immune; intestinal bacteria; mannan oligosaccharide; piglet; sow. J Anim Sci. gut microbiota; konjac mannan-oligosaccharides (MOS); metformin; type 2 diabetes. The structure of action of the biologically active MOS species should be determined and mechanism of action further investigated. 3B). (Fig. Here, oligosaccharides take on a new role—that of a prebiotic. The effects of M5 over cellular tight junction was distinguished from the cellular proliferation by calcium depletion experiment where calcium ions were removed from the culture medium once the growth of the cell reached 85% confluency, replacing DMEM with SMEM. 2019 Jul 24;11(8):1705. doi: 10.3390/nu11081705. Determination of biological active MOS species was also investigated. The reaction contained SDS or EDTA decreases the activity of RMase24 to 25% and 44%, respectively, while Triton X-100 had no effects on RMase24 activity (Fig. 1B). Although the identification and the mechanism of actions of MOS remained unclear, existing evidence strongly suggests that MOS may exert biological effects on the intestine regarding tissue compact and increased in structure and activity. MOS produced may also require additional purification process to remove the acid and chemical contaminants. In contrast, digestion of G-GalMan did not change the overall structure observed by SEM much (Figs. A total of 60 multiparous sows (4 ± 1 parity; Landrace × Yorkshire) were fed either control diet (sCON, n = 30) or a diet containing 400 mg kg-1 MOS (sMOS, n = 30) from day 86 of gestation until weaning (day 20 of postpartum). 4A) and their molecular mass confirmed by mass spectrometry. "Following" is like subscribing to any updates related to a publication. Hamilton MK, Ronveaux CC, Rust BM, Newman JW, Hawley M, Barile D, Mills DA, Raybould HE. National Center for Biotechnology Information, Unable to load your collection due to an error, Unable to load your delegates due to an error. The activity was measured using DNS assay using glucose as a standard (Miller, 1959). A finely ground copra meal powder was then subjected to solvent extraction to remove the remaining oil with 200 milliliters of n-hexane, and with sonication for 10 min. The percentages yield of total MOS produced from S-GalMan and G-GalMan were 8.25% and 5.09% (dry basis), respectively. National Center for Biotechnology Information, Unable to load your collection due to an error, Unable to load your delegates due to an error. All the substrates were subjected to critical point dryer (CPD) and were coated with gold particles prior to the scanning electron microscopy (SEM-EDS) (Jeol JSM-6400 scanning electron microscope; Jeol Ltd., Tokyo, Japan). The results showed that the enzyme/substrate ratio used in the digestion affected the pattern of MOSs produced (Fig. The authors declare there are no competing interests. Ammonium sulfate was added to give the final concentration of 1 then it was applied onto phenyl toyopearl column. Thin layer chromatography analysis of digested products showed a spectrum of MOS with size ranging from DP2 to DP6 and MOS with a size larger than DP6, comparing to standard MOS ladder. Thus, the biologically active MOS species were not identified. Xu S, Shi J, Shi X, Dong Y, Wu X, Li Z, Fang Z, Lin Y, Che L, Li J, Feng B, Wang J, Wu D, Shen Y. J Anim Sci. Epub 2019 Apr 20. The percentage yield was calculated to compare the efficiency between each type of substrate pretreatment. Nopvichai C, Charoenwongpaiboon T, Luengluepunya N, Ito K, Muanprasat C, Pichyangkura R. (A) and (B) reveal the optimum pH and temperature of purified RMase24, respectively. Effects of dietary fiber sources during late gestation and lactation on sow performance, milk quality, and intestinal health in piglets1. MOS is another controversial feed additive that has drawn much attention in the last decade. HHS Fouhse JM, Dawson K, Graugnard D, Dyck M, Willing BP. These types of mannan consist of (1-4)-beta-D-mannose repeating units with (1-6)-alpha-D-galactose units attached to the mannose backbone (Ganter et al., 1995; McCleary, Matheson & Small, 1976; Sittikijyothin, Torres & Gonçalves, 2005).  |  Pascale A, Marchesi N, Govoni S, Coppola A, Gazzaruso C. Curr Opin Pharmacol. The fractions with enzymatic activity were pooled together. Get the latest public health information from CDC: https://www.coronavirus.gov. The polymerase chain reaction was performed using 5′-GGGGAGTTGCATATGTTTAAGA-3′and 5′-GCGGAACGTCTGATTAGAGC-3′ as a forward primer and reverse primer, respectively. On day 7 of age, offspring (Duroc × Landrace Yorkshire) were assigned within sow treatments and fed control diet (pCON) or diet containing 800 mg kg-1 MOS (pMOS) for 28 d (end at 35 d of age), resulting in four piglet diet groups (n = 15 litters per diet group): sCON-pCON, sCON-pMOS, sMOS-pCON, and sMOS-pMOS. FASEB J. eCollection 2020. Clipboard, Search History, and several other advanced features are temporarily unavailable. Un oligosaccharide est un glucide formé de 2 à 20 monosaccharides environ. Transwell (Corning, Corning, NY, USA) culture plates and measured the TEER daily. The interaction effects between sows and piglet diets on IL-4 (P = 0.02), IL-10 (P < 0.01), and IFN-γ (P = 0.08) were observed. TEER at different time intervals were measured to monitor the reassembling of the tight junction for 12 h. Statistical analysis was performed using GraphPad prism7 (GraphPad Software Inc. La Jolla, CA, USA). This finding indicated that M5 was playing the vital role in increasing the tight junction of epithelial tissue. These results may result from the higher exposed surface area arose from the additional sonication step that is accessible to enzymatic hydrolysis. The following information was supplied regarding the deposition of DNA sequences: The DNA sequencing data is available at GenBank: KY951415. Dietary supplementation of weaned piglets with a yeast-derived mannan-rich fraction modulates cecal microbial profiles, jejunal morphology and gene expression. The measurement protocol was described in detail in our previous study (Muanprasat et al., 2015). Epub 2016 Dec 15. Results found that piglet diet MOS increased or tend to increase Lactobacillus amount in the ileum digesta (P < 0.01) and jejunum digesta (P = 0.07), respectively; while tend to decrease Escherichia coli amount in jejunum digesta (P =0.06) and cecum digesta (P = 0.08). All rights reserved. USA.gov. NIH Let’s take a closer look at what MOS does and why it’s so important for your horse’s health. By Ioannis Mavromichalis | November 19, 2014. The recombinant enzyme was named RMase24. For pH stability analysis, the residual enzymatic activity was measured by preincubating the enzyme at 4 °C for 24 h at various pH described above. The mannanase gene was cloned from Bacillus subtilis cAE24, then expressed in BL21. Determination of appropriate RMase24 units required for optimum MOS production was then performed with S-GalMan to determine the optimal enzyme/substrate ratio. You can add specific subject areas through your profile settings. 2019 Sep 3;97(9):3958-3971. doi: 10.1093/jas/skz242. The suspension was filtered with a cellulose filter paper (Whatman no.1, Sigma Aldrich, USA.). Role of gut microbiota in type 2 diabetes pathophysiology. The interaction effects between sow and piglet diets were found on the mRNA levels of NF- κB p65 (P = 0.03) and IL-8 (P = 0.02) in piglet jejunum. Each purified MOS ranging from DP4 to DP7 (M4-M7) was filter-sterilized through 0.2 micrometer Whatman uniflo syringe filter (GE Healthcare, Chicago, IL, USA) before being made up to 10 micromolar solutions with Dulbecco’s Modified Eagle’s Medium (DMEM, Corning life science, USA). 2C–2D). The activity unit was defined as the amount of enzyme that is required to release 1 micromole of reducing sugar from 0.5%w/v glucomannan substrate per minute. The production of MOS from galactomannan can be performed either by acid or enzymatic hydrolysis. Mannan oligosaccharides (MOS) and β-glucans have been reported to beneficially influence broiler performance and health. Typos, corrections needed, missing information, abuse, etc. 2016 Dec;116(11):1869-1877. doi: 10.1017/S0007114516004086. Bacillus subtilis is also used in a traditional fermented soybean food (e.g., Natto) in Japan (Lefevre et al., 2017; Wang & Fung, 1996). T84 cells were cultured in DMEM medium until the population of the cells reached 85% confluency on transwell plate, then DMEM medium was substituted with SMEM to remove Ca2+ ions. The arrow in Figs. However, different purifications levels have been employed to investigate the efficacy of those oligosaccharides, and thus conflicts have been noticed with respect to their influences. 2019 Nov;33(11):11655-11667. doi: 10.1096/fj.201802802RRR. Les oligosaccharides sont des molécules formées par la liaison de monosaccharides cycliques, linéaires ou ramifiés par des liaisons de type glycosidique, une liaison covalente s'établissant entre des groupes alcooliques de deux monosaccharides, avec libération d'une molécule d'eau. 2A–2B). Once the electrical resistant of the cells became steady, each prepared MOS was treated to the cells. Cao TTB, Wu KC, Hsu JL, Chang CS, Chou C, Lin CY, Liao YM, Lin PC, Yang LY, Lin HW. All the deoiled substrates, including G-GalMan, S-Galman, and digested products were prepared as previously described. Hereafter, further investigation is required to identify the structure and mechanism of action of M5. The effects of metal ions also were investigated. The gene sequence of RMase24 was 1,090 bp in size, encoding 362 amino acid residues (Data S2). Navaporn Luengluepunya performed the experiments, bacillus subtilis cAE24 screening. It was identified by 16s rRNA sequence comparison, which showed a 99% sequence identity with Bacillus subtillis strain W_6 (accession number JX462604.1). MOS DP4 (M4) to DP7 (M7) was successfully purified through Biogel-P2 column (Fig. The ions that show the highest enhancing effects to RMase24 were cobalt and zinc ions, which increased its activity to about 136 and 137 percent, respectively. Because the first line clinical medicine of metformin has several intestinal drawbacks, combination usage of metformin with a prebiotic of konjac mannan-oligosaccharides (MOS) was conceived and implemented aiming to investigate whether there were some intestinal synergetic effects and how MOS would function. Epub 2019 Aug 15. Our promise Several reports revealed that MOS supplement could improve growth performance and body weight in various animals (Ai et al., 2011; Dimitroglou et al., 2010; Genc et al., 2007; Mansour et al., 2012; Staykov et al., 2007). Scanning electron microscope imaging showed that pretreatment using sonication could disrupt the surface of copra meal better than grinding alone, which can improve the production of MOS. Gurung M, Li Z, You H, Rodrigues R, Jump DB, Morgun A, Shulzhenko N. EBioMedicine. At higher magnification, different characters of fragment surface and edges from each type of substrate was observed (Figs. Please enable it to take advantage of the complete set of features! Sequemce analysis result of cAE24 and 16s rRNA sequence comparison are available in (Data S1 and Fig. We have successfully isolated bacterial strain with high mannanase activity. Sonication pretreatment gave smaller average fragment size when compared with the untreated substrate (Figs. RMASE24 represents the protein sequence of RMase24. At high enzyme over substrate ratio, the availability free enzyme in the solution may cause further hydrolysis of some MOS products that remains a good substrate for the enzyme. A comparison between a digestion of S-GalMan (A) and G-GalMan (B) with 10 units per 1 gram of substrate. A total of 60 multiparous sows (4 ± 1 parity; Landrace × Yorkshire) … Effects of supplementing processed straw during late gestation on sow physiology, lactation feed intake, and offspring body weight and carcass quality1.