Classifications

• • 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
  • 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
  • A23K20/00—Accessory food factors for animal feeding-stuffs
  • A23K20/20—Inorganic substances, e.g. oligoelements
  • A23K20/30—Oligoelements

Definitions

• the present invention relates to the application of the enzyme phytase in feed having a low content of phytate.
• the diets of ruminants vary considerably in different husbandry systems, but also at different ages.
• typically animals are fed with roughages, like grass, hay, grass silage and/or maize silage.
• Some additional feeds or feedstuffs may be offered, likewise a vitamin and/or mineral mixture.
• young animals In such husbandry systems, young animals typically suckle with their mother.
• young animals e.g. calves, are often offered a milkreplacer, since the produced milk of the cows is used for human consumption, or for processing to products for human consumption, like butter and cheese.
• Veal calves are usually mainly offered a milkreplacer.
• feeds or feedstuffs like hay, corn, or a special mixed concentrate may be offered additionally.
• these feedstuffs require a low level of available iron, when the aim is to produce white veal calf meat.
• the feeds preferably contain some available iron.
• Older animals like dairy cows, beef cattle, sheep and deer, typically receive rations containing roughages, e.g. grass, hay, grass silage and/or maize silage, and more concentrated feedstuffs, e.g. mixed concentrates (usually produced in a feed mill), cereals, - 2 -
• Phosphorus is an essential element for growth of all organisms. In livestock production, feed must be supplemented with inorganic phosphorus in order to obtain a good growth performance of monogastric animals (e.g. pigs, poultry and fish) .
• monogastric animals e.g. pigs, poultry and fish
• Phytate occurs as a storage phosphorus source in virtually all feed substances originating from plants (for a review see: Phytic acid, chemistry and applications, E. Graf (ed), Pilatus Press; Minneapolis, MN, USA (1 986). Phytate comprises 1 -3% of all nuts, cereals, legumes, oil seeds, spores and pollen. Complex salts of phytic acid are termed phytin. Phytic acid is considered to be an anti-nutritional factor since it chelates minerals such as calcium, zinc, magnesium, iron and may also react with proteins, thereby decreasing the bioavailability of protein and nutritionally important minerals.
• phytase in the nutrition of monogastric animals (e.g. pigs, poultry and fish) has become widespread thanks to the availability of microbial phytases at an affordable cost. Cloning and overexpression of microbial phytase has resulted in a dramatic decrease in cost price of the product enabling the commercialisation of this enzyme in the nutrition of monogastric animals.
• the feeds wherein phytase advantageously is applied typically have a phytate-phosphorus content of 0.25 to 0.35% .
• phytase to feed having a low phytate content such as feed for young animals (ruminants or - 3 -
• Figure 1 Contents of zinc in blood plasma as a function of time during the fattening period, for each of the four groups with a different diet composition: Group 1 , ZnS0 4 and phytase; Group 2, only ZnS0 4 ; Group 3, only phytase; Group 4, no additions.
• FIG. 1 Blood haemoglobin levels as a function of time during the fattening period, for each of the four groups with a different diet composition: Group 1 , ZnS0 4 and phytase; Group 2, only ZnS0 4 ; Group 3, only phytase; Group 4, no additions.
• the present invention relates to the application of the enzyme phytase in feed with a low phytate content.
• the present invention relates to the application of the enzyme phytase in the feeding of animals which typically are fed low-phytate diets, such as poultry or young animals.
• a method of feeding an animal wherein the animal is fed a low-phytate diet supplemented with a phytase.
• a method of feeding a young animal a low-phytate diet supplemented with a phytase which is applied as part of a method of breeding and/or keeping an animal.
• the present invention surprisingly shows that the addition of exogenous phytase to low-phytate feeds results in a marked improvement in zootechnical results, such as average daily gain (i.e. growth rate) and feed conversion, in animals fed such phytase-containing low-phytate feeds.
• one aspect of the invention provides a method of improving the zootechnical results of an animal, wherein the method comprises feeding the animal a low-phytate feed supplemented with a phytase.
• This improvement of the zootechnical results of animals fed with a low-phytate diet supplemented with a phytase may be apparent in any animal fed with a low-phytate diet.
• an improvement of the zootechnical results was measured in young animals.
• yet another aspect of the invention provides a method of improving the mineral status of animals, wherein the method comprises feeding the animal a low-phytate diet supplemented with a phytase.
• the low-phytate diet fed to the animal further comprises zinc in addition to a phytase.
• 0.005 - 0.2% (w/w) zinc is included in the diet, more preferably 0.01 - 0.03% is included in the diet.
• zootechnical results is herein understood to mean animal performance in terms of daily gain (growth rate), feed conversion ratio (FCR), daily milk production and the like.
• FCR is defined as the amount of feed (kg) required per kg of growth of the animal. - 5 -
• a low-phytate feed or diet is defined as a feed or diet comprising phytate-phosphorus (phytate-P) in a concentration ranging from 0.01 to 0.2% (w/w), preferably from 0.01 to 0.1 5%, and more preferably from 0.01 to 0.1 %.
• low-phytate feeds have a phytate-P content of 0.01 to 0.05%, which is up to ten times lower than the feed of an older animal.
• Phytate can be determined as phytic acid, according to the method of e.g. Oshima et al., 1 964 (Oshima, M., T.G.Taylor and A.Williams, Biochemical Journal 92, 42-46). Phytate-P is calculated as phytic acid * 0.29, since phytic acid contains approximately 29% phosphorus.
• young animals are defined as farmed animals which are in their growing and/or fattening period and/or which would still suckle with their mother in natural conditions and/or which receive a feed product specifically adapted for young animals, especially young ruminants, as defined hereinafter.
• Young animals can be young ruminants such as calves, heifers, lamb, deer calves or goats. Young animals can also be young non-ruminants, such as piglets, broilers or pullets.
• a phytase is herein defined as an enzyme which is a phosphatase capable of liberating at least one inorganic phosphate from a myo-inositol phosphate.
• a typical example of a phytase is a myo-inositol-hexakisphophate-3- phosphohydrolase (E.C. 3.1 .3.8).
• the phytase to be applied in the methods and products according to the invention is not present as a natural constituent of any of the feed stuffs in the animal diet but rather is a supplement to feed stuffs or the diet in general.
• the phytase applied in the methods and products of the invention differs from any of the natural phytases that might be present in the feed stuffs with respect to the amount of phytase activity and/or the nature of the phytase, i.e. the organism from which the phytase is obtainable and/or the structure and/or biochemical properties of the phytase molecule.
• the products of the invention is a supplement to the low-phytate feed of the animal.
• the phytase supplement can be an exogenously added phytase, which can be produced by fermentation of microbial host cells expressing the phytase. Cloning and overexpression of microbial phytases has been described in detail in EP-A-0 420 358.
• Another possibility of supplementing a low phytate feed with a phytase is to add phytase-containing transgenic plant material, preferable transgenic seed, which has been genetically engineered so as to express (or overexpress) a phytase as described in detail in EP-A-0 449 375.
• the phytase to be applied in the methods of the invention is preferably a phytase with an acidic pH optimum, i.e. with an optimum at a pH less than 7.0, preferably less than 6.0.
• the phytase is obtainable from plants or micro-organisms.
• the microbial phytase is preferably obtainable from a fungus, more preferably from a fungus of the genera Aspergillus and Thermomyces.
• the microbial phytase is obtainable from a black Aspergillus that belongs to the Aspergillus niger Group as defined by Raper and Fennell (1 965, In: The Genus Aspergillus, The Williams & Wilkins Company, Baltimore, pp 234-344), such as Aspergillus niger, Aspergillus ficuum and Aspergillus awamori.
• the phytase is preferably added to the low-phytate feed of the animal in an amount exceeding the amount of phytase activity naturally present in any of the feed stuffs ordinarily used in the preparation of animal feed.
• the activity level of the phytase supplemented to the low-phytate feed is preferably at least 1 0 FTU (for definition see Example 1 ) per kg of low-phytate feed, more preferably at least 20 FTU per kg of low-phytate feed, more preferably at least 50 FTU per kg of low-phytate feed, more preferably at least 1 00 FTU per kg of low-phytate feed, more preferably at least 200 FTU per kg of low-phytate feed.
• the activity level will be less than 10,000 FTU per kg of low-phytate feed, preferably less than 5000 FTU - 7 -
• a method for preparing a phytase-containing low-phytate feed for animals wherein feed stuffs are mixed with a phytase, and optionally zinc.
• the phytase may be mixed with the feed stuffs in dry form, e.g. as an enzyme containing granulate, or in liquid form, e.g. in the form of a stabilised liquid concentrate.
• the phytase is mixed with the feed stuffs as part of a premix which may contain other feed additives such as other enzymes, vitamins, minerals.
• this feed would be an optional way to offer phytase to the animal.
• a product according to the invention provides a low-phytate feed comprising feed components specifically adapted for the said animal supplemented with a phytase.
• a low-phytate feed may be provided by mixing feed components with a high phytate content with feed components with a low phytate content.
• a low-phytate feed may be provided by including a plant source in the feed which is genetically modified and/or obtained by classical selection techniques to contain a lower amount of phytate than the amount which is present in the unmodified and/or parental plant. For instance, the use of low-phytate corn or soy variants for feed preparation may provide feeds with a phytate-P content of about 0.2%.
• Another product according to the invention provides a low-phytate feed specifically adapted for young animals, especially young ruminants, supplemented with a phytase.
• a feed for veal calves in the growing and fattening period will usually consist of mixtures of skim milk powder (and other milk products) and milk substitutes (milk repiacers) of vegetable origin such as soybean isolates, soybean concentrates and wheat as well as animal fat and vitamins and minerals.
• feedstuffs like hay, corn, or a special mixed concentrate may be offered additionally.
• a low-phytate feed specifically adapted for young animals, especially young ruminants, in the growing and fattening period is a milk replacer which, in addition to phytase, preferably comprises less than 60% (w/w solid matter) skim milk powder, more preferably less than 40%, more preferably less than 30%, more preferably less than 20%, more preferably less than 1 0%, and most preferably 0%.
• the feed for young animals in the growing and fattening period comprises at least 5 % of milk substitutes of a vegetable origin, more preferably at least 25 %, most preferably at least 50% .
• Some milk replacer produced also is used for monogastric animals, especially for piglets.
• a method for feeding an animal such as poultry, a low-phytate feed supplemented with a phytase.
• said low-phytate feed comprises a genetically modified or classically obtained low-phytate corn or soy source, and/or alternatively, another low-phytate plant source.
• a method of feeding a young animal in the growing and/or fattening period such as a young ruminant, a low-phytate diet supplemented with a phytase.
• the diet comprises milk products, of which the amount of skim milk powder preferably is as low as possible, and milk substitutes as described hereinabove and is supplemented with a phytase.
• the invention is also directed to the non-therapeutic use of a phytase in any of the methods according to the invention such e.g.
• Trials are carried out with 4 groups of 1 9 piebald bull calves. After a starting period of 6 weeks animals received the following diets during a 20 week fattening period: feed was either supplemented with microbial phytase (NATUPHOS ® 5000, Gist-brocades, Delft, the Netherlands, obtainable from BASF, Ludwighshafen, Germany), ZnS0 4 or both. Group 4 served as a negative control. Microbial phytase was supplemented at a dose of 500 FTU (phytase units) per kg of feed. The analytical method for determining microbial phytase activity and the definition of a phytase unit has been published by Engelen et al. (Journal of AOAC International 77 (3) : 760-764 (1 994).
• Nutrient (g/kg): Crude protein (1 85.2), Crude Fat (229.9), Crude Fiber (0.6), Moisture (35.9), Ash (65.3), Nitrogen free extract (451 .1 ), Lactose (35.1 ), Starch (83.7), Dairy protein (51 .8), non-dairy protein (1 3.5), Lysine (1 7.9), Methionine (7.5), Met + Cys (1 0.4), Threonine (8.4), Tryptophan (2.2), Isoleucine (8.4), Calcium (8), Phosphorus (5.7), Phytate-P (0.5), Iron (23 mg/kg) . ME 4471 .1 kcal/kg.
• Table 2 shows the average feed intake, growth and feed conversion ratio (FCR) calculated therefrom for each of the four groups.
• Example 1 A similar experiment as described in Example 1 was conducted with older animals. This is relevant for practice since results obtained may vary as a function of body weight and age.
• the animals were housed individually under conventional conditions in wooden boxes (75 * 1 75 cm) with slatted floors.
• Microbial phytase was supplemented as NATUPHOS ® 5000 (Gist-brocades, Delft, the Netherlands, obtainable from BASF, Ludwigshafen, Germany) in the diets.
• mice were fed according to the schedule shown in Table 4. Animals were fed in the morning and in the afternoon. The milk replacer including the phytase was dissolved in hot water (60-70°C), mixed for 3-5 minutes, followed by addition of cold water while mixing until the required amount of milk with a temperature of 40-41 °C was prepared. The animals were not allowed to drink extra water. 12
• Fat composition 37% tallow, 40% lard, 15% coconut fat, 5% lecithin and 3% emulsifier.
• the above diet contained 0.04% phytate-P.
• Experimental groups receiving diets comprising phytase show a substantially higher daily gain in bodyweight than the negative control group.
• Example 1 Animals were grown as detailed in Example 1 . Blood samples were taken at the start of the fattening period and 2, 4, 8, 1 2, 1 6 and 20 weeks thereafter. Blood was analysed for the content of haemoglobin, iron, calcium, magnesium, copper, and zinc following methods known to people skilled in the art. Contents of calcium, magnesium, copper and iron were the same for all test groups. The content of iron in blood plasma decreased markedly during this period but did not differ significantly between the groups. There appeared to be a significant difference in zinc contents in blood plasma between the groups as shown in Figure 1 .
• Group 1 receiving elevated levels of ZnS0 4 and phytase in the diet showed highest levels of zinc in blood plasma.
• Group 3, receiving phytase only and no added ZnS0 showed intermediate zinc levels in blood plasma.
• the negative control group showed very low levels of zinc in blood plasma.

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• 1999
  • 1999-04-01 CN CN99804715A patent/CN1295443A/zh active Pending
  • 1999-04-01 CA CA002323581A patent/CA2323581A1/en not_active Abandoned
  • 1999-04-01 KR KR1020007010966A patent/KR20010042395A/ko not_active Application Discontinuation
  • 1999-04-01 AU AU38128/99A patent/AU748986B2/en not_active Ceased
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