Source: UNIVERSITY OF CALIFORNIA, DAVIS submitted to NRP
BIOGUIDED FOOD PROCESSING: IDENTIFICATION OF BIOACTIVE INGREDIENTS IN SIDE-STREAMS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0232719
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Oct 1, 2012
Project End Date
Sep 30, 2017
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIVERSITY OF CALIFORNIA, DAVIS
410 MRAK HALL
DAVIS,CA 95616-8671
Performing Department
Food Science and Technology
Non Technical Summary
Agriculture and food need production and processing models that recognize intrinsic biological value of food byproducts and sidestreams, transforming them into new products and methods capable of improving the health of target populations. Commercial viability of large scale extraction of bioactive molecules from agricultural commodities will depend on the richness of the sources. Bovine milk contains only trace amounts of these valuable compounds, however many other agricultural product side-streams are potentially rich in bioactive oligosaccharides. Whey is a major by-product of cheese making, which, for environmental reasons, presents a difficult waste disposal problem. Many important applications await the isolation and characterization of the functional protective components from whey. The importance of this project to the food industry is to provide an industrial proof-of-principle that bioactive components can be extracted from dairy streams in sufficient quantities to perform in-vitro and in-vivo assays. To achieve this goal, the project will use whey streams and membrane technologies combined with modern chromatography and mass spectrometry to characterize the detailed molecular structures of the enriched oligosaccharides and document their efficacy. Bioguided analysis and design of agriculture and food processing side streams can result in the transformation of waste into value-added products and disposal costs into product revenue, while lowering environmental impact, and improving the health of the population. With massive amounts of agricultural byproducts available for capture in California, the methods used to identify, extract, and deliver health-promoting glycans from these streams then becomes invaluable as model for guiding the future of food process engineering, product innovation, and delivery of personalized foods. Results from of this study will provide the foundation for our development of a knowledge base that associates production influences on the relative abundance of bioactivity molecular components. By using mass spectrometry we will gain valuable information about the structures of the bioactive food components so that we can then apply this information to guide separation process from agricultural streams. Unlike most novel food ingredients in which the basic commodities are in low supply and industrial technologies to isolate them unexplored, this project has already assembled the core technologies to characterize the complex oligosaccharides in a widely available substrate. This is also a foundational study, as by characterizing specific bioactive components in dairy streams, it provides preliminary data for subsequent studies looking at specific conditions and study populations. All research findings will be disseminated to the scientific community through publications in peer-reviewed journals and presentations at scientific meetings.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
4033450200035%
5023470202035%
7013470100030%
Knowledge Area
403 - Waste Disposal, Recycling, and Reuse; 502 - New and Improved Food Products; 701 - Nutrient Composition of Food;

Subject Of Investigation
3450 - Milk; 3470 - Other dairy cattle products;

Field Of Science
2020 - Engineering; 1000 - Biochemistry and biophysics; 2000 - Chemistry;
Goals / Objectives
Although traditionally much of the research on prebiotics has used inulin and fructo-oligosaccharides, recent momentum has been behind the glycosylated components found in mammalian milks: oligosaccharides, glycoproteins and glycolipids. Human clinical studies with purified oligosaccharides are lacking, as large amounts of milk oligosaccharides are not available. Goal of this project is evaluating sources of complex carbohydrates other than human milk. Subtle differences in the structures of these molecules can cause significant differences in their prebiotic properties. Therefore, the use of advanced mass spectrometric instruments (High-Accuracy Mass Spectrometry combined with Nanoliquid Chromatography) is crucial to develop comprehensive profile of functional prebiotics in agricultural sources. This research aims at identifying rich sources of oligosaccharides glycoproteins and glycolipids with the biological, structural, glycomic functionalities of human milk containing the key structural elements considered crucial for biological effects such as selective prebiotic activity. As part of this project we propose to develop methods for fractionating gram amounts of the following bioactive components to be tested in functional studies: i) free oligosaccharides ii) glycoproteins, and, iii) glycolipids. Expected outputs: The proposed work includes a first step of enrichment by pilot scale membrane filtration (combination of ultrafiltration and nanofiltration) of dairy streams followed by more specific separations. Using size-exclusion chromatography in combination with activated carbon we expect to fractionate the components based on chemical properties. The high accuracy provided by the analytical strategy proposed (nano-HPLC chip-Time Of Flight mass spectrometer and Gas Chromatography) will enable to successfully validate the new separation methods, achieving unambiguous compound identification that is essential to the successful future application as health promoting ingredients.
Project Methods
A robust analytical approach is necessary to have accurate information about the structure of the bioactive compunds to monitore progress in the isolation and form a coherent basis for interpreting their bioactivities in relation to health. The use of advanced Mass Spectrometry has already allowed the identification of hundreds of oligosaccharides in human milk. This project will add new information by creating bioinformatics libraries that comprehensively characterize the structures of oligosaccharides from different food stream sources. Mass Spectromtric analyses are performed with an Agilent 6520 accurate-mass Quadrupole-Time-of-Flight (Q-TOF) LC/MS with a microfluidic chip (Agilent Technologies, Santa Clara, CA, USA) as previously described (Wu et al., 2011). The microfluidic chip consists of an enrichment column and an analytical column, both packed with porous graphitized carbon, and a nano-electrospray tip. Separation is performed by a binary gradient of solvent A (3 percent ACN, 0.1 percent formic acid (FA) in water) and solvent B (90 percent ACN, 0.1 percent FA in water). Data are acquired in the positive ionization mode with a mass range of m/z 450 to m/z 2500. The Molecular Feature Extractor function of Mass Hunter Qualitative Analysis Version B.04.00 (Agilent Technologies) is used to generate a list of deconvoluted masses. Oligosaccharide compositions are determined from the deconvoluted mass list with an in-house program, Glycan Finder, with a mass error tolerance of about 5 ppm. Distinct Oligosaccharide structures are identified based on accurate mass and retention times compared to previously identified structures. Monosaccharide compositions are further confirmed by MS/MS analysis. Statistical analyses are performed on the mass list using the software Agilent Mass Profiler Professional. Monosaccharides concentrations are determined with absolute quantification by GC. Extracted Oligosaccharides are hydrolyzed to monosaccharides and then derivatized (trimethylsiallilation) to allow the detection and quantification of each monosaccharide. Response factors of each monosaccharide are calculated for each monosaccharide in comparison to the allose standard.

Progress 10/01/12 to 09/30/17

Outputs
Target Audience:Our target audience is the California food industry. As food scientists, we face one of our greatest global challenges: help the agriculture industry feeding a growing world population while, at the same time, reducing our impact on the environment and decreasing the incidence of diet-related chronic diseases. This challenge can only be met by building a more comprehensive understanding of food through research and then bringing this knowledge to practice through innovation. Yet, many of the most promising solutions to our grand food and health challenges are found at the micro level. In particular, regarding human health, much attention today focuses on the functional ability of some food components to modulate the composition of the human gut microbiota. Oligosaccharides, peptides and glycoproteins represent a new opportunity to prevent or ameliorate major infectious and chronic disease worldwide via manipulation of the human gut microbiota. Our research also helps valorize agricultural by-products from the dairy & produce industries by extracting microbiotamodulating bioactive compounds. This research is directly linked to the largest agricultural industry in California, the dairy industry, with the goal of better converting current waste streams to high value products. The California dairy industry produces over 100 million pounds of dairy streams (cheese whey) each day. Currently, California dairy producers must manage the cost of whey permeate as waste, yet I recently discovered that it contains oligosaccharides structurally far more similar to human milk oligosaccharide than currently available prebiotics. Results from this research suggest that concentrating certain oligosaccharides from whey permeate can be a cost-effective process for the valorization of whey permeate into high quality, profitable novel dairy ingredients. Implementing a systematic recovery of milk glycosylated components from whey permeate would enable cheese makers to capture the value from this by-product, generating high-value ingredients and direct revenue. The industry has recognized the importance of my research by supporting multiple projects. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?7 undergraduates, 1 PhD student, and one visiting professor were trained on our methods. How have the results been disseminated to communities of interest?Over this reporting period I have published over 50 peer-reviewed scientific papers and addressed the industry both in small and large meetings, and have presented my work at many National and International conferences attended by industry participants. Overall, I have shared my work at over 50 events open to industry. My commitment to disseminate knowledge to key stakeholders is shown through repeated participation at the Annual Dairy Ingredients Symposium, the IFT annual meeting, the CIFAR Technology Forum and the International Milk Genomic Consortium, the 10th Vahouny Dietary Fiber Symposium, Probiotas Americas, Natural Products Expo and FASEB Science Research Conferences. Invited presentations at international conferences included: FAPESP-UC Davis Research Symposium in São Paulo (Brazil), Foodomics (Cesena, Italy), the John E. Kinsella Memorial Symposium and Lecture, University College Dublin (Ireland), the 4th International Conference on Food Digestion, (Naples, Italy) as well as invited lectures at the Future Food Institute of the University of Modena and Reggio Emilia, Italy, Food Innovation Summer School Mediterraneo, University of Messina (Sicily, Italy). Other outreach activities include speaking at the Davis Rotary Club, California State University Chico, California State University Stanislaus and the Dairy Innovation Institute at Calpoly (San Louis Obispo). I have been actively engaged with CA key stakeholders in filling questions by email and by telephone and sharing recently acquired knowledge regarding bioactive components isolation and recovery. During the review period I engaged with nearly 60 industries. Furthermore, I participated in over 37 interviews during this review period, with various media outlets including PBS (Public Broadcasting Service), The New Yorker, Discovery Magazine, CE&N, Science News, Scientific American, and the Washington Post. Overall, our research targets industry challenges and will provide innovative solutions to enable more readily available supplies of bioactive ingredients. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Key goals for this AES project are improving health and sustainability in food processing through identification of bioactive ingredients in foods and side streams --work that is done both in vitro and in vitro in collaboration with a variety of California stakeholders. I deeply value my position in the Agriculture Extension Station and fully enjoy connecting with industry and community stakeholders to co-develop novel solutions for health and sustainability challenges. The project has thus far successfully identified key candidate bioactive molecules (oligosaccharides, glycolipids, and peptides) in food commodities and their agricultural side-streams --notably from dairy, as previously mentioned. Most recently, I also expanded to researching bioactive compounds in wine and coffee, and I am affiliate faculty member in the new UC Davis Coffee Center. Our results demonstrated that concentrating certain oligosaccharides from whey permeate can be a cost-effective process for the valorization of whey permeate into high quality, profitable novel dairy ingredients. Our lab has employed a combination of mass spectrometry and membrane filtration and achieved the isolation of purified glycans from whey in gram to kilogram amounts, thus generating individual standards that were used to facilitate structural characterization of the novel structures and have been used for animal studies. Implementing a systematic recovery of milk glycosylated components from whey permeate would enable cheese makers to capture the value from this by-product, generating high-value ingredients and direct revenue. Global whey production from cheese and casein manufacture amounts to nearly 240 million metric tons with the EU and USA producing nearly 70% of global whey. In the US alone, production of whey protein concentrate (WPC) and isolates (WPI) totaled approximately 494 million pounds in 2015. The increase in the production of high protein dairy ingredients from whey has resulted in an increase in the volumes of the side-product streams being generated. One challenge to fully capturing the "glycan potential" of bovine milk is that much more glycan is bound as glycoproteins and glycolipids and therefore is inaccessible. With our UC Davis collaborator, Dr. Mills, recently identified a novel enzyme from food-grade microbe B. infantis (a common probiotic) that readily cleaves N-glycans from milk proteins. This discovery is exciting and suggests that application of novel purification methods from bovine milk promises to dramatically increase the level of bioactive glycans that can be obtained from existing dairy streams. Additionally, still in collaboration with Dr. Mills' Lab, my lab demonstrated that several of the fractions have strong prebiotic/antibacterial actions against a wide array of pathogens. These lines of research set the stage to enhance and improve existing prebiotics and antimicrobial peptides. I have been working with the Technology Transfer office to establish a dossier of intellectual property around these health-promoting compounds (U.S. Patent No. 61/363,432 and several other pending applications). My aim is to make all of the research that my lab conducts accessible to the industry by proving-out mechanistic discovery with technological scalability. In addition to expanding our repertoire of food and biological sample processing for oligosaccharide and peptide discovery, another line of research investigated structural characteristics and abundance of bioactive glycolipids in dairy streams. We have demonstrated that glycolipids extracted from buttermilk (a side stream of butter production) support selective growth of B. infantis. This research could also find application in the treatment of inflammatory bowel diseases and Crohn's disease by restoring the lipid rafts of the intestinal mucosa and decreasing inflammation. The results were presented at the American Society for Mass Spectrometry 60th Conference and 2 manuscripts were published. Additionally, we have expanded research to plant products. My lab has discovered ways to naturally increase the concentration of glycosylated mannoproteins in wine by examining their release from several non-Saccharomyces wine strains (2 papers were publishing describing the mechanisms of formation/release). I also obtained funding from the coffee industry (Keurig Green Mountain) to investigate the formation and structural modification of bioactive oligosaccharides during roasting and brewing and their presence in spent grains (1 manuscript published). These coffee oligosaccharides display a wide range of size and monosaccharides, suggesting the potential to selectively stimulate the growth of beneficial bacteria. We also investigated apple pomace, an abundant CA industrial by-product, and found that is rich in dietary fiber and therefore can be a source for bioactive nondigestible oligosaccharides. A project sponsored by Nutrilite allowed us to explore the potential of water-soluble oligosaccharides in plant industrial residues after the desired nutrients have been extracted by the Nutraceutical industry. For example, citrus is an important crop for CA and is mainly used to produce juice, resulting in the accumulation of high amounts of by-products such as citrus peels and seeds. Similarly, dandelion, nettle root, chia seeds, watercress, parsley, Echinacea and sting nettle root have been extensively studied for their physiological and pharmacological effects, however the prebiotic potential of their fiber-rich waste steams had been so far neglected. Our work revealed the presence of diverse and complex oligosaccharides in by-products of all the all of the above listed plant materials (one paper in preparation for publication). The work on these plant-based processing residues has permitted my group to abstract the methods developed for milk, and begin rendering them for universal profiling and quantifying oligosaccharides in all agricultural products.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: May 8, Panel "Innovations for nutrition & sensory quality of foods: Ag Innovation Showcase hosted by The World Food Center". The audience included entrepreneurs, investors in ag/food tech companies, along with established companies in these industries, with a focus on high value crops and livestock.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: June 8, Invited key note: "The effect of dairy products on obtaining a healthy intestinal microbiota", 44th Nordic Dairy Congress,Copenhagen (Denmark), 7-9 June 2017..
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: June 12, An update on milk functional glycomics, Arla Foods Ingredients Group, R&D center, S�nderupvej 26, 6920 Videb�k, Denmark.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: June 13, Application of Glycoproteomics and peptidomics to food product development, Arla Foods Ingredients Group, R&D center, S�nderupvej 26, 6920 Videb�k, Denmark.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: September 20, Invited talk "Oligosaccharides from Whey: Where Are We & What is Needed to Move Forward?" in the Emerging Ingredients Session of the 2017 International Whey Conference Chicago, September 17-20.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: September 21, Invited presentation "The power of whey: old and new activities". Land O'Lakes R&D and Marketing teams, Arden Hills, MN.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Cohen JL., Barile D., Liu Y., de Moura Bell JMLN. Role of pH in the recovery of oligosaccharides from dairy streams by nanofiltration. International Dairy Journal, 66: 6875.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Sischo W., Short D., Geissler M., Bunyatratchata A., Barile D. Comparative composition, diversity, and abundance of oligosaccharides in early lactation milk from commercial dairy and beef cows. 100: 110.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Boudry G. Hamilton M.K., Chichlowski M., Wickramasinghe S., Barile D., Kalanetra K.M., Mills D.A., Raybould H. E. Bovine milk oligosaccharides decrease gut permeability and improve inflammation and microbial dysbiosis in diet-induced obese mice. Journal of Dairy Science, 100: 1-11.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Le Parc A., Karav S., Rouqui� C., Maga E.A., Bunyatratchata A., Barile D. Characterization of recombinant human lactoferrin N-glycans expressed in the milk of transgenic cows. PlosOne, | DOI:10.1371(February 7, 2017): 1-15.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Hamilton, M., Ronveaux, C., Rust, B., Newman, J., Hawley, M., Barile, D., Mills, D., and Raybould, H. Prebiotic milk oligosaccharides prevent development of obese phenotype, impairment of gut permeability and microbial dysbiosis in high-fat fed mice. American Journal of Physiology-Gastrointestinal and Liver Physiology, 1(312(5)): 474-487.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Tian, T., Freeman, S., Corey, M., German, J.B., Barile, D. Chemical Characterization of Potentially Prebiotic Oligosaccharides in Brewed Coffee and Spent Coffee Grounds. Journal of Agricultural and Food Chemistry, 65(13): 27842792.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Mart�n-Ortiz, A., Barile, D., Salcedo, J., Moreno, F.J., Clemente, A., Ruiz-Matute, A.I., Sanz, M.L. Changes in caprine milk oligosaccharides at different lactation stages analyzed by high-performance liquid chromatography coupled to mass spectrometry. Journal of Agricultural and Food Chemistry, 65(17): 35233531.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Smilowitz, J.T., Lemay, D.G., Kalanetra, K.M., Chin, E.L., Zivkovic, A.M., Breck, M.A., German, J.B., Mills, D.A., Slupsky, C., Barile, D. Tolerability and safety for the intake of bovine milk oligosaccharides extracted from cheese whey in healthy human adults. Journal of Nutritional Science, 6(doi:10.1017/jns.2017): 1-11.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Karav S, German J.B.,Rouqui� C., Le Parc A., Barile D. Studying Lactoferrin N-Glycosylation. International Journal of Molecular Sciences, 18(4): 870.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: de Moura Bell J.L.N., Cohen J., de Aquino L. F. M., Lee H., de Melo Silva V.L., Liu Y, Domizio P., Barile D. An integrated bioprocess to recover bovine milk oligosaccharides from colostrum whey permeate. Journal of Food Engineering, 216: 27-35.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: de Aquino L. F. M., de Moura Bell J.L.N.,C., Cohen J., Liu Y, Lee H., de Melo Silva V.L., Domizio P., Conte C.C Junior, Barile D. Purification of Caprine Oligosaccharides at Pilot-scale. Journal of Food Engineering, 214: 226-235.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: OSullivan A., Henrick B.M, German JB., Zivkovic A., Smilowitz J., Barile D., Martin W., Dixon B, Schaefer SE . 21st Century Toolkit for Optimizing Population Health through Precision Nutrition. Critical Reviews in Food Science and Nutrition, 1: 1-12.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Murray N.M., ORiordan D., Jacquier J.C, OSullivan M., Cohen J., Heymann H., Barile D., Dallas D.C. Validation of a paper-disk approach to facilitate the sensory evaluation of bitterness in dairy protein hydrolysates from a newly developed food-grade fractionation system. Journal of Sensory Studies., 32(3): e12266.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Tian T. and Barile D. The Future of Coffee and Its Waste Streams: Novel Health Benefits Ahead. The World of Food Ingredients.February 2017 ISSN 1566-6611, pp 62-65.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Shimoda M., Galermo A.G., Olney L., Barile D., German B., Lebrilla C., Maverakis E. 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. Modulation of dendritic cell differentiation and function by human milk oligosaccharides.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Weinborn V., Li Y., Chen X., Barile D. 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. Enzymatic modification of bovine milk oligosaccharides and their functional properties. Student Travel Award Recipient (poster selected for oral presentation).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Bunyatratchata A., Karav S., Frese S.A., Shaw C.A., Contreras L.N., Mills D.A., Barile D. 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. Sialic acid release by Bacteroides thetaiotamicron from milk oligosaccharides may facilitate the growth of potentially pathogenic bacteria. Student Travel Award Recipient (poster selected for oral presentation).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Robinson R.C., Poulsen N.A., Larsen L.B., Barile D. 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. A novel method for high-throughput analysis of bovine milk oligosaccharides. Student Travel Award Recipient (poster selected for oral presentation).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Robinson R.C., Poulsen N.A., Larsen L.B., Barile D. 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. Discovering natural bioactive peptides in cheese with mass spectrometry. Student Travel Award Recipient (poster selected for oral presentation).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Cohen J.L., Karav S., de Moura Bell J.M.L.N., Mills D.A., Barile D. 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. Pilot scale isolation of bioactive glycans from dairy co-products: Capturing the whey glycome. Student Travel Award Recipient (poster selected for oral presentation).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Salcedo J., Goonatileke E., Guerrero A., Kimzey M., Haxo T., Lebrilla C., Barile D. 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. Published in the Journal Glycobiology 26 (12) pp 1469-1469. High-throughput milk oligosaccharide analysis using a rapid cartridge-based capillary electrophoresis instrument.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Salcedo J., Frese S.A., Mudd A., Dilger R.N., Chichlowski M., Berg B.M., Mills D.A., Barile D. 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. Glycosylated bioactives compounds in porcine milk: identification and quantification during lactation and correlation with the fecal metagenome.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Shah I.M., Frese S.A., Xu G., Lebrilla C.B., De Moura Bell H.M.L.N., Barile D., Mills D.A. 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. Cross-feeding of pathobionts by Bacteroides thetaiotaomicron as a possible mechanism for Necrotizing Enterocolitis.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Shani G., Davis J., Cohen J., Nagshbandi S., Ho S., Ruiz-Moyano S., Lewis Z, Popovic M., Frese S., DeMoura Bell J., Barile D., 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. Diversity and Milk Oligosaccharide Consumption of Bifidobacterium pseudocatenulatum.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Barile D., Salcedo J., Le Parc A., Sun A., Karav S., Cohen J., DeMoura Bell J. 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016. KEYNOTE: Milk glycoproteomics: Preserving, enhancing, and delivering bioactivity.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Mart�n A., Salcedo J., Barile D., Clemente A., Moreno F.J., Ruiz A.I., Sanz M.L. XVI Annual Meeting of the Spanish Society of Chromatography and Related Techniques (SECyTA2016). 2-4 November, Seville, Spain. Comprehensive Analysis of Goat Milk Oligosaccharides.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Tian T., Barile D. IFT16, July 2016, Chicago. Identification and Characterization of Prebiotic Oligosaccharides in Plant material.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Shankar S., Barile D., Lange M. Ontological framework for bioactive compounds in milk and dairy products. Poster presented at the First International Conference for Food Ontology, Operability, Data and Semantics. UC Davis November 7-9.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Duchene C., Laborie E., Barile D., Lange M. Poster presented at the First International Conference for Food Ontology, Operability, Data and Semantics. UC Davis November 7-9. 2016. Milk Mapper: A mammalian milk oligosaccharides database enhancing unique knowledge and advances in human health.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Salcedo J., Frese S.A., Mills D.A., Barile D. FST RMI Symposium December 8, UC Davis . Glycosylated bioactives compounds in porcine milk: identification and quantification during lactation and correlation with the fecal metagenome.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Gan J., Robinson R., Barile D., German J.B. FST RMI Symposium December 8, UC Davis. pH-Specific Proteolysis of Human Milk: Biological Implications for Infants.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Nocerino R., Picariello G., Ferranti P., Paparo L., Passariello A., Dallas D.C., Robinson R.C., Barile D., Canani B.R. In: Allergy 2016 Aug 1 (Vol. 71, pp. 18-18). 111 River St., Hoboken 07030-5774, NJ USA: Wiley-Blackwell. Antibody-independent detection of bovine beta-lactoglobulin derived peptides in human milk.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Westreich S.T., Barile D., Mills D.A., Smilowitz J.T., Korf J., Lemay D.G. Experimental Biology 2017, Chicago April 22-26. Using Metatranscriptomics to Determine Effects of Dietary Supplementation with Bovine Milk Oligosaccharides in Healthy Adults. Also published in The FASEB Journal 31 (1 Supplement), 650.30-650.30, 2017.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Frese S.A., Smilowitz J.T., Hutton A.A., Contreras L.N., Shaw C.A., Palumbo M.C., Xu G., Davis J., Lebrilla C.B., Freeman S., Barile D., German J.B., Mills D.A., Underwood M. 2017 Pediatric Academic Societies Meeting, San Francisco CA, May 6-9, 2017. Loss of a key symbiont has changed fecal biochemistry in breastfed infants.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Henrick B.M., Erhlich A.E., Vang W., Cohen J., German J.B., Mills D.A., Barile D., Raybould H.E. Milk oligosaccharides decrease pathogen associated molecular pattern induced inflammation in intestinal epithelial cells. FASEB Journal. 31(1): Supplement 315.3.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Bhattacharya M., Robinson R., Barile.D. IFT Annual Conference, Las Vegas, NV, June 25 - 28, 2017. Peptidomic profiling of commercial dairy products: Identification, functional analysis and effect of processing .
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Cowardin C.A., Ahern P.P., Guruge J., Barratt M., Barile D., Mills D.A., Gordon J.I. International Congress of Mucosal Immunology 19-22 July 2017. Sialylated milk oligosaccharides promote microbiota-dependent immune and skeletal development in a model of infant undernutrition.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Gan J, Barile D, Lebrilla CB., German JB. 14th International Symposium on Milk Genomics & Human Health, September 26-28, 2017 Qu�bec City, Canada.. Milk as a protein-protease delivery system: collaborative approaches inspire inquiry into the dynamics and complexities of human milk.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Smilowitz J.T., Frese S. A., Hutton A.A., Contreras L.N., Shaw C.A., Lebrilla C.B., Freeman S. L., Barile D., German J.B., Mills D.A., Underwood M.A. American Academy of Pediatrics Annual Conference, September 16-19, 2017, Chicago IL. Bifidobacterium longum subsp. infantis stably restores the infant gut microbiome over the first year of life in breastfed infants. .
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Poulsen N.A., Buitenhuis B.., Robinson R.C., Barile D. , Larsen L.B. International Milk Genomics Consortium (IMGC) annual conference. September 26-28, Qu�bec City, Canada. Genetic parameter estimation for bovine milk oligosaccharides in Danish dairy breeds.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Robinson R.C., Poulsen N.A., Larsen L.B., Barile D. International Milk Genomics Consortium (IMGC) annual conference. September 26-28, Qu�bec City, Canada. Selectd for Travel Award. Comparative analysis of bioactive oligosaccharide production in dairy cows using novel analytical techniques.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: November 8, Invited talk: "Challenges in Processing Bioactive Food Components". IC-FOODS ( International Conference/Consortium/Center for Food Ontology, Operability, Data and Semantics) Nov. 7-9, 2016 UC Davis Conference Center.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: November 18, Invited talk at the Chemistry Careers Club, Department of Chemistry Colloquium. California State University Stanislaus.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: November 23, Invited lecture: Milk as a model for food digestion.The Food Innovation Masters Program 2.0. University of Modena and Reggio Emilia, Italy.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: November 24, Invited talk "The Food of the Future". Food Is A Conversation with Daniela Barile. Food Innovation Program, event open to the city of Reggio Emilia, IT.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: November 25, Invited lecture: Innovating Dairy Ingredients with a multi-omics approach.The Food Innovation Masters Program 2.0. University of Modena and Reggio Emilia and the Young Talents Academy, Italy.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: November 26, Invited lecture: "Comparing the Italian and American dairy system: valorization of by-products". The Food Innovation Masters Program 2.0. University of Modena and Reggio Emilia, Italy.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: December 6, Invited talk " Technological Innovation in the Agro-Food sector". Istituto di Cultura Italiana (Institute of Italian Culture), San Francisco. A World-Expo follow-up event organized by the Italian Minister of Agriculture.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: December 8, Invited talk: "Health benefits of bioactive compounds extracted from agricultural side-streams". FST Research Symposium, UC Davis.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: January 11, Invited talk "Harvesting Milk Components for Health". Annual meeting of the California Creamery Operators Association. UC Davis.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: February 14, Invited presentation "Feeding the world in the XXI Century". World Ag Expo; research track Hosted by College of Agricultural and Environmental Sciences, UC Davis at the International Agri-Center in Tulare, CA .
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: March 20, Tour with presentation "Innovations in Milk Research from the Milk Processing Lab" UC-wide Research Development (UCRD) conference. This meeting involved individuals from the 10 UC Campuses teaming up for strategic initiatives and interdisciplinary research.


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Our target audience is the California food industry. As food scientists, we face one of our greatest global challenges: help the agriculture industry feeding a growing world population while, at the same time, reducing our impact on the environment and decreasing the incidence of diet-related chronic diseases. This grand challenge can only be met by building a more comprehensive understanding of food through research and then bringing this knowledge to practice through innovation. Yet, many of the most promising solutions to our grand food and health challenges are found at the micro level. In particular, regarding human health, much attention today focuses on the functional ability of some food components to modulate the composition of the human gut microbiota. Cataloging and quantifying the varied biological roles and functions of these bioactive food components is both urgent and important. In contrast to the science of essential nutrients, the nascent field of research around bioactive molecules and their capacity to modulate health altering biological processes requires novel tools and methods to advance. These molecules act on many different physiological systems through a wide variety of mechanisms, ranging from interaction with commensal bacteria to binding viruses and toxins. To realize health in this new millennium, we must deepen our understanding about this class of molecules: their sources in foods, their functions and response to processing, and concomitant impact on human health. Nutrients are defined as molecules that are essential for human health. Bioactive molecules have not been shown to be essential for human health, but have the capacity to modulate biological processes that enhance health. Many non-nutrient, bioactive constituents are being investigated and annotated for positive health benefits. These molecules act on many different physiological systems through a wide variety of mechanisms, ranging from interaction with commensal bacteria to binding viruses and toxins. The intestinal microbiota has become a recent target of research because of its importance as a health factor for infants and adults. As new analytical technologies are assembled, new functional milk molecules are being discovered in foods and agricultural streams. Innovative research by the Barile Lab has shown that bioactive molecules from agro-food chains can be captured and valorized, creating new opportunities for promoting human health while also reducing waste. Oligosaccharides, peptides and glycoproteins represent a new opportunity to prevent or ameliorate major infectious and chronic disease worldwide. Our research helps valorize agricultural by-products from the dairy & produce industries. This research is directly linked to the largest agricultural industry in California, the dairy industry, with the goal of better converting current waste streams to high value products. The California dairy industry produces over 100 million pounds of dairy streams (cheese whey) each day. Currently, California dairy producers must manage the cost of whey permeate as waste, yet I recently discovered that it contains oligosaccharides structurally far more similar to human milk oligosaccharide than currently available prebiotics. Results from this research suggest that concentrating certain oligosaccharides from whey permeate can be a cost-effective process for the valorization of whey permeate into high quality, profitable novel dairy ingredients. Implementing a systematic recovery of milk glycosylated components from whey permeate would enable cheese makers to capture the value from this by-product, generating high-value ingredients and direct revenue. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?My student Tian Tian Presented an e-poster (short-communication) titled: "Identification and Characterization of Potentially Prebiotic Oligosaccharides in Plant material". Tian also attended the following seminars: · "Career Assessment Series". January 6 and 25, 2016. University of California, Davis · "Prepare for the Interview for Teaching Positions in Academia". January 19, 2016. University of California, Davis · Effective Communication for Career Success: Practice Telling Your Story". February 9, 2016. University of California, Davis · "Writing a Literature Review". April 27, 2016. University of California, Davis · Key to Effectively Delivering Scientific Presentations and Posters". October 6th, 2016,, University of California, Davis My student Randall Robinson presented a Poster titled: "A novel method for high-throughput analysis of bioactive oligosaccharides" at the HPLC 2016, June 2016, San Francisco, CA. · Randall also presented twice at the International Milk Genomics Conference (October 2016); Presentation titles: "A novel method for high-throughput analysis of bioactive oligosaccharides" and "Discovering natural bioactive peptides in cheese with mass spectrometry". · Randall also attended the following seminars: · Made to Stick: Keys to Effectively Delivering Scientific Presentations and Posters (GradPathways, Oct 6, 2016) · Understanding Learning Styles for Leadership Development (GradPathways, Oct 19, 2016) · Agilent LC/MS Preventative Maintenance workshop (Nov 12, 2015) · Agilent Breaking Bad Chromatography Habits (Mar 26, 2015) How have the results been disseminated to communities of interest?I am committed to translating and communicating scientific knowledge to fellow scientists and the general public. In 2015 have addressed the industry both in small and large meetings, and have presented my work at many National and International conferences attended by industry participants including the "Emerging Markets, Technologies and Products Using Milk and Dairy Ingredients" at the 18th Annual Dairy Ingredients Symposium (February 17-18, 2016 Shell Beach, CA), the "Probiotic Educational Track" at Natural Products Expo (March 9, 2016 Anaheim Convention Center, CA), the "Human Milk Oligosaccharides for Environmental Enteric Dysfunction Workshop" at the Bill & Melinda Gates Foundation (May 17, 2016 Seattle, WA), "Prebiotics - Quantifying impact on host health' workshop", IPA World Congress, Probiotas Americas (May 31, 2016 Chicago), the 13th International Symposium on Milk Genomics and Human Health, (27-29 September 2016 Davis CA) and more (see complete list under "Products"). I have actively engaged key stakeholders and the CA dairy industry in sharing our recently acquired knowledge regarding bioactive components in food and technologies for isolation and recovery of compounds from food matrices. I have been also actively engaged in outreach to the broader public. These activities include my interviews with PBS, The New Yorker and other media outlets including an international documentary on food. During the reporting period I presented my research to the following food industries: Leprino Foods, Mars Inc., Land O'Lakes, Sterling Technology, Mead Johnson, Volac International, Mengniu, Ylli, 4LIfeResearch, Arla Amba, Glanbia, Joseph-Gallo Farms (CA), LaBelle (CA), and, Hilmar Ingredients (CA). What do you plan to do during the next reporting period to accomplish the goals?Currently, little is known about factors that impact bovine milk oligosaccharides concentration in milk even though variation is known to occur. Understanding the reasons for these natural variations would allow the environmental factors (e.g. pasture feeding) to be harnessed so that milk can be tailored to fit its eventual application. By establishing the abundances of free oligosaccharides in bovine milk from various feeding trials (e.g. pasture fed versus industrial settings), we could provide a guide for the dairy industry to strategically capture the most important components with regard to glycosylation. This approach will eventually enable development of new dairy products with health guiding capabilities for infants, immuno-compromised elderly, and the population at large. Additionally, I plan to continue expanding my network of collaborators and explore other suitable environmentally friendly processing technologies for the recovery of large-scale quantities of oligosaccharides adding value to key food streams relevant to California agriculture such as grape pomace, brewery spent grains, coffee grounds, olive & apple pomace, tree nuts skins and shells. Through the use of novel enzymes, I will further facilitate large-scale release of previously unavailable bioactive glycans, and, in collaboration with organic chemists I plan to expand the repertoire of plant-derived carbohydrates using a one-pot multi-enzyme approach decorate simpler glycans increasing their prebiotic efficacy.

Impacts
What was accomplished under these goals? This project focuses on identification of bioactive ingredients in food side streams and addresses the key goals of improving health and achieving sustainability in food processing. During this reporting period my group published 11 peer-reviewed journal articles as well as 1 article in a specialist journal that focuses on product development and nutrition advances affecting the industry (Karav S. & Barile D . Unlocking novel bioactive compounds in milk. The world of food ingredients. 2016 April-May issue, pp 72-77). I also presented this research at 11 national and international conferences (see "Products" for a complete list). This work garnered significant media attention. The industry has recognized the importance of my research by supporting multiple projects on free oligosaccharides, glycoproteins and peptides. I continually contribute to dissemination of knowledge and provide details on new technologies for bioactive compounds extraction and application and their potential impact on the California Dairy Industry and human health. The main goal of this project is evaluating sources of complex carbohydrates that mimic human milk biological, structural, glycomic functionalities and contain the key structural elements considered crucial for biological effects, such as selective prebiotic activity. Despite the important role of human milk oligosaccharides (HMO), these compounds are not well represented in infant formula which is currently supplemented with plant derived oligosaccharides such as fructoligosaccharides or lactose-derived oligosaccharides such as galactooligosaccharides. Currently available commercial oligosaccharides are already used in a wide range of food products, but despite having a general prebiotic effect, they lack many of the other beneficial effects provided by HMO. This difference is mainly caused by the simpler chemical structure and lack of linkages diversity. This work successfully identified key candidate molecules (oligosaccharides, glycoproteins, and peptides) as well as performed their high-throughput characterization and quantification by mass spectrometry and other advanced analytical techniques. These lines of research set the stage to enhance and improve existing prebiotics by translating the evolutionarily-linked beneficial association between milk oligosaccharides and their associated beneficial bifidobacteria. Glycomics and Peptidomics are recent technologies that are allowing food scientists to develop a detailed molecular-level understanding of such functional compounds in milk and other foods. Understanding human and bovine milk from this perspective, and how milk components interacts with different biological processes will enable the development of new food products to guide health in infants, immuno-compromised elderly, and the population at large. This research combines a unique set of collaborations, employing clinical subjects, mass spectrometry and computational algorithms to identify bioactive compounds in foods and food streams. In collaboration with Dr. David Mills' lab, we demonstrated that a novel endo-β-N-acetylglucosaminidase (EndoBI-1) isolated from B. infantis is able to release a wide variety of glycan from proteins. Conversely from existing chemical deglycosylation strategies, which require harsh chemicals or high heat-induced thermal denaturation of the substrate, (which would alter the native form of the polypeptide and possibly damage glycans' most delicate structures), the novel enzymatic approach the ability to cleave N-glycans from native bovine milk proteins and yet it is heat-stable so it can be incorporated directly into conventional dairy processing. This phenomenon could be explained by the fact that EndoBI-1 is isolated from a bacterial specie found in infant gut, and the infant gut microbiota is already active on native glycoproteins found in mother's milk. Hence, the observed higher activity of EndoBI-1 for native whey protein is an expected outcome of its' natural origin. Successful release of glycans on their native form will enable the investigations of this new class of compounds biological roles and their use in food industry. Preliminary results have already shown that glycans released from whey proteins can promote the growth of an important infant human microbiome member (one paper published). The ability to use novel processing enzymes to harvest bioactive HMO-mimics (glycans from proteins) using an abundant and inexpensive substrate such as cheese whey, will enable development of new food products with health guiding capabilities for infants, immuno-compromised elderly, and the population at large as well as add value to an existing agricultural co-product. Another novel aspect of this project was expanding our investigation of oligosaccharides presence from commercial bovine milk to calf feeds and how this relates to indicators of calf health such as the presence of probiotic bacteria. This project, in collaboration with Washington State University, directly addresses the goals of the food safety challenge by developing approaches that reduce unnecessary antibiotic use that lower on farm abundance and diversity of resistant organisms and mitigating the consequences of antibiotic use. There is considerable variability in the quality of milk and replacers and understanding and remediating this variability is a key objective since variability in calf liquid feed affects the occurrence of calf diarrhea and the intestinal microbiota which plays a role in neonatal disease.Because subtle differences in the structures and concentration of these oligosaccharides can cause significant differences in their prebiotic properties, we set to explore this unique environmental influence. To accomplish this, we have quantified total oligosaccharides diversity and abundance in different colostrum and feeding samples using our lab's analytical platform. A set of 15 samples representative of calves's feeding practices was analyzed for presence of bioactive oligosaccharides. Of the 15 samples, 4 were bovine colostrum samples and 11 samples were either bulk milk, milk labelled as hospital milk, milk typically provided to older calves, fresh milk or their combinations. A total of 18 oligosaccharides were identified in the samples other than colostrum samples while 14 oligosaccharides were identified in the bovine colostrum samples. The colostrum samples had a much higher abundance of oligosaccharides by an order of magnitude of 10-100 higher than the non-colostrum samples. In the non-colostrum samples, the mixture of pasteurized fresh milk and milk typically provided to older calves had the highest abundance of oligosaccharides. After having identified important differences in OS composition among colostrum and the various milk replacer, a subsequent aim of this study is to determine whether OS supplementation of colostrum and milk impacts measures of animal productivity. The study outcome will be disease treatment with a focus on diarrhea. This novel study will accelerate translation of a protective mechanism observed in human milk into nutritional strategies to limit intestinal dysbiosis in calves. Our lab also examined the naturally occurring peptides in bovine milk. This work demonstrates that degradation of milk proteins within the mammary gland is remarkably consistent across cows --the same peptides are released from the same proteins, while many abundant proteins are left untouched. Moreover, we whey permeate (the examined an industrial by-product of protein concentration), and revealed that this waste stream contains the vast majority of peptides that naturally occur in bovine milk. We have now successfully extracted the peptides and fractionated them on a pilot-scale. In collaboration with Dr. Mills' Lab, we demonstrated that several of the fractions have strong antibacterial actions against a wide array of pathogens.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: March 9, Invited talk "Next generation prebiotics: challenges and opportunities". Probiotic Educational Track at Natural Products Expo, Anaheim Convention Center.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: April 13, Invited talk: Dairy Streams: challenges and opportunities, ETH Zurich Meets California, Tackling Food System Challenges with IT Innovation, event hosted by the "Mixing Bowl Hub"; Robert Mondavi Institute, Davis CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: May 11, Invited talk: Glycoproteomics investigation of milk, Chancellor/Regent Associates Visit, UC Davis Chancellor's Residence, Davis.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: May 17, Invited talk: Overview of human milk oligosaccharides measurement and capture from animal milks, Human Milk Oligosaccharides for Environmental Enteric Dysfunction Workshop, The Bill & Melinda Gates Foundation | Seattle, WA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: May 31, Invited talk: Prebiotics for premature infants: discovery and characterization, Prebiotics - Quantifying impact on host health workshop, IPA World Congress, Probiotas Americas, Chicago (USA).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: July 16, Invited talk: "Creation of novel foods with an unusual nutritional value", Food Innovation Summer School Mediterraneo, University of Messina, Sicily IT.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: September 24, Invited talk "Characterization and recovery of compounds from natural sources". Society of Environmental Journalists annual conference, UC Davis.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: September 28, Invited Keynote Presentation: Milk glycoproteomics: Preserving, enhancing, and delivering bioactivity, 13th International Symposium on Milk Genomics and Human Health, Davis CA, 27-29 September 2016.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Dallas, D.C., Sanctuary, M.R., Qu, Y., Khajavi, S.H., Zandt, A.E.V., Dyandra, M., Frese, S.A., Barile, D., and German, J.B. Personalizing Protein Nourishment. Critical Reviews in Food Science and Nutrition.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Charbonneau, M., O'Donnell, D., Blanton, LV., Totten, SM., Davis, J.C.C., Barratt, M.J., Cheng, J., Guruge, J., Talcott, M., Bain, J.R., Muehlbauer, M.J., Ilkayeva, O., Wu, C., Struckmeyer, T., Barile, D., Mangani, C., Jorgensen, J., Fan, Y.M., Maleta, K., Dewey, K.G., Ashorn, P., Newgard, C.P., Lebrilla, C.B., Mills, D.A., Gordon, JI. Sialylated Milk Oligosaccharides Promote Microbiota- Dependent Growth in Models of Infant Undernutrition. Cell, 164(5): 859-871.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: de Moura Bell, JMLN, Aquino, LFMC, Liu, Y, Cohen, JL, Lee, H., de Melo Silva, VL, Rodrigues, MI, Barile, D. Modeling Lactose Hydrolysis for Efficiency and Selectivity Towards the Preservation of Sialyloligosaccharides in Bovine Colostrum Whey Permeate. Journal of Dairy Science, 99(8): 6157-6163.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Karav, S., Le Parc, A., de Moura Bell, J.M.N.L., Frese, S., Kirmiz, N., Block, D.E., Barile, D., Mills, D.A. Oligosaccharides released from milk glycoproteins are selective growth substrates for infant-borne bifidobacteria. Applied and environmental microbiology, 82 (12): 3622-3630.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Picariello, G., Addeo, F., Ferranti, P., Nocerino, R., Paparo, L., Passariello, A., Robinson, R.C., Barile, D., Dallas, D.C., Canani, R. B. Antibody-independent identification of bovine milk-derived peptides in breast-milk. Food & Function, 7 (8): 3402-3409.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Karav S., Cohen J.L., Barile D., de Moura Bell J.M.L.N. Recent Advances in Immobilization Strategies for Glycosidases. Biotechnology Progress.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Bode L., Contractor N., Barile D., Pohl N., Boons G.J., Jin Y.S., Jennewein S. Overcoming the limited availability of human milk oligosaccharides: challenges and opportunities for research and application. Nutrition Reviews, 74(10): 635-644.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Lee, H.; Cuthbertson, D.; Otter, D.; Barile, D. Rapid Screening of Bovine Milk Oligosaccharides in a Whey Permeate Product and Domestic Animal Milks by Accurate Mass Database and Tandem Mass Spectral Library. Journal of Agricultural and Food Chemistry, 64(32): 6364-6374.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Domizio P., Liu Y., Bisson L., Barile D . Cell wall polysaccharides released during the alcoholic fermentation by Schizosaccharomyces pombe and S. japonicus: quantification and characterization. Food Microbiology, 61: 136-149.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: A Mart�n-Ortiz, J Salcedo, D. Barile, A Bunyatratchata, FJ Moreno, I Martin-Garc�a, A Clemente, ML Sanz, AI Ruiz-Matute. Characterization of goat colostrum oligosaccharides by nano-liquid chromatography on chip quadrupole time-of-flight mass spectrometry and hydrophilic interaction liquid chromatography-quadrupole mass spectrometry. Journal of Chromatography A, 1428: 143-153.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: DC Dallas, F Citerne, T Tian, VLM Silva, KM Kalanetra, SA Frese, RC Robinson, DA Mills, D. Barile. Peptidomic analysis reveals proteolytic activity of kefir microorganisms on bovine milk proteins. Food Chemistry, 197: 273 284.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: October 26, Moderated industry panel on "Impacts of Microbiomes" at the FALL 2015 University-Industry Consortium Meeting  UC Davis Conference Center.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: November 9, Invited talk: Innovation in Milk Research, Visit of a delegation of Italian Entrepreneurs led by the Governor of Emilia Romagna. UC Davis.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: December 10, invited talk: Beyond essential nutrition: discovering & delivering bioactive compounds, John E. Kinsella Memorial Symposium and Lecture The Fitzgerald Debating Chamber, University College Dublin (Ireland).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: February 17, invited talk: Oligosaccharides Processing from Dairy Fluids  An Update, 18th Annual Dairy Ingredients Symposium: "Emerging Markets, Technologies and Products Using Milk and Dairy Ingredients", February 17-18, Shell Beach, CA.


Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Our target audience is the California food industry. Oligosaccharides, peptides and glycoproteins represent a new opportunity to prevent or ameliorate major infectious and chronic disease worldwide. Our research helps valorize agricultural by-products from the dairy & produce industries. I am committed to translating and communicating scientific knowledge to fellow scientists and the general public. In 2015 have addressed the industry both in small and large meetings, and have presented my work at many National and International conferences attended by industry participants including: the University-Industry Consortium Meeting, FASEB Science Research Conferences, FAPESP-UC Davis Research Symposium in São Paulo (Brazil), Nanjing Agricultural University-UC Davis One Health Food Safety Symposium, theKinsella Memorial Symposium and Lecture in Dublin(Ireland), the World Bank - Haiti Conference on Coffee production, the Food Innovation Program (University of Modena and Reggio Emilia, Italy). I was also an invited speaker at IBM Almaden Research Center's Distinguished Speaker series which was broadcasted in all IMB facilities worldwide. I also participated in several meeting with Mars Inc and the early developments of the UC Davis Innovation Institute for Food and Health and helped shape key research questions that will impact major issues in food, Ag and health. During the reporting period I presented my research to the following food industries: Barilla, Nestle', Novozymes, Indofods Salim Group, EatLimmo, SomaLogic, Friesdland-Campina,Clover Stornetta Farms, Borden Dairy Company, FairLife, Suntory, Rizo Lopez Foods, Clorox, Wrigley (Mars), as well as companies involved in analytical methods development: Shimadzu, Bruker, ThermoFisher Scientific, Prozyme and organizations such as The World Bank and Ag Research. To enable translation of the research to food products, I also worked with local industries such as Hilmar Ingredients (Hilmar, CA), Prolacta Bioscience (Monrovia, CA), LaBelle Inc. (Rippon, CA), Nutrilite (Buena Park, CA). I also initiated projects with the following global corporations: Danone Food Company, Sterling Technology, Arla Foods, Mead Johnson, Volac International, Separation Techniques Limited, Mengniu, Ylli, 4LIfeResearch, Arla Amba, Land O'Lakes and Glambia. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?15 undergraduates, 5 PhD student, and 3 visiting professors were trained on our methods. How have the results been disseminated to communities of interest?A major focus of my project is to engage with industry, academia, and the general public. I have established relationships with large manufacturers of pilot-scale and industrial filtration equipment that are interested in taking our prototype to commercial reality. I visited our collaborators (mostly large dairy companies based in California) and disseminated our research results. I also presented this research to 16 national and international conferences. During this review period, I continued to serve as Associate Editor for the Newsletter "SPLASH!" distributed by the International Milk Genomic Consortium and I contributed 1 piece to educate the industry and the general public in milk research. I have published 2 additional papers in high-impact, editorially reviewed articles read by our stakeholders (food industry, food processors). I was interviewed about my research. What do you plan to do during the next reporting period to accomplish the goals?Further developments include applying our analytical technology and oligosaccharides and N-glycans bioinformatic libraries to explore the potential of plants as sources of bioactive glycans. Successful application of these libraries will allow converting whey, a waste product, into a substrate for selective modulation of the gut microbiota, mimicking what happens in the gut of a healthy breast-fed infant. Additionally, the Product Manager of Thermo-Fisher (Dr. Angrish) visited my lab in October and was very impressed with our research: as a results of this visit she has asked my lab to co-develop an application note for measuring oligosaccharides on the ICS5000. Similarly, Dr. Guerriero, Product Development Specialist of Prozyme, has offered my lab the opportunity of beta-testing a new Capillary-Electrophoresis system that has the potential to be used in glycans rapid monitoring (this project should be starting in February-March 2016). With all these technologies in place, it will soon be possible to assess and identify which foods and food streams yield the highest concentrations of functional glycans to obtain the necessary amounts for functional assays and clinical trials.

Impacts
What was accomplished under these goals? During the past year our work was focused on the optimization of carbohydrates purification, quantification and characterization to obtain purified fractions for functional testing. During this reporting period my group published 18 peer-reviewed journal articles as well as 1 book chapter and 8 abstract. I also presented this research at 16 National and International conferences. Below I will describe in further detail the activities for the project in terms of advances in Glycoproteins, Free oligosaccharides and Milk Processing research.? Glycoproteins Research Our sophisticated analytical platforms are enabling a detailed molecular-level understanding of functional compounds in milk and other foods. Over the last year we evaluated the effects of pH, temperature, reaction time, and enzyme/protein ratio on the total release of N-glycans from food glycoprotein using a new enzyme (EndoBI-1). Denaturation of substrates for deglycosylation is also an essential step for enzymatic release of glycans from glycoproteins; however, when studying functional activities denaturation is highly undesirable as it may mask/compromise some tests. Interestingly, our recently isolated enzyme (EndoBI-1) shows higher activity compared to other glycosidases even in absence of denaturation. We compared the activity of the commercial enzyme PNGase F and our EndoBI-1 (cloned at the lab) and we demonstrated a comparable activity on ribonuclease B (a simple glycoprotein). Importantly, when using a real substrate such as bovine whey and the milk protein lactoferrin, our EndoBI-1 enzyme yielded significantly more N-glycans than PNGase F (a new manuscript was written detailing these findings). Of relevance to the food industry, new enzyme can be efficiently used during pasteurization, streamlining its integration into existing processing strategies. Leveraging our efforts in the purification of free bovine milk oligosaccharides, we achieved further isolation of released glycans enabling, for the first time, in vitro testing of biological activity. These results provide mechanistic support that N-linked glycoproteins from milk serve as selective substrates for the enrichment of infant-borne bifidobacteria capable of carrying out the initial deglycosylation. Moreover, released N-glycans are better growth substrates than the intact milk glycoproteins suggesting that EndoBI-1 cleavage is a key initial step in consumption of glycoproteins. Finally, the variety of N-glycans released from bovine milk glycoproteins suggests they may serve as novel prebiotic substrates with selective properties similar to those of human milk oligosaccharides. Free oligosaccharides Research. Our work expanded from building a library of glycoproteins to free oligosaccharides: to further streamline glycans' identification, our comprehensive free oligosaccharide library was embedded into a custom Personal Compound Database & Library (PCDL) Program. The new library contained 34 individual structures, and included, beside information on accurate mass & retention time, also the full data for each tandem mass spectra - which enables assignment of fine structures with fragmentation pattern arising from unique mass signature. This program was used to rapidly identify unknown oligosaccharide in milk samples by comparing the fragmentation spectra; it resulted to be an efficient and reliable method to identify oligosaccharide structures via automated spectral matching. The rapidly screen Bovine Milk Oligosaccharides (BMOs) in mammalian milks and dairy streams and required only nanogram quantities of material. The program was further applied to screen for other mammalian milks suitable as candidate for oligosaccharides extraction, such as sheep milk and buffalo. Interestingly we demonstrated significant overlap between BMOs and the sheep and buffalo milk oligosaccharides (14 BMOs were found). This easy-to-use and yet comprehensive screening provided great confidence of identification and it will prove a valid tool to investigate glycan structure-function relationships of for milk oligosaccharide research. One manuscript detailing these results was written and it is now in the final stages of review. Another extension of this research line involved collaborating with one of the major coffee roasters (Green Mountain) to investigate the overall distribution of oligosaccharides in brewed coffee and cofffee processing streams. Oligosaccharide with a variety of degree of polymerization (DP), ranging from DP 3 to 16 and their constituent monosaccharides were identified, characterized and quantified. The concentration ranged between 4mg and 12 mg per g of coffee. The oligosaccharides found were mainly composed of hexose oligosaccharides (potentially galacto-oligosaccharides and manno-oligosaccharides) with side chain containing arabinose, xylose and rhamnose. The diversity of monosaccharides composition provided the basis for matching these prebiotics to specific probiotic bacteria. Further assays are being performed to assess the specific of the prebiotic-probiotic interaction. Milk Processing Research. We also continued to expand the laboratory fractionation of oligosaccharides at the pilot-scale.The membrane fractionation methodology selected was similar to the methods commonly used industrially to produce whey protein concentrate and whey permeate. The work included extensive optimization of lactose hydrolysis (lab- and pilot-scale) and complete fermentation of the monosaccharides released upon the lactose hydrolysis step (lab- and pilot-scale) and prior to the final membrane concentration step. An further improvement involved optimizing a nanofiltration membrane for rejection/retention coefficients of sialyl-oligosaccharides/simple sugars at various pH and transmembrane pressure values. Carbohydrates were quantified nanofiltration, as well as nanofiltration combined with diafiltration were successful for isolation of using high performance HPAE-PAD, and Chip Q ToF was utilized to obtain a complete glycan profile. We discovered that pH and operating pressure during filtration have profound effects on the speed of filtration and selectivity membrane. Oligosaccharide purity was improved from 4% in the colostrum whey permeate to 98% in the final retentate following diafiltration, with 99.8% permeation of simple sugars and nearly 96% recovery of oligosaccharides. Over 2 kg of purified materials were produced using this combined strategy; respectively 1.1 kg of bovine milk oligosaccharides and 900 g of human milk oligosaccharides. Nanofiltration of hydrolyzed colostrum whey permeate with subsequent diafiltrations to remove monosaccharides is shown to be a reliable method for enrichment of bovine milk oligosaccharides with the potential application to conventional bovine whey permeate from whey protein isolation, as well to other mammalian milks.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Guerrero, A., L. Lerno, D. Barile, C.B. Lebrilla. Top-Down Analysis of highly post-translationally modified peptides by Electrospray Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry. J. Am. Soc. Mass Spectrom., 26(3): 453-459.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Dallas D., A. Guerrero, E. Parker, R. Robinson., J. Gan, J.B. German, D. Barile, C.B. Lebrilla. Current peptidomics: applications, purification, identification, quantification and functional analysis. Proteomics, 15(5-6): 1026-1038.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: D. Dallas, C. Smink, R. Robinson., T. Tian , A. Guerrero , E. Parker , J. Smilowitz, K. Hettinga, M. Underwood, C.B. Lebrilla, J.B. German, D. Barile. Endogenous human milk Peptide release is greater after preterm birth than term birth. American Society for Nutrition, 145(3): 425-433.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Karav S., A. Le Parc, J.M.L.N. de Moura Bell, C. Rouqui�, D. A. Mills, D. Barile, D.E. Block. Kinetic Characterization of a novel Endo-?-N-acetylglucosaminidase on concentrated bovine colostrum whey. Enzyme and microbial technology, 77: 46-53.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: H Lee, VL de MeloSilva, Y Liu, D. Barile. Quantification of carbohydrates in whey permeate products using high-performance anion-exchange chromatography with pulsed amperometric detection. Journal of Dairy Science, 98(11): 7644-7649.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: DC Dallas, F Citerne, T Tian, VLM Silva, KM Kalanetra, SA Frese, RC Robinson, DA Mills, D. Barile. Peptidomic analysis reveals proteolytic activity of kefir microorganisms on bovine milk proteins. Food Chemistry, 197: 273284.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: A Mart�n-Ortiz, J Salcedo, D. Barile, A Bunyatratchata, FJ Moreno, I Martin-Garc�a, A Clemente, ML Sanz, AI Ruiz-Matute. Characterization of goat colostrum oligosaccharides by nano-liquid chromatography on chip quadrupole time-of-flight mass spectrometry and hydrophilic interaction liquid chromatography-quadrupole mass spectrometry. Journal of Chromatography A, doi:10.1016/j.chroma.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: AL Parc, S Karav, JMLND Bell, SA Frese, Y Liu, DA Mills, DE Block, D. Barile. A novel endo???N?acetylglucosaminidase releases specific N?glycans depending on different reaction conditions. Biotechnology progress, 31(5): 1323-1330.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: CC Hsieh, B Hern�ndez-Ledesma, S Fern�ndez-Tom�, V Weinborn, D. Barile, JMLN de Moura Bell. Milk Proteins, Peptides, and Oligosaccharides: Effects against the 21st Century Disorders. BioMed research international, Article ID 146840.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: AW Chassy, C Bueschl, H Lee, L Lerno, A Oberholster, D. Barile, R Schuhmacher, AL Waterhouse. Tracing flavonoid degradation in grapes by MS filtering with stable isotopes. Food chemistry, 166: 448-455.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Dallas, D.C., Sanctuary, M.R., Qu, Y., Khajavi, S.H., Zandt, A.E.V., Dyandra, M., Frese, S.A., Barile, D., and German, J.B. Personalizing Protein Nourishment. Critical Reviews in Food Science and Nutrition.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Karav, S., Bell, J.M.L.N.D.M., Parc, A.L., Liu, Y., Mills, D.A., Block, D.E., and Barile, D. Characterizing the release of bioactive N-glycans from dairy products by a novel endo-?-N-acetylglucosaminidase. Biotechnol Progress, 31: 13311339.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2015 Citation: Charbonneau, M., O'Donnell, D., Blanton, LV., Totten, SM., Davis, J.C.C., Barratt, M.J., Cheng, J., Guruge, J., Talcott, M., Bain, J.R., Muehlbauer, M.J., Ilkayeva, O., Wu, C., Struckmeyer, T., Barile, D., Mangani, C., Jorgensen, J., Fan, Y.M., Maleta, K., Dewey, K.G., Ashorn, P., Newgard, C.P., Lebrilla, C.B., Mills, D.A., Gordon, JI. Sialylated milk glycans promote growth in gnotobiotic mice and pigs with a stunted Malawian infant gut microbiota. Cell.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: March 15, Seminar: Personalizing Foods, Nutrigenomics, "FOOD IS A CONVERSATION", Food Innovation Program, Reggio Emilia, IT.
  • Type: Book Chapters Status: Published Year Published: 2015 Citation: Cohen J.L., M. Geissler, D. Barile, C.W. Simmons, J.M.L.N. de Moura Bell: Current Utilization of Dairy Industry Co-products; in Camille N. Foster (editor in chief) Agricultural Wastes: Characteristics, Types and Management. 978-1-63482-376-0, Nova Publishers.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: December 1, Guest Lecture: Milk Bioactive compounds at the interface of food and health, Graduate Group In Nutritional Biology & Western Human Nutrtion Research Center, UC Davis.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Invited talk: New opportunities and challenges for bioactive ingredients manufacturing, One Health Center Graduate Education Conference, Western Institute for Food Safety and Security, UC Davis January 24, 2015.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: March 19, Invited talk: Food digestion: what milk can teach us from the infant's perspective, 4th International Conference on Food Digestion, Naples, IT
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: March 25, Invited seminar: Functional Properties and Sources of Recently Identified Bioactive Food Components, Future Food Institute, University of Modena and Reggio Emilia, IT.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: March 26, Key Note Lecture: Taking lessons from Nature: the "omics" of bioguided processing for personalized health, Food Innovation Program, Reggio Emilia, IT
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: March 26, Invited seminar: Personalizing Foods & Nutrigenomics, Future Food Institute, University of Modena and Reggio Emilia, IT
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: May 28, Invited seminar: Beyond essential nutrition: Cultivating friendly bacteria with old and new prebiotics, IMB Research Distinguished Speaker series, Accelerated Discovery Lab, IBM Almaden Research Center, San Jose CA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: April 14, Invited talk: Novel tools to evaluate bioactive compounds in dairy products, Nanjing Agricultural University-UC Davis One Health Food Safety Symposium.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: May 12, Invited talk: Integrating Biology and New 'Omics to guide Diet, Health and Agriculture"., FAPESP-UC Davis Research Symposium in S�o Paulo (BR).
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: May 21, New technologies to discover new bioactive compounds in foods, World Bank  Haiti  Coffee Production. Alumni Center, Allewelt Conference Room, UC Davis.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: July 2, Guiding the Food supply to consumers, Campus visit by Mengniu Dairy Group Co. Ltd UC Davis Conference Center.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: July 21, Invited talk: Processing strategies for the purification of oligosaccharides from donor milk and agricultural streams, FASEB Science Research Conferences: Origins and Benefits of Biologically-Active Components of Human Milk. Big Sky, MT, USA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: August 11, Lecture: A roadmap for milk bioactives RMI & Beijing University of Chemical Technology, China.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: October 26, Moderated industry panel on "Impacts of Microbiomes" at the FALL 2015 University-Industry Consortium Meeting  UC Davis Conference Center.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: December 10, invited talk: Beyond essential nutrition: discovering & delivering bioactive compounds, John E. Kinsella Memorial Symposium and Lecture The Fitzgerald Debating Chamber, University College Dublin (Ireland)
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Hallam, M., D. Barile, M. Meyrand, J.B. German, J.B., R. Reimer. Maternal high?protein or high?prebiotic?fiber diets affect maternal milk composition and gut microbiota in rat dams and their offspring. Obesity, 22(11): 2344-2351.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Holton, T., V. Vaishnavi, D. Dallas, A. Guerrero, R. Borghese, C. Lebrilla, J.B. German, D. Barile, M. Underwood, D. Shields, N. Khaldi. Following the digestion of milk from mother to baby. Journal of Proteome Research. Journal of proteome research, 13(12): 5777-5783.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Maldonado-Gomez, M., H. Lee, D. Barile, M. Lu., R. Goodman., D. Rose., R. Hutkins. Adherence inhibition of enteric pathogens to epithelial cells by bovine colostrum oligosaccharides and other bovine colostrum fractions. International Dairy Journal. International Dairy Journal, 40: 24-32.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Pacheco A., D. Barile , M. Underwood. A., D. Mills. The Impact of the Milk Glycobiome on the Neonate Gut Microbiota, Annual Review of Animal Science. Vol. 3. 3: 419-445.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Guerrero, A., D.C. Dallas, S. Contreras, A. Bhandari , A. Canovas, A. Islas-Trejo, J.F. Medrano , E.A. Parker, D. Barile, J.B. German. Peptidomic analysis of healthy and mastitic bovine milk. International Dairy Journal, 46: 46-52.


Progress 10/01/13 to 09/30/14

Outputs
Target Audience: Our target audience is the California food industry. Oligosaccharides, peptides and glycoproteins represent a new opportunity to prevent or ameliorate major infectious and chronic disease worldwide. Our research helps valorize agricultural by- products from the diary the produce industries. I am committed to translating and communicating scientific knowledge to fellow scientists and the general public. To date I have addressed the industry both in small and large meetings, and have presented my work at over 10 conferences attended by industry participants. During the reporting period I presented this research to the following companies: Merk, DSM, Roll Global, Pentair, DSM, Valio, Leprino cheese, IndoFood, Proliant, California Research Alliance, California Dairies, Constellation Brands, CreAgri, SomaLogic, Rizo Lopex, GenTech China, Nestle', Glambia, Abbott, Clorox, Don Francisco, El Mexicano and more. To enable translation of the research to food products, I also have active projects with Hilmar Ingredients, Danone, LaBelle Inc., Sterling Technology, Prolacta Bioscience, Tamarak Biotics, Davisco, Green Mountain Coffee, HiU Coffee, Vitalus Nutrition and Arla Foods. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? 10 undergraduates, 3 PhD student, and two visiting professors were trained on our methods. We have established relationships with large manufacturers of pilot-scale and industrial filtration equipment that are interested in taking our prototype to commercial reality, and we are seeking partnership with industries in order to pack and distribute the supplement in the functional foods market. I visited our collaborators (mostly large dairy companies based in California) and disseminated our research results. I also presented this research to 14 national and international conferences. During this review period, I served as Associate Editor for the Newsletter "SPLASH!" distributed by the International Milk Genomic Consortium and I contributed 12 pieces to educate the industry and the general public in milk research. I have published 3 additional papers in high-impact, editorially reviewed articles read by our stakeholders (food industry, food processors). I was interviewed about my research by: PBS, Discovery Magazine, The New York Times, Dairy Foods, CE&N, Science News ,Scientific American, and, the Washington Post. How have the results been disseminated to communities of interest? We have established relationships with large manufacturers of pilot-scale and industrial filtration equipment that are interested in taking our prototype to commercial reality, and we are seeking partnership with industries in order to pack and distribute the supplement in the functional foods market. I visited our collaborators (mostly large dairy companies based in California) and disseminated our research results. I also presented this research to 14 national and international conferences. During this review period, I served as Associate Editor for the Newsletter "SPLASH!" distributed by the International Milk Genomic Consortium and I contributed 12 pieces to educate the industry and the general public in milk research. I have published 3 additional papers in high-impact, editorially reviewed articles read by our stakeholders (food industry, food processors). I was interviewed about my research by: PBS, Discovery Magazine, The New York Times, Dairy Foods, CE&N, Science News ,Scientific American, and, the Washington Post. What do you plan to do during the next reporting period to accomplish the goals? We plan to develop a commercially feasible reactor to capture bioactive endogenous milk peptides and bioactive oligosaccharides currently wasted in dairy processing streams as bovine whey permeate. Our proposed reactor will be able to extract valuable components known to inhibit the activity of pathogenic bacteria and modulate the immune system, from industrial streams. This process will bring greater financial value to dairy waste, improving industry profits, while reducing environmental concerns with waste disposal.

Impacts
What was accomplished under these goals? During this reporting period my group published 28 peer-reviewed journal articles as well as 5 chapters in specialized books. Our work expanded from free oligosaccharides and peptided to include glycoproteins. We revealed a high diversity for N-glycan structures in goat milk lactoferrin that may help infants' health by decreasing pathogen infection. The analytical platform used enabled a comprehensive profiling of N-glycans, of a protein purified from a complex mixture, with high resolution and separation of isomeric forms. The N-glycan compositions were identified by an automated data analysis using a theoretical library for goat milk glycoproteins. This library was created combining biological rules and previous knowledge on milk N-glycome; the library will find applications in the N-glycan identification for other glycoproteins in goat milk or other sources. This study contributed to increase the knowledge on goat milk lactoferrin and showed that goat milk lactoferrin is a good candidate to supplement bovine based infant formula because of the homology in the N-glycan composition with its human counterpart. We already demonstrated that the techniques chosen for the isolation (our prototype) are well suited to deliver functional compounds from dairy waste streams. We showed that bovine milk contains endogenous peptides that are being tested for their ability to inhibit the growth of pathogens representative of the gram-negative and gram-positive bacterial classes. We successfully set up a pilot'scale method capable of processing over 300L of dairy streams/day and we generated, for the first time, 1 kg of pure oligosaccharides that have already demonstrated a really interesting behavior with regard to immunomodulation.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2012 Citation: Bordiga M., Travaglia F., Meyrand M German J.B., Lebrilla C.B.,Co�sson J.D.,Arlorio M., Barile D. (2012). Identification and Characterization of Complex Bioactive Oligosaccharides in White and Red Wine by a Combination of Mass Spectrometry and Gas Chromatography. J. Agric Food Chem 60: 3700-3707. PMC3334496
  • Type: Journal Articles Status: Accepted Year Published: 2012 Citation: Strum J.S., Aldredge D., Barile D., Lebrilla C.B. (2012). Coupling Flash LC with MS for enrichment and isolation of milk oligosaccharides for functional studies. Analytical Biochemistry 424:87-96. PMC3562133
  • Type: Journal Articles Status: Accepted Year Published: 2012 Citation: Garrido D., Barile D., Mills D.A. (2012). A molecular basis for bifidobacterial enrichment in the infant gastrointestinal tract. Advances in Nutrition; vol. 3: 415S-421S. PMC3649478
  • Type: Journal Articles Status: Accepted Year Published: 2012 Citation: Sundekilde U., Barile D., Meyrand M., Poulsen N., Larsen L., Lebrilla C.B., German J.B, Hanne B. (2012). Natural variability in bovine milk oligosaccharides from Danish Jersey and Holstein-Friesian breeds. Journal of Agricultural and Food Chemistry, 60(24):6188-96. PMC3386800
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Montella R., Co�sson J.D., Travaglia F., Locatelli M., Bordiga M., Meyrand M., Lebrilla C.B., Barile D., Arlorio M. (2013). Identification and Characterization of Water and Alkali Soluble Oligosaccharides from Hazelnut Skin (Corylus avellana L.). Food Chemistry. 140 (4), 717725.
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Aldredge D.L., Geronimo M.R, Hua S.S., Nwosu C.C., Lebrilla C.B. & Barile D. (2013). Annotation and structural elucidation of bovine milk oligosaccharides and determination of novel fucosylated structures. Glycobiology 23(6):664-76
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Dallas, D.; Guerrero, A.; Khaldi, N.; Castillo, P.; Martin, W.; Smilowitz, J.; Bevins, C.; Barile, D.; German, J.B; Lebrilla, C. (2013). Extensive in vivo human milk peptidomics reveals specific proteolysis yielding protective antimicrobial peptides. Journal of Proteome Research, 12 (5), 22952304.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: Barile D. Analytics for the discovery of bioactive compounds in agricultural products and by-products. Coffee Center Research Conference, UC Davis Conference Center, March 11, 2014
  • Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: Barile D. Prebiotic Oligosaccharides in human and bovine milks: isolation, characterization and functional testing. The 10th Vahouny Dietary Fiber Symposium, March 26-28, 2014 Washington DC.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2014 Citation: Barile D. Chemistry, Biology & Evolution to design new food ingredients. California State University, Chico, May 2nd 2014
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Smilowitz J., OSullivan A., Barile D., German J.B., L�nnerdal B., Slupsky C.M. (2013). The Human Milk Metabolome Reveals Diverse Oligosaccharide Profiles. The Journal of Nutrition. 143(11):1709-18
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Lee H.; German J.B.; Kjelden R.; Lebrilla C.B.; Barile D. (2013). Quantitative Analysis of Bovine Milk Gangliosides by Ultra-high Performance Liquid Chromatography-Tandem Mass Spectrometry. J. Agr. Food Chem. 61(40):9689-96.
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Underwood MA, Kalanetra KM, Bokulich NA, Mirmiran M, Barile D., Tancredi DJ, German JB, Lebrilla CB, Mills DA. (2013). Prebiotic Oligosaccharides in Premature Infants. In press at Journal of Pediatric Gastroenterology and Nutrition 2013 Oct 16.
  • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Le Parc A., Lee H., Chen K. and Barile D. (2014). Rapid quantification of functional carbohydrates in food products. Food and Nutrition Sciences, 5: 71-78.
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Dallas, D., Guerrero D., Parker E., Garay L., Bhandari A., Lebrilla C.B., Barile D., German, J.B. (2013). Peptidomic profile of milk of Holstein cows at peak lactation. Journal of Agricultural and Food Chemistry 62 (1), 5865.
  • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Le Parc A., Dallas D.C., Duaut S., Leonil J., Martin P., Barile D. (2014). Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. Electrophoresis 35 (11), 1560-1570.
  • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Lee H., Garrido D., Mills D.A., Barile D.* (2014). Hydrolysis of milk gangliosides by infant-gut associated bifidobacteria determined by microfluidic chips and high-resolution mass spectrometry. Electrophoresis 35 (11), 17421750.
  • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Dallas DC., Guerrero A., Khaldi N., Borghese R., Bhandari A., Underwood M.A., Lebrilla C.B., German J.B., Barile D. (2014). A peptidomic analysis of human milk digestion in the infant stomach reveals protein-specific degradation patterns The Journal of nutrition 144 (6), 815-820.
  • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Domizio P., Liu Y., Bisson L., Barile D.* (2014). Use of non-Saccharomyces wine yeast as novel sources of mannoproteins in wine. Food Microbiology 43 (2014) 5-15.
  • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Mehra R, Barile D.*, Marotta M., Lebrilla C. B., Chu C., German J. B. (2014). Novel High-Molecular Weight Fucosylated Milk Oligosaccharides Identified in Dairy Streams. PlosOne, 9 (5)
  • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Chassy, A., Bueschl, C., Lee, H., Lerno, L., Oberholster, A., Barile, D., Schuhmacher, R., Waterhouse, A. Tracing flavonoid degradation in grapes by MS filtering with stable isotopes. Food Chem. 166:448-55.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2014 Citation: Hallam, M., Barile, D., Meyrand, M., German, J.B., and Reimer, R. Maternal diets high in protein or prebiotic fiber differentially affect maternal milk composition and gut microbiota in rat dams and their offspring. Obesity (in press) doi:10.1002/oby.20849.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Pacheco A., D. Barile, M. Underwood. A. D. Mills. 2015. The Impact of the Milk Glycobiome on the Neonate Gut Microbiota, Annual Review of Animal Science, in press, DOI: 10.1146/annurev-animal-022114-111112
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Maldonado-Gomez, M., H. Lee, D. Barile, M. Lu., R. Goodman., D. Rose, R. Hutkins. Adherence inhibition of enteric pathogens to epithelial cells by bovine colostrum oligosaccharides and other bovine colostrum fractions. International Dairy Journal 40: 2432.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Holton, T.A., Vijaykumar, V., Dallas, D.C., Guerrero, A., Borghese, R.C., Lebrilla, C.B., German, J.B., Barile, D., Underwood, M.A., Shields, D.C., Khaldi, N., 2014. Following the digestion of milk proteins from mother to baby. J. Proteome Res. 13 (12), pp 57775783
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Dallas D., A. Guerrero, E. Parker, R. Robinson., J. Gan, J.B. German, D. Barile, C.B. Lebrilla (2014) Current peptidomics: applications, purification, identification, quantification and functional analysis (2015) Journal of Proteomics. 2014 Nov 27. doi: 10.1002/pmic.201400310
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: D. Dallas, C. Smink, R. Robinson., T. Tian , A. Guerrero , E. Parker , J. Smilowitz, K. Hettinga, M. Underwood, C.B. Lebrilla, J.B. German, D. Barile (2015). Preterm birth increases endogenous human milk peptide release compared to term birth. Journal of Nutrition. In press.
  • Type: Book Chapters Status: Published Year Published: 2012 Citation: Nijman R.M., Meyrand M. & Barile D. (2012). Human milk oligosaccharides and their health effects. In: Lactation, Natural Processed, Physiological responses and role in Maternity. L. M. Reyes Cruz and D. C. Ortiz Gutierrez Editors. Nova Science Publishers Inc, New York. ISBN: 978-1-62257-6 pp. 43-71.
  • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Dallas D.C, Weinborn V., de Moura Bell JMLN, Wang M., Parker E.A., Guerrero A., Hettinga K.A, Lebrilla C.B, German J.B., Barile D.* (2014). Comprehensive peptidomic and glycomic evaluation reveals sweet whey permeate from colostrum is a source of milk protein-derived peptides and oligosaccharides. In press at Food Research International DOI: 10.1016/j.foodres.2014.03.021
  • Type: Book Chapters Status: Published Year Published: 2014 Citation: Dallas D.C., Meyrand M., Barile D. (2014). Production and bioactivity of bovine (and related species) milk oligosaccharides. In press at: Food Oligosaccharides' edited by F.J. Moreno & M.L. Sanz  IFT Press; Wiley-Blackwell Oxford ISBN: 978-1-118-42649-4
  • Type: Book Chapters Status: Published Year Published: 2014 Citation: Dallas D.C., Meyrand M., Barile D.* (2014). Production and bioactivity of bovine (and related species) milk oligosaccharides. In press at: Food Oligosaccharides' edited by F.J. Moreno & M.L. Sanz  IFT Press; Wiley-Blackwell Oxford ISBN: 978-1-118-42649-4 pp.21-34
  • Type: Book Chapters Status: Published Year Published: 2014 Citation: Lange M, Dallas DC, Lee HY, Le Parc A, Nobrega JM, Barile D.* Human Nutrition: Determining functional properties and sources of recently identified food components: oligosaccharides, glycolipids, glycoproteins and peptides. (2014). In: Neal Van Alfen, editor-in-chief. Encyclopedia of Agriculture and Food Systems, Vol. 2, San Diego: Elsevier; 2014. pp. 441-461.
  • Type: Book Chapters Status: Awaiting Publication Year Published: 2015 Citation: C.C. Hsieh, B. Hern�ndez-Ledesma, S. Fern�ndez-Tom�, V. Weinborn, D. Barile and J.M.N. Nobrega de Moura Bell. (2015). Milk Proteins, Peptides And Oligosaccharides: Effects Against The 21st Century Disorders. In: Food Bioactive Compounds against Diseases of the 21st Century, Hindawi Publishing Corporation
  • Type: Conference Papers and Presentations Status: Other Year Published: 2012 Citation: Barile D. Complex oligosaccharides as dairy ingredients Dairy Ingredients Symposium, San Francisco March 1-3, 2012
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Omics, and maggots, and mites ..oh my. UCD/Agilent Joint Halloween Symposium: Return of the mummy: New technologies for measuring very old things, Buehler Alumni Center October 31, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2012 Citation: Barile D. FOODS FOR HEALTH:The Milk Bioactive Program. UC Davis Licensing Academy, June 6-2012
  • Type: Conference Papers and Presentations Status: Other Year Published: 2012 Citation: Barile D. Bioguided food processing of agricultural by- products for personalized health, IFT 13 Annual meeting Las Vegas June 23-28. 2012
  • Type: Conference Papers and Presentations Status: Other Year Published: 2103 Citation: Barile D. Mining bioactive molecules from food processing sidestreams: Promoting health and environmental sustainability. Joint Symposium of UC Davis and Academia Sinica Taiwan on Chemical Biology December 2 and 3, 2013 Activities and Recreation Center, Ballroom A, UC Davis
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Effects of consuming milk oligosaccharides (extracted from cheese whey) on gut microbiota and tolerability in healthy humans. International Milk Genomics Consortium Symposium, UC Davis Conference Center. October -3, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: DeMoura J. & Barile D. Recovering Valuable Components from Dairy By-products. "Managing Energy, Water and By-products for Profitability in Food and Beverage Processing. CIFAR annual conference, Silverado Sensory Theater UC Davis. October 10, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Discovery of bioactive molecules in industrial side-streams to Improve Health" Calpoly Feb 28, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2012 Citation: Barile D. Novel prebiotics: glycans from dairy streams. CIFAR annual meeting, UC Davis, April 18, 2012
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Bioguided Food Processing: Harvesting Ingredients from Side-Streams to Improve Health. Davis Rotary Club, February 15, 2013.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Cesena, Comprehensive evaluation of foods bioactive components using a combination of Proteomics, Glycomics, Lipidomics and Peptidomics. Foodomics, the science for discovering, Cesena, Italy, May 22-24, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. combination of engeneering and analytics to discover bioactive food molecules. Universty of Oriental Piedmot, Novara, Italy, graduate students seminar series, May 28 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Identification and characterization of prebiotics in cheese whey. Deans Advisory Council, UC Davis, April 18, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Effects of consuming milk oligosaccharides (extracted from cheese whey) on gut microbiota and tolerability in healthy humans. International Milk Genomic Symposium, Davis October 1-3, 2013


Progress 10/01/12 to 09/30/13

Outputs
Target Audience: Our target audience is the California food industry. Oligosaccharides and peptides represent a new opportunity to prevent or ameliorate major infectious and chronic disease worldwide. Our research helps valorize agricultural by- products from the diary the produce industries. During the reporting period I presented this research to the following companies: Hilmar Ingredients, LaBelle Inc., Sterling Technology, Prolacta Bioscience, El Mexicano, POM Wonderful, Don Francisco, Leprino cheese, INDOFood, Proliant, TAMARAK BIOTICS, Davisco, Constellation wines, CreAgri Olive oil. The goal of this project is identifying bioactive oligosaccharides, glycoproteins and other bioactive compound in sources other than human milk. Much progress was made during this reporting period including an unexpected and much desirable discovery. The proposed work included a first step of enrichment by pilot scale membrane filtration (combination of ultrafiltration and nanofiltration) of dairy streams followed by more specific separations and analysis by High-Accuracy Mass Spectrometry combined with Nanoliquid Chromatography. Whereas our experiments with membrane filtration and chromatography did not detect any glycoproteins or glycolipids in dairy streams, we found and characterized a myriad of oligosaccharides and bioactive peptides. In particular, our discovery that bioactive peptides are present in whey permeate (a dairy waste product) has opened up a great opportunity to extract them from agricultural streams in large amounts by coupling the concept of molecular weight fractionation and large-scale solid phase extraction. Capturing bioactive peptides and oligosaccharides from the massive amounts of dairy waste streams produced in California represents an unprecedented opportunity to develop the next generation of microbiome and immune system modulating therapeutics while solving economic and environmental problems associated with disposal and treatment of industrial streams. As an example, the California dairy industry produces over 50 million pounds of waste streams daily. We have identified a specific process that has the potential to capture valuable components present in these dairy waste streams. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? 7 undergraduates, 1 PhD student, and one visiting professor were trained on our methods. How have the results been disseminated to communities of interest? We have established relationships with large manufacturers of pilot-scale and industrial filtration equipment that are interested in taking our prototype to commercial reality, and we are seeking partnership with industries in order to pack and distribute the supplement in the functional foods market. I visited our collaborators (mostly large dairy companies based in California) and disseminated our research results. I also presented this research to 14 national and international conferences. What do you plan to do during the next reporting period to accomplish the goals? We plan to develop a commercially feasible reactor to capture bioactive endogenous milk peptides and bioactive oligosaccharides currently wasted in dairy processing streams as bovine whey permeate. Our proposed reactor will be able to extract valuable components known to inhibit the activity of pathogenic bacteria and modulate the immune system, from industrial streams. This process will bring greater financial value to dairy waste, improving industry profits, while reducing environmental concerns with waste disposal.

Impacts
What was accomplished under these goals? During the past year our work at the Milk Processing Lab was focused on the optimization of peptides and carbohydrates quantification and characterization to obtain purified fractions that will be used in multiple assays for functional testing. We already demonstrated that the techniques chosen for the isolation (our prototype) are well suited to deliver functional compounds from dairy waste streams. We showed that bovine milk contains endogenous peptides that inhibit the growth of pathogens representative of the gram-negative and gram-positive bacterial classes.

Publications

  • Type: Journal Articles Status: Published Year Published: 2012 Citation: Bordiga M., Travaglia F., Meyrand M German J.B., Lebrilla C.B.,Co�sson J.D.,Arlorio M., Barile D. (2012). Identification and Characterization of Complex Bioactive Oligosaccharides in White and Red Wine by a Combination of Mass Spectrometry and Gas Chromatography. J. Agric Food Chem 60: 3700-3707. PMC3334496
  • Type: Journal Articles Status: Published Year Published: 2012 Citation: Strum J.S., Aldredge D., Barile D., Lebrilla C.B. (2012). Coupling Flash LC with MS for enrichment and isolation of milk oligosaccharides for functional studies. Analytical Biochemistry 424:87-96. PMC3562133
  • Type: Journal Articles Status: Published Year Published: 2012 Citation: Garrido D., Barile D., Mills D.A. (2012). A molecular basis for bifidobacterial enrichment in the infant gastrointestinal tract. Advances in Nutrition; vol. 3: 415S-421S. PMC3649478
  • Type: Journal Articles Status: Published Year Published: 2012 Citation: Sundekilde U., Barile D., Meyrand M., Poulsen N., Larsen L., Lebrilla C.B., German J.B, Hanne B. (2012). Natural variability in bovine milk oligosaccharides from Danish Jersey and Holstein-Friesian breeds. Journal of Agricultural and Food Chemistry, 60(24):6188-96. PMC3386800
  • Type: Journal Articles Status: Published Year Published: 2012 Citation: Barile D. (2012). New microbiota pathways. The world of Food Ingredients, 98-101.
  • Type: Journal Articles Status: Awaiting Publication Year Published: 2014 Citation: Le Parc A., Dallas D.C., Duaut S., Leonil J., Martin P., Barile D. (2014). Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. In press at ELECTROPHORESIS (elps.201300619)
  • Type: Book Chapters Status: Published Year Published: 2012 Citation: Nijman R.M., Meyrand M. & Barile D. (2012). Human milk oligosaccharides and their health effects. In: Lactation, Natural Processed, Physiological responses and role in Maternity. L. M. Reyes Cruz and D. C. Ortiz Gutierrez Editors. Nova Science Publishers Inc, New York. ISBN: 978-1-62257-6 pp. 43-71.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Omics, and maggots, and mites ..oh my. UCD/Agilent Joint Halloween Symposium: Return of the mummy: New technologies for measuring very old things, Buehler Alumni Center October 31, 2013
  • Type: Journal Articles Status: Other Year Published: 2013 Citation: Barile D. Effects of consuming milk oligosaccharides (extracted from cheese whey) on gut microbiota and tolerability in healthy humans. International Milk Genomics Consortium Symposium, UC Davis Conference Center. October -3, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: DeMoura J. & Barile D. Recovering Valuable Components from Dairy By-products. "Managing Energy, Water and By-products for Profitability in Food and Beverage Processing. CIFAR annual conference, Silverado Sensory Theater UC Davis. October 10, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Discovery of bioactive molecules in industrial side-streams to Improve Health" Calpoly Feb 28, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2012 Citation: Barile D. Novel prebiotics: glycans from dairy streams. CIFAR annual meeting, UC Davis, April 18, 2012
  • Type: Book Chapters Status: Awaiting Publication Year Published: 2014 Citation: 2. Dallas D.C., Meyrand M., Barile D. (2014). Production and bioactivity of bovine (and related species) milk oligosaccharides. In press at: Food Oligosaccharides' edited by F.J. Moreno & M.L. Sanz  IFT Press; Wiley-Blackwell Oxford ISBN: 978-1-118-42649-4 (In press, expected publication: May 2014).
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Mining bioactive molecules from food processing sidestreams: Promoting health and environmental sustainability. Joint Symposium of UC Davis and Academia Sinica Taiwan on Chemical Biology December 2 and 3, 2013 Activities and Recreation Center, Ballroom A, UC Davis
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Bioguided Food Processing: Harvesting Ingredients from Side-Streams to Improve Health. Davis Rotary Club, February 15, 2013.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Cesena, Comprehensive evaluation of foods bioactive components using a combination of Proteomics, Glycomics, Lipidomics and Peptidomics. Foodomics, the science for discovering, Cesena, Italy, May 22-24, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: 9. Barile D. combination of engineering and analytics to discover bioactive food molecules. Universty of Oriental Piedmot, Novara, Italy, graduate students seminar series, May 28 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Identification and characterization of prebiotics in cheese whey. Deans Advisory Council, UC Davis, April 18, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2013 Citation: Barile D. Effects of consuming milk oligosaccharides (extracted from cheese whey) on gut microbiota and tolerability in healthy humans. International Milk Genomic Symposium, Davis October 1-3, 2013
  • Type: Conference Papers and Presentations Status: Other Year Published: 2012 Citation: Barile D. Complex oligosaccharides as dairy ingredients Dairy Ingredients Symposium, San Francisco March 1-3, 2012
  • Type: Conference Papers and Presentations Status: Other Year Published: 2012 Citation: 13. Barile D. FOODS FOR HEALTH:The Milk Bioactive Program. UC Davis Licensing Academy, June 6-2012
  • Type: Conference Papers and Presentations Status: Other Year Published: 2012 Citation: Barile D. Bioguided Food Processing for Personalized Health. IFT annual meeting, Las Vegas, June 26, 2012