Biofilm formation onto starch fibres by Bacillus subtilis governs its successful adaptation to chickpea milk.

Beneficial biofilms may confer effective adaptation to food matrices that assist bacteria in enduring hostile environmental conditions. The matrices, for instance, dietary fibres of various food products, might serve as a natural scaffold for bacterial cells to adhere and grow as biofilms. Here, we report on a unique interaction of Bacillus subtilis cells with the resistant starch fibresof chickpea milk (CPM), herein CPM fibres, along with the production of a reddish-pink pigment. Genetic analysis identified the pigment as pulcherrimin, and also revealed the involvement of Spo0A/SinI pathway in modulating the observed phenotypes. Besides, through successful colonization of the CPM fibres, the wild-type cells of B. subtilis displayed enhanced survivability and resilience to environmental stress, such as heat and in vitro gastrointestinal treatments. In total, we infer that the biofilm formation on CPM fibres is an adaptation response of B. subtilis for strategic survival.

Genetic Architecture Underpinning Yield Components and Seed Mineral-Nutrients in Sesame.

Genetic dissection of yield components and seed mineral-nutrient is crucial for understanding plant physiological and biochemical processes and alleviate nutrient malnutrition. Sesame (Sesamum indicum L.) is an orphan crop that harbors rich allelic repertoire for seed mineral-nutrients. Here, we harness this wide diversity to study the genetic architecture of yield components and seed mineral-nutrients using a core-collection of worldwide genotypes and segregating mapping population. We also tested the association between these traits and the effect of seed nutrients concentration on their bio-accessibility. Wide genetic diversity for yield components and seed mineral-nutrients was found among the core-collection. A high-density linkage map consisting of 19,309 markers was constructed and used for genetic mapping of 84 QTL associated with yield components and 50 QTL for seed minerals. To the best of our knowledge, this is the first report on mineral-nutrients QTL in sesame. Genomic regions with a cluster of overlapping QTL for several morphological and nutritional traits were identified and considered as genomic hotspots. Candidate gene analysis revealed potential functional associations between QTL and corresponding genes, which offers unique opportunities for synchronous improvement of mineral-nutrients. Our findings shed-light on the genetic architecture of yield components, seed mineral-nutrients and their inter- and intra- relationships, which may facilitate future breeding efforts to develop bio-fortified sesame cultivars.

Encapsulation of beneficial probiotic bacteria in extracellular matrix from biofilm-forming Bacillus subtilis.

Probiotics, live microbial supplements, are often incorporated into foods and beverages to provide putative health benefits. To ensure their beneficial effects, these organisms must survive processing and storage of food, its passage through the upper gastrointestinal tract (GIT), and subsequent chemical ingestion processes until they reach their target organ. However, there is considerable loss of viability of probiotic bacteria in the acidic conditions of the stomach and the high bile concentration in the small intestine. Bacillus subtilis, a spore-forming non-pathogenic bacterium, recently has gained interest in its probiotic properties; it can effectively maintain a favorable balance of microflora in the GIT. In addition, B. subtilis produces an extracellular matrix that protects it from stressful environments. We suggested that the extracellular matrix produced by B. subtilis could protect other probiotic bacteria and therefore potentially could be used as a vehicle for delivering viable probiotic cells to humans. Therefore, we developed a novel cultivation system that enables co-culturing of B. subtilis along with probiotic lactic acid bacteria (LAB) by increasing production of the extracellular matrix by B. subtilis cells. Moreover, we showed that B. subtilis improved survivability of LAB during food preparation, storage and ingestion. Therefore, we believe that the results of our study will provide a novel technique of using a natural system for preservation and delivery of probiotics to humans.

Expression of Duodenal Iron Transporter Proteins in Diabetic Patients with and without Iron Deficiency Anemia.

The role of iron transport proteins in the pathogenesis of anemia in patients with diabetes mellitus (T2DM) is still unclear. We investigated the expression of duodenal transporter proteins in diabetic patients with and without iron deficiency anemia (IDA). Methods. Overall, 39 patients were included: 16 with T2DM and IDA (group A), 11 with T2DM without IDA (group B), and 12 controls (group C). Duodenal mucosal expression of divalent metal transporter 1 (DMT1), ferroportin 1 (FPN), hephaestin (HEPH), and transferrin receptor 1 (TfR) was evaluated by Western blotting. Chronic disease activity markers were measured as well. Results. FPN expression was increased in group A compared to group B and controls: 1.17 (0.72-1.46), 0.76 (0.53-1.04), and 0.71 (0.64-0.86), respectively (p = 0.011). TfR levels were over expressed in groups A and B compared to controls: 0.39 (0.26-0.61), 0.36 (0.24-0.43), and 0.18 (0.16-0.24), respectively, (p = 0.004). The three groups did not differ significantly with regard to cellular HEPH and DMT1 expression. The normal CRP and serum ferritin levels, accompanied with normal FPN among diabetic patients without IDA, do not support the association of IDA with chronic inflammatory state. Conclusion. In patients with T2DM and IDA, duodenal iron transport protein expression might be dependent on body iron stores rather than by chronic inflammation or diabetes per se.

Corrigendum to "Expression of Duodenal Iron Transporter Proteins in Diabetic Patients with and without Iron Deficiency Anemia".

[This corrects the article DOI: 10.1155/2018/7494821.].

Enrichment of milk with magnesium provides healthier and safer dairy products.

Biofilms on the surfaces of milk-processing equipment are often a major source of contamination of dairy products. Members of the genus Bacillus appear to be among the most commonly found bacteria in dairy farms and processing plants. Bacillus species may thrive in dairy farm equipment and in dairy products since they can form robust biofilms during growth within milk. We found that fortification of milk with magnesium mitigated biofilm formation by Bacillus species, and thus could notably reduce dairy product spoilage. We also show that the mode of action of Mg2+ ions is specific to inhibition of transcription of genes involved in biofilm formation. Our further findings indicate that in the presence of Mg2+ bacterial cells are hypersensitive to the heat pasteurization applied during milk processing. Additionally, we demonstrated that enrichment of milk with magnesium improved technological properties of milk products such as soft cheeses. Finally, we report that there is a notable increase in the intestinal bioavailability potential of magnesium from supplemented milk compared with that from non-supplemented milk.

α-Linolenic acid (ALA) is an anti-inflammatory agent in inflammatory bowel disease.

Studies suggest that consumption of omega-3 (n-3) polyunsaturated fatty acids (PUFA) plays a protective role in inflammatory bowel disease; however, the use of plant-derived oils rich in α-linolenic acid (ALA) has not been widely investigated. The aims of this study were to test the effects of two different sources of (n-3) PUFA, fish and plant-derived oils, in two animal models of experimental colitis and to determine whether the (n-3) PUFA-enriched diets could ameliorate the inflammatory status. Rats were fed diets rich in corn, fish or sage oil with or without vitamin A supplementation for 3weeks then colitis was induced by adding dextran sodium sulfate to the drinking water or by injecting 2,4,6-trinitrobenzene sulfonic acid. We show that colitic rats fed the sage oil diets had a lower inflammatory response, improved histological repair and had less necrotic damage in the mucosa when compared to the corn and fish oil groups. Colonic damage and myeloperoxidase activity were significantly lower. Colonic mRNA levels of pro-inflammatory genes including interleukin IL-6, cyclooxygenase 2 and tumor necrosis factor α were markedly down-regulated in rats fed fish and sage oils compared to control. These results were supported by experiments in the human colonic epithelial cell line Caco-2, where ALA supplementation was shown to be effective in inhibiting inflammation induced by IL-1ß by down-regulating mRNA levels of pro-inflammatory genes including IL-8, COX2 and inducible nitric oxide synthase. Taken together, these results suggest that plant-derived oil rich in ALA could ameliorate the inflammatory damage in colitis.

Elevated milk ß-carotene and lycopene after carrot and tomato paste supplementation.

OBJECTIVES: To measure concentrations of ß-carotene and lycopene in the breast milk of healthy, well-nourished, lactating women supplemented with fresh carrot or tomato paste. METHODS: Twelve women were given fresh carrot paste and another 14 were given fresh tomato paste once a day for 3 d with a high-lipid-content meal. The women were instructed to avoid any food containing ß-carotene or lycopene, other than the test meal. Milk carotenoid levels were measured before, during, and after the trial. The carrot and tomato meals contained 15 mg of all-trans ß-carotene and 15 mg of all-trans lycopene, respectively. RESULTS: We demonstrated a significant increase in milk ß-carotene and elevated milk lycopene levels after the lipid-rich fresh carrot and tomato meals, respectively. CONCLUSIONS: We suggest that breast milk carotenoid levels reflect the mother's level of intake and can thus be raised by simple nutritional intervention. The results of this study may be relevant to breast-feeding mothers of both preterm and term infants by raising antiinflammatory and antioxidant properties in their milk.

The potential use of chickpeas in development of infant follow-on formula.

BACKGROUND: Undernutrition during childhood is a common disorder in the developing countries, however most research has focussed much on its treatment rather than its prevention. OBJECTIVE: We investigated the potential of using chickpeas in infant follow-on formula production against the requirements of WHO/FAO on complementary foods and EU regulations on follow-on formula. METHODS: Chickpeas were germinated for 72 hours followed by boiling, drying and dehulling in order to minimise associated anti-nutrition factors. Saccharifying enzymes were used to hydrolyse starch to maltose and the resulting flours were analysed for their protein content and amino acid profile. RESULTS: The protein content (percentage) increased from 16.66 ± 0.35 and 20.24 ± 0.50 to 20.00 ± 0.15 and 21.98 ± 0.80 for the processed desi and kabuli cultivar compared to raw chickpeas, respectively (P < 0.05). There was insignificant change (P = 0.05) in amino acid profile following processing and the resulting flour was found to meet the amino acid requirements of WHO/FAO protein reference for 0-24 month's children. CONCLUSION: The designed chickpea based infant follow-on formula meets the WHO/FAO requirements on complementary foods and also the EU regulations on follow-on formula with minimal addition of oils, minerals and vitamins. It uses chickpea as a common source of carbohydrate and protein hence making it more economical and affordable for the developing countries without compromising the nutrition quality.

Needle biopsy for hepatic vitamin A levels in lions (Panthera leo).

Hypovitaminosis A (HA)-related skull malformations resulting in neurologic abnormalities and death have been, and still are, reported in captive lions (Panthera leo) worldwide. Liver vitamin A (VA) concentration is the most reliable indicator of animals' VA status, and its assessment is essential in prevention and treatment of HA in lions. A percutaneous needle liver biopsy using high-performance liquid chromatography ultraviolet retinoid analysis for VA concentration measurement was validated. It was first assessed in vitro using chicken liver. Later, the safety and feasibility of ultrasound-guided percutaneous needle liver biopsy was assessed in living lions. Hepatic VA concentrations in lion liver were measured using the above laboratory method. Mean chicken hepatic VA concentration in needle biopsy (NB) and wedge biopsy (WB) of the same liver lobes were 108.66 and 60.89 microg/g wet tissue, respectively, and were significantly (P = 0.03) correlated (r = 0.74). The calculated linear regression for predicting VA concentration in WB using NB VA for chicken liver was 25.194 + 0.3234x NB (microg/g). Four ultrasound-guided percutaneous needle liver biopsies were obtained from each of the four lions under general anesthesia. Mean hepatic VA concentration was 8.25 microg/g wet tissue (range 1.43-25.29 microg/g). Mean serum VA concentration, measured in these four lions was 1,011.1 nmol/L with a standard deviation of 337.91 nmol/L (range 590.26-1,077.2 nmol/L). The lions recovered uneventfully, and no complications were observed during a 4-yr follow-up period. In conclusion, the percutaneous needle liver biopsy technique is a reliable, practical, safe tool for obtaining liver tissue samples antemortem for assessment of the VA status in lions and can be used in future studies.

Bacterial population and innate immunity-related genes in rat gastrointestinal tract are altered by vitamin A-deficient diet.

Vitamin A and its derivatives have been shown to regulate the growth and differentiation of gastrointestinal epithelial cells; in addition, vitamin A deficiency has been convincingly shown to be associated with increased susceptibility to infection. The gastrointestinal mucosal barrier, which is a component of the innate immune system, is considered the first line of defense, as it provides a barrier between the external environment and the internal milieu. A disturbance in the integrity of the intestinal epithelium is one of the main factors involved in increased incidence of infections during vitamin A deficiency. In this study, the effects of vitamin A deficiency on microbial ecology and the expression of genes related to the intestinal mucosa's innate immunity were examined in a rat model. Using the 16s rDNA method, we demonstrate that a vitamin A-deficient (VAD) diet increases the total amount of bacteria in the gastrointestinal tract and alters the intestinal microflora. Results show a decrease in the relative proportion of Lactobacillus spp. and the simultaneous appearance of Escherichia coli strains. Lack of vitamin A significantly changed mucin (MUC) dynamics, as reflected by the enlarged goblet-cell "cup" area relative to controls; decreased MUC2 mRNA expression in the jejunum, ileum and colon of VAD rats and increased MUC3 mRNA expression in the ileum and colon of these rats. In addition, vitamin A deficiency down-regulated defensin 6 mRNA expression while up-regulating toll-like receptors 2 and 5 mRNA expressions. The current study indicates that vitamin A deficiency interferes with the integrity of the gastrointestinal mucosal barrier.