecum of chickens in the CON treatment than in the SEB25 and SEB50 treatments. A lower population of E. coli was detected in the ileum and cecum of broilers fed the SEB50 diet compared to those of broilers fed the CON diet. The overall result showed the beneficial effects of using ImmunoSEB at a dose of 0.050% in broiler chickens.
Recent progress has been made to establish intestinal organoids for an in vitro model as a potential alternative to an in vivo system in animals. We previously reported a reliable method for the isolation of intestinal crypts from the small intestine and robust three-dimensional (3D) expansion of intestinal organoids (basal-out) in adult bovines. The present study aimed to establish next-generation intestinal organoids for practical applications in disease modeling-based host-pathogen interactions and feed efficiency measurements. In this study, we developed a rapid and convenient method for the efficient generation of intestinal organoids through the modulation of the Wnt signaling pathway and continuous apical-out intestinal organoids. Remarkably, the intestinal epithelium only takes 3-4 days to undergo CHIR (1 µM) treatment as a Wnt activator, which is much shorter than that required for spontaneous differentiation (7 days). Subsequently, we successfully established an apical-out bovine intestinal organoid culture system through suspension culture without Matrigel matrix, indicating an apical-out membrane on the surface. Collectively, these results demonstrate the efficient generation and next-generation of bovine intestinal organoids and will facilitate their potential use for various purposes, such as disease modeling, in the field of animal biotechnology.
Three-dimensional (3D) organoids act as model systems because they mimic in vivo tissue morphology. Recent advancements in the field have demonstrated that organoids derived from various organs have assisted in understanding the underlying mechanisms of disease modeling and expanded our knowledge of organ development in vitro. Furthermore, these organoids have become a promising biomaterial in regenerative medicine for therapeutic purposes as well as in nutritional research for feed efficiency measurement in livestock. Intestinal organoids of livestock, including pigs, cattle, chickens, and horses, have been developed. These could be used to examine host-pathogen interactions, such as interaction between enteric viruses and epithelial cells, and are potential alternatives to in vivo systems. However, there are very limited studies regarding species-specific medium to cultivate and establish intestinal organoids of livestock. Species-specific medium is applied differently between species for the cultivation of intestinal organoids, and its modification is important for the maintenance of specific cell types or genes from the cellular diversity of the intestinal epithelium. In this study, we introduce the histological development of a 3D culture system and a species-specific medium for the cultivation of intestinal organoids in livestock. Finally, we discuss the importance and future perspectives of intestinal organoids in the fields of agriculture and biotechnology for various purposes.
This study investigated various levels of sodium nitrite and probiotics (SNPro) combination as an alternative to zinc oxide on the growth performance, immune response, intestinal microflora, and morphology of weaned pigs. One hundred and ninety-two weaned pigs (Landrace×Yorkshire×Duroc) with an average body weight of 6.51±0.15 kg were randomly assigned to four treatments(n=6) on the basis of their initial body weight. Experimental period was divided into phase 1 and 2 (each 14 days). The dietary treatments were: 1) Basal diet (control), 2) SNPro1 (control+0.01% SNPro), 3) SNPro2 (control+0.02% SNPro), 4) SNPro3 (control+0.03% SNPro). The average daily gain when SNPro was added to the diet was 288, 309, 319, 324 g in phase 1, 355, 387, 410, 407 g in phase 2 and 321, 348, 364, 366 g in the overall. The concentration of interleukin-8 and interleukin-10 in serum when SNPro was added to the diet were 15, 13.5, 13, 12.8 ng/ml and 165, 162, 155, 145 ng/ml (p<0.05) but toll-like receptor 4 and immunoglobulin G levels in serum were no significantly different. The colonization of Escherichia coli in the ileum and Salmonella spp. in the caecum were significantly decreased as SNPro level increased (p<0.05). However, the population of Lactobacillus spp. did not differ among the groups. Although villus height and villus height to crypt depth ratio were not significantly affected by the treatments, crypt depth in the jejunum was 599, 586, 615, 599 ㎛ as SNPro level increased (p<0.05). In conclusion, SNPro had beneficial effects on growth performance, immune response, intestinal microflora and morphology weaned pigs. Therefore, SNPro not only can be considered as an alternative for the pharmacological level of zinc oxide in weaning pigs but also ideal dietary SNPro level was 0.02%.
This study was conducted to determine the effects of dietary protein level and supplementation of protease on growth performance, nutrient digestibility, gut microflora, intestinal morphology and fecal noxious gas emission in weanling pigs. A total of 240 weaned pigs (Landrace×Yorkshire×Duroc, 5.82±0.3 kg) were used during 4 weeks in 2 phases (days 0-14, phase 1; and days 15-28, phase 2) feeding program based on age and initial body weight. Pigs were allocated to 2×2 factorial arrangement, including 2 protein levels (HP, high protein; LP, low protein) and 2 protease levels (with or without protease). The average daily gain in the LP treatment (357 g/d) was increased rather than the HP treatment (339 g/d). A greater avarage daily gain was observed in dietary suppiemented protease treatment (358 vs 339 g/d). Average feed intake was greater in the LP treatment (544 g/d) rather than the HP treatment (530 g/d). A greater average daily feed intake was observed in dietary supplemented protease treatment (552 vs 523 g/d). Dry matter and crude protein digestibility were increased in dietary supplemented protease treatment (82.62% and 76.08%, respectively) rather than non-supplemented treatment (81.74% and 75.13%, respectively). Ileal Lactobacillus spp. count increased in dietary supplemented protease treatment (7.42 vs 7.32 log10CFU/g). Emission of H2S was decreased in the LP treatment (4.41 ppm) rather than HP treatment (4.78 ppm). Emission of NH3 was decreased in dietary supplemented protease treatment (10.43 ppm vs 11.76 ppm). In conclusion, the decrease of dietary protein level and supplementation of protease had beneficial effects on growth performance, nutrient digestibility, gut microflora, and noxious gas emission in weanling pigs.
Celecoxib, a cyclooxygenase (COX)-2 selective inhibitor, was approved as a non-steroidal anti-inflammatory drug (NSAID), and this therapeutic application has been expanded to several other diseases, including colon cancer. Notably, a treatment strategy combining the use of celecoxib and radiation therapy has been employed for improving the control of local cancers. In this study, we examined the effect of celecoxib on irradiation-induced intestinal damage. The twenty four mice (BALB/c) were divided into four groups; 1) sham-irradiated control group, 2) celecoxib-treated group, 3) irradiated group, and 4) celecoxib-treated irradiation group. Mice were orally administered celecoxib at a dose of 25 mg/kg in a 0.1 mL volume, daily for 4 days after irradiation exposure (10 Gy). Then, histological examinations of the jejunal villous height, crypt survival, and crypt size were performed. The expression of COX-2 after administration of celecoxib in irradiated mice was examined by employing immunohistochemistry, Western blotting, and qPCR analysis. The jejunal villi height and the crypt survival were reduced in the irradiation group compared with the sham-irradiated group. Celecoxib treatment in irradiation mice even more decreased those indicators. Crypt size was increased in the radiation group compared to the sham-irradiated control group, whereas the size was decreased in the celecoxibtreated irradiation group compared with the group exposed to the radiation injury. COX-2 expression was detected in the crypt of the small intestine, and COX-2 expression was increased in the crypt lesion following radiation exposure. However, COX-2 expression was reduced in the celecoxib-treated irradiation group. Therefore, in the present study, we confirmed that celecoxib treatment after irradiation aggravated the irradiation-induced intestinal damage. These results suggest that a caution need to be administered when celecoxib treatment is performed in combination with radiation therapy for cancer treatment.
이 연구는 복분자에서 분리된 Lactobacillus plantarum stain (GBL16 및 17)을 포함한 probiotic에 대해 수행되었으며, 장내 미생물 변화를 확인하였다. Balb/c 마우스모델에서 GBL 16, 17을 포함한 시제품인 PMC와 시판중인 유산균 과자 BFY를 8주간 경구 투여 하였다. PMC의 투여는 장내 미생물총의 균형을 향상시키고, 장내개선에 긍정적인 영향을 보였다. 맹장을 이용한 NGS 분석을 통해 확인한 결과, 스트레스 후 증가하는 양상을 보이는 균인 Lachnospiraceae균이 감소하는 경향을 보였다. Alistipes균의 증가는 염증성 장 질환에 효과가 있을 것으로 판단되나, 추후 더 많은 연구가 필요할 것으로 보인다.