Short-Chain Fatty Acid Butyrate Induces Cilia Formation and Potentiates the Effects of HDAC6 Inhibitors in Cholangiocarcinoma Cells

Cholangiocarcinoma (CCA) is a deadly form of liver cancer with limited therapeutic approaches. The pathogenesis of CCA involves the loss of primary cilia in cholangiocytes, an important organelle that regulates several key cellular functions including the regulation of cell polarity, growth, and differentiation, by a mechanism involving increased expression of deacetylases like HDAC6 and SIRT1. Therefore, cilia restoration may represent an alternative and novel therapeutic approach against CCA. Butyrate is produced by bacterial fermentation of fibers in the intestine and has been shown to inhibit SIRT1, showing antitumor effects on various cancers. Herein, we investigated the role of butyrate on CCA cell proliferation, migration, and EMT and evaluated the synergistic effects with specific HDAC6 inhibition. When CCA cells, including HuCCT1 and KMCH, were treated with butyrate, the cilia formation and acetylated-tubulin levels were increased, while no significant effects were observed in normal human cholangiocytes. Butyrate treatment also depicted reduced cell proliferation in HuCCT1 and KMCH cells, but on the other hand, it affected cell growth of the normal cholangiocytes only at high concentrations. In HuCCT1 cells, spheroid formation and cell migration were also halted by butyrate treatment. Furthermore, we found that butyrate augmented the previously described effects of HDAC6 inhibitors on CCA cell proliferation and migration by reducing the expression of CD44, cyclin D1, PCNA, Zeb1, and Vimentin. In summary, butyrate targets cancer cell growth and migration and enhances the anti-cancer effects of HDAC6 inhibitors in CCA cells, suggesting that butyrate may have therapeutic effects in CCA and other ciliopathies.

Optimised Method for Short-Chain Fatty Acid Profiling of Bovine Milk and Serum

Short-chain fatty acids (SCFA, C2-C5) in milk and serum are derived from rumen bacterial fermentation and, thus, have the potential to be used as biomarkers for the health status of dairy cows. Currently, there is no comprehensive and validated method that can be used to analyse all SCFAs in both bovine serum and milk. This paper reports an optimised protocol, combining 3-nitrophenylhydrazine (3-NPH) derivatisation and liquid chromatography-mass spectrometry (LC-MS) analysis for quantification of SCFA and β-hydroxybutyric acid (BHBA) in both bovine milk and bovine serum. This method is sensitive (limit of detection (LOD) ≤ 0.1 µmol/L of bovine milk and serum), accurate (recovery 84–115% for most analytes) and reproducible (relative standard deviation (RSD) for repeated analyses below 7% for most measurements) with a short sample preparation step. The application of this method to samples collected from a small cohort of animals allowed us to reveal a large variation in SCFA concentration between serum and milk and across different animals as well as the strong correlation of some SCFAs between milk and serum samples.

Improving Oil Recovery of the Heterogeneous Low Permeability Reservoirs by Combination of Polymer Hydrolysis Polyacrylamide and Two Highly Biosurfactant-Producing Bacteria

Polymer hydrolysis polyacrylamide and microbes have been used to enhance oil recovery in many oil reservoirs. However, the application of this two-method combination was less investigated, especially in low permeability reservoirs. In this work, two bacteria, a rhamnolipid-producing Pseudomonas aeruginosa 8D and a lipopeptide-producing Bacillus subtilis S4, were used together with hydrolysis poly-acrylamide in a low permeability heterogeneous core physical model. The results showed that when the two bacterial fermentation liquids were used at a ratio by volumeof 1:3 (v:v), the mixture showed the optimal physicochemical properties for oil-displacement. In addition, the mixture was stable under the conditions of various temperature (20–70 °C) and salinity (0–22%). When the polymer and bacteria were mixed together, it had no significant effects in the viscosity of polymer hydrolysis polyacrylamide and the viability of bacteria. The core oil-displacement test displayed that polymer hydrolysis polyacrylamide addition followed by the bacterial mixture injection could significantly enhance oil recovery. The recovery rate was increased by 15.01% and 10.03%, respectively, compared with the sole polymer hydrolysis polyacrylamide flooding and microbial flooding. Taken together, these results suggest that the strategy of polymer hydrolysis poly-acrylamide addition followed by microbial flooding is beneficial for improving oil recovery in heterogeneous low permeability reservoirs.

A Review of The Contribution of Gut-Dependent Microbiota Derived Marker, Trimethylamine N-oxide (TMAO), in Coronary Artery Disease

Coronary artery disease (CAD) has a high prevalence and one of the principal drivers of mortality worldwide. Therefore, there is a requirement to develop sensitive diagnostic biomarkers, disease progression control and therapeutic stratification in order to keep a check on the disease rate. Atherosclerosis is a systemic disease, the main cause of heart disease, is associated with hyperlipidemia and lipid oxidation and has always been a common single leading cause of death in well-developed countries. In the attempts to study CAD and the causative agents for the disease, a metabolite circulating in the plasma termed trimethylamine-N-oxide (TMAO) has been found out to be an independent risk factor that increases CAD risk. The use of a metabolomic approach has proven useful in the recent past, as it can aid in the identification and quantification of several metabolites that play a crucial role for diagnosis and exploring therapeutic targets. TMAO is majorly synthesized by a process which involves the bioconversion of gut microbiota and hepatic flavin monooxygenases (FMOs) from nutrient-containing dietary trimethylamine (TMA). TMA is synthesized by gut bacterial fermentation from the components present in meat such as phosphatidylcholine (PC), betaine, choline, and L-carnitine. It can accentuate the process of atherosclerosis through the novel meta-organismal metabolic pathway. TMAO leads to atherogenesis by increasing vascular inflammation, reducing vascular functions and disrupting cholesterol homeostasis at various levels. This review article attempts to summarize the pool of evidence collected on the microbiota-dependent TMAO and its association with atherosclerosis. We performed literature search with Medline, PubMed, and Google Scholar, on “TMAO in CAD”, “metabolites in CAD” and “TMAO in other diseases” from the year 1990 to 2020. Although the circulatory TMAO has been identified as an independent marker for CAD, there is still no conclusive evidence to justify its role as a routine marker for CAD diagnosis. Future research must clarify the mechanisms which underpin these complex associations to determine if there is a causal link exists between TMAO and CAD.

Impact of Long-Term Storage on Alfalfa Leaf and Stem Silage Characteristics

Fractionation of alfalfa (Medicago sativa L.) into leaves and stems can reduce cutting frequency while producing a high-value feedstuff. A difficulty with fractionation is the higher moisture content at ensiling due to the inability to wilt leaves without substantial dry matter loss or financial cost. To evaluate whether the silage quality of fractionated alfalfa is competitive with conventional methods under long-term storage conditions, high-moisture leaves (250–280 g·kg−1 dry matter, DM) and stems (190–240 g·kg−1 DM) were ensiled for 21 and 350 days and compared to wilted whole-plant silage for two alfalfa developmental stages. At 21 days, leaf and whole-plant silage fermentation characteristics indicated suitable lactic acid bacterial fermentation through decreased pH, high lactic acid–to-acetic acid ratio, and a lack of clostridial fermentation. At 350 days, leaf silage fermentation and nutritional quality decreased due to sustained proteolysis, but true protein still exceeded that of whole-plant silage. High-moisture stem fractions fermented poorly; at 21 days, stage 3 stems had significant amounts of butyric acid, while stage 5 stems became clostridial at 350 days. Long-term storage of high-moisture leaf silages can produce good-quality silage despite exceeding moisture contents typically recommended for alfalfa, while wilting is required for stem silages.

Impact of zinc oxide, benzoic acid and probiotics on the performance and cecal microbiota of piglets

Background Intestinal health remains a key factor in animal production because it is essential for digestion, absorption and bacterial fermentation. Feed additives have been used to attenuate the weaning stress such as Zinc Oxide (ZnO) and benzoic acid (C7H6O2). The objective of this study was to evaluate the impact of of benzoic acid and probiotics (BA + P) on performance, diarrhea and cecal microbiota of piglets in the nursery phase (23 to 65 days). Results One hundred and sixty weaned piglets with an initial weight of 6.335 ± 0.698 kg and 23 days of age were submitted to four treatments: supplementation with 2500 ppm of Zinc oxide (ZnO), supplementation with a commercial blend of benzoic acid and probiotics (Bacillus licheniformis, Bacillus subtilis and Enterococcus faecium NCIMB 10415; Vevogut P®) (BA + P), supplementation with Zinc oxide plus benzoic acid and probiotics (ZnO + BA + P), and controls receiving only the basal diet without any supplementation. At 65 days of age, 32 piglets (n = 8 per treatment) were slaughtered for the evaluation of the cecal microbiota. Supplementation with ZnO and BA + P were associated with better feed conversion (P < 0.05) in the early stage (23 to 49 days) and with an improvement in all performance parameters over the entire experimental period. The occurrence of diarrhea was lower (P < 0.05) in the BA + P group. The 4 most abundant phyla along with unclassified bacteria represented 93% of all sequences. Firmicutes dominated the cecal microbiota of all groups, followed by Bacteroidetes. Richness represented by the observed number of genera and by the Chao index were statistically lower in ZnO and ZnO + BA + P supplemented animals compared to controls. The beta diversity analysis that compares similarities between bacterial communities demonstrated formation of two distinct clusters containing samples with and without supplementation with ZnO, confirming a strong influence of ZnO on the intestinal microbiota. Conclusion The use of Benzoic acid with probiotics yields similar performance results with lower impact on the gut microbiota compared to ZnO, and it should be considered as a potential alternative in swine production.

Postbiotics as Dynamic Biological Molecules for Antimicrobial Activity: A Mini-Review

Postbiotics, products, or metabolites secreted by living probiotic bacteria like teichoic acids, peptides, enzymes, peptidoglycans, polysaccharides, organic acids, and external cell proteins are said to be produced during the bacterial fermentation process. However, postbiotics may provide immunization, antioxidant, prevents inflammation, low cholesterolemic, antimicrobial, antagonistic obesity, contrast hypertensive, and diabetic retinopathy impacts. In the current review, we attempt to display the antimicrobial performance of postbiotics. In this regard, we considered microbial strains used as postbiotic sources and postbiotics as antimicrobial agents in food products. All databases such as Science Direct, Scopus, Pub Med, and Google Scholar were examined using the following keywords: “postbiotics”, “Antimicrobial activity”, “Anti-inflammatory”, and “Low cholesterolemic”. Further studies demonstrated that probiotics are fed special forms of fiber (prebiotic) molecules, indicating substances known as postbiotics. Furthermore, short-chain fatty acids (SCFAs) like acetate, propionate, and butyrate are among the comprehensively investigated postbiotics. The extraction and purification of these compounds are carried out by means of dialysis, centrifugation, and freeze-drying techniques. According to the gained results, postbiotics assist in improving host health by increasing certain physiological functions. Furthermore, postbiotics can be used to increment the useful lifetime of different foods, like dairy products. It has also been shown that manually adding postbiotics to such products prevents the growth and proliferation of molds and thus the spoilage caused by them. This inhibitory effect indicates the antimicrobial properties of these compounds. Finally, we will see significant advances in the biological preservation of products, especially in the food industry.

Potential Association Between Dietary Fibre and Humoral Response to the Seasonal Influenza Vaccine

Influenza vaccination is an effective public health measure to reduce the risk of influenza illness, particularly when the vaccine is well matched to circulating strains. Notwithstanding, the efficacy of influenza vaccination varies greatly among vaccinees due to largely unknown immunological determinants, thereby dampening population-wide protection. Here, we report that dietary fibre may play a significant role in humoral vaccine responses. We found dietary fibre intake and the abundance of fibre-fermenting intestinal bacteria to be positively correlated with humoral influenza vaccine-specific immune responses in human vaccinees, albeit without reaching statistical significance. Importantly, this correlation was largely driven by first-time vaccinees; prior influenza vaccination negatively correlated with vaccine immunogenicity. In support of these observations, dietary fibre consumption significantly enhanced humoral influenza vaccine responses in mice, where the effect was mechanistically linked to short-chain fatty acids, the bacterial fermentation product of dietary fibre. Overall, these findings may bear significant importance for emerging infectious agents, such as COVID-19, and associated de novo vaccinations.

Production and Escalation of L-Lysine via Bacterial Fermentation Utilizing Streptococcus Sp

Background and AimsThe importance of L-lysine as an essential amino acid in the nutrition of human beings has made it desirable supplement of the diet in recent years. It can be produced in different ways among them fermentation is the most economical and practical means of producing lysine. In this method low temperature, low pressure and low-cost carbon sources are used and a biological form of lysine (L-lysine) is produced. MethodsIn the present study, the production of L-lysine was achieved through fermentation developed from locally isolated bacterial strains. In total, twenty-nine (29) bacterial strains were isolated and tested using paper chromatographic technique. Six different parameters for optimization were scrutinized for improved bacterial growth and significant yield of lysine was obtained using selected strains.Key ResultsFor Streptococcus sp. molasses media with vitamins (w) formed 24.4 g/L, 40º C generated 24.4 g/L, addition of 1mM solution of metal ion (Mg) yielded 20.4 g/L, pH 6.5 delivered 6 g/L, fermentation period of 96 hours engendered 24.4 g/L, and 0.3 mL of inoculum results in 9.2 g/L of lysine. ConclusionsLaboratory scale production of L-lysine was carried out using 1 L Erlenmeyer flask. For Streptococcus sp. 23.4 g/L of lysine was produced after optimized conditions.

Metabolism of Selected 2-Arylbenzofurans in a Colon In Vitro Model System

2-arylbenzofurans represent a small group of bioactive compounds found in the plant family Moraceae. As it has not been investigated whether these substances are stable during passage through the gastrointestinal tract, their biological effects may be altered by the metabolism of intestinal microbiota or cells. The aim of the present study was to investigate and compare mulberrofuran Y (1), moracin C (2), and mulberrofuran G (3) in an in vitro model of human intestinal bacterial fermentation and in an epithelial model using the Caco-2 cell line. The analysis of compounds by LC-MS-Q-TOF showed sufficient stability in the fermentation model, with no bacterial metabolites detected. However, great differences in the quantity of permeation were observed in the permeability assay. Moreover, mulberrofuran Y (1) and moracin C (2) were observed to be transformed into polar metabolites by conjugation. Among the test compounds, mulberrofuran Y (1) was mostly stable and accumulated in endothelial cells (85.3%) compared with mulberrofuran G (3) and moracin C (2) (14% and 8.2%, respectively). Thus, only a small amount of mulberrofuran Y (1) was conjugated. Moracin C (2) and mulberrofuran G (3) were metabolized almost completely, with only traces of the unchanged molecule being found on the apical and cellular sides of the system. Only conjugates of mulberrofuran Y (1) and moracin C (2) were able to reach the basolateral side. Our results provide the basic description of bioavailability of these three compounds, which is a necessary characteristic for final evaluation of bio-efficacy.