MXPA00009567A - Application of phytase in feed having low content of phytate - Google Patents

Abstract

The present invention relates to the use of the phytase enzyme in feeds having a low phytate content. The invention describes methods for feeding using a diet low in phytates supplemented with a phytase. Feeding an animal with a diet low in phytates comprises a substantial improvement of the animal's mineral state which results in improved zootechnical results with an improved growth rate and an improved feed conversion ratio. The invention also describes a feed, for animals, with low phytate content, which is supplemented with a phytase, as well as methods for the preparation of this feed.

Description

USE OF PHYTASE IN FEEDS THAT HAVE A LOW FITATE CONTENT Field of the Invention The present invention relates to the use of the phytase enzyme in feeds having a low phytate content.

Background of the Invention The diet of ruminants varies considerably in different domestic animal breeding systems, but also during different ages. In extensive animal husbandry systems, animals are typically fed hard fodder, such as grass, hay, grass silage and / or maize silage. Some feed or additional feed for livestock can be offered, as well as a vitamin mixture and / or similar. In these systems of raising domestic animals, young animals typically breast-feed with their mothers. In more intensive domestic animal husbandry systems, young animals, for example calves, are usually offered a milk substitute since the Ref: 1_.3391 Milk produced by cows is used for human consumption, or to process it into products for human consumption, such as butter and cheese. Calves intended for veal are usually offered primarily a milk substitute. Some other feed or livestock feed, such as hay, corn, or a special mixed concentrate can be offered additionally. Usually, these cattle foods require a low level of available iron, when the objective is to produce white beef. For calves that are intended for the production of red meat, feeds preferably contain some available iron. Older animals, such as dairy cows, cattle for meat, sheep, and deer, typically receive rations that contain hard fodder, for example, grass, hay, grass silage, and / or corn silage, livestock feed. concentrate, for example mixed concentrates (normally produced in a compound feed factory), cereals, mixed products and vegetable products and / or beer corujo. In addition, a mineral and / or vitamin mixture can be offered.

Phosphorus is an essential element for the growth of organisms. In livestock production, the feed must be supplemented with inorganic phosphorus in order to obtain a good growth performance in monogastric animals (for example, pigs, poultry and fish). Phytate occurs as a source of phosphorus storage, virtually all feed substances that originate from plants (for a review see: Phytic acid, chemistry and applications, E. Graf (ed), Pilatus Press, Mmeápolis, MN, USA (1986) Phytate comprises 1-3% of all nuts, cereals, legumes, seed oils, spores and pollen.Complete salts of citric acid are called citrines.Citrus acid is considered a factor anti-nutritional since it chews minerals such as calcium, zinc, magnesium, iron and can also react with proteins, and therefore, decreases the biodispomibility of the protein and of the nutritionally important minerals.The use of phytase in the nutrition of monogastric animals (eg pigs, coral birds and fish) has become widespread due to the availability of the microbial phytases at an affordable cost. The cloning and overexpression of microbial phytases results in a dramatic decrease in the price of the cost of the product which allows the commercialization of this enzyme for the nutrition of monogamous animals. In contrast, and typically, no inorganic phosphate is needed to be added to the livestock feed of old ruminants. It is believed that microorganisms present in the rumen produce sufficient quantities of enzymes that catalyze the conversion of the substrate (myo-inositol-hexate-phosphate) to mositol and inorganic phosphate. In Europe and the United States, livestock feed where phytase is advantageously applied, typically have a phytate phosphorus content of 0.5 to 0.35%. We now show that the addition of the phytase to the feed having a low phytate content such as the feed for young animals (ruminants or non-ruminants) results in a marked improvement in the daily gain and in the feed conversion ratio.

Description of the Figures Figure 1. Contents of zinc in the blood plasma as a function of time during the fattening period, for each of the four groups with a different composition of the diet: Group 1, ZnS0, and phytase; Group 2, only ZnSOa, - Group 3, only phytase; Group 4, without additions. Figure 2. Blood hemoglobin levels as a function of time during the fattening period, for each of the four groups with a difference and composition of diet: Group 1, ZnS0, and phytase; Group 2, only ZnS? 4, - Group 3, only phytase; Group 4, without additions.

Description of the Invention The present invention relates to the use of the enzyme phytase in the feed with a low content of phytate. In particular, the present invention relates to the use of the enzyme phytase in the feeding of animals that are typically fed diets low in phytate content, such as in poultry or young animals. According to one aspect of the invention, there is provided a method for feeding an animal wherein the animal is fed a diet with phytate supplemented with phytase. According to another aspect of the invention there is provided a method of feeding a young animal with a diet of phytate supplemented with phytase which is used as part of a method for reproducing and / or maintaining an animal.

No beneficial effect of phytase is expected when applied to feed with a low phytate content. Surprisingly, the present invention shows that the addition of exogenous phytase to low phytate feeds results in a marked improvement in zootechnical results, such as the average daily gain (ie, growth rate) and feed conversion, in animals fed with these low phytate feed containing phytase. Accordingly, an aspect of the invention provides a method for improving zootechnical results in an animal, wherein the method comprises feeding the animal with a feed with low phytate content supplemented with phytase. This improvement in the zootechnical result of animals fed a diet low in phytate supplemented with phytase may be apparent in any animal fed a diet ba in phytase In particular, an improvement in zootechnical results is quantified in young animals. The zinc content in the blood plasma and the hemoglobin levels also increase significantly as a consequence of the addition of the phytase to a feed with ba or phytate content while the levels of calcium, iron, magnesium and copper remain unaffected . The mere addition of a mineral such as zinc to the feed improves the mineral state of the animals, wherein the method comprises feeding the animal a diet low in phytate supplemented with phytase. In a preferred method according to the invention, the diet low in phytate with which the animal is fed also comprises zinc in addition to a phytase. Preferably, 0.005-0.2% (w / w) zinc is included in the diet, and more preferably 0.01-0.03% is included in the diet. The term "zootechnical results" is understood herein to mean animal performance in terms of daily gain (growth rate), feed conversion ratio (FCR), daily milk production and the like. The FCR is defined as the amount of feed (kg) that is required per kilogram of animal growth.

Through this invention, a feed or diet low in phytate 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 0.15%, and more preferably from 0.01 to 0.1%. More preferably, low phytate feeds have a phytate-P content of 0.01 to 0.05%, which is up to 10 times as high as in the feed of an older animal. Phytate can be determined as phytic acid, according to the method of for example, Oshima et al., 1964 (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.

For the purpose of this invention, young animals are defined as raised animals which are in their period of growth and / or fattening and / or which are still breastfed with their mother under natural conditions and / or receiving a specific food product. adapted for young animals, especially young ruminants, as defined hereafter. Young animals can be young ruminants such as calves, heifers, lambs, deer or goat calves. Animals young can also be non-ruminant young animals, such as piglets, accelerated broilers or young chickens. A phytase is defined herein as an enzyme which is a phosphatase capable of releasing at least one inorganic phosphate from a myo-inositol phosphate. A typical example of a phytase is a myo-inositol-hexate-phosphate-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 in any of the animal feedstuffs of the animal diet but rather is a supplement in cattle feed or in the diet in general. This means that the phytase that is applied in the methods and products of the invention requires any of the natural phytases that may be present in the feed for livestock, with respect to the amount of activity of the phytase and / or in the nature of the phytase, that is, the organism from which the phytase can be obtained and / or the structure and / or the biochemical qualities of the phytase molecule. Therefore, the phytase that is employed in the methods and products of the invention is a supplement to the feed with low phytate content of the animal. The phytase supplement can be an exogenously added phytase, which can be produced by fermenting host microbial cells expressing for the phytase. The cloning and overexpression of microbial phytases have been described in detail in EP-A-0 420 358. Another possibility of supplementing a feed with low phytate content and with phytase, is to add the material of the transgenic plant containing phytase , preferably by transgenic route, which has been genetically engineered 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 is a phytase with an optimum acid pH, that is, with an optimum pH less than 7.0, preferably less than 6.0. Preferably, the phytase can be obtained from plants or microorganisms. The microbial phytase can be obtained preferably from fungi, more preferably from fungi of the genus Aspergillus and Thermomyces. More preferably the microbial phytase can be obtained from black Aspergillus which belongs to the Aspergill us niger group as defined by Raper and Fenell (1965, In: The Genus Aspergillus, The Williams &Wilkins Company, Baltimore, pp. 234-344), such as Aspergillus niger, Aspergillus ficus and 5 Aspergillus awamori. In the methods and products according to the invention, preferably the phytase is added to the feed with low phytate content for an animal, in an amount that exceeds the amount of phytase activity that 10 is naturally present in any of the elements for livestock that are used ordinarily in the preparation of animal feeds. The level of activity of the phytase that is supplemented to the feed with low phytate content is preferably at least 15 10 FTU (for definition see Example 1) per kilogram of feed with a low phytate content, more preferably, at least 20 FTU per kilogram of feed with low phytate content, and more preferably at least 50 FTU per kilogram of feed with low phytate content, more preferably at least 100 FTU per kilogram low in phytate, more preferably at least 200 FTU per kilogram of feed with low phytate content. Normally the level of activity will be less than 10,000 FTU per kilogram of feed with ba or phytate content, preferably less than 5, 000 FTU per kilogram of feed with low phytate content, more preferably less than 2,500 FTU per kilogram of feed with ba or phytate content. According to another aspect of the invention, there is provided a method for preparing a low phytate feed containing phytase for the animals, wherein the livestock feed is mixed with phytase, and optionally with zinc. The phytase can be mixed with the livestock feed in dry form, for example, in the form of a granulated product containing an enzyme, or in liquid form, for example, of a stabilized liquid concentrate. In a preferred method the phytase is mixed with the livestock feed as part of a premix which may contain other food additives such as other enzymes, vitamins, or minerals. For animals that receive (mainly) a milk substitute, this time may be an optional way to offer the animal the phytase. A product according to the invention provides a feed with low phytate content comprising the components of the feed supplemented with phytase specifically adapted for this animal. In one embodiment of the invention, a feed with ba or phytate content can be provided by mixing the feed components with a high phytate content with the feed components with a low phytate content. In another embodiment of the invention, a feed with ba or phytate content can be provided by including a plant source in the feed which is genetically engineered and / or which is obtained by classical selection techniques to contain a lower amount of phytate of the amount that is present in the plant, unmodified and / or original. For example, the use of low-phytate corn or of soybean variants for the preparation of feed may provide feeds with a phytate-P content of about 0.2%. Another product according to the invention provides a feed with low phytate content specifically adapted for young animals, especially for young ruminants, supplemented with phytase. For example, a feed for calves destined for veal meat during the growing and fattening period that normally consists of mixtures of skimmed milk powder (and other dairy products) and dairy substitutes (milk substitutes) of vegetable origin such as soy isolates, soy and wheat concentrates as well as animal fat, vitamins and minerals. Some other feed or livestock feed such as hay, corn or a special mixed concentrate can be offered additionally. Therefore, a feed with low phytate content that is specifically adapted even in young animals, especially for young ruminants in its growing or fattening period is a substitute for milk that, in addition to the phytases, preferably comprise less than 60% (w / w of solid matter) of skimmed milk powder, more preferably less than 40%, more preferably less than 30%, and more preferably less than 20%, more preferably less than 10%, and most preferably still 0% . Preferably, the feed for young animals in the fattening or growing period comprises at least 5% milk substitutes of a vegetable origin, more preferably at least 25%, and more preferably at least 50%. A part of the dairy substitute produced is also used for monogastric animals, especially for piglets. Therefore, according to one aspect of the invention, there is provided a method for feeding an animal, such as poultry, with a low phytate feed supplemented with phytase. Preferably, this low phytates feed comprises a corn or soy source low in phytates which is genetically modified or obtained in a classical manner, and / or alternatively, another plant source low in phytates. According to another aspect of the invention there is provided a method for feeding a young animal during the period of growth and / or fattening, such as a young ruminant, with a diet low in phytate supplemented with phytase. The diet comprises dairy products, in which the amount of skimmed milk powder is preferably as low as possible, and dairy substitutes as described above that are supplemented with phytase. The invention is also directed to the non-therapeutic use of a phytase in any of the methods according to the invention such as for example a method of feeding an animal with a feed with low phytate content, the use of a phytase for the improvement of zootechnical results for the breeding and / or maintenance of this animal, as well as the use of a phytase to improve the condition of that animal. The following Examples serve to illustrate the invention and by no means are intended to limit the invention in any way.

EXAMPLE 1 Effects of microbial phytase on growth and feed conversion ratio up to 177 kg of live weight The tests are carried out with 4 groups of mottled steers. After a 6-week start period, the animals receive the following diets during a 20-week fattening period. The feed is supplemented with either microbial phytase (NATUPHOS f 5000, Gist-brocades, Delft, The Netherlands, which can be obtained from BASF, Lud ighshafen, Germany), ZnS04, or both. Group 4 serves as a negative witness. The microbial phytase is supplemented at a dose of 500 FTU (units of phytase) per kilogram of feed. The analytical method to determine the activity of the microbial phytase and the definition of a unit of phytase has been published by Engelen et al. (Journal of AOAC International 77 (3): 760-764 (1994).

Based on the composition of the diet in Table 1, the contents of various nutrients have been calculated as follows for all diets: Nutrient (g / kg): Crude Protein (185.2), Raw Fat (229.9), Raw Fiber (0.6), Humidity (35.9), Ash (62.3), Nitrogen free extract (451.1), Lactose (35.1), Starch (83.7), Protein derived from Dairy (51.8), Protein not derived from Milk (13.5), Lysine (17.9), Methionine (7.5), Met + Cys (10.4), Threonine (8.4), Tryptophan (2.2), Isoleucm (8.4), Calcium (8), Phosphorus (5.7), Fitato-P (0.5), Iron (23 mg / kg). ME = 4471.1 kcal / kg. Table 2 shows the average of food consumption, of growth and of the proportion of the feed conversion (FCR) that is calculated from the feed for each of the four groups.

Table 1. Composition of the diets for Groups 1-4 during the fattening period.

Table 2. Food intake, growth and feed conversion ratio of groups 1-4 during the fattening period.

The best results are obtained for Group 3 that is grown with the feed containing additional supplemental phytase (ZnS04).

Example 2 Effects of microbial phytase on zootechnical parameters up to 2067 kg live weight A similar experiment as described in Example 1 is carried out with larger animals. This is relevant for practice since the results obtained can vary as a function of body weight and age.

The study is carried out with calves destined for beef with a live weight of 130-150 kg at the beginning of the experimental period. In total, 24 black and white male calves are used for veal (Holstein-Friesian) for the experiment.

On day O all the animals are weighed and divided into three experimental groups in such a way that the groups are as homogeneous as possible. The allocation is based on the body weight and the hemoglobin content of the blood. The animals are housed individually under individual conditions of wooden boxes (75X175 cm) with floors made of slats. After the assignment, the animals are changed from a commercial diet of milk substitute to the experimental diet plus the premix. The composition of the diet is as described in Table 3. The microbial phytase is supplemented as NATUPHOS ® 5000 (Gist-brocades, Delft, The Netherlands, which is obtained from BASF, Ludwighshafen, Germany) in the diets.

During the experiment, the animals are fed according to the itinerary shown in Table 4. The animals are fed in the morning and in the afternoon. The use of milk that includes the phytase is dissolved in hot water (60-70 ° C), mixed for 3-5 minutes, followed by the addition of cold water while mixing until the The required amount of milk is prepared at a temperature of 40-41 ° C. Animals are not allowed to drink extra water.

Table 3: Composition of the diet for Groups 1-3 during the fattening period.

Fat composition: 37% bait, 40% shortening, 15% peanut fat, 5% lecithin, 3% emulsifier. The previous diet contains 0.04% phytate-P.

Table 4: Food program.

Animals are weighed before allocation and at 4, 8, 12 and 13 weeks after allocation with an accuracy of 0.2 kg. Food consumption is recorded by animal. With these figures and the results of the weight gain, the proportion of the feed conversion is calculated as kg of feed per kg of weight gain. Body weight (kg) at different times is shown in Table 5.

Table 5 Experimental groups receiving diets comprising the phytase show a daily gain in body weight substantially greater than the negative control group.

The cumulative proportion of the feed conversion ratio (kg of feed / kg of body weight gain) during the different periods for the three different groups is shown in Table 6.

Table 6 Experimental groups receiving diets containing supplemented microbial phytase show a considerable improvement in the feed conversion ratio.

EXAMPLE 3 Effects of microbial phytase on the general state Animals are grown as detailed in Example 1. Blood samples are taken at the beginning of the fattening period and at 2, 4, 8, 12, 16 and 20 weeks after . The blood is analyzed for the content of hemoglobin, iron, calcium, magnesium, copper and zinc, following methods known to persons skilled in the art. The contents of calcium, magnesium, copper and iron are the same for all experimental groups. The iron content in the blood plasma decreases too much for this period but does not differ significantly between the groups. There appear to be significant differences in the zinc contents in the blood plasma of the groups as shown in Figure 1. Group 1 receiving high levels of ZnSO4 and phytase in the diet shows the highest levels of Zinc in the blood plasma. Group 3, which receives only phytase and does not receive ZnS0, shows intermediate levels of zinc in the blood plasma. The same is true for Group 2 that receives supplemental zinc in the diet and does not receive phytase The Negative Control Group shows very low levels of zinc in the blood plasma. It is concluded that it is possible to improve ZnS04, by the addition of phytase or by a combination of both. The effects of age and composition of the diet on hemoglobin levels in the blood are shown in Figure 2. The implemented levels of hemoglobin in animals receive diets containing phytase and aggregated minerals (Group 3) is another indicator of a positive effect of the substance on the performance of the animals.

It is noted that in relation to this date, the best known method for the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects or products to which it refers.

Claims (19)

1. CLAIMS Having described the invention as above, property is claimed as contained in the following claims: 1. A method for feeding an animal with a feed supplemented with phytase, characterized in that this feed is a feed with a low phytate content that has a phytate-P content in a concentration ranging from 0.01 to 0.2% (w / w), preferably from 0.01 to 0.15%, more preferably from 0.01 to 0.1%, more preferably from 0.01 to 0.05%.
2. 2. A method for reproducing and / or maintaining an animal, characterized in that it comprises feeding the animal in accordance with the method of claim 1.
3. 3. A method for improving the zootechnical results of an animal, characterized in that it comprises feeding the animal in accordance with the method of claim 1.
4. 4. A method for improving the mineral state of an animal, characterized in that it comprises feeding the animal in accordance with the method of claim 1.
5. 5. A method according to any of the preceding claims, characterized in that the feed also comprises zinc.
6. 6. A method according to any of the preceding claims, of the phytate characterized in that the phytase is supplemented at an activity level of at least 10 FTU per kg of feed.
7. 7. A method according to any of the preceding claims, characterized in that the animal is a young animal.
8. 8. The method according to claim 7, characterized in that the young animal is a young ruminant selected from the group consisting of a calf, a heifer, a lamb, a young deer and a goat.
9. 9. The method according to claim 7, characterized in that the young animal is a piglet.
10. 10. The method according to claim 7, characterized in that the young animal is an accelerated broiler or a young broiler chicken.
11. 11. The method according to any of the preceding claims, characterized in that the feed is specifically adapted for a young animal.
12. 12. A method for preparing a feed for animals, characterized in that the method comprises the step of mixing the feed for livestock and a phytase.
13. 13. The method according to claim 12, characterized in that the phytase is mixed with the livestock feed as part of a premix.
14. 14. The method according to any of the preceding claims, characterized in that the Phytase can be obtained from a plant or a microorganism.
15. 15. The method according to any of the preceding claims, characterized in that the phytase can be obtained from a fungus, preferably from the fungus of the genus Aspergillus.
16. 16. An animal feed with low phytate content, which has a phytate-P content in a concentration ranging from 0.01 to 0.2% (w / w), preferably from 0.01 to O.lS ?, more preferably from 0.01 to 0.1% and more preferably from 0.01 to 0.05%, characterized in that it comprises the components of the feed specifically adapted for the animals and supplemented with phytase that does not occur naturally in the feed components.
17. 17. A feed according to claim 16, characterized in that the phytase is present at an activity level of at least 10 FTU per kg of feed.
18. 18. The use of a phytase for the improvement of zootechnical results in the reproduction and / or maintenance of animals.
19. 19. The use of a phytase for the improvement of the mineral state of the animals.