Introduction/Background Research indicates nutrition and environment in the first year of a child's life are crucial in their development and growth and can contribute to lower chances of developing obesity and other health concerns. Key factors that can determine these outcomes include the bacteria and resulting short chain fatty acids (SCFAs) present in the gut. This composition may be affected by feeding modality (formula feeding vs breastfeeding), exposure to the mother’s microbiota, weight status of the child, and type of delivery. This research aims to identify the impact of infant feeding modality on toddlers' fecal fermentation profile, and if there are associations between weight status and microbiome, fecal fermentation profile. Methods/ Procedures Participants (n=40) were recruited during well-child pediatric appointments at ETSU’s Pediatric primary care clinic. Researchers explained the requirements of the study and participants were provided with a 90-question food frequency questionnaire (FFQ) for children ages 2-7, including 90 questions and asks about a child's typical intake over the previous 6-month period. The food list was developed from NHANES III dietary recall data. The child’s history was obtained, including current age, birth length and weight, delivery type (C-section or vaginal), feeding method (breast, bottle fed, or both) and duration. The child’s weight and height were obtained, and body mass index (BMI) calculated. Participant-provided stool samples were freeze-dried and ground, and SCFAs were extracted using a procedure developed by Schwiertz et al. that was modified. One mL of the SCFA extraction solution, containing Oxalic acid (0.1 mol/L), Sodium Azide (40 mmol/L), and Caproic acid (0.1 mmol/L) (internal standard) was added to 80 mg of a freeze-dried stool sample in a 16 x 100 mm disposable culture tube, and analyzed using a Shimadzu GC2010 gas chromatograph with SigmaAldrich ZB-Wax Plus capillary column. Samples were run in duplicate, and values for each participant were averaged. Data analysis was generated using SAS software, Version 9.4 of the SAS System, Copyright © 2013 SAS Institute Inc. Results Initial findings showed no significant differences in the SCFA composition of obese vs non-obese toddlers in the sample. However, there were significant differences in the amount of specific SCFAs (isobutyrate, isovaleric acid, and octanoic acid) in toddlers who were formula fed as infants versus toddlers who were breastfed, and those fed a combination of breastmilk, and formula (p < 0.05). Further analysis will determine if these initial results may be contributed to overall dietary intake, and more specifically fiber intake.

The Fecal Fermentation Profile of Twins and Infants with Opioid Exposure Dawson Brown-Ezell Michelle Johnson PhD, RD, LDN W. Andrew Clark PhD, RD Amy Wahlquist, PhD Introduction: The gut microbiome is believed to have a significant impact on health throughout the lifespan, and the influence of infant nutrition and other environmental factors are of particular interest in its development. The aim of this research project was to learn more about the microbiome and short chain fatty acid (SCFA) composition of toddlers of differing weights, considering birth history, environment, and diet. In East Tennessee, opioid misuse is a growing issue, and a number of participants in this study were exposed in utero. We also hoped to identify related effects on infant’s SCFA composition. Finally, it has been concluded that twins share a variety of traits, but much about their microbiome is unknown. With several pairs of twins in the sample, we aimed to identify any associations with SCFAs in this group. Methods: With informed consent, the child’s history was obtained, including age, birth length and weight, delivery type (C-section or vaginal), and feeding method (breast, bottle fed, or both). The child’s current weight, height, and BMI %ile were determined. Caregivers completed the 90-question Block Questionnaire for Ages 2-7 Kids food frequency questionnaire, and results were analyzed by Berkeley Analytics Inc (dba NutritionQuest). Participant-provided stool samples were freeze-dried and ground, and SCFAs were extracted and analyzed by content and concentration. Data analysis was generated using SAS software, Version 9.4 of the SAS System, Copyright © 2013 SAS Institute Inc. Results: Nine SCFAs were measured in duplicate, and the concentrations averaged. Statistical analysis included comparisons of SCFAs related to factors including weight status, infant feeding modality, twin status, and intrauterine drug exposure, and significance determined with a p value < 0.05. Results did not identify significant differences in individual SCFA concentrations between obese and non-obese toddlers, however concentrations of isobutyrate, isovaleric acid, and octanoic acid were greater in toddlers who were formula fed as infants versus toddlers who were breastfed, and those fed a combination of breastmilk, and formula. Analysis further revealed a higher mean concentration of caproic and propionic acid in twin subjects. Of particular interest, toddlers with a history of opioid exposure had higher mean concentrations of isovaleric and octanoic acids, but less isocaproic acid when compared to those who were not drug exposed. Further analysis will help determine if these findings may be related to nutrient intake, in particular dietary fiber intake.

Introduction: The gut microbiome is believed to have a significant impact on health throughout the lifespan, and the influence of infant nutrition and other environmental factors are of particular interest in its development. The aim of this research project was to learn more about the microbiome and short chain fatty acid (SCFA) composition of toddlers of differing weights, considering birth history, environment, and diet. In East Tennessee, opioid misuse is a growing issue, and a number of participants in this study were exposed in utero. We also hoped to identify related effects on infant’s SCFA composition. Finally, it is known that twins share a variety of traits, but much about their microbiome is unknown. With several pairs of twins in the sample, we aimed to identify any associations with SCFAs in this group. Methods: With informed consent, the child’s history was obtained, including age, birth length and weight, delivery type (C-section or vaginal), and feeding method (breast, bottle fed, or both). The child’s current weight, height, and BMI percentile were determined. Caregivers completed a 90-question Block Questionnaire for Ages 2-7 Kids food frequency questionnaire, and results were analyzed by Berkeley Analytics Inc (dba NutritionQuest). Participant-provided stool samples were freeze-dried and ground, and SCFAs were extracted and analyzed by content area % and concentration ppm. Data analysis was generated using SAS software, Version 9.4 of the SAS System, Copyright © 2013 SAS Institute Inc. Results: Nine SCFAs were measured in duplicate, and the concentrations averaged. Statistical analysis included comparisons of SCFAs related to factors including weight status, infant feeding modality, twin status, and intrauterine drug exposure, and significance determined with a p value < 0.05. Results did not identify significant differences in individual SCFA concentrations between obese and non-obese toddlers, however concentrations of isobutyrate, isovaleric acid, and octanoic acid were greater in toddlers who were formula fed as infants versus toddlers who were breastfed, and those fed a combination of breastmilk, and formula. Analysis further revealed a higher mean concentration of caproic and propionic acid in twin subjects. Of particular interest, toddlers with a history of opioid exposure had higher mean concentrations of isovaleric and octanoic acids, but less isocaproic acid when compared to those who were not drug exposed. Further analysis will help determine if these findings may be related to nutrient intake, in particular dietary fiber intake.

The cell wall provides a complex and rigid structure to the plant for support, protection from environmental factors, and transport. It is mainly composed of polysaccharides, proteins, and lignin. Arabinoxylan (AX), pectin (P), and cellulose (C) are the main components of cereal cell walls and are particularly concentrated in the bran portion of the grain. Cereal arabinoxylans create networks in plant cell walls in which other cell wall polysaccharides are imbedded forming complex matrices. These networks give an insolubility profile to plant cell wall. A previous study in our lab showed that soluble crosslinked arabinoxylan with relatively high residual ferulic acid from corn bran provided advantageous <i>in vitro </i>human fecal fermentation products and promoted butyrogenic gut bacteria. In the present work, arabinoxylan was isolated from corn bran with a mild sodium hydroxide concentration to keep most of its ferulic acid content. Highly ferulated corn bran arabinoxylan was crosslinked to create an insoluble network to mimic the cereal grain cell wall matrices. Firstly, arabinoxylan film (Cax-F), pectin film (P-F), the film produced by embedding pectin into arabinoxylan networks (CaxP-F), and cellulose embedding arabinoxylan matrices (CaxC-F), and embedding the mixture of cellulose and pectin into arabinoxylan networks (CaxCP-F) were fabricated into simulated plant cell wall materials. Water solubility of films in terms of monosaccharide content was examined and revealed that Cax-F was insoluble, and P-F was partially insoluble, and nanosized pectin and cellulose were partially entrapped inside the crosslinked-arabinoxylan matrices. In a further study, these films were used in an <i>in vitro </i>human fecal fermentation assay to understand how gut microbiota access and utilize the different simulated plant cell walls to highlight the role of each plant cell wall component during colonic fermentation. <i>In vitro </i>fecal samples, obtained from three healthy donors were used to ferment the films (Cax-F, P-F, CaxP-F, CaxC-F, and CaxCP-F) and controls (free form of cell wall components -Cax, P and C). The fabricated films that were compositionally similar to cell walls were fermented more slowly than the free polysaccharides (Cax and P). Besides, CaxP-F produced the highest short chain fatty acids (SCFA) amount among the films after 24 hour <i>in vitro </i>fecal fermentation. Regarding specific SCFA, butyrate molar ratio of all films was significantly higher than the free, soluble Cax and P. 16S rRNA gene sequencing explained the differences of the butyrate proportion derived from specific butyrogenic bacteria. Particularly, some bacteria, especially in a butyrogenic genera from Clostridium cluster XIVa, were increased in arabinoxylan films forms compared to the native free arabinoxylan polysaccharide. However, no changes were observed between P and P-F in terms of both end products (SCFA) and microbiota compositions. Moreover, CaxP-F promoted the butyrogenic bacteria in fecal samples compared with pectin alone, arabinoxylan alone, and the arabinoxylan film. Differences in matrix insolubility of the film, which was high for the covalently linked arabinoxylan films, but low for the non-covalent ionic-linked pectin film, appears to play an important role in targeting Clostridial bacterial groups. Overall, the cell wall-like films were useful to understand which bacteria degrade them related to their physical form and location of the fiber polymers. This study showed how fabricated model plant cell wall films influence specificity and competitiveness of some gut bacteria and suggest that fabricated materials using natural fibers might be used for targeted support of certain gut bacteria and bacterial groups.