WO2013174148A1 - Method for improving growth performance and/or bone performance of animal - Google Patents

Method for improving growth performance and/or bone performance of animal Download PDF

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Publication number
WO2013174148A1
WO2013174148A1 PCT/CN2013/000604 CN2013000604W WO2013174148A1 WO 2013174148 A1 WO2013174148 A1 WO 2013174148A1 CN 2013000604 W CN2013000604 W CN 2013000604W WO 2013174148 A1 WO2013174148 A1 WO 2013174148A1
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Prior art keywords
phytase
animal
ftu
per day
body weight
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PCT/CN2013/000604
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French (fr)
Inventor
Gilbert Weber
Jinlong WU
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Dsm Ip Assets B.V.
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Priority to AU2013265950A priority Critical patent/AU2013265950A1/en
Publication of WO2013174148A1 publication Critical patent/WO2013174148A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/03—Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03008—3-Phytase (3.1.3.8)
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/174—Vitamins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/189—Enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/20—Feeding-stuffs specially adapted for particular animals for horses
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
    • A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/03—Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03026—4-Phytase (3.1.3.26), i.e. 6-phytase
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/306—Foods, ingredients or supplements having a functional effect on health having an effect on bone mass, e.g. osteoporosis prevention

Definitions

  • the invention is related to a method for improving growth performance and/or bone performance of an animal.
  • the invention is related to a method for improving growth performance and/or bone performance of an animal by administering to the animal 25-Hydroxy Vitamin D 3 and phytase.
  • Phytase is a type of enzyme that catalyzes the hydrolysis of phytic acid, which is indigestible for organisms, to phosphorus which is digestible for organisms, including 3- phytase and 6-phytase.
  • Phytase has been widely used as animal fee additive, which may increase use efficiency of- phosphorus in plant feed by 60%, and decrease phosphorus excreted from feces by 40%, so it is significantly useful to improve production benefit in livestock, agriculture and reduce pollution of phosphorus from phytic acid, (see US 3,297.548; Nelson T. S. et al, J Nutrition, 101; 1289- 1294, 1971; and Nelson T. S. et at, Poult Sci. 47: 1842- 1848, 1968)
  • 25-Hydroxy Vitamin D has also been used widely as an animal feed additive.
  • US 2003/0170324 discloses a feed premix comprising 25-Hydroxy Vitamin D3 which may be added to poultry, swine, canine, or feline food
  • US 2005/0064018 discloses adding a combination of 25-Hydroxy Vitamin D 3 and vitamin D s to animal feed to improve swine 's bone strength
  • US 5,695,794 discloses adding a combination of 25- Hydroxy Vitamin D 3 and vitamin D 3 to poultry feed to ameliorate the effects of tibial dyschondroplasia.
  • the inventors of the invention found that a combination of 25-Hydroxy Vitamin D 3 and phytase can improve growth performance and/or bone performance of an animal and provide synergistic effect.
  • the invention provides a method for improving growth performance and/or bone performance of an animal, comprising administering to the animal an effective amount of 25-Hydroxy Vitamin D3 and phytase.
  • the term "animal” includes al l animals including human.
  • the examples of the animals include non-ruminants and ruminants. Ruminants include but are not limited to sheep, goat and cattle; and non-ruminants include but are not limited to horse; rabbit; pig including but not limited to infant pig, piglet, growing-fattening pig, sow and boar; and poultry such as turkey, duck and chicken (including but not limited to broiler chicken, egg-laying chicken) etc.
  • the term "growth performance” means average daily weight gain (ADWG), average daily feed intake (ADFi) and/or feed conversion rate (FCR) of the animal.
  • ADWG average daily weight gain
  • ADFi average daily feed intake
  • FCR feed conversion rate
  • bone performance means fresh weight, length, radius, breaking strength, and/or bone mineral density (BMD), and/or ash, Ca and/or P contents of the bone of the animal .
  • administration of 25-Hydroxy Vitamin and phytase to an animal can provide synergistic effects to improve growth performance and/or bone performance of the animal.
  • the term "synergistic effect” means the phenomenon of 1+1>2, i.e., two or more substances mutually provide better effect than the summation of effects produced by each one of the substances, or the phenomenon of "mutually promotive effect", i.e., the effect produced by one substance is reinforced in the presence of the other substances.
  • 25-Hydroxy Vitamin 3 ⁇ 4 may come from any source, including commercial routes or direct synthesis.
  • 25-Hydroxy Vitamin D ? of the invention is synthesized according to the method described in US 4,310,467 and US
  • the phytase may be any type of phosphatase enzyme that catalyzes the hydrolysis of phytic acid (myo-inositol hexakisphosphate) and releases a usable form of inorganic phosphorus.
  • the phytase of the invention may be 3-phytase, 6-phytase or a combination thereof.
  • the phytase of the invention may come from any source including plant and microbiological source, A preferable plant source for phytase is malt.
  • Preferred microbiological sources for phytase are Aspergillus oryzae, Aspergillus niger, Aspergillus jieuum and Peniophora lycii, and the phytase from Aspergillus niger has been commercialized for use in animal feed.
  • Phytase has also been isolated from E.coli, B. subtilis. and other thermostable microbes.
  • phytase Genes encoding phytase have been cloned from many microorganisms, and phytase can easily be produced and purified in industrial scale. Moreover, phytase can be genetically engineered such that it is more resistant to thermal, light, and/or chemical degradation. Examples of commercial phytase are Natuphos 5000 L, NatuphosTM 5000 G, Ronozyme® Hiphos M/L, etc.
  • a preferred phytase of the invention is 6-phytase (E.C, 3.1.3.26), preferably produced by a genetically modified strain Aspergillus oryzae (DSMZ 22-594).
  • FTU also-called FYT
  • FYT is the amount of phytase that liberates 1 pmole phosphate per minute at pH 5.5 and 37°C.
  • 25-Hydroxy Vitamin ⁇ % and phytase for administration depends on the particular conditions of the animal. Usually, based on the body weight of the animal to be administered, 25-Hydroxy Vitamin D3 is administered in an amount of 0,2-20 Mg/kg body- weight per day, preferably 0.5- 15 ⁇ /kg body weight per day, more preferably
  • the phytase is administered in an amount of 15- 150 FTU/kg body weight per day, preferably 20- 120 FTU/kg body weight per day, more preferably 30-100 FTU/kg body weight per day, even more preferably 50- 80 FTU/kg body weight per day, and most preferably 60 FTU/kg body weight per day.
  • the administration may be carried out once a day, or two or more times within one day to the animal.
  • 25-Hydroxy Vitamin D 3 and phytase may be administered simultaneously, or separately and successively, for example, administered in an interval of 10 mins. 20 mins, 30 mins, 40 mins, 50 mins, 1 h, 2 h, 3 h or 4 h.
  • the 25-Hydroxy Vitamin D 3 and phytase may be directly administered to the animal as a feed composition, or as components of an animal feed or in drinking water of the animal.
  • the invention provides use of a composition comprising 25-Hydroxy Vitamin D3 and phytase as an animal feed or feed additive.
  • the animal feed or the animal feed with addition of the feed additive comprises 5-500 ⁇ /kg, more preferably 10-300 .ug'kg, even more preferably 20-200 ⁇ ig/kg, even more preferably 40- 100 pg/kg, and most preferably 50 ug ' kg of 25-Hydroxy Vitamin D 3 ; and 500-4000 FTU/kg, more preferably 800-3000 FTU/kg, even more preferably! 000-2500 FTU/kg, even more preferably 1 200-2000 FTU/kg, and most preferably 1500 FTU/kg of phytase *
  • Piglets of an initial body weight of 7+0,5 kg were fed a basal diet ad libitum until 1O ⁇ 0.5 kg.
  • the formulation of the basal diet was as follows:
  • Premix provide in 1000 g of the basal diet: VA. 6600 IU; VD 3> 2000 IU; VH. 50 IU; VK 3) 2 mg; VB ; , 4.5 mg; VB 2 , 8 mg; VB 6 , 6 mg; VB i 2 . 60 ⁇ ; Niacin, 20 mg; pantothenicacid, 32 mg; Folic acid, 0.8 mg; Biotin, 0.23 mg; Choline, 750 mg; I ' e, 150 mg; Zn, 2340 mg; Cu. 120 mg; Mn, 50 mg; I, 0.6 mg; and Se, 0.3 mg
  • the 25-Hydroxy Vitamin D 3 used herein was obtained from DSM Nutritional Products Ltd.( Hy - DR. 1.25% Beadlet, Basel, Switzerland), and the 6-phytase used herein was also obtained from DSM Nutritional Products Ltd.(Ronozyme® Hiphos GT , Basel, Switzerland).
  • ADWG Average daily weight gain
  • ADFI average daily feed intake
  • FCR feed conversion rate
  • 25-Hydroxy Vitamin D 3 nor 6-phytase or with any one of 25-Hydroxy Vitamin l>, and 6- phytase the piglets administered with both 25-Hydroxy Vitamin D 3 and 6-phytase had quicker weight increase and lower FCR, so 25-Hydroxy Vitamin P 3 and 6-phytase had synergistic effect in improving growth performance of the piglets.
  • Example 2 Piglets of an initial body weight of 10 ⁇ 0.5 kg obtained from Example 1 were fed a basal diet ad libitum until 20 ⁇ 0.5 kg. Groups of 8 piglets each were fed as follows:
  • C basal diet + 1500 FTU/kg 6-phytase, and no 25-Hydroxy Vitamin D3 was added;
  • D basal diet + 50 .ug/kg 25-Hydroxy Vitamin D 3 + 1500 FTU/kg 6-phytase.
  • the formulation of the basal diet was as follows:
  • Premix provide in 1000 g of the basal diet: VA, 6600 IU; VD 3 , 2000 IU; VE, 50 IU; VK 3 , 2 mg; VBj, 45 mg; VB 2» 8 mg; VB 6 , 6 mg; VB 12 , 60 mi Niacin, 20 mg; pantothenicacid, 32 mg; Folic acid, 0.8 mg; Biotin, 0.23 mg; Choline, 750 mg; F ' e, 150 mg; Zn, 2340 mg; Cu, 120 mg; Mn, 50 mg: I, 0.6 mg; and Se, 0.3 mg
  • the used 25-Hydroxy Vitamin I> was obtained from DSM Nutritional Products Ltd.( Hy
  • ADWG Average daily weight gain
  • AD Ft average daily feed intake
  • FCR feed conversion rate
  • Femur fresh weight left femur was drawn out, muscles and hamstrings were stripped off to separate the femur (no injury of periosteum and cartilage cap), and then the femur fresh weight was measured.
  • Femur length was measured by Vernier Caliper after the femur fresh weight was measured.
  • Femur radius femur radius (cannon circumference of femur) was measured at a middle position after the femur fresh weight was measured.
  • Femur breaking strength was measured by Electronic Universal Material Mechanics Testing Machines (South China University of Technology, Guangzhou, China) to show the tibial strength. The span was set as 5 cm and the loading speed was set as 3 min/min. The bone was broken at a middle point. The sensor indicated the biggest force loaded during the breaking term, which was expressed as breaking strength (N).
  • Femur bone mineral density was measured by Bone Densitometer SD-200 (Beijing Search Institute of Nuclear Industry, Beijing, China) with a radioactive source of low energy nuclides (241 Am). The bone for measuring (fresh sample) was positioned under the detector, and the driving device was started to synchronously move the radioactive source and detector. The amount of radiation penetrating through the bone was detected and bone mineral content (BMC, g/cm) per unit of length was calculated based on the permeation rate. Femur bone mineral density (BMD, g/cm 2 ) was calculated based on the scan length and the bone mineral content (BMC).
  • Femur ash content After wrapped with filter papers, the femur was dipped in anhydrous ethanol for dehydration for 24 h and then extracted by ether for degrease for 24 h. After dried to constant weight at 100-105 °C, the dry weight of the dehydrated and degreased femur was recorded. Later, the femur was crushed, carbonized under low temperature until there was no smoke, transferred to a high temperature furnace for burning at 550 °C for 3 h, taken out into a drier for half an hour, and then weighed. It was put into the high temperature furnace for burning for 3 h, into a drier for cooling for half an hour, and then weighed again until the difference between two adjacent weighings was less than 1%. Finally, the ash content of the dehydrated and degreased femur was calculated by:
  • Ash Content (%) [ash content after ashed(g)J / [dry weight of degreased bone before ashed (g)] x 100% Femur ash
  • Ca content measured by EDTA direct volumetric method (see China State Standard GB/T 6436-92).
  • Femur ash P content measured by P-Mo-V acid colorimetric method (see China State
  • the data indicate that, the administration of 25-Hydroxy Vitamin D ; and 6-phytase improved bone performance of the piglets.
  • the simultaneous administration of 25-Hydroxy Vitamin D 3 and 6-phytase provided synergistic effects in improving bone mineral density and breaking strength, and increasing Ca and P contents of the bone.

Abstract

The invention provides a method for improving growth performance and/or bone performance of an animal. In particularly, the invention provides a method for improving growth performance and/or bone performance of an animal by administering to the animal 25-Hydroxy Vitamin D3 and phytase. The invention also provides use of a composition comprising 25-Hydroxy Vitamin DA and phytase as an animal feed or feed additive.

Description

METHOD FOR IMPROVING GROWTH PERFORMANCE AND/OR BONE PERFORMANCE OF ANIMAL
Technical Field
The invention is related to a method for improving growth performance and/or bone performance of an animal. In particularly, the invention is related to a method for improving growth performance and/or bone performance of an animal by administering to the animal 25-Hydroxy Vitamin D3 and phytase.
Background of Invention
Phytase is a type of enzyme that catalyzes the hydrolysis of phytic acid, which is indigestible for organisms, to phosphorus which is digestible for organisms, including 3- phytase and 6-phytase. Phytase has been widely used as animal fee additive, which may increase use efficiency of- phosphorus in plant feed by 60%, and decrease phosphorus excreted from feces by 40%, so it is significantly useful to improve production benefit in livestock, agriculture and reduce pollution of phosphorus from phytic acid, (see US 3,297.548; Nelson T. S. et al, J Nutrition, 101; 1289- 1294, 1971; and Nelson T. S. et at, Poult Sci. 47: 1842- 1848, 1968)
25-Hydroxy Vitamin D:> has also been used widely as an animal feed additive. For example, US 2003/0170324 discloses a feed premix comprising 25-Hydroxy Vitamin D3 which may be added to poultry, swine, canine, or feline food; US 2005/0064018 discloses adding a combination of 25-Hydroxy Vitamin D3 and vitamin D s to animal feed to improve swine 's bone strength; and US 5,695,794 discloses adding a combination of 25- Hydroxy Vitamin D3 and vitamin D3 to poultry feed to ameliorate the effects of tibial dyschondroplasia.
Surprisingly, the inventors of the invention found that a combination of 25-Hydroxy Vitamin D3 and phytase can improve growth performance and/or bone performance of an animal and provide synergistic effect.
Content of Invention
Therefore, the invention provides a method for improving growth performance and/or bone performance of an animal, comprising administering to the animal an effective amount of 25-Hydroxy Vitamin D3 and phytase.
In the invention, the term "animal" includes al l animals including human. The examples of the animals include non-ruminants and ruminants. Ruminants include but are not limited to sheep, goat and cattle; and non-ruminants include but are not limited to horse; rabbit; pig including but not limited to infant pig, piglet, growing-fattening pig, sow and boar; and poultry such as turkey, duck and chicken (including but not limited to broiler chicken, egg-laying chicken) etc.. In the invention, the term "growth performance" means average daily weight gain (ADWG), average daily feed intake (ADFi) and/or feed conversion rate (FCR) of the animal.
In the invention, the term "bone performance" means fresh weight, length, radius, breaking strength, and/or bone mineral density (BMD), and/or ash, Ca and/or P contents of the bone of the animal .
According to the method of the invention, administration of 25-Hydroxy Vitamin and phytase to an animal can provide synergistic effects to improve growth performance and/or bone performance of the animal. The term "synergistic effect" means the phenomenon of 1+1>2, i.e., two or more substances mutually provide better effect than the summation of effects produced by each one of the substances, or the phenomenon of "mutually promotive effect", i.e., the effect produced by one substance is reinforced in the presence of the other substances.
In the invention, 25-Hydroxy Vitamin ¾ may come from any source, including commercial routes or direct synthesis. For example, 25-Hydroxy Vitamin D? of the invention is synthesized according to the method described in US 4,310,467 and US
3,565,924.
In the invention, the phytase may be any type of phosphatase enzyme that catalyzes the hydrolysis of phytic acid (myo-inositol hexakisphosphate) and releases a usable form of inorganic phosphorus. The phytase of the invention may be 3-phytase, 6-phytase or a combination thereof.
The phytase of the invention may come from any source including plant and microbiological source, A preferable plant source for phytase is malt. Preferred microbiological sources for phytase are Aspergillus oryzae, Aspergillus niger, Aspergillus jieuum and Peniophora lycii, and the phytase from Aspergillus niger has been commercialized for use in animal feed. Phytase has also been isolated from E.coli, B. subtilis. and other thermostable microbes. Genes encoding phytase have been cloned from many microorganisms, and phytase can easily be produced and purified in industrial scale. Moreover, phytase can be genetically engineered such that it is more resistant to thermal, light, and/or chemical degradation. Examples of commercial phytase are Natuphos 5000 L, Natuphos™ 5000 G, Ronozyme® Hiphos M/L, etc.
A preferred phytase of the invention is 6-phytase (E.C, 3.1.3.26), preferably produced by a genetically modified strain Aspergillus oryzae (DSMZ 22-594).
The activity of phytase is measured in FTU units: 1 FTU (also-called FYT) is the amount of phytase that liberates 1 pmole phosphate per minute at pH 5.5 and 37°C.
The amount of 25-Hydroxy Vitamin Ό% and phytase for administration depends on the particular conditions of the animal. Usually, based on the body weight of the animal to be administered, 25-Hydroxy Vitamin D3 is administered in an amount of 0,2-20 Mg/kg body- weight per day, preferably 0.5- 15 μβ/kg body weight per day, more preferably
1-10 μgfkg body weight per day, even more preferably 1.5-5 μ§/な| body weight per day, and most preferably 2 μg/kg body weight per day; and the phytase is administered in an amount of 15- 150 FTU/kg body weight per day, preferably 20- 120 FTU/kg body weight per day, more preferably 30-100 FTU/kg body weight per day, even more preferably 50- 80 FTU/kg body weight per day, and most preferably 60 FTU/kg body weight per day.
The administration may be carried out once a day, or two or more times within one day to the animal. In addition, 25-Hydroxy Vitamin D3 and phytase may be administered simultaneously, or separately and successively, for example, administered in an interval of 10 mins. 20 mins, 30 mins, 40 mins, 50 mins, 1 h, 2 h, 3 h or 4 h.
The 25-Hydroxy Vitamin D3 and phytase may be directly administered to the animal as a feed composition, or as components of an animal feed or in drinking water of the animal.
Accordingly, in one embodiment, the invention provides use of a composition comprising 25-Hydroxy Vitamin D3 and phytase as an animal feed or feed additive.
Preferably, the animal feed or the animal feed with addition of the feed additive comprises 5-500 μΐ/kg, more preferably 10-300 .ug'kg, even more preferably 20-200 ^ig/kg, even more preferably 40- 100 pg/kg, and most preferably 50 ug'kg of 25-Hydroxy Vitamin D3; and 500-4000 FTU/kg, more preferably 800-3000 FTU/kg, even more preferably! 000-2500 FTU/kg, even more preferably 1 200-2000 FTU/kg, and most preferably 1500 FTU/kg of phytase *
The invention will be further described with reference to the following non-limiting examples.
Examples
Example 1
Piglets of an initial body weight of 7+0,5 kg were fed a basal diet ad libitum until 1O±0.5 kg. Groups of 8 piglets each were fed as follows:
A: basal diet, neither 25-Hydroxy Vitamin D3 nor 6-phytase was added;
B: basal diet + 50 ^ig/kg 25-Hydroxy Vitamin D¾ and no 6-phytase was added;
C: basal diet + 1500 FTU/kg 6-phytase, and no 25-Hydroxy Vitamin D3 was added;
D: basal diet + 50 μξ/kg 25-Hydroxy Vitamin D3 + 1500 FTU/kg 6-phytase.
The formulation of the basal diet was as follows:
Figure imgf000004_0001
Figure imgf000005_0002
* Premix provide in 1000 g of the basal diet: VA. 6600 IU; VD3> 2000 IU; VH. 50 IU; VK3) 2 mg; VB;, 4.5 mg; VB2, 8 mg; VB6 , 6 mg; VBi 2. 60 μ§; Niacin, 20 mg; pantothenicacid, 32 mg; Folic acid, 0.8 mg; Biotin, 0.23 mg; Choline, 750 mg; I'e, 150 mg; Zn, 2340 mg; Cu. 120 mg; Mn, 50 mg; I, 0.6 mg; and Se, 0.3 mg
The 25-Hydroxy Vitamin D3 used herein was obtained from DSM Nutritional Products Ltd.( Hy - DR. 1.25% Beadlet, Basel, Switzerland), and the 6-phytase used herein was also obtained from DSM Nutritional Products Ltd.(Ronozyme® Hiphos GT , Basel, Switzerland).
After the feeding term ended, the piglets were not fed for 12 hours and then weighed. Average daily weight gain (ADWG) during the term was calculated, average daily feed intake (ADFI) was calculated based on the feed weight before the feeding and rest feed weight: after the feeding, and feed conversion rate (FCR) was also calculated based on the average daily weight gain and average daily feed Intake (ADFI ADWG)). The experiment was run 6 times in parallel and averages were obtained as follows.
Figure imgf000005_0001
The data indicate that, compared with the piglets administered with neither
25-Hydroxy Vitamin D3 nor 6-phytase or with any one of 25-Hydroxy Vitamin l>, and 6- phytase, the piglets administered with both 25-Hydroxy Vitamin D3 and 6-phytase had quicker weight increase and lower FCR, so 25-Hydroxy Vitamin P3 and 6-phytase had synergistic effect in improving growth performance of the piglets.
Example 2 Piglets of an initial body weight of 10±0.5 kg obtained from Example 1 were fed a basal diet ad libitum until 20±0.5 kg. Groups of 8 piglets each were fed as follows:
A: basal diet, neither 25-Hydroxy Vitamin D3 nor 6-phytase was added;
B: basal diet + 50 .ug/kg 25-Hydroxy Vitamin D?, and no 6-phytase was added,
C : basal diet + 1500 FTU/kg 6-phytase, and no 25-Hydroxy Vitamin D3 was added; D : basal diet + 50 .ug/kg 25-Hydroxy Vitamin D3 + 1500 FTU/kg 6-phytase.
The formulation of the basal diet was as follows:
Figure imgf000007_0002
* Premix provide in 1000 g of the basal diet: VA, 6600 IU; VD3, 2000 IU; VE, 50 IU; VK3, 2 mg; VBj, 45 mg; VB2» 8 mg; VB6 , 6 mg; VB12, 60 mi Niacin, 20 mg; pantothenicacid, 32 mg; Folic acid, 0.8 mg; Biotin, 0.23 mg; Choline, 750 mg; F'e, 150 mg; Zn, 2340 mg; Cu, 120 mg; Mn, 50 mg: I, 0.6 mg; and Se, 0.3 mg
The used 25-Hydroxy Vitamin I>, was obtained from DSM Nutritional Products Ltd.( Hy
• DR. 1.25% Beadlet. Basel, Switzerland), and the used 6-phytase was also obtained from DSM Nutritional Products Ltd.(Ronozyme R Hiphos GT, Basel, Switzerland).
After the feeding term ended, the piglets were kept in limosis for 12 hours and then weighed. Average daily weight gain (ADWG) during the term was calculated, average daily feed intake (AD Ft) was calculated based on the feed weight before the feeding and rest feed weight after the feeding, and feed conversion rate (FCR) was also calculated based on the average daily weight gain and average daily feed intake (ADFI/ADWG). The experiment was run 6 times in parallel and averages were obtained as follows.
Figure imgf000007_0001
The above experimental data also indicate that the simultaneous administration of 25- Hydroxy Vitamin D:, and 6-phytase improved the weight gain and reduced FCR of the piglets and provided synergistic effects. Example 3
After the feeding of example 2 ended, fresh weight, length, radius, breaking strength, bone mineral density (BMD), and ash, Ca and P contents of the piglets' femurs were measured as below:
Femur fresh weight: left femur was drawn out, muscles and hamstrings were stripped off to separate the femur (no injury of periosteum and cartilage cap), and then the femur fresh weight was measured.
Femur length: Femur length was measured by Vernier Caliper after the femur fresh weight was measured.
Femur radius: femur radius (cannon circumference of femur) was measured at a middle position after the femur fresh weight was measured.
Femur breaking strength: femur breaking strength was measured by Electronic Universal Material Mechanics Testing Machines (South China University of Technology, Guangzhou, China) to show the tibial strength. The span was set as 5 cm and the loading speed was set as 3 min/min. The bone was broken at a middle point. The sensor indicated the biggest force loaded during the breaking term, which was expressed as breaking strength (N).
Femur bone mineral density: femur bone mineral density was measured by Bone Densitometer SD-200 (Beijing Search Institute of Nuclear Industry, Beijing, China) with a radioactive source of low energy nuclides (241 Am). The bone for measuring (fresh sample) was positioned under the detector, and the driving device was started to synchronously move the radioactive source and detector. The amount of radiation penetrating through the bone was detected and bone mineral content (BMC, g/cm) per unit of length was calculated based on the permeation rate. Femur bone mineral density (BMD, g/cm2) was calculated based on the scan length and the bone mineral content (BMC).
Femur ash content: After wrapped with filter papers, the femur was dipped in anhydrous ethanol for dehydration for 24 h and then extracted by ether for degrease for 24 h. After dried to constant weight at 100-105 °C, the dry weight of the dehydrated and degreased femur was recorded. Later, the femur was crushed, carbonized under low temperature until there was no smoke, transferred to a high temperature furnace for burning at 550 °C for 3 h, taken out into a drier for half an hour, and then weighed. It was put into the high temperature furnace for burning for 3 h, into a drier for cooling for half an hour, and then weighed again until the difference between two adjacent weighings was less than 1%. Finally, the ash content of the dehydrated and degreased femur was calculated by:
Ash Content (%) = [ash content after ashed(g)J / [dry weight of degreased bone before ashed (g)] x 100% Femur ash Ca content: measured by EDTA direct volumetric method (see China State Standard GB/T 6436-92).
Femur ash P content: measured by P-Mo-V acid colorimetric method (see China State
Standard GB/T6437-92).
Each group was measured twice and averages were obtained as follows:
Figure imgf000009_0001
The data indicate that, the administration of 25-Hydroxy Vitamin D ; and 6-phytase improved bone performance of the piglets. Special ly, the simultaneous administration of 25-Hydroxy Vitamin D3 and 6-phytase provided synergistic effects in improving bone mineral density and breaking strength, and increasing Ca and P contents of the bone.
The above examples illustrate the invention without limitation. It should be understood that any person skilled in the art may make obvious amendments and modification to the invention, which should be covered within the protection scope of the present invention.

Claims

Claims
1. A method for improving growth performance and/or bone performance of an animal, comprising administering to the animal an effective amount of 25-Hydroxy Vitamin D:, and phytase.
2. The method of claim 1, wherein the animal is selected from the group consisting of non-ruminants and ruminants, ruminants include but not limited to sheep, goat and cattle, and non-ruminants include but not limited to horse; rabbit; pig including but not limited to infant pig, piglet, growing-fattening pig, sow and boar; and poultry such as turkey, duck and chicken.
3. The method of claim 1, wherein the growth performance is average daily weight gain (ADWG), average daily feed intake (ADFI) and/or feed conversion rate (FCR) of the animal.
4. The method of claim 1, wherein the phytase is 3-phytase, 6 -phytase or a combination thereof.
5. The method of claim 4, wherein the 6-phytase is derived from a genetically modified strain Aspergillus oryzae (DSMZ 22594).
6. The method of any one of claims 1 to 5, wherein, based on the body weight of the animal to be administered, the 25-Hydroxy Vitamin Dj is administered in the amount of 0.2-20 tig/kg body weight per day, preferably 0.5- 15 μ§Λ§ body w eight per day, more preferably 1-10 pg/kg body weight per day, even more preferably 1.5-5 iig/kg body weight per day, and most preferably 2 pgfag body weight per day; and the phytase is administered in an amount of 15- 150 FTU/kg body weight per day, preferabl y 20-120 FTU/kg body weight per day, more preferably 30-100 FTU/kg body weight per day. even more preferably 50-80 FTU/kg body weight per day. and most preferably 60 FTU/kg body weight per day.
7. The method of any one of claims 1 to 5, wherein the 25-Hydroxy Vitamin I>, and the phytase are administered to the animal simultaneously.
8. The method of any one of claims 1 to 5. wherein the 25-Hydroxy Vitamin
Figure imgf000010_0001
the phytase are administered to the animal separately and successively.
9. The method of claim 8, wherein the 25-Hydroxy Vitamin D-, and the phytase are administered to the animal separately in an interval of 10 mins, 20 mins, 30 mins. 40 mins, 50 mins, 1 h, 2 h, 3 h or 4 h.
10. The method of any one of claims 1 to 9, wherein the 25-Hydroxy Vitamin D3 and the phytase are administered to the animal as a feed composition, or as components of an animal feed or in drinking water of the animal. feed or feed additive.
12. The use of claim 1 1 , wherein the phytase is 3-phytase, ό-phytase or a combination thereof.
13. The method of claim 12, wherein the 6-phytase is derived from a genetically modified strain Aspergillus oryzae (DSMZ 22594).
14. The use of any one of claims 11 to 13, wherein the animal feed or the animal feed with addition of the feed additive comprises 5-500 Hg/kg, preferably 10-300 fig/kg, more preferably 20-200 pg/kg, even more preferably 40-100 Hg/kg, and most preferably 50 μgίkg of 25-Hydroxy Vitamin D3; and 500-4000 FTU/kg, preferably 800-3000 FTU/kg, more preferablyTOOO-2500 FTU/kg, even more preferably 1200-2000 FfU/kg, and most preferably 1500 FTU/kg of phytase.
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WO2014102643A1 (en) * 2012-12-27 2014-07-03 Dsm Ip Assets B.V. Novel use of canthaxanthin and 25-hydroxy vitamin d3
WO2015091208A1 (en) * 2013-12-19 2015-06-25 Dsm Ip Assets B.V. Use of 25-hydroxy vitamin d3 to promote calcium absorption in ruminants
CN106719353A (en) * 2016-11-28 2017-05-31 平坝县鸿黔养殖场 A kind of efficient method for breeding of black goat
CN107821778A (en) * 2017-10-19 2018-03-23 界首市双马养殖专业合作社 A kind of meat sheep fattening period special feed
WO2020191426A1 (en) * 2019-03-22 2020-10-01 Jake Stockfeed Pty Ltd Horse supplement
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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014102643A1 (en) * 2012-12-27 2014-07-03 Dsm Ip Assets B.V. Novel use of canthaxanthin and 25-hydroxy vitamin d3
WO2015091208A1 (en) * 2013-12-19 2015-06-25 Dsm Ip Assets B.V. Use of 25-hydroxy vitamin d3 to promote calcium absorption in ruminants
CN106719353A (en) * 2016-11-28 2017-05-31 平坝县鸿黔养殖场 A kind of efficient method for breeding of black goat
CN107821778A (en) * 2017-10-19 2018-03-23 界首市双马养殖专业合作社 A kind of meat sheep fattening period special feed
WO2020191426A1 (en) * 2019-03-22 2020-10-01 Jake Stockfeed Pty Ltd Horse supplement
CN111955610A (en) * 2020-08-05 2020-11-20 中国农业大学 Application of 25-hydroxy vitamin D3 in improving pig skeleton quality

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