Influence of feeding practices in the composition and functionality of infant gut microbiota and its relationship with health.

Establishing the infant's gut microbiota has long-term implications on health and immunity. Breastfeeding is recognized as the best practice of infant nutrition in comparison with formula feeding. We evaluated the effects of the primary feeding practices by analyzing the infant growth and the potential association with gut diseases. A cross-sectional and observational study was designed. This study included 55 mothers with infants, who were divided according to their feeding practices in breastfeeding (BF), formula feeding (FF), and combined breast and formula feeding (CF). Anthropometric measurements of the participants were recorded. Additionally, non-invasive fecal samples from the infants were collected to analyze the microbiota by sequencing, immunoglobulin A (IgA) concentration (ELISA), and volatile organic compounds (gas chromatography with an electronic nose). Results showed that the microbiota diversity in the BF group was the highest compared to the other two groups. The IgA levels in the BF group were twice as high as those in the FF group. Moreover, the child´s growth in the BF group showed the best infant development when the data were compared at birth to the recollection time, as noted by the correlation with a decreased concentration of toxic volatile organic compounds. Interestingly, the CF group showed a significant difference in health status when the data were compared with the FF group. We conclude that early health practices influence children's growth, which is relevant to further research about how those infants' health evolved.

Production and purification of bacterial membrane vesicles for biotechnology applications: Challenges and opportunities.

Bacterial membrane vesicles (BMVs) are bi-layered nanostructures derived from Gram-negative and Gram-positive bacteria. Among other pathophysiological roles, BMVs are critical messengers in intercellular communication. As a result, BMVs are emerging as a promising technology for the development of numerous therapeutic applications. Despite the remarkable progress in unveiling BMV biology and functions in recent years, their successful isolation and purification have been limited. Several challenges related to vesicle purity, yield, and scalability severely hamper the further development of BMVs for biotechnology and clinical applications. This review focuses on the current technologies and methodologies used in BMV production and purification, such as ultracentrifugation, density-gradient centrifugation, size-exclusion chromatography, ultrafiltration, and precipitation. We also discuss the current challenges related to BMV isolation, large-scale production, storage, and stability that limit their application. More importantly, the present work explains the most recent strategies proposed for overcoming those challenges. Finally, we summarize the ongoing applications of BMVs in the biotechnological field.

Assessment of Potential Probiotic and Synbiotic Properties of Lactic Acid Bacteria Grown In Vitro with Starch-Based Soluble Corn Fiber or Inulin.

This research is aimed to search for suitable probiotic plus prebiotic combinations for food applications. Sixteen bacteria were tested for resistance to low pH, bile salts and antibiotics, and their adhesion to Caco-2 cells, in order to select potential probiotics. Then, two bacteria were selected to study short chain fatty acids production in a starch-based soluble corn fiber or inulin media. Lactiplantibacillus plantarum V3 and L. acidophilus La3 manifested the best probiotic features with a remarkable adhesion ability (23.9% and 17.3%, respectively). Structural differences between fibers have an impact on how each one is metabolized, both in their capacity of being easily fermented and in the short chain fatty acids profile obtained: L. acidophilus La3 in inulin fermentation yielded the highest total short chain fatty acids (85.7 mMol/L), and, in starch-based soluble corn fiber fermentation, yielded the highest butyric acid content (0.31 mMol/L). This study provides valuable information for future design of synbiotics for food applications.

Fish oil and probiotics supplementation through milk chocolate improves spatial learning and memory in male Wistar rats.

Background: Cognition and brain function is critical through childhood and should be improved with balanced diets. Incorporating bioactive ingredients such as omega-3 polyunsaturated fatty acids (ω3 PUFAs) and probiotics into food formulations could be used as an approach to improve cognitive function. This study evaluated the effects on cognitive capacity of complementing rodent diets with chocolate, by itself and in combination with ω3 PUFAs from fish oil and probiotics. Methods: Spatial learning and memory in the rats were determined by the Barnes maze test in short- and long-term memory. Samples from the cecum were obtained to assess microbial counts (Lactobacillus, Bifidobacterium, Enterobacteriaceae, and total bacteria), and brains were recovered to analyze the neural morphology of the tissues. Also, glucose, brain weights, and epididymal tissue were analyzed. Results: The combination of chocolate with fish oil and probiotics improved the memory of rats compared to the result of each bioactive compound when evaluated separately. Treatments did not affect sugar level, epididymal adipose tissue, or brain weight. On the other hand, consuming probiotics alone or in combination with chocolate decreased Enterobacteria counts, while Lactobacillus and Bifidobacteria counts were not affected. Neural morphological analysis showed that combining chocolate with probiotics and ω3 PUFAs increased the number of neurons in the hippocampal CA1 and CA3 regions. Conclusion: Chocolate added with probiotics and ω3 PUFAs improved spatial memory and learning in the studied model.

Spray dried lactobacilli maintain viability and feruloyl esterase activity during prolonged storage and under gastrointestinal tract conditions.

The use of lactobacilli with feruloyl esterase (FE) activity in the development of functional foods has gained considerable interest in recent years. Microencapsulation of FE-producing bacteria to facilitate their incorporation into food is a challenge. The aim of this study was to evaluate survival and maintenance of FE activity during storage at 4 °C and under simulated gastrointestinal tract (GIT) conditions of microcapsules of FE-producing Lactobacillus (Lb.) strains obtained by spray drying. Lb. fermentum CRL1446 and Lb. johnsonii CRL1231 powders maintained viability at concentrations ≥ 106 CFU/g (minimum probiotic dose) when stored at 4 °C for 12 months. Lb. acidophilus CRL1014 powders were only able to maintain ≥ 106 CFU/g during 4 months of storage. FE activity was conserved in three microencapsulated strains evaluated, an increase of specific activity being observed until month 12 of storage. Powders of the three strains incubated under GIT conditions maintained their viability (≥ 106 CFU/g), but specific FE activity was only detected in Lb. fermentum and Lb. johnsonii powders (0.80-0.83 and 0.21-0.56 U/mg, respectively). CRL1446 and CRL1231 microcapsules were able to resist prolonged storage and GIT conditions, retaining FE activity and preserving their probiotic potential and could be incorporated into functional foods.

Chocolate as Carrier to Deliver Bioactive Ingredients: Current Advances and Future Perspectives.

Consumer demand for healthier foods with improved taste and convenience has urged the food industry to develop functional foods added with bioactive ingredients that can supplement basic nutrition (food supplement) or exert a pharmacological effect (nutraceuticals). Chocolate could be used as an ideal carrier to deliver bioactive ingredients, mainly due to its high acceptability by consumers. However, a drawback of using chocolate as functional food is its high sugar content, which impedes its commercialization with the diabetic population. Therefore, there is need to develop sugar-free chocolate formulations added with bioactive ingredients. Nevertheless, sugar replacement and bioactive ingredients addition is a major technological challenge that affects texture, rheology, and sensory properties of chocolate. This review is designed as a practical guide for researchers and food industries to develop the next generation of functional chocolates. Different functional chocolate formulations, including sugar-free, are reviewed as potential carriers for the delivery of bioactive compounds. The physicochemical properties and sensory acceptability of the functional chocolates presented are also highlighted. Finally, future perspectives, such as the use of nanotechnology to improve the bioaccessibility and bioavailability of active ingredients, as well as the need for clinical trials to validate the pharmacological effect of functional chocolates, are also discussed.

Sugar-Free Milk Chocolate as a Carrier of Omega-3 Polyunsaturated Fatty Acids and Probiotics: A Potential Functional Food for the Diabetic Population.

Chocolate is an adequate matrix to deliver bioactive ingredients. However, it contains high sugar levels, one of the leading causes of chronic degenerative diseases. This work aimed to evaluate the effects of milk chocolate reformulation with alternative sugar sweeteners (Sw; isomalt + stevia), probiotics (Prob), and ω-3 polyunsaturated fatty acids (PUFAs) on its physicochemical properties and consumers' acceptability. Lactobacillus plantarum 299v (L. p299v) and Lactobacillus acidophilus La3 (DSMZ 17742) were added as Prob strains, and fish oil (FO) was added as the source of ω-3 PUFAs. Prob addition resulted in chocolates with >2 × 107 colony forming unit (CFU) per serving size (12 g). Except for Prob, aw values of all treatments were <0.46. Sw and Sw + Prob presented the nearest values to the control in hardness, whereas Sw without FO increased fracturability. FO, Sw + FO, and Sw + Prob + FO contained 107.4 ± 12.84, 142.9 ± 17.9, and 133.78 ± 8.76 mg of ω-3 PUFAs per chocolate, respectively. Prob + FO increased the resistance of chocolate to shear stress, while Sw + FO showed a similar flow behavior to the control. The consumers' acceptability of Sw + Prob chocolate was adequate, while Sw + Prob + FO had higher acceptability than Prob + FO. Health benefits of reformulated milk chocolates requires further assessment by in vitro, in vivo and clinical studies.

The complex relationship between metabolic syndrome and sweeteners.

Metabolic syndrome is a multifactorial disorder originating from central obesity through a high caloric intake and a sedentary lifestyle. Metabolic syndrome increases the risk of type 2 diabetes (T2D) disease, converting it to one of the costliest chronic diseases, which reduces life quality. A strategy proposed by the food industry to reduce this problem is the generation of low-caloric products using sweeteners, which are compounds that can substitute sucrose, given their sweet taste. For many years, it was assumed that sweeteners did not have a relevant interaction in metabolism. However, recent studies have demonstrated that sweeteners interact either with metabolism or with gut microbiota, in which sweet-taste receptors play an essential role. This review presents an overview of the industrial application of most commonly consumed sweeteners. In addition, the interaction of sweeteners within the body, including their absorption, distribution, metabolism, gut microbiota metabolism, and excretion is also reviewed. Furthermore, the complex relationship between metabolic syndrome and sweeteners is also discussed, presenting results from in vivo and clinical trials. Findings from this review indicate that, in order to formulate sugar-free or noncaloric food products for the metabolic syndrome market, several factors need to be considered, including the dose, proportions, human metabolism, and interaction of sweeteners with gut microbiota and sweet-taste receptors. More clinical studies, including the metabolic syndrome, are needed to better understand the interaction of sweeteners with the human body, as well as their possible effect on the generation of dysbiosis.

Assessment of the bacterial diversity of agave sap concentrate, resistance to in vitro gastrointestinal conditions and short-chain fatty acids production.

Low bacterial diversity in the gut has been associated with the development of several diseases. Agave sap concentrate (ASC) is obtained from the thermal treatment of the fresh sap called "aguamiel", an artisanal Mexican food. In this study, we assessed the microbial diversity from three different ASC producing regions in Mexico using high-throughput sequencing of the 16S rRNA gene and evaluated their resistance to an in vitro gastrointestinal process as well as their ability to produce short-chain fatty acids (SCFA). Seven phyla and 120 genera were detected in ASC samples; Firmicutes had the highest relative read abundance at the phylum level, whereas Bacillus was the most abundant genus. Bacterial diversity at phylum and genus levels was highly dependent on the region where ASC was produced. The microbiota from a selected sample was resistant to low pH conditions, bile salts and intestinal enzymes. Moreover, bacteria were able to survive and grow in the colonic environment. SCFA production was comparable with that observed for a well-known probiotic, Lactobacillus plantarum 299v, that was used as control. These findings demonstrate that ASC contains a bacterial ecosystem with potential probiotic benefits.

Physicochemical Properties and Sensory Acceptability of a Next-Generation Functional Chocolate Added with Omega-3 Polyunsaturated Fatty Acids and Probiotics.

In this study, a milk chocolate formulation was developed to serve as vehicle of Omega-3 (ω3) polyunsaturated fatty acids (PUFAs) and probiotics (L. plantarum 299v and L. rhamnosus GG). Fish oil (FO) was incorporated in chocolate as a source of ω3 PUFAs. Probiotics (Prob) and FO were added during tempering, obtaining chocolates with 76.0 ± 5.2 mg (FO1) or 195.8 ± 6.5 mg (FO2) of ω3 PUFAs, and >1 × 106 CFU of Prob per chocolate portion (12 g). The physicochemical properties (rheological analysis, texture, surface instrumental color, aw, and fatty acid profile), and sensory acceptability of the formulations were determined. Prob and FO generated a decrease in L* and white index (WI) values. Except for Prob + FO2, all treatments showed a decrease in aw. Rheological parameters of FO1 and Prob + FO1 presented the most similar behavior as compared with the control. Prob or FO1 addition did not affect the overall consumer's acceptability of chocolate; and when both nutraceuticals were combined (Prob + FO1) the product showed adequate overall acceptability. FO2 formulations were not considered adequate to maintain physicochemical properties and sensory acceptability of chocolate. Results indicated that milk chocolate is a suitable vehicle for delivering ω3 PUFAs and Prob, which are essential to enhance cognitive development in children.

Sanitizing after fresh-cutting carrots reduces the wound-induced accumulation of phenolic antioxidants compared to sanitizing before fresh-cutting.

BACKGROUND: During the production of fresh-cut products, crops are exposed to wounding stress, and as a stress response, phenolic antioxidants are synthesized. This stress response is elicited by extracellular adenosine triphosphate, released from wounded cells and recognized by receptors of unwounded cells. The phenolic antioxidants produced as a stress response are beneficial for human health. However, a common practice in the fresh-cut industry is the application of washing/sanitizing procedures after cutting. These procedures could be highly detrimental, since they partially remove the wound signal that elicits the biosynthesis of phenolics in plants. In this study, the impact of different washing/sanitizing treatments post-shredding on the wound-induced accumulation of chlorogenic acid (CHA) in carrot was evaluated. Peeled carrots were shredded and dipped in aqueous solutions containing chlorine (100 ppm, 2 min), hydrogen peroxide (1.5%, 2 min) or water (2 min). The content of CHA in treated carrots was evaluated before and after 48 h of storage (19 ± 2 °C). RESULTS: The control carrots sanitized only before peeling and shredding showed 4000% higher content of CHA as compared with time 0 h samples. However, carrots treated with washing/sanitizing procedures post-shredding including water, chlorine and hydrogen peroxide showed a decrease in the accumulation of CHA by 46.9%, 53.6% and 89.9%, respectively. CONCLUSIONS: The results demonstrated that washing/sanitizing procedures applied after fresh-cutting are potentially detrimental to the wound-induced accumulation of health-promoting compounds during storage of fresh produce. Thus, the fresh-cut industry could consider avoiding washing procedures after cutting and implement alternative sanitizing procedures that avoid the partial removal of the wound signal, such as sanitizing only before cutting. © 2020 Society of Chemical Industry.

Postharvest Wounding Stress in Horticultural Crops as a Tool for Designing Novel Functional Foods and Beverages with Enhanced Nutraceutical Content: Carrot Juice as a Case Study.

Phenolic compounds have potential to prevent and treat chronic degenerative diseases (CDDs). A phenolic-rich carrot juice was produced by the application of wounding stress. The effects of wounding intensity, storage, peeling, blanching, filtration, and pasteurization over physicochemical, nutritional, nutraceutical, and sensory properties of carrot juice were evaluated. Juices from unpeeled carrots had 7% to 40% more minerals, 0.46 to 1.6 less °Brix, and 1.16× more titratable acidity. The carrot juice with the highest phenolic content was obtained by cutting unpeeled carrots into slices, storing them (48 hr, 15 °C), and blanching them thereafter (80 °C, 6 min; stressed unpeeled carrot juice, SUCJ). SUCJ had 3,600% more chlorogenic acid, 195% more total phenolics, and similar carotenoid content than conventional carrot juice. Sensory evaluation of SUCJ was acceptable and willingness to pay increased by providing information about health benefits. SUCJ has potential as a functional beverage that could aid in the prevention and treatment of CDDs. PRACTICAL APPLICATION: Consumers are increasingly demanding foods and beverages that are healthier, natural, safe, and GMO-free. Abiotic stresses can enhance greatly the nutraceutical content of crops without the need of genetic engineering or dangerous chemicals. These crops could be used as raw materials to produce foods and beverages of higher nutraceutical quality. An easy-to-control abiotic stress is wounding stress, which consists of mechanically damaging the plant tissue (for example, cutting). We applied wounding stress to carrot to produce a phenolic-rich carrot juice. This juice could aid in the prevention or treatment of chronic degenerative diseases.

Using a Functional Carrot Powder Ingredient to Produce Sausages with High Levels of Nutraceuticals.

In this study, a functional carrot powder (FCP) ingredient was obtained by applying wounding stress to carrot (shredding and storing for 48 hr at 15 °C) prior to dehydration (60 °C) and milling. Likewise, FCP was incorporated into sausage formulations, which were further characterized. The application of wounding stress in carrots resulted in a FCP with higher fiber (30.1%) and chlorogenic acid (798.4%) content as compared with control carrot powder (CCP). Likewise, FCP showed higher water (19%) and oil (3.9%) absorption capacity as compared with CCP. Sausage formulation with 4% (w/w) of FCP was characterized, further evaluated during storage (42 days, 4 °C), and compared with a formulation added with 4% (w/w) CCP. FCP and CCP formulations increased fiber of sausages by 72.7%, and fortified them with carotenoids, providing 30% to 40% of vitamin A daily requirements per portion (62.5 g). FCP did not affect purge loss, and sausages contained 270% and 377% more total phenolics and chlorogenic acid content as compared with CCP sausages. FCP formulation presented adequate sensory acceptability and its carotenoid and phenolic content remained stable during storage. Results indicated that FCP could be used as an ingredient in sausage formulation to increase the content of nutraceuticals without affecting its shelf-life. PRACTICAL APPLICATION: Wounding stress (by shredding) was applied to carrots prior to dehydration and milling to obtain a carrot powder with higher fiber and phenolic compounds as compared with control carrot powder (CCP). FCP showed higher water and oil absorption capacity than CCP. FCP addition to sausage formulations at 4% didn't affect purge loss, and resulted in sausages with higher phenolic compounds and dietary fiber. Furthermore, FCP formulations presented adequate sensory acceptability and its carotenoid and phenolic content remained stable during storage. FCP could be used as an ingredient in sausage formulation to increase the content of nutraceuticals without affecting its shelf-life.

Peptides and isoflavones in gastrointestinal digests contribute to the anti-inflammatory potential of cooked or germinated desi and kabuli chickpea (Cicer arietinum L.).

It is largely unknown how processing affects bioactive potential of chickpea proteins to prevent bowel inflammatory diseases. The aim was to investigate the anti-inflammatory activity of protein concentrates from germinated and cooked chickpeas (GC and CC, respectively) and its relationship with protein and isoflavone composition before and after in vitro gastrointestinal digestion and absorption. Anti-inflammatory activity of GC digests was almost 2-fold higher than CC digests (p < 0.05), which was associated to greater content of peptides, formononetin and biochanin A (p < 0.05). Anti-inflammatory activity of phenolic fraction in digests was 7-fold higher than the protein fraction (p < 0.05). The most active peptide fraction from GC digest (IC50 = 93 µg/mL) contained a total of 24 peptides derived from legumin and vicilin. In conclusion, this study stands out the potential of germinated chickpea proteins concentrates to exert anti-inflammatory effects in the lower gut which may contribute to the prevention of bowel inflammatory diseases.

High-protein diet modifies colonic microbiota and luminal environment but not colonocyte metabolism in the rat model: the increased luminal bulk connection.

High-protein diets are used for body weight reduction, but consequences on the large intestine ecosystem are poorly known. Here, rats were fed for 15 days with either a normoproteic diet (NP, 14% protein) or a hyperproteic-hypoglucidic isocaloric diet (HP, 53% protein). Cecum and colon were recovered for analysis. Short- and branched-chain fatty acids, as well as lactate, succinate, formate, and ethanol contents, were markedly increased in the colonic luminal contents of HP rats (P < 0.05 or less) but to a lower extent in the cecal luminal content. This was associated with reduced concentrations of the Clostridium coccoides and C. leptum groups and Faecalibacterium prausnitzii in both the cecum and colon (P < 0.05 or less). In addition, the microbiota diversity was found to be higher in the cecum of HP rats but was lower in the colon compared with NP rats. In HP rats, the colonic and cecal luminal content weights were markedly higher than in NP rats (P < 0.001), resulting in similar butyrate, acetate, and propionate concentrations. Accordingly, the expression of monocarboxylate transporter 1 and sodium monocarboxylate transporter 1 (which is increased by higher butyrate concentration) as well as the colonocyte capacity for butyrate oxidation were not modified by the HP diet, whereas the amount of butyrate in feces was increased (P < 0.01). It is concluded that an increased bulk in the large intestine content following HP diet consumption allows maintenance in the luminal butyrate concentration and thus its metabolism in colonocytes despite modified microbiota composition and increased substrate availability.

Protective effect of Bifidobacterium pseudocatenulatum CECT7765 against induced bacterial antigen translocation in experimental cirrhosis.

BACKGROUND & AIMS: Intervention in the gut ecosystem is considered as a potential strategy to treat liver diseases and their complications. We have evaluated the effects of Bifidobacterium pseudocatenulatum CECT7765 on bacterial translocation and the liver status in experimental cirrhosis. ANIMALS & METHODS: Liver damage was induced in Balb/c mice by weight-controlled oral administration of carbon tetrachloride. Laparotomies were performed at week 12. One week prior to laparotomy, animals received B. pseudocatenulatum CECT7765 (10(9) cfu/daily) or placebo intragastrically. All animals received Escherichia coli (10(7) cfu/single dose) intragastrically 24 hours before laparotomy. A group of naïve non-treated animals was included as control. Liver tissue specimens, mesenteric lymph nodes, intestinal content and blood were collected. Liver histology, profibrogenic genes expression, bacterial DNA translocation, serum endotoxaemia and liver cytokine levels were measured. RESULTS: Bifidobacterium pseudocatenulatum CECT7765 showed no significant effect on structural liver damage, as determined by histological evaluation, alpha-smooth muscle actin distribution, profibrogenic gene expression levels, total hydroxyproline levels and malon dialdehyde production compared with mice receiving placebo. Interestingly, bacterial DNA translocation and serum endotoxin levels were significantly decreased in mice receiving the Bifidobacterium strain compared with placebo. Gut barrier integrity markers were up-regulated in mice receiving B. pseudocatenulatum CECT7765 and quantitatively correlated with intestinal gene copy numbers of the bifidobacterial strain. Gene expression levels of several anti-inflammatory mediators were also increased in mice receiving B. pseudocatenulatum CECT7765 compared with placebo. CONCLUSION: Oral administration of B. pseudocatenulatum CECT7765 is associated with improved gut barrier integrity and shows a beneficial effect against induced bacterial antigen translocation in the CCl4 -model of cirrhosis.

Bifidobacterium CECT 7765 improves metabolic and immunological alterations associated with obesity in high-fat diet-fed mice.

OBJECTIVES: To evaluate the effects of administration of Bifidobacterium pseudocatenulatum CECT 7765 on metabolic and immune alterations in obese mice. DESIGN AND METHODS: Adult male wild-type C57BL-6 mice were fed a standard diet or high-fat diet (HFD), supplemented or not with B. pseudocatenulatum CECT 7765 for 7 weeks. The assessments included biochemical and immunological parameters, insulin resistance, glucose tolerance, histology of liver, white-adipose and intestinal tissues, immunocompetent cell functions, and microbiota-related features. RESULTS: B. pseudocatenulatum CECT 7765 reduced serum cholesterol, triglyceride, and glucose levels and decreased insulin resistance and improved glucose tolerance in obese mice. This strain reduced serum levels of leptin, interleukin (IL)-6 and monocyte chemotactic protein-1, while increased those of IL-4 in HFD-fed mice. B. pseudocatenulatum CECT7765 reduced liver steatosis and the number of larger adipocytes and number of fat micelles in enterocytes of obese mice. The strain also improved the function of macrophages and dendritic cells in relation to phagocytosis, cytokine production, and induction of T-lymphocyte proliferation. The strain administration increased bifidobacteria and reduced enterobacteria and the inflammatory properties of the gut content in HFD-fed mice. CONCLUSION: B. pseudocatenulatum CECT 7765 was shown to ameliorate both metabolic and immunological dysfunctions related to obesity in HFD-fed mice.

Bacteroides uniformis CECT 7771 ameliorates metabolic and immunological dysfunction in mice with high-fat-diet induced obesity.

BACKGROUND: Associations have been made between obesity and reduced intestinal numbers of members of the phylum Bacteroidetes, but there is no direct evidence of the role these bacteria play in obesity. Herein, the effects of Bacteroides uniformis CECT 7771 on obesity-related metabolic and immune alterations have been evaluated. METHODS AND FINDINGS: Adult (6-8 week) male wild-type C57BL-6 mice were fed a standard diet or a high-fat-diet HFD to induce obesity, supplemented or not with B. uniformis CECT 7771 for seven weeks. Animal weight was monitored and histologic, biochemical, immunocompetent cell functions, and features of the faecal microbiota were analysed after intervention. The oral administration of B. uniformis CECT 7771 reduced body weight gain, liver steatosis and liver cholesterol and triglyceride concentrations and increased small adipocyte numbers in HFD-fed mice. The strain also reduced serum cholesterol, triglyceride, glucose, insulin and leptin levels, and improved oral tolerance to glucose in HFD fed mice. The bacterial strain also reduced dietary fat absorption, as indicated by the reduced number of fat micelles detected in enterocytes. Moreover, B. uniformis CECT 7771 improved immune defence mechanisms, impaired in obesity. HFD-induced obesity led to a decrease in TNF-α production by peritoneal macrophages stimulated with LPS, conversely, the administration of B. uniformis CECT 7771 increased TNF-α production and phagocytosis. Administering this strain also increased TNF-α production by dendritic cells (DCs) in response to LPS stimulation, which was significantly reduced by HFD. B. uniformis CECT 7771 also restored the capacity of DCs to induce a T-cell proliferation response, which was impaired in obese mice. HFD induced marked changes in gut microbiota composition, which were partially restored by the intervention. CONCLUSIONS: Altogether, the findings indicate that administration of B. uniformis CECT 7771 ameliorates HFD-induced metabolic and immune dysfunction associated with intestinal dysbiosis in obese mice.

Gut microbiota dysbiosis is associated with inflammation and bacterial translocation in mice with CCl4-induced fibrosis.

BACKGROUND: Gut is the major source of endogenous bacteria causing infections in advanced cirrhosis. Intestinal barrier dysfunction has been described in cirrhosis and account for an increased bacterial translocation rate. HYPOTHESIS AND AIMS: We hypothesize that microbiota composition may be affected and change along with the induction of experimental cirrhosis, affecting the inflammatory response. ANIMALS AND METHODS: Progressive liver damage was induced in Balb/c mice by weight-controlled oral administration of carbon tetrachloride. Laparotomies were performed at weeks 6, 10, 13 and 16 in a subgroup of treated mice (n = 6/week) and control animals (n = 4/week). Liver tissue specimens, mesenteric lymph nodes, intestinal content and blood were collected at laparotomies. Fibrosis grade, pro-fibrogenic genes expression, gut bacterial composition, bacterial translocation, host's specific butyrate-receptor GPR-43 and serum cytokine levels were measured. RESULTS: Expression of pro-fibrogenic markers was significantly increased compared with control animals and correlated with the accumulated dose of carbon tetrachloride. Bacterial translocation episodes were less frequent in control mice than in treated animals. Gram-positive anaerobic Clostridia spp count was decreased in treated mice compared with control animals and with other gut common bacterial species, altering the aerobic/anaerobic ratio. This fact was associated with a decreased gene expression of GPR43 in neutrophils of treated mice and inversely correlated with TNF-alpha and IL-6 up-regulation in serum of treated mice along the study protocol. This pro-inflammatory scenario favoured blood bacterial translocation in treated animals, showing the highest bacterial translocation rate and aerobic/anaerobic ratio at the same weeks. CONCLUSIONS: Gut microbiota alterations are associated with the development of an inflammatory environment, fibrosis progression and bacterial translocation in carbon tetrachloride-treated mice.

Gut microbiota in obesity and metabolic disorders.

Obesity is a major public health issue as it is causally related to several chronic disorders, including type-2 diabetes, CVD and cancer. Novel research shows that the gut microbiota is involved in obesity and metabolic disorders, revealing that obese animal and human subjects have alterations in the composition of the gut microbiota compared to their lean counterparts. Moreover, transplantation of the microbiota of either obese or lean mice influences body weight in the germ-free recipient mice, suggesting that the gut ecosystem is a relevant target for weight management. Indigenous gut microbes may regulate body weight by influencing the host's metabolic, neuroendocrine and immune functions. The intestinal microbiota, as a whole, provides additional metabolic functions and regulates the host's gene expression, improving the ability to extract and store energy from the diet and contributing to body-weight gain. Imbalances in the gut microbiota and increases in plasma lipopolysaccharide may also act as inflammatory factors related to the development of atherosclerosis, insulin resistance and body-weight gain. In contrast, specific probiotics, prebiotics and related metabolites might exert beneficial effects on lipid and glucose metabolism, the production of satiety peptides and the inflammatory tone related to obesity and associated metabolic disorders. This knowledge is contributing to our understanding of how environmental factors influence obesity and associated diseases, providing new opportunities to design improved dietary intervention strategies to manage these disorders.