Source: UNIVERSITY OF CALIFORNIA, DAVIS submitted to NRP
SYNERGY BETWEEN MILK AND PROBIOTICS FOR GASTROINTESTINAL HEALTH
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1011897
Grant No.
2017-67017-26532
Cumulative Award Amt.
$469,257.00
Proposal No.
2016-08943
Multistate No.
(N/A)
Project Start Date
May 1, 2017
Project End Date
Jun 30, 2022
Grant Year
2017
Program Code
[A1341]- Food Safety, Nutrition, and Health: Function and Efficacy of Nutrients
Recipient Organization
UNIVERSITY OF CALIFORNIA, DAVIS
410 MRAK HALL
DAVIS,CA 95616-8671
Performing Department
Food Science & Technology
Non Technical Summary
Although dairy is an important pillar of a healthy diet, the majority of the U.S. population has a low intake of dairy along with other key food groups including vegetables, fruits, and whole grains. One opportunity for increasing dairy consumption is through fermented dairy products. Even though fluid milk constitutes the largest fraction of dairy consumed by the US population, intake of fermented dairy products such as yogurt and cheese is increasing. This change in consumer behavior presents an opportunity for augmentation of the inherent benefits of milk through the incorporation of probiotic bacteria. Numerous studies performed in vitro and in vivo indicate that fermented dairy foods, and probiotic-containing dairy products in particular, augment the benefits of dairy in promoting human health. Therefore, we will apply mechanistic studies to elucidate the dairy-probiotic connection in human health. Our efforts will include studies designed to measure how dairy foods can be applied to maintain healthy intestinal function, support glucose homeostasis, and confer normal (as opposed to anxiety-induced) cognition/behaviors, even under times of stress. The results will be evaluated and disseminated through peer-reviewed journal articles as well as at scientific conferences and meetings with the public and consumer interest groups.
Animal Health Component
(N/A)
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
70234501010100%
Knowledge Area
702 - Requirements and Function of Nutrients and Other Food Components;

Subject Of Investigation
3450 - Milk;

Field Of Science
1010 - Nutrition and metabolism;
Goals / Objectives
The over-arching goal of this project is to understand the mechanisms by which probiotic bacteria and the dairy matrix work synergistically to promote gastrointestinal and systemic health. The goal will be achieved by the following specific objectives: 1. Determine the capacity of L. casei and milk to prevent intestinal inflammation, accelerate recovery time, and prevent cognitive/behavioral deficits after exposure to an intestinal irritation in lean mice; 2. Determine the capacity of L. casei and milk to prevent intestinal and systemic inflammation, accelerate recovery time, and prevent cognitive/behavioral deficits after exposure to an intestinal irritant in obese mice; 3. Quantify the anti-inflammatory and barrier function effects of milk-grown L. casei on human cells
Project Methods
Two of the three objectives encompass rodent studies to determine the underlying mechanistic effects of dairy-probiotic combinations in people. Mice will be acclimated to a high fat, high sugar diet for either a short period of time (one week) or longer (10 weeks, including oral glucose tolerance test). The mice will then be grouped according to whether they are fed Lactobacillus casei in milk or nutrient-free buffer or milk / buffer alone. Several days after initiating the consumption of those dietary amendments, the mice will be exposed to an intestinal irritant for several more days. Mice will then be measured for behavioral changes including cognitive function and anxiety-like behavior. At the end of the study, the mice will be measured for colonic inflammation by histology, and a disease activity index as well as for intestinal permeability, immune responses, and intestinal microbiota. In the final objective, a mechanistic approach to understanding how L. casei and milk combinations alter health, we will also measure human epithelial cell line responses to these dietary additions. Epithelial barrier function and protein changes in response to perturbation will be measured with and without the milk and exogenous bacteria. This work is guided by the two hypotheses: (i) milk and L. casei function synergistically to reduce intestinal inflammation and associated cognitive and behavioral deficits that result from inflammatory processes; (ii) incubation of L. casei in milk improves the capacity of this probiotic to maintain the epithelial barrier integrity of human cells upon exposure to pro-inflammatory cytokines.

Progress 05/01/17 to 06/30/22

Outputs
Target Audience:The target audiences include individuals, groups, market segments, and communities that are interested in promoting human health through dietary approaches. This work is needed because milk and fermented dairy products are highly consumed foods that provide a high density of required nutrients. Incorporation of living probiotic bacteria in those foods can result in a cost-effective way to provide beneficial microbes to the consumer at reduced cost to those who are economically disadvantaged. Changes/Problems:We experienced delays to the research due to the COVID-19 pandemic. The pandemic caused significant hardship due to lab closures (March 2020 - July 2021) and supply/consumable delays (January 2021-June 2022). What opportunities for training and professional development has the project provided?This project has provided training to a postdoctoral fellow, a graduate student, undergraduate students, and technicians. Besides training on scientific methods and scientific writing, this project has also resulted in professional development opportunities including a presentation at scientific conferences. How have the results been disseminated to communities of interest?Dissemination events include conference presentations and consultations with local and international food processors and members of the media. Dissemination of knowledge beyond a scientific audience has included on-site tours of the research laboratory and description of the research program to university students, international visitors, casual visitors to the university (members of the public) and stakeholders (representing industrial sectors related to food science, nutrition, and health). What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? We showed that milk and probiotic L.casei - containing milk mitigate intestinal inflammation and sustain intestinal epithelial barrier integrity. The findings suggest that these beneficial effects are the result of interactions between the compounds in milk and L. casei - fermented milk with the intestinal epithelium. We also observed through the use of a pre-clinical mouse model that there are sex-dependent differences in intestinal responses which may be related to the enrichment of certain bacterial species in the gut microbiomes of female mice. The female mice had a much higher level of (DSS) colitis than the males and L. casei fermented milk conferred greater benefits to male mice. Additionally, we found that a fermentate (cell free L. casei fermented milk) increased intestinal barrier integrity during exposure to the INF-gamma cytokine. Examination of the human intestinal epithelial transcriptome in response to milk and the L. casei fermentate showed that the responses were specific to L. casei fermented milk and not other matrices. The fermentate reduced the expression of genes responsible for inflammatory responses to the cytokine. The fermentate also increased the expression of genes needed for sterol biosynthesis. These findings are important for providing accurate dietary guidance on the consumption of probiotic and dairy foods and for improving the health of US populations.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Invited Speaker. American Dairy Science Association Annual Meeting. 8-22-2021 Title: Dairy and probiotics pairing for intestinal health (via video)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Invited speaker. American Society for Nutrition Annual Conference. 6-10-2021. Fermented foods: more than a health food trend (via video)
  • Type: Conference Papers and Presentations Status: Published Year Published: 2021 Citation: Invited Speaker. Blue Hill and Stone Farms. 7-14-2021. Title: Eat your microbes! fermented foods, probiotics & digestive health (via video)
  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Maria L Marco, Robert Hutkins, Colin Hill, Victor L Fulgoni, III, Christopher J Cifelli, Jaime Gahche, Joanne L Slavin, Daniel Merenstein, Daniel J Tancredi, and Mary E Sanders A classification system for defining and estimating dietary intake of live microbes in US adults and children. Journal of Nutrition. 00:1-8
  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Robyn Campbell, Aviaja Hauptmann, Kristina Campbell, Shari Fox, Maria L. Marco. 2022. Better understanding of food and human microbiomes through collaborative research on Inuit fermented foods. Microbiome Research Reports. DOI: 10.20517/mrr.2021.06


Progress 05/01/20 to 04/30/21

Outputs
Target Audience:The target audiencesinclude individuals, groups, market segments, and communities that are interested in promoting human health through dietary approaches. This work is needed because milk and fermented dairy products are highly consumed foods that provide a high density of required nutrients. Incorporation of living probiotic bacteria in those foods can result in a cost-effective way to provide beneficial microbes to the consumer at reduced cost to those who are economically disadvantaged. Changes/Problems:We are experiencing delays to the research due to the COVID-19 pandemic. We are focused on data analysis and writing manuscripts, however, the pandemic has caused significant hardship since our research lab closed and has limited in-person access since March 2020. What opportunities for training and professional development has the project provided?This project has provided training to a postdoctoral fellow, a graduate student, undergraduate students, and technicians. Besides training on scientific methods and scientific writing, this project has also resulted in professional development opportunities including a presentation at scientific conferences. How have the results been disseminated to communities of interest?Dissemination events include conference presentations and consultations with local and international food processors and members of the media. Dissemination of knowledge beyond a scientific audience has included on-site tours of the research laboratory and description of the research program to university students, international visitors, casual visitors to the university (members of the public) and stakeholders (representing industrial sectors related to food science, nutrition, and health). What do you plan to do during the next reporting period to accomplish the goals?Our work will finalize efforts on elucidating the precise effects of dairy and probiotic combinations to mitigate intestinal inflammation. Firstly. we are currently completing the secondary analyses on the mouse studies. This work encompasses completing microbiota diversity analyses of the intestinal microbiota. We are also completing immune response and intestinal gene expression analyses from our murine model studies. Secondly, we will complete the cell culture analyses which have shown how milk-probiotic combinations increase intestinal barrier function when the epithelium is exposed to the IFN-gamma cytokine. Specifically, we will verify responses by focusing on quantification of levels of individual epithelial cell proteins identified to be responsive probiotic and milk combinations by transcript analysis. Manuscripts for two publications are underway and will be submitted for review and publication. These results will also be disseminated at (inter)national meetings, in small group meetings with industrial stakeholders, to the public by the press and other media outlets, and to academic groups by publishing peer-reviewed journal articles.

Impacts
What was accomplished under these goals? Accomplishments this year include the examination of milk and probiotic L. casei containing milk for the capacity to mitigate intestinal inflammation, sustain intestinal epithelial barrier integrity, and modulate behavior. We focused on identifying the gut microbiome response to milk, probiotic L. casei, and the combination of the two in a murine model of intestinal inflammation (colitis). We also completed our analysis of the mouse behavioral data and analysis of the mouse tissues by histology and ex vivo intestinal barrier function. Results of this work showed that the mice exhibited sex-specific responses. The female mice had a much higher level of colitis than the males and there were also differences in responses to milk - probiotic combinations between the sexes. Over the past year, we have also made progress with the analysis of intestinal epithelial responses to milk - probiotic combinations using a Caco-2 cell culture model. We found that a fermentate (cell free fermented milk) increased intestinal barrier function when Caco-2 cells were exposed to the INF-gamma cytokine. We are pursuing these findings because they show a clear and defined benefit of milk-probiotic combinations beyond the benefits of consuming milk alone.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Invited speaker, University of California, Riverside. Microbes in our foods: opportunities for improving human health. November 4, 2020.
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Maria L. Marco. 2021. Defining how microorganisms benefit human health. Microbial Biotechnology. 14 (1):35 -40 https://doi.org/10.1111/1751-7915.13685
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: . Maria L Marco, Colin Hill, Robert Hutkins, Joanne Slavin, Daniel J. Tancredi, Daniel Merenstein, and Mary Ellen Sanders. 2020 Should there be a recommended daily intake of microbes? The Journal of Nutrition. 150 (12): 3061 -3067 https://doi.org/10.1093/jn/nxaa323
  • Type: Journal Articles Status: Published Year Published: 2021 Citation: Maria L Marco, Mary Ellen Sanders, Michael G�nzle, Marie Claire Arrieta, Paul D. Cotter, Luc De Vuyst, Colin Hill, Wilhelm Holzapfel, Sarah Lebeer, Dan Merenstein, Gregor Reid, Benjamin E. Wolfe, and Robert Hutkins. 2021. Expert consensus document: The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on fermented foods. Nature Reviews Gastroenterology and Hepatology. https://doi.org/10.1038/s41575-020-00390-5


Progress 05/01/19 to 04/30/20

Outputs
Target Audience:The target audiencesinclude individuals, groups, market segments, and communities that are interested in promoting human health through dietary approaches. This work is needed because milk and fermented dairy products are highly consumed foods that provide a high density of required nutrients. Incorporation of living probiotic bacteria in those foods can result in a cost-effective way to provide beneficial microbes to the consumer at reduced cost to those who are economically disadvantaged. Changes/Problems:We are experiencing delays to the research due to the COVID-19 pandemic. We are focused on data analysis and writing manuscripts, however, the pandemic has caused significant hardship since our research lab closed in March. What opportunities for training and professional development has the project provided?This project has provided training to a postdoctoral fellow, a graduate student, undergraduate students, and technicians. Besides training on scientific methods and scientific writing, this project has also resulted in professional development opportunities including a presentation at scientific conferences. How have the results been disseminated to communities of interest?Dissemination events include conference presentations and consultations with local and international food processors and members of the media. Dissemination of knowledge beyond a scientific audience has included on-site tours of the research laboratory and description of the research program to university students, international visitors, casual visitors to the university (members of the public) and stakeholders (representing industrial sectors related to food science, nutrition, and health). What do you plan to do during the next reporting period to accomplish the goals?Our work will finalize efforts on elucidating the precise effects of dairy and probiotic combinations to mitigate intestinal inflammation. We are currently completing the secondary analyses on the mouse studies. and will prepare a manuscript on the results. The findings are informing our more detailed mechanistic studies on dairy-probiotic-gut interactions. These results will be disseminated at (inter)national meetings, in small group meetings with industrial stakeholders, to the public by the press and other media outlets, and to academic groups by publishing peer-reviewed journal articles.

Impacts
What was accomplished under these goals? Accomplishments this year include the examination of milk and probiotic L. casei containing milk for the capacity to mitigate intestinal inflammation, sustain intestinal epithelial barrier integrity, and modulate behavior. We used pre-clinical models and in vitro assays for these investigations with the goal of providing a mechanistic understanding of dairy food - probiotic interactions that benefit human health. Our findings support the hypothesis that probiotic efficacy depends on food delivery format. Our in vitro assessments indicate that dairy and probiotics work synergistically to benefit health. Our in vivo experiments have shown sex-specific and diet-dependent responses to milk and probiotic Lactobacillus casei. We have one manuscript underway and another we expect to complete by 2021.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Invited speaker. June 2019. Promoting healthy diets with fermented foods and beverages, Jiangnan University, Wuxi, China.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Invited Speaker. October 2019. Using Lactobacillus to understand and improve our intestinal ecosystems, Institute of genome sciences, University of Maryland, MD.
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Riley L. Hughes, Maria L. Marco, James P. Hughes, Nancy L. Keim, Mary E. Kable. 2019. The role of the gut microbiome in predicting response to diet and the development of precision nutrition models. Part I: Overview of Current Methods, Advances in Nutrition. https://doi.org/10.1093/advances/nmz022
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Katti Horng, Clarissa Santos-Rocha, Irina Grishina, Lauren Hirao, Anne Fenton, Sandipan Datta, Guochun Jiang, Maria L. Marco, Sumathi Sankaran-Walters, Cecilia Giulivi, Gino Cortopass, Satya Dandekar. 2019. Rapid reversal of gut epithelial barrier disruption in chronic SIV infection by L. plantarum through PPAR-mediated recovery of mitochondrial bioenergetics. Proceedings of the National Academy of Sciences. 116�(49)�24819-24829;�DOI:�10.1073/pnas.1908977116
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Invited Speaker. November 2019 Dietary modulation of the gut microbiota to improve and sustain health: probiotics. Nutrient delivery and impact on human health workshop sponsored by the International Center for Mechanical Sciences, Udine Italy
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Invited speaker. July 2019. Fermented foods and beverages. COSMOS course for high school students, Davis, CA.


Progress 05/01/18 to 04/30/19

Outputs
Target Audience:The target audiencesinclude individuals, groups, market segments, and communities that are interested in promoting human health through dietary approaches. This work is needed because milk and fermented dairy products are highly consumed foods that provide a high density of required nutrients. Incorporation of living probiotic bacteria in those foods can result in a cost-effective way to provide beneficial microbes to the consumer at reduced cost to those who are economically disadvantaged. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided training to a postdoctoral fellow, graduate student, undergraduate student, and technician. Besides training on scientific methods and scientific writing, this project has also resulted in professional development opportunities including a presentation at a scientific conference. How have the results been disseminated to communities of interest?Dissemination events include conference presentations and consultations with local and international food processors and members of the media. Dissemination of knowledge beyond a scientific audience has included on-site tours of the research laboratory and description of the research program to university students, international visitors, casual visitors to the university (members of the public) and stakeholders (representing industrial sectors related to food science, nutrition, and health). What do you plan to do during the next reporting period to accomplish the goals?Our work will focus on elucidating the precise effects of dairy and probiotic combinations to mitigate intestinal inflammation. We are currently completing the secondary analyses on the mouse studies. and will prepare a manuscript on the results. The findings are informing our more detailed mechanistic studies on dairy-probiotic-gut interactions. These results will be disseminated at (inter)national meetings, in small group meetings with industrial stakeholders, to the public by the press and other media outlets, and to academic groups by publishing peer-reviewed journal articles.

Impacts
What was accomplished under these goals? Accomplishments this year include the examination of milk and probiotic L. casei containing milk for the capacity to mitigate intestinal inflammation, sustain intestinal epithelial barrier integrity, and modulate behavior. We are using pre-clinical models and in vitro assays for these investigations with the goal of providing a mechanistic understanding of dairy food - probiotic interactions that benefit human health. Our findings support the hypothesis that probiotic efficacy depends on food delivery format. Our in vitro assessments indicate that dairy and probiotics work synergistically to benefit health. Our in vivo experiments have shown sex-specific and diet-dependent responses to milk and probiotic Lactobacillus casei

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Invited speaker. Title: Milk microbes: from the farm to human health. Department of Animal Science, University of CA, Davis. March 2019
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Panelist: Microbes associated with milk. International Milk Genomics Consortium Annual Meeting, Sacramento CA November 2018
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Zhengyao Xue, Mary E Kable, and Maria L Marco. 2018. Impact of DNA sequencing and analysis methods on 16S rRNA gene bacterial community analysis of dairy products. mSphere e00410-18.
  • Type: Journal Articles Status: Published Year Published: 2018 Citation: Sarah Lebeer, Peter A. Bron, Maria L. Marco, Jan-Peter Van Pijkeren, Mary O'Connell Motherway, Colin Hill, Bruno Pot, Stefan Roos, and Todd Klaenhammer. 2017. Identification of probiotic effector molecules: present state and future perspectives. Current Opinion in Biotechnology. 49:140 -147 https://doi.org/10.1016/j.copbio.2017.10.007
  • Type: Journal Articles Status: Published Year Published: 2019 Citation: Zhengyuan Zhai, Cristina Torres-Fuentes, Dustin D. Heeney, and Maria L Marco. 2019. Synergy between probiotic Lactobacillus casei and milk to maintain barrier integrity of intestinal epithelial cells. Journal of Agricultural and Food Chemistry. Jan 10. doi: 10.1021/acs.jafc.8b06657
  • Type: Conference Papers and Presentations Status: Published Year Published: 2019 Citation: Poster presentation. "Synergy between probiotic Lactobacillus casei and milk to maintain epithelial barrier integrity." Presented at the Northern California American Society for Microbiology Annual Meeting.


Progress 05/01/17 to 04/30/18

Outputs
Target Audience:The target audiencesinclude individuals, groups, market segments, or communities that are interested in promoting human health through dietary approaches. This is needed because milk and fermented dairy products are highly consumed foods that provide a high density of required nutrients. Incorporation of living probiotic bacteria in those foods can result in a cost-effective way to provide probiotics to the consumer at reduced cost to those who are economically disadvantaged. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided training to a postdoctoral fellow, graduate student, undergraduate student and technician. Besides training on scientific methods and scientific writing, this project has also resulted in professional development opportunities including a presentation at a scientific conference. How have the results been disseminated to communities of interest?Dissemination events include conference presentations and consultations with local and international food processors and members of the media. Dissemination of knowledge beyond a scientific audience has included on-site tours of the research laboratory and description of the research program to university students, international visitors, casual visitors to the university (members of the public) and stakeholders (representing industrial sectors related to food science, nutrition, and health). What do you plan to do during the next reporting period to accomplish the goals?Our work will continue to focus on elucidating the precise effects of dairy and probiotic combinations to mitigate intestinal inflammation. With first study nearly complete, we will be able to report on the findings and use those results to inform detailed mechanistic studies on dairy-probiotic-gut interactions. These results will be disseminated at (inter)national meetings, in small group meetings with industrial stakeholders, to the public by the press and other media outlets, and to academic groups by publishing peer-reviewed journal articles.

Impacts
What was accomplished under these goals? Accomplishments this year have included examination of milk and probiotic L. casei containing milk for the capacity to mitigate intestinal inflammation, sustain intestinal epithelial barrier integrity, and modulate behavior. We are using pre-clinical models and in vitro assays for these investigations with the goal of providing a mechanistic understanding of dairy food - probiotic interactions that benefit human health. Our findings support the hypothesis that probiotic efficacy depends on food delivery format. Our in vitro assessments indicate that dairy and probiotics work synergistically to benefit health.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Targeting the microbiome with prebiotics an probiotics to improve human health, WellNext Science Day
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Synergy between milk and probiotics for gastrointestinal health. Host Microbe Interactions annual UC Davis Research Retreat, October 2017