16th International Symposium on

Milk Genomics and Human Health

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Meet our Consortium-Level Sponsors
Arla Foods amba
S�nderh�j 14
8260 DK-Viby J
Denmark
Contact: Peter Langborg Wejse, Head of Open
Innovation, Universities & Consortia
Email: pewej@[Link]
Phone: +45 7217 7819
[Link]
Arla Foods is an international dairy company owned by 9,900 farmers from Denmark,
Sweden, the UK, Germany,
Belgium, Luxembourg and the Netherlands. Arla Foods is one of the leading players
in the international dairy
arena with well-known brands like Arla�, Lurpak�, Puck� and Castello�. Arla Foods
is focused on providing good
dairy nourishment from sustainable farming and operations and is also the world's
largest manufacturer of
organic dairy products.
California Dairy Research Foundation (CDRF)
501 G Street, Suite 203
Davis, CA 95616
Contact: Denise Mullinax, Executive Director
Email : mullinax@[Link]
Phone : 530-753-0681
[Link]
The California Dairy Research Foundation (CDRF) was created as a not-for-profit
research corporation in 1988 for
the purpose of promoting research and development activities that benefit
California�s dairy producers and
processors in the consumer marketplace. The organization is uniquely positioned to
act as a link between
industry and R&D institutions, and to lead the development of needed science-based
research and education
programs in critical areas. CDRF utilizes its independent structure to lead and
deliver the best-practice research
and science-based education programs to promote industry innovation and long-term
sustainability. Close
collaboration with associated industry organizations allows research outcomes to be
fully communicated and
utilized to maximize stakeholder benefits.
Danone Nutricia
Uppsalalaan 12
3584 CT Utrecht, The Netherlands
Contact: Bernd Stahl, [Link].
Director Human Milk Research & Analytical
Science
Email: [Link]@[Link]
Phone: +31 30 2095 000
[Link]
Developing high-quality and clinically proven nutritional solutions involves
bringing together an array of internal
teams and external partners from across a wide range of disciplines. Throughout the
innovation process, from
scientific insights through technology, our teams work collaboratively to develop
nutritional products that meet
patient and consumer needs.
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NZO
Benoordenhoutseweg 46
2596 BC The Hague, The Netherlands
Contact: Stephan Peters
Email: peters@[Link]
Phone: +31 79 3430 351
[Link]/en/
The Dutch Dairy Association (Nederlandse Zuivel Organisatie, NZO) is an association
of the Dutch dairy industry,
representing the interests of 13 dairy companies. Combined, these companies process
98% of the milk in The
Netherlands into a wide range of dairy products including beverages, infant formula
and children�s food, cheese
and desserts, and ingredients for the broader food industry and the pharmaceutical
sector. The NZO aims to
reinforce its members� abilities to produce and market their products. To reach
this goal, NZO can be found in
The Hague and in Brussels, and it also participates in public dialogue.
Meet our Symposium-Level Sponsors
Aarhus University
Blichers All� 20, PO Box 50
DK-8830 Tjele Denmark
Contact: Lotte Bach Larsen Ph.D.
Email: lbl@[Link]
Phone: +4587158049
[Link]
[Link]
Aarhus University (AU) was founded in 1928 and considered a top ten university
among universities founded
within the past 100 years. It has a long tradition of partnerships with some of the
world's best research
institutions and university networks. The university�s goal is to contribute
towards solving the complex global
challenges facing the world. The university therefore strives to combine the high
level of academic standards of
its researchers with collaboration across disciplinary boundaries to combine
research in new ways and solve
challenges in close contact with the world around us.
Arla Food for Health
Agro Food Park 19
Skejby, Denmark
Contact: Anne Louise M�rkbak, Director
Email: anmor@[Link]
Phone: +45 4160 7119
[Link]/company/arla-food-for-health/
Arla Food for Health was established in 2015 as a public private partnership
between Arla Foods amba, Arla Foods
Ingredients Group P/S, Aarhus University and University of Copenhagen with the
common vision of �Discovering
the health effects of dairy and dairy ingredients.� The Partnership is founded in
the spirit of exploring the
competences between the four stakeholders to carry out best-in-class dairy health
and nutrition research and
become a platform for knowledge sharing, training and education.
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Arla Foods Ingredients Group P/S (AFI)
S�nderh�j 10 - 12
8260 DK-Viby J Denmark
Contact: Henrik J Andersen
Senior Executive R&D Advisor
Email: hejan@[Link]
Phone: +45 2095 1241
[Link]
Arla Foods Ingredients is a leading whey ingredient supplier for infant nutrition,
baking, dairy, medical
applications, sports nutrition and affordable foods. Arla Food Ingredients is known
for having the industry�s
highest levels of quality, innovation and food safety. Our functional and
nutritional whey proteins, milk minerals,
lactose and whey permeate can promote nutrition, improve texture, prolong freshness
and increase yield for
industrial food products. Forty-five scientific staff and 22 food and dairy
technicians work to discover and
carefully document the effects of our functional and nutritional ingredients. At
the application level, some 40
dairy technicians, scientists and engineers, work in well-equipped pilot plants to
help our customers prepare
these discoveries for success on their production lines and in the marketplace.
Arla Foods Ingredients Group P/S
is a subsidiary of Arla Foods amba, a global dairy cooperative owned by dairy
farmers in six countries.
National Dairy Council
10255 West Higgins Road, Suite 900
Rosemont, IL 60018
Contact: Christopher J. Cifelli, PhD
Vice President, Nutrition Research
Email: [Link]@[Link]
Phone: 847-627-3282
[Link]
National Dairy Council (NDC), the nonprofit organization founded by dairy farmers
and funded by the national
dairy checkoff program, has been committed to research-based nutrition education
and communications since
its start in 1915. NDC is dedicated to bringing to life the dairy community�s
shared vision of a healthy, happy,
sustainable world � with science as its foundation. NDC�s staff of registered
dietitians, researchers, and nutrition
experts educate on dairy�s role as part of balanced eating plans, as well as inform
people about dairy�s
contributions to health and sustainable food systems. For more information, visit
[Link] and
[Link] and follow NDC on Facebook and Twitter.
RAW Milk Institute
7221 So. Jameson
Fresno, CA 93706 USA
Contact: Sarah Smith, Director-Secretary
Email: sarah@[Link]
Phone: 559-970-5581
[Link]
Founded in 2011, the Raw Milk Institute is a nonprofit organization that mentors
and trains farmers using high
standards, risk management practices and validation testing for the production of
raw milk for human
consumption. RAWMI supports research and farmer training using scientific
principles with a full recognition of
both the risks and the benefits of raw milk production and consumption.
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Riddet Institute
Massey University
Private Bag 11 222
Palmerston North 4442
Phone: +64 6 951 7295
Email: info@[Link]
[Link]
The Riddet Institute is a premier center for fundamental and strategic scientific
research and training a new
generation of science leaders. Its area of expertise is at the intersection of food
material science, novel food
processing, human nutrition and gastrointestinal biology. The Institute integrates
partner organizations and
disciplines, generating knowledge and capability that transforms the New Zealand
food industry. The Riddet
Institute�s environment for discovery and learning is fertile ground for its
students, who will be the leaders of a
prosperous food industry in the future and one that has the ability to develop
innovative, healthier foods.
LinkedIn: [Link]/company/riddet-institute; Twitter: @RiddetIns
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Annual Symposium Photography and Social Media Guidelines
Oral presentations provide attendees with the most up-to-date scientific findings
from the fields of lactation,
milk, and dairy. The IMGC established photography and social media guidelines in
order to encourage the
presentation of new and unpublished results.
Photography/Social Media Policy
? Photography and social media. Taking photographs of presentation slides at the
Annual IMGC Symposia
is at the discretion of the presenter/study author. Sharing photographs of
presentation slides via social
media is also at the discretion of the presenter/study author.
? Each presenter/study author will announce verbally and visually on a slide at the
beginning of their
presentation, whether he/she approves of photos being taken of his/her slides and
whether he/she approves
sharing on social media. Below are example slides presenters may use to communicate
whether photography/social media are prohibited or allowed.
? When allowed, attendees may take photographs during presentations provided that
the photographs are
for personal, non-commercial use and are not for publication or rebroadcast without
the express written
permission of the author. Be sure to cite the presenter�s/study author�s name with
any social media posts
that include his/her presentation slides.
? Please do not use flash photography as it can distract the presenters or members
of the audience.
Social Media
The IMGC encourages the use of social media, when following the above photography
and social media policies
and within embargo restrictions, to update followers on the latest news coming from
the IMGC.
To engage on social media be sure to use the official meeting hashtag,
#2019MILKGENOMICS, in all your posts.
You can also follow these channels for meeting updates:
? Twitter: @imgcmedia
? Linked-in: [Link]
PHOTOGRAPHY PROHIBITED
PLEASE DO NOT take photos of
this presentation.
PHOTOGRAPHY ALLOWED
YOU MAY take photos of this presentation.
Please be considerate of others in the session.
SOCIAL MEDIA ALLOWED
YOU MAY post this presentation
SOCIAL MEDIA PROHIBITED
PLEASE DO NOT post this presentation
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MILK AND HEALTH IN A SUSTAINABILITY PERSPECTIVE
Day 1 � Tuesday, November 12, 2019
8:00 a.m. Registration
Hot Topics � Sustainability
Chairman: Henrik J. Andersen, Arla Foods Ingredients, Aarhus, Denmark
9:00 a.m. Welcome: Where We Have Been, Where We Are Going
Bruce German, University of California-Davis, CA, USA
9:15 a.m. Invited Speaker: � Liter of Milk a Day � What are the Implications?
Troels Kristensen, Aarhus University, Tjele, Denmark
9:45 a.m. Keynote Speaker: The Role of Dairy in a Sustainable Healthy Diet:
Evidence from a Large
Australian Population Health Survey
Brad Ridoutt, CSIRO Agriculture, Clayton South, Australia
10:30 a.m. Coffee Break
11:00 a.m. Keynote Speaker: Food Systems for Health, Env and Enterprise: The
SUSFANS Project and Its
Findings
Monika Zurek, Oxford University, Oxford, United Kingdom
11:45 a.m. Sustainable Dairy Production and Its Exploitation into Healthy Dairy
Products
Nina Aagaard Poulsen, Aarhus University, Tjele, Denmark
12:15 p.m. Student Travel Award Recipient: Genetic Variants and Post-Translational
Modifications of
Milk Proteins in Relation to Non-Coagulating Milk
Kajsa Nilsson, Lund University, Lund, Sweden
12:35 p.m. Lunch
The Microbiome and the Metagenome
Chairman: Bruce German, University of California-Davis, CA, USA
1:30 p.m. Keynote Speaker: Microbiota Impact in the Early Stages of Life and the
Effects on
Health Later in Life
Christopher Stewart, Newcastle University, Newcastle upon Tyne, United Kingdom
2:15 p.m. Invited Speaker: The Composition of the Perinatal Intestinal Microbiota
in Cattle
Mikael Niku, University of Helsinki, Helsinki, Finland
2:45 p.m. Coffee Break
3:15 p.m. The Composition of the Postprandial Metabolome of Human Blood and Urine
Depends on the Human Genome and the Dairy Products Consumed
Guy Verg�res, Agroscope, Bern, Switzerland
3:45 p.m. Student Travel Award Recipient: Infant Commensal Complexed with Secretory
IgA Prevents Enteric Infection
Vanessa Dunne-Castagna, University of California-Davis, CA, USA
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4:05 p.m. Panel Discussion: Linking Sustainability to the Microbiome and Metagenome
Moderator: Bruce German, University of California-Davis
Panelists: Troels Kristensen, Aarhus University; Brad Ridoutt, CSIRO Agriculture;
Monika Zurek,
Oxford University; Mikael Niku, University of Helsinki; Christopher Stewart,
Newcastle University
4:30 p.m. Update from 2018�s Most Valuable Presentation: Human Milk
Oligosaccharides Directly
Enhance Gut Barrier Integrity to Alter Infections
Ishita Shah, University of California-Davis, CA, USA
4:50 p.m. Highlights from IMGC�s �SPLASH!� milk science update� and Upcoming Hot
Topics
Danielle Lemay, Western Human Nutrition Research Center, USDA, CA, USA
5:15-7:15 p.m. Poster Reception with Refreshments
6:15-8:15 p.m. Joint Scientific Advisory Council/Steering Committee Meeting (Closed
Session)
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Day 2 � Wednesday, November 13, 2019
Comparative Biology and Genomics
Chairman: Danielle Lemay, Western Human Nutrition Research Center, USDA, CA, USA
8:30 a.m. Invited Speaker: Composition, Structure and Digestive Dynamics of Milks
from
Different Species
Debashree Roy, Riddet Institute, Massey University, Palmerston North, New Zealand
9:15 a.m. Invited Speaker: Functionality and Structural Aspects of Milk Genetic
Variants
Jared Raynes, Food and Nutrition, Commonwealth Scientific and Industrial Research
Organization
(CSIRO), Werribee, Australia
9:45 a.m. Genomic Variation in Milk Proteins in European Cattle Breeds
Bernt Guldbrandtsen, Aarhus University, Tjele, Denmark
10:15 a.m. Coffee Break
10:45 a.m. Sequence-Based GWAS, Network and Pathway Analyses Reveal Genes Co-
Associated with
Milk Cheese-Making Properties and Milk Composition in Montb�liarde Cows
Marie-Pierre Sanchez, INRA, Jouy-en-Josas, France
11:15 p.m. Student Travel Award Recipient: Variability of Oligosaccharides in Milk
from Native
Nordic Cattle Breeds
Anne Vuholm Sunds, University of Aarhus, Tjele, Denmark
11:35 a.m.-12.30 p.m. Lunch
Inherent Milk Characteristics and Functionalities
Chairman: Marie Paulsson, Lund University, Lund, Sweden
12:30 p.m. Keynote Speaker: Casein Micelle Mineralization
Thom Huppertz, Friesland Campina, The Netherlands
1:15 p.m. Invited Speaker: Milk Cathepsins � Implications for Health and
Functionality
Lotte Bach Larsen, Aarhus University, Tjele, Denmark
1:45 p.m. Student Travel Award Recipient: High Proportion of Transient Neonatal
Zinc Deficiency
(TNZD) Causing Alleles in the General Population
Yarden Golan, Technion-Israel Institute of Technology, Haifa, Israel
2:05 p.m. Coffee Break
2:30 p.m. Invited Speaker: Understanding the Function of the Milk Fat Globule
Membrane
Carolyn M. Slupsky, University of California-Davis, CA, USA
3:00 p.m. Isolation, Characterization, and Biological Properties of Milk
Extracellular Vesicles
Jan Trige Rasmussen, Aarhus University, Aarhus, Denmark
3:30 p.m. Invited Speaker: The Role of Whey Protein in Postprandial Glycaemic
Control
Daniela Jakubowicz, Tel Aviv University, Israel
4:00 p.m. Natural Glycosidases in Bovine Milk
S�ren Drud-Heydary Nielsen, Aarhus University, Tjele, Denmark
4:30 p.m. End of day 2
6:30-10:30 p.m. Symposium Group Dinner at Smedjen (The Smitty), Centralv�rkstedet
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Day 3 � Thursday, November 14, 2019
Matrix Effects of Milk and Milk Components
Chairman: Peter Langborg Wejse, Arla Foods, Aarhus, Denmark
9:00 a.m. Keynote Speaker: The Dairy Matrix and Health: Current Evidence and
Knowledge Gaps
Michelle McKinley, Queens University Belfast, Belfast, United Kingdom
9:45 a.m. Invited Speaker: Dairy Matrix Effects: Response to Consumption of Dairy
Fat Differs when
Eaten within the Cheese Matrix � a Randomized Controlled Trial
Emma Feeney, University College Dublin, Dublin, Ireland
10:15 a.m. Coffee Break
10:45 a.m. Discovering Added-Value Compounds in Dairy Streams: Proteomic Profiling
Reveals the
Presence of Xanthine Oxidase with Remarkable Antimicrobial Activity
Gulustan Ozturk, University of California-Davis, CA, USA
Being presented by Bruce German, University of California-Davis, CA, USA
11:15 a.m. Student Travel Award Recipient: Maternal Allergy and the Presence of
Non-Human
Proteinaceous Material in Human Milk
Pieter Dekker, Wageningen University, Wageningen, The Netherlands
11:35 a.m. Panel Discussion: The Dairy Matrix
Moderator: Merete Myrup, Danish Agriculture & Food Council
Panelists: Michelle McKinley, Queens University Belfast; Emma Feeney, University
College Dublin;
Bruce German, University of California-Davis
12:00 p.m. Closing Remarks
12:15 p.m. Lunch
12:45 p.m. End of Conference
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Meet the Speakers
Pieter Dekker
Student Travel Award Recipient
Ph.D. Student
Wageningen University & Research
Bornse Weilanden 9
6700 AA Wageningen, p.o. box 17
Wageningen, The Netherlands
Phone: +31614787773
Email: [Link]@[Link]
Pieter Dekker studied Analytical Chemistry (BASc) and Food Technology ([Link].) with
a specialization in
Ingredient Functionality. Dekker started his Ph.D. research in 2018 at Wageningen
University & Research in the
department of Food Quality and Design. Under supervision of Kasper Hettinga, he is
investigating the human
milk protein and peptide profile, and the relation of this with maternal allergy
and allergy development in the
offspring.
Vanessa Dunne-Castagna
Student Travel Award Recipient
Graduate Student Researcher
University of California-Davis
One Shields Ave.
Davis, CA, 95616 USA
Phone: 916-743-4080
Email: vpdunnecastagna@[Link]
Vanessa Dunne-Castagna is a fourth-year graduate student in nutritional biology,
studying under the
mentorship of both Dr. Bo Lonnerdal and Dr. David A. Mills. Her interests include
understanding the complex
relationship between bioactive milk components and the development of the infant
gut microbiome.
Specifically, Dunne-Castagna has been investigating milk secretory immunoglobulin A
and its dynamic interplay
between the host, commensals and pathogens. She is also a community college biology
instructor teaching
predominately pre-nursing students. She enjoys bringing the latest research on
host-microbial interactions into
the classroom to better equip future health care workers.
Emma Feeney, Ph.D.
Assistant Professor
Institute of Food and Health
University College Dublin
2.16a Science Centre South
Belfield, Dublin 4, Ireland
Phone: 01-7162247
Email: [Link]@[Link]
Emma Feeney is an Assistant Professor at University College Dublin�s Institute of
Food and Health. Formerly the
research manager at Food for Health Ireland (FHI), a dairy functional foods
research center, Feeny�s research
interests span various aspects of dairy foods, from dairy food consumption patterns
and their contribution to
health, to food structure and the impact on digestion, absorption and assimilation
of nutrients. Sensory
evaluation and variation in sensory perception are also areas of significant
interest.
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Meet the Speakers
J. Bruce German, Ph.D.
Director, Foods for Health Institute
Professor, Department of Food Science & Technology
University of California, Davis
One Shields Ave.
Davis, CA 95616 USA
Phone: +1-530-752-1486
E-mail: jbgerman@[Link]
Bruce German received his Ph.D. from Cornell University, joined the faculty at the
University of California, Davis
in 1988, and is currently Director of the Foods for Health Institute and professor,
at University of California,
Davis. His research interests include the structure and function of dietary lipids,
the role of milk components in
food and health, and the application of metabolic assessment to personalizing diet
and health.
Yarden Golan
Student Travel Award Recipient
Ph.D. Student
Professor Assaraf's Lab, The Faculty of Biology
Technion - Israel Institute of Technology
Haifa 32000, Israel
Phone: 972524877011
Email: golanedray@[Link]
Yarden Golan completed a bachelor of Nutrition Science and later started to study
for a master�s degree under
the supervision of Professor Assaraf at the Technion (Israel). After two years,
Yarden shifted to the direct Ph.D.
track. Her Ph.D. research focused on the transport mechanism of zinc into human
breast milk. She will soon
start her post-doctoral fellowship under the mentorship of ProfessorNadav Ahituv
from UCSF, who is an expert
in genomics analyses.
Bernt Guldbrandtsen, Ph.D.
Associate Professor
Center for Quantitative Genetics and Genomics
Aarhus University
AU Foulum, Box 50
8830 Tjele, Denmark
Phone: (45)31177093
Email: [Link]@[Link]
Bernt Guldbrandtsen does research in animal genomics and quantitative genetics and
breeding of farm animals,
primarily in dairy cattle. In recent years, he has been involved with the 1000 Bull
Genomes Project. He trained
as a biologist and earned a Ph.D. in theoretical population genetics at Aarhus
University in Denmark. Since 2007,
he has held a permanent position at Aarhus University�s Center for Quantitative
Genetics and Genomics.
Photo
Not
Available
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Meet the Speakers
Thom Huppertz, Ph.D.
Principal Scientist Food Structuring
FrieslandCampina Innovation Centre
Bronland 20
6708 WH
Wageningen, The Netherlands
Phone: +31 6 11187512
Email: [Link]@[Link]
Thom Huppertz hold an [Link]. from Wageningen University and a Ph.D. from University
College Cork. His
research career includes academic and industrial research in the field of dairy
science and technology, with
primary focus on milk protein and their functionality, from biosynthesis to
digestion in the human body. He
currently combines the roles of Professor of Dairy Science and Technology at
Wageningen University, Principal
Scientist at FrieslandCampina, Visiting Professor at Victoria University, and
Editor-in-Chief of International Dairy
Journal.
Daniela Jakubowicz, M.D.
Professor
Diabetes Unit, Wolfson Medical Center
Tel Aviv University
Herbert Samuel, 36 St.
Tel Aviv Israel 58100
Phone: 972 5 0810 5552
Email: [Link]@[Link]
Clinical Professor of Medicine at Virginia Commonwealth University, USA and in the
Diabetes Unit, at E.
Wolfson Medical Center, Tel Aviv University, Israel.
Daniela Jakubowicz�s large scientific contribution, over 25 years at Virginia
Commonwealth University and since
2009 in Israel in the Diabetes Unit Wolfson Medical Center, is focused on the
effect of meal timing and
composition on circadian clock gene regulation of glucose metabolism, insulin
resistance, obesity, polycystic
ovary and type 2 diabetes. Her studies are published in the most important journals
including three seminal
papers in the New England Journal of Medicine.
Troels Kristensen, Ph.D.
Senior Scientist
Department of Agroecology
Aarhus University, Denmark
Research Center Foulum
P.O. Box 50
DK 8830 Tjele, Denmark
Phone: +45 26 12 55 83
Email: [Link]@[Link]
Troels Kristensen holds a Ph.D. in animal science and is presently senior scientist
at Department of Agroecology,
Aarhus University. Farming systems research has been his main research method to
investigate different aspects
of livestock production, management and the impact of farming practice on the
environment. Troels is member of
the climate advisory board and the research board for Danish Cattle Federation.
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Meet the Speakers
Lotte Bach Larsen, Ph.D.
Professor
Department of Food Science
Aarhus University
Blichers All� 20, PO Box 50
DK-8830 Tjele Denmark
Phone: +45 87158049
Mobile: +45 22819282
Email: lbl@[Link]
Lotte Bach Larsen is a professor at Department of Food Science, University of
Aarhus, Denmark. She leads an
active science team in �Food Chemistry and Technology,� and has published over 120
international, peerreviewed articles, in addition to book chapters. Bach Larsen
works within properties and molecular structures
of milk proteins and milk composition, including proteolysis. She is using
proteomic and peptidomic methods
for analyses and profiling of food-based proteins and peptides, protein
modifications, natural and process
induced. Member of Local Organizing Committee for International Symposium on Milk
Genomics and Human
Health in Aarhus (2014 and 2019), and a recipient, together with the Danish team
behind the Milk Genomics
Initiative in Denmark, of the Milk Prize awarded by the Danish Dairy Board in 2017
in acknowledgement of its
contribution to the basic understanding of the relations between cow genes and milk
fingerprint and its
resulting properties.
Danielle G. Lemay, Ph.D.
Research Scientist
USDA ARS Western Human Nutrition Research Center
430 West Health Sciences Drive
Davis, CA 95616 USA
Phone: 530-752-4748
E-mail: [Link]@[Link]
Danielle Lemay is a Research Scientist at the U.S. Department of Agriculture�s
Western Human Nutrition
Research Center in Davis, California. She is also an Associate Adjunct Professor
with the Dept. of Nutrition and
faculty member of the Genome Center at the University of California-Davis. She is
the Founder and Executive
Editor of �SPLASH! milk science update,� the official e-newsletter of the
International Milk Genomics
Consortium (IMGC) for which she has selected and reviewed over 350 lay articles on
milk science. She has been
a member of the IMGC�s Scientific Advisory Committee since 2007. Lemay�s research
program was previously
focused on the genetics of milk production, mammary biology, and milk-oriented
microbes. At the USDA, her
lab�s focus is the effect of diet on gut microbes and gastrointestinal health. She
has a Ph.D. and MS in Nutritional
Biology from UC Davis, and a BS in Electrical Engineering & Computer Science from
MIT.
Michelle McKinley, Ph.D.
Professor
Queens University Belfast
School of Medicine, Dentistry & Biomedical Sciences
Grosvenor Road
Belfast BT12 6BJ, United Kingdom
Phone: 00 44 2890 97 8936
Email: [Link]@[Link]
Michelle McKinley is a Professor of Nutrition at the Centre for Public Health,
School of Medicine, Dentistry and
Biomedical Sciences, Queen's University Belfast. Professor McKinley�s research
investigates the ability of
dietary interventions to modify nutritional status and risk of chronic disease,
particularly diabetes and
cardiovascular disease, as well as exploring novel approaches to encouraging and
supporting diet and lifestyle
behavior change and weight management throughout the life course.
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Meet the Speakers
Merete Myrup, Ph.D.
Director of Dairy Nutrition
Danish Agriculture & Food Council
Agro Food Park 13
DK 8200 Aarhus N
Phone: +45 3339 4818
Email: mmc@[Link]
As the Director of Dairy Nutrition, Merete Myrup focuses on research and
communication on the nutritional
benefits of dairy products, where communication is towards both health care
professionals and consumers.
She is also engaged in both European and international collaboration groups and
projects, i.e. the European
Milk Forum, where the dairy matrix is a top priority. In recent years she has
extended her focus to also include
sustainability, with a focus on the role of dairy products in sustainable diets.
S�ren Drud-Heydary Nielsen, Ph.D.
Postdoc
Department of Food Science
Aarhus University
Blichers All� 20,
Postboks 50
DK-8830 Tjele, Denmark
Phone: +45 871 58062
Email: sodn@[Link]
The overall aims of S�ren Drud-Nielsen�s research are to investigate the chemical
and nutritional properties of
proteins in food. He is determining the extent of natural and process-induced post-
translation modifications
(PTMs) on proteins, mapping the digestion of proteins in the human gut and
investigating bioactivities of these
digests on human health. He is developing bioinformatic tools and analytical
methods using LC-MS (scan and
multiple reaction monitoring mode) to identify and quantify proteins, peptides and
their PTMs.
Mikael Niku, Ph.D.
Senior Lecturer
University of Helsinki, Veterinary Biosciences
PO Box 66
00014 Helsinki, Finland
Phone: +358 50 5719159
E-mail: [Link]@[Link]
Mikael Niku is a senior lecturer at Veterinary Biosciences, University of Helsinki.
His research currently focuses
on very early host-microbiota interactions: How does the maternal microbiota affect
the development of the
immune system in the offspring? He is primarily interested in large domestic
mammals, especially cattle. His
research interests also include the epigenetic regulation of cellular
differentiation.
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Meet the Speakers
Kajsa Nilsson
Student Travel Award Recipient
Ph.D. Student
Lund University
Department of Food Technology, Engineering and Nutrition
Box 124
221 00 Lund, Sweden
Phone: +46 739 36 41 70
Email: [Link]@[Link]
Kajsa Nilsson is currently a Ph.D. student at Department of Food Technology,
Engineering and Nutrition, Lund
University with project title �Non-coagulating milk from Swedish Red Dairy Cattle.�
She will defend in May 2020
and then it is time for new adventures.
Gulustan Ozturk, Ph.D.
Postdoctoral Researcher
Department of Food Science & Technology
University of California, Davis
Robert Mondavi Institute, South
Davis, CA 95616, USA
Phone: 530-574-7766
Email: gozturk@[Link]
Gulustan Ozturk received her Ph.D. at the University of California, Davis where her
research focused on the
virulence mechanisms of the foodborne pathogen Yersinia enterocolitica and its
survival and transmission in the
environment. She is currently a postdoctoral researcher at the University of
California, Davis where her current
research is focused on human and bovine milk-based antimicrobials to lead to
industrial applications in support
of healthy microbial communities. Her current work also focuses on isolation and
characterization of bioactive
compounds in dairy products and industrial side streams.
Nina Aagaard Poulsen, Ph.D.
Associate Professor
Department of Food Science, Aarhus University
Blichers All� 20
8830 Tjele, Denmark
Phone: +45 87157997
Email: [Link]@[Link]
Nina Aagaard Poulsen obtained her Ph.D. degree in conservation genetics from
Department of Biological
Sciences, Aarhus University, Denmark in 2008. She has worked at the Department of
Food Science, Aarhus
University since 2009. Her research focuses on understanding factors affecting
variation in composition of
bovine milk especially in relation to genetic background, feeding and sustainable
production and to explore
how such variations affect nutritional, technological and health-related properties
of milk, which ultimately can
be exploited for milk differentiation.
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Meet the Speakers
Jan Trige Rasmussen, Ph.D.
Senior Scientist
Dept. of Molecular Biology and Genetics
Aarhus University
Gustav Wieds Vej 10C, 3133.221
8000 Aarhus C, Denmark
Phone: +45 87 15 54 62
E-mail: jatr@[Link]
Jan Trige Rasmussen earned a Ph.D. in basic intracellular lipid metabolism at the
University of Southern
Denmark and took a position at Aarhus University (AU). His researcher findings are
presented in peer-reviewed
multidisciplinary papers focusing on isolation, characterization and bio-assaying
of new and less known
proteins, mostly from milk. The affiliated section for Molecular Nutrition is one
of the major centers in Denmark
for basic milk component research.
Jared Raynes, Ph.D.
Research Scientist
Commonwealth Scientific and Industrial Research Organisation (CSIRO)
671 Sneydes Rd
Werribee, Victoria, 3030 Australia
Phone: +61 3 9731 3358
Email: [Link]@[Link]
Jared Raynes is a research scientist at the CSIRO in Melbourne, Australia in the
Food Structure group where he
has been since 2012. His research areas include the use of advanced biophysical
techniques such as small angle
X-ray and neutron scattering to investigate the casein micelles of a variety of
mammals, understanding the
genetic variation of milk proteins, the use of genetic selection as a means to
manufacture unique dairy products
and most recently using protein structuring to manipulate the digestion of foods
and manufacture unique
nanocarriers of bioactive compounds.
Brad Ridoutt, Ph.D.
Principal Research Scientist
CSIRO Agriculture
Bayview Avenue (Private Bag10)
Clayton South, Vic 3169, Australia
Phone: +61 3 9545 2159
Email: [Link]@[Link]
Brad Ridoutt is a Principal Research Scientist with Australia�s national science
agency � The Commonwealth
Scientific and Industrial Research Organisation (CSIRO). His expertise is in life
cycle sustainability assessment in
the agriculture and food sectors which is used to address strategic challenges in
relation to climate change,
water scarcity, sustainable food systems, and sustainable diets. Ridoutt is engaged
in a range of international
processes relating to the standardization of sustainability assessment and
environmental labelling.
20 | Page
Meet the Speakers
Debashree Roy
Ph.D. Student
School of Food and Advanced Technology
Massey University
Private Bag 11222
Palmerston North, New Zealand
Phone: +64 221847906
E-mail: [Link]@[Link]
Debashree Roy is a final year Ph.D. student with Riddet Institute and Massey
University in New Zealand. She
graduated with a master�s degree in food technology in 2013 and began her research
career the same year with
IFFCO, leading the research and development for their agri-business group. Her
current research focus is in the
area of food structure, digestion, and health. She aims to understand how the
microstructural and
physicochemical changes in dairy food during digestion influences its health
outcomes.
Marie-Pierre Sanchez, Ph.D.
Engineer
INRA
GABI Demaine de Vilvert
78350 Jouy-en-Josas, France
Phone: +33 13 46 52 18
Email: [Link]@[Link]
Marie-Pierre Sanchez, is a quantitative geneticist by training and has been an
engineer at INRA-GABI in France
since 2000. She first worked in pig genetics and joined the Bovine Genetics and
Genomics team (G2B) in 2012.
She just finished a Ph.D. on �Genetic analysis of milk protein composition and
cheese-making properties
predicted from MIR spectra.� She has a broad experience in genome-wide association
studies. She has
published 29 scientific articles (h-index = 13).
Ishita Shah, Ph.D.
2018�s Most Valuable Presentation
Associate Project Scientist
Department of Food Science & Technology
University of California Davis
1221 Robert Mondavi Institute, South
Davis, CA 95616USA
Phone: 530-754-7962
Email: imshah@[Link]
Ishita Shah received her Ph.D. at UMBC where she discovered a new mechanism of
transcription activation
in bacteria. Her postdoctoral work at Columbia University led to the identification
of bacterial cell wall
fragments causing an exit from dormancy. At Genentech, she identified and validated
targets in pathogenic
bacteria in a drug discovery program. At the Foods for Health Institute, UC Davis,
Shah works towards the
identification of specific milk bioactives that alter pathogenesis.
21 | Page
Meet the Speakers
Carolyn M. Slupsky, Ph.D.
Professor
Departments of Nutrition and Food Science & Technology
University of California, Davis
One Shields Ave.
Davis, CA 95616 USA
Phone: 530-752-6804
E-mail: cslupsky@[Link]
Carolyn Slupsky holds a joint appointment in the Departments of Nutrition and Food
Science & Technology at
the University of California, Davis, and is the Chair of the Graduate Group in
Nutritional Biology. She was named
a Chancellor�s Fellow in 2015, and the Kinsella Endowed Chair in Food, Nutrition,
and Health in 2016. Her lab is
interested in the nexus between food, the gut microbiome, and health, with a focus
on infants and young
children.
Christopher Stewart, Ph.D.
Research Fellow and Group Leader
Newcastle University Medical School
Framlington Place
NE2 4HH United Kingdom
Phone: +447986983003
Email: [Link]@[Link]
Christopher Stewart has researched the early life microbiome in health and disease
for the past decade,
specializing in infants born premature (<32 weeks gestation). In that time, he has
published over 50 peerreviewed manuscripts, including two recent articles in
Nature. His research group performs both
computational and wet-lab experimentation focused on microbial-host interaction in
the gut.
Anne Vuholm Sunds
Student Travel Award Recipient
Ph.D. Student
Department of Food Science
Aarhus University
Blichers All� 20, Postboks 50
8830 Tjele Denmark
Phone: +45 87157802
Email: annesunds@[Link]
Anne Vuholm Sunds is a Ph.D. student at Department of Food Science, Aarhus
University, Denmark. Her
research focuses on understanding the variations in glycosidases naturally present
in milk, and their potential
effect on free oligosaccharides and glycoconjugates during processing. In September
2018 she joined the Barile
lab at the University of California, Davis, where she investigated the variation
and content of oligosaccharides
in milk from a unique sample set of eight native Nordic dairy cattle breeds.
22 | Page
Meet the Speakers
Guy Verg�res, Ph.D.
Head of the Functional Nutritional Biology Group
Agroscope
Schwarzenburgstrasse 161
3003 Bern, Switzerland
Phone: +41 (0)58 463 81 54
Email: [Link]@[Link]
Guy Verg�res is head of the Functional Nutritional Biology Group and deputy head of
the Food Microbial
Systems Research Division at Agroscope, Federal Department of Economic Affairs,
Education and Research in
Bern Switzerland. He is also a lecturer on nutrigenomics at ETH-Zurich and the
University of Lausanne and a
member of the Management Board of the nutrigenomics organization NuGO. The
translational research of
Verg�res focuses on the impact of food fermentation, using milk as a model food, on
human metabolism,
making use of the recent advances in analytical strategies in food (foodomics) and
nutrition (nutrigenomics)
sciences.
Monika Zurek, Ph.D.
Senior Researcher
Environmental Change Institute
Oxford University
South Parks Road
Oxford OX1 QY, United Kingdom
Phone: +44 7824024650
Email: [Link]@[Link]
Monika Zurek is a senior researcher at the Environmental Change Institute at the
University of Oxford and a
member of the ECI Food Systems Group. For more than 15 years she has worked on food
systems, environment
and development interactions in research (CIMMYT, ECI) and international
organizations (FAO) as well as in the
consulting (Climate Focus BV) and the philanthropic sectors (B&M Gates Foundation).
Presentation Abstracts
24 | Page
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Presentation Abstracts
9:15 a.m. Invited Speaker: � Liter of Milk a Day � What are the Implications
Troels Kristensen, Aarhus University, Tjele, Denmark
Agriculture and food consumption contribute to emission of greenhouse gasses and
the
pressure on global land use and biodiversity. It is generally agreed that livestock
products and
especially foods from cattle are related to a high emission of ammonia (NH3) and
greenhouse
gasses, mainly CH4 and N2O, as well as a large land use. This, together with an
estimated increase
of 1-2% in annual demand for milk and milk products, due to the increasing global
population
and expected change in consumption, calls for a better understanding of how dairy
farming can
develop over time to meet the global environmental challenges and at the same time
meet the
demand from the consumers.
Dairy farming across the world is very diverse in terms of size, productivity,
feeding and
management and by that the impact on the environment. Dairy farming is a multiple
production
system that besides milk as the main product, produces meat from the replacement
cows,
surplus heifers and the male calves. The herd structure, reproduction strategy and
use of
different breeding technologies � like sexed semen or crossbreeding � can change
the
proportion of milk and meat. Livestock also produce manure, which can replace
artificial
fertilizer, applied either directly in the fields during grazing or after
collection. Further, new
technologies like acidification or utilization for biogas can increase the
fertilizer value and
reduce the environmental impact related to emission of nitrogen (N) and methane.
Cattle,
being ruminants, have the ability to change feed from plants high in fiber content
to human
edible products and utilize low value protein as well. High fiber feed can be the
entire plant, like
grass, or as waste products coming from bio refining of several kind of biomass,
where
components relevant for monogastric animals or even human are extracted as high
value
products leaving the fiber components for feeding of cattle. This can contribute to
a change
towards plant productions system with a higher capacity of storage of carbon in the
soil �
carbon sequestration � and thus reduce the climate impact.
In the end, the environmental impact of the food produced is the sum of emissions,
direct or
indirectly, related to all the activities in the chain. To compare multiple system
the sum needs
to be allocated to the different products in order to compare the sustainability of
the product
and of the system taking into account the change in food production and the related
consumption. In order to understand some of the interactions between production and
environmental impact this paper takes the starting point from the Danish production
of milk
with the aim to illustrate how changes through the total chain from fed production,
feeding
and management at the dairy farm to consumer choice affects the impact on climate
and other
sustainability indicators.
9:45 a.m. Keynote Speaker: The Role of Dairy in a Sustainable Healthy Diet:
Evidence from a Large
Australian Population Health Survey
Brad Ridoutt, CSIRO Agriculture, Clayton South, Australia
The recent IPCC Special Report on climate change and land has again focused
attention on the
subject of sustainable diets and the need for urgent adoption of lower greenhouse
gas (GHG)
emission dietary patterns. The food system is an important source of these
emissions, as well
as other important environmental impacts. However, it is difficult to generalize
about dietary
strategies to achieve healthy diets with lower environmental impacts because food
systems
vary in different regions, as do the local food cultures, local public health
nutrition challenges,
and the importance of individual foods in relation to under-consumed nutrients by
specific
26 | Page
Presentation Abstracts
population subgroups. In addition, most opportunities to intervene in food systems
or in public
health nutrition are at the local or national scales. This underscores the need for
context specific
evaluation and any generalized strategy or suggestion about global reference diets
should be
treated with skepticism. In the case of dairy products, there is presently
contradictory advice
about the level of intake that is appropriate in a lower GHG emission healthy diet.
In Australia,
a country with a long cultural tradition of dairy product consumption, the
Australian Dietary
Guidelines recommend that most Australians should consume more reduced fat milk,
yogurt
and cheese. On the other hand, a range of studies, mainly involving scenario
modelling, have
indicated that dairy product intake should be reduced or avoided.
Here, results are presented from a collection of studies involving a dataset of
more than 9,000
adult daily diets obtained from the Australian Health Survey. For each individual
diet, carbon
and water-scarcity footprints were quantified, along with a diet quality score
expressing level
of conformance with the food group-based Australian Dietary Guidelines. These
studies show
that the factor of overwhelming importance in differentiating higher quality diets
with lower
environmental footprint from lower quality diets with higher environmental
footprint is the
intake of non-core (or discretionary) foods. According to the Australian Dietary
Guidelines,
these foods should be eaten only occasionally and in small quantities. As well as
inflating dietary
environmental footprints, the excessive consumption of discretionary food is linked
to excess
energy intake and the escalating rates of overweight, obesity and diet-related
chronic disease
which are major public health nutrition concerns in Australia. Generally,
Australians should
consume less discretionary food and more nutrient rich core foods, including dairy
products.
In addition, a subgroup of higher diet quality and lower GHG emission (HQLE) diets
has been
studied (N=1732). This subgroup had a diet quality score 37% above and GHG
emissions 43%
below the population average. It was found that core dairy products were a very
common
constituent, as 90% included milk, cheese or yogurt. The HQLE subgroup was
subsequently
divided into tertiles according to core dairy product intake and nutrient profiling
was
undertaken. Within the HQLE subgroup, higher levels of core dairy intake were
associated with
significantly higher attainment of adequate intake of a broad range of individual
nutrients. For
example, only 5.4% and 21.4% of diets in the low and medium dairy intake tertiles
achieved the
recommended intake of calcium. This compared to 73.9% for the higher dairy intake
tertile. The
HQLE subgroup of daily diets is important because it reflects the actual food
patterns of
Australians that already have more desirable dietary characteristics. Our results
show that core
dairy products are a common constituent of lower GHG emission healthy diets and
that high
levels of dairy intake are not inconsistent with a lower GHG emission diet, but
rather critical to
achieving adequate nutrition.
11:00 a.m. Keynote Speaker: Food Systems for Health, Env and Enterprise: The
SUSFANS Project and It�s
Findings
Monika Zurek, Environmental Change Institute, University of Oxford, Oxford, United
Kingdom
Food systems around the world will have to transform to achieve what has been
recently coined
�Sustainable Food and Nutrition Security�. Food systems are complex webs of actors
(e.g.
farmers, processors, retailers, transporters, consumers, etc.) and their
activities, the drivers of
their decisions and their various outcomes and implications for societal
objectives. For the EU
food system, the outcomes that its various actors are interested in have been
described as:
Balanced and sufficient diets for EU citizens, Reduced environmental impacts,
Viability of agrifood businesses, and Equitable outcomes and conditions of food
actors. The H2020 research
project SUSFANS (Metrics, models and foresight for sustainable food and nutrition
security in
the EU, [Link]) has assessed the sustainability performance of the EU food
system,
and created foresight on future food production, diets and sustainability impacts
across the
four societal goals. For that the project created a participatory process to map
the food system
27 | Page
 Presentation Abstracts
and its actors and develop a set of performance metrics to assess and visualize
system�s
performance and possible trade-offs associated with innovation options for food
system
transformation. The SUSFANS Visualizer gives insight into the sustainability
performance of
European diets and food systems for the years 2010 to 2050. In this way one can
compare the
impact of strategies and policies and adjust as needed. The interactive tool also
shows where
measurable policy targets and/or data are missing, such as on equity and social
justice in the
food system. Results indicate that the EU food system is not �future-proof� enough,
and its
sustainability performance is weak. Current nutritional patterns are in imbalance,
their
environmental impact is too large and economic viability and social justice are
under pressure.
In order to achieve sustainable diets and a sustainable food supply system in the
EU by 2030, a
rapid transformation of food production, consumption and trade is required. Finding
suggest
that there is a need to develop an EU policy protocol to monitor the health and
sustainability
impact of food consumption and intake; to better enable market decisions that
support a
transformation for sustainable and healthier food supply and consumption; to
develop
intervention strategies that involve experimentation, market intervention and
social innovation
and combine instruments with impact on consumption and production and maintain them
long
term; to reconnect different policies, under an aligned multi-level and multi-
dimensional food
policy framework in the EU and Member States.
11:45 a.m. Sustainable Dairy Production and its Exploitation into Healthy Dairy
Products
Nina Aagaard Poulsen, Aarhus University, Tjele, Denmark
Reducing climate footprints and making milk production more sustainable is an
urgent need
for future dairying. Increasing yields per cow will lead to lower carbon footprints
per kg of
milk produced, but still it is of importance to secure healthy animals and a
premium milk
quality. Mitigating methane emission through genetic selection and specific
feedings are at
play, but also new management strategies, as e.g. extended lactation may be a valid
alternative. By delaying insemination, longer calving intervals are achieved, which
can be a
beneficial strategy for high yielding dairy cows due to a reduction in feed use per
kg milk
produced and fewer risk periods around calving. However, there are still concerns,
whether
extended lactation for more than the traditional 12 months impairs milk quality. In
two different
studies, we have documented that up to 18 months calving interval does not impair
the milk
quality and that gestation stage expressed as days to next calving rather than
lactation length
affects the milk quality including protein-related traits and cheese making
properties.
Another important aspect of sustainable dairy production is the interplay between
the
metagenome, genome and methane emission on one hand and milk quality on the other
hand,
which is not well understood. In a recent study, we have collected milk and rumen
samples from
340 Holstein cows and documented that the amount of variance explained by the rumen
bacteria was generally lower than the genetic component, when it comes to milk
fatty acids.
However, the rumen microbiome had a pronounced influence on the content of odd
chain, as
well as polyunsaturated C18 fatty acids in the milk, and to a lesser extent on the
content on the
short and medium chain fatty acids in the milk. Thus, manipulation of the rumen
microbiome
may affect milk quality. As vitamin B12 and B2 in milk are naturally synthesized by
the rumen
microbiome, they may be prone to changes in the rumen microbiome. Both vitamins
display
significant variation in milk from Danish Holstein cows, but to which extent this
relates to the
microbiome is currently not explored, but this association will be explored in near
future. A new
project studies nitrogen (N) and phosphorous (P) metabolism through controlled
feeding
experiments and evaluates N and P contents in feed, in excretions, as well as their
incorporation
into milk components. Regulations demand a better monitoring of the N and P flow
from farms
to the environment, but from a milk perspective changes in N/P supplementation may
also
affect the protein-mineral balance in milk, and thereby affect milk functionality.
We have
28 | Page
Presentation Abstracts
conducted two feeding trials (incomplete Latin square experimental design n=24 and
n=36,
respectively) specifically addressing N and P efficiencies and their associated
effect on milk
quality, including urea and P shares in milk. Our primary aim is to secure that
milk quality is not
compromised in the search for more sustainable solutions for dairy production
systems.
12:15 p.m. Student Travel Award Recipient: Genetic Variants and Posttranslational
Modifications of
Milk Proteins in Relation to Non-Coagulating Milk
Kajsa Nilsson, Lund University, Lund, Sweden
The ability of milk to coagulate after addition of rennet is a crucial step in
cheese production.
We have previously shown that Swedish Red Dairy Cattle (RDC) has a high prevalence
of
individuals, 18%, that produce non-coagulating (NC) milk. This has a negative
effect on cheese
yield, and thus it is of high interest for dairies to eliminate NC milk. Previous
studies have tried
to link NC milk to different genetic variants of milk proteins and other studies
have linked
posttranslational modifications (PTM) of milk proteins to coagulation ability of
milk. One type
of PTM is glycosylation of ?-casein, but the results from earlier studies are
contradictory, since
it has been found that glycosylation of ?-casein could both increase or decrease
the rennetcoagulation ability of milk. The aim of this study was to determine the
protein profile in milk
from Swedish RDC, including PTMs of the caseins, in order to explore links between
the milk
and NC milk. Additionally, the results from this study can be used to investigate
the overall
protein profile of Swedish RDC cows and compare with other breeds.
Milk samples from 619 individual Swedish RDC cows were analyzed using liquid
chromatography-high resolution mass spectrometry and the obtained masses were
compared
to Uniprot database to determine genetic variants of both casein and whey proteins.
PTMs
were determined by using masses, where phosphorylations of the genetic variants of
casein, as
well as glycosylations of ?-casein, were obtained. Both the number of
glycosylations as well as
the glycosylation variant (A-E) were determined. The genetic variants and PTMs were
combined
with coagulation parameters measured in the milk samples, in order to distinguish
the protein
profile of NC milk from Swedish RDC. In addition, composition and physical
properties of the
milk samples were used in the study to fully explore the pattern for NC milk.
Genetic variants of milk proteins can be determined both genetically and
phenotypically, and
by measuring the variants in milk samples, the results will show which variants are
expressed at
protein level in the milk. Thus far, we have found that ?-casein variant E, which
previously has
been shown to have a negative impact on rennet-induced coagulation, has decreased
in the
Swedish RDC breed. In our data set, we have also found at least six different
phosphorylation
isoforms of aS2-casein, which is very uncommon. Further analyses should reveal
additional
insights into the relations between detailed milk protein profile and NC milk
within Swedish
RDC. By gaining more knowledge of genetic variants and PTMs of milk proteins
affecting NC
milk, this can be used in the breeding strategy of Swedish RDC in order to reduce
the frequency
of NC milk, and also to get a better understanding of the mechanisms behind the
inability of
milk to coagulate.
1:30 p.m. Keynote Speaker: Microbiota Impact in the Early Stages of Life and the
Effects on Health
Later in Life
Christopher Stewart, Newcastle University, Newcastle upon Tyne, United Kingdom
Following birth, the infant gut is rapidly colonized by a range of microbes that
play fundamental
roles in health and disease. Unlike infants born at term (>37 weeks gestation),
extremely
preterm infants (<32 weeks gestation) have immature intestinal architecture and
an underdeveloped immune system. Because the preterm gut can become leaky,
translocation
of microbes into the bloodstream and/or intestinal cell death represent major
problems in this
29 | Page
Presentation Abstracts
vulnerable population. However, certain types of bacteria may promote gut and
immune
maturation.
Our group utilizes systems biology, combining microbiology, molecular biology, and
biochemistry to comprehensively profile clinical samples, including maternal breast
milk and
infant respiratory and gut samples. Building on from these association-based
analyses, we are
developing a state-of-the-art co-culture technology, allowing human intestinal stem
cell derived enteroids (�mini guts�) to be tested under physiologically relevant
oxygen
conditions. Using this ex vivo model will allow for targeted experimentation,
exploring how
specific microbes and milk components modulate intestinal barrier integrity and
function. Understanding how bacteria and gut epithelial cells interact holds
exciting
possibilities to better predict, diagnose, and treat infants at risk of disease.
2:15 p.m. Invited Speaker: The Composition of the Perinatal Intestinal Microbiota
in Cattle
Mikael Niku, University of Helsinki, Helsinki, Finland
We investigated the early development of the bovine intestinal microbiota by
quantitative PCR
and sequencing of the bacterial 16S rRNA genes. Microbiological samples were
collected from
within the rectum using contamination-shielded swabs. Calves were first sampled
immediately
at birth. The sampling was repeated at 24 hours of age, when the calves were
separated from
the dams. Calves were then housed individually and consumed mixed milk until the
third
sampling at 7 days of age.
Recent studies have generated controversial results regarding the existence of a
prenatal
microbiota in mammals. To control the ubiquitous bacterial DNA contamination, we
quantified
and sequenced negative control swabs. Microbial sequences which were relatively
abundant in
the controls were deleted from all data as probable contaminants. Quantitative PCR
and
sequencing indicated that the calf intestine contains DNA from a diverse low-
abundance
microbiota already at the moment of birth, which probably represents the late fetal
status. The
newborn microbiota was largely composed of Firmicutes and Proteobacteria, with
smaller
contributions of Actinobacteria and Bacteroidetes. The newborn core microbiota
represented
known anaerobic gut microbes and typical initial gut colonizers or mucosa-
associated genera.
Members of Lactobacillales were also abundant.
The newborn 16S rRNA gene profile resembled dam oral rather than fecal or vaginal
vestibular
microbiota, although the sampling was performed before the cows were allowed to
lick the
calves. This has been observed in previous human and murine studies. We did not
compare the
newborns to the other parts of the cow proximal digestive tract. In a phylum-level
comparison
to a previously published study of the bovine digestive tract microbiota, our
newborns were
closest to the microbiota of the adult bovine small intestinal mucosa.
At 24 hours from birth, the 16S rRNA gene copy numbers were on average 7000 times
higher
than in the newborns. The intestine was rapidly colonized by Escherichia/Shigella
and Clostridia,
and the alpha diversity collapsed. The calf rectal microbiota was still closer to
the dam oral
microbiota than to dam fecal microbiota.
By 7 days, there was a further 14-fold increase in the 16S rRNA gene copy numbers.
The alpha
diversity was increasing but still much lower than in adult feces. The phylum-level
microbial
community structure already resembled adult fecal microbiota, composed almost
completely
of Firmicutes and Bacteroidetes. However, the bacterial genera were different,
including
Faecalibacterium, Bacteroides, Lactobacillus, Butyricicoccus and Bifidobacterium.
Faecalibacterium and Butyricicoccus were rare in the feces of our adult forage-fed
cows.
Lactobacillus and Bifidobacterium were much less abundant in the calves than in
human infants,
30 | Page
Presentation Abstracts
possibly reflecting the smaller concentration and different composition of
prebiotic
oligosaccharides in the bovine milk.
Our observations suggest that the calf intestine is seeded before birth with a
diverse
microbiota or at least microbial components, but the microbiota changes rapidly in
the early
postnatal life. The early microbiota may serve to activate the developmental of the
immune
system and/or induce tolerance for commensal antigens. We are currently carrying
out
microbial culture experiments to investigate the presence of live bacteria in
newborn and
fetal calf samples.
3:15 p.m. The Composition of the Postprandial Metabolome of Human Blood and Urine
Depends
on the Human Genome and the Dairy Products Consumed
Guy Verg�res, Agroscope, Bern, Switzerland
Measuring the metabolome of bio-samples taken from people having ingested food
provides
information on both food composition and the individual metabolism of the consumer.
This
analytical strategy is particularly interesting for dairy research in order to
characterize the
biochemical processes taking place sequentially, firstly during the technological
transformation
of milk to dairy products, secondly during the gastrointestinal processing of the
ingested
products. In particular, as both processes involve transformation of the food
ingredients by
microbial consortia, combining metabolomics with microbial genomics allows to
investigate the
functional relationship between the food- and gut-associated microorganisms.
We have taken this approach in two human intervention studies investigating the
postprandial
serum and urine metabolome, measured by GC-MS and LC-MS, of healthy subjects having
ingested milk, yoghurt, and cheese. A significant fraction of the serum metabolome
changes
upon food ingestion, most of these changes being independent of the ingested dairy
product.
Processing of milk to yoghurt and cheese results, however, in a differential
postprandial
response for hundreds of molecules, the identity of most of them remaining unknown.
Among
the metabolites identified, fermentation of milk increases postprandial amino acids
and
dipeptides, indicating that this treatment can be considered as an ex vivo pre-
digestion. This
conclusion is supported by the size distribution of the molecules appearing in
blood as smaller
molecules are present at higher concentrations after yoghurt and cheese ingestion
compared
to milk. Transformation of milk to yoghurt also modulates the postprandial
concentration of
several bile salts indicating that milk fermentation interacts with the metabolic
activity of the
gut microbiota on these endogenously produced regulators of lipid metabolism. Such
an
interaction is also apparent with postprandial molecules derived from the microbial
metabolism
of milk-derived nutrients, including immune-modulating indoles, derived from
tryptophan, and
the putative cardiovascular risk factor trimethylamine N-oxide5, derived from
choline/betaine.
Among the metabolites responding to the intake of milk and yoghurt, carbohydrates
are
particularly interesting as their postprandial response not only depends on milk
processing but
also on the genetic background of the subjects. In particular, the serum and urine
concentrations of galactitol and galactonate, two liver metabolites of galactose,
reflect on the
ability of the subjects to digest lactose. Due to their phenotypic nature, these
tests are more
relevant than genetic tests to detect lactose maldigestion.
The postprandial serum and urine metabolome of the subjects also reveal the
intriguing
presence of fucosylated di- (blood group H disaccharide) and tri- (Lewis a antigen)
saccharides
derived from milk oligosaccharides. These molecules are present in human milk and
modulate
the development of the gut environment in the infant. However, their presence had
so far not
been reported in human adults after ingestion of bovine milk. Interestingly, we
observed a
striking dichotomic inter-individual variability in the appearance of these two
molecules in the
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Presentation Abstracts
blood and urine of the subjects. As milk-based oligosaccharides are primarily
digested by the
gut microbiota, the inter-individual postprandial variability in blood group H
disaccharide and
Lewis a antigen measured in blood and urine provides support for a role of genetic
polymorphisms of intestinal fucosyltansferases in modulating the capacity of the
gut
microbiota to digest milk-derived oligosaccharides. Given that these polymorphisms
have been
associated with a range of viral and bacterial gastrointestinal infections, the
ability of human
subjects to produce, upon bovine milk ingestion, putative decoy molecules for these
pathogens
raises interesting possibilities towards personalized dairy nutrition.
3:45 p.m. Student Travel Award Recipient: Infant Commensal Complexed with Milk
Secretory
Immunoglobulin A Prevents Enteric Infection
Vanessa Dunne-Castagna, University of California-Davis, CA, USA
Enteric infections plague millions worldwide each year, with the highest rates of
morbidity and
mortality in infants under 1 year of age. Breast milk provides marked protection
against enteric
pathogens through several mechanisms including antimicrobial proteins like
lactoferrin and
lysozyme, oligosaccharides to promote colonization of commensals that competitively
exclude
pathogens, and secretory immunoglobulin A (SIgA) known to supply passive immunity
to the
neonate. Recently, milk SIgA has gained new appreciation for its role in shaping
the commensal
biota through antigen-independent interactions, but to date there have been no
evaluations of
the impact of this relationship on enteric pathogens. This study aimed at
investigating the
functional outcome of a commensal:SIgA complex on the prevention of
enteropathogenic
invasion in human colonocyte cells in vitro, and in dampening the host inflammatory
response.
Invasion assays were completed by pre-treating a co-culture of Caco-2 colonocytes
and HT29-
MTX cells (90:10 ratio) with a commensal:SIgA complex confirmed through flow
cytometry
(1000 ug SIgA/1e7cfu ratio) for 1 h followed by inoculation with enterohemorrhagic
Escherichia
coli (EHEC), Salmonella enterica serovar Typhimurium (Salmonella), or Campylobacter
jejuni (C.
jejuni). A gentamycin protection assay eliminated extracellular bacteria for
Salmonella and C.
jejuni assays, and the colony forming units were counted from colonic cell lysates
to determine
invaded pathogen. Commensal pretreatment alone reduced Salmonella invasion by 50%
but had
no effect on EHEC adhesion and increased C. jejuni invasion. However,
commensal:SIgA
complex reduced Salmonella invasion by almost 90% (p<.001), EHEC adhesion by 45%
(p<.051)
and C. jejuni invasion by 100%. The commensal:SIgA complex reduced mammalian cell
gene
expression of the pro-inflammatory cytokine IL-8 by 6-fold and altered tight
junction binding
protein expression. In addition, the commensal:SIgA complex significantly reduced
the transepithelial electrical resistance (TEER), a measure of intestinal
permeability, when co-cultures
were pre-treated prior to Salmonella challenge. Mechanisms behind these effects
warrants
investigation. This may be through direct host-microbial crosstalk that strengthens
intestinal
barrier function, reduction of pathogen load, or alteration of metabolite secretion
of the
commensal and/or pathogen with SIgA. This study provides a better understanding of
the
multifarious roles of milk SIgA in shaping the infant gut microbial community and
may offer a
novel mechanism to prevent enteric infectious disease in the newborn.
4:05 p.m. Panel Discussion � Linking Sustainability to the Microbiome and
Metagenome
Moderator: Bruce German, University of California-Davis
Panelists: Troels Kristensen, Aarhus University; Nicole Darmon, INRA (tentative);
Monika Zurek,
Oxford University; Mikael Niku, University of Helsinki; Christopher Stewart,
Newcastle University
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Presentation Abstracts
4:30 p.m. Update from 2018�s Most Valuable Presentation: Human Milk
Oligosaccharides Directly
Enhance Gut Barrier Integrity to Alter Infections
Ishita Shah, University of California-Davis, CA, USA
Complex oligosaccharides present in human milk (HMOs), are the result of millions
of years of
mammalian evolution to protect the infant from severe infections. While HMOs have
many
functions, two main roles are as prebiotics for specific commensal bacteria and
reduction in
infection through serving as receptor-decoys for pathogens thus blocking their
binding to the
intestine and reducing rates of infection. However, we have made a recent discovery
that preincubation of specific HMOs like lacto-N-fucopentaose I (LNFP I) and 3�
sialyllactose (3� SL) with
intestinal cells directly enhances intestinal barrier integrity. We hypothesize
that the well
documented HMO-driven reduction in pathogen infection is modulated via two
mechanistic
routes, one via a direct interaction between HMO and intestinal cells thereby
improving barrier
function and the second through direct blocking of pathogen binding to the
intestine. Preincubation of LNFP I and 3� SL with intestinal cells directly
increases cell-cell adhesion, structural
integrity and dramatically reduces internalization of pathogenic bacteria.
Moreover, these
HMOs directly increase transepithelial electrical resistance indicating improved
tight-junction
dynamics and a barrier-strengthening effect prior to pathogen encounter. Direct
incubation
with these HMOs also results in an alteration in the infection-dependent signaling
cascade, and
a dramatic reduction in pathogen-mediated chemokine expression. Using a germ-free
mouse
model, we have shown that 3� SL supplementation of drinking water reduces Dextran
Sodium
Sulfate mediated intestinal injury as determined via specific alterations in
immune-markers in
the mouse cecum and colon. This new evidence suggests a previously unknown
protective role
for HMOs directly linked to intestinal health�a finding that will enhance
understanding of gut
health and maturation during not only breastfeeding but can be advanced to better
understand
a variety of gut diseases and therapeutics across age groups.
4:50 p.m. Highlights from IMGC�s �SPLASH!� milk science update� and Upcoming Hot
Topics
Danielle Lemay, USDA ARS Western Human Nutrition Research Center & University of
CaliforniaDavis
In April 2012, the IMGC began publishing an e-newsletter, �SPLASH!� milk science
update,�
which features four articles on emerging topics in milk science each month�that�s
48 new
articles on milk science each year. By the time of the IMGC conference in 2019, we
will have
published over 350 articles! This talk will reveal the editor�s choice awards
articles published in
the previous 12 months. It will also include a behind-the-scenes tour of SPLASH!:
who are the
current writers and editors, who are our readers and how do they reach our website.
The
SPLASH! newsletter has helped to grow the IMGC with more than 100,000 annual visits
to the
website each year. Nearly all traffic to the IMGC website is the result of SPLASH!
content. The
talk will also cover milk science topics expected to emerge in the coming year and
beyond.
Day 2 � Wednesday, November 13, 2019
8:30 a.m. Keynote Speaker: Composition, Structure and Digestive Dynamics of Milks
from
Different Species
Debashree Roy, Riddet Institute, Massey University, Palmerston North, New Zealand
Worldwide production of non-cow milks has nearly doubled in the last five decades,
substantiating the growing demand for these milks. The traditional dairy industry
is becoming
more and more diversified with non-cow milk and milk products, mainly because of
increasing
consumer perception about the health benefits of these milks. However, there is
very little
33 | Page
Presentation Abstracts
information available on the physicochemical and nutritional properties of the non-
cow milks.
We have studied the comparative composition, gel formation, and dynamic in vitro
and in
vivo digestion of cow, goat, sheep, red deer, and water buffalo milks available in
New Zealand.
We have found that the milks from these different species not only vary in their
composition of
macro and micronutrients, but also vary in their gelation profile, microstructures,
and digestion
behavior.
Sheep, deer, and buffalo milks had higher total solids and protein contents, which
resulted in
stronger gel formation on acidification compared with cow milk. In contrast, goat
milk
contained less protein and formed less firm gels on acidification compared with cow
milk.
Buffalo milk was an exception as, despite having lower protein and total solids
contents than
deer milk and sheep milk, it formed stronger gels than those from deer and sheep
milks.
The in vitro digestion study revealed that the milks from all these species formed
a firm
coagulum during gastric digestion and whole milks from all the species formed
softer curds
compared to the curds formed by skim milks. The amount of curd formed from
different milks
was a function of their total solids as well as protein (casein) content. As the
digestion
progressed and with the decrease in pH, the curd microstructure became more
cohesive and
the fat globules within the curd coalesced. Similar results were observed during
the in
vivo study.
Interestingly, the in vivo results also indicated that the rates of curd
strengthening for goat and
sheep milk were lower than cow milk in piglets� stomach. Besides, the
ultrastructure of the curd
formed from goat and sheep milk was more open compared to the curds formed by cow
milk
in piglets� stomach. These fundamental insights generated are a considerable step
forward in
understanding the nutritional properties of milks from different species. This
knowledge can be
used further for developing high-value dairy products for people of all age groups.
9:15 a.m. Invited Speaker: Functionality and Structural Aspects of Milk Genetic
Variants
Jared Raynes, Commonwealth Scientific and Industrial Research Organization (CSIRO),
Werribee,
Australia
The functionality of milk is determined by a complicated interplay of factors
including but not
limited to, stage of lactation, parity, season, type of feed and cow health.
Underlying these
factors are the genetics of the cow and in particular the genetic variations of the
milk proteins.
For example, it is well known that genetic variants of �- and ?-casein affect milk
coagulation.
How can variations of only 1 or 2 amino acids in the casein proteins, that are
intrinsically
unstructured, have such dramatic effects on milk functionality?
To elucidate how casein genetic variants alter milk functionality we have taken the
approach of
isolating individual casein genetic variants and investigate their properties with
a range of
advanced biophysical techniques. Research will be presented that provides evidence
of how
casein genetic variants elicit functional differences and potential opportunities
of how to
harness these differences for novel product development.
9:45 a.m. Genomic Variation in Milk Proteins in European Cattle Breeds
Bernt Guldbrandtsen, Aarhus University, Tjele, Denmark
The 1,000 Bull Genomes Project (Daetwyler, 2014) has created a global reference
dataset for
cattle breeds. The dataset in its sixth release contains whole genome variant calls
for more than
2,333 individuals belonging to Bos taurus. Animals have typically been sequenced to
a depth of
about 10X. Based on the literature, we identified a set of 39 genes on 21 autosomes
which either
34 | Page
Presentation Abstracts
code for milk proteins or for key enzymes affecting milk proteins and their
posttranslational
modifications such as glycosylases. We have annotated and filtered this dataset to
generate a
comprehensive map of coding-region variants, which are predicted to change or
possibly
change milk protein sequences. We examined 32 breeds, which were represented by at
least
five individuals. This included 2,044 animals, the rest representing crosses and
breeds with only
a few representatives.
In these 39 genes, 341 variants were observed. These had a quality of at least 50,
and they were
observed at least twice in the dataset. Among these were 220 simple (i.e.,
missense) variants
changing the amino acid sequence. Another 101 where splice region variants with
unpredictable, but potentially dramatic effects. Only one premature stop codon was
observed.
It was found in Fucosyl transferase 6 (FUT6) in BeefBooster and Gelbvieh. Four
frameshift
variants � also potentially highly damaging - were observed in xanthine oxidase,
lactoferrin,
tissue plasminogen activator, and perilipin 2. Missense variants were classified
using the SIFT
algorithm (Ng and Henikoff, 2003) which predicts the expected impact of a specific
amino acid
substitution. No less than 67 of the 220 variants were predicted to have large
effects on the
resulting protein (SIFT classification �deleterious�).
The genes with the highest number of protein changes (i.e., missense) were
butyrophilin
subfamily 1 member A1 (BTN1A1) and neuraminidase 2 (NEU2) with 16 variants while
perilipin 2
(PLIN2) had 15 variants. Among the major milk proteins, the numbers of variants
were aS1 casein
alpha (3), a S2-casein(3), �-casein beta (7), ?-casein (7), �-lactoglobulin (5), a-
lactalbumin (4),
and bovine serum albumin (5).
In whole dataset, 13,018 positions were observed as variable. In Holstein, 4,072
were
polymorphic (MAF at least 5%). Despite only eight individuals having been
sequenced, the breed
Machigiana had by far the highest number of DNA-level variants (5,909), whereas the
lowest
number was observed in traditional Danish Red (3,006). The latter breed is known to
be
relatively inbred. The second smallest number of DNA-level variants was observed in
Jersey
(3,340) and Angus (3,560) despite substantial numbers having been sequenced in each
of the
breeds.
Among the variants that change (or probably change) the protein product, each breed
was
polymorphic (MAF at least 5%) at between 39 (traditional Danish Red) and 82 sites
(Marchigiana). Holstein was polymorphic at 51 positions. Focusing on the missense
variants in
major milk proteins, the lowest number of polymorphic variants were found in
traditional
Danish Red and Deutsches Schwarzbuntes Niederungsrind (5), whereas the highest
number
was found in Marchigiana (14) and Chianina (14). Holstein had 7. Several beef
breeds were
polymorphic for a substantial number of variants (e.g., Belgian Blue 10, Charloais
12). Genome
sequencing identifies additional protein variants important for milk quality,
composition and
possibly functional properties.
10:45 a.m. Sequence-Based GWAS, Network and Pathway Analyses Reveal Genes Co-
Associated with
Milk Cheese-Making Properties and Milk Composition in Montb�liarde Cows
Marie-Pierre Sanchez, INRA, Jouy-en-Josas, France
Milk quality in dairy cattle is routinely assessed via analysis of mid-infrared
(MIR) spectra; this
approach can also be used to predict the milk�s cheese-making properties (CMPs) and
composition. When this method of high-throughput phenotyping is combined with
efficient
imputations of whole genome sequence data from cows genotyping, it provides a
unique and
powerful framework with which to carry out genomic analyses. The goal of this study
was to
use this approach to identify genes and gene networks associated with milk CMPs and
milk
composition in the Montb�liarde breed.
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Presentation Abstracts
Nine CMP traits (three measures of cheese yield, five coagulation traits, and milk
pH) and
contents of proteins, fatty acids, minerals, citrate, and lactose were predicted
from MIR spectra
in the From�MIR project. Phenotypes from primiparous Montb�liarde cows with at
least three
test-day records (1,506,037 test-day records from 194,934 cows) were adjusted for
non-genetic
effects and averaged per cow. 50K genotypes, available for a subset of 19,586 cows,
were
imputed in two steps: at the high density (HD) level using genotypes of 522
Montb�liard bulls
and then, at the sequence level using Run6 of the 1000 Bull Genomes Project
(comprising 2,333
animals including 54 Montb�liard bulls). The individual effects of more than 8.5
million variants,
imputed with high accuracy, were evaluated in a genome-wide association study
(GWAS) in a
mixed model including a random polygenic effect estimated using a genomic
relationship
matrix calculated from HD genotypes. The results of the GWAS were then subjected to
further
analysis via an Association Weight Matrix and the Partial Correlation and
Information Theory
(AWM-PCIT) approach. This strategy led to the detection of 59 QTL regions, most of
which had
highly significant effects on CMPs and milk composition. From the GWAS results, we
then
identified a set of 736 co-associated genes. Among these, the well-known caseins,
PAEP and
DGAT1, together with dozens of other genes such as SLC37A1, ALPL, MGST1, SEL1L3,
GPT, BRI3BP,
SCD, GPAT4, FASN, and ANKH, explained from 12�30% of the phenotypic variance of CMP
traits.
We were further able to identify metabolic pathways (e.g., phosphate and
phospholipid
metabolism and inorganic anion transport) and key regulators, such as PPARA, ASXL3,
and btamir-200c, that are functionally linked to milk composition. By utilizing an
approach that
integrated GWAS with network and pathway analyses at the whole-genome sequence
level, we
were able to highlight co-associated candidate genes that are functionally linked
to milk CMPs
and composition. Here, we propose candidate variants that explain a substantial
proportion of
the phenotypic variance of CMP traits and which therefore could be included in
genomic
evaluation models to improve milk CMPs in Montb�liarde cows.
This study was funded by the French Ministry of Agriculture, Agro-food and Forest
(CASDAR),
the French Dairy Interbranch Organization (CNIEL), the Regional Union of Protected
Designation cheeses of Franche-Comt� (URFAC) and the Regional Council of
BourgogneFranche-Comt�, under the project From�MIR.
11:15 p.m. Student Travel Award Recipient: Variability of Oligosaccharides in Milk
from Native
Nordic Cattle Breeds
Anne Vuholm Sunds, University of Aarhus, Tjele, Denmark
Milk oligosaccharides (OS) are complex sugars of high interest due to their
bioactive properties.
The OS play key roles in stimulating the growth of beneficial microorganisms in the
gut by their
prebiotic effects, contributing to brain development, as well as preventing
pathogen adhesion
in the gut. The content and complexity of OS are significantly lower in bovine milk
compared to
human milk, which leads to some challenges in the process of isolating bovine milk
OS for use
in e.g. infant formula.
Until now, OS profiling in bovine milk has been conducted on milk from modern,
high-yielding
dairy breeds, and no studies have profiled the OS distribution in milk from native
Nordic cattle
breeds. However, recent studies have shown that these breeds possess a great
phenotypic as
well as genotypic diversity that may also be reflected in their milk composition.
Compositional
variations have previously been shown for protein and fat. These Nordic indigenous
breeds
have evolved in different directions over thousands of years due to varying
climatic challenges
and their co-evolutionary history with humans. To address this knowledge gap, the
present
study investigated the OS content in milk from a unique sample set of eight native
Nordic cattle
breeds. This investigation included a qualitative characterization of OS. The milk
samples were
collected from native breeds originating from Norway (D�lafe cattle, Telemark
cattle), Sweden
36 | Page
Presentation Abstracts
(Mountain cattle), Denmark (Red Danish cattle), Iceland (Icelandic cattle),
Lithuania (native
Black- and White cattle) and Finland (Western Fincattle and Eastern Fincattle).
The analysis of OS was conducted by high-performance liquid-chromatography
chip/quadrupole time-of-flight mass-spectrometry (HPLC-Chip/Q-TOF MS). The study
showed a
significant variation (p = 0.05) in the amount of several OS between breeds. The
highest total
relative abundance of OS was seen in milk from the Western Fincattle, representing
approximately double the intensity that was detected in milk from conventional
cows. Further,
the results showed interesting variations in the OS distribution, sialylated and
neutral OS
showed the highest relative abundance in milk from the Western Fincattle, while the
highest
content of fucosylated OS was monitored in milk from Western Fincattle, Norwegian
D�lafe
cattle and Icelandic cattle. Milk from Red Danish cattle and Norwegian Telemark
cattle
contained higher relative abundance of sialylated OS compared to neutral OS,
whereas the
opposite was observed for the remaining breeds. In general, the breeds that are
known to be
closely related genetically tend to produce OS in a total relative abundance close
to each other.
Findings of the present study indicate that the high genetic divergence between
native Nordic
cattle breeds seems to be reflected in the resulting OS distribution.
12:30 p.m. Keynote Speaker: Casein Micelle Mineralization
Thom Huppertz, Friesland Campina, The Netherlands & Wageningen University, The
Netherlands
Nutritionally, the casein micelles in milk serve a dual purpose, as a supplier of
both essential
amino acids as well as essential elements, such as calcium and phosphate. The
presence of
centres of phosphorylated serine residues on as1-, as2- and �-casein allows them to
stabilize
nanoclusters of calcium phosphate. The nanoclusters form an integral part of the
casein
micelles and are responsible for linking primary casein particles to form the
substructure of the
casein micelles. For bulk milk, micellar calcium phosphate (MCP) has typically been
reported to
represent 5-6% of micelle mass a dry matter basis. However, when comparing the milk
from
individual cows, notable differences can be observed. The degree of casein micelle
mineralization was found to vary up to 2-fold between milk samples from individual
cows.
Milk samples with a lower degree of casein micelle mineralization contain more
casein in a nonsedimentable form, with no apparent specific enrichment for specific
caseins between
sedimentable and non-sedimentable casein. Correlations between casein micelle
mineralization
and casein micelle size were not observed. The degree of casein mineralization
impacts various
technological properties of milk, such as rennet coagulation and acid coagulation
properties.
For rennet coagulation, milk with a higher degree of casein mineralization shows
reduced
coagulation times and higher gel firmness for-induced coagulation and the firmness
and
microstructure of rennet and acid-induced gels formed from milk.
To allow identification of factors determining casein mineralization in milk,
predictive
equations were developed based on protein, calcium and potassium content of milk.
Using
this approach, various key factors determining casein mineralization were
determined,
including the breed of cows; mineralization was found to be higher in the milk of
Jersey than
Holstein-Friesian cows. Within both breeds, increasing protein content of the milk
and protein
yield from the cows were found to be correlated with a lower degree of
mineralization.
Furthermore, significantly higher mineralization was found for milk samples with a
higher
degree of phosphorylation of as1-casein (9P vs. 8P) and as2-casein (12P vs. 10P and
11P). Genetic
variants of the caseins were not found to affect the degree of casein
mineralization. For milk
from Dutch Holstein-Friesian cows, almost 50% of variation could be explained based
on
genetic factors.
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Presentation Abstracts
Variation in casein micelle mineralization appears to be a previously poorly
acknowledged
factor contributing to differences in technological properties between milk samples
and
warrants further study. The large percentage of variation explained by genetic
factors opens
opportunities for breeding strategies towards desired levels of mineralization.
1:15 p.m. Invited Speaker: Milk Cathepsins � Implications for Health and
Functionality
Lotte Bach Larsen, Aarhus University, Tjele, Denmark
Bovine milk contains a range of intrinsic or indigenous (i.e. bovine) proteolytic
enzymes and
enzyme systems. Apart from plasminogen system, bovine milk also contains
cathepsins,
secreted from the somatic cells, leaked from dead cells or even secreted by mammary
epithelial
cells. Of these the most well described is cathepsin D. The cathepsins belong to
various classes,
and exhibit different specificities and requirements for activity, as well as
segregate to different
fractions during processing. Thereby they can affect the yield, quality and shelf
life of milk and
dairy. On the other hand, biologically speaking, they may have physiological roles
linked to predigestion in the infant, formation of bioactive peptides and
reorganization of mammary tissue.
The activation enigmas of the cathepsins is covered.
1:45 p.m. Student Travel Award Recipient: High Proportion of Transient Neonatal
Zinc Deficiency
(TNZD) Causing Alleles in the General Population
Yarden Golan, Technion-Israel Institute of Technology, Haifa, Israel
Loss of function (LoF) mutations in the zinc transporter SLC30A2/ZnT2 result in
impaired zinc
secretion into breast milk consequently causing transient neonatal zinc deficiency
(TNZD) in
exclusively breastfed infants. However, the frequency of TNZD causing alleles in
the general
population is yet unknown. Herein we investigated 115 missense SLC30A2/ZnT2
mutations from
the ExAC database, equally distributed in the entire coding region, harbored in 668
alleles in
60,706 healthy individuals of diverse ethnicity. To estimate the frequency of LoF
SLC30A2/ZnT2
mutations in the general population, we used bioinformatics tools to predict the
potential
impact of these mutations on ZnT2 functionality and corroborated these predictions
by a zinc
transport assay in human MCF-7 cells. We found 14 missense mutations that were
markedly
deleterious to zinc transport. Together with 2 conspicuous LoF mutations in the
ExAC database,
26 SLC30A2/ZnT2 alleles harbored deleterious mutations, suggesting that at least 1
in 2,334
newborn infants is at risk to develop TNZD. This high frequency of TNZD mutations
combined
with the World Health Organization-promoted increase in the rate of exclusive
breastfeeding,
highlights the importance of genetic screening for inactivating SLC30A2/ZnT2
mutations in the
general population for diagnosis and prevention of TNZD.
2:30 p.m. Invited Speaker: Understanding the Function of the Milk Fat Globule
Membrane
Carolyn M. Slupsky, University of California-Davis, CA, USA
The link between food and health is complex, particularly for the developing
neonate, as the
period after birth is the time when long term programming is occurring in the
neurologic,
immune and metabolic regulatory systems. Breastfeeding is known to have short- and
longterm benefits, and we have previously reported profound differences between
breast-fed and
formula-fed infants with respect to growth trajectory, immunological development,
succession of the gut microbiome and metabolism that indicate development of unique
metabolic phenotypes as a consequence of diet. To understand the impact of one
component
of milk, the milk fat globule membrane (MFGM), on the infant microbiome and
metabolism, we
analyzed samples from three clinical studies, two in which MFGM was supplemented to
infants
less than 6 months of age, and one in which MFGM was provided to infants between 6-
11
months of age. While MFGM had a minimal impact on the structure of the microbiome,
it did
impact its function, and ultimately positively impacted the metabolic phenotype of
infants
38 | Page
 Presentation Abstracts
consuming MFGM regardless of age. Our results suggest that MFGM is an important
component of milk and has an important role to play in shaping metabolism.
3:00 p.m. Isolation, Characterization, and Biological Properties of Milk
Extracellular Vesicles
Jan Trige Rasmussen, Aarhus University, Aarhus, Denmark
Assessments of the total amount of phospholipids (PL) in bovine milk vary from 0.9
to 2.3%
depending on the applied extraction method (R�se Gottlieb and Folch, respectively).
More than
50 years ago it was established that the membrane (MFGM) enveloping and
solubilizing milk fat
accounts for 60-65% of milk PL, whereas the remaining 35-40% resides in the skim
milk phase.
Even though existence of PL membrane structures in skim milk have been known for
this long
period, knowledge about their biological origin and possible functions in milk is
first now
emerging. Already in the beginning of the 1950�s Morton reported presence of a
specific type
of phospholipid particles in milk and named them �milk microsomes�. After that, the
origin of
�milk microsomes� were discussed, but it became generally accepted that skim milk
phospholipid components most probably originates from detached remnants of the
outer
layers of MFGs. Recent results contradict this notion, because milk from various
species have
been shown to contain nanosized PL structures in skimmed milk that are distinct
form MFGs,
and they are generically termed extracellular vesicles (EVs), because they are
likely to
encompass both exosomes and microvesicles (see Blans et al. 2017). Besides milk,
EVs have
been identified in a number of other biological fluids and rediscovered as an
important part of
intercellular communication, which involves genetic exchange of RNA and microRNAs.
Importantly, although MFGs and EVs are of different intracellular origin they are
both secreted
through the apical membrane in a way that result in overlapping content of
constituents. The
purpose of the present contribution is to provide an overview of data that we have
collected
and elaborate on their significance. We have lately established methods for gentle
and effective
isolation of milk EVs. Size exclusion chromatography proved effective in separating
phospholipid membrane vesicles from the primary casein and whey protein components
in two
differently obtained casein reduced milk fractions. Of which one is generated
without
implementation of ultracentrifugation. Milk EV isolates contain heterogeneous sized
vesicle
structures with membrane character, and are enriched in the EV markers lactadherin,
CD9, CD63
and HSC70. Lipid analysis shows that the EV isolates are different from the MFGM by
not
containing a triacylglycerol core and by presenting a diverging phospholipid
profile. Moreover,
the isolates are enriched in RNA with distinct from that of milk fat globules and
milk cells. The
EV isolates content of classical milk phospholipid membrane proteins was
investigated by
western blotting analysis and shows that MUC1, MUC15 and CD36 are more enriched in
bovine
milk EV isolates in comparison to the MFGM. Furthermore, lactadherin is outlined as
a general
milk PL-membrane marker, by its ability to bind strongly to phosphatidylserine.
To be able to address the biological properties we have established an assay to
estimate and
characterize extracellular uptake of milk EVs by cultured intestinal cells. Milk
EVs were
specifically labelled with fluorescent lactadherin. The uptake has been
characterized
extensively in relation to incubation time, dose, temperature, viability and cell
type. Intracellular
sorting has been studied microscopically by following the colocalisation of milk EV
with markers
for endosomal compartments and lysosomes.
3:30 p.m. Invited Speaker: The Role of Whey Protein in Postprandial Glycaemic
Control
Daniela Jakubowicz, Tel Aviv University, Israel
Background: Postprandial hyperglycemia and elevated daily glycemic excursions are
strongly
associated with HbA1c and with increased cardiovascular risk in type 2 diabetes.
Therefore,
39 | Page
Presentation Abstracts
along with weight loss, the diet for Type 2 Diabetes, should be focused on the
reduction of
postprandial glycemia and the hyperglycemic peaks across the day.
A high-calorie protein Breakfast, medium-sized lunch and small dinner is a proven
successful
strategy for Weight Loss, improved satiety and reduced overall postprandial
glycemia (PPG)
and HbA1c in obese and Type 2 Diabetes. Recently it has been suggested that in
addition to the
amount of protein, the quality and the source of the protein consumed in the
Breakfast has
importance for reduction of postprandial glycemia. Particularly whey protein exerts
potent
direct insulinotropic effect and through stimulation of glucagon-like peptide 1
(GLP-1) leading
to reduction of PPG in T2D.
Objective: To search whether in Type 2 Diabetes, a diet with high protein Breakfast
consisting
of Whey protein has a greater impact on Weight Loss, overall PPG, satiety and HbA1c
than
Breakfast with different protein sources.
Methods: 48 Type 2 Diabetes, 22 males, BMI 32.1 0.9 kg/m2
, aged 58.9 4.5 y, were randomized
to 3 isocaloric (1500 kcal) weight loss diets during 12 weeks. The 3 diets had
Breakfast (660 kcal),
Lunch (567 kcal) and Dinner (276 kcal) with the same composition at Lunch (% of
carb:prot:fat
20:45:25%) and Dinner (13:40:47%), but differed in the Breakfast composition and
protein
content. The 3 breakfast meals were:
1. CBd: High-carbohydrate Breakfast Diet, n=15, with 13 g protein in the Breakfast:
(% carb:prot:fat; 65:15:20%), i.e., ready-to-eat cereals.
2. PBd: High-protein Breakfast Diet, n=16, with 36 g protein in the Breakfast:
(% carb:prot:fat; 40:40:20%), i.e., eggs, tuna and cheese, and
3. WBd: Whey Breakfast Diet, n=17, with 36 g protein in the Breakfast
(% carb:prot:fat; 40:40:20%), i.e., whey protein shake.
Additionally, on 3 separate days between day 10 and 15 after diet initiation, all
participants
underwent 3 all day CBd, PBd and WBd meal tests, to assess PPG, insulin, intact
GLP-1 and satiety
after Breakfast, Lunch and Dinner.
Results: After 12 weeks, Weight Loss was -3.5�0.3 kg (-3.8%) in CBd, -6.1�0.3 kg (-
6.8 %) in PBd
and the greater, Weight Loss was found in WBd, -7.6�0.3 kg (-8.4%) (p<0.001).
Compared to
CBd, the % of Weight Loss in PBd was higher by 44%, and in WBd was greater by 55%
(p<0.001).
The reduction of HbA1c was lower in CBd by -0.36 � 0.04%, in PBd was -0.6 � 0.04%
and the
greater reduction was found in WBd, 0.89 � 0.05% (p<0.001). Compared to CBd the %
of the
reduction of HbA1c was greater by 41% in PBd, and by 64% in WBd (p<0.001). Compared
to CBd
the AUC for overall PPG was 12% lower in PBd and 19% lower in WBd (p<0.001). The
AUC for
overall Insulin was 27% higher in PBd and 38% higher in WBd (p<0.001), while the
AUC for overall
intact GLP-1 increased more than in CBd, by 32% in PBd and by 41% in WBd (p<0.001),
similarly
the AUC for overall satiety was enhanced by 24% in PBd and by 30% in WBd compared
to CBd
(p<0.001).
Conclusions: This study demonstrates that increasing protein content from 13g to
36g at
Breakfast has a significant impact on Weight Loss, overall satiety, HbA1c and on
overall PPG in
Type 2 Diabetes. However, for the same protein content, Whey protein vs other
protein
sources, yields additional benefits on Weight Loss, overall satiety, reduction of
PPG and HbA1c.
Whey protein should be considered an important adjuvant in the management of Type 2
Diabetes.
40 | Page
Presentation Abstracts
4:00 p.m. Natural Glycosidases in Bovine Milk
S�ren Drud-Heydary Nielsen, Aarhus University, Tjele, Denmark
Besides lactose, sugars are present in bovine milk as free oligosaccharides or
glycoconjugates
bound to either proteins or lipids. These sugars have been associated with
antiviral,
immunomodulatory and prebiotic properties. The complex sugars are therefore
interesting and
of potential economic value for the industry, especially in the production of
infant formula.
Glycosidases are naturally present in bovine milk. These enzymes hydrolyses the
glycosidic
bonds in free oligosaccharides and glycoconjugates, and their presence in milk is
mainly
believed to be derived of bovine origin but may also be microbial. Knowledge of
these
glycosidases and their origin are, apart from a few marker enzymes like N-acetyl-
�glucosaminidase (NAGase) (Hovinen et al. 2016), rather limited. Since the
glycosidases cleaves
and modify sugar substrates in milk, they could affect their bioactive functions
and, therefore,
gaining more knowledge on the presence and activity of the various naturally
present
glycosidases in bovine milk would be highly valuable. Several genes coding for
bovine milk
glycosidases are known, and nine of these have previously been identified to have
significantly
increased gene expressions over lactation (Wickramasinghe et al. 2011). In milk,
the presence of
these enzymes has mainly been studied through enzyme activity assays, whereas
quantification
and exact identification of molecular forms and origins have been less documented.
Recent
advances in mass spectrometry techniques, makes it a technology that has the
capabilities for
detecting and quantifying these enzymes, even in the low abundances found in milk,
using
either untargeted proteomics or targeted proteomics by multiple reaction monitoring
(MRM).
In the present study milk samples were profiled for a range of glycosidase
activities in relation
to somatic cells and mastitis at quarter level. Milk was collected from both
healthy cows, as well
as from cows with either subclinical or clinical mastitis. We sampled from both a
reference
quarter and an affected quarter. Activity of eight different glycosidase enzymes
were analyzed
using fluorescence activity assays. The glycosidases with highest activities in
milk from healthy
cows were a-glucosidase, neuraminidase and NAGase. The glycosidases mostly affected
by
infection were NAGase and �-glucuronidase, as expected. Furthermore, significant
effects of
clinical mastitis were found for activities of fucosidase, �-galactosidase, a-
galactosidase and aglucosidase. Additionally, liquid chromatography mass
spectrometry (Orbitrap) were used to
analyze peptide profiles of tryptic digests of these milks. Specific peptides from
NAGase was
consistently identified in all milk samples and confirmed its presence in milk.
Additional
glycosidases of bovine origin were also identified, including �-glucuronidase, �-
mannosidase,
Hexosaminidase D, a-glucosidase, a-mannosidase and lysosomal acid
glucosylceramidase.
Interestingly, the activity of hexosamidase D has to our knowledge not previously
been
described in bovine milk and peptides from this protein were only identified in
milk from
infected quarters. Results from mass spectrometry analysis confirmed that a range
of
glycosidases deriving from bovine origin were present in the milk and we identified
their specific
protein ID. The identified peptides could, if suitable, be used for absolute
quantification of the
glycosidases. Mass spectrometry based on MRM is an approach well suited for this
purpose.
Day 3 � Thursday, November 14, 2019
9:00 a.m. Keynote Speaker: The Dairy Matrix and Health: Current Evidence and
Knowledge Gaps
Michelle McKinley, Queens University Belfast, Belfast, United Kingdom
Milk and dairy foods make a large contribution to population intakes of a wide
range of
nutrients. Each of the individual nutrients found in milk and dairy foods has an
important
function in the body. As well as considering the effects of isolated nutrients on
health, it is
important to evaluate the effects of whole foods in order to account for the effect
of the food
matrix. The concept of the food matrix encompasses the interactions between the
food
41 | Page
Presentation Abstracts
structure and the nutrients that sit within it. The nature of the food matrix will
influence
digestion, absorption and ultimately bioavailability of nutrients, thus the health
effects of food
may differ from what is predicted based on knowledge of its nutrient content.
In relation to milk and dairy foods, the concept of the dairy matrix proposes that
the unique
combination of nutrients and bioactive factors interact with each other and the
physical
structure they sit within to promote health, with the combined effects of the
matrix extending
beyond the sum of the individual parts. This presentation will review the evidence
for the dairy
matrix having a unique effect on health and will highlight evidence gaps.
9:45 a.m. Invited Speaker: Dairy Matrix Effects: Response to Consumption of Dairy
Fat Differs when
Eaten within the Cheese Matrix � a Randomized Controlled Trial
Emma Feeney, University College Dublin, Dublin, Ireland
Traditionally, nutrition research has examined the effect of individual nutrients
on aspects of
health. Although useful, this approach does not consider the interaction of the
nutrients within
a food, and with the overall food structure.
Evidence suggests that a �matrix� effect exists for many foods, whereby the
combination of
nutrients within a food leads to differential effects on health than when
considered in isolation.
This statement is particularly true for dairy foods, which contain unique
combination of
beneficial nutrients and bioactives in different structures with may interact to
confer a range of
different, positive, health benefits when consumed within the matrix of dairy.
Recent studies by our group in UCD have focused on cheese specifically, and the
effect on heart
health markers, and demonstrate that the matrix itself is vital for some of these
health benefits.
This talk will give an overview of our work to date and some of our planned future
work.
10:45 a.m. Discovering Added-Value Compounds in Dairy Streams: Proteomic Profiling
Reveals the
Presence of Xanthine Oxidase with Remarkable Antimicrobial Activity
Gulustan Ozturk, University of California-Davis, CA, USA
Over the past 15-20 years, prompted by consumers� desire for products higher in
whey proteins
and lower in fats and carbohydrates, the dairy ingredient industry has invested in
furthering the
purification of whey proteins and has achieved the production of whey protein at
improved
purity levels. Enhancements in membrane filtration processing technology have
enabled
production of whey protein concentrate virtually free of carbohydrate, saturated
fat, and
cholesterol. However, as a consequence of the additional processing steps, new
challenges are
raised by the emergence of resulting co-products, such as Procream, which are
poorly
characterized and have yet to be explored for their economic and nutritional value.
Procream
is a coproduct of whey protein isolate from the microfiltration of whey protein
concentrate and
is currently underutilized by the dairy industry. Procream represents 14 to 18% of
the total whey
processed by even a small manufacturer yielding approximately 32 million
tonnes/year. At the
same time, milk-derived antimicrobial compounds have gained both scientific and
commercial
interest as new bacterial and fungal diseases have sky-rocketed in most
industrialized countries.
Hence, there is a critical need for food-derived solutions to selectively eliminate
pathogens,
improve growth of desirable gut bacteria and reduce the overall inflammatory tone.
Unfortunately, the conventional medical approach is to exclude dairy products from
the diet of
affected individuals. In contrast, our team�s previous research has demonstrated
that milk
harbors a suite of bioactive compounds with the potential to modulate gut microbial
composition, promote healthy gut development, and reduce inflammation linked to
dysbiosis.
42 | Page
Presentation Abstracts
In average, Procream is composed of 50% protein (dry basis) and 12% fat � which is
possibly
derived from the milk fat globule membrane. Although the gross composition of the
Procream
has been studied, the characterization of specific proteins remains to be
elucidated. Therefore,
the objective of this work was to employ proteomics tools to unravel its protein
composition.
To our surprise, we identified 180 unique proteins: this represent the first
comprehensive
proteomic profiling of Procream in the literature. In particular, four proteins
derived from the
milk fat globule membrane were found in relatively high abundance: Butyrophilin,
Lactadherin,
Mucin-15 and the enzyme Xanthine dehydrogenase/oxidase.
Xanthine oxidase is considered to be the most abundant human milk protein component
of the
milk fat globule membrane and has been proposed to modulate the infants oral and
gut
microbiota. During nursing, the interaction between breast milk (source of Xanthine
oxidase)
and infant saliva (source of the substrates: xanthine and hypoxanthine) enables the
production
of nitrogen and oxygen reactive species with antimicrobial properties. This same
antimicrobial
mechanism via Xanthine oxidase was recently proven in bovine milk.
The identification of Xanthine oxidase as abundant protein in Procream was further
confirmed
by proteomic analysis of the trypsin digested SDS PAGE band corresponding to 140
kDa.
Intrigued by this finding, we assessed the activity of this enzyme by measuring the
rate of
oxidation of xanthine to uric acid in Procream and compared with bovine milk and
human
milk. Not only the activity of Xanthine oxidase in bovine milk turned out to be 20
times higher
than that of human milk, but in Procream, it was 60 times higher than that of
Xanthine
oxidase in human milk.
These findings indicated that Procream represents an opportunity for large-scale
recovery of
novel compounds for improving human health and purse innovations that go beyond the
traditional vision of dairy components used for nutritional purposes.
11:15 a.m. Student Travel Award Recipient: Maternal Allergy and the Presence of
Non-Human
Proteinaceous Material in Human Milk
Pieter Dekker, Wageningen University, Wageningen, The Netherlands
Human milk has previously been shown to contain proteins or protein fragments that
originate
from nonhuman organisms. This proteinaceous material could be a factor that is
involved in the
development of the immune system of the infant in the first stage of life, leading
to e.g.,
sensitization or the induction of allergen-specific tolerance. A relation between
maternal allergy
and breast milk composition could be involved in the development of allergic
diseases. This has
not yet been investigated with regard to nonhuman proteins or protein fragments.
The aim of
this study was to investigate the relation between maternal allergy and the levels
of nonhuman
proteins or protein fragments. For this, we reanalyzed data-dependent LC-MS/MS data
from
human milk serum protein samples of 10 mothers with atopic house dust mite allergy
and 10
non-atopic mothers. A search was carried out for peptide sequences originating from
bovine or
allergens, and label free quantification was used to test for differences between
the two
groups.
Out of the 80 nonhuman peptide sequences that were identified in this study, the
majority
(n=64) matched with Bos taurus proteins. The other 16 sequences matched with
proteins from
non-bovine allergens, but the majority of these identifications could not be
confirmed with
analysis of synthetized peptides. In the milk from allergic mothers, 10 peptide
sequences were
found in significant higher levels than in milk from non-allergic mothers. Nine of
these
sequences were annotated to bovine �-lactoglobulin.
43 | Page
Presentation Abstracts
The significant difference in nonhuman proteinaceous material between milk from
allergic and
non-allergic mothers shows that maternal allergy has an effect on the transfer of
this material
into the milk. This difference might have an effect on the development of the
immune system
of the suckling infant. Future studies need to be carried out to investigate the
human milk
protein composition and the relation with allergy development in the infant.
11:35 a.m. Panel Discussion � the Dairy Matrix
Moderator: Merete Myrup, Danish Agriculture & Food Council
Panelists: Michelle McKinley, Queens University Belfast; Emma Feeney, University
College
Dublin; Bruce German, University of California-Davis
44 | Page
Poster Abstracts
44 | Page
45 | Page
Poster Presentation Abstracts
1. Maternal Allergy and the Presence of Non-Human Proteinaceous Material in Human
Milk
Pieter Dekker, Wageningen University, Wageningen, The Netherlands
Student Travel Award Recipient
Pieter Dekker1
, Sjef Boeren2
, Jacques Vervoort2
, Kasper Hettinga1
1. Food Quality and Design Group, Wageningen University & Research, Wageningen, The
Netherlands
2. Laboratory of Biochemistry, Wageningen University & Research, Wageningen, The
Netherlands
Human milk has previously been shown to contain proteins or protein fragments that
originate from
nonhuman organisms. This proteinaceous material could be a factor that is involved
in the development
of the immune system of the infant in the first stage of life, leading to e.g.,
sensitization or the induction
of allergen-specific tolerance. A relation between maternal allergy and breast milk
composition could be
involved in the development of allergic diseases. This has not yet been
investigated with regard to
nonhuman proteins or protein fragments. The aim of this study was to investigate
the relation between
maternal allergy and the levels of nonhuman proteins or protein fragments. For
this, we reanalyzed datadependent LC-MS/MS data from human milk serum protein
samples of 10 mothers with atopic house
dust mite allergy and 10 non-atopic mothers. A search was carried out for peptide
sequences originating
from bovine or allergens, and label free quantification was used to test for
differences between the two
groups.
Out of the 80 nonhuman peptide sequences that were identified in this study, the
majority (n=64)
matched with Bos taurus proteins. The other 16 sequences matched with proteins from
non-bovine
allergens, but the majority of these identifications could not be confirmed with
analysis of synthetized
peptides. In the milk from allergic mothers, 10 peptide sequences were found in
significant higher levels
than in milk from non-allergic mothers. Nine of these sequences were annotated to
bovine �lactoglobulin.
The significant difference in nonhuman proteinaceous material between milk from
allergic and nonallergic mothers shows that maternal allergy has an effect on the
transfer of this material into the milk.
This difference might have an effect on the development of the immune system of the
suckling infant.
Future studies need to be carried out to investigate the human milk protein
composition and the relation
with allergy development in the infant.
2. Infant Commensal Complexed with Milk Secretory Immunoglobulin A Prevents Enteric
Infection
Vanessa Dunne-Castagna, University of California-Davis, CA, USA
Student Travel Award Recipient
Vanessa Dunne-Castagna1
, Jassim Al-Oboudi2
, David A. Mills2,3, Bo L�nnerdal1
1. Nutrition Department, UC Davis, Davis, CA
2 Department of Food Science & Technology, UC Davis, Davis, CA
3. Department of Viticulture and Enology, UC Davis, Davis, CA
Enteric infections plague millions worldwide each year, with the highest rates of
morbidity and mortality
in infants under 1 year of age. Breast milk provides marked protection against
enteric pathogens through
several mechanisms including antimicrobial proteins like lactoferrin and lysozyme,
oligosaccharides to
promote colonization of commensals that competitively exclude pathogens, and
secretory
immunoglobulin A (SIgA) known to supply passive immunity to the neonate. Recently,
milk SIgA has
gained new appreciation for its role in shaping the commensal biota through
antigen-independent
interactions, but to date there have been no evaluations of the impact of this
relationship on enteric
pathogens. This study aimed at investigating the functional outcome of a
commensal:SIgA complex on
the prevention of enteropathogenic invasion in human colonocyte cells in vitro, and
in dampening the
host inflammatory response. Invasion assays were completed by pre-treating a co-
culture of Caco-2
colonocytes and HT29-MTX cells (90:10 ratio) with a commensal:SIgA complex
confirmed through flow
46 | Page
Poster Presentation Abstracts
cytometry (1000 ug SIgA/1e7cfu ratio) for 1 h followed by inoculation with
enterohemorrhagic Escherichia
coli (EHEC), Salmonella enterica serovar Typhimurium (Salmonella), or Campylobacter
jejuni (C. jejuni). A
gentamycin protection assay eliminated extracellular bacteria for Salmonella and C.
jejuni assays, and the
colony forming units were counted from colonic cell lysates to determine invaded
pathogen. Commensal
pretreatment alone reduced Salmonella invasion by 50%, but had no effect on EHEC
adhesion and
increased C. jejuni invasion. However, commensal:SIgA complex reduced Salmonella
invasion by almost
90% (p<.001), EHEC adhesion by 45% (p<.051) and C. jejuni invasion by 100%. The
commensal:SIgA complex
reduced mammalian cell gene expression of the pro-inflammatory cytokine IL-8 by 6-
fold and altered
tight junction binding protein expression. In addition, the commensal:SIgA complex
significantly
reduced the trans-epithelial electrical resistance (TEER), a measure of intestinal
permeability, when cocultures were pre-treated prior to Salmonella challenge.
Mechanisms behind these effects warrants
investigation. This may be through direct host-microbial cross-talk that
strengthens intestinal barrier
function, reduction of pathogen load, or alteration of metabolite secretion of the
commensal and/or
pathogen with SIgA. This study provides a better understanding of the multifarious
roles of milk SIgA in
shaping the infant gut microbial community and may offer a novel mechanism to
prevent enteric
infectious disease in the newborn.
3. High Proportion of Transient Neonatal Zinc Deficiency (TNZD) Causing Alleles in
the General
Population
Yarden Golan, Technion-Israel Institute of Technology, Haifa, Israel
Student Travel Award Recipient
Yarden Golan1
, Adrian Lehvy1
, Guy Horev2
, and Yehuda G. Assaraf 1
1. The Fred Wyszkowski Cancer Research Laboratory, Dept. of Biology, and 2.
Bioinformatics Knowledge Unit
2. The Lorry I. Lokey Interdisciplinary, Center for Life Sciences and Engineering,
Technion-Israel Institute of
Technology, Haifa, Israel.
Loss of function (LoF) mutations in the zinc transporter SLC30A2/ZnT2 result in
impaired zinc secretion
into breast milk consequently causing transient neonatal zinc deficiency (TNZD) in
exclusively breastfed
infants. However, the frequency of TNZD causing alleles in the general population
is yet unknown. Herein
we investigated 115 missense SLC30A2/ZnT2 mutations from the ExAC database, equally
distributed in
the entire coding region, harbored in 668 alleles in 60,706 healthy individuals of
diverse ethnicity. To
estimate the frequency of LoF SLC30A2/ZnT2 mutations in the general population, we
used
bioinformatics tools to predict the potential impact of these mutations on ZnT2
functionality and
corroborated these predictions by a zinc transport assay in human MCF-7 cells. We
found 14 missense
mutations that were markedly deleterious to zinc transport. Together with 2
conspicuous LoF mutations
in the ExAC database, 26 SLC30A2/ZnT2 alleles harbored deleterious mutations,
suggesting that at least
1 in 2,334 newborn infants is at risk to develop TNZD. This high frequency of TNZD
mutations combined
with the World Health Organization-promoted increase in the rate of exclusive
breastfeeding, highlights
the importance of genetic screening for inactivating SLC30A2/ZnT2 mutations in the
general population
for diagnosis and prevention of TNZD.
4. Genetic Variants and Posttranslational Modifications of Milk Proteins in
Relation to Non-Coagulating
Milk
Kajsa Nilsson, Lund University, Lund, Sweden
Student Travel Award Recipient
K. Nilsson1
, L. Buhelt Johansen2
, S. I. Duchemin3
, M. Stenholdt Hansen2
, H. St�lhammar4
, W. F. Fikse5
, D. J. de Koning3
,
H. Lindmark-M�nsson1
, M. Paulsson1 and M. Glantz1
1. Department of Food Technology, Engineering and Nutrition, Lund University, Lund,
SE
2. Arla Foods amba, Aarhus, DK
3. Department of Animal Breeding and Genetics, Swedish University of Agricultural
Sciences, Uppsala, SE
4. Viking Genetics, Skara, SE
5. V�xa Sverige, Uppsala, SE
47 | Page
Poster Presentation Abstracts
The ability of milk to coagulate after addition of rennet is a crucial step in
cheese production. We have
previously shown that Swedish Red Dairy Cattle (RDC) has a high prevalence of
individuals, 18%, that
produce non-coagulating (NC) milk. This has a negative effect on cheese yield, and
thus it is of high
interest for dairies to eliminate NC milk. Previous studies have tried to link NC
milk to different genetic
variants of milk proteins and other studies have linked posttranslational
modifications (PTM) of milk
proteins to coagulation ability of milk. One type of PTM is glycosylation of ?-
casein, but the results from
earlier studies are contradictory, since it has been found that glycosylation of ?-
casein could both
increase or decrease the rennet-coagulation ability of milk. The aim of this study
was to determine the
protein profile in milk from Swedish RDC, including PTMs of the caseins, in order
to explore links
between the milk proteins and NC milk. Additionally, the results from this study
can be used to
investigate the overall protein profile of Swedish RDC cows and compare with other
breeds.
Milk samples from 619 individual Swedish RDC cows were analyzed using liquid
chromatography-high
resolution mass spectrometry and the obtained masses were compared to Uniprot
database to
determine genetic variants of both casein and whey proteins. PTMs were determined
by using masses,
where phosphorylations of the genetic variants of casein, as well as glycosylations
of ?-casein, were
obtained. Both the number of glycosylations as well as the glycosylation variant
(A-E) were determined.
The genetic variants and PTMs were combined with coagulation parameters measured in
the milk
samples, in order to distinguish the protein profile of NC milk from Swedish RDC.
In addition,
composition and physical properties of the milk samples were used in the study to
fully explore the
pattern for NC milk.
Genetic variants of milk proteins can be determined both genetically and
phenotypically, and by
measuring the variants in milk samples, the results will show which variants are
expressed at protein
level in the milk. Thus far, we have found that ?-casein variant E, which
previously has been shown to
have a negative impact on rennet-induced coagulation, has decreased in the Swedish
RDC breed. In our
data set, we have also found at least six different phosphorylation isoforms of
aS2-casein, which is very
uncommon. Further analyses should reveal additional insights into the relations
between detailed milk
protein profile and NC milk within Swedish RDC. By gaining more knowledge of
genetic variants and
PTMs of milk proteins affecting NC milk, this can be used in the breeding strategy
of Swedish RDC in
order to reduce the frequency of NC milk, and also to get a better understanding of
the mechanisms
behind the inability of milk to coagulate.
5. Variability of Oligosaccharides in Milk from Native Nordic Cattle Breeds
Anne Vuholm Sunds, University of Aarhus, Tjele, Denmark
Student Travel Award Recipient
Anne Vuholm Sunds1
, Apichaya Bunyatratchata2
, Randall Robinson2
, Maria Glantz3
, Marie Paulsson3
, Daiva
Leskauskaite4
, Anne Pihlanto5
, Ragnhild Inglingstad6
, Tove G. Devold6
, Gerd E. Vegarud6
, Bryndis Eva Birgisdottir7
,
Daniela Barile2
, Lotte Bach Larsen1
, Nina Aagaard Poulsen1
1. Department of Food Science, Faculty of Science and Technology, Aarhus
University, Denmark
2. Department of Food Science & Technology, University of California, Davis, USA
3. Department of Food Technology, Engineering and Nutrition, Lund University,
Sweden
4. Department of Food Science & Technology, Kaunas University of Technology,
Lithuania
5. Natural Resources Institute Finland
6. Norwegian University of Life Sciences, Faculty of Chemistry, Biotechnology and
Food Science, Norway
7. Faculty of Food Science and Nutrition, School of Health Sciences, University of
Iceland, Iceland
Milk oligosaccharides (OS) are complex sugars of high interest due to their
bioactive properties. The OS
play key roles in stimulating the growth of beneficial microorganisms in the gut by
their prebiotic effects,
contributing to brain development, as well as preventing pathogen adhesion in the
gut. The content and
complexity of OS are significantly lower in bovine milk compared to human milk,
which leads to some
challenges in the process of isolating bovine milk OS for use in e.g. infant
formula.
48 | Page
Poster Presentation Abstracts
Until now, OS profiling in bovine milk has been conducted on milk from modern,
high-yielding dairy
breeds, and no studies have profiled the OS distribution in milk from native Nordic
cattle breeds.
However, recent studies have shown that these breeds possess a great phenotypic as
well as genotypic
diversity that may also be reflected in their milk composition. Compositional
variations have previously
been shown for protein and fat. These Nordic indigenous breeds have evolved in
different directions
over thousands of years due to varying climatic challenges and their co-
evolutionary history with
humans. To address this knowledge gap, the present study investigated the OS
content in milk from a
unique sample set of eight native Nordic cattle breeds. This investigation included
a qualitative
characterization of OS. The milk samples were collected from native breeds
originating from Norway
(D�lafe cattle, Telemark cattle), Sweden (Mountain cattle), Denmark (Red Danish
cattle), Iceland
(Icelandic cattle), Lithuania (native Black- and White cattle) and Finland (Western
Fincattle and Eastern
Fincattle).
The analysis of OS was conducted by high-performance liquid-chromatography
chip/quadrupole time-offlight mass-spectrometry (HPLC-Chip/Q-TOF MS). The study
showed a significant variation (p = 0.05) in
the amount of several OS between breeds. The highest total relative abundance of OS
was seen in milk
from the Western Fincattle, representing approximately double the intensity that
was detected in milk
from conventional cows. Further, the results showed interesting variations in the
OS distribution,
sialylated and neutral OS showed the highest relative abundance in milk from the
Western Fincattle,
while the highest content of fucosylated OS was monitored in milk from Western
Fincattle, Norwegian
D�lafe cattle and Icelandic cattle. Milk from Red Danish cattle and Norwegian
Telemark cattle contained
higher relative abundance of sialylated OS compared to neutral OS, whereas the
opposite was observed
for the remaining breeds. In general, the breeds that are known to be closely
related genetically tend to
produce OS in a total relative abundance close to each other. Findings of the
present study indicate that
the high genetic divergence between native Nordic cattle breeds seems to be
reflected in the resulting
OS distribution.
6. Natural Variation in Milk Composition and Its Effect on UHT Storage Stability
Marije Akkerman, Aarhus University, Tjele, Denmark
Marije Akkerman1
, Nina A. Poulsen1
, Valentin M. Rauh2
, John S�rensen2 and Lotte B. Larsen1
1. Department of Food Science, Aarhus University, Blichers All� 20, P. O. Box 50,
DK-8830 Tjele, Denmark
2. Arla Foods Innovation Centre, Agro Food Park 19, DK-8200 Aarhus N, Denmark
Understanding and potentially exploiting the natural variations in milk composition
for the production
and differentiation into high value, clean label products of high quality and
stability outside the cool
chain is of great interest for the dairy industry. The quality and storage
stability of UHT milk is increasingly
being linked to the raw milk composition and quality. Several studies have
investigated UHT stability in
relation to major milk components. However, natural variation and the interplay
between proteins and
minor components in relation to UHT stability are less understood. The aim of the
present work was to
study how natural variations in citrate levels, calcium distribution, casein
micelle size and protein
compositions affect milk functionality and UHT storage stability.
Milk samples of individual cows from farms varying in feeding and management
practices were collected
from April to June 2017 to maximize natural variations in citrate and calcium. No
significant change in
citrate content was, however, observed during the sampling period, which suggests
that mechanisms
other than feeding affect citrate levels in milk. Several significant correlations
were found, including a
positive correlation between complexed serum calcium and total citrate, and a
negative correlation
between urea and ionic calcium.
Furthermore, based on the natural variation in citrate levels between individual
cows, milk samples
originating from cows with naturally high or low contents of citrate were subjected
to UHT treatment at
141�C for 4 s in a tubular heat exchanger, and stored at ambient temperatures for
up to 6 months.
49 | Page
Poster Presentation Abstracts
The overall milk composition, calcium distribution, pH and casein micelle size were
studied before and
after UHT treatment and during storage. After UHT treatment and during storage,
milks with naturally
high contents of citrate both had higher total serum calcium, as well as higher
levels of complexed serum
calcium, while the low citrate milks were characterized by larger casein micelle
size along with higher
conductivity.
Based on these results, pooled milk samples originating from cows milking either
large or small casein
micelles along with two randomly collected pooled milk samples were collected.
These samples were
subjected to UHT treatment and stored at 20�C and 40�C for up to six months. The
overall milk
composition, calcium distribution, pH, casein micelle size and protein profiles
were then studied before
and after UHT treatment in relation to ethanol stability and sedimentation. During
storage, the batch
with small casein micelles before the UHT treatment displayed significantly less
sedimentation
compared to batches with average and large casein micelles.
Protein profiles of the raw milk were further characterized by 2-DGE and LC-ESI/MS.
These results reveal
less ?-casein variants A and E in the batch with small casein micelles. This
suggest that the genetic
background of milk directly affects risk of sedimentation and thus UHT storage
stability.
7. Genetic Parameters for Rheological, Compositional and Physical Traits in Swedish
Red Dairy Cattle
Sandrine Duchemin, Swedish University of Agricultural Sciences, Uppsala, Sweden
S. I. Duchemin1
, K. Nilsson2
, H. St�lhammar3
, M. Stenholdt Hansen4
, H. Lindmark-M�nsson2
, W. F. Fikse5
, D-J de
Koning1
, M. Paulsson2 and M. Glantz2
1. Department of Animal Breeding and Genetics, Swedish University of Agricultural
Sciences, Uppsala, Sweden
2. Department of Food Technology, Engineering and Nutrition, Lund University, Lund,
Sweden
3. Viking Genetics, Skara, Sweden
4. Arla Foods amba, Aarhus, Denmark
5. V�xa Sverige, Uppsala, Sweden
The rennet-induced coagulation ability of milk is important for cheese production.
For Swedish Red Dairy
Cattle (RDC), this ability is reduced because of a high prevalence of non-
coagulating (NC) milk. The aim
of this study was to estimate genetic parameters for rheological, compositional and
physical traits as
well as for NC milk in Swedish RDC cows.
Blood and milk samples were collected during the stable period from 600 Swedish RDC
cows. For
genotyping purposes, DNA was extracted from blood samples and the cows were
genotyped for the
low-density chip (~10,000 SNP) and imputed to 50K SNP genotypes using a large
reference population
of genotyped animals (more than 50,000). From the milk samples, fat, protein,
casein, lactose, and citric
acid contents were analyzed using Fourier Transform Infrared spectroscopy and
somatic cell count (SCC)
was estimated using flow cytometry. Total calcium was estimated using inductively
coupled plasma
atomic emission spectroscopy, free Ca2+ using an ion selective electrode, buffering
capacity using
titration with NaOH, and casein micelle size using laser light scattering.
Furthermore, rennet-induced
coagulation was followed at 32�C where skim milk samples were pH adjusted to 6.5
prior addition of
chymosin. The rheological properties were measured using two different rheological
systems: a) free
oscillation rheometry (ReoRox4) was used to screen all milk samples, and b) low
amplitude rheometry
(Stresstech) was used for randomly selected samples. Parameters obtained from the
two systems (rox
and stress, respectively) were rennet coagulation time (RCT), gel strength (G�),
curd firming rate (CFR)
and yield stress (sy). Milk samples were defined as non-coagulating (NC) when
RCTrox > 40 min, and,
when occurring, NC milk was scored as a binary trait (NC=1; other=0). Variance and
covariance
components were estimated between NC milk and one of the other studied traits at a
time. The animal
model with genomic relationships was corrected for the fixed effects of parity,
weeks in milk, and
lactation curve, and with the random effects of herd and animal. A G-matrix built
from the 50K SNP
genotypes accounted for relationships between animals.
50 | Page
Poster Presentation Abstracts
The results show that bivariate heritabilities estimated of NC and the other
studied traits ranged from
0.25 to 0.31 (standard error (SE) 0.09 to 0.10). Furthermore, bivariate
heritabilities estimated for RCTrox
and the other studied traits ranged from 0.40 to 0.44 (SE 0.09 to 0.10).
Additionally, a total of 40
significant genetic correlations were found within the studied dataset. A
significantly strong positive
genetic correlation was found between NC milk and RCTstress (r=0.99), whereas
significant negative
correlations were found between NC milk and CFRrox, Gmax_rox, G�stress, sy_stress,
RCTstress and
total amount of calcium (r ranging from -0.98 to -0.65). Furthermore, calcium
significantly correlates
positively with CFRrox, Gmax_rox, G�stress, buffering capacity, casein content and
citric acid content (r
ranging from 0.49 to 0.89), and negatively with sy_stress and RCTrox (r ranging
between -0.47 and -
0.62). Lower amount of calcium can be linked to NC milk, thus showing reduced
coagulation ability
of milk. These results indicate that more detailed traits on milk composition as
well as physical and
rheological properties could be used within the breeding work to reduce the
frequency of NC milk.
8. Quantifying Breast to Breast Milk Volume in Effective Milk Transfer During
Breastfeeding Utilizing
New Bio Sensor Technology
Jimi Francis, University of Texas at Tyler, USA
Dr. Jimi Francis1
, Dr. Ruth Lucas2
, Dr. Arturo Arce-Esquivel1
, Dr. Premananda Indic1
, Dr. Prabha Sundaravadivel1
1. University of Texas at Tyler, USA
2. University of Connecticut, USA
Original Research - This presentation will provide information on maternal
potential failure points in
relation to human milk transfer in real time. Evaluation of the mammary tissue to
determine milk velocity
and flow was assessed using newly developed biosensors during breastfeeding.
The breastfeeding mother/infant dyad is a biologic system of interdependency. As a
living system, milk
transfer during sustained breastfeeding requires maternal effectiveness (production
and movement of
milk out of the breast) and infant efficiency (volume of milk swallowed over time)
to ensure infant
growth and health. Malfunctions during milk transfer may occur on either the
maternal side, the infant
side, or both, leading to early weaning. Breastfeeding is pivotal to optimal infant
health in the newborn
period and throughout the first year of life. Despite high rates of breastfeeding
initiation after birth,
sustained breastfeeding rapidly decreases overtime and by two weeks 30% of the
mothers have begun
adding artificial milks to infant feedings. Therefore, mothers and healthcare
providers have a pressing
need for objective methods to assess milk transfer during breastfeeding. To address
the maternal side
of the equation, evaluation of milk flow and velocity from the human breast was
assessed using newly
developed non-invasive procedures and techniques to evaluate potential points of
failure during
breastfeeding.
This study collected data on healthy full-term infants breastfeeding well to
evaluate the normative
ranges of milk velocity and flow during breastfeeding. The methods included the use
of ultrasound and
mass of milk transfer to validate the use of biosensors to measure milk velocity
and flow from the breast.
This project is part of developing a breastfeeding diagnostic device that captures
the effectiveness of
milk ejection through the delivery of the milk bolus to the infant's stomach with
the goal of evaluating
neurologic development in the neonatal period based on an infant's feeding
organizational skills. The
use of biosensors is a developing area in medical technology, and this is a novel
way to assess
breastfeeding at points of intervention to improve breastfeeding outcomes.
51 | Page
Poster Presentation Abstracts
9. Alpha-lactalbumin Enriched Whey Protein Concentrate to Improve Gut, Immunity and
Brain
Development in Preterm Pigs
Charlotte Holme Nielsen, University of Copenhagen, Copenhagen, Denmark
Charlotte Holme Nielsen1
, Duc Ninh Nguyen1
, Anne B. Lau Heckmann2
, Per Torp Sangild1
, Thomas Thymann1
, Stine
Brandt Bering1
1. Comparative Pediatrics and Nutrition, University of Copenhagen, Denmark
2. Arla Foods Ingredients, Viby, Denmark
Background: Preterm birth predisposes to developmental complications including
immature
gastrointestinal and immune functions, postnatal growth restriction and delayed
neurodevelopment.
Human milk is important for neonatal development, but infant formula
supplementation may be needed
to secure proper growth. Alpha-lactalbumin (a-La) is a major component of human
milk (~4 g/L in early
milk), and its biological activity may contribute to the benefits of breastfeeding.
We hypothesized that
supplementation of milk with an a-La-enriched whey protein concentrate (WPC) would
stimulate gut,
immune and brain development in preterm neonates.
Methods: We tested this by feeding cesarean-delivered preterm pigs (90% gestation)
dilute bovine milk
(2:1 in water) without (REF group, n = 22 from two separate litters), or with a-La
enrichment of WPC
provided at two different levels (HIGH 18 g/L, LOW 6.3 g/L, n = 19-20). Total
protein contents was 27 g/L
(REF) and 55 g/L in the two enriched diets. All pigs were reared by identical
procedures, and clinical
variables and functional endpoints (e.g., T-maze cognition test) were assessed at
intervals during the
study. Gut microbiota and organ weights were recorded at day 19.
Results: Both HIGH and LOW pigs grew faster than the REF pigs, but with no
difference between the
two supplemented groups (29 vs. 19 g/kg/d). The HIGH pigs had higher bone mineral
composition and
density but lower relative intestinal weights and proximal villus heights than LOW
pigs. Other gut
endpoints (e.g., intestinal brush border enzymes, goblet cell density) and immune
parameters (e.g.,
phagocytic capacity, lymphocyte subsets) were similar between REF, HIGH and LOW
pigs. Likewise,
cognition outcomes (open field, spatial T-maze tests) did not differ between HIGH
and LOW groups, and
both were similar to values in REF pigs. Relative to LOW pigs, the HIGH pigs had
higher colon microbiota
alpha-diversity and levels of acetic and butanoic acid and tended to have reduced
gut transit time (p =
0.07).
Conclusion: Protein supplementation of milk with WPC improved growth in preterm
pigs. Further
enrichment with a-La, beyond the levels in human milk, had marginal effects on food
transit and gut
microbiota, but did not affect gut, immunity or brain functions. WPC may be a good
supplementary milk
protein source for both preterm and term infants.
10. The Effect of Different Time-Temperature Profiles on Bioactive Proteins During
Pasteurization of
Donor Human Milk
Eva Kontopodi, Amsterdam UMC, University of Amsterdam, Emma Children�s Hospital,
Amsterdam, The
Netherlands
Eva Kontopodi1
, Kasper Hettinga2
, Bernd Stahl3
, Johannes B. van Goudoever1,4, Ruurd Van Elburg1,3
1. Amsterdam UMC, University of Amsterdam, Emma Children�s Hospital, Amsterdam, The
Netherlands
2. Dairy Science and Technology Group, Wageningen University, Wageningen, The
Netherlands
3. Danone Nutricia Research, Utrecht, the Netherlands
4. Amsterdam UMC, Vrije Universiteit, Emma Children�s Hospital, the Netherlands
Rationale: When mother�s own milk is unavailable, donor human milk represents the
best alternative
form of nutrition for preterm infants. To ensure safety, donor milk is generally
processed by holder
pasteurization (HoP) at 62.5�C for 30 min. This temperature-time combination may
negatively affect the
activity of some important bioactive milk components: immunological proteins are
only partially
52 | Page
Poster Presentation Abstracts
preserved and e.g. the enzyme bile salt stimulated lipase (BSSL) is completely
inactivated. In addition,
long warming up and cooling down times (lag-times) may also affect the bioactive
properties of
pasteurized milk. High-Temperature-Short-Time (HTST), a treatment with shorter
processing times (15
sec at 72�C), may be a suitable alternative to HoP. We aimed to compare the effects
of different heating
processes on bioactive proteins and the safety of human milk.
Methods: Five pasteurization methods were conducted: First, to compare the impact
of HoP and HTST
on the bioactive proteins and secondly, to study the influence of lag-time by
comparing pasteurization
methods with the same holding time as HTST (15 sec at 72�C), but with different
heating up and cooling
down times. To perform rapid pasteurization with shorter lag-times, a lab-scale
HTST pasteurizer was
built. The native protein concentration was measured with a BCA assay and further
identification was
obtained with LC-MS/MS. SDS-PAGE under reducing and non-reducing conditions was
performed and
the enzyme-activities of BSSL and alkaline phosphatase (ALP) were measured. To
investigate the effect
of the different pasteurization methods on the bacteriostatic properties, human
milk samples previously
treated with the five heating methods were inoculated with 2100 CFU/ml [Link] K12
and [Link].
Results: HTST generally performed better than HoP. The total native protein content
was found to
decrease at increasing lag-times, in all five methods (p<0.05). Lactoferrin and
BSSL were found to be the
most heat labile whereas lysozyme remained intact for the most part (p<0.05). The
SDS-PAGE gels
showed no protein aggregation in human milk serum. All pasteurized samples were ALP
negative, with
a significantly low BSSL activity (<10%). Raw milk had similar bacteriostatic
properties to HTST treated
samples, while for HoP samples, the multiplication rate of [Link] and [Link] was
double compared to
both HTST treated and untreated samples (p<0.05).
Discussion: After HTST, bioactive properties of human milk were better preserved
with similar safety
compared to HoP and the other pasteurization methods. The difference in lag-time
(warming up and
cooling down) was more important in the preservation of bioactive proteins than the
difference of the
heating processing itself (time-temperature combinations) among the investigated
methods. In
conclusion, a treatment with considerably short lag-times, such as HTST, may reduce
the thermal
damage caused to the bioactive proteins compared to HoP, without affecting safety.
11. Colostrum for Gut Protection Before or Repair After Formula Feeding in Preterm
Neonates
Yanqi Li, University of Copenhagen, Copenhagen, Denmark
Presented by Stine Brandt Bering
Yanqi Li1
, Xiaoyu Pan1
, Duc Ninh Nguyen1
, Shuqiang Ren1
, Per Torp Sangild1,2,3
1. Comparative Pediatrics and Nutrition, University of Copenhagen, Copenhagen,
Denmark
2. Department of Pediatrics and Adolescent Medicine, Rigshospitalet, Copenhagen,
Denmark
3. Hans Christian Andersen Children�s Hospital, Odense University Hospital and
University of Southern Denmark
Background - Mother�s own milk (MM) improves gut health and protect against
necrotizing enterocolitis
(NEC) in preterm infants. Due to absence or insufficient supply of mother�s milk,
many infants often
receive sub-optimal formula (FOR) products before or after MM feeding during the
first weeks. FOR
consists of many highly processed ingredients, e.g. bovine milk protein combined
with vegetable oils
and carbohydrates, potentially leading to loss of important milk bioactivity and
synergetic effects of
components found in intact milk. Intact bovine colostrum (COL) has been shown to
improve gut health
and maturation in preterm pigs. Using preterm pigs as model for sensitive infants,
and COL to reflect the
high bioactivity of MM, we tested if COL feeding before or after FOR feeding
protects against FORinduced gut damage.
53 | Page
Poster Presentation Abstracts
Methods - Seventy-four preterm pigs received increasing volumes of COL or FOR until
d 5. At this age,
pigs were euthanized or fed either COL or FOR for another 4 d, resulting in six
groups: COL or FOR until
d 5 (C5, F5, both n=11), COL or FOR until d 9 d (CC, FF n=12-13), COL followed by
FOR (CF, n=14) and FOR
followed by COL (FC, n=13). Blood and tissues were collected on d 5 and d 9 for
measurement of clinical
variables together with structural, functional and gene expression parameters in
the intestine.
Results - FOR reduced the weight gain and physical activity, and increased
necrotizing enterocolitis (NEC)
incidence on d 5, relative to COL (64 in F5 vs. 27% in C5, P<0.05). Intestinal
structure and function (e.g.
villus height, crypt depth, brush border enzymes, hexose absorption, intestinal
integrity) were reduced
in F5 vs. C5 pigs, together with evidence of a proinflammatory response (increased
innate immune gene
expressions, such as LBP, MYD88, IL8, C3). On d 9, the incidence of NEC were
similarly low in all groups
(15-21%), but the CC, FC, and CF pigs showed improved intestinal structure and
function, relative to FF
pigs (P<0.05). Gene expressions levels related to tissue hypoxia (HIF1A) and
apoptosis (CASPS) were
similar between groups on d 5. On d 9, HIF1A expression was increased in FC and FF
groups, relative to
CC and CF, and CASP3 expression was increased in CF and FF vs. CC and FC pigs
(P<0.05).
Conclusion - Intact milk diets with high bioactivity, like COL and MM, may protect
the immature intestine
by dampening innate inflammatory pathways. Diet-dependent inflammatory sensitivity
is highest within
the first week after preterm birth and may decrease thereafter. Early, moderate gut
inflammatory
lesions may be reversible. Feeding early COL or MM may improve intestinal structure
and function for a
later FOR-induced insult (hypoxia, apoptosis). Conversely, COL or MM may help to
prevent the damage
caused by feeding FOR just after birth.
12. Oxytocin Receptor Gene Variants and Breast and Nipple Pain During Breastfeeding
Ruth Lucas, University of Connecticut School of Nursing, Storrs, Connecticut, USA
Ruth Lucas1
, Yiming Zhang1
, Stephen Walsh1
, Erin Young1
, Angela Starkweather1
1. University of Connecticut School of Nursing
Purpose: Single nucleotide polymorphisms (SNPs) within catechol-O-methyltransferase
(COMT) and
oxytocin receptor genes (OXTR) are associated with pain sensitivity in various pain
conditions. The
purpose of this study was to describe the relationship between COMT SNPs (rs6269,
rs4633, rs4818 and
rs4680), OXTR SNPs (rs2254298 and rs53576) and nipple and breast pain by women
during breastfeeding
within 48 hours, 1, 2, and 6 weeks after delivery.
Design: A descriptive design was used.
Sample: 65 women were enrolled in our Breastfeeding Self-Management (BSM)
intervention study for
managing nipple and breast pain during breastfeeding at 1, 2, and 6 weeks after
discharge.
Methods: Within 48 hours of delivery, women completed the Brief Pain Inventory
(BPI), a visual analog
scale (VAS) of nipple and breast pain severity, quantitative sensory testing (QST),
and buccal swabs were
collected from all patients. The BSM intervention began after discharge using a
cloud-based platform to
deliver knowledge, practice skills and provide support through nurse lead text-
based communication to
decrease pain intensity and increase adaptive coping behaviors.
Quantitative Sensory Testing (QST), a non-invasive, pain-free technique that is the
gold standard in
research to assess chronic pain and can assist in early detection of peripheral
sensory disorders, was
used to assess mechanical and pressure pain sensitivity. The QST protocol used
seven touch and pressure
tests to identify women at high risk of pain-associated breastfeeding cessation.
54 | Page
Poster Presentation Abstracts
Results: Significant associations between OXTR SNPs at rs2254298 (G/G = 30, A/G=
27, A/A = 3) and VAS
ratings (????12= 4.39, P = 0.036) was found using Likelihood Ratio Test based on
Linear Mixed Model with
demographic adjustment. Positive effect of number of minor allele of rs53576 on VAS
rating is also
detected (Estimate = 6.14, SE = 3.14). A negative significant correlation based on
spearman correlation
between QST subscale ALL (a painful sensation caused by innocuous stimuli) and
rs53576 was found (r
= -0.30, P = 0.021). There was no significant difference in breastfeeding duration
across time. The COMT
genotype was not associated with self-reported general pain.
Conclusions: Women carrying the OXTR minor allele rs53576 reported greater pain
sensitivity at baseline
and was predictive of increased self-reported breast and nipple pain during
breastfeeding across time.
Clinical Relevance: The present findings suggest that the previously identified
�risk alleles� for other pain
modalities (COMT genotype) may not have the same predictive relationship(s) with
breast and nipple
pain. This is the first study to consider oxytocin alleles as a factor contributing
to pain sensitivity which
increased the risk of breastfeeding cessation. Identifying risk factors, which
contribute to breast and
nipple pain, may allow for early-targeted interventions to reduce pain and promote
breastfeeding
success.
13. Infant Formula Subjected to Indirect Ultra-High Temperature (UHT) Treatment and
Long-Term
Storage May Adversely Affect Gut and Immune Development in Newborns
Jing Sun, University of Copenhagen, Copenhagen, Denmark
Jing Sun1
, Karoline Aasmul-Olsen1
, Duc Ninh Nguyen1
, Halise G�l Akillioglu2
, Marianne Nissen Lund2
, Christian Fiil
Nielsen3
, Mikael Christensen3
, Per Torp Sangild1
, Stine Brandt Bering1
1. Comparative Pediatrics and Nutrition, Department of Veterinary and Animal
Sciences, University of Copenhagen
2. Department of Food Science, University of Copenhagen
3. Arla Foods Ingredients, Denmark
Background: Ready-to-feed (RTF) liquid infant formula sterilized by UHT is
increasingly used to feed
preterm infants when maternal milk is not available. While UHT treatment
effectively eliminates bacterial
contamination, intensive thermal processing by indirect UHT-treatment and
subsequent long storage
period may reduce bioactivity and increase Maillard reaction products (MRP). Using
preterm pigs as a
model for milk-sensitive newborn infants, we investigated if an experimental
processing procedure,
using indirect UHT treatment coupled with relatively long storage at high
temperatures, would increase
MRP products in the formula and thereby negatively affect gut and immune
development just after
birth.
Methods: A model liquid formula subjected to low-temperature pasteurization (72-
75�C x 15 s, PAST),
indirect UHT sterilization (143 �C x 6 s, UHT), or indirect UHT sterilization
followed by storage in PET
bottles at 40�C for 60 days (S-UHT), was fed to cesarean-delivered preterm pigs
(90% gestation) for 5
days (24-120 mL/kg/d). Body and organ weights, blood leukocyte phenotyping, hexose
absorption,
digestive enzyme activities, severity of necrotizing enterocolitis (NEC) and
density of bacteria in bone
marrow (as a marker of systemic infection) were determined.
Results: The S-UHT formula had high content of all identified MRPs (i.e. furosine,
carboxymethyllysine,
carboxyethyl lysine, glyoxal, methylglyoxal derived hydroimidazolones) compared
with the other two
formulas (2.6-fold higher levels than PAST). Pigs had similar weight gain but S-UHT
pigs had more
diarrhea (p<0.05) and more NEC lesions (p<0.05 for distal small intestine), than
PAST pigs. The UHT-fed
pigs tended to have lower galactose absorption, higher gut permeability and more
bone marrow
bacteria, compared with PAST pigs (all p<0.15). Relative to PAST pigs, intestinal
lactase activity was
markedly reduced in S-UHT pigs (to 50%, p<0.05), followed by UHT pigs (p=0.06).
Less blood leukocytes,
neutrophils and monocytes were observed in S-UHT pigs (p<0.05), and a tendency to
more Treg cells
55 | Page
Poster Presentation Abstracts
(p=0.1),relative to other groups. Organ weights or clinical chemistry parameters
were minimally affected
by differences in formula treatment.
Conclusion: The indirect UHT treatment of liquid formula, coupled with long storage
at high
temperature, impairs gut and immune maturation in sensitive preterm pigs. This may
be mediated by
MRP accumulation in the model formula, in response to indirect UHT, coupled with
long storage at high
temperature. A milder processing condition (e.g. direct UHT treatment, shorter
duration and lower
temperature of storage), to reduce Millard reactions, may improve physiological
responses in milksensitive newborn pigs and infants.
14. Analytical Techniques to Study the Milk Metabolome and Lipidome for Insights
into Milk Bioactives
Ulrik Sundekilde, Aarhus University, Aarslev, Denmark
Background and hypothesis: The birth of a living human being is the result of an
approximately ninemonth pregnancy in which the developing foetus has taken exactly
the building blocks necessary to
grow and develop from its mother. However, growth and development continues in
multiple dimensions
at an increasing pace after birth. The nutrition in the first 1,000 days from
conception to the child�s 2nd
birthday plays a pivotal role in shaping the future health of the child. Yet, we
know little of how breast
milk components vary due to maternal factors or of the biological mechanisms behind
the beneficial
actions of many breast milk nutrients. For more detailed knowledge of milk
bioactives there is a need
for modern analytical techniques; one of which is metabolomics. Metabolomics is the
analysis of lowmolecular weight compounds, of which many are bioactives.
Methods: In this talk, data from a study on human milk collection study from the
mothers of pre-term
infants will be presented. The milk samples were analyzed by 1H-NMR spectroscopy1
to characterize the
polar metabolites and the phospholipids were analyzed by targeted lipodomics using
LC-MS2.
Preliminary metabolomics data from a new mother-infant cohort will also be
presented.
Results: Metabolite profiling revealed differences between pre-term and full-term
milk in the levels of
specific metabolites including carnitine, oligosaccharides, citrate, choline and
formate. Both gestational
age and age postpartum affected the phospholipid (PL) composition of human milk.
Significantly higher
concentrations (p < 0.05) of phosphatidylcholine (PC), sphingomyelin (SM) and total
PL were found in
preterm milk throughout lactation, as well as significantly higher concentrations
(p < 0.002) of several
phosphatidylethanolamine (PE), PC and SM species.
Conclusion: The use of two complementary techniques in profiling milk for
metabolites and phospholipid
species enabled the identification of differences in milk compounds between mothers
delivering
prematurely compared with mothers delivering at term. Many of these metabolites and
PLs are known
bioactives, and their presence or absence may affect infant growth and development.
Presently, our focus is currently to explore larger cohorts to verify our results
in addition to examining
longitudinal milk samples from mothers of pre-term infants and term infants to
establish how the extent
of prematurity and early lactation influences the metabolome, microbiome and
lipidome of the milk.
Furthermore, we wish to investigate the significance of milk bioactives in shaping
the milk microbiome
and infant metabolism and gut microbiota.
56 | Page
Poster Presentation Abstracts
15. The Effect of Diet and Milk Processing on Dairy Milk Exosomes and Their
MicroRNA Cargo
Sirja Viitala, Natural Resources Institute Finland (Luke), Jokioinen, Finland
Santeri Kankaanp��1
, Daniel Fischer1
, Markus Nurmi1
, Oliver Bitz1
, Sari M�kinen1
, Sirja Viitala1
1. Natural Resources Institute Finland (Luke), Jokioinen, Finland
Mammary epithelial cells of all mammals produce and release nano- and microsized
membrane vesicles
to milk. Exosomes (EXs) are subgroup of extracellular vesicles that transfer
biomolecular cargo from
donor cells to recipient cells. EXs are secreted in every multicellular system via
specific and highly
conserved secretion pathway. Compelling evidence suggest that in milk EXs play
important role in
biological programming of the newborn by delivering various regulatory biomolecules
such as proteins
and RNA from mother to infant. Abundant milk miRNAs are highly conserved among
mammals and thus
it is feasible that milk consumers are exposed to inter-species regulation of gene
expression with an
unexpected impact. Many biological factors as well as milk processing may affect
the regulatory signals
milk EXs mediate. The aim of the study was to investigate the effects of cow�s diet
and milk processing
on dairy milk EX miRNA cargo. Ten Finnish Ayrshire cows were used in 5x3 Latin
square design with three
35d experimental periods. Experimental diets consisted of total mixed rations based
on timothy -
meadow fescue -grass silage containing either high (60/40) or low (40/60) forage to
concentrate ratio,
and red clover -grass silage containing high (60/40) forage to concentrate ratio.
Morning and evening
milk samples were collected on day 32 and 33 of each experimental period to study
the effect of diets.
To study the effect of milk processing tank milk was collected from Luke�s
experimental dairy farm and
processed in Luke�s pilot dairy plant. The EXs were isolated by differential
centrifugation and validated
using NTA and TEM along with western blot with specific markers. Further analysis
from vesicles was
done with Xevo G2 QTOF mass spectrometry to characterize the proteome of isolated
vesicles. Several
membrane proteins known to enrich to mammalian EXs were observed. Total RNA was
extracted from
isolated EXs and sequenced with Illumina HiSeq 2500 instrument using Truseq v2
Rapid sequencing
chemistry (1x75 bp). Several RNA biotypes, including various small and long non-
coding RNAs, rRNA,
tRNA and mRNA were identified. The major small RNA group identified was miRNAs.
Altogether, we
identified 298 miRNAs in bovine milk EXs. The effect of diets and milk processing
will be discussed.
16. Producing Hygienic Raw Milk: Standards, Testing, and Farmer Education
Joanne Whitehead, Independent Researcher, British Columbia, Canada
Joanne Whitehead1
, Nicholas Azzolina2
, Bryony Lake3
, Margaret Coleman4
1. Independent Researcher, British Columbia, Canada
2. Independent Consultant, Wisconsin, USA
3. Meta+ Research and Analysis, British Columbia, Canada
4. Coleman Scientific Consulting, Groton New York, USA
Evidence is accumulating from dairies around the world that counters the assumption
that raw milk is
inherently dangerous. Genomic research is providing insight into the importance of
milk�s diverse
microbiota, and large cohort studies are providing evidence for immunological
benefits of raw milk
consumption, especially during infancy. Can the risk of pathogens be reduced
sufficiently to take
advantage of the health benefits of raw milk?
Two recent independent analyses of the same dataset (US raw milk outbreaks reported
in 2005-2016)
reported similar findings, namely, that although legal access to raw milk in the US
has been increasing in
recent years, the rates of outbreaks, illnesses, and hospitalizations are not
increasing. Both analyses
accounted for legal status by state and population growth, as well as using raw
milk dairy licensing data
(where available) as a proxy for changes in consumption rates. One analysis
demonstrated the lack of
correlation between access to raw milk in the US and outbreak rates, and the other
demonstrated
through Poisson regression that the numbers of outbreaks, illnesses, and
hospitalizations within a state
57 | Page
Poster Presentation Abstracts
were not significantly related to the state legal status of raw milk. Is improved
farmer education
responsible for the increasing safety of raw milk?
Results from training provided to dairy farmers by the Raw Milk Institute (RAWMI)
suggest that
appropriate education contributes to safer milk. RAWMI trains farmers to develop
and maintain on-farm
hazard analysis and critical control points (HACCP)-based food safety systems, and
to verify controls
using microbiological testing and common standards. Data from Pennsylvania supports
this connection
between training and milk safety. RAWMI training provided at a workshop hosted by
Pennsylvania State
University�s College of Agricultural Sciences in 2014 likely contributed to the
subsequent decline in
numbers of raw milk outbreaks and illnesses in that state.
In a pilot study of herdshare milk in British Columbia, analysis of more than 160
milk samples produced
by farmers RAWMI-trained or in training documents non-detectable levels of
pathogens (E. coli STEC,
Campylobacter, Salmonella and Listeria) in all samples. Coliform counts from fully
trained farmers were
at or below 10 per millilitre in 96% of samples and never surpassed 40 cfu/ml,
while farmers in training
showed slightly higher frequency and maximum level of coliform contamination. In
all cases when more
than 10 coliforms per ml were detected, farmers were able to identify and eliminate
the source of
contamination.
Such food safety training programs incorporating microbiological testing and
management practices are
now accessible to farmers who wish to produce milk specifically for raw
consumption. Genomic research
is advancing on-farm food safety testing systems that can now provide farmers with
accurate and
inexpensive microbiology assay results within hours, providing data about potential
risks prior to sale.
Stringent hygienic standards developed specifically for producing raw milk for
human consumption,
specialized ongoing farmer training, and rapid on-farm testing, combine to reduce
raw milk risk
consistently and substantially. Further genomic characterization of microbes in
milk and broader
implementation of farmer education programs around the world should lead to even
further declines in
raw milk outbreaks and illnesses in the future.
17. Introducing �LIFE� - Pioneering Work in Human Milk Composition
Michael Affolter, Nestl� Research, Lausanne, Switzerland
Human milk constitutes the optimal nutrition for infants and is associated with
short- and long-term
health benefits. The WHO recommends exclusive breastfeeding during the first 6
month of life,
continued up to 24 month or beyond with adequate complementary feeding.
Understanding the composition of human milk and the factors affecting it is crucial
for the optimization
of maternal nutrition and health during pregnancy and lactation. As technological
advances take place,
scientists have gained more insights of the nutrients and bioactives in human milk.
LIFE (Lactation for Infant Feeding Expertise) is a dynamic research initiative
aiming i) to characterize
human milk composition along the lactation period and ii) to explore associations
with maternal
parameters as well as nutritional status, infant�s growth and other developmental
outcomes (Figure 1).
This program includes 16 clinical studies in 20 countries across the globe and uses
standardized
methods of milk sampling in different populations, including mothers of preterm and
small for
gestational age infants. Around 8000 human milk samples from more than 2000
participants at
different stages of lactation have been analyzed by state-of-the-art analytical
techniques. Twenty new
validated methods have been developed to analyze more than 200 nutrients and
bioactives in human
milk. Up to now, over 20 peer-reviewed scientific articles and reviews have been
published and more
are currently drafted.
58 | Page
Poster Presentation Abstracts
18. Effect of Milk Fat Globule Size on Growth and Bone Quality in Rats
Nurit Argov-Argaman, The Hebrew University of Jerusalem, Israel
Nurit Argov-Argaman1
, Hodaya Goldraich-Altman2 and Efrat Monsonego-Ornan2
1. Department of Animal Sciences, the Robert H Smith Faculty of Agriculture, Food
and Environmental Sciences. The
Hebrew University of Jerusalem, Israel.
2. Institute of Biochemistry and Nutrition, the Robert H Smith Faculty of
Agriculture, Food and Environmental
Sciences. The Hebrew University of Jerusalem, Israel.
Introduction: Milk fat is secreted in a unique structure termed milk fat globule
(MFG) which consists of
a triglyceride core covered with 3 layers of protein and polar lipid and termed MFG
membrane. The MFGs
are secreted in a wide range of sizes; from 200nm - 15�m. Size dependent
differences in MFG lipidome
and proteome were demonstrated in human, bovine and caprine milk and may determine
the bioactivity
of milk.
Recent studies showed that the MFG and its membrane exert positive health effects
for neonates and
adults possibly though protection against pathogenic bacteria and improved gut
microbiome.
The connection between the microbiome and bone growth is an emerging field, with
important
influence on both linear growth and bone mass. It has recently shown that the
microbiota of juvenile
mice sustains both weight gain and linear growth. Taken together, we hypothesized
that MFG and its
membrane may play a role in bone development, directly, or indirectly through the
gut microbiome. We
further hypothesize that MFG size may modulate the association between bone
development and
dietary MFG.
Methods: Three -weeks-old female Sprague-Dawley rats which were divided into 4
groups that were fed
on the experimental diets for 6 weeks; control group, receiving recommended diet
for rodents (AIN93), g high fat and low protein, vitamin and mineral diet group
(HFCN), HFCN drenched with large MFG
group (HFL, average diameter 3.5 �m, 0.275 gr/day) and HFCN diet drenched with
small MFG group (HFS,
average diameter 2.7 �m, 0.185 gr/day). Group food and water consumption were
monitored throughout
the experiment. Individual weight and length were monitored weekly. Plasma was
collected at the end
of the experiment as well as femurs from each rat, which were scanned by micro-
computed tomography
(mCT) for determination of geometric parameters.
Results: The control group consumed 125gr/day fat, while the high fat diet groups
(HFCN, HFL and HFS,
respectively) consumed 168, 200 and 205 gr/day of fat. While the control group�s
diet consisted of 51%
and 17% of polyunsaturated and saturated fatty acids respectively, the diet of the
high fat groups
consisted of 46% and 26% polyunsaturated and saturated fatty acids, respectively.
Feeding on high fat
diet reduced weight gain and length compared with control, whereas no differences
were found
between all 3 groups fed on high fat diets.
Food conversion efficiency was calculated for energy, protein and carbs. Improved
carb efficiency was
recorded when high fat diet was supplemented with MFG, and especially with small
MFG (HFS). All high
fat diets increased protein efficiency for growth, and the highest values were
found in the group
supplemented with small MFG, which was almost 2-fold higher compared with control.
These metabolic
differences were associated with differences in bone quality.
Discussion: This research suggests that MFG size has different effect on
development and growth in the
rapid growth phase in young rats. The different metabolic responses are manifested
by improve protein
efficiency for growth and femur bone characteristics in high fat and low protein
and micronutrients diet
supplemented with small MFG. the fact that no improvement in bone quality was
achieved when diet
was supplemented with large MFG, provide evidence for the size dependent metabolic
response for
MFG.
59 | Page
Poster Presentation Abstracts
19. Investigation of The Biological Effects of Milk Derived Extracellular Vesicles
on Cultured
Macrophages
Susanne Rossen Ascanius, Aarhus University, Aarhus, Denmark
Susanne Rossen Ascanius1
, Maria Stenum Hansen1
, Jan Trige Rasmussen1
1. Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark
Milk-derived extracellular vesicles (MEVs) may form a mother-to-infant
communication system
modulating the infants� immune system by its cargo, yet the mechanism behind is
unknown. This project
aims to reach a better understanding of how bovine MEVs affects the intestinal
immune system. The cell
line RAW 264.7 (mouse macrophages) was used as a model of the intestinal immune
system. Using
fluorescently labeled MEVs it was found that RAW 264.7 cells could take up MEVs.
The viability of the
cells was evaluated using an MTT cytotoxicity assay, showing no effects upon MEV-
addition, if the
incubation time was less than 12 h and the MEV concentration was below 100 �g
protein/mL. Lastly,
effects of MEV on cytokine secretion (IL-1�, IL-6, and IL-10) from LPS stimulated
RAW 264.7 cells were
measured using a sandwich ELISA. The first results indicated the secretion of IL-6
was downregulated
when RAW 264.7 cells incubated with MEVs for 4 h prior to a response provoked by
100 ng/ml LPS. Thus,
results agree with previous findings, but we seek to perform clarifying
experiments. The notion that
mother�s milk is able to modulate the immune response in infants by communication
through EVs is still
apt.
20. Allelic and Haplotypic Frequencies of Milk Protein Variants in French Bovine
Breeds
Didier Boichard, INRA-GABI, Jouy-en-Josas, France
Presented by Marie-Pierre Sanchez
M.-P. Sanchez1
, S. Fritz1,2, A. Delacroix-Buchet1
, D. Boichard1
1. GABI, INRA, AgroParisTech, Universit� Paris Saclay, F-78350 Jouy-en-Josas
2. Allice, F-75012 Paris, France
SNP chips make it possible to obtain a large number of genotypes at a reduced cost.
The chip used in the
Eurogenomics consortium includes most of the genetic variants responsible for the
major milk protein
variants. Used on a large scale for genomic selection purposes, it therefore
provides accurate results in
many breeds. In addition, these genotypes are imputed, i.e. missing markers are
predicted from family
and population genotypes for the same variants and for close markers. With these
very
reliable genotype predictions, it is possible to reconstruct information on milk
proteins for all animals
genotyped, even with older chips without the variants studied, and we can thus
estimate the trend of
allele frequencies over time. Finally, phases of alleles (haplotypes), i.e. their
distribution between
paternal and maternal chromosomes are determined during the imputation process. All
casein genes
are located in a small cluster of 250 kb and this haplotype information is
important because only some
combinations of alleles and thus a limited number of haplotypes do exist. This
poster presents the
frequencies of the main casein (CN) haplotypes and beta-lactoglobulin (�-LG)
alleles segregating in ten
French cattle breeds.
For �-LG, two variants (A and B) are frequent in all breeds. For caseins, only 11
haplotypes are present
(among 30 possible ones) in frequencies higher than 1% in at least one breed, and
each dairy breed
presents 4 to 7 haplotypes whereas beef breeds appear more diverse. The trends of
some allelic
frequencies over time will be presented on the poster.
60 | Page
Poster Presentation Abstracts
21. From�MIR: A Research Project to Predict and Study Milk Cheese-Making Properties
from MIR Spectra
in Montb�liarde Cows
Didier Boichard, INRA-GABI, Jouy-en-Josas, France
Presented by Marie-Pierre Sanchez
D. Boichard1
, V. Wolf2
, M.P. Sanchez1
, M. El Jabri3
, N. Gaudilli�re2
, E. Beuvier4
, S Fritz1,5, P. Trossat6
, S. Gavoye6
, Y.
Ga�z�re7
, E. Notz8
, M Brochard9
, P. Grosperrin2
, A. Delacroix-Buchet1
, C. Laithier3
1. GABI, INRA, AgroParisTech, Universit� Paris Saclay, Jouy-en-Josas, France
2. Conseil Elevage 25-90, Roulans, France
3. Institut de l�Elevage, Paris, France
4. URTAL, INRA, Poligny, France
5. Allice, Paris, France
6. ACTALIA, Poligny, France
7. EnilBio, Poligny, France
8. Centre Technique des Fromages Comtois, Poligny, France
9. Umotest,Ceyz�riat, France
From�MIR is a research project conducted in Montb�liarde cows to predict milk
cheese-making
properties (CMP) from mid-infrared (MIR) spectra in order to control them through
herd management
and breeding. The Montb�liarde breed represents about 20% of French dairy cows and
in its region of
origin (Franche Comt�, Eastern France), it is mainly bred for high quality cheese
production. From�MIR,
carried out between 2015 and 2018 and funded by Casdar, Cniel and Urfac, was
managed by CEL25-90
milk recording organization and has involved many partners (Umotest, CEL70, Eva
Jura, Actilait, EnilBio,
Idele, Allice and INRA). To establish the MIR prediction equations, 420 milk
samples were collected under
a wide range of conditions (in herds throughout the region, in summer and winter,
in different lactation
stages, in evening and morning milkings, from animals with various lactoprotein
genotypes, etc.). For
each sample, 24 reference measurements of coagulation, cheese yield and
acidification were performed
for either soft cheese or pressed cooked cheese. MIR spectra were recorded and then
standardized
using Grelet�s method. For prediction equation development, the Partial Least
Squares method with
variable selection proved to be more accurate than the Bayesian methods tested
(Bayes A, B and C). Of
the 24 CMP, nine were predicted with medium to excellent accuracy (R2 from 0.59 to
0.92). These 9
traits related to yield (3), coagulation (5) and acidification (1) were then
predicted on a large scale for
about six million spectra collected from more than 400,000 cows in the Franche
Comt� region between
2011 and 2017. Based on these data, the main environmental factors were identified
(in particular herds,
parity, lactation stage and feeding level) and a large panel of genetic analyses
was conducted. The nine
CMP traits presented medium to high heritabilities at the test-day level (from 0.37
to 0.48) and their
genetic determinism was the same across lactations and all along the lactation
(except in the first month
of lactation, which is significantly different). Moreover, CMP traits were
favorably correlated between
them, positively correlated with fat and protein contents, negatively correlated
with milk yield, and
independent from other traits included in the breeding objective. The 50k
genotypes, available for about
20,000 cows, were imputed at the whole genome sequence level and we identified 59
genomic regions
(QTLs) associated with CMP and a network of 736 genes functionally related to CMP
(Sanchez et al, this
seminar). Finally, good accuracies of genomic predictions, estimated including QTL
and 50k marker
effects, were found (R2=0.65 on average for all CMP traits). Genetic trends,
estimated over the last 15
years, seemed favorable, probably due to the high weight given to protein and fat
content in the
breeding goal of the Montb�liarde breed. However, we have also shown that directly
including CMP in
the breeding goal could improve milk cheese-making traits more efficiently. All
these results led to the
implementation of a genomic evaluation of CMP predicted from MIR spectra in
Montb�liarde cows and
are already of interest for other French bovine dairy breeds or ruminant species.
61 | Page
Poster Presentation Abstracts
22. Metabolic analysis of the gut microbiota using an in vitro platform, and its
future application for
studies on cow�s milk
Jenni Firrman, Eastern Regional Research Center, ARS, USDA, Wyndmoor, Pennsylvania,
USA
Jenni Firrman1
, Peggy Tomasula1
, LinShu Liu1
, Ceylan Tanes2
, Elliot Friedman3
, Kyle Bittinger2
, Scott Daniel2
, Pieter Van
den Abbeele4
, Bradley Evans5
1. Dairy and Functional Foods Research Unit, Eastern Regional Research Center, ARS,
USDA, Wyndmoor, PA, USA
2. Division of Gastroenterology, Hepatology, and Nutrition, Children's Hospital of
Philadelphia, Philadelphia, PA, USA
3. Division of Gastroenterology, Perelman School of Medicine, University of
Pennsylvania, Philadelphia, PA, USA
4. ProDigest, Ghent, Belgium
5. Donald Danforth Plant Science Center, Proteomics & Mass Spectrometry Facility,
St. Louis, MO, USA
Cow�s milk is a complex food and a source of high-quality proteins, fat,
carbohydrates, calcium and other
minerals, and vitamins, that are associated with a number of positive health
outcomes in children and
adults. Recently, research on the digestion of HTST pasteurized and homogenized
whole milk was
performed utilizing a simulated gastrointestinal model. Although most of the milk
proteins were
completely digested by the end of the process, peptides of about 5 to 10 kDa in
size remained. A partial
analysis of the sample for casein phosphopeptides (CPP), a known milk bioactive,
found that 31 CPP were
released during intestinal digestion. Fat globules were present throughout the
digestion process and
coalesced into larger globules that were stabilized by proteins. Based on these
results it is expected that
after milk consumption, multiple digested components will pass through the small
intestine and enter
the colon. Here they will interact with the gut microbiota, a large population of
bacteria composed of
500-1000 species, intimately associated with human health and disease and altered
by dietary changes.
However, there is limited knowledge regarding the effect of bovine milk on the gut
microbiota in terms
of changes to the community structure and metabolic functions. In future studies,
the effects of nonfat
milk and whole milk, and other milk components on the gut microbiota will be
elucidate through
application of the Twin Simulator of the Human Intestinal Microbial Ecology
(TWINSHIME).
The TWINSHIME is an in vitro system composed of individual bioreactors designed to
reproduce the
conditions of the entire gastrointestinal tract from the stomach to waste. As a
TWINSHIME�, there are
two complete systems set up to run in parallel. Each system has individual
bioreactors to represent the
stomach and small intestine, and divides the colon into the ascending, transverse,
and descending
regions. Previously, the system was evaluated for its ability to produce a stable
bacterial community.
Here, this work was expanded to analyze the metabolic profiles of the colon regions
using targeted and
untargeted metabolomics based on gas and liquid chromatography and mass
spectrometry.
Comparative analysis using Pairwise Pearson�s Correlation revealed a large
divergence in the metabolic
profiles of the AC to the TC and DC regions. This was confirmed using Principal
Components Analysis,
which demonstrated that the maximal distance between the AC and TC/DC samples was
about 24%. Total
short-chain fatty acid production increased from the AC to the TC to the DC
regions, 84.17, 89.57, 113.68
mMol/L, respectively. Bile acid analysis found that primary bile acids were at
higher concentrations in
the AC region and converted to secondary species in the TC and DC colon regions.
This data was then
coupled with metagenomic sequencing, which revealed that the regionally derived
metabolic profiles
had strong correlation to community composition and genetic potential based on
Procrustes analysis.
This was confirmed with a Mantel test that found r=0.8231 and p= .[Link] results
of this study provided
key insights regarding the metabolic divergence of the regional communities that
are integral to
understanding the structure-function relationship of the gut microbiota. This
knowledge is paramount
to the application of this system because it provides critical information on the
interactions between the
regional gut microbial communities and nutrients. This information will be applied
to studies using the
TWINSHIME to analyze the intereffects of cow�s milk, and cow�s milk components with
the gut
microbiota.
62 | Page
Poster Presentation Abstracts
23. The Whereabouts and Bioavailability of Vitamin B12 in Milk
Christian W. Heegaard, Aarhus University, Aarhus, Denmark
Presented by Sara Fris Rasmussen
Sergey N. Fedosov1
, Eva Greibe2
, Namita Mahalle3
, Vijayshri Bhide3
, Marie S. Ostenfeld4
, Lars Holm5
, Sadanand Naik3
,
Ebba Nexo2
, Christian W. Heegaard1
1. Department of Molecular Biology and Genetics, Aarhus University, Denmark
2. Department of Clinical Biochemistry, Aarhus University Hospital, Denmark
3. Department of Pathology, Deenanath Mangeshkar Hospital and Research Center,
Pune, India
4. Arla Foods Ingredients Group P/S, Denmark
5. School of Sport, Exercise and Rehabilitation Sciences, University of Birmingham,
United Kingdom
Vitamin B12 (B12) deficiency is of concern amongst vegetarians, infants, pregnant
women, and the
elderly. Animal products and byproducts are the only natural source of B12 in human
diets. Among those,
epidemiological studies and intervention trials suggest that milk and milk products
are attractive sources
of B12.
Here we summarize our results on B12 forms and distribution in milk as well as in
vitro, animal and human
studies comparing the effect of milk/milk fractions and the form of B12 present in
milk with B12 form
present in vitamin pills. We conclude that milk and milk products are valuable
sources of B12.
B12 in cow�s milk consist mainly of hydroxo-B12 (HO-B12) (4 �g/L � twice the
recommended dietary
allowance (RDA)). In contrast, the artificial cyano-B12 (CN-B12) is predominantly
employed in vitamin
supplements and food fortification. Half of B12 present in milk is in the whey
fraction bound to
transcobalamin (TC) and half is bound to the casein micelles.
TC promotes the transport of B12 through the intestinal cell line CaCo2. However,
free B12 is absorbed
as efficiently as is TC-B12, when examined after oral feeding to rats.
Interestingly we observe major
differences between the behavior of HO-B12 (present in milk) and CN-B12 (present in
vitamin pills). The
two forms are absorbed alike, however HO-B12 is transported more efficiently to the
liver than is CN-B12,
reflected also by a higher accumulation of CN-B12 in the circulation of the
experimental animal. In
addition, a large part of CN-B12 remains unchanged, while HO-B12 is converted to
the active coenzyme
forms of the vitamin.
In B12 deficient humans, equivalent amounts of B12 administrated as two servings of
CN-B12 capsules,
milk, or whey powder isolate (WPI) for up to eight weeks are equally efficient in
improving metabolic
biomarkers of B12 deficiency. As for the animal studies supplementation, CN-B12
supplementation
results in a higher increase in plasma B12 than does supplementation with HO-B12.
The finding
underscores that plasma B12 is a poor marker for judging the effect of
supplementation with B12.
We observed a beneficial effect on B12 biomarkers in healthy elderly receiving a
daily serving of whey
powder isolate (WPI) for one year. In contrast, the control group that received an
equicaloric
carbohydrate supplement showed a decline in the B12-biomarkers. Our observations
suggest a need for
B12-supplementation, even among elderly well ahead of the RDA for B12.
In conclusion, our findings underscore the benefit of improving B12 status by
administration of dairy that
supplies HO-B12. B12 in milk and WPI is absorbed as efficiently as is CN-B12 but is
superior as judged from
tissue distribution and conversion to the coenzyme forms of B12.
63 | Page
Poster Presentation Abstracts
24. Determination of Plasma Phospholipids Following Intake of Different Dairy
Matrices: A Randomized
Crossover Intervention Study
Emilie Ingvordsen Lindahl, Aarhus University, Aarhus, Denmark
Emilie Ingvordsen Lindahl1
, Louise Kj�lb�k2
, Jesper Malling Schmidt1
, Emilien Rouy3
, Klaus Juhl Jensen3
, Arne Astrup2
,
Mette Bakman3
, Kristian Wejse Sanggaard4
, Marianne Hammersh�j1
, Anne Raben2
, Hanne Christine Bertram1
1. Department of Food Science, Aarhus University, Aarhus, Denmark
2. Department of Nutrition, Exercise and Sports, University of Copenhagen,
Copenhagen, Denmark
3. Arla Foods Amba, Aarhus, Denmark
4. Arla Foods Ingredients, Aarhus, Denmark
Dyslipidemia in the fed state is a known contributor of cardiovascular disease
(CVD), and metabolic risk
factors such as obesity, insulin resistance and diabetes mellitus type 2 are all
associated with aberrant
circulating lipid profiles[1].
After intake of a meal, dietary phospholipids are absorbed and incorporated into
lipoproteins, including
chylomicrons and high density lipoprotein (HDL)[2,3]. The majority of dietary
lipids are hydrophobic and
thus transported in the circulation by lipoproteins. Levels of low density
lipoprotein (LDL) and
lipoprotein remnants [1,4] are thought to be directly associated with CVD risk,
however, the link between
postprandial phospholipids, their absorption kinetics and CVD risk is poorly
understood. Nonetheless,
disturbances in circulating lysophosphatidylcholine levels [5] as well as the ratio
of
phosphatidylcholine:phosphatidylethanolamine [6] and decreased ether-phospholipids
[7] are seen in
obesity and/or insulin resistance.
Evidence is emerging that the food matrix contributes considerably to the
absorption kinetics of food
components and thereby their nutritional properties [8]. Dairy is a heterogeneous
food group consisting
of products with various structural and textural configurations, and accumulating
findings indicate that
the dairy matrix can modulate postprandial lipidemia [9,10]. Therefore, the present
study investigates the
impact of the dairy matrix of four nutrient-matched dairy products with different
textural and structural
properties on postprandial phospholipid absorption kinetics.
The study was conducted as a randomized crossover design, consisting of four
different interventions;
cheddar cheese (i.e. intact protein network and milk fat globules); homogenized
cheddar cheese (i.e.
loss of protein network and potential alterations to the milk fat globules); MCI
drink (micellar casein
isolate (MCI) with added cream); MCI gel (gel product made from the MCI drink).
Each test day was
separated by a wash out period of minimum 2 weeks in-between. Twenty-five healthy,
normal weight
male subjects (aged 19-40 years) were recruited. The subjects received four
isocaloric, macronutrientmatched test meals, consisting of one of the dairy
products, bread and water. Blood samples were
drawn at time 0 min (fasting), as well as at 30, 60, 90, 120, 180, 240, 300, 360,
420 and 480 min after
ingestion of the test meals.
Using a targeted liquid chromatography tandem mass spectrometry method,
quantification of
approximately 50 glycerophospholipid and sphingolipid species will be performed,
potentially providing
new knowledge about the effects of the dairy matrix on postprandial phospholipid
status.
The present study is part of the interdisciplinary DAIRYMAT project, investigating
the structural and
textural properties of the dairy matrix on postprandial lipidemia.
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Poster Presentation Abstracts
25. Interrelations Between Chemical Changes in Lactose-Free UHT Milk
Lotte Juul Knudsen, Aarhus University, Tjele, Denmark
L. J. Knudsen1
, S. D. Nielsen1
, V. Rauh2
, D. Otzen3
, P.J.T. Dekker4
, L. B. Larsen1
1. Aarhus University, Department of Food Science, Tjele, Denmark
2. Arla Foods Innovation Centre, Agro Food Park 19, Aarhus, Denmark
3. Aarhus University, Interdisciplinary Nanoscience Center, Aarhus C, Denmark
4. DSM Biotechnology Centre, Delft, the Netherlands
Lactose free milk is an important dairy alternative for lactose intolerants.
However, lactose-free ultrahigh temperature (UHT) milk has a shorter shelf life (6
months) than conventional UHT milk (9 months),
which challenges export and product distribution for the food industry, as well as
sensory qualities in
relation to consumers. The shorter shelf-life is linked to a higher reactivity of
the lactase generated
galactose and glucose as substrate for the Maillard reaction compared to lactose.
Moreover, lactase
preparations with different purities may differently affect the Maillard reaction.
Furthermore,
proteolytic activities in lactose-free milk can lead to bitter peptide formation
and, in pathways either
involving Maillard reactions or not, may also lead to protein cross-links and
aggregation. These processlinked changes can contribute to accelerated product
deterioration in lactose-free UHT milk. Therefore,
the overall goal of this project is to improve shelf-life and quality of lactose-
free UHT milk. In the study,
three new lactase enzymes, which differ in purities and activities, will be
characterized. Different lactase
enzymes will be added to UHT-treated whole milk, either pre- or post-hydrolysis.
The milk samples will
be stored for up to one year at either 25 or 35 �C. Mass spectrometry and different
protein analytical
methods will be used to investigate proteolytic activities, Maillard reaction,
cross-links, aggregate
formation and inter-relations between these chemical changes. To study the Maillard
reaction and
process induced cross-links in relation to aggregation, a new mass spectrometry
(MS) single step
method in MRM mode using Triple Q MS has been developed to quantify markers of
these reactions
(furosine, N-carboxymethyllysine (CML), N-carboxyethyllysine (CEL), lysinoalanine
(LAL) and lanthionine
(LAN)).
26. DGAT1 K232A Polymorphism and Feeding Modify Milk Fat Triacylglycerol
Composition
Sara Pacheco, Wageningen University, Wageningen, The Netherlands
Sara Pacheco Pappenheim1
, Sine Yener1
, Hein J.F. van Valenberg1
, Daylan A. Tzompa-Sosa 1,3, Henk Bovenhuis2
1. Dairy Science and Technology Group of FQD, Wageningen University, Wageningen,
The Netherlands
2. Animal Breeding and Genomics, Wageningen University, Wageningen, The Netherlands
3. Laboratory of Food Technology and Engineering, Department of Food technology,
Safety and Health, Faculty of
Bioscience Engineering, Ghent University, Ghent, Belgium
The present study aimed to investigate the changes in TAG composition as affected
by alterations in
cow�s diet due to the seasonal variations and genetic factors. For this study 50
milk fat samples in winter
and 50 in summer were used from 25 cows with the DGAT1 KK genotype and 25 cows with
the DGAT1 AA
genotype. The samples were analyzed for milk fat content (%), fat composition and
TAG composition.
We found that, compared to winter, in summer the TAG species CN54 were higher and
CN34 and CN36
were lower. This seasonal variation in TAG profile was related to the seasonal
changes in the FAs C14:0,
C16:0, C18:0, C18:1cis9, total UFA and total LCFA most likely resulting from
dietary differences between
seasons. Furthermore, the effect of DGAT1 K232A polymorphism on the TAG profile was
quantified and
there was a significant effect on the TAG species CN36 with higher values for the
DGAT1 KK genotype.
When adjusting for differences in fat content there were no significant effects of
the DGAT1 K232A
polymorphism on the TAG profile. A significant interaction between the DGAT1 K232A
polymorphism and
season was detected for TAG species CN42 and CN52; in summer the DGAT1 KK genotype
was associated
with higher levels for TAG species CN42 than the DGAT1 AA genotype whereas in
winter the difference
between the genotypes was small. For CN52 the DGAT1 AA genotype was associated with
higher levels
in summer than the DGAT1 KK genotype. In winter the difference between the
genotypes was also small.
We show that regardless of the preference of the DGAT1 genotype (AA/KK) and
depending on the
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Poster Presentation Abstracts
availability of FA according to season, UFA (C18:1cis9),SCFA (C6:0 and C10:0) and
MCFA might be
esterified on the glycerol backbone of the TAG, keeping the structure
characteristics of each TAG
species. To our knowledge, this is the first report on the interaction effect of
DGAT1 K232A polymorphism
and season on the TAG composition in MF.
27. Natural Variations in Danish Dairy Milk Components - Technological and Health
Related Aspects
Natacha Roed R�in, Aarhus University, Tjele, Denmark
N. R. R�in1
, J. W�rts1
, N. A. Poulsen1
, L. B. Larsen1
1. Aarhus University, Department of Food Science, Tjele, Denmark
The inherent variation of milk represents both opportunities and challenges for the
dairy industry. The
development of specialized products for different markets with different needs is
highly influenced by
the quality of milk as a raw material. The inherent variation of milk facilitates
the opportunity for a better
differentiation of dairy ingredients and hence the development of new products.
However, it also
represents challenges regarding processing variation. An increasing amount of the
milk produced is used
for ingredients involving processes like up-concentrations, isolations and
filtration steps, with
subsequent enlargement of the variance experienced.
The DanMilk project has the overall aim to map Danish dairy milk and to clarify the
significance of
different variation factors (season, geography, breed and management system) in
relation to a large
range of milk components [1][2]. Raw silo milk is collected from 15 dairies
representing 33 different raw
milk streams of silo milk every month over a two-year period. The milk samples are
subjected to
profound studies of the milk composition (fat, protein, vitamins and elements,
metabolites, somatic cell
count and germ number). As a part of DanMilk, a PhD project will focus on the
implementation and use
of analytical techniques for the profiling and quantification of specific
compounds, including the detailed
milk protein composition and its natural post translational modifications (PTMs)
[3], as well as
indigenous enzymes in relation to differential cell count[4]. The indigenous
enzymes cover the plasmin
system and cathepsins, which will be determined by combinations of enzymatic assays
as well as by
immunological methods in combination with mass spectrometry. The proteolytic
enzymes can play roles
in both technological properties of the milk, as well as in e.g. the formation of
bioactive peptides, and in
physiological processes in the udder. An example of a metabolite of interest is
phytoestrogens, which is
known to vary in milk with e.g. grass feeding, and these will be determined by
LC/ESI-MS.
Overall, the project aims to provide a solid understanding and knowledge of the
level of natural variances
in specific milk components and their implications for upstream dairy processes and
related health and
nutritional aspects.
28. Mechanistic Background for the Beneficial Effects of Osteopontin and
Caseinoglycomacropeptide
Esben Skipper S�rensen, Aarhus University, Aarhus, Denmark
Brian Christensen1
, Karoline Rusbjerg S�rensen1
, Marie Rytter S�rensen1
, Anette M�llertz2
, Scheyla Daniela J�rgensen2
,
Lotte Neergaard Jacobsen3
, Marie Stampe Ostenfeld3
, Esben Skipper S�rensen1
1. Department of Molecular Biology and Genetics, Aarhus University, Denmark
2. Department of Pharmacy, Copenhagen University, Denmark
3. Arla Foods Ingredients, Viby, Denmark
Intervention studies have indicated beneficial effects of the milk components
osteopontin (OPN) and
caseinoglycomacropeptide (CGMP) on intestinal health and development of the immune
system.
However, the underlying mechanism for these effects remains unknown. To induce
physiological
effects, the components must resist gastrointestinal transit in a bioactive form.
Therefore, we have
subjected OPN and CGMP to in vitro simulated gastrointestinal transit and analyzed
the effect of the
generated peptides in model systems. For OPN, we found that the fragment Trp27-
Phe151 containing the
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Poster Presentation Abstracts
integrin binding RGD-sequence resisted in vitro gastric digestion. This resistance
was dependent on
glycosylation of threonine residues
near the integrin binding sequence in both human and bovine milk OPN. The fragment
was capable of
crossing an in vitro model of the intestinal barrier and it retained ability to
interact with integrins in an
RGD-dependent process. These results suggest a mechanism for how ingested milk OPN
can induce
physiological effects via integrin signaling in the intestine or even systemically
by uptake over the gutblood barrier.
For CGMP, we have shown that digested CGMP increases the integrity and tightness of
a Caco-2 cell
model of the intestinal barrier. Moreover, we have modelled intestinal inflammation
by addition of the
inflammatory cytokines IL-1�, TNF-a, interferon-? and LPS to the Caco-2 cell model.
In this model, CGMP
was found to mitigate the induced inflammation. In addition, digested CGMP reduced
LPS and TNF-a
induced secretion of the inflammatory marker IL-8 in intestinal epithelial cells in
vitro. Collectively, these
data suggest molecular mechanisms for the beneficial effects of CGMP reported in
clinical studies of e.g.
inflammatory bowel diseases.
In conclusion, our results suggest molecular mechanisms by which the bioactive milk
proteins OPN and
CGMP can exert the beneficial functions shown in clinical studies.
29. Effect of Genetic Variants if ?-Casein on the Digestibility of Milk Protein
Bulei Sheng, Aarhus University, Tjele, Denmark
Bulei Sheng1
, S�ren Drud-Heydary Nielsen1
, Nina Aagaard Poulsen1
, Lotte Bach Larsen1
1. Aarhus University, Department of Food Science, Tjele, Denmark
Ingestion of dairy products has been associated with health-related effects coming
from genetic
polymorphisms of milk proteins. ?-casein has different genetic variants resulting
in amino acid
substitution and displays varying degree of glycosylation, like e.g. ?-casein
genetic variant B is more
glycosylated compared to variant A. Both genetic variants and glycosylation degree
may affect
macromolecular structures in the milk, like the organization of the casein
micelles, which may influence
the rheology behavior and hydrolysing rate of casein during gastric digestion. When
ingested, dairy
proteins are exposed to a complex array of digestive proteases, and the proteins
will release a source of
peptides, which may exert different biological functions. Yet, little work has
explored how natural
variation, including glycoisoforms and genetic variants, will influence the
digestibility of milk protein and
affect the release of glycopeptides.
Recently, 11 individual milk samples were obtained from Holstein cows to prepare
pool milk with same
genetic variants for ?-casein, AA (n = 3), BB (n = 4) and AB (n = 4). Afterwards,
the casein micelle size was
measured by dynamic light scattering (Nano-flex). Furthermore, digestion of the
pooled skim milk
samples was performed by the modified in vitro static digestion model based on
INFOGEST 2.0, followed
by fluorescamine assay and SDS-PAGE to characterize the digestibility of major milk
proteins. A tendency
towards a lower digestion rate was observed for milk originating from ?-casein BB
and AB cows during
the gastric phase, and a decrease in their hydrolysis degree after intestinal
digestion compared to AA.
This may be related to a higher degree of glycosylation of ?-casein for BB and AB
milk samples compared
to AA. For next step, Tris-Tricine gel and LC-MS single Q will be applied for
peptidomics study of the
digests.
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Poster Presentation Abstracts
30. Plasmin-induced Hydrolysis on as1-caseins with Eight and Nine Phosphate Groups
(as1-CN-8P/9P) and
�-casein with Two Genetic Variants (�-CN-A1/A2) in Model Cheeses During Ripening
Huifang Cai, Wageningen University, Wageningen, The Netherlands
Huifang Cai 1,2, Etske Bijl2
, Elke Scholten1
, Guido Sala1
1. Physics and Physical Chemistry of Foods, Wageningen University, the Netherlands
2. Dairy Science and Technology Group, Food Quality and Design, Wageningen
University, the Netherlands
Natural variations of milk components, especially proteins and native enzymes, are
known to have a
marked influence on proteolysis and texture development during cheese ripening.
However, the
knowledge of these variations in milk composition and of their influence on cheese
texture is still
incomplete. More studies in this area can provide tools to better control cheese
texture, allowing the
optimization of existing products and the creation of new ones.
Plasmin, a primary indigenous enzyme, plays a crucial role in casein degradation
and texture
development in many cheeses with high scalding temperature, such as Parmesan.
Recently, it has been
reported that as1-casein variants with different phosphate groups (as1-CN-8P/9P)
show different
efficiency in degradation by chymosin in model milk gels during ripening, as well
as �-casein with two
genetic variants (�-CN-A1/A2). However, plasmin-induced hydrolysis of specific
variants of casein and its
effects on cheese texture changes during ripening are less studied.
The aim of this study was to investigate plasmin-induced hydrolysis of as1-CN-8P/9P
and �-CN-A1/A2 and
to relate the observed proteolytic patterns to texture changes during ripening in
model cheese. Four
batches of cheese with different plasmin addition (0-1.0 �l plasmin/ L milk) were
made and stored for 12
weeks at 16�C. Our results suggest that plasmin-induced hydrolysis of as1-CN might
be affected by
variations in phosphorylation. About 31% more as1-CN-9P than as1-CN-8P was
hydrolyzed in cheese with
additional plasmin (1.0 �l/ L milk). However, differences in degradation of as1-CN-
8P/9P were less
significant in model cheese with lower plasmin activity. Probably, small changes in
phosphorylation
influence the accessibility of these proteins by plasmin and consequently cause
differences in casein
degradation efficiency. In addition, no significant differences were found for
plasmin-induced hydrolysis
of different genetic variants of �-CN. For cheese texture properties, intact as1-
CN-9P fraction showed
higher correlations with resilience and cohesion when compared to as1-CN-8P. These
findings represent
a starting point for further studies about the impact of natural variations in milk
composition on
proteolysis and cheese texture development during ripening.
31. Comparative Peptidomic Analysis of Human Milk Reveals Specific Enzyme-Substrate
Interactions
Junai Gan, University of California-Davis, CA, USA
Junai Gan1
, Jingyuan Zheng2
, Nithya Krishnakumar1
, Elisha Goonatilleke3
, Carlito B. Lebrilla3,4, Daniela Barile1,4, J. Bruce
German1,4
1. Department of Food Science & Technology, University of California, Davis, CA,
USA
2. Department of Nutrition, University of California, Davis, CA, USA
3. Department of Chemistry, University of California, Davis, CA, USA
4. Foods for Health Institute, University of California, Davis, CA, USA
Human milk contains endogenous enzymes that together with milk proteins comprise a
dynamic
proteolytic system; yet, biological properties of these enzymes, their targets, and
their activities in
response to variations within infants are unclear. The pH variations in the infant
digestive system can
lead to distinct molecular changes of human milk, which subsequently can stimulate
different
physiological responses. Deciphering the health impacts of such complex
physiological processes
requires a mechanistic understanding of how infant conditions affect these
molecular changes within
milk. This study was designed to determine the specificity of enzyme-substrate
interactions in human
milk as a function of pH. Freshly expressed human milk was incubated at
physiologically relevant pH�s
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Poster Presentation Abstracts
ranging from 2 to 7, and peptides were extracted and analyzed by liquid
chromatography coupled with
tandem mass spectrometry (LC-MS/MS). To characterize the pattern of milk
proteolysis, a heatmap was
generated with z-scores of sequence occurrences centered and standardized by each
protein, and the
clustering of proteins was based on Pearson correlation. The index of dispersion or
variance-to-mean
ratio of sequence occurrences in different groups was calculated for each protein.
Comparative analysis
revealed that effects of pH on proteolysis are distinct among human milk proteins.
Ranked by the index
of dispersion (or variance-to-mean ratio) of sequence occurrences across groups, a-
lactalbumin was
most sensitive to the changes of pH. Further analysis and experiment revealed that
the endogenous milk
enzyme cathepsin D cleaves a-lactalbumin to release specific peptides as the milk
pH shifts from 7 to 3.
Integrating clinical, analytical, biochemical, bioinformatical and statistical
resources, this approach
enabled us to gain insights into the endogenous proteolysis of human milk from the
omics data sets.
Controlled proteolysis of milk can guide innovations towards precision design of
infant feeding and
change the value proposition of milk for targeted health management.
32. Human Breast Milk Metabolomes Over the Course of 6 Months Lactation
Katrine Overgaard Poulsen, Aarhus University, Aarslev, Denmark
Katrine Overgaard Poulsen1,2, Fanyu Meng3
, Elisa Lanfranchi3
, Alan L. Kelly3
, and Ulrik K. Sundekilde1
1. Department of Food Science, Aarhus University, Aarslev, Denmark
2. Sino-Danish Center for education and research (SDC)
3. School of Food and Nutritional Sciences, University College Cork, Cork, Ireland
Background: The complexity and dynamic nature of human breast milk (HBM) have long
been
acknowledged and deemed essential for infants� health and growth during the first
period of life.
Improvements in analytical techniques in recent years have allowed for a more
comprehensive
understanding of the composition of HBM in terms of minor constituents as
metabolites, as well as
microorganisms. Interconnections between the human breast milk metabolome (complete
set of
metabolites found in the breast milk at a given time point), oligosaccharides and
microorganisms, are
not well studied, and the impact of associations on the health and maturation of
the respective infant
less understood. The Infamilk study explores this interface via milkomics,
involving conducting
proteomics, metabolomics and microbiome sequencing on HBM of mothers delivering at
term and preterm.
Methods: The present sub-study covers metabolomics analysis of HBM and describes
variation of HBM
metabolites by nuclear magnetic resonance (NMR) based metabolomics from mothers
delivering at
term. Human breast milk samples from mothers at four different time-points
following birth derived
from Infamilk are analyzed. In total, 100 samples from the time-points week 1 (W1),
week 4 (W4), week
8 (W8) and week 24 (W24) are included, and the respective metabolome identified and
quantified. 1H
NMR spectra acquisition was conducted by the use of a Bruker Avance III 600 MHz
spectrometer with a
frequency of 600.13 MHz containing a 5 mm 1H TXI probe (Bruker BioSpin,
Rheinstetten, Germany).
Expected results: Representative metabolic profiles of HBM over the course of 6
months lactation are
presented, which ultimately will be compared to the breast milk microbiota of the
mothers (separate
study). Thus, by integrating the acquired knowledge of milkomics analyses,
complementary
compositional and functional analyses allows not only for expanding the scientific
area of HBM, but also
for potentially optimizing infant formulae.
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