Protecting flaxseed from ruminal biohydrogenation increases the omega3

Protecting flaxseed from ruminal biohydrogenation increases the omega-3 profile in cow’s milk Randi Wilson 1, Sarah Akers 2, Katherine Swanson 2, Gita Cherian, Ph. D. 2, & Gerd Bobe Ph. D. 2 Bio. Resource Research Interdisciplinary Program 1 Department of Animal & Rangeland Sciences 2 Oregon State University Corvallis, OR

Milk is the most affordable, nutrient dense food available to humans • Top dietary source of (Drewnowski, 2011): • Retinol/Vitamin A • Vitamin D • Calcium • Potassium • Second most important dietary source of (Drewnowski, 2011): • Riboflavin/Vitamin B 2 • Phosphorus • Third most important dietary source of (Drewnowski, 2011): • Zinc • Colbalamin/Vitamin B 12 • Selenium

(Drewnowski, 2011)

Milk Consumption has been on the decline for the past decade Annual Milk Consumption in the United States from 2005 - 2015 190 Milk Consumption (lbs person) 185 180 175 170 165 160 155 150 2005 2006 2007 2008 2009 2010 2011 Time (years) 2012 2013 2014 2015 (USDA ERS - Dairy Data)

Milk consumption is declining due to certain health implications • Milk fat is comprised of (Chen et al. , 2004): • 66% Saturated Fatty Acids (SFAs) • 30% monounsaturated fatty acids (MUFAs) • Only 4% polyunsaturated fatty acids (PUFAs) Low Density Lipoproteins (LDL) Cholesterol Chance of Plaque Formation Risk of Cardiovascular Disease, obesity and diabetes (Moallem et al. , 2012)

Increasing PUFAs can improve health qualities of milk • Milk fat is comprised of (Chen et al. , 2004): • 66% Saturated Fatty Acids (SFAs) • 30% monounsaturated fatty acids (MUFAs) • Only 4% polyunsaturated fatty acids (PUFAs) α – linolenic acid Risk of CVD, Hypertension, & Diabetes (Alvarado-Gilis et al. , 2015) Promotes normal growth, brain development, vision, and immunity (Resende et al. , 2015)

Feeding flaxseeds to cows can improve the fatty acid profile of the milk • Naturally high in: A previous study did not see a significant change, but they did • Essential omega-3 fatty acids notice a trend. (Oeffner et al. , 2013) • PUFAs • Specifically α – linolenic acid (ALA) aka an omega-3 fatty acid • SFAs overall ©Sean Dreilinger (Oeffner et al. , 2013) (Resende et al. , 2015)

Cows are ruminant animals 4 – Chambered “Stomach” 1. Reticulum 2. RUMEN 3. Omasum 4. Abomasum ©USDA

CH 4 H 2 CO 2 To Mouth Gas layer Forage Layer Fluid and finer particles which will move on to the omasum followed by the abomasum To Omasum/Abomasum

Ground Flaxseed or Extruded Flaxseed Mammary PUFAs SFAs Glands (Polan et al. , 1964)

α-linolenic acid C 18: 3 cis 9, 12, 15 Isomerase C 18: 3 cis-9, -15, Reductase trans-11 C 18: 2 cis-15, trans-11 Reductase Ruminal Biohydrogenation Stearic Acid C 18: 0

A new method of processing flaxseed called rumen-protection is proposed to prevent ruminal biohydrogenation of omega-3 FAs Hypothesis: This rumen-protected flaxseed supplement will successfully help omega-3 FAs bypass the rumen, allowing it to be incorporated into milk in the mammary gland. This will increase omega 3 FAs and decrease overall SFAs in the milk

2 Weeks Rumen-bypass Flaxseed 2 Weeks Bl Bl oo oo d d TMR Only Ground Flaxseed Blood Rumen-bypass 2 Weeks Flaxseed TMR Only 2 Weeks Bl oo oo Bl End of Study d d 2 Weeks d Ground Flaxseed 2 Weeks 2 Blood Weeks d R only TM 2 Weeks Bl oo d Rumen-bypass 2 Weeks Flaxseed Ground Flaxseed

University of Tennessee: Institute of Agriculture

Crude Blood/Milk Sample Extraction of Lipids ©ACOS Lipid Library © User: Offnfopt / Wikimedia Commons / CC-BY-SA-3. 0

Rumen-protected flaxseed does not impede significantly on productions and health parameters • Milk Yield • Dry Matter Intake • Milk components not altered with the exception of lactose • No significant changes in serum metabolites with the exception of lower beta-hydroxybutyrate concentrations

Rumen-protected flaxseed does not significantly affect total SFAs in the blood • Rumen-protected vs. Control 1000 Serum Total SFAs (ug/ml) 900 800 • P = 0. 07 700 • Rumen-protected vs. Ground flaxseed 600 500 • P = 0. 1 400 300 200 100 0 Control Ground Flaxseed Rumen-Protected Flaxseed

Rumen-protected flaxseed and ground flaxseed both successfully decrease SFAs in the milk 90 • Rumen-protected vs. Control 80 Milk SFAs (wt%) 70 • P < 0. 0001 60 • Rumen-protected vs. Ground flaxseed 50 40 • P = 0. 06 30 20 10 0 Control Ground Flaxseed Rumen-Protected Flaxseed

Rumen-protected flaxseed successfully raises omega-3 concentrations in the blood 800 • Rumen-protected vs. Control Serum Omega-3 (ug/ml) 700 • P = 0. 003 600 • Rumen-protected vs. Ground flaxseed 500 400 • P = 0. 01 300 200 100 0 Control Ground Flaxseed Rumen-Protected Flaxseed

Processing of flaxseed does protect PUFAs from biohydrogenation in the rumen • Rumen-protected vs. Control 3, 5 Milk Omega-3 (wt%) 3 • P = 0. 0004 2, 5 • Rumen-protected vs. Ground flaxseed 2 • P = 0. 007 1, 5 1 0, 5 0 Control Ground Flaxseed Rumen-Protected Flaxseed

We conclude this method of rumen-protection processing of flaxseed works • Increases the PUFA concentrations and lowers the SFA concentration significantly • Possible decrease in milk fat detected • Changes in milk fat and milk protein need more than 5 weeks in order to notice lasting effects (Oeffner et al. , 2013) • Further research is needed • Longer study with more animals • Supplementation Rate

Acknowledgments Seth Spencer, Farm Supervisor Russell Day and N 3 Feeds OSU Dairy Employees Volunteers The Cows

References Alvarado-Gilis, C. A. , C. C. Aperce, K. A. Miller, V. Bibber-Krueger, C. L, D. Klamfoth, and J. S. Drouillard. 2015. Protection of polyunsaturated fatty acids against ruminal biohydrogenation: Pilot experiments for three approaches. J Ani Sci 93: 3101– 3109. doi: 10. 2527/jas. 2014 -8015. Chen, S. , G. Bobe, S. Zimmerman, E. G. Hammond, C. M. Luhman, T. D. Boylston, A. E. Freeman, and D. C. Beitz. 2004. Physical and Sensory Properties of Dairy Products from Cows with Various Milk Fatty Acid Compositions. J. Agric. Food Chem. 52: 3422 -3428. doi: 10. 1021/jf 035193 z Drewnowski, A. 2011. The Contribution of Milk and Milk Products to Micronutrient Density and Affordability of the U. S. Diet. Journal of the American College of Nutrition 30: 422 S– 428 S. doi: 10. 1080/07315724. 2011. 10719986. Moallem, U. , D. Vyas, B. B. Teter, P. Delmonte, M. Zachut, and R. A. Erdman. 2012. Transfer rate of α-linolenic acid from abomasally infused flaxseed oil into milk fat and the effects on milk fatty acid composition in dairy cows. J Dairy Sci 95: 5276– 5284. doi: 10. 3168/jds. 2012 -5415. Polan, C. E. , J. J. Mc. Neill, and S. B. Tove. 1964. BIOHYDROGENATION OF UNSATURATED FATTY ACIDS BY RUMEN BACTERIA. J Bacteriol 88: 1056– 1064. Resende, T. L. , J. Kraft, K. J. Soder, A. B. D. Pereira, D. E. Woitschach, R. B. Reis, and A. F. Brito. 2015. Incremental amounts of ground flaxseed decrease milk yield but increase n-3 fatty acids and conjugated linoleic acids in dairy cows fed high-forage diets 1. Journal of Dairy Science 98: 4785– 4799. doi: 10. 3168/jds. 2014 -9115. USDA ERS - Dairy Data. Accessed February 26, 2017. https: //www. ers. usda. gov/data-products/dairy-data/.

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