ALKOSEL. The Pure Selenium Enriched Yeast Product The Bioavailable Selenomethionine. Lallemand Animal Nutrition

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ALKOSEL The Pure Selenium Enriched Yeast Product The Bioavailable Selenomethionine Lallemand Animal Nutrition

Selenium and Animal Nutrition ALKOSEL -TrialResults - Selenium s importance in animal nutrition - Biochemical role of selenium - Selenium, a varied soil distribution across the world - Different forms of selenium available -ALKOSEL : a monitored manufacturing process - Selenium bioavailability, an essential prerequisite -ALKOSEL : guaranted analytical characteristics -ALKOSEL : inclusion rates p3 p4 p5 p6 p9 p10 P13 p14 2

- Selenium s Importance in Animal Nutrition - Se Functions Anti-Oxidative Properties Immune Function Reproduction Function " A critical period : Calving even with high Vitamin E supplementation Main Problems linked with a Selenium + Vitamin E Deficiency! Immunity : Mastitis and somatic milk cell counts! Fertility : Placanta Retentions Uterin Infections Lower spermatic mobility! Oxidation : Myopathy : white muscle disease (NMD) Meat Quality (PSE) Red blood cell weakness : Haemolytic Anaemias (Lebreton et al., 1998) 3

- Biochemical Role of Selenium - # Glutathion Peroxidase (GSH-Px) : " Selenodependant enzyme, " Essential antioxidant : catalyses the conversion of hydrogen peroxide and organic peroxides to water and alcohols. # Selenium (GSH-Px) and Vitamin E : " Work synergetically to :! Maintain cells integrity,! Prevent from free radicals formation. " Vitamin E prevents from polyunsaturated fatty acids oxidation whereas Selenium (GSH-Px) metabolises peroxides that are already formed. 4

- Selenium, a Varied Soil Distribution across the World - Adequate Low Deficient No Data 5 (Oldfield, 1999)

- Different Forms of Selenium Available - # Inorganic Forms : Sodium Selenite (Na 2 Se0 3 )/Selenate (Na 2 Se0 4 ) " Low or no difference in the use of either selenite/selenate forms for specific selenoproteins synthesis, " Commodity Low differenciation Low price Almost systematic supplementation, " Contested bioavailability : selenium surplus is excreted through the urine rather than stored in the tissus. " High pulverulence causing inhalation dangers during workers handlings in factories, " Possible pro-oxidative oxidative properties of sodium selenite in vivo, that may result in an aditional need of antioxidants for the animals and that may lead to potential oxidation problem during (Spallholz, 1994) storage of premixes 6

- Different Forms of Selenium Available - # Organic Forms :ALKOSEL Mineral Enriched Yeast " Selenium as Selenomethionine, " Natural form encountered in crops and forages, " No pro-oxidative properties, " Higher bioavailability vs inorganic forms, " Mammals are not able to distinguish classic methionine form from selenomethionine, which is thus incorporated within body proteins, " Only available form of «protected» selenium. Selenium is an anion, thus it is not possible to obtain a «selenium chelate», as amino acids are also negatively loaded. 7

Alkosel - General Aspects - Supplementation Forms - Forms of selenium used to supplement foods and feeds are primarily the higher oxidation states of selenium while the selenium present in plants and of importance in nutrition contain selenium in the reduced state. Inorganic selenium is highly oxidised while the selenoaminoacids in plants and Alkosel are very reduced. The selenides are more available to ruminants because they avoid reduction to unavailable forms by rumen microbes. Availability of inorganic selenium is further depressed by low rumen ph on high concentrate diets. 8

- A Monitored Manufacturing Process - Inorganic Se Semet Analysis Spray dryer 9

- Selenium Bioavailability, an Essential Prerequisite - # Bioavailability Measured by : GSH-Px activity in the blood, (Pehrson et al., 1989) Se Concentration in the blood, (Malbe et al., 1995) Se Concentration in milk and tissus, (Pehrson et al., 1994) (Ortman, 1999) 10

Alkosel - General Aspects - Bioavailability - Inorganic selenium is passively absorbed from the small intestine while selenomethionine travels in the blood by amino acid transport mechanisms and can either be transportated to the liver for incorporation into selenoproteins or directly to other tissues for incorporation into tissue protein. Organic selenium is uniquely suited to metabolism because selenoamino acids can also be used for protein synthesis in muscle, mammary and other tissues. Tissue selenoamino acids will be recycled during cell turnover. Most of the inorganic selenium not used in selenoprotein formation is excreted in urine. 11

- A Higher Bioavailability of Selenium Yeasts - # In Ruminants : $ Part of selenite is transformed by rumen microbes into insoluble compounds which are only poorly absorbed through the intestinal tract. (Nicholson et al., 1991) $ Bioavailability of inorganic forms is particularly lowered in low ph ruminal conditions (high concentrate diets). 12

- Guaranteed Analytical Characteristics - # Guarantees : " Pure selenium enriched yeast product, " Total Selenium : 1900-2400 ppm, " Guaranteed Selenomethionine content : 2900-3500 ppm, analysed for each manufacturing batch, " Almost 70% of total selenium is from selenomethionine, " Nutritional content : B-group vitamins, amino acids, # Qualitative Analysis : " Total Selenium content, " Selenium Bound to the yeast (= Total Se Soluble Se), " Selenomethionine content. 13

- Inclusion Rates - " Maximum Se Inclusion Rate in EU : 0,5 ppm (total Se in finished feed) Concentration Inclusion 0,3 ppm Se Inclusion 0,4 ppm Se Inclusion 0,5 ppm Se Sodium Selenite 1% 10 000 ppm 30 g/ton 40 g/ton 50 g/ton ALKOSEL 2 000 ppm 150 g/ton 200 g/ton 250 g/ton 14

Selenium and Animal Nutrition ALKOSEL -TrialResults -ALKOSEL - Ruminants : Positioning " Positioning Dairy & Beef -ALKOSEL - Ruminants : Bioavailability " A higher bioavailability " A higher Selenium concentration in milk -ALKOSEL - Ruminants : Milk Quality/Health of Calves " A positive impact on milk quality and health status on calves " A positive impact on health status of calves " A positive impact on somatic milk cell counts P16 P16 P18 P18 P20 P21 P21 P23 P24 15

Alkosel - Positioning - Dairy -! Mastitis & Milk SCC : Herd selenium status is reflected in bulk tank SCC and incidence of subclinical mastitis. Selenium is needed for adequate activity of GSH-Px in phagocytic cells which fight intramammary infections. Infection of the mammary gland signals neutrophils and macrophages to leave the blood and enter the mammary tissue to locate the pathogens. Phagocytosis of the pathogen produce microbiocidal toxic O 2 metabolites. The toxic radicals are quickly neutralized by antioxidant enzymes including GSH-Px Px. An accumulation of toxins in the cell owing to selenium deficiency decreases killing ability. 16

Alkosel - Positioning - Dairy & Beef -! Reproduction : Reproduction selenium related problems such as retained placenta and other complications are associated with poor reproductive performance (CCI (IVV), service per conception, ). The aim is to improve herd selenium status,, allowing the cow to meet neonatal selenium needs without depleting her own reserves. The relationship of selenium to male fertility involves 3 different factors : antioxidant activity, sperm structure (reduced motility), testicular degeneration.! Improving Calf Selenium Status : Higher selenium status to meet requirements bring about selenium related problems : weak or stillborn calves, white muscle disease, e, lack of resistance to cold, stress at birth. 17

- A Higher Bioavailability - - Significative difference (p<0,05) in blood selenium concentration, - Larger difference in the milk (Alkosel: +130%), - Plateau reached after 1 week due to a possible substitution of methionine by selenomethionine within milk proteins. Number of animals : 42 dairy cows, production : 9000 L/hd Treatments : - Selenite : 0,24 0,31 ppm (DM) (Feb-May) - Selenate : 0,24 0,31 ppm (DM) - ALKOSEL : 0,24 0,31 ppm (DM) - Control : 0,10 0,12 ppm (DM) Homogeneization : No Se supplementation Sep-Feb (Se in feed = 0,10-0,12 ppm) Measures : - Blood samples, (Week 1,2 before suppl., week 1,2,3,4,6,8,10 et 12 after suppl.) - Milk samples. (same frequency) (Ortman et al., 1996) 18

- A Higher Bioavailability - GSH-Px activity significantively increased in all supplemented groups. The reduced trial duration can partly explain the fact that we did not reached a plateau. GSH-Px activity in erythrocytes permit to measure long-term Selenium status. Number of animals : 24 dairy heifers, Treatments : -I-Selenite: 0,25ppm(DM) (Feb-May) - II- Selenate : 0,25 ppm (DM) - III- ALKOSEL : 0,25 ppm (DM) - IV- Control : 0,03 ppm (DM) Homogeneization : No Se supplementation Sep-Feb (Se in feed = 0,03 ppm) Measures : - Blood samples, (Week 1,2 before suppl., week 1,2,3,4,6,8 et 11 after suppl.) (Ortman et al., 1996) 19

- An Improved Selenium Concentration in the Milk - Milk Se level is clearly superior in the Alkosel group and even for a low dose of 0,05 ppm. Aspila (1991) mentionned that milk for human consumption should at least contain 20 µg of Se per Kg of milk. Such a concentration is easily obtained by using an organic selenium source. Milk Se Concentration (in mcg/l) 35 30 25 20 15 10 0 50 119 163 273 Days Sodium Selenite ALKOSEL 0,20 ppm ALKOSEL 0,05 ppm Number of animals : 24 dairy cows, Treatments : Se in basic diet = 0,15 ppm (DM) (Nov-Sep) - Selenite : 0,20 ppm ie 0,35 ppm total - ALKOSEL : 0,20 ppm ie 0,35 ppm total - ALKOSEL : 0,05 ppm ie 0,20 ppm total Homogeneization : No Se supplementation Measures : - Blood samples, (d0, d50, d119, d163 and d273) - Milk samples (same frequency) (Pehrson., 1994) 20

- Milk Quality / Health of Calves - Control 10 DC Control 10 DC Control T Selenite ALKOSEL 17 DC 17 DC Selenite ALKOSEL 10 DC 7 DC 10 DC 7 DC Selenite Selenite ALKOSEL ALKOSEL S+ S++ A+ A++ No Se supplementation Se Supplementation Drying Period Calving 2 months after calving Number of animals : 44 dairy cows, Treatments : - Control : 0 Se (Nov-May) - Selenite : 0,20-0,23 ppm (DM) - ALKOSEL : 0,08-0,105 ppm (DM) For each treatment, DC are separated into 2 groups after calving Homogeneization : No Se supplementation Measures : - Blood samples, (calving, 2 months after calving) - Evaluation of health status (vigor, weight, myopathies, diahreas at birth, d20, d40) - Evaluation lactation parameters (production, mastitis) 21

- A Positive Impact on Somatic Milk Cell Counts - 50 % of DC with SCC >300000 40 30 20 10 0 T S+ S++ A+ A++ This trialshowedasignificantsignificant influence of selenium supplementation on somatic milk cell counts. Inthe Alkosel group during all the trial period, no animal had a somatic cell count >300 000. T S+ S++ A+ A++ + : Se supplementation from drying period to calving ++ : Se supplementation from drying to 2 months after calving 22

- A Positive Impact on Health Status of Calves - Frequency in % 90 80 70 60 50 40 30 20 10 0 T S+ S++ A+ A++ A Se supplementation markedly decreases risks of white muscle disease break out and gravity of subclinical muscular affections. Muscular Pbs White Muscle Disease + : Se supplementation from drying period to calving ++ : Se supplementation from drying to 2 months after calving 23

Somatic Milk Cell Counts/Milking (x10e+6) Alkosel - Ruminants - A Positive Impact on Somatic Milk Cell Counts - 3000 2500 2000 1500 1000 500 0 P<0,401 Control Alkosel Sem 0 Sem 8 P<0,013 This trial showed a significative influence of selenium supplementation on somatic milk cell counts. Corrected effects according to individual milk production (difference in milk production during the end of lactation phase). Number of animals : 100 mid lactating DC (calving March-April) Treatments : - Control : 0 Se (Oct-March) - ALKOSEL : 0,20 ppm (DM) Measures : - Milk and Blood Analyses, (week0, week1, week2, week4, week8, week14, week18, week26) - Milk Quality Parameters (neutrophil functions, somatic milk cell counts, bacteriology, ) (Malbe et al., 1995) 24

- A Positive Impact on Somatic Milk Cell Counts - Quarters harbouring positive bacteriology (%) 30 20 10 0 P<0,495 Control P<0,081 Alkosel At the beginning of the trial, 21% of quarters were infected. Staphylococcus aureus was isolated on 29% of positive quarters and Streptococci on 75% of cases. This trial showed a significative influence of selenium supplementation on bacteriology. Sem 0 Sem 8 Number of animals : 100 mid lactating DC (calving March-April) Treatments : - Control : 0 Se (Oct-March) - ALKOSEL : 0,20 ppm (DM) Measures : - Milk and Blood Analyses, (week0, week1, week2, week4, week8, week14, week18, week26) - Milk Quality Parameters (neutrophil functions, somatic milk cell counts, bacteriology, ) (Malbe et al., 1995) 25