Mycotoxins are fungal secondary metabolites which are frequently detected in agricultural crops including cereals. They are potent toxins and are important food contaminants which pose a potential risk to consumer health. In addition to fungal mycotoxins, plant-derived mycotoxin metabolites, also called masked mycotoxins, are frequently found in cereals. These masked mycotoxins are sugar-bound metabolites, and their contribution to overall toxicity in humans is less well understood. Hence, this study used a range of in-vitro model systems to predict the intestinal fate of masked mycotoxins in the human gut. The mycotoxins studied included the prevalent Fusarium mycotoxins deoxynivalenol (DON), nivalenol (NIV), T2-toxin (T2), HT2-toxin (HT2) and diacetoxyscripenol (DAS) and their respective glucose-bound metabolites DON-Glc, NIV-Glc, T2-Glc, HT2-Glc, DAS-Glc.

Artificial digestive juices (saliva, gastric juice, duodenal juice) were used to assess the stability of mycotoxins and masked mycotoxins in the small intestine. Human Caco-2 cells were used to study transport, metabolism and toxicity of mycotoxins (1). Fully differentiated Caco-2 cultures were exposed to individual mycotoxins in culture medium in the apical compartment of Transwell filters and absorption and barrier disruption were assessed after 24 hours. Faecal batch cultures using human stool samples and single-strain incubations with important human gut strains were used to assess microbial metabolism of mycotoxins (2). Individual mycotoxins were spiked into microbial cultures and incubated anaerobically for up to 48 hours. Mycotoxins were analysed using LC-MS/MS.

None of the mycotoxins or masked mycotoxins were metabolised by small intestinal juices and absorption studies showed that free mycotoxins are readily absorbed and can disrupt intestinal barrier function. In contrast, masked mycotoxins are not absorbed though the epithelial monolayer intact and can hence be transported to the large intestine. Upon contact with microbiota, masked mycotoxins were readily hydrolysed and free mycotoxins released into the culture supernatant. Marked differences were observed in the hydrolysis rates between different donor samples as well as between different masked mycotoxins treated with the same donor sample. Microbial de-acetylation was observed for several mycotoxin in all donor samples, whereas de-epoxydation was rare. These results suggest that human gut microbiota are not efficient in detoxifying dietary mycotoxins. The rapid release of free mycotoxins from masked forms by human gut microbiota demonstrates that masked mycotoxins will contribute towards overall toxicity in humans and need to be included in risk assessments.

This work has been supported by the Scottish Government Rural and Environment Science and Analytical Services Division (RESAS) and a grant from Food Standards Agency (FSA).



1.	Gratz SW, Dinesh R, Yoshinari T, Holtrop G, Richardson AJ, Duncan G, MacDonald S, Lloyd A, T

Gut microbiota metabolise food-derived mycotoxins

Yakult

There is a lot of agricultural waste that still needs to be used for efficient products, such as banana flower or Ontong and banana leaves. Based on the studies, banana flowers and banana leaves (Musa balbisiana), especially the type of kluthuk, contain high nutrients, such as fiber and starch and other bioactives. However, its use for health products is still lacking. This study aims to process or formulate and utilize banana flowers and banana leaves of the kluthuk type to become a functional food product in the form of fermented drinks. In addition, it is also to determine the levels of antioxidants (especially vitamin C) in fermented products in vitro.
There are 3 variations of the formulation, Banana Flower: Banana Leaf: CO2 free water. Sample 1 (1: 0.5: 0.5), sample 2 (2: 1: 1) and sample 3 (3: 2: 2). Then, all product samples were inoculated with Lactobacillus paracasei for 14 days under anaerobic conditions. Sample variation was carried out to determine the significance of the average antioxidant content in it. The next step was to test the analysis of vitamin C from 3 samples using the Iodometric Titration Method, to determine the amount of Vitamin C (mg/100g) and the antioxidant activity with 2.2-diphenyl-1-picrylhydrazyl (DPPH) with triplo repeat per sample.
The amount of vitamin C obtained at S1 was 60.40±1.00 mg/100g with 15.55±0.05 % antioxidant activity, S2 was 55.95±0.05 mg/100g with 10.60±0.30 % antioxidant activity and S3 was 77.92±0.02 mg/100g with 22.10±0.10 % antioxidant activity. The fermented drink sample formulation containing the highest vitamin C was S3. There was a significant difference (P<0.05) which determined the vitamin C levels between the sample formulations. The average ash content of the three samples was 2.40±0.40 % and the moisture content was 35.60±5.00 %. The average vitamin C level in the three fermented drink samples was 64.75±11.61 mg/100g. S3 showed the best activity, namely antioxidant activity against 2.2-diphenyl-1-picrylhydrazyl (DPPH) of 22.10±0.10 %.
The combination of kluthuk banana flowers and leaves has a great potential to be developed into health functional food. By looking at the content of vitamin C and antioxidants in the fermented products, it can be an effort to diversify local food and also become a good drink during the pandemic, because antioxidants have the ability to increase immunity and anti-inflammation. This product also contains prebiotics and probiotics which are good for the gut microbiome which play a good role in the immune system. Therefore, these product can be a potential Anti SARS-CoV-2, which SARS-Cov-2 itself attacks immunity and causes inflammation in the body. Animal studies are needed to find out the interactions between substances before they are tested in humans.

Acknowledgements: We thank to all of contributors for their outstanding help in formatting the abstract, especially Fahrul Nurkolis who has fully assisted in laboratory tests. The authors’ responsibilit

Probiotic drink from the fermentation of banana flower (Ontong) with addition of kluthuk banana leaves (Musa balbisiana)

Quorn Foods

Introduction: Gut microbiota modulation by nutritional procedures has become a feasible therapeutic target in the management of obesity and non-alcoholic fatty liver disease (NAFLD) (1). Recently, Akkermansia muciniphila has been considered as a potential probiotic due to its protective effect in obesity development (2). Besides, the flavonoid quercetin has shown an anti-inflammatory, antioxidant and prebiotic capacity, highlighting a great therapeutic potential in NAFLD (3). This study aims to evaluate the effect of gut microbiota modulation by the feasible symbiotic quercetin and A. muciniphila combined with nutritional intervention in the development of early obesity-related NAFLD in an in vivo model.

Methods: 21-days old Wistar rats were fed with a high-fat diet (HFD) or a control (C) diet for 6 weeks. Afterwards, all animals were fed with a C diet supplemented or not with quercetin and A. muciniphila (administered daily by oral gavage) for 3 weeks. Faecal samples were analyzed by qPCR and sequenced by Illumina MiSeq System to determine gut microbiota composition. Statistical significance was determined by nonparametric Kruskal-Wallis test followed by Mann-Whitney U test (p<0.05).

Results: After 6 weeks, HFD intake triggered a lower concentration of the total bacteria DNA in comparison to C, but increased gut microbiota diversity calculated by the Shannon diversity index. Metagenomic analysis demonstrated an HFD-related gut dysbiosis, showing a significant increase of Firmicutes, Proteobacteria and Actinobacteria phyla, while, Bacteroidetes and Cyanobacteria showed an opposite pattern. At genus level, HFD increased the relative abundance of Adlercreutzia, Allobaculum, Bifidobacterium, Bilophila, Blautia, Clostridium, Enterococcus, Lactobacillus, Oscillospira, Rothia and Streptococcus genera. In contrast, Coprobacillus, Coprococcus, Phascolartobacterium, Prevotella, Roseburia, Sutturella and Turicibacter were reduced. Principal coordinates analysis (PCoA) revealed that the bacterial communities of HFD-fed rats clustered separately from those of C group. After 3 weeks, the nutritional intervention together with quercetin and A. muciniphila supplementation reversed the gut dysbiosis associated to HFD, tending to increase total bacteria counts and decrease Shannon diversity index. Moreover, a notable increase in the abundance of Cyanobacteria phylum, as well as a decrease in Actinobacteria phylum was detected. Additionally, several differences at genus level were observed in the symbiotic supplemented groups that were not detected with the administration of any of the treatments alone. The symbiotic reduced the detection of Lactococcus and Lactobacillus genera, whereas Oscillospira was highly abundant in both supplemented groups. To determine the effect of quercetin and A. muciniphila administration on bacterial communities a PCoA was conducted, showing that the individuals receiving the symbiotic tended to cluster together and separated from t

A dietary intervention with Akkermansia muciniphila and quercetin supplementation reshapes gut microbiota composition in an in vivo model of early obesity related non-alcoholic fatty liver disease

Clasado

Nutrium

Introduction: Obesity has emerged as one of the major global public health concerns, associated with a significant morbidity and mortality. Obesity clinical management demands increasing efforts for new therapeutic targets and innovative approaches. The link between gut microbiota and obesity has been thoroughly demonstrated, pointing gut microbiota manipulation as a promising tool to address this issue(1). Bariatric surgery is a reliable resource to deal with severe cases when other approaches have failed. However, it has been stated that it may have a profound impact on gut microbiota which is still poorly understood(2). We aim to characterise the changes in fecal microbiome and metabolome associated with bariatric surgery on morbid obese patients.
Methods: Ten patients (20-60 years old) diagnosed with obesity not treated with antibiotics 1 month before the study were recruited by Endocrinology Service of Complejo Asistencial Universitario de León. Fecal samples were collected before and after bariatric surgery and processed for metabolomics and metagenomic analysis. Microbiota composition was identified by 16S rRNA Illumina MiSeq sequencing. Polar metabolites were quantified by liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry was employed to determine short chain fatty acids (SCFAs) concentration. Kruskal-Wallis followed by Wilcoxon test for paired samples allowed identification of varying taxa. Univariant analysis according to fold change and significant Wilcoxon test, and multivariate analysis based on PLS-DA models were employed to estimate differences associated to surgery.
Results: Bacterial communities were modified by bariatric surgery, tending to cluster separately on β-diversity plot based on UniFrac distance, while α-diversity remained unaffected. More remarkable differences were detected at phylum and genus levels. Relative abundance of Tenericutes, Firmicutes, Saccharibacteria (phyla), Coprococcus, Lactococcus and Lachnospira (genera) decreased after surgery while increases in Butyricimonas, Parabacteroides and Slackia were detected. Fecal metabolome composition was deeply modified by bariatric surgery. All predominant SCFAs (acetate, propionate and butyrate) experimented a significant decrease upon surgery (-52.2±15.40%, -46.2±11.92% and -63.5±9.26%, respectively). Moreover, metabolomic analysis highlighted the contribution of a subset of metabolites with a great performance on classifying samples in pre and post-operated. Among these compounds, intermediates of tricarboxylic acids cycle like citrate and malate, amino acids like aminolevulinate and histidine and others like taurine, glyoxylate or choline (all experimenting decreases) highlighted as the ones with larger differences following bariatric surgery, being considered hallmarks of the procedure and a potential fingerprint of weight loss in obese patients.
Conclusion: Bariatric surgery reshapes gut microbiota composition and produces signific

Metagenomic and metabolomic shift on morbid obese patients undergoing bariatric surgery

Cambridge University Press

Introduction: 
Banana tree (Musa parasidica) could be environmental waste. The tree can be extracted to become starch, can be used as innovative product that contain high food fiber, and can be potential probiotics when adding pineapple pulp (Ananas comosus). 
This study aims to process or formulate and utilize banana tree starch with the addition of pineapple pulp into a functional food product in the form of probiotic drinks. In addition, in vitro levels of antioxidants (especially vitamin C) and total dietary fiber were also carried out.

Methods:
There are 3 variations of the formulation, Banana Tree Starch: Pineapple: CO2 free water. Sample 1 (1: 0.5: 0.5), sample 2 (2: 1: 1) and sample 3 (3: 2: 2). Then, all product samples were inoculated with Lactobacillus paracasei for 60 hours and incubated in anaerobic conditions with a temperature of 30-32°C. Sample variation was carried out to determine the significance of the average antioxidant content in it.
The next step was to analyze the vitamin C from 3 variants of beverage samples by using the Iodometric Titration Method that determined the amount of vitamin C (mg/100g), and also measured the antioxidant activity by using 2.2-diphenyl-1-picrylhydrazyl (DPPH). Determination of dietary fiber content using the AOAC method. All with triplo repeat per sample.

Results:
The amount of vitamin C obtained in S1 was 80.45±0.05 mg/100g with 44.95±0.99 % antioxidant activity and 11.01±0.01 % dietary fiber. The S2 vitamin C was 65.75±0.05 mg/100g with antioxidant activity 30.60±0.30 % and 8.90±0.10 % dietary fiber, and the last S3 was 47.02±0.02 mg/100g with 32.10±0.20 % antioxidant activity and 7.89±0.10 % dietary fiber. The average vitamin C level in the three fermented drink samples was 64.40±16.75 mg/100g, while the highest concentration of vitamin C was S1. There was a significant difference (p < 0.05) of the vitamin C levels amongst the three sample formulations. The average ash content of the three samples was 4.77±0.07 % and the moisture content was 37.72±7.72 %. The S1 sample showed the best antioxidant activity against 2.2-diphenyl-1-picrylhydrazyl (DPPH) which was 44.95±0.99 % and also had highest level of dietary fiber.

Conclusion: 
The formulation of banana tree starch combination with pineapple pulp has greatly potential to be developed into a health functional food. By looking at the content of vitamin C and total dietary fiber and antioxidant activity in the fermented products, this can be an effort to diversify local food and can be an alternative to antidiabetic drinks and other infectious diseases.

Functional drink formulation from the fermentation of banana tree (Musa parasidica) rich in antioxidant and food fiber

Background: Protein supplements have become a widespread practice amongst athletes and gym users worldwide. However, the use of protein supplements among female gym users in Riyadh, Saudi Arabia has not yet been investigated. 
Objectives: The aim of this study was to investigate the prevalence of protein supplement usage among female gym users and their attitudes towards protein supplementation in Riyadh. 
Methods: In this cross-sectional study, 502 female gym users aged ≥18 years completed a structured, self-administered online questionnaire on the usage of protein supplementation. Socio-demographic information, anthropometry, health status, exercise frequency, attitudes and knowledge regarding protein supplements were obtained.
Results: The overall prevalence rate of protein supplement use among the study respondents was 36.3%. There were significant associations between the use of protein supplements and educational level (p< 0.05), the duration of being a gym member (p< 0.05) and frequency of gym visits per week (p< 0.05). Whey protein was the predominantly used supplement (57.8%) and the reason attributed to the use of protein supplement was muscle building (53.9%). The main sources of information regarding protein supplement use included the internet (30.2%) followed by gym coaches (25.3%).
Conclusions: Our findings demonstrated that a considerable number of females exercising in gyms use protein supplements and most rely on unreliable sources for decision-making of protein supplements use. Nutritional education and consultations regarding protein supplement use are warranted.

Prevalence and attitudes towards using protein supplements among female gym users: An only survey

Introduction:
Mango (Mangifera indica) is a tropical fruit that grows abundantly in Indonesia and has a great potential to be developed into a high-value product rather than being eaten raw, namely as a functional drink. There are several scientific studies that show mango (1) to be a fruit rich in dietary fiber, vitamin C, vitamin E, β-carotene, and polyphenols of the types of quercetin and kaempferol that have the potential to improve lipid profiles and lower blood glucose. In addition, spinach (Amaranthus) is a vegetable that has a nutritional content similar to mangoes and even more, such as high polyphenols. However, there is still little use of it into fermented beverage products. This study aims to process or formulate and utilize mango pulp and spinach flour to become functional food products in the form of fermented drinks. In addition, it is also to determine the levels of polyphenols (especially quercetin and kaempferol) and the total dietary fiber in these fermented products in vitro.
Methods:
The formulation was carried out on 3 samples with different amounts (mango pulp: spinach flour: water) for 3 samples each S1 (900: 100: 0), S2 (850: 150: 150) and S3 (800 : 200: 200). Each sample was inoculated with Lactobacillus paracasei for 60 hours and incubated in anaerobic conditions with a temperature of 30-32°C. Analysis of polyphenols (quercetin and kaempferol) using the High Performance Liquid Chromatography (HPLC) technique at a wavelength of 415 nm with triplo repeat per sample. Determination of dietary fiber content using the AOAC method. The statistical test is to determine the difference between mango and water formulations on the polyphenols content of fermented products used One Way Anova.
Results:
It was found that the amount of quercetin and kaempferol in S1 was 55.89±0.01 µg / g and 59.98±0.02 µg / g, respectively, for S2 were 40.66±0.02 µg / g and 50.42±0.04 µg / g, and in S3 51.90±0.05 µg / g and 48.88±0.08µg / g. There was a significant difference (P<0.05) which determined the polyphenols content between the sample formulations. The content of dietary fiber at S1 was 11.70±1.00 %, S2 was 9.80±0.06 % and at S3 was 10.01±0.01 %. The average ash content of the three samples was 3.84±0.04 % and water content was 42.01±2.01 %.
Conclusion:
The highest polyphenols content was found in S1 samples with the highest levels of dietary fiber as well. Therefore, mango and spinach have great potential to be developed into fermented functional foods or probiotic drinks. The content of polyphenols (quercetin and kaempferol) in these fermented products can be a great alternative to ready-to-drink drinks, because these types of polyphenols (quercetin and kaempferol) have the ability to improve lipid profiles and stabilize blood sugar fluctuations so that they can be anti-diabetics (2).
Acknowledgements:
We thank to all of contributors for their outstanding help in formatting the abstract. The authors’ responsibilities were as follows—all aut

Probiotic drink from fermented mango (Mangifera indica) with addition of spinach flour (Amaranthus) high in polyphenols and food fiber

Introduction: Irritable bowel syndrome (IBS) is a functional bowel disease accompanied by symptoms such as chronic abdominal pain, bloating, gas and defecation disorders triggered by some dietary compounds and foods. A gluten-free diet and recently approved diet called a low-FODMAP diet, which affects dietary soluble and insoluble fibre intake are applied to relieve these symptoms⁽¹⁾. These diets affect gut microbiomes that include beneficial microorganisms for human health⁽²⁾. This study was aimed to evaluate different medical nutrition therapies on dietary fibre intake, which may cause dysbiosis in the gut.

Methods: In this study, forty-five patients aged 19-64 diagnosed with constipation-predominant IBS (IBS-C) according to Rome-IV criteria were randomized into three groups of 15 patients each. Each group adopted either a gluten-free diet, low-FODMAP (fermentable oligo-, di-, monosaccharides and polyols) diet or traditional constipation diet for six days. Participants were asked to keep a food diary using 24-hour food record for each day. At the end of the study, the food diaries were evaluated in total dietary soluble and insoluble fibre intake using Nutrition Information System software (BeBiS).

Results: Participants mean age was 37.6±11.2 year, and body mass index 25.6±3.2 kg/m2. Mean total dietary soluble fibre intake of traditional constipation diet group was 12.8±4.4 g, gluten-free diet group was 11.6±3.1 g and low-FODMAP diet group was 9.0±4.0 g. The difference in total dietary soluble fibre intake between groups was statistically significant (p = 0.033). Mean total dietary insoluble intake of traditional constipation diet group was 22.9±6.6 g, gluten-free diet group was 23.2±5.3 g, and low-FODMAP diet group was 16.9±6.9 g. Total dietary insoluble intake was the lowest for low-FODMAP diet groups and found statistically significant (p=0.024).

Conclusion: Gut microbiome needs to be supported by diet to increase its number and maintain host health. Dietary fibre, especially soluble fibre, is one of the primary energy sources of the microbiome. Even though low-FODMAP and gluten-free diets are effective to relieve the symptoms of IBS-C, they are restricted from dietary fibre. Compared to these two diets, a traditional constipation diet includes more fibre however it is not enough for a healthy diet. To support patients' dietary intervention with enough soluble fibre may be helpful for the gut microbiome.

The dietary fiber intake of patients with constipation-predominant irritable bowel syndrome who applied different medical nutrition therapies

Frailty and malnutrition are widespread syndromes in older adults, and are both linked to increased risk for dependency, hospitalisation and mortality (1). Evidence suggests that these conditions share common causal factors and physiological pathways, such as body composition changes and loss of tissue mass (2). The objectives of this study were to investigate the prevalence of, and association between, frailty and malnutrition, in addition to factors associated with each condition, in a cohort of Irish adults aged ≥65 years.
A total of 142 community-dwelling older volunteers (n = 81 female, n = 61 male, 74.1 ± 6.80 years) were recruited to participate through advertisement in community settings in Cork, Ireland (March-August, 2019). Community-based screening sessions were held for those eligible, where frailty, nutritional status and dietary variables were measured. Frailty was classified according to the Frailty Phenotype, described as ≥3 (or 1-2 for pre-frailty) of the following features; weakness, exhaustion, low activity, slowness and weight loss. Nutritional status was assessed using the Mini Nutritional Assessment – Short Form (MNA-SF). Supplement use and intake of protein (%E) and alcohol (g) were recorded using a validated Food Frequency Questionnaire (FFQ). Pearson’s chi-squared test and logistic regression analysis were performed to investigate the association between frailty, malnutrition and demographic and dietary variables. Ethical approval was obtained from C.I.T. Research Ethics Committee, Cork, Ireland. 
The prevalence of pre-frailty and frailty in this cohort were 34.5% (n = 49) and 12.0% (n = 17), respectively. Only one participant was malnourished, thus, those who were malnourished and at risk of malnutrition were grouped together for analysis, resulting in a prevalence of 19.7% (n = 28). No significant differences were observed in the prevalence of frailty or malnutrition risk between genders (p >0.05). Of the 28 participants that were malnourished/at risk, 21 were also pre-frail or frail, while only 7 were non-frail (p <0.001). Use of dietary supplements decreased the risk for malnutrition [odds ratio (OR) (95% CI): 0.19 (0.04-0.68), p = 0.021], while being pre-frail or frail (compared to non-frail) increased the risk for malnutrition [OR: 5.06 (1.82-15.56), p = 0.003]. Increasing age, being malnourished/at risk (compared to well-nourished) and having a low protein intake were associated with an increased frailty risk [OR: 1.16 (1.10-1.23), 5.83 (2.36-14.95) and 2.93 (1.16-7.62), respectively, all p <0.05]. 
Our study revealed a frailty prevalence of 12.0% in this cohort, while 19.7% were malnourished or at risk of malnutrition. Despite a divergence in risk factors, a significant overlap exists between frailty and malnutrition. The direction of this association remains unclear, however, and warrants further exploration. Nonetheless, nutritional considerations are important for the development of strategies to reduce/prevent

Frailty and malnutrition in a cohort of Irish community-dwelling older adults.

Prolonged oxidative stress, induced by cellular metabolic processes and/ or environmental factors, can contribute to numerous chronic non-communicable diseases including neurodegenerative diseases, cancer, diabetes, liver injury, rheumatoid arthritis, respiratory disease, and accelerated ageing (1). This study investigated the cellular antioxidant activity of six blue whiting soluble protein hydrolysates (BWSPH, BW-SPH-A - BW-SPH-F), produced using a proprietary process, using RAW264.7 murine macrophage cells in culture. The antioxidant activity of the BWSPHs was determined via investigating their ability to scavenge the 1-diphenyl-2-picrylhydrazyl (DPPH) radical, as well as, to inhibit the production of reactive oxygen species (ROS) and to promote the expression of the endogenous antioxidant glutathione (GSH) and catalase (CAT) enzyme activity in oxidant-treated RAW264.7 cells. Statistical analysis was determined using ANOVA followed by Dunnett’s test (Prism 5.0, GraphPad Inc. San Diego, CA, USA). Results are expressed as the mean of at least three independent experiments ± SE. 
Hydrolysate BW-SPH-A exhibited superior DPPH radical scavenging activity compared to the other five hydrolysates, inhibiting DPPH by 50% at a concentration of 2.10 ± 0.12 mg/ mL. BW-SPH-A (0.5% w/v) was the only hydrolysate to significantly increase GSH and CAT levels in tert-butyl (tBOOH)- and hydrogen peroxide (H2O2)- challenged RAW264.7 cells, respectively, compared with the treated controls (p<0.05). Exposure of RAW264.7 cells to BW-SPH-A (0.5% w/v) for 24 h prior to treatment with H2O2 also significantly reduced (p<0.05) the generation of ROS compared to cells treated with H2O2 alone. 
The harsh conditions and oxidative nature of the gastrointestinal (GI) tract can alter the peptide profiles of the various BWSPHs and ultimately influence their antioxidant capacity. Simulated gastrointestinal digestion (SGID) of BWSPHs mimics this digestion process, thereby enhancing the physiological relevance of in vitro studies. For BWSPHs to have use as antioxidant agents, their digests must be able to transport across the intestinal barrier, eventually reaching the target cells and effectively stimulate endogenous antioxidant defence systems for the elimination of free radicals.  
The digest of hydrolysate BW-SPH-A (i.e., BW-SPH-A-GI) also increased GSH concentration in tBOOH-treated RAW264.7 cells and reduced ROS production in RAW264.7 cells challenged with H2O2 compared to treated controls (p<0.05). Additionally, the digest of BW-SPH-E (i.e., BW-SPH-E-GI) significantly increased CAT activity in H2O2-challenged RAW264.7 cells compared to the H2O2 control (p<0.05). It is possible that SGID of BW-SPH-E released antioxidant peptide(s) which were responsible for promoting CAT activity in oxidatively-stressed cells.
In conclusion, it is possible that hydrolysate BW-SPH-A, which exhibited antioxidant activity in vitro, may also protect against oxidative stress in vivo and thereby, ma

Antioxidant activity of blue whiting protein hydrolysates- an in vitro study

Introduction: Bariatric surgery can provide weight loss through not only reducing dietary intake but also modifying metabolic control, hunger-satiety mechanisms and gut microbiome (1-4). This study investigated the effects of bariatric surgery on dietary intake, gut microbiome and metabolic markers. 
Methods: The study included 15 morbid obese who had bariatric surgery after baseline interview (Study Group), 8 morbid obese (Control group-1) and 11 non-obese individuals (Control group-2). All participants were followed-up for a 6-month period with face-to-face interviews at baseline (M0), at the end of 3 (M3) and 6 months (M6). Nutritional assessment including dietary intake and anthropometry was done, blood samples were taken for biochemical analysis, and faecal samples were collected for gut microbiome analysis at M0, M3 and M6. Microbiota analysis was performed using QIIME pipeline. Kruskal-Wallis, Wilcoxon and Spearman’s rank correlation tests were performed with SPSS 23.0. 
Results: Participants in the study group lost about 33 kg of body weight in 6 months, in addition to declines in other anthropometric measurements (p<0.05, for each). Dietary energy and nutrient intakes decreased immediately after surgery. Serum glucose, total cholesterol, LDL-cholesterol and hs-CRP levels decreased by surgery (p<0.05, for each). No changes in anthropometrical measurements, dietary intake or biochemical parameters were observed in any of control groups during follow-up period (p>0.05, for each). Morbidly obese participants had higher levels of Firmicutes and lower levels of Bacteroidetes than non-obese subjects at the baseline. By the surgery, the abundances of phylum Bacteroidetes increased from 27.9% to 46.0% (p=0.02) and phylum Firmicutes decreased from 50.3% to 41.7% (p>0.05), leading a significant decrease in Firmicutes/Bacteroidetes ratio (p=0.01) from M0 to M6. At genus level, abundance of Lactobacillus and Bifidobacterium decreased while Akkermansia increased at M6 in study group (p<0.01, for each). In terms of diversity, morbidly obese participants had a distinct profile according to ß-diversity indices at M0. However, it changed towards the profile of non-obese participants at M3, and became similar with them at M6 (p>0.05) in study group. Besides, α-diversity indices were lower in morbidly obese subjects at M0, but became similar to levels in non-obese controls at M6. No correlation between the alterations in biochemical parameters and relative abundances of Bacteroidetes and Firmicutes phyla was observed, however, decline in serum total and LDL cholesterol levels were negatively correlated with decrease in the relative abundance of Proteobacteria (r=-0.718, p=0.013). Furthermore, decrease in the abundance of Firmicutes positively correlated with the changes in both total fat and monounsaturated fatty acid intakes (r= 0.575, p=0.025).
Conclusion: This study showed the substantial effects of obesity and bariatric surgery on gut microbiome profile

Potential associations between alterations in gut microbiome and obesity-related traits after the bariatric surgery

The glycaemic index (GI) of carbohydrate foods is a useful tool for predicting postprandial glucose and insulin responses (1). Results from human studies suggest an inverse relationship between GI of ingested food and postprandial appetite responses (2). However, evidence on the effects of GI on subjective and objective satiety responses is inconclusive (3). Therefore, the aim of this study was to assess the impact of GI on postprandial appetite response in humans.
We conducted a randomised controlled crossover trial on 13 adults presenting with normoglycaemia, normal BMI and mean age of 28.7 ± 6.6 years. Participants were randomly allocated to consume a high GI meal (HGI) consisting of mashed potatoes, and low GI (LGI) meal of canned whole chickpeas, each providing 50 g total available carbohydrates, after an overnight fast. Postprandial subjective satiety and appetite responses were captured using visual analogue scale (VAS) in 3 domains: hunger, fullness, and prospective food intake. Plasma ghrelin values were also measured by immunoassay (intra-assay variations < 15%), in venous blood samples collected over 3 hours following breakfast intake. Postprandial total area under the curves (AUCs) were calculated by the trapezoidal method, and data were analysed using t-test with statistical significance set at p < 0.05. Data are presented as mean ± SD.
There were great variations observed in individual responses after intake of both interventions in all outcomes illustrated above. Postprandial hunger responses were significantly lower after ingestion of LGI meal (91.2 ± 37.1 mm × h) in comparison to HGI meal (113.5 ± 26 mm × h) (p = 0.035); and postprandial fullness responses were significantly higher as demonstrated by AUC values over the 3 hour period following breakfast intake (107.3 ± 37.1 mm × h) and (80.2 ± 24.7 mm × h) (p = 0.012) respectively. However, there was no significant difference in prospective food intake following different GI meals (104.6 ± 41.9 mm × h) and (126.2 ± 26.3 mm × h), respectively. In addition, there were no differences observed in postprandial ghrelin response (p > 0.05).
The results suggest that a low meal GI positively affects postprandial satiety response. However, the impact on appetite hormone ghrelin is unclear possibly due to high individual variances in hormonal responses.

Influence of glycaemic index on subjective appetite responses in healthy adults

Objectives 

Dietary fibre intake levels in UK adults are not currently meeting the recommended 30g per day guidance(1). Individuals are encouraged to increase their fibre intake, however, this may result in an increase in digestive symptoms in some people(2). We aimed to explore current experiences of digestive symptoms in the UK adult population and determine associations with lifestyle, diet and  demographic factors. 

Methods 

An online self-administered survey was developed using an online tool (Online Surveys), and participants were recruited via social media sites including Facebook, Twitter, LinkedIn, Reddit, forums and via word of mouth. Statistical analysis of binary or categorical data (chi squared tests) was carried out using SPSS.  

Results 

The survey was completed by 238 individuals aged 18-82 years (median = 34.5); 77% female, 22% male and 1% other. At least 1 digestive symptom was reported by 81% of respondents in the previous 4 weeks. For 91% of respondents this was their normal experience of digestive symptoms. The most commonly reported symptoms were flatulence (67%), abdominal bloating (58%) and borborygmi (54%). The number of different symptoms experienced by females (median = 5) in the previous 4 weeks was significantly greater than males (median = 1) (p<0.001). Significantly more females (32%) rated symptoms as severe compared to males (13%) (p<0.05). Those who ate ≥5 portions of fruit and vegetables per day rated symptom bother as lower (median = 3 (out of 10)) than those who reported eating <5 portions (median = 5) (p<0.05). The lifestyle factors most commonly reported as perceived causes of digestive symptoms were work stress (38%), personal life stress (37%) and eating too fast (32%). When asked whether digestive symptoms ever negatively impact their life 47% of respondents answered yes. Energy levels (13%) and diet (12%) were most commonly identified as often being negatively impacted by digestive symptoms followed by body image (11%), daily activities, leisure activities and social life (all 8%). Significantly more females reported that digestive symptoms had a negative impact on social life (n=20), body image (n=27) and energy levels (n=30) than males (n=1, 2 and 2 respectively) (p<0.05). 

Conclusions 

Digestive symptoms were common among the respondents of this survey, and the results in relation to the female experience of digestive symptoms support previous research findings(3). The results highlight the multifactorial influences that contribute to digestive symptoms and the impacts they can have on those who experience them.

Digestive symptoms, diet and demographics in UK adults

Efficacy of nutrition interventions in ameliorating malnutrition among children: a systematic review
E. E. Payin1 and P. Jacob2
1.	Department of Food and Nutrition Education, Faculty of Home Economics Education
University of Education, P. O. Box 25, Winneba, Ghana and 2. Atlantic International University, Pioneer Plaza, 900 Fort Street Mall 905, Honolulu, HI 96813, Hawaii, USA

Introduction
Malnutrition is a state of deficiencies, excesses, or imbalances in a person’s intake of energy and or nutrients(1). Undernutrition, micronutrient-related malnutrition and overweight, obesity and diet-related non-communicable diseases are the three broad groups of conditions that malnutrition encompasses(2). Around 45% of deaths among children under age 5 years are linked to Undernutrition(1). In 2019, globally, about 144 million children less than 5 years of age suffered from stunting, 47 million children under age 5 were wasted of which 14.3 were severely wasted and 38.3 million were overweight (3). The goal of nutrition intervention is to resolve or improve a nutrition diagnosis or nutrition problem by the provision of advice, counselling, education, or delivery of a food component of a specific diet or meal plan tailored to a patient’s needs(4). Thus the objective of this systematic review was to evaluate the efficacy of the nutrition interventions administered for the treatment of malnutrition among children under the age of 5.

Methods
Design
Systematic review
Data Sources
Maternal and Child Nutrition Journal, BMJ Global Health, The American Journal of Clinical Nutrition, Public Health Nutrition and the Journal of Nutrition studies were identified from Scopus and Schimago database. 

Eligibility Criteria for Selecting Studies
Randomised controlled trials, cohorts and case studies published between January 2013 and September 2020 and that focused on treatment and management of malnutrition among children under the age of 5 were eligible for inclusion. Authors independently extracted the data and assessed the risk of bias of 10 included randomised control trials.

Result
The 13 included studies contained a sample of 45,781 under 5 children from Low and Middle-Income Countries diagnosed with severe acute, moderate acute, global acute, severe wasting, moderate wasting and stunting. Food deliveries, such as therapeutic, supplementary and augmented foods, nutrition counselling and education were the main interventions used. Locally developed therapeutic and supplementary ready-to-use foods were found to have similar high recovery rates from malnutrition as the standardised ones. Interventions (6/8) that combined both food delivery and either nutrition counselling or education yielded higher recovery from malnutrition. Milk, egg, and multiple micronutrient supplementations improved linear growth of +0.23 (95% CI: 0.18, 0.29; P < 0.05) of stunted children. About 90.3% of SAM children treated with RUTF and high dose of vitamin D3 gained ˃ 15% of the baseline body

Efficacy of nutrition interventions in ameliorating malnutrition among children: a systematic review

Introduction: Mediterranean diet has been recommended for type 2 diabetes treatment(1). The 
impact of diet in shaping the gut microbiota is well known, particularly Mediterranean diet(2–
4). However, the link between Mediterranean diet and diabetes outcomes improvement is not 
completely clear. The aim of this clinical trial was to study the role of microbiota modulation 
by a Mediterranean diet intervention on the clinical outcomes in a diabetic population. 
Material and Methods: In this 12-week single-arm pilot study, nine participants received 
individual nutritional counselling sessions promoting Mediterranean diet. Gut microbiota was 
analysed by next-generation sequencing at baseline, 4 weeks, and 12 weeks after the 
intervention. Biochemical parameters, body composition and blood pressure were assessed at 
the same time-points (baseline, 4 weeks, and 12 weeks). Mediterranean diet adherence was 
assessed by MEDAS score at baseline and after 12 weeks. Statistical analysis was performed 
using One-way ANOVA followed by Bonferroni test for parametric variables and Friedman 
test followed by repeated Wilcoxon tests for non-parametric variables. Effect sizes (Cohen d 
test) are also presented to decide whether a clinically relevant effect is found (|d| > 0.50 indicates 
a medium effect size and |d| > 0.80 a large effect size)(5). This study was approved by the Ethics 
Committee of Administração Regional de Saúde de Lisboa e Vale do Tejo (Ref. 016/CES/ 
INV/2019) and Faculdade de Ciências Médicas|NOVA Medical School, Universidade NOVA 
de Lisboa (Ref. 55/2018/CEFCM) and was registered at clinicaltrials.gov as NCT04403217.
Results: Data are presented as mean±SD. Adherence to the Mediterranean diet (MEDAS score) 
increased from 8.11±2.26 at baseline to 10.8±1.25 after the intervention (p < 0.05; Cohen d= 
1.58). Changes in microbial richness was observed right after 4 weeks of the intervention 
(73.14±39.91 to 100±41.5, p= 0.205; Cohen d= 0.66), being negatively correlated with fasting 
glucose levels (r= -0.634; p < 0.05) and HOMA-IR (rs= -0.464; p < 0.05). On the other hand, 
HbA1c decreased from 7.53±1.07% to 6.86±0.85% (p < 0.05; Cohen d= -0.70) and HOMA-IR 
decreased from 3.79±2.98 to 2.76±2.05 at the end of study (12 weeks) (p < 0.05; Cohen d= -
0.41). Alkaline phosphatase activity was assessed in faecal samples and was negatively 
correlated with HbA1c (rs= -0.584; p < 0.05) and positively correlated with microbial diversity 
(rs= 0.608; p < 0.05).
Conclusions: This study reinforce that increasing adherence to the Mediterranean diet resulted
in a better glycaemic control in subjects with type 2 diabetes. Gut microbial richness changes seemed relevant in mediating the metabolic impact of therapeutic interventions and may 
constitute new target for the treatment of type 2 diabetes. Nonetheless, the number of 
participants that enrolled in this pilot study was small (9 participants), thus studies with a larger 
sample size are needed to c

Gut microbial richness as an earlier biomarker of Mediterranean diet intervention in type 2 diabetes metabolic control

Food sources and inulin consumption in school-aged children.

Introduction
Dietary fibre intake is below recommendations in children and some staple foods, rich in inulin, may help to increase its consumption. Inulin a prebiotic dietary fibre (1) is mainly found in plant-based foods. Food reformulation has led to an increase in inulin content in various food products recently, yet there is little research on the level of inulin consumption from food sources in the diet especially in children. Aim: To explore the food sources and intake of inulin in school-aged children. 
Methods: Children were recruited between the Summer and Fall of 2019 in purposely designed child interactive science activities. Diet intake was recorded using one 24-hour recall, and with the aid of food images parents helped their child to remember the foods eaten. For the assessment of inulin a list of foods was created using food composition data reporting on inulin-type fructan content (2) (3). The amount of inulin from reported homemade and takeaway food was calculated using the mean ingredients of three recipes. Nutritics software was used to check for standard portions. 
Results: A total of 154 school-aged children, median age 7 years old, IQR (12-5) female/male ratio (61/39%) were recruited from the least deprived areas (65.3%) of Scotland. Median inulin consumption was 1.3 g/day, IQR (7.1-0). Grain and cereal products contributed over half the inulin intake (58.4%). Within this group, bread accounted for over a third of inulin (36.5%), followed by breakfast cereals and biscuits with only 13.3% and 5.1%, respectively. The second highest contributing food group was takeaway and homemade dishes which accounted for 21.7% of inulin consumption, including vegetable dishes with 9%. The vegetable, potatoes, beans and fruits food group contributed the least amount of inulin with 11.9% and 8%, respectively. 
Conclusion: In this population of school-aged children, the main food sources of inulin come from grain and cereal products and mixed meals from which carrots and onions are the likely contributing ingredients to inulin consumption. These data will help inform future intervention studies of increased dietary fibre consumption in children.

References:  
1) Roberfroid, M., Gibson, G.R., Hoyles, L., et al. (2010) Br J Nutr 104,S1-63. 
2) Biesiekierski, J.R., Rosella, O., Rose, R., et al. (2011) J Hum Nutr Diet 24(2),154-76. 
3) Whelan, K., Abrahmsohn, O., David, G.J., et al. (2011) Int J Food Sci Nutr 62(5),498-503

Food sources and inulin consumption in school-aged children

The human gut microbiota harbor trillions of microflora that can provide benefits such as protection against infection from other microbes, stimulating immune function [1] . The microbial imbalance occurs when antibiotic used and consequently this lead to a decrease in the normal flora which may result in gastroenteritis and diarrhea [2] [3]. Addressing this dysbiosis may introduce an external supplementation with beneficial micro-organisms in the form of probiotic. The term probiotic refers to “live microorganisms which when administered in adequate amounts confer a health benefit on the host” [4]. It has been proposed that probiotics have therapeutic effects for a wide range of medical conditions and helpful in maintaining a healthy and well balance gut flora. Establishing the health benefits of probiotics among health care provider (HCPs) can be positive for patients. It might contribute to a change in their dietary practices and reduce the risk of nutrition-related chronic disease. There is no data assessing knowledge and perceptions of probiotics by HCPs in KSA despite the increase in their availability to the consumers. Therefore, the aim of this study is to assess the knowledge, attitude and practices among HCPs in KSA.
A cross sectional study were conducted on Health care providers in Riyadh hospitals. The data were collected using a semi structured online questionnaire related to the definition, health benefits and the perceptions about probiotics. Data were collected and analysed using SPSS PC+ version 21.0 statistical software (SPSS Inc., Chicago, IL). 
Mean, standard deviation, frequency counts and percentages were used to present the demographic features of the participating professionals. Analysis of variance and Tukey multiple comparison test were used to evaluate probiotics knowledge considering the demography of more two (2) sub-categories. Pearson’s correlation coefficient was used to assess the possible relationship between probiotics knowledge scores and chosen demographic variables. Statistical results were considered to be significant at p≤ 0.05. 


A total of 333 health professionals participated which was about 95.7% from the original number of submitted responses. Results of the study revealed dominance of females (52.3%), aged between 30-44 years old (62.8%), mostly nurses (46.8%), and of professional experience at 10-19 years (37.5%). (85.3%)  of the participants were aware of the difinitions of probiotics, , 77.5% were having no concerns in the use of probiotics.  As per perceptions about probiotics, it was viewed as not harmful to health (64.0%); beneficial to gut health and IBS (42.6% and 32.4% respectively); and lastly viewed as not beneficial to obesity (24.9%), allergies (28.5%), and heart diseases (36.3%). With α = .05, there were significant differences on probiotics knowledge i.e., benefits to obesity (p= .006), allergy (p= .036), Crohn’s disease (p=.026) & heart disease (p= .002) when grouped according to gend

Investigation of health care providers’ knowledge of probiotics in KSA

Inter-individual variability in gut microbial metabolism and vascular response to lignans and isoflavones: the StratiPol study

Wafa Alotaibi, Paul Young Tie Yang, Wendy L. Hall, Ana Rodriguez-Mateos

Dept. of Nutritional Sciences
School of Life Courses Sciences
Faculty of Life Sciences & Medicine
King’s College London

Objectives: Evidence shows inter-individual variability in gut microbial metabolism of phytoestrogens, lignans and isoflavones, but little is known on whether this is associated with health benefits. The aim of this project is to investigate the variability in gut microbial metabolism and vascular response to lignans & isoflavones in a group of healthy individuals.

Methods: An open-label, single-arm study investigating the influence of gut microbial metabolism variability on vascular response to a (poly)phenol rich breakfast is currently ongoing in a group of healthy men and women aged 38±13 years old and with BMI 24±3 kg/m². Participants consumed a (poly)phenol rich breakfast containing 300 mg of flaxseed lignans and 22 mg of soy milk isoflavones for 3 days. Urine samples were collected for 24 hours after consumption on day 4. Flow mediated dilation, pulse wave velocity, augmentation index, blood pressure, heart rate and serum biochemistry were assessed at baseline after an overnight fast on day 1 and 24 h after the last breakfast was consumed on day 4. Urine samples were analyzed using liquid chromatography tandem mass spectrometry (LC-MS/MS) and authentic standards.

Results: Preliminary results from the first 69 volunteers (48 females vs 21 males) are presented. The main metabolites of the isoflavone daidzin (S-equol, daidzein, R,S equol 4'-sulfate, and R,S equol 7-β-D-glucuronide), and the lignan, secoisolariciresinol diglucoside ((±)-enterolactone, enterodiol, (±)-Enterolactone monosulfate and (±)-Enterolactone mono-β-D-glucuronide) gut microbial metabolites were identified and quantified in 24 h urine. In this population, 74% of the participants were non-equol producers (31% male vs 69% female) and 26% were equol producers (28% male vs 72% female). A significant decrease in systolic and diastolic blood pressure was found after 3 days isoflavone consumption only in male equol producers (120±10 to 113±8 mmHg, p=0.0046 and 77±11 to 71±12 mmHg, p=0.0024 respectively). All volunteers produced enterolactone metabolites, with a high variability in urinary levels of total and individual enterolactone metabolites. No significant differences in any vascular outcome were found between low (lower quartile ≤10 µmol, 29% men and 71% female) and high (higher quartile ≥ 44 µmol, 35% men and 65% female) enterolactone producers.

Conclusion: Sex and gut microbiota may be important factors explaining variability in the vascular response to isoflavones. More research is needed to confirm these findings, including randomised controlled trials with previous stratification by sex and polyphenol gut microbial metabotype.

Inter-individual variability in gut microbial metabolism and vascular response to lignans and isoflavones: the StratiPol study

Evidence for a role for vitamin D in metabolic-associated fatty liver disease (MAFLD) pathogenesis is conflicting. MicroRNAs (miRNAs) play an essential role in both the progression of MAFLD, and in mediating the cellular response to vitamin D including the post-transcriptional regulation of the vitamin D receptor (VDR). However, the potential roles for vitamin D regulated miRNAs in the molecular pathogenesis of MAFLD remains unexplored. With an overall objective of investigating the role of vitamin D regulated miRNAs in MAFLD, the aims of these experiments were to: measure miRNA expression in immortalised hepatocytes (HepG2 cells) in response to vitamin D alone, or in combination with lipid loading; and to examine potential functional and mechanistic effects using bioinformatics. Cells were cultured for 16hrs in charcoal-stripped serum-containing media prior to 24h treatment with either 100nM 1α,25-dihydroxyvitamin D3, or fatty acid (500μM, 1:1 oleic acid: palmitic acid), or both. miRNA samples from four biological experiments were collected using mirVANATM miRNA isolation reagents and synthesised to cDNA. TaqMan® array human miRNA A+B cards were used to measure miRNA expression in equally pooled cDNA samples. Data were examined by relative fold change, and also (in R version v4.0.3) by principal component analysis (PCA) and significance analysis of microarrays (SAM) approaches. Bioinformatic analyses included the identification of miRNA gene targets, gene-set enrichment analysis (GSEA), and functional annotation and pathway analyses; utilising the miRWALK (v3.0), DAVID (v6.8), and Reactome (v75) knowledge-bases and their associated tools. 768 miRNAs were measured across all four treatment groups. Of these, A high percentage of detected miRNAs were found increased (fold change>1.33) and decreased (fold change <0.67) relative to vehicle-treated cells (56.0%, 25.3% and 56.5% in vitamin D, fatty acid, and co-treated cells respectively). However, SAM analysis across groups only identified 70 miRNAs as significantly up-regulated, and 5 down-regulated at delta=0.822 and a median false discovery rate of 0.112. Utilising miRWALK and GSEA, 23 pathways found significantly altered under all 3 different treatment conditions, which were interesting signalling pathways such as the PI3K-Akt, FoxO, AMPK, MAPK and p53 (P<0.001) regulated. In summary, these experiments identify miRNAs regulated by vitamin D and lipid loading in immortalised hepatocytes. Bioinformatic results highlight signalling pathways with critical roles in insulin signalling known to be dysregulated in diabetes and MAFLD. Ongoing work is integrating these data with analyses from analogous experiments in immortalised hepatic stellate cells (LX-2). From this, a subset of candidate miRNAs will be followed up for independent verification of functional and mechanistic effects.

Characterisation of microRNAs regulated by vitamin D and lipid loading in immortalised hepatocytes

The gastrointestinal tract is amongst the most densely populated ecosystems on earth, home to an abundant and diverse collection of microbial species, not just bacteria but also Archaea, viruses, fungi etc. Significant interest in this area has been sparked in recent years by an appreciation of the importance of the gut microbiota in the health of the host, and this has led to the microbiome of humans and animals being investigated in depth. Studies have drawn links between host nutrition, the gut microbiota and metabolic and immune functions, which in consequence affect physiology.



The first investigations of the structure of the microbiota were culture-based, however the introduction of sequencing technologies such as Sanger sequencing and subsequent development of 454 sequencing and metagenomics approaches have become the mainstay of microbiota studies. The creation of reference databases and analysis software has provided invaluable tools for assigning taxonomy to generated data. Great progress in characterising the structure of the microbiome recently has paved the way for ongoing and future studies on the functional interactions between the microbiota and the host. Studies on the function of the microbiota are critical in understanding the role of the microbiota in human homeostasis and disease pathogenesis.



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