Excess vitamin intake during pregnancy leads to obesogenic phenotypes, and folic acid accounts for many of these effects in male, but not in female, offspring. These outcomes may be modulated by another methyl nutrient choline and attributed to the gut microbiota. Pregnant Wistar rats were fed an AIN-93G diet with recommended vitamin (RV), high 10-fold multivitamin (HV), high 10-fold folic acid with recommended choline (HFol) or high 10-fold folic acid without choline (HFol-C) content. Male and female offspring were weaned to a high-fat RV diet for 12 weeks post-weaning. Removing choline from the HFol gestational diet resulted in obesogenic phenotypes that resembled more closely to HV in male and female offspring with higher body weight, food intake, glucose response to a glucose load and body fat percentage with altered activity, concentrations of short-chain fatty acids and gut microbiota composition. Gestational diet and sex of the offspring predicted the gut microbiota differences. Differentially abundant microbes may be important contributors to obesogenic outcomes across diet and sex. In conclusion, a gestational diet high in vitamins or imbalanced folic acid and choline content contributes to the gut microbiota alterations consistent with the obesogenic phenotypes of in male and female offspring.
Objectives Excess gestational folic acid and insufficient choline intakes as observed in the North American populations may increase the risk of obesity in offspring. It is well-established that adverse health outcomes may arise due to shifts in the gut microbial communities, but whether high vitamin intakes or an imbalance between methyl nutrients contributes to gut microbiota alterations is unclear. The objective of this research was to determine the gut microbiota composition of male and female offspring in relation to the vitamin composition of the gestational diet. Methods Pregnant Wistar rats (n = 10/group) were fed the AIN-93G diet with either the recommended vitamin (RV), high multivitamin (HV), high folic acid (HFol) or high folic acid without choline (HFol-C) content. Male and female offspring were weaned to a high-fat control diet for 12 weeks. Fecal samples were collected from the colon upon termination for gut microbiota profiling by 16S rRNA sequencing and data analyses in QIIME2. Results The overall gut microbial communities as assessed by unweighted UniFrac distances differed among the gestational diet groups for male (PERMANOVA P = 0.04) and female (PERMANOVA P = 0.05) offspring. The covariates gestational diet and sex predicted the gut microbiota differences in the offspring (Q2 = 0.07 in Songbird) whereas diet alone resulted in overfitting of the multinomial regression model (Q2 < 0). High ranked features from the natural log-ratios of microbial abundance were Shigella, Clostridiales, Clostridiaceae for HV, and Odoribacter, Akkermansia muciniphila, Blautia for both HFol and HFol-C compared to RV. Low ranked features were Odoribacter for HV, Clostridiaceae and Clostridiales for HFol, and Bifidobacterium, Allobaculum, Lactobacillus vaginalis for HFol-C compared to RV. In male offspring, Lactobacillus vaginalis, Sutterella and Clostridiales were high ranked and Odoribacter was low ranked compared to female offspring. These differentially abundant microbes may be important contributors to obesity across diet and sex. Conclusions Increased vitamin content or an imbalance between folic acid and choline in the gestational diet leads to a shift in the gut microbiota composition in the offspring toward obesity. These effects differed by sex. Funding Sources Utah Agricultural Experiment Station and USU Research Catalyst. UNM supported by USU URCO.
Objectives The objective of this study was to investigate differences in live and dead bacteria counts a month before the expiration date and right after the expiration date and observe any differences in bacteria counts between types of yogurt, brands, and stores tested. Methods Two yogurt containers with the same expiration date were collected for each brand at three stores. The yogurts collected were three national brands of Greek yogurts and five regular yogurts. The first container was tested one month prior to expiry. The second container was tested at the expiration date. After being thoroughly mixed, triplicate 50–75 mg of yogurt were sampled from each carton. Yogurt samples were diluted 9-fold with freshly prepared peptone water. A second dilution was prepared by mixing 25 µL of the yogurt peptone mix with 975 µL of freshly prepared sterile PBS. The samples were incubated for 15 min. at 37°C with 1 µL SYBR green and 5 µL propidium iodine and iced for 1 hour in the dark. 200 µL of dyed yogurt were diluted with 800 µL of PBS and analyzed by flow cytometer (BD Accuri™ C6 Cytometer). Each yogurt sample was run in triplicate on slow speed setting. Flow cytometer parameters were set to differentiate live and dead bacteria based on fluorescence as well as yogurt matrix artifacts. The data was analyzed by a least-squares fit model to test for significance among variables. Results Greek yogurts had significantly higher live and dead bacteria counts compared to regular yogurts. The Greek yogurts differed amongst themselves. Brand 3 had significantly lower live counts than brand 2 and 1. Brand 1 had significantly higher counts of dead bacteria compared to the other two. While differences between Greek yogurt are statistically significant, they are most likely not clinically significant. For the regular yogurt live and dead counts there were no significant differences. No significant differences were observed based on the timing of yogurt sampling or the store the yogurt came from. Conclusions Preparing Greek yogurt involves more straining and whey removal and yields a higher yogurt concentration, so it is no surprise that Greek yogurts have higher bacteria counts. This study demonstrates that when considering yogurt for its probiotic qualities, the most relevant factor is the type of yogurt. Yogurt brand, store, or when it is sampled have little effect on the probiotics count. Funding Sources USU.
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