New feed ingredients based on glycerol esters

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Workshop on Glycerol Marketing, Uses and Chemistry Milano, 18/19 October 2012 New feed ingredients based on glycerol esters Manuela Parini Marketing director SILO S.p.A. - Firenze www.silospa.com

Role of lipids in the diets of food-producing animals Examples of amounts of supplementary lipids in the feeds Salmon: 10% Adult pigs: 1,5% Chickens: 5% Turkey: 7% Laying hens: 2% Energetic supplement Feed palatability Meat quality Eggs quality Energetic values of the vegetable oils that are commonly added to the feeds OIL AME Kcal/kg Broiler chickens > 21 days Crude Soybean oil 9,069 Crude Palm oil 7,978

Role of soybean & palm oil in farming animal diets Soybean oil oil is rich in linoleic acid (50-58%) and linolenic acid (5-10%) poli-unsaturated highly digestible fatty acids essential fatty acids (not physiologically synthetized by the animals) necessary to cell membranes, brain development is crucial in young animals diets Palm oil has a lower amount of linoleic acid (8-12%) and linolenic acid (traces) these fatty acids are easily oxidizable and they have to be reduced in the diets of: Adult pigs (Parma ham production) to avoid oxidation Adult chickens / turkey: to avoid oxidation in the meat and to prolong the shelf-life of processed meat Hens layers: to avoid eggs oversizing and eggs fragility is strategic in some growing periods

Palm oil sources available on the market OIL AME Kcal/kg Broiler chickens > 21 days Palm oil 7,978 PFAD (Palm distilled Fatty Acids) 6,678 Soybean oil = 9,069 Kcal/kg Glycerol can become an usefull tool for improving palm oil or palm fatty acids digestibility and cost / benefits ratio!!

Palm oil is less digestible than soybean oil because it is rich in palmitic acid placed in the position 2 of the triglyceride In the first intestinal tract lipases break the bonds fatty acid/glycerol in pos. 1 and 3. 2 free fatty acids and 1 monoglyceride with the fatty acid in pos. 2 are obtained. Monoglyceride Glycerol Palmitic Acid Oleic Acid Oleic Acid Free Fatty acids Lipases Pancreas

Free Palmit Acid, or Palmitic acid in position 1 & 3 are poorly absorbed In vegetable oils Palmitic acid is always in position 1 or 3 of the triglyceride. When it is separated from the glycerol by lipases, it is present in the gut as free Palmitic acid. Free Palmitic Acid tends to form indigestible soaps with the calcium, it is not absorbed but expelled with the faeces. MICELLES Oleic Acid Palmitic Acid Calcium Oleic Acid Bile Salts FAECES

ESTERIFICATION / RANDOMIZATION The recombination of free palm fatty acid with glycerol gives tow advantages: - Lower cost if compared with native palm oil - Higher digestibility duo to the shift of 33% of palmitic acid to position 2 of the triglycerides G L I C E R O L Oleic Acid Palmitic Acid Linoleic Acid

Palmitic acid in position 2 forms a monoglyceride If palmitic acid, by means of esterification, is shifted to the pos. 2 of the triglyceride, after lipases attack it remains bonded to glycerol as monoglyceride Monoglyceride of Palmitic Acid Oleic Acid Glycerol Palmitic Acid Oleic Acid Lipases Pancreas

Palmitic Acid Monoglyceride forms micelles and is absorbed MICELLES Palmitic Acid Oleic Acid Oleic Acid Palmitic Acid Monoglyceride is absorbed Bile Salts

Advantages of esterified oils Chickens in grower-finisher phase fed native palm oil or palm oil with palmitic and stearic acid in pos. 2 Digestion of Palmitic acid % Digestion of Stearic acid, % +40% +45% Adapted from W. Smink, Fatty acid digestion and deposition in broiler chickens fed diets containing either native or randomized palm oil. Poultry Science 87 - (2008).

ESTERIFICATION Another way to increase palmit acid digestibility is to add to the lipid mixture Monoglycerides produced through fatty acids forced glycerination G L I C E R O L Palmitic Acid G L I C E R O L Oleic Acid

Monoglycerides effects on palmitic acid digestion The absorption of free palmitic acid was measured in broiler chickens and compared with the absorption of palmitic acid mixed with free oleic acid, or with monoglycerides of oleic acid or with of monoglycerides of linoleic acid Lipid mixture in vivo absorption, % Palmitic acid Palmitic acid -3 Oleic acid + Palmitic acid (1) 11 1-Monoglycerides of Oleic acid + Palmitic acid (1) 28 2-Monoglycerides of Oleic acid + Palmitic acid (1) 24 1-Monoglycerides of Linoleic acid + Palmitic acid (1) 24 (1) Molar ratio of unsaturated lipids to palmitic acid : 0.8:1 (2) Mean ±SE of three individually caged cockerels with ligated pancreatic ducts. R.L. Garrett - R.J. Young Effect of Micelle Formation on the Absorption of Neutral Fat and Fatty Acids by the Chicken Journal of Nutrition - 1974.

Trial with piglets carried out by the Research Institute IRTA Barcelona March 2012 140 piglets from day 33 to day 47 of life were allotted in 3 groups and fed a commercial feed supplemented with 10% of native palm-oil or native soybean oil or esterified palm oil enriched with monoglycerides (MDT Palm) Lipid supplement in the feed (10%) Native Palm-oil MDT PALM Native soybean oil Average daily feed intake (grams/day) 435 409 407 Average daily weight gain(grams/day) 316 326 320 Feed Conversion Rate 1,38 ab 1,26 c 1,28 bc Live weight at 47 days of live (Kg.) 12,9 13,1 13

Advantages of esterified oils enriched with monoglycerides Native soybean oil vs esterified palmoil enriched with Monoglycerides 1 + 1 = 4

1 + 1 = 10 How can we achieve it? Glycerol BUTYRIC ACID BUTYRIC ACID BUTYRIC ACID Application Gastric-by-pass source of protected butyric acids for all monogastric species to be added to the feed; Beneficial effects gut & villi development, micro-flora modulation immuno-stimulation improvement of tight junction expression, strong protection against coccidiosis and enteric pathologies

Dr. Steve Leeson - Department of Animal & Poultry Science -Guelph, Ontario, Canada Trial with broiler chickens fed a diet supplemented with Butyric Acid Glycerides vs control diet Body weight and small Intestine characteristics of 21 days old male broiler chickens C4 Glycerides (*) Control Body weight (g) 992 a 862 b Small Intestine weight (SIW) (g) 24.7 a 19.0 b Small Intestine lenght (SIL) cm 163 a 142 b SIW / SIL g/cm 0.15 a 0.13 b (*) 6 kg of Liquid Butyric acid Glycerides 0-7 dd 3 kg of Liquid Butyric acidglycerides 8-20 dd (*) Butyric acid Glycerides produced & supplied by SILO SpA

University of Veterinary and Pharmaceutical Sciences Brno Institute of Pathological Morphology Prof. MVDr. Roman Halouzka DrSc. The effect of Butyric Acid Glycerides (*) on the height of intestinal villi in broiler chicken at day 14th of life Villi at the level of Meckel s diverticulum in Control group Villi at the level of Meckel s diverticulum in Butyric Acid Glycerides group (0,2% in drinking water) +35,8% (*) Butyric acid Glycerides produced & supplied by SILO SpA

Study carried out by Steve Leeson Guelph University 5 groups of broiler ROSS 308; 4 replicates per group (8 birds each replicate). All groups were vaccinated against coccidiosis at 1 st vaccine Vetech Immunocox; day of life with gel spray Supplements in the feed : - Control group : none - Sodium butyrate groups: 0,15% or 0,30% of protected sodium butyrate (0 14 dd) - Butyric acid glycerides (*) groups : 0,15% or 0,30% of Butyric acid glycerides (0 14 dd) At day 14 th of life an infection with th Eimeria Acervulina 5x10 5, Maxima 8x10 4, Tenella 1x10 5, Necatrix 6x10 4, was carried out. (*) Butyric acid Glycerides produced & supplied by SILO SpA

2 nd study carried out by S. Leeson Guelph University Coccidial infection at day 14 5 Groups of broiler chickens Body weight At 20 days/g Body weight gain 14-20 days /g (post challenge) % Mortality 14 20 days (post challenge) Control group 452 bc 84 25 a 0,15% Coated sodium butyrate 0,30% Coated sodium butyrate 0,15% Butyric acid Glycerides 455 bc 93 6,25 ab 450 c 112 25 a 498 ab 125 0b 0,30% Butyric acid Glycerides 512 a 136 + 52 gr!! 0b

Bacteria penetrate through intestinal lesions into blood stream Tight Junctions Butyric acid repairs intestinal lesions and blocks bacteria (barrier effect) Damaged Intestine Healthy Intestine

Energetic value of Butyric Acid Glycerides Trial carried out by Steve Leeson Guelph University - Canada FEED OXYGEN METABOLIC CAGE EXCRETA CO 2 128 one-day old broiler chicks were allotted to two groups: one control group and one experimental group. The feed of the experimental group was supplemented with 0,2% of butyric acid glycerides in replacement of 0,2% of corn. 8 replicated cages with 8 chicks each. Chicks were housed in an electrically heated brooder for first 18 day. At day 18, 3 chicks from each cage were moved to metabolic cages. Excreta were collected from day 9 to 12 and day 35 to 38 to estimate metabolizable energy. Growth performance was measured from day 1 to 18. Diets and excreta were analyzed for gross energy using an adiabatic, oxygen-bomb calorimeter and nitrogen using the Kjeldahl procedure (AOAC, 1990). Data were analyzed by t-test (SAS Institute, Inc. Cary, NC, USA).

Weight gain day 1 to day 18 (g) AMEn Kcal/kg Day 9 to day 12 Day 35 to day38 Control 630 3050 B 2977 0,2% Butyric Acid 647 3125 A 3025 Glycerides Δ 17 75 48 compared to 100% of Butyric acid glycerides 75 : 0,2 X 100 = 37,500 Kcal/ kg 48 : 0,2 X 100 = 24,000 Kcal/ kg Average = 30,000 Kcal/Kg 1 + 1 = 10

Conclusions Glycerol esterified with fatty acids gives the following advantages:.1. esterification of acid oils = more digestible oils due to the shift of 33% of saturated fatty acids to the position 2 of the triglyceride.2. forced glycerination = production of monoglyceides = higher absorption of saturated fatty acids.3. Esterification of butyric acid = unique tool for the animal gut development, pathogens control and higher nutrients absorption