Supplementing Glycerol to Inoculum Induces Changes in pH, SCFA Profiles and Microbiota Composition in In-Vitro Batch Fermentation

Glycerol was generally added to the inoculum as a cryoprotectant. However, it was also a suitable substrate for microbial fermentation, which may produce more SCFAs, thereby decreased pH of the fermentation broth. This study investigated the effect of supplementing glycerol to inoculum on in vitro fermentation and whether an enhanced buffer capacity of medium could maintain the pH stability during in vitro batch fermentation, subsequently improving the accuracy of short chain fatty acids (SCFAs) determination, especially propionate. Two ileal digesta were fermented by pig fecal inoculum with or without glycerol (served as anti-frozen inoculum or frozen inoculum) in standard buffer or enhanced buffer solution (served as normal or modified medium). Along with the fermentation, adding glycerol decreased the pH of fermentation broth (p < 0.05). However, modified medium could alleviate the pH decrement compared with normal medium (p < 0.05). The concentration of total propionic acid production was much higher than that of other SCFAs in anti-frozen inoculum fermentation at 24 and 36 h, thereby increasing the variation (SD) of net production of propionate. The α-diversity analysis showed that adding glycerol decreased Chao1 and Shannon index under normal medium fermentation (p < 0.05) compared to modified medium (p < 0.05) along with fermentation. PCoA showed that all groups were clustered differently (p < 0.01). Adding glycerol improved the relative abundances of Firmicutes, Anaerovibrio, unclassified_f_Selenomonadaceae, and decreased the relative abundance of Proteobacteria (p < 0.05). The relative abundances of Firmicutes, such as Lactobacillus, Blautia and Eubacterium_Ruminantium_group in modified medium with frozen inoculum fermentation were higher than (p < 0.05) those in normal medium at 36 h of incubation. These results showed that adding glycerol in inoculum changed the fermentation patterns, regardless of substrate and medium, and suggested fermentation using frozen inoculum with modified medium could maintain stability of pH, improve the accuracy of SCFA determination, as well as maintain a balanced microbial community.

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Cross-Talk Between Butyric Acid and Gut Microbiota in Ulcerative Colitis Following Fecal Microbiota Transplantation

Frontiers in Microbiology ◽

10.3389/fmicb.2021.658292 ◽

2021 ◽

Vol 12 ◽

Author(s):

Hao-Ming Xu ◽

Hong-Li Huang ◽

Jing Xu ◽

Jie He ◽

Chong Zhao ◽

...

Keyword(s):

Ulcerative Colitis ◽

Gut Microbiota ◽

Butyric Acid ◽

Fecal Microbiota Transplantation ◽

Short Chain Fatty Acids ◽

Fecal Microbiota ◽

16S Sequencing ◽

Α Diversity ◽

The Impact

Fecal microbiota transplantation (FMT) can inhibit the progression of ulcerative colitis (UC). However, how FMT modulates the gut microbiota and which biomarker is valuable for evaluating the efficacy of FMT have not been clarified. This study aimed to determine the changes in the gut microbiota and their relationship with butyric acid following FMT for UC. Fecal microbiota (FM) was isolated from healthy individuals or mice and transplanted into 12 UC patients or colitis mice induced by dextran sulfate sodium (DSS). Their clinical colitis severities were monitored. Their gut microbiota were analyzed by 16S sequencing and bioinformatics. The levels of fecal short-chain fatty acids (SCFAs) from five UC patients with recurrent symptoms after FMT and individual mice were quantified by liquid chromatography–mass spectrometry (LC–MS). The impact of butyric acid on the abundance and diversity of the gut microbiota was tested in vitro. The effect of the combination of butyric acid-producing bacterium and FMT on the clinical responses of 45 UC patients was retrospectively analyzed. Compared with that in the controls, the FMT significantly increased the abundance of butyric acid-producing bacteria and fecal butyric acid levels in UC patients. The FMT significantly increased the α-diversity, changed gut microbial structure, and elevated fecal butyric acid levels in colitis mice. Anaerobic culture with butyrate significantly increased the α-diversity of the gut microbiota from colitis mice and changed their structure. FMT combination with Clostridium butyricum-containing probiotics significantly prolonged the UC remission in the clinic. Therefore, fecal butyric acid level may be a biomarker for evaluating the efficacy of FMT for UC, and addition of butyrate-producing bacteria may prolong the therapeutic effect of FMT on UC by changing the gut microbiota.

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Impact of Grain Sorghum Polyphenols on Microbiota of Normal Weight and Overweight/Obese Subjects during In Vitro Fecal Fermentation

Nutrients ◽

10.3390/nu11020217 ◽

2019 ◽

Vol 11 (2) ◽

pp. 217 ◽

Cited By ~ 7

Author(s):

Danielle Ashley ◽

Daya Marasini ◽

Cindi Brownmiller ◽

Jung Lee ◽

Franck Carbonero ◽

...

Keyword(s):

Gut Microbiota ◽

Batch Fermentation ◽

Short Chain Fatty Acids ◽

Negative Control ◽

Normal Weight ◽

Weight Class ◽

Gut Health ◽

Sorghum Bran ◽

Before And After

The human gut microbiota is considered as a crucial mediator between diet and gut homeostasis and body weight. The unique polyphenolic profile of sorghum bran may promote gastrointestinal health by modulating the microbiota. This study evaluated gut microbiota and modulation of short-chain fatty acids (SCFA) by sorghum bran polyphenols in in vitro batch fermentation derived from normal weight (NW, n = 11) and overweight/obese (OO, n = 11) subjects’ fecal samples. Six separate treatments were applied on each batch fermentation: negative control (NC), fructooligosaccharides (FOS), black sorghum bran extract (BSE), sumac sorghum bran extract (SSE), FOS + BSE, or FOS + SSE; and samples were collected before and after 24 h. No significant differences in total and individual SCFA production were observed between NW and OO subjects. Differential responses to treatment according to weight class were observed in both phyla and genera. Sorghum bran polyphenols worked with FOS to enhance Bifidobacterium and Lactobacillus, and independently stimulated Roseburia and Prevotella (p < 0.05). Our results indicate that sorghum bran polyphenols have differential effects on gut health and may positively impact gut ecology, with responses varying depending on weight class.

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Identification of Enterotype and Its Effects on Intestinal Butyrate Production in Pigs

Animals ◽

10.3390/ani11030730 ◽

2021 ◽

Vol 11 (3) ◽

pp. 730

Author(s):

E Xu ◽

Hua Yang ◽

Minmin Ren ◽

Yuanxia Wang ◽

Mingfei Xiao ◽

...

Keyword(s):

High Throughput Sequencing ◽

Bacterial Community Composition ◽

Short Chain Fatty Acids ◽

Diversity Indices ◽

Sensu Stricto ◽

Shannon Index ◽

Nutrient Metabolism ◽

Fecal Samples ◽

Coa Transferase ◽

Α Diversity

Gut microbiota is thought to play a crucial role in nutrient digestion for pigs, especially in processing indigestible polysaccharides in the diets to produce short-chain fatty acids (SCFAs). However, the link between microbiota community structure and phenotypic performances are poorly understood. In the present study, the fecal samples of 105 Jinhua pigs at 105 days of age were clustered into three enterotypes (ETs, ET1, ET2, and ET3) that are subpopulations of distinct bacterial community composition by using 16S rRNA high throughput sequencing. The α-diversity indices (the OTU number and Shannon index) were significantly different among the ETs (p < 0.001). At the genus level, the ET1 group was over-represented by Lactobacillus (17.49%) and Clostridium sensu stricto 1 (11.78%), the ET2 group was over-represented by Clostridium sensu stricto 1 (17.49%) and Bifidobacterium (11.78%), and the ET3 group was over-represented by Bacteroides (18.17%). Significant differences in the fecal contents of butyrate were observed among ETs, with the highest level detected in ET3 and the lowest in ET2 (p < 0.05). Consistently, more copies of the terminal genes for butyrate synthesis, butyrate kinase (Buk) and butyryl coenzyme A (CoA): acetate CoA transferase (But) were detected by qPCR in the fecal samples of the ET3 group as compared to other two groups (p < 0.05). In addition, of the two genes, But was demonstrated to be more relevant to the butyrate content (R = 0.7464) than Buk (R = 0.4905) by correlation analysis. In addition, based on the taxonomic analysis, we found that Faecalibacterium was the most relevant butyrate-producing genera with fecal butyrate contents in Jinhua pigs, followed by Butyricicoccus, Eubacterium, Butyricimonas, Blautia, and Anaerostipes, all of which showed significantly higher richness in ET3 than as compared to ET1 and ET2 (p < 0.05). Collectively, this work presents a first overview of the enterotypes clustering in Jinhua pigs and will help to unravel the functional implications of ETs for the pig’s phenotypic performance and nutrient metabolism.

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Alterations in the Blood Parameters and Fecal Microbiota and Metabolites during Pregnant and Lactating Stages in Bama Mini Pigs as a Model

Mediators of Inflammation ◽

10.1155/2020/8829072 ◽

2020 ◽

Vol 2020 ◽

pp. 1-13

Author(s):

Cui Ma ◽

QianKun Gao ◽

WangHong Zhang ◽

Md. Abul Kalam Azad ◽

XiangFeng Kong

Keyword(s):

Relative Abundance ◽

Short Chain Fatty Acids ◽

Fecal Microbiota ◽

Reproductive Hormones ◽

Plasma Prolactin ◽

Microbiota Composition ◽

Α Diversity ◽

Lactating Sows ◽

Pregnancy And Lactation ◽

Relative Abundances

This study was conducted to analyze plasma reproductive hormone and biochemical parameter changes, as well as fecal microbiota composition and metabolites in sows, at different pregnancy and lactation stages, using Bama mini pig as an experimental animal model. We found that plasma prolactin (PRL), progesterone, follicle-stimulating hormone (FSH), and estrogen levels decreased from day 45 to day 105 of pregnancy. Plasma total protein and albumin levels were lower in pregnant sows, while glucose, urea nitrogen, total cholesterol, and high-density lipoprotein-cholesterol, as well as fecal acetate, butyrate, valerate, total short-chain fatty acids, skatole, and tyramine levels, were higher in lactating sows. Interestingly, the lactating sows showed lower α-diversity and Spirochaetes and Verrucomicrobia relative abundances, while pregnant sows showed a higher Proteobacteria relative abundance. Notably, the Akkermansia relative abundance was highest on day 7 of lactation. Spearman analysis showed a positive correlation between plasma triglyceride and cholinesterase levels and Akkermansia and Streptococcus relative abundances. Moreover, Oscillospira and Desulfovibrio relative abundances were also positively correlated with plasma FSH, LH, and E2 levels, as well as PRL and LH with Bacteroides. Collectively, plasma reproductive hormones, biochemical parameters, and fecal microbiota composition and metabolite levels could alter along with pregnancy and lactation, which might contribute to the growth and development demands of fetuses and newborns.

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Short-Chain Fatty Acids Prevent Diabetic Nephropathy In Vivo and In Vitro

Diabetes ◽

10.2337/db18-92-or ◽

2018 ◽

Vol 67 (Supplement 1) ◽

pp. 92-OR ◽

Cited By ~ 1

Author(s):

WEI HUANG ◽

YONG XU ◽

YOUHUA XU ◽

LUPING ZHOU ◽

CHENLIN GAO

Keyword(s):

Fatty Acids ◽

Diabetic Nephropathy ◽

Short Chain Fatty Acids ◽

Short Chain ◽

Chain Fatty Acids

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Improving Topical Skin Delivery of Monocrotaline Via Liposome Gel-based Nanosystems

Current Drug Delivery ◽

10.2174/1567201816666191029125300 ◽

2019 ◽

Vol 16 (10) ◽

pp. 940-950 ◽

Cited By ~ 1

Author(s):

Jiandong Yu ◽

Zhi Chen ◽

Yan-zhi Yin ◽

Chaoyuan Tang ◽

Enying Hu ◽

...

Keyword(s):

Gradient Method ◽

Encapsulation Efficiency ◽

Ph Value ◽

Topical Administration ◽

Ph Gradient ◽

Stability Factor ◽

Buffer Solution ◽

Skin Delivery ◽

Skin Retention

Background: In this study, a liposomal gel based on a pH-gradient method was used to increase the skin-layer retention of monocrotaline (MCT) for topical administration. Methods: Using the Box-Behnken design, different formulations were designed to form liposome suspensions with optimal encapsulation efficiency (EE%) and stability factor (KE). In order to keep MCT in liposomes and accumulate in skin slowly and selectively, MCT liposome suspensions were engineered into gels. Results: A pH-gradient method was used to prepare liposome suspensions. The optimal formulation of liposome suspensions (encapsulation efficiency: 83.10 ± 0.21%) was as follows: MCT 12 mg, soybean phosphatidyl choline (sbPC) 200 mg, cholesterol (CH) 41 mg, vitamin E (VE) 5 mg, and citric acid buffer solution (CBS) 4.0 10 mL (pH 7.0). The final formulation of liposomal gels consisted of 32 mL liposome suspensions, 4.76 mL deionized water, 0.40 g Carbopol-940, 1.6 g glycerol, 0.04 g methylparaben, and a suitable amount of triethanolamine for pH value adjustment. The results of in vitro drug release showed that MCT in liposomal gels could be released in 12 h constantly in physiological saline as a Ritger-Peppas model. Compared with plain MCT in gel form, liposomal MCT in gel had higher skin retention in vitro. Conclusion: In this study, liposomal gels were formed for greater skin retention of MCT. It is potentially beneficial for reducing toxicities of MCT by topical administration with liposomal gel.

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Chemical composition and in vitro digestibility of some selected under-utilized tropical seeds as protein sources in ruminants’ diet

Bulletin of the National Research Centre ◽

10.1186/s42269-020-00430-9 ◽

2020 ◽

Vol 44 (1) ◽

Author(s):

Oluwatosin Bode Omotoso ◽

Mary Oluwafunmilayo Adeduntan ◽

Adebowale Noah Fajemisin

Keyword(s):

Short Chain Fatty Acids ◽

Nutritional Composition ◽

Metabolizable Energy ◽

In Vitro Digestibility ◽

Protein Sources ◽

Dry Matter Digestibility ◽

Supplementary Feed ◽

Monodora Myristica

Abstract Background The study highlighted the potential of three common and under-utilized tropical leguminous seeds (Tomentosa nilotica, Dioclea reflexa and Monodora myristica) to be used as supplementary feed to ruminant livestock. These seeds (their plants inclusive) are valuable sources of food and medicine for the prevention of illness and maintenance of human health. The medicinal properties of these seeds include antimicrobial, anti-inflammatory, anti-oxidant and immuno-stimulant. Trypsin inhibitors, which are common anti-nutritional factors in legumes and for monogastric animals, do not exert adverse effects in ruminants because they are degraded in the rumen. Hence, the crux of this study is to examine the effect of processing methods on the nutritional composition (proximate, fibre fractions, minerals, anti-nutrients) and in vitro digestibility of Tomentosa nilotica, Dioclea reflexa and Monodora myristica seeds and their suitability as feedstuff (protein sources) in small ruminant feed, particularly during off-season. Results From the results, raw Tomentosa nilotica and Monodora myristica have the highest crude protein (30.35% CP) and fat (22.40% EE), respectively. It is noteworthy that roasting best improve the mineral and significantly reduce the anti-nutrients observed in this study better compared to boiling and soaking methods. The highest organic matter digestibility, short-chain fatty acids, metabolizable energy and in vitro dry matter digestibility values were obtained in Dioclea reflexa compared to other test seeds. Roasting best improved the nutritive values, while Dioclea reflexa seed was rated highest for all the nutritional attributes and in vitro digestibility. Conclusions Dioclea reflexa could be incorporated in ruminants’ diet as protein source, particularly during the off-season, for improved ruminant production in Nigeria. However, in vivo study is therefore recommended to validate this report.

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Feeding Value Assessment of Substituting Cassava (Manihot esculenta) Residue for Concentrate of Dairy Cows Using an In Vitro Gas Test

Animals ◽

10.3390/ani11020307 ◽

2021 ◽

Vol 11 (2) ◽

pp. 307

Author(s):

Yuhui Zheng ◽

Yanyan Zhao ◽

Shenglin Xue ◽

Wei Wang ◽

Yajing Wang ◽

...

Keyword(s):

Manihot Esculenta ◽

Rumen Fluid ◽

Gas Production ◽

The Other ◽

Holstein Cows ◽

Value Assessment ◽

Buffer Solution ◽

Feeding Value ◽

Cassava Residue

The feeding value of replacing concentrate with cassava (Manihot esculenta) residue in the feed of Holstein cows was confirmed using an in vitro gas test. The treatments consisted of 0% (control, CON), 5%, 10%, 15%, 20%, 25%, and 30% inclusion of cassava residue in fermentation culture medium composed of buffer solution (50 mL) and filtrated rumen fluid (25 mL). The parameters analyzed included the kinetics of gas production and fermentation indexes. Forty-eight hours later, there were no significant differences on in vitro dry matter disappearance (IVDMD), pH, and microbial crude protein (MCP) content among treatments (p > 0.05). However, the “cumulative gas production at 48 h” (GP48), the “asymptotic gas production” (A), and the “maximum gas production rate” (RmaxG) all increased linearly or quadratically (p < 0.01). The GP48 was significantly higher in the 25% treatment compared to the other treatments, except for the 30% (p < 0.01). The A was significantly larger in the 25% treatment compared to the other treatments, except for the 20% and 30% (p < 0.01). The RmaxG was distinctly larger in the 25% treatment compared to other treatments (p < 0.01); moreover, the “time at which RmaxG is reached” (TRmaxG) and the “time at which the maximum rate of substrate degradation is reached” (TRmaxS) were significantly higher in the 25% treatment than the CON, 20%, and 30% treatments (p < 0.01). Additionally, the content of ammonia-N (NH3-N) in all treatments showed linearly and quadratically decreases (p < 0.01), whereas total volatile fatty acid (VFA), iso-butyrate, butyrate, and iso-valerate contents changed quadratically (p = 0.02, p = 0.05, p = 0.01, and p = 0.02, respectively); all of these values peaked in the 25% treatment. In summary, the 25% treatment was associated with more in vitro gas and VFA production, indicating that this cassava residue inclusion level may be used to replace concentrate in the feed of Holstein cows. However, these results need to be verified in vivo.

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Dietary Fiber Modulates the Fermentation Patterns of Cyanidin-3-O-Glucoside in a Fiber-Type Dependent Manner

Foods ◽

10.3390/foods10061386 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1386

Author(s):

Zixin Yang ◽

Ting Huang ◽

Ping Li ◽

Jian Ai ◽

Jiaxin Liu ◽

...

Keyword(s):

Dietary Fiber ◽

Fiber Type ◽

Short Chain Fatty Acids ◽

Dependent Manner ◽

Dietary Fibers ◽

Cell Wall Polysaccharides ◽

Total Carbohydrate ◽

In Vitro Fermentation ◽

Gas Volume

The interactions between cell-wall polysaccharides and polyphenols in the gastrointestinal tract have attracted extensive attention. We hypothesized that dietary fiber modulates the fermentation patterns of cyanidin-3-O-glucoside (C3G) in a fiber-type-dependent manner. In the present study, the effects of four dietary fibers (fructose-oligosaccharides, pectin, β-glucan and arabinoxylan) on the modulation of C3G fermentation patterns were investigated through in vitro fermentation inoculated with human feces. The changes in gas volume, pH, total carbohydrate content, metabolites of C3G, antioxidant activity, and microbial community distribution during in vitro fermentation were analyzed. After 24 h of fermentation, the gas volume and total carbohydrate contents of the four dietary-fiber-supplemented groups respectively increased and decreased to varying degrees. The results showed that the C3G metabolites after in vitro fermentation mainly included cyanidin, protocatechuic acid, 2,4,6-trihydroxybenzoic acid, and 2,4,6-trihydroxybenzaldehyde. Supplementation of dietary fibers changed the proportions of C3G metabolites depending on the structures. Dietary fibers increased the production of short-chain fatty acids and the relative abundance of gut microbiota Bifidobacterium and Lactobacillus, thus potentially maintaining colonic health to a certain extent. In conclusion, the used dietary fibers modulate the fermentation patterns of C3G in a fiber-type-dependent manner.

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Impact of 2′-Fucosyllactose on Gut Microbiota Composition in Adults with Chronic Gastrointestinal Conditions: Batch Culture Fermentation Model and Pilot Clinical Trial Findings

Nutrients ◽

10.3390/nu13030938 ◽

2021 ◽

Vol 13 (3) ◽

pp. 938

Author(s):

Jennifer Joan Ryan ◽

Andrea Monteagudo-Mera ◽

Nikhat Contractor ◽

Glenn R. Gibson

Keyword(s):

Ulcerative Colitis ◽

Batch Culture ◽

Pilot Trial ◽

Gastrointestinal Symptoms ◽

Short Chain Fatty Acids ◽

Open Label ◽

Gastrointestinal Conditions ◽

Fermentation Model ◽

Quality Of Life Index

Intestinal dysbiosis has been described in patients with certain gastrointestinal conditions including irritable bowel syndrome (IBS) and ulcerative colitis. 2′-fucosyllactose (2′-FL), a prebiotic human milk oligosaccharide, is considered bifidogenic and butyrogenic. To assess prebiotic effects of 2′-FL, alone or in combination with probiotic strains (potential synbiotics), in vitro experiments were conducted on stool from healthy, IBS, and ulcerative colitis adult donors. In anaerobic batch culture fermenters, Bifidobacterium and Eubacterium rectale-Clostridium coccoides counts, and short-chain fatty acids (SCFAs) including butyrate increased during fermentation with 2′-FL and some of the 2′-FL/probiotic combinations. In a subsequent open-label pilot trial, the effect of a 2′-FL-containing nutritional formula was evaluated in twelve adults with IBS or ulcerative colitis. Gastrointestinal Quality of Life Index (GIQLI) total and gastrointestinal symptoms domain scores, stool counts of Bifidobacterium and Faecalibacterium prausnitzii, and stool SCFAs including butyrate, increased after six weeks of intervention. Consistent with documented effects of 2′-FL, the batch culture fermentation experiments demonstrated bifidogenic and butyrogenic effects of 2′-FL during fermentation with human stool samples. Consumption of the 2′-FL-containing nutritional formula by adults with IBS or ulcerative colitis was associated with improvements in intra- and extra-intestinal symptoms, and bifidogenic and butyrogenic effects.

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