Guar gum as a microbially degradable component for an oral colon delivery system based on a combination strategy: formulation and in vitro evaluation.

Oral colon delivery has widely been pursued exploiting naturally occurring polysaccharides degraded by the resident microbiota. However, their hydrophilicity may hinder the targeting performance. The aim of the present study was to manufacture and evaluate a double-coated delivery system leveraging intestinal microbiota, pH, and transit time for reliable colonic release. This system comprised a tablet core, a hydroxypropyl methylcellulose (HPMC) inner layer and an outer coating based on Eudragit® S and guar gum. The tablets were loaded with paracetamol, selected as a tracer drug because of the well-known analytical profile and lack of major effects on bacterial viability. The HPMC and Eudragit® S layers were applied by film-coating. Tested for in vitro release, the double-coated systems showed gastroresistance in 0.1 N HCl followed by lag phases of consistent duration in phosphate buffer pH 7.4, imparted by the HPMC layer and synergistically extended by the Eudragit® S/guar gum one. In simulated colonic fluid with fecal bacteria from an inflammatory bowel disease patient, release was faster than in the presence of ß-mannanase and in control culture medium. The bacteria-containing fluid was obtained by an experimental procedure making multiple tests possible from a single sampling and processing run. Thus, the study conducted proved the feasibility of the delivery system and ability of guar gum to trigger release in the presence of colon bacteria without impairing the barrier properties of the coating. Finally, it allowed an advantageous simulated colonic fluid preparation procedure to be set up, reducing the time, costs, and complexity of testing and enhancing replicability.

A Thermophile-Fermented Compost Modulates Intestinal Cations and the Expression of a Juvenile Hormone-Binding Protein Gene in the Female Larvae of Hercules Beetle Dynastes hercules (Coleoptera: Scarabaeidae).

The Hercules beetle larvae grow by feeding on humus, and adding a thermophile-fermented compost to the humus can upregulate the growth of female larvae. In this study, the effects of compost on the intestinal environment, including pH, cation concentrations, and organic acid concentrations of intestinal fluids, were investigated, and the RNA profile of the fat body was determined. Although the total intestinal potassium ions were similar between the larvae grown without compost (control larvae) and those with compost (compost larvae), the proportion of potassium ions in the midgut of the compost larvae drastically increased. In the midgut, an unidentified organic acid was the most abundant, and its concentration increased in the compost larvae. Transcriptome analysis showed that a gene encoding hemolymph juvenile-binding protein (JHBP) was expressed in the compost female larvae and not in the control female larvae. Expression of many genes involved in the defensive system was decreased in the compost female larvae. These results suggest that the female-specific enhancement of larval growth by compost was associated with the increased JHBP expression under conditions in which the availability of nutrition from the humus was improved by an increase in potassium ions in the midgut.

Effect of sprouted whole pearl millet on growth performance, intestinal development, bacterial count, and blood indices of broiler chickens.

This study investigated the effects of varying levels of sprouted whole grain pearl millet (SPM) on growth performance, intestinal morphology, microbial count, and blood indices of broiler chickens. A maize-soybean meal basal diet was formulated and fed to broiler chickens as starter (0 to 21 d) and finisher (22 to 42 d) diets. The diets comprised of 0%, 25%, 50%, 75%, and 100% of SPM incorporated as whole grain. On 0 d, 180 unsexed broiler chickens were allocated to experimental diets in a completely randomized design. Each treatment was replicated three times; each replicate had 12 chicks. All diets were isonitrogenous and isocaloric to meet the nutrient requirements of broiler chickens. Diets and water were provided ad libitum for 42 d. Results showed that the body weight gain (BWG) of broiler chickens on SPM compared favorably with those on the control diet. BWG showed trends in increment (P < 0.10) while FCR showed decreased trends (P < 0.10) with partial inclusion of SPM at 42 d and 0 to 42 d. The drumstick weight showed quadratic effect (P = 0.044) while the wing weight showed linear effect (P = 0.047) to treatment diets at 21 d. The liver weights of broiler chickens showed linear response (P = 0.018) at 21 d and (P = 0.004) at 42 d to SPM inclusion in diets. Sprouted whole PM consistently increased low-density lipoprotein concentration and mean corpuscular hemoglobin concentration (P < 0.05). Length and weight of small intestine and ceca showed decreasing trends on SPM levels in the treatment diets. Digesta pH assessment revealed that pH in the crop was lower (P < 0.05) on partial SPM while pH in proventriculus was reduced (P < 0.05) with inclusion of SPM in treatment diets. Lactobacilli count decreased linearly (P = 0.010) with SPM inclusion. This study suggests that SPM could be used as an alternative source of energy in production of broiler chickens. Therefore, partial replacement of maize with SPM in broiler diet had no negative effect on performance, physiological status, and overall health of broiler chickens.

Stability of Methylphenidate under Various pH Conditions in the Presence or Absence of Gut Microbiota.

Methylphenidate is one of the most widely used oral treatments for attention-deficit/hyperactivity disorder (ADHD). The drug is mainly absorbed in the small intestine and has low bioavailability. Accordingly, a high interindividual variability in terms of response to the treatment is known among ADHD patients treated with methylphenidate. Nonetheless, very little is known about the factors that influence the drug's absorption and bioavailability. Gut microbiota has been shown to reduce the bioavailability of a wide variety of orally administered drugs. Here, we tested the ability of small intestinal bacteria to metabolize methylphenidate. In silico analysis identified several small intestinal bacteria to harbor homologues of the human carboxylesterase 1 enzyme responsible for the hydrolysis of methylphenidate in the liver into the inactive form, ritalinic acid. Despite our initial results hinting towards possible bacterial hydrolysis of the drug, up to 60% of methylphenidate is spontaneously hydrolyzed in the absence of bacteria and this hydrolysis is pH-dependent. Overall, our results indicate that the stability of methylphenidate is compromised under certain pH conditions in the presence or absence of gut microbiota.

Measurement of the Intestinal pH in Mice under Various Conditions Reveals Alkalization Induced by Antibiotics.

The intestinal pH can greatly influence the stability and absorption of oral drugs. Therefore, knowledge of intestinal pH is necessary to understand the conditions for drug delivery. This has previously been measured in humans and rats. However, information on intestinal pH in mice is insufficient despite these animals being used often in preclinical testing. In this study, 72 female ICR mice housed in SPF (specific pathogen-free) conditions were separated into nine groups to determine the intestinal pH under conditions that might cause pH fluctuations, including high-protein diet, ageing, proton pump inhibitor (PPI) treatment, several antibiotic treatment regimens and germ-free mice. pH was measured in samples collected from the ileum, cecum and colon, and compared to control animals. An electrode, 3 mm in diameter, enabled accurate pH measurements with a small amount of gastrointestinal content. Consequently, the pH values in the cecum and colon were increased by high-protein diet, and the pH in the ileum was decreased by PPI. Drastic alkalization was induced by antibiotics, especially in the cecum and colon. The alkalized pH values in germ-free mice suggested that the reduction in the intestinal bacteria caused by antibiotics led to alkalization. Alkalization of the intestinal pH caused by antibiotic treatment was verified in mice. We need further investigations in clinical settings to check whether the same phenomena occur in patients.

Lessons learnt from MyCyFAPP Project: Effect of cystic fibrosis factors and inherent-to-food properties on lipid digestion in foods.

Unveiling mechanisms underpinning nutrient digestion has raised interest in the field of medical sciences for their potential application in clinical treatments. In the case of Cystic Fibrosis (CF), there exists an urgent need for understanding food lipid digestion to establish a criterion to adjust the dose of pancreatic enzyme supplements; patients have to take the supplements to allow digestion, given the associated exocrine pancreatic insufficiency (EPI). The aim of MyCyFAPP Project was to establish an evidence-based method to adjust pancreatic enzyme replacement therapy. To solve this challenge, the still unexplored field of real foods digestion had to be addressed. This review paper provides a description of the static in vitro digestion model that simulated different EPI intestinal conditions to conduct an extensive experimental work with 52 foods. Then, a summary of the data modelling that allowed for establishing a dosing criterion for enzyme supplements is provided. Following, by means of examples, an overview of the main findings related to the new knowledge generated in the field of lipid digestion in real foods is discussed, including the role of the inherent-to-food and the host factors affecting lipolysis. Finally, a discussion about the translation of the generated results in the lab to the clinical treatment of CF concludes with the lessons learnt from conducting this study.

Beneficial effects of probiotic and phytobiotic as growth promoter alternative to antibiotic for safe broiler production.

OBJECTIVES: Drug residues in poultry products could lead to the development of antibiotic-resistant bacteria as in any living animal and human alike. Extensive use of antibiotics in animals to promote growth rate, increase feed efficiency, and for prevention of intestinal infections has led to the development of resistant bacteria in the gastrointestinal tract. The present study was conducted to evaluate the effects of biological supplementation of probiotic, phytobiotic, and their combination over antibiotic on growth performance, microbial load, and hematological parameters in Broiler. MATERIALS AND METHODS: Sixty-five broiler chicken were divided into five groups (12 birds in each group), namely, group A (basal diet), group B (antibiotic, Renamycin 100®), group C (phytobiotic, Galibiotic®), group D (probiotic, Bio-Top®), and group E (combination, Galibiotic®+Galibiotic®) and five were sacrificed for baseline data on day 0. RESULTS: Average final live weight gain was highest in group D (probiotic) than other groups. The feed conversion ratio was highest in group A and lowest in the probiotic group (group C). Blood samples were collected on 14th and 28th day for hematological studies. The mean hematology values regarding the total erythrocyte count, hemoglobin concentration, packed cell volume, and erythrocyte sedimentation rate differed significantly (p < 0.05) among groups. The pH of all the treatment groups was significantly decreased compared to the control group (p < 0.05) where group C was significantly (p < 0.05) lower than all other groups. Highest total viable cell count was observed in control (group A) and total coliform count in phytobiotic (group C) was significantly lower than in other treatment groups (p < 0.05). CONCLUSION: It may conclude that biological supplements have a significant positive impact on growth performance, hematological parameters, and gut microbial load in broiler chicken of which the probiotic showing the best effects. Supplementation of probiotic in feed could be one of the best candidates as an alternative to antibiotics as growth promoter for safe broiler production.

In vitro digestion models to assess lipolysis: The impact of the simulated conditions of gastric and intestinal pH, bile salts and digestive fluids.

In vitro digestion models are a valid methodology to study nutrient hydrolysis by simulating standard physiological gastrointestinal conditions. However, there are pathologies in which some conditions are affected, which should be considered in the design of an in vitro digestion study. Our work aims at elucidating the role of different gastrointestinal conditions on lipolysis. In the context of exocrine pancreatic insufficiency, gastric pH, intestinal pH, bile salts composition, bile salts concentration, fat concentration in the digestion medium and volumetric ratio digestion fluid/food were the selected study parameters. The pH-stat method was applied to assess lipolysis extent and kinetics. Descriptive results were summarised in digestibility curves and beta regression models were used to explain the effect (odds ratio, OR) of the studied conditions on lipolysis. Additionally, distribution of emulsion droplets was measured and optical microscopy images of fat globules were taken in a selection of experimental conditions. Results showed that intestinal pH was the variable with the highest effect on lipolysis (OR 22.86, p < 0.001), followed by fat concentration in the digestion medium (OR 6.76, p < 0.001) and bile salts concentration (OR 1.56, p < 0.001). Overall, lipolysis was significantly associated with particle size (OR - 6.98, p < 0.001). We conclude that the assessment of lipolysis by means of in vitro digestion models is sensitive to the simulated gastrointestinal conditions, which should be adapted to the real physiological conditions occurring in altered health conditions.

Dose-dependent response to phytobiotic supplementation in feed on growth, hematology, intestinal pH, and gut bacterial load in broiler chicken.

OBJECTIVE: The present study was aimed to evaluate dose-dependent effects of phytobiotic (Galibiotic) supplements in feed on growth performance, hematological parameters, intestinal pH, and gut bacterial population in broiler chick. MATERIALS AND METHODS: A total of 50 ten day old broiler chicks were divided into five groups, namely, Group A as control (without galibiotic), Group B (galibiotic at 1 gm/kg feed), Group C (galibiotic at 2 gm/kg feed), Group D (galibiotic at 5 gm/kg feed), and Group E (galibiotic at 10 gm/kg feed). All the birds were reared for 42 days and samples were collected before and after sacrifice. RESULTS: Live body weights showed no significant differences between the groups but overall feed conversion ratios (FCRs) of treatment groups were significantly low in Group E having the lowest. Blood samples collected for hematology differed significantly (p < 0.01) among the different groups. Intestinal pH was lower in treatment groups with Group E having the lowest. Cecal total viable count was highest in Group A and lowest in Group E. The cecal coliform count was low in all the treatment groups. CONCLUSION: Along with previously published report, it may conclude that the phytobiotic could be used as an alternative to antibiotics due to positive growth performance, lower FCR, carcass quality, and improved gut health of broiler chicks.

Morphophysiological study of digestive system litter-feeding termite Cornitermes cumulans (Kollar, 1832).

Termites are the major decomposers of lignocellulosic biomass on Earth and are commonly considered as biological reactor models for lignocellulose degradation. Despite their biotechnological potential, few studies have focused on the morphophysiological aspects of the termite digestive system. We therefore analyze the morphology, ultrastructure and gut luminal pH of the digestive system in workers of the litter-feeding termite Cornitermes cumulans (Blattodea: Termitidae). Their digestive system is composed of salivary glands and an alimentary canal with a pH ranging from neutral to alkaline. The salivary glands have an acinar structure and present cells with secretory characteristics. The alimentary canal is differentiated into the foregut, midgut, mixed segment and hindgut, which comprises the ileum (p1), enteric valve (p2), paunch (p3), colon (p4) and rectum (p5) segments. The foregut has a well-developed chewing system. The midgut possesses a tubular peritrophic membrane and two cell types: digestive cells with secretory and absorptive features and several regenerative cells in mitosis, both cell types being organized into regenerative crypts. The mixed segment exhibits cells rich in glycogen granules. Hindgut p1, p4 and p5 segments have flattened cells with a few apical invaginations related to mitochondria and a thick cuticular lining. Conversely, the hindgut p3 segment contains large cuboid cells with extensive apical invaginations associated with numerous mitochondria. These new insights into the morphophysiology of the digestive system of C. cumulans reveal that it mobilizes lignocellulose components as a nutritional source by means of a highly compartmentalized organization with specialized segments and complex microenvironments.

Cystic fibrosis and the role of gastrointestinal outcome measures in the new era of therapeutic CFTR modulation.

With the development of new drugs that directly affect CFTR protein function, clinical trials are being designed or initiated for a growing number of patients with cystic fibrosis. The currently available and accepted clinical endpoints, FEV1 and BMI, have limitations. The aim of this report is to draw attention to the need and the ample possibilities for the development and validation of relevant gastrointestinal clinical endpoints for scientific evaluation of CFTR modulation treatment, particularly in young children and infants. The gastrointestinal tract offers very good opportunities to measure CFTR protein function and systematically evaluate CF related clinical outcomes based on the principal clinical gastrointestinal manifestations of CF: intestinal pH, intestinal transit time, intestinal bile salt malabsorption, intestinal inflammation, exocrine pancreatic function and intestinal fat malabsorption. We present a descriptive analysis of a variety of gastrointestinal outcome measures for clinical relevance, reliability, validity, responsiveness to interventions, feasibility in particular in young children and the availability of reference values.