1. AAFC-Lacombe

2. Why conduct beef fat research? A 1200 lb steer with ½ inch backfat, average muscling, yields a 750 pound carcass. The 750 pound carcass yields approximately: –490 pounds boneless trimmed beef – 150 pounds fat trim –110 pounds bone

3. Marbling fat is highly prized and is used to determine carcass value (grades A, AA, AAA, Choice). The value of trim fats is much lower than marbling and considered mostly a low vlaue waste.

4. Our beef fat research tries to change fat content and composition to add carcass value. Beef fat has been scorned by health officials due to saturated fatty acid (SFA) content, and its association with cardiovascular, diabetes and other diseases. We’re interested in investigating ways to improve the healthfulness of beef fat

5. In the past few years, the tide has shifted, and the healthfulness of lean beef is recognized to be on par with other protein sources such as chicken. The most consumed beef product is still, however, ground beef with 15-30% added trim fat. We’d like to convert the 150 lbs from low to highly valued healthy products.

6. Our interests are in parallel development of health and economic value To realize the potential, we have to pay attention along the development continuum….

7. Beef Composition

12. A more realistic way to improve the composition of beef fat might be by adding a PUFA source to the diet. Flaxseed’s a good choice due to it’s high content of omega-3 fatty acids (alpha-linolenic acid)? Research has been done on this in the past and the results were quite variable. We fed steers either 0 or 10% flaxseed in a barley grain based diet with 20% hay for 120 days.

13. Both ~4% total fat

14. When compared to a pig trial feeding 10% flax for 80 days…

15. Rumen Microbes Diet Rumen outflow Starch & Fibre Volatile fatty acids Low quality PTN & NPN (eg urea) Microbial PTN & ess. amino acids Polyunsaturated fatty acids (PUFA) Saturated fatty acids

16. PUFA Biohydrogenation = Detoxification Rumen bacteria SOAP Saturated Fat Firm Tallow Saturated Fat Firm Tallow

17. PUFA SFA 85-90% Efficient Our research is aiming to reduce conversion to SFA PUFA Hydrogenation

18. PUFA SFA 85-90% Efficient 1.We try to preserve PUFA 2.We also try to increase healthy intermediates like Rumenic Acid (CLA) and Vaccenic acid (t11-18:1) Anticarcinogenic improve blood lipid profiles anti-inflammatory properties PUFA Hydrogenation

19. Besides CLA and VA, there are at least 30 other BH products! PUFA SFA

20. We’re trying to figure out: – Production strategies to change BHP levels –Which of these BHP are “healthy” This info is needed to get BHP recognized by Health Canada and to establish amounts needed for things like health and enrichment claims

21. Feeding studies to increase hydrogenation Intermediates Rumen Microbiology Health Studies Obesity + Diabetes Cell culture: fat and cholesterol synthesis and inflammation Meat Quality Sensory Properties NIR for rapid analysis Genomic testing Hydrogenation Intermediates Methods for isolating Intermed. Drs. Dugan/Block/Baron Dr. Aalhus Drs. Prieto/Uttaro/Aalhus Drs. McAllister/Guan Prof. Guan Prof. Proctor Dr. Dugan Drs. Vahmani/Dugan/Meadus Anti-mutagenic activity (Ames Testing) Dr. Yang

23. Feeding Studies

24. –We’re still working on getting conditions right to produce high and consistent levels of ALA BH products. –But it’s a complex process with many interactions Fat/Oil Composition Amount Availabilit y Protein Fibre Starch Vit & Min Processing Bacterial Profile Growth Competitio n Rate of passage Meal Frequency Genetic controls Endogenous secretions Feed Intake Metabolism Rumination Fat synthesis

25. Rapid Analysis Comprehensive analysis of beef fatty acids is done using a combination of gas chromatography and high performance liquid chromatography. It takes about 6 hr of machine time plus time for sample and data processing. These methods are too slow to be used for quality control and sorting of high value carcasses and tissues.

26. Rapid Analysis We’ve been testing NIRS (near infrared-reflectance spectroscopy) to measure fatty acids in a few seconds. NIRS uses wavelengths in the NIR region (500-2500nm), and the pattern of wavelengths reflected from samples is used to predict composition. To date NIRS explains as much as 80% of the variation in sample BHPs, and as more samples are analyzed, we’re hoping for stronger preditions.

27. Rapid Analysis Another problem we’ve had to address is the large amount of between animal variation we see. If you’re feeding flaxseed to cattle for an extended period, it would be good to be able to sort out which ones deposit high levels of BHP. We found by measuring BHP in red blood cells we can accurately predict final tissue BHP within 2 months.

28. We now seeing if we can measure BHP in blood using NIRS. Vaccenic Acid % Vaccenic Acid

29. Rumen Microbiology Getting the rumen microbiology right will be a key to high and consistent levels of BHP in beef. Pathways for biohydrogenation have been worked out using bacteria isolated from the rumen But several other species are likely involved, haven’t been cultured and we don’t know how they interact. To tackle the problem we’ve taken advantage of advances in genomic technologies which have greatly increased the speed and reduce the cost of DNA sequencing.

30. Rumen Microbiology This involves pyrosequencing the 16s rRNA gene that has species-specific signature sequences. Using this, we were the first to demonstrate that the content of Vaccenic Acid in beef is correlated to 9 bacterial genera. We’re hoping to develop a rumen bacterial fingerprint to identify cattle that will deposit high levels of BHP, or develop a direct fed microbial to yield high and consistent levels of BHP in beef.

31. Health Studies A key to developing health and economic value of beef fat is to be able to increase the content of healthy BHP, and show that they have value when consumed in whole fats. This info is required to enable health and source claims. For work in this area we’ve collaborated with Prof. Spencer Proctor who is the Director of the Metabolic and Cardiovascular Diseases Laboratory at the U of A.

32. We fed BHP enriched beef fat to rats that develop obesity and diabetes. BHP enriched fat did not reduce blood triglycerides or cholesterol, but did reduce fasting plasma insulin. Further studies are planned to establish levels of BHP required for beneficial effects, and their mode of action. Health Studies

33. Isolating BHP To determine the effects of individual BHP, you have to be able to isolate and test them. We recently published a method to isolate several BHP from beef fat using a combination of silver-ion solid phase extraction and semi-preparative HPLC

34. Isolating BHP We’re also examining the potential to fractionate biohydrogenation products using molecular distillation

35. Cell Culture Studies When isolating small quantities of BHP, the most convenient way to test their effects is in cell culture. We’ve developed fat and liver cell culture models to do this. Our initial studies focused on trans fatty acids. Trans fatty acids have been in the news because they increase bad cholesterol and reduce good cholesterol in our blood.

36. Cell Culture Studies Most people studying trans fatty acids have used eladic acid (t9-18:1), which is the most concentrated in partially hydrogenated vegetable oil (i.e. margarine).. We’re interested in vaccenic acid (t11-18:1) and other trans fatty acids found in beef fat when feeding forages or flaxseed.

37. Cell Culture Studies We’ve found in cell culture the “ugly” trans turn on genes fat and cholesterol synthesis BUT other trans found when feeding flax (t11-, t13-, t14-, t15- and t16-18:1) don’t, and their effects are similar to oleic acid (major FA in olive oil). We’re continuing our work to characterize the effects of the trans fatty acids, and starting work with other BHP with more complex structures (i.e. dienes and trienes). RNA Micro -Array

38. Study Support: Peer Review Program

39. Collaborators Vern Baron Jon Meadus Hushton Block Dave Rolland Tim McAllister Spencer Proctor Jennifer Aalhus Bethany Uttaro Nuria Prieto Cletos Mapiye Tyler Turner Payam Vahmani The cast of many others from the the beef unit, abattoir and meat labs that do much of the hard work… Xianqin Yang Leluo Guan