Progress 03/15/17 to 03/14/22
Outputs
Target Audience:This project is targeting the scientific community and product developers in the food/nutrition industry who are seekingsolutions to enhance intestinal health by incorporating and delivering butyrate into food matrices. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?An undergraduate student was trained for the anlaysis of samples collected from the piglet study. How have the results been disseminated to communities of interest?Twoposter presentations have been accepted for the Nutrition 2022 conference. There have been several meetings with companies in both human and animal foods. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The overall goal of this study is to encapsulate tributyrin (TB) using various techniques to control its release in the gastrointestinal (GI) tract and to reduce its bitterness perception for oral delivery to benefit populations with various GI issues. Aims 1, 2, and 3: Several encapsulation techniques have been employed to form the encapsulated TB including spray-drying with two and three fluid nozzles (2FN and 3FN, respectively), inclusion complexation and cross-linking of biopolymers. Various wall materials such as whey protein isolate (WPI), gamma-cyclodextrin (CD) and sodium alginate (AL) were selected based on their functionalities. The retention of TB and surface oil content wasquantified to compare encapsulation efficiency. Physical properties of the capsules, which affect the behavior of the capsules, were characterized. The properties included particle size, morphology, crystallinity, and molecular interaction. Most importantly, in-vitro release of TB in a simulated GI condition and the ability to mask bitterness were evaluated. It was found that the surface oil content of spray dried TB/WPI microcapsules can be significantly reduced using 3FN or by incorporating anhydrous milk fat, thus the encapsulation efficiency and reconstituted emulsion stability increased. TB and CD formed a complex and the complex was successfully spray-dried with full retention of TB at 1:2 molar ratio of TB: CD. The TB/CD complex was highly fermentable to produce butyrate using ileum and colon contents collected from piglets. TB/CD/AL beads also provided full retention of TB and showed the most promising results in in-vitro digestion: It exhibited a controlled slow release of TB with 30 % TB retention after twohours in stomach and twohours in small intestinal conditions as well as successfully masked the bitterness of TB in oatmeal, making it a potential functional ingredient for the oral delivery of TB/butyrate to colon. Overall, different encapsulation methods provide TB contained capsules with different properties that can be used for various applications. For the future, TB/CD/AL microcapsule should be developed for enhanced encapsulation, delivery of butyrate as well as for improved masking bitterness and grainy mouthfeel when incorporated in food matrices. Aim 4:Two-day old piglets (N=125) were randomly allocated to three dietary treatments: formula alone (CON), formula with 9.0 mM gamma-cyclodextrin (CD)or 9.0 mM gamma-cyclodextrin/ tributyrin complex (CDTB). Body weight (BW) and disease activity index was assessed daily. On study day twelve, each group was further divided into colitis (CON+, CD+, CDTB+) and non-colitis (CON-, CD-, CDTB-). Colitis piglets were dosed with 1.25 g/kg body weight DSS (20% solution in water) each morning for five consecutive days (days 12 to 16 of study). BW and formula intake were measured daily. Piglets were euthanized on study days 17 and 25. Blood was obtained via cardiac-puncture and later centrifuged for plasma separation. Small (SI) and large intestines (LI) were excised, length and weight measurements obtained, and sections of intestine and ileum, ascending (AC), and descending colon (DC) contents were snap-frozen in liquid nitrogen and were stored in an ultra-low freezer prior to analyses. DNA, 165 rRNA, VFA (volitile fatty acids) were analyzed. Supplementing γ-cyclodextrin/tributyrin to formula resulted in:1) Slower weight gain relative to CON animals, especially in CDTB+ group, 2) No amelioration of the DSS-induced structural changes in the DC, 3) Changes in ileum and AC morphology, 4) Increased secretion of the protein TNF-α in the descending colon on D17 which was accompanied by higher gene expression on day 25 when compared to controls. Also, the encapsulated CD/TB modulates the bacterial community throughout the piglet intestinal tract. And colitis influences both the microbiota composition and VFA concentrations. Overall, the encapsulation of TB in CD was achieved yet the controlled released of butyrate needs to be improved especially for tansverse and descending cololon areas to improve the efficacy of the encapsulation tributyrin. Futher studies on encapsulation such as using cross linked sodium alginate as an additional wall material is needed.
Publications
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
Marcia H. Monaco, Youngsoo Lee and Sharon M. Donovan. 2022. Encapsulated Tributyrin and Gamma-Cyclodextrin Does Not Ameliorate Intestinal Immune Response During DSS-Induced Colitis in Neonatal Piglet. Nutrition 2022, American Society of Nutrition.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
Mei Wang, Marcia H. Monaco, Youngsoo Lee and Sharon M. Donovan. 2022. Encapsulated Tributyrin With Gamma-Cyclodextrin Modulates the Gut Microbiota
Composition in a DSS-Challenge Piglet Model. Nutrition 2022, American Society of Nutrition.
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Progress 03/15/20 to 03/14/21
Outputs
Target Audience:This project is targeting the scientific community and product developers in the food/nutrition industry who are seekingsolutions to enhance intestinal health by incorporating and delivering butyrate into food matrices. Changes/Problems:A no-cost extension was requested due to the delays in pig sample analysis caused by COVID19. What opportunities for training and professional development has the project provided?A graduate student (Ph.D.) and several undergraduate students have been trained during this period. The graduatestudent has been trained to create TB-CD complex and operate a spray dryer. The undergradute students participated in the pig feeding study and analysis of the samples collected from the pig feeding study. The graduate student analyzed the data and presented the results at a scientific meeting. How have the results been disseminated to communities of interest?The results from a part of the project have been presented at a scientific meetings (Institute of Food Technologistsmeeting in 2020. Two peer reviewed manucripts are being prepared. What do you plan to do during the next reporting period to accomplish the goals?The data from the piglet study will be further analyzed in the next reporting period. The feeding is completed but the samples need to be analyzed. Manuscripts will be prepared and submitted. The final report will be prepared and submitted. On-Going Analyses: Analyses Estimation for completion Ileum and AC morphology End of the summer, 2021 AC and DC RNA isolation for cytokine gene expression by qPCR Mid-summer, 2021 Quantification of ileal and AC total bacteria, Roseburia spp. & Eubacterium rectale, Faecalibacterium prausnitzii and Enterobacteriaceae, and bacterial butyryl-CoA:acetate CoA-transferase gene expression by qPCR End of the summer, 2021 Microbiota taxonomic analysis End of the summer, 2021 Manuscripts Fall 2021
Impacts
What was accomplished under these goals?
For Aim 4: Animal Protocol All animal care and experimental procedures were in accordance with the NRC Guide for the Care and Use of Laboratory Animals [NRC, 1996] and were approved by the University of Illinois Institutional Animal Care and Use Committee (protocol #20188). Two-day-old piglets (n=128) were obtained from the University of Illinois Swine Research Center and were transported to the animal facility and housed in a specialized neonatal rearing system. Piglets were randomized into three experimental diets: 1) Formula alone (control, CON); 2) Formula + 9 nM encapsulated g-CD/TB (CDTB); or 3) Formula + 9 nM g-CD (CD). The study was conducted in fourreplicates of 32 piglets with all dietary treatments represented in each replicate. The study was ran from February through August 2019. Three animals from Replicate 2 were removed from the study due to failure to thrive. Piglets were fed a commercially available sow-milk replacer diet (Advance Baby Pig Liqui-Wean, Milk Specialties, Dundee, Illinois), that was reconstituted daily as 20% solids. Both g-CD/TB and CD were added to the formula at the time of its preparation. Piglets were weighed each morning to assess growth and to determine formula intake. Formula was provided at 300 and 325 mL per kg body weight (BW) from study day (D) 0-5 and 6-25 of the study, respectively. Milk was automatically dispensed using a pump as elevenequally-spaced meals per day. On days 12-16, half of the piglets in each dietary group were dosed once daily with DSS (MW, 36,000-50,000; Thermo Scientific, Tewksbury, MA) by providing 1.25 g/kg BW. The DSS was prepared as a 20% solution in water and poured into feeding bowl at the concentration of 6.25 mL/ kg BW prior to the first morning feed. On study days 17 and 25, piglets were euthanized for sample collection to examine dietary treatment effect after the cessation of DSS treatment and in the recovery phase. * Body weight was measured and blood samples were collected. * Intestinal Analysis - DC Histopathological Evaluation: Formalin-fixed sections were processed into paraffin blocks, sectioned, placed on glass slides and stained with H&E at the University of Illinois, College of Veterinary Medicine. Digital images (20x) of sections will be captured using the NanoZoomer Slider Scanner (Hamamatsu Corporation, Bridgewater, NJ, University of Illinois Institute for Genomic Biology Microscopy and Imaging Facility). The quantitative histological inflammation grading used to access the degree of colitis was adapted from Dieleman et al. [1998]. Four independent parameters were graded as described in Table 1 below, and the score of each parameter was multiplied by a factor reflecting the percentage of tissue involvement to yield scoring ranges of 0-27.Two scorers, blinded to experimental treatments, were training with the use of images representing for each scoring grade. One intestinal cross-section from each animal was divided in three sections, and each section was evaluated separately. The average of the three sections represented the scoring grade for each animal. Table 1. Feature graded Grade Epithelium Infiltration Crypt Damage % Extent Affected 0 - Epithelial lining is fully intact. - Uniformity is present - Cuffs are densely packed which maximizes surface area Normal: - No or minimal cell Infiltration. - Uniformly packed cuffs with goblet cells present Normal: - No Crypt Damage - Crypts (cuffs) are uniform and densely packed with goblet cells 0%: - The circumference of the section is uniform throughout - Intact epithelium and cuffs over the diameter of the section 1 - Mild to Moderate loss of the uniformity of the epithelial lining. - Mild loss of epithelial continuity and fractured regions Mild Infiltration: - Infiltration of immune cells within the mucosa and lamina propria. - Slight elongation of crypts (hyperplasia) with minimal goblet cell loss Basal crypt mildly damaged: - Some disarray in uniformity can be seen. - Crypts are filled with mucus 25%: - Roughly 1/4th of the circumference of the section is affected with the disease. - The remaining ¾ of the section is uniform with minor structural abnormalities present 2 - Moderate to Severe Loss of the Epithelial Lining - Not Uniformed, many fractures and breakages in the lining exist - Not continuous Moderate to marked infiltration : - Infiltration in the mucosa, lamina propria and submucosa - Crypt elongation (hyperplasia) is significant. - Significant goblet cell loss Basal crypt moderately or severely damaged: - Crypt structure has morphologically changed. - Non-parallel crypts, variable crypt diameters, bifurcation and branched crypts 75%: - Roughly 3/4 of the circumference of the section is affected with the disease (75% is in disarray). - A small portion of the section is uniform 3 - Architectural Destruction of the Epithelium throughout. - Transmural reach. - Transmural destruction of intestinal cell layers - No visible crypts or goblet cells. Transmural damage: - Entire crypt and epithelium lost 100%: - Extent of disease is visible in the section's entirety. - Transmural damage is observed Intestinal Cytokine Concentration: IL-6 and TNF-α concentration in AC and DC mucosal were analyzed using the same ELISA kit described for plasma. Intestinal homogenates were prepared by homogenizing 0.5 g of tissue in 1 mL Hank's balanced solution with protease inhibitor cocktail as previously described [Lackeyram et al., 2017]. Ileal, AC, and DC Dry Matter and Volatile Fatty Acid (VFA) Concentrations: Dry matter was measured by difference in weight before and after lyophilization. The VFA (acetate, butyrate, propionate, isobutyrate, valerate and isovalerate) concentrations were analyzed by gas chromatography as described previously [Li et al., 2012]. Microbiota Analysis: DNA was isolated from ileum (N=58), ASC (N=57) and DSC (N=55) content from D25 animals by combination of the QIAamp Fast DNA Stool Mini Kit (Qiagen, Valencia, CA, USA) with bead beating on the FastPrep-24 System (MP Biomedicals, Carlsbad, CA, USA), as previously described [Li et al., 2012]. The concentration of DNA was measured with a NanoDrop 1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). PCR amplification and sequencing of the V3-V4 region of 16S rRNA genes were performed at the DNA Services Lab, University of Illinois as described by Monaco et al. [2018]. Bioinformatics and statistics were conducted following protocol described previously, except that demultiplexed reads were processed using QIIME 2 [Boyle et al., 2019]. Summary of Results: CDTB and colitis led to lower body weight gain compared to CON and CD-fed animals. Small intestinal weight and length were affected by colitis and diet, respectively on D17, while small and large intestine length were affected by diet and colitis, respectively, on D25. Supplementation with CD or CDTB did not impact the DC morphological changes due to colitis. Tissue disruption was mild on D17 and severe on D25. Colonic IL-6 and TNF-α concentrations were mostly affected by DSS-induced colitis on D17 in the ASC and DSC. IL-6 concentration in the ascending colon was significantly lower in CD and CDTB non-colitis animals. CDTB supplementation increased acetate and butyrate, isobutyrate, isovalerate, and valerate concentration relative to CON in the ileum, AC and DC, respectively. VFA concentration increased from day 17 to day 25 in both AC and DC. CDTB supplementation influenced alpha diversity of D25 ileum and AC, while presence of colitis changed the microbiota biodiversity in the DC. Diet supplementation with CD or CDTB the overall composition of the ileal microbiota when compared to CON, while microbiota in the AC and DC were influenced by colitis. (CDTB: cyclodextrin-tributyrin complex, CON: control, AC: Ascending colon, DC: Descendin colon, and VFA: Volatile fatty acid)
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2020
Citation:
Xueqian Shi and Youngsoo Lee. 2020. Effect of anhydrous milk fat (AMF) on the encapsulation efficiency of tributyrin using spray drying. IFT Annual meeting 2020.
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Progress 03/15/19 to 03/14/20
Outputs
Target Audience:This project is targeting the scientific community and product developers in the food/nutrition industry who are seeking solutions to enhance intestinal health by incorporating and delivering butyrate into food matrices. Changes/Problems:A no cost extension was requested due to the delays in pig sample analysis. What opportunities for training and professional development has the project provided?A graduate student (Ph.D.) and several undergraduate students have been trained during this period. The graduate studenthas been trained to create TB-CD complex and operate a spray dryer, gas-chromatography (GC), X-ray crystallography(XRD) and scanning electron microscope (SEM). The grduate student and undergradute students also participated in piglet feeding and analysis. The graduate student analyzed the data and presented the work at a scientific meeting. Also, the undergraduatestudents were trained to prepare samples from the spray drying and they participated in the piglet feeding study. How have the results been disseminated to communities of interest?The results from this part of the project have been presented at two scientific meetings (Institute of Food Technologists meeting in New Orleans, LA and the FASEB Summer Conference in West Palm, FL). One peer reviewed manucript was published and the second one is being prepared. What do you plan to do during the next reporting period to accomplish the goals?The data from the piglet study will be analyzed in the next reporting period. The feeding is completed but the samples need to be analyzed. The data will be collected and analyed. Manuscripts will be prepared and submitted. The final report will be prepared and submitted.
Impacts
What was accomplished under these goals?
Introduction: The objective of Aim 4 of the grant was to determine the efficacy of encapsulated γ-CD/TB complex on gastrointestinal health and inflammation using the dextran sodium sulfate (DSS)-challenge piglet model. The animal study was conducted in fourreplicated between February and August 2019. Materials & Methods: Two-day old piglets (N=125) were randomly allocated to threedietary treatments: formula alone (CON), formula with 9.0 mM gamma-cyclodextrin (CD) or 9.0 mM gamma-cyclodextrin/ tributyrin complex (CDTB). Body weight (BW) and disease activity index was assessed daily. On study day twelve, each group was further divided into colitis (CON+, CD+, CDTB+) and non-colitis (CON-, CD-, CDTB-). Colitis piglets were dosed with 1.25 g/kg body weight DSS (20% solution in water) each morning for five consecutive days (days 12 to 16 of study). BW and formula intake were measured daily. Piglets were euthanized on study days 17 and 25. Blood was obtained via cardiac-puncture and later centrifuged for plasma separation. Small (SI) and large intestines (LI) were excised, length and weight measurements obtained, and sections of intestine and ileum, ascending (AC), and descending colon (DC) contents were snap-frozen in liquid nitrogen and were stored in an ultra-low freezer prior to analyses. Sample Analysis:Plasma, and AC and DC IL-6 and TNF-α concentration were analyzed using commercially available ELISA kits (R&D Systems, Minneapolis, MN). Intestinal homogenates were prepared by homogenizing 0.5 g of tissue in 1 mL Hank's balanced solution with protease inhibitor cocktail. Ileum, ascending colon, and rectal contents were analyzed for volatile fatty acids (VFA), and DNA was extracted from those samples for bacterial 16S RNA gene sequencing. Statistical Analysis:Analysis of variance (ANOVA) was conducted using the MIXED procedure in SAS 9.4 (SAS Inst. Inc., Cary, NC) to differentiate dietary and colitis treatment effects on young pigs. Depending on the outcome, one of two statistical models were used: 1) Data collected at a single time-point (i.e., cytokine concentration, organ weights) were analyzed by a simple 1-way ANOVA, and 2) Data collected from the same animal on more than one occasion (i.e., BW) were analyzed as a 2-way, repeated-measures ANOVA. Data were reported as means ± SEM, and statistical significance was set at p < 0.05. Results: There was no main effect of dietary treatment and colitis on BW. However, the interaction between day and colitis indicated that BW of animals with colitis was significantly lower than non-colitis from days 23 to 25. Total BW gain throughout the study was lower in the CDTB compared to CON and CD groups. Small and large intestine weight and length were affected by day and colitis. Day 17 measures differed significantly from all day 25 variables. The presence of colitis had an overall negative effect on the SI weight. A diet and colitis interaction was found in that LI length in the CD, and CDTB groups differed significantly from CON. Local secretion of pro-inflammatory cytokines IL-6 and TNF-α were measured in colonic homogenates. In the AC, IL-6 concentration was significantly higher in day 17 vs. day 25 samples suggesting an acute inflammation. In DC, the interaction of diet and colitis was significant and indicated that IL-6 concentration in the CD and CDTB colitis groups were higher than the non-colitis. The same effect was not observed in the control animals. D17 AC from colitis pigs had higher TNF-α concentration than day 17 non-colitis, and day 25 colitis and non-colitis piglets, while in the DC TNF-α levels were significantly higher in colitis vs. non-colitis samples on both days 17 and 25. No diet effect was observed on cytokine levels. VFA concentration was affected by both diet and the presence of colitis. Butyrate concentration in the ileum of CDTB animals was higher than CD and CON. In the AC, butyrate and valerate concentration was higher in the colitis vs. non-colitis. A similar result for valerate was found in DC. Ongoing analysis: The sequencing of the 16S RNA genes for ileum, ascending, and descending colon samples has been completed and currently being curated and analyzed. Besides, we are assessing intestinal morphology and scoring the presence of the inflammation due to colitis. Other analyses that will start in the fall are cytokine gene expression in the intestine by PCR.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Shi, X. and Lee, Y. 2019. Encapsulation of tributyrin with whey protein isolate prepared by 2- and 3-fluid nozzle spray drying. IFT Annual Meeting, New Orleans, LA June 2019.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Monaco, M.H., Shi, X., Lee, S.Y., Wang, M., Perkowski, C., Lee, Y. and Donovan, S.M. 2019. Screening the acceptability and the biological effects of encapsulated tributyrin and gamma-cyclodextrin complex on piglet intestinal development. FASEB Summer Conference The Origins and Benefits of Biologically Active Components, West Palm, FL July 2019.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2020
Citation:
Shi, X. and Lee, Y. 2020. Encapsulation of tributyrin with whey protein isolate (WPI) by spray-drying with a three-fluid nozzle. Journal of Food Engineering. 281:109992.
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Progress 03/15/18 to 03/14/19
Outputs
Target Audience:This project is targeting the scientific community and product developers in the food/nutrition industry who are seeking solutions to enhance intestinal health by incorporating and delivering butyrate into food matrices. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A graduate student (Ph.D.) and several undergraduate students have been trained during this period. The graduate student has been trained to create TB-CD complex and operate a spray dryer, gas-chromatography (GC), X-ray crystallography (XRD) and scanning electron microscope (SEM). The grduate student also participated in piglet feeding and analysis process. The graduate student analyzed the data and presented the work at a scientific meeting. Also, the undergraduate students were trained to prepare samples from the spray drying and they participated in thepiglet feeding study. How have the results been disseminated to communities of interest?The results from this part of the project have been presented at a scientific meeting (the Institute of Food Technologists meeting inNew Orleans). Two peer reviewed publications are being prepared. What do you plan to do during the next reporting period to accomplish the goals?An additional piglet feeding study is being conducted with more piglets. Also, a sensory test will be conducted with encapsulted CD/TB in a food matrix.
Impacts
What was accomplished under these goals?
During this project period, encapsulated g-CD/TB complex was tested with piglets. Introduction: Human milk and most infant formulas contain prebiotics that are fermented by gut bacteria to volatile fatty acids (VFA). One VFA, butyrate, is a primary energy source for colonocytes and modulates gut inflammation. As such, butyrate has been proposed as an intervention to improve intestinal health. However, its sensory characteristics limit its use as an oral supplement. Administering tributyrin (TB) rather than butyrate and encapsulating it in gamma-cyclodextrin (g-CD) presents an opportunity to deliver efficacious doses of butyrate by mouth. Herein, we tested the tolerance and bioactivity of oral g-CD/TB supplementation on markers of intestinal development and volatile fatty acid (VFA) concentrations in piglets. Methods: Two-day-old piglets (n=40) were randomized to receive one of five diets for 21 days: formula alone (CON); formula + 4.5 mM g-CD (CD-LOW); formula + 4.5 mM g-CD/TB (CD/TB-LOW); formula + 9.0 mM g-CD (CD-HIGH); or formula + 9.0 mM g-CD/TB (CD/TB-HIGH). The doses were selected to approximate gut VFA concentrations, based on previous in vitro fermentation assays with piglet microbiota. Intestinal samples and contents were obtained for small and large intestinal morphology, small intestine disaccharidase activity, and pH and VFA concentrations in ileal (ILE), ascending (AC) and descending colon (DC) contents were measured. Data were analyzed by one-way ANOVA via the MIXED procedure with Tukey-Kramer adjustments for the diet effects. Results: Formula with g-CD alone or g-CD/TB at either dose was well tolerated and did not affect formula intake or piglet growth. ILE morphology was unaffected by supplementation, but there was a numerical trend (p<0.06) for greater DC cuff depth and cuff area in CD-HIGH and CD/TB-HIGH than CON, CD-LOW and CD/TB-LOW. Jejunal sucrase activity was higher (p<0.05) in the CD/TB-LOW than CON, CD-HIGH and CD/TB-HIGH, but it was not different than CD-LOW. ILE pH in CON was significantly higher than any other diet, while AC and DC pH did not differ among the treatment groups. ILE butyrate and propionate concentrations were significantly higher in CD/TB-HIGH than CON, CD-LOW and CD/TB-LOW, but were not different than CD-HIGH. DC acetate concentration was higher in the CD/TB-LOW than CON, CD-HIGH and CD/TB-HIGH, while the difference between CD/TB-LOW and CD-LOW was at a trend level (p<0.08). Conclusion: Encapsulated g-CD/TB complex was well tolerated by healthy piglets. The biological effects of the g-CD/TB complex were dose- and region-specific. On the outcomes herein, the effects of the g-CD/TB complex at each dose was similar to g-CD, alone, supporting the prebiotic potential of g-CD. On-going analyses are determining the effect of g-CD/TB on intestinal microbiota composition and its impact on intestinal health in a piglet model of colon inflammation.
Publications
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2019
Citation:
Shi, X. and Lee, Y. 2019. Encapsulation of tributyrin with whey protein isolate prepared by 2- and 3-fluid nozzle spray drying. IFT Annual Meeting, New Orleans, LA June 2019.
- Type:
Conference Papers and Presentations
Status:
Submitted
Year Published:
2019
Citation:
Marcia H. Monaco, Xueqian Shi, Soo-Yeun Lee, Mei Wang, Claudia Perkowski, Youngsoo Lee and Sharon M. Donovan. 2019. Screening the acceptability and biological effects of encapsulated tributyrin and gamma-cyclodextrin complex on piglet intestinal development. Origins and benefits of biologically active components in human milk conference. West Palm Beach, FL, July 2019.
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Progress 03/15/17 to 03/14/18
Outputs
Target Audience:This project is targeting the scientific community and product developers in the food/nutrition industry who are seeking the solutions to enhance intestinal health by incorporating and delivering butyrate into food matrices. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A graduate student (Ph.D.) and several undergraduate students have been trained during this period. The graduate student has been trained to create TB-CD complex and operate a spray dryer, gas-chromatography (GC), X-ray crystallography (XRD) and scanning electron microscope (SEM). The graduate student analyzed the data and presented the work at a scientific meeting.Also, the undergraduate students were trained to prepare samples from the spray drying. How have the results been disseminated to communities of interest?The results from this part of the project have been presented at a scientific meeting (the Institute of Food Technologists in Las Vegas, Nevada).A symposium session was also organized by PD, and the preliminary work was presented at the IFT meeting. What do you plan to do during the next reporting period to accomplish the goals?The sensory and in vivo tests will be conducted in the next reporting period to examine the bitterness masking efficiency and butyrate release in the GI tract.
Impacts
What was accomplished under these goals?
Butyrate is a short chain fatty acid with a record of improving gastrointestinal (GI) tract health, such as relieving the symptoms of short bowel syndrome, irritable bowel syndrome, and decreasing the proliferation of colon cancer cells in vitro. Tributyrin (TB), a triglyceride with three butyrate molecules, could function as a source of butyrate. However, its extreme bitter taste limits the application. In our previous study, an oven-dried complex of TB with γ-cyclodextrin (CD) has provencapable of masking the bitterness as well as successfully delivering butyrate to the colon. The objective of this period of the project was to improve the production efficiency of TB/CD complex using spray drying. The effect of CD on the solubility of TB and their complexation stoichiometry was studied by a phase-solubility diagram. TB/CD complexes were formed at three different molar ratios, TB: CD of 1:1, 1:2 and 1:3. The complexation rate was evaluated to optimize the complexation time before spray drying. The total and surface TB content was analyzed by gas-chromatography (GC), and the encapsulation efficiency was then calculated. X-ray crystallography (XRD) was used to confirm the formation of the complex. The morphology of the powder was analyzed by Scanning electron microscope (SEM). According to the phase-solubility diagram, the intrinsic aqueous solubility of TB at room temperature was 0.41mM, or 0.127 ± 007 mg/ml. The TB-CD curve can be classified as Bs type, indicating the complex will precipitate after formation. The stoichiometry of TB and CD is between 1:1 and 1:2 (1: 1.3±0.06) indicating the optimum TB:CD ratio would be between 1:1 and 1:2. The complexation will reach 88% after 2 hoursof mixing, which was used as the complexation time before spray-drying. The spray-dried sample with 1:2 molar ratio contains 0.11g/g TB with highest retention (100%) and encapsulation efficiency (97.3%) with 2.7% surface oil. Spray-dried TB/CD showed crystalline structure, supported by both XRD and SEM images. The formation of the TB/CD complex was confirmed by XRD spectra. This study suggested that the production of the TB/CD complex using spray drying is feasible and much more efficient than the previous method.
Publications
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