SK281538B6 - Fodder additive to de-activate mycotoxins, process for producing thereof - Google Patents

Abstract

In the feed additive for the inactivation of mycotoxins in feed or in the digestive tract of animals, an enzyme preparation, made of microorganisms producing an enzyme from the group of bacteria, yeast or protozoa, is present in the feed, the enzymes produced by the microorganisms being selected from the group capable of forming epoxidases and / or lactonases. In use, the additive is preferably mixed with the feed and water at a pH of 1 to 4.5 at 15 ° C to 40 ° C for 3 to 30 minutes, and the mixture is then dried.

Description

Technical field

The present invention relates to a feed additive for the inactivation of mycotoxins in feeds or in the gastrointestinal tract of animals, wherein the feed comprises an enzyme preparation made from enzymes produced by organisms from the group of bacteria, yeasts or protozoa, using a feed additive for inactivating mycotoxins. the animal tract as well as the method of producing feed containing the feed additive.

BACKGROUND OF THE INVENTION

EP-A-0257996 discloses a feed premix as well as a process for its preparation which, in addition to a physiologically acceptable carrier, such as e.g. cereal flour, containing one or more enzymes. Also known from EP-A-0286056 is a compound feed as well as a process for processing a raw feed material which contains an enzyme preparation as an additive. Both known feed additives contain essentially amylase as an additive, since these grains which break down enzymes are lacking in young animals, especially pigs.

Further, for example, a feed additive is known from AT-PS 397756 which contains inorganic substances with a large inner surface, such as silicas or zeolites and organic heterocyclic polymers for the absorption of toxins.

Mycotoxins, which are poisonous products of fungal metabolism, are considered to be necessary in the animal and animal husbandry industry up to a certain degree and are known to a high degree as a cause of loss of production in the breeding and breeding of livestock. Therefore, various methods have been used for a long time to maintain the supply of mycotoxins to the animal's body to some extent, or to inactivate the animal's introduced mycotoxins in order to prevent the harmful action of mycotoxins in the animal's body. This mainly works because mycotoxins, based on the functional groups present in the molecule, usually show high chemical reactivity. This high chemical reactivity plays a central role in the toxicity ratios of such substances, as these groups, with biologically important molecules such as nucleic acids, enzymes, but also cell membrane lipids, perform a metabolism.

For the detoxification of mycotoxins, physical methods such as extraction, heating, irradiation as well as adsorption of mycotoxins, as well as their chemical inactivation, for example by treatment with acids, bases and various oxidizing agents, have been described so far. Of these various known methods, adsorption seems to be the most appropriate in practice. This adsorption is usually carried out by adding larger amounts of adsorbing agents such as clays, activated carbon or polymers. Of course, in this way, due to an efficient adsorption mechanism, it is by no means possible to fully inactivate or fix all mycotoxins to adsorbents. In essence, adsorption acts by forming a so-called hydration coating on the adsorption agent onto which the polar molecules are deposited on the basis of van der Waais forces. In this way, it has been possible to inactivate relatively mycotoxins such as aflatoxins to date in a relatively satisfactory manner. However, mycotoxins containing less strong polar functional groups can only be poorly bound to the adsorbent material in this way and sufficient inactivation of such mycotoxins is by no means possible by the prior art adsorption method.

VIS1 Atlas of Science: Biochemistry, Vol. 1, no. 4, 1988, Helmut Thomas et al., "Functions of epoxide hydrolases" describes detoxification with epoxide hydrolases of animal origin. On p. 288, it is reported that epoxide hydrolase activity is known to be present in all organisms. However, it may be classified only in higher life forms which have been shown to contain .. '

However, enzymes of animal origin cannot be considered comparable to those of microbial origin in function, since there is no link between the function of animal epoxide hydrolases and microbial origin.

Regardless of the specific activity of the enzymes, it is irrelevant that they can only convert substrates to which they have specific activity.

Also in the Chemical and Pharmaceutical Bulletin, Vol. 25, no. 12, 1977 Tokyo, p. 3410-3414, Yasuko Nakamura et al., "Reactivity of 12,13-epoxytrichothecenes with epoxide hydrolase, glutathione-S-transferase and glutathione" are degraded by aflatoxins using animal epoxide hydrolase.

Thus, the use of hydrobases of microbial origin for detoxifying mycotoxins is not described or apparent.

SUMMARY OF THE INVENTION

The invention is therefore directed to obtain a feed additive, in particular an enzyme preparation from enzymes produced by organisms from the groups of bacteria, yeasts or protozoa for the inactivation of mycotoxins in feedingstuffs or in the digestive tract, by which both mycotoxins usually present in feedingstuffs , so they could be widely removed.

The invention is further directed to the use of feed additives which could inactivate the mycotoxins present in the feed or in the digestive tract of the animals in order to prevent production losses in the production of livestock.

The solution to this object lies in the feed additive according to the invention in that the enzymes produced by the organisms are selected from the groups capable of forming epoxidases and / or lactonases. Since the enzymes produced by the microorganisms are selected from groups capable of producing epoxidases and / or lactonases, the mycotoxins contained in animal feed have been shown to exhibit high chemical reactivity and therefore are extremely strongly toxic, far-reaching enzyme preparations in the feed, possibly digestive tract of animals to degrade and / or convert them chemically to non-toxic or less toxic compounds and thus to significantly reduce their toxicity.

In particular, an enzyme preparation which is obtained advantageously by producing a pure culture or pure cultures of enzyme-producing organisms capable of producing epoxidases and / or lactonases, depending on their optimal growth conditions, is used herein. Thereafter, the cells of the cultures thus propagated are inactivated in a known manner to obtain enzymes which do not change in any way and thus remain intact. As an enzyme preparation, either these inactivated cells or the crude or pure extract from these cells can then be mixed directly into the feed. A particularly preferred method is the use of epoxidase and / or lactonase. Enzymes or enzyme preparations have the advantage that they can degrade or chemically convert mycotoxins, which generally fall into very different classes of chemical substances. This is particularly because many mycotoxins are cyclic hydrocarbon compounds with oxygen as the heteroatom, and the oxygen bond in these cycles is very often in the form of epoxides or lactones. The preferably used epoxidases, which belong to the group of hydrolase, are cleaved on the epoxide ring of the mycotoxin, thereby forming two hydroxyl groups, whereby the hydroxyl groups can subsequently be easily degraded in the animal body. Also preferably used lactonases act in an analogous manner. In order to achieve this effect, partial degradation of mycotoxins can already be achieved by feeding by mixing the feed, the feed additive according to the invention and the water, which is adjusted to a slightly acidic to neutral pH, and feeding it directly. Care must be taken that the feed slurry is not too strongly acidified, that the habit of feeding to animals or health problems is not adversely affected. Of particular importance in this context is that mycotoxin degradation with the enzyme preparation acts without additional energy and can only be achieved by homogeneously mixing the enzyme preparation into the feed in the environment of the digestive tract of the animals, or possibly already when mixed with water adjusted to an appropriate pH.

According to the present invention, an extract of Saccharomyces strains, in particular of Saccharomyces telluris, is particularly preferred as the enzyme preparation. Saccharomyces strains are special yeast species that are the most important groups in obtaining enzymes. Yeast can produce a wide variety of hydrolytic enzymes in extremely high amounts, and it has been shown that in the feed industry for quite different purposes, Saccharomyces strains can also be used for the particularly rapid and complete degradation of mycotoxins, which may contain both epoxide and lactone groups. In a particularly preferred method, Saccharomyces telluris is used with which the degradation of trichothesenes, in particular Τ-2-toxin, Η-Τ-2-toxin and deoxynivalenol, can be achieved under natural, digestive tract of animal conditions. In particular, the addition of Saccharomyces telluris could be a cleaved epoxy ring of said mycotoxins and the resulting dihydroxy compound will subsequently be metabolized in the gastrointestinal tract without demonstrating toxic efficacy.

In order to maximize the inactivation of the mycotoxins contained in the feed, the feed additive according to the invention is preferably formulated in addition to containing a natural or synthetic porous mineral with a larger inner surface in an amount of 0.5 to 8 kg / 1000 kg, especially 0.7 to 4 kg / 1000 kg feed. The feed additive of a mixture of enzyme preparation and natural or synthetic porous mineral with a large inner surface ensures that in an aqueous medium with a pH value corresponding to the digestive tract of the animals or in the digestive tract of the animals they adsorb to the mineral or are degraded directly by the enzyme preparation or chemically converted. Said natural or synthetic porous mineral with a large inner surface has the advantage that by adsorption on the surface of these minerals the mycotoxins are more readily available for further degradation by the enzyme preparation.

Kieselguhr, zeolites, 2- and 3-layer clays or mixtures thereof with a cumulative pore volume of from 0.05 to 4 cm ³ / g, in particular 0.5 to 2, are preferably used as natural or synthetic porous material with a large inner surface. 5 cm ³ / g and pH = surface area from 4.5 to 7.5. Diatomaceous earth, zeolites and / or layered clays, or mixtures thereof, having a cumulative pore volume of 0.05 to 4 cm ³ / g, in particular 0.5 to 4 cm ³ / g.

2.5 cm ³ / g, can particularly effectively adsorb mycotoxins and thereby inactivate to a large extent. Such minerals preferably have a pore diameter of from 10 nm to 1 µm and a surface pH of from 4.5 to 7.5. Such minerals are usually prepared in such a way that it is particularly advantageous to calculate the minerals at about 800 ° C to 900 ° C without the addition of flux, thereby achieving an even distribution of the pore surface structure. For example, mineral mixtures of 80% clinoptilolite and 20% muscovite, 80% chabasite and 20% muscovite, 75% clinoptilolite and 25% kaolinite as well as 80% chabasite and 15% kaolinite may be used.

Finally, the feed additive according to the invention for the inactivation of mycotoxins in feedingstuffs or in the digestive tract of animals is essentially characterized in that it consists of a mixture of 1 to 65% by weight, in particular 5 to 50% by weight, of enzyme preparation and 99 to 35% by weight. %, especially 95 to 500 wt. natural or synthetic porous minerals with a large inner surface. By using relatively large amounts of 1 to 65% by weight, in particular 5 to 50% by weight, of the enzyme preparation, it is ensured that the degradation of the mycotoxins present in the feed is achieved to a large extent. Furthermore, the abundance of natural or synthetic porous minerals with a large internal surface area ensures that those mycotoxins that are not available for chemical degradation by enzymes are essentially bound to the surface of these minerals and thus also no longer available for metabolism and can no longer adversely affect animals. In this connection, it has proven advantageous to adjust the activity of the enzyme preparation by admixing natural or synthetic porous materials for epoxidase at 470 to 600 units / g and for lactonase at 280 to 400 units / g.

The feed additives of the present invention with or without the addition of natural or synthetic porous minerals with or without a large internal surface can be particularly advantageously used for the inactivation or removal of aflatoxins, T-2 toxin, deoxynivalenol, nivalenol, zearalenone, monoacetoxyscirpen , HT-2 toxin, verrucarin, roridine, satratoxin, baccharinol, trichodermol, diacetoxyscirpenol, fumonism, ochratoxin, moniliformin, fusarenone X and crotocin in the feed or in the digestive tract of the animals.

In view of the wide range of mycotoxins that can be inactivated and / or degraded by the enzyme preparations of the present invention, the use of the feed additive of the present invention is a particularly advantageous and effective method for removing toxic substances from feed. The use of a feed additive ensures that also strongly mycotoxin-contaminated feeds can no longer be harmful to livestock and that in this way especially crops that have been subjected to a comminution, drying or especially processing to facilitate storage or feed mixtures after mixing with the feed additive according to the present of the invention without deliberation and without further examination by scientific institutes may be fed either directly or after treatment with water adjusted to a slightly acidic pH of the domestic animals.

The present invention further relates to a feed production process which is characterized in that the optionally ground and / or dried crops are mixed with the feed additive according to the invention as well as with water which is adjusted to a pH suitable for the reaction for 3 to 30 minutes. . at 15 ° C to 40 ° C and after adjusting to a pH suitable for feed and removal of liquid components, they are dried under the introduction of a stream of warm air. The pH adjustment or acidification can be carried out in this case with pharmaceutically acceptable acids, in particular hydrochloric acid3.

The pH is preferably adjusted to a value between 1 and 4.5 to achieve a corresponding rapid inactivation of the mycotoxins contained in the feed. According to the invention, the temperature range is set here between 15 ° C and 40 ° C, i.e.. j. essentially to room temperature to body temperature, for a period of from 3 to 30 minutes, to achieve further inactivation of mycotoxins. Since feed with a pH in the said strongly acidic range cannot be fed immediately by the animals, neutralization can be carried out after inactivation, for example by addition of sodium hydroxide, sodium bicarbonate or the like, to a slightly acidic to neutral pH. Further, after withdrawal of the liquid components, the feed is dried by supplying a stream of warm air to obtain mycotoxins free of long-term storable feed. Particularly preferably, the feed slurry is compressed into tablets before drying. Such tablets allow, due to the reduced surface, reduced oxidation of the feed nutrients while satisfying shelf life and transportability.

DETAILED DESCRIPTION OF THE INVENTION

The invention will be explained in more detail by means of the following examples.

Example 1

Study on the clinical effect of the feed additive of the invention on mycotoxins.

Feed additive:

% wt. % extract of strain Saccharomyces telluris containing epoxidases and lactonases; % kaolinite,% wt. % clinoptilolite.

48 hybrid pigs of both sexes with an average starting weight of 8.16 kg, a coefficient of variation of 20.83%, and an average age of 28.6 days were used for the experiment. Comparable piglets were selected for each stage of the experiment. Throughout the experiment, the temperature and humidity of the piglets were controlled.

The experimental groups were fed a diet containing three different mycotoxin contents and two different types of mycotoxins. In addition, four test groups received a feed additive according to the invention. One experimental group received neither mycotoxins nor the feed additive according to the invention.

The feed mainly contains soy, millet and milk powder and was administered 4 to 5 times a day, water was available from libitum. Mycotoxins were dissolved in ethanol, then mixed with a small amount of diet to be subsequently mixed into the feed. The groups formed are shown in Table 1, where the pigs of both groups with the same mycotoxins have the same age and the same origin.

Table 1

Experimental groups with different mycotoxin content with and without supplementation with feed additive

T2 * toxin + aflatoxin Bi feed additive 1 0.3 ppm + 0.1 ppm OkflÄ 2 0.5 ppm + 0.1 ppm Okha 3 δppm + 0. I ppm OkgA 4 0.3 ppm + 0.1 ppm 2 kg / t 5 0.5 ppm + 0.1 ppm 2kgA 6 1.0 ppm + 0.1 ppm 2knft 7 0.0 ppm -f * 0.0 ppm 2k »t 8 0.0 ppm + 0.0 ppm w

The parameters measured were body weight, food intake, average daily weight gain and rate of food conversion.

The results of the production parameters are shown in Table 2.

Table 2

Average food intake, daily average weight gain, food conversion rate of 8 experimental groups.

Group Average food intake Daily average oriraatok hmolnoati Transformation of food s 904 453 1.99 909 455 1.99 796 471 1.69 4 916 530 1.73 WITH 910 417 1.16 6 «II 492 1.66 922 520 * 1.77 WITH 199 509 1.77

*) Corrected by eliminating one death

Clinical evaluation:

Group 1 (0.3 ppm T-2 toxin + 0.1 ppm aflatoxin B1 and 0.0 kg feed additive):

Mild conjunctivitis, body condition and skin were normal, slightly reduced food intake was found.

Group 2 (0.5 ppm T-2 toxin + 0.1 ppm aflatoxin B1 and 0.0 kg feed additive):

Obvious and severe irritation of eyes, snouts and nose was observed, body condition was normal, appetite decreased.

Group 3 (1.0 ppm T-2 toxin + 0.1 ppm aflatoxin B ₍ and 0.0 kg feed additive):

All piglets show eye, nose and snout irritation and lateral throat and torax. All piglets show abdominal swelling, and the first recording of a decrease in food intake began at week 2. One piglet died on day 12.

Group 4 and 5 (0.3 ppm, respectively 0.5 ppm T-2 toxin + 0.1 ppm aflatoxin B] and 2 kg feed additive):

Light signs of conjunctivitis and skin irritation were observed, food intake and physical condition were assessed as normal.

Group 6 (1,0 ppm T-2 toxin + 0,1 ppm aflatoxin Bj and 2 kg feed additive):

Two piglets (out of 6) show only mild conjunctivitis, all other findings were in the normal range.

Group 7 (0.0 ppm, T-2 toxin + 0.0 ppm aflatoxin B] and 2 kg feed additive):

Group 8 (0.0 ppm T-2 toxin + 0.0 ppm aflatoxin B] and 0.0 kg feed additive) (comparison):

The piglets were healthy.

Analogous results were obtained using an enzyme preparation obtained from eubacteria.

From these evaluations, the positive effect of the feed additive of the invention on food intake is clear, weight gain and food conversion are evident at each of the three dose stages. Untreated groups exhibit much stronger clinical symptoms of mycotoxicoses than the groups treated with the feed additive. As in production parameters as well as clinically, there is a clear beneficial effect - in particular, more effective protection against T-2 toxin and aflatoxin contamination - of the feed additive of the invention.

Claims (9)

PATENT CLAIMS A feed additive for the inactivation of mycotoxins in animal feed or digestive tract based on an enzyme preparation made from enzymes producing microorganisms, from the group of bacteria, yeasts or protozoa, characterized in that the enzymes produced by microorganisms are selected from epoxidizable groups and / or lactonases. Feed additive according to claim 1, characterized in that the enzyme preparation is formed from a crude extract of an inactivated pure culture of enzyme producing organisms. Feed additive according to claim 1 or 2, characterized in that the enzyme preparation used is an extract of Saccharomyces strains, in particular of Saccharomyces telluris. The feed additive according to claim 1 or 2 or 3, characterized in that it additionally comprises a natural or synthetic porous mineral with a large inner surface. Feed additive according to claim 4, characterized in that diatomaceous earth, zeolites, 2- and 3-layer clays or mixtures thereof with a cumulative pore volume of 0.05 to 4 cm are used as natural or synthetic porous material with a large inner surface. ³ / g, especially 0.5 to 2.5 cm ³ / g and have a surface pH of 4.5 to 7.5. Feed additive according to claims 1 to 5, characterized in that it consists of a mixture of 1 to 65% by weight, in particular 5 to 50% by weight, of the enzyme preparation and 99 to 35% by weight, in particular 95 to 50% by weight. ., natural or synthetic porous minerals with a large inner surface. Use of a feed additive according to any one of claims 1 to 6 for the inactivation of aflatoxins, T-2 toxin, deoxynivalenol, nivalenol, zearalenone, monoacetoxyscirpenol, HT-2 toxin, verrucarin, roridine, satratoxin, baccharinol, fichodermcolol, trichodermcolol, diacetin moniliformin, fusarenone X and crotocin in the feed or in the digestive tract of the animals. Process for producing feed, characterized in that the optionally ground and / or dried crops are mixed with the feed additive according to any one of claims 1 to 6 as well as with water which is adjusted to a pH of 1 to 4.5 by addition of acid. , suitable for the reaction, is stirred for 3 to 30 minutes. at 15 ° C to 40 ° C and after adjusting the feed to a suitable pH and withdrawing the liquid components, it is dried by introducing a stream of warm air. A method according to claim 8, characterized in that the feed slurry is compressed into tablets before drying.