FR2714575A1 - New granules containing living organisms for animal feed. - Google Patents

Abstract

The present invention relates to novel granules having improved stability capable of being capable of subsequent granulation in an apparatus using moist heat and pressure as granulating means. These granules containing living matter are protected by gelatin and / or dextrin.

Description

NEW PELLETS CONTAINING ORGANISMS LIVING FOR
ANIMAL FEED
The present invention relates to novel granules containing a living material used in animal feed. It relates more particularly to new granules, having improved stability, containing either enzymes or bacteria and useful in animal nutrition. This stability can be improved with respect to the shelf life as well as with respect to external aggressions such as temperature and / or pressure.

 This invention is further directed to compositions containing living organisms which can be granulated by direct compression using heat and pressure without appreciable degradation.

 It is known from US Pat. No. 3,515,642 to prepare heat and pressure stabilized enzymatic compositions by adding to an aqueous solution containing an enzyme a sufficient amount of sorbitol to stabilize this enzyme, the aqueous solution being is then lyophilized and compressed. The composition thus obtained has improved wet heat stability when compared to the same composition not containing sorbitol, no test describes the preparation of granules by the joint action of heat and pressure and holding from the enzyme to such treatment.

 It is also known according to the patent published under number WO 92/12645 to prepare granules containing enzymes which can then be compressed to form, in the presence of moist heat, particulate foods for the animals.

 According to this patent the enzyme is protected, after deposition on a cellulose core, by a coating layer containing a mineral filler and a wax or a high melting point grease. This coated granulate can be subjected to compression temperatures, still more commonly known as "pelletization", reaching 70 ° C, the residual activity of the enzyme is still 90-100% calculated with respect to the initial activity. The invention described in patent WO 92/12645 has some disadvantages of implementation, it requires a series of successive coatings, the first is made with the enzyme, the second with wax or fat, the third with the mineral material and then started again with a layer of wax and a mineral material layer, this series of coating in onion film is long and expensive to implement.

 It has long been sought after as a cost-effective method of protecting living enzymes or bacteria to obtain pellets which could be pelletized without significant degradation of the enzymes. The present invention has made it possible to achieve this objective at a very reasonable cost and in a process quite different from those conventionally used in the field of enzyme protection.

The enzymes are marketed in liquid form or in powder form, often dried by lyophilization. When sold in liquid form, their stability at room temperature is often relatively short and their use is relatively difficult, because the amount biologically necessary to add to the food is very low, of the order of a few hundred milliliters for ton of finished product. Therefore, it is preferred to use dried enzymes which will then be mixed by successive steps with the food, the unprotected commercial enzymes have a double disadvantage, they are on the one hand powdery and secondly can not be mixed with feed and undergo direct pelletization without almost total degradation. The present invention has made it possible to prepare, intermediately, a granule containing the granular enzyme or bacterium to be protected, which can undergo a pelletization step without significant degradation. This granule also has a conservation over time and at room temperature quite amazing
The present invention relates to novel granules containing enzymes or living cells included in a gelatin or dextrin shell optionally containing an inert filler. Among the enzymes, there may be mentioned xylanases, cellulases, beta-glucanases obtained from Bacillus,
Actinomycetes, Fungi alpha amylases obtained from Bacillus, lipases, ligninases, phytases. These enzymes are used in livestock feed routinely, for example when poultry or pigs are fed with barley and / or oats, it is advisable to supplement this feed with food. α-amylase or 13-glucanase, when ruminants are fed with grain silage, it is advisable to add α-amylase or cellulases to their diet. Among the bacteria that can be added to animal feed are Lactobacillus, Pediococcus and
Enterococcus.

 The gelatin used in the present invention as a protective agent is used with or without crosslinking, ie it is simply dissolved in a necessary amount of water with the enzyme or bacteria and optionally an inert filler then the whole is dried. During this drying when it is desired to carry out a crosslinking, an optional spray of an aldehyde is carried out. The gelatin is preferably a gelatin having a bloom index of less than 50, this index is a mean index which can be obtained by mixing different gelatin.

It is obvious that gelatin with a higher bloom degree can also be used.

 The aldehyde used for the crosslinking of gelatin is chosen from, for example, acetaldehyde, glutaraldehyde and glyoxal. The crosslinking agent is used pure or in aqueous solution.

 Among the inert fillers include sugars such as glucose, lactose, celluloses, maltodextrins; glycerol, hydrophilic mineral fillers. All the elements of the composition are dissolved in aqueous solution and then dried in particular by any means which avoids carrying the enzyme or the bacterium above a temperature of approximately 500 ° C for a relatively long period of time (greater than about 15 minutes). Among the drying means include atomization, lyophilization. It is preferred to use atomization because it is a particularly economical drying means.

 Atomization is a technology that allows the passage of the previously described composition at a very high temperature but for a period so short that the enzyme or the bacterium undergoes almost no degradation during this type of drying.

 It is preferred to use an amount of enzyme calculated as weight of dried enzyme used relative to gelatin or dextrin between 5 and 50%. It is also preferred to use an amount of inert filler calculated by weight of filler relative to gelatin or dextrin of between 0 and 50 t. All of these solids are dissolved in a minimum amount of water of the order of about 100ml to 200ml per 100g of dry matter. The resulting liquid mixture is maintained at a temperature below 55 ° C, preferably between 40 and 55 ° C, and then introduced into an atomizer whose gas inlet temperature is between 100 and 300 ° C.

 The atomisate or lyophilizate obtained has a particle size of between 100 and 500 Cun and a bacterial population or an amount of enzyme of between 60 and 100% of the initial population included in the aqueous composition before atomization.

 The granule obtained is mixed with plant or mineral supports used for feeding the animals. Among the plant carriers that can be used, non-limiting examples include ground wheat flour, wheat remouldings, corn flour, ground soybean meal or soybean meal. The mixture of the granules containing the enzymes and / or the bacteria is granulated by passage through a die in which moist air is sent at a temperature between 65 and 90 C.

The preparation according to the invention has a significantly improved stability compared to all the solutions already recommended in the prior art.

 The present invention will be more fully described with the aid of the following examples which should not be considered as limiting the invention.

EXAMPLE 1
The coating solution is prepared by stirring in a 325 ml closed bottle of 30 g of 0 bloom gelatin, 15 g of lactose and 52 g of water brought to 60 ° C for 2 hours.

In a 250 ml beaker 26 g of bacteria are dissolved by stirring.
Enterococcus faecium, dose at 10.3 1010 cfu / g in 26 g of water at room temperature.

 The bacterial dispersion is then added to the coating solution. The temperature of the mixture drops towards 400C. According to the tests this temperature is maintained at 40 ° C with a water bath, is brought to 60 ° C in about 15 minutes.

The bacterial concentration of the starting suspension is 3.6% cfu / g.

This aqueous dispersion is then introduced into an atomizer
AEROMATIC, at the bottom of the tower by means of a nozzle. A protection and recovery filter is fitted at the top of the tank. The gas temperature at the inlet of the nozzle is 105 ° C, the air flow is 110-m3 / h, the residence time in the device is 20 minutes, the air temperature of output is 70-75 "C. The resistance results of the bacteria are shown in Table 1.

TABLE 1

<tb> Tests <SEP> Temperature <SEP> of <SEP> Duration <SEP> of <SEP> maintenance <SEP> Activity <SEP> Population
<tb><SEP><SEP> Blend <SEP> Blend <SEP> (h) <SEP> Ufc / g <SEP> Preserved <SEP> (%)
<tb><SEP> 1 <SEP> 40 "C <SEP> 0 <SEP> 32 <SEP> 109 <SEP> 88.9
<tb><SEP> 2 <SEP> 40 "C <SEP> 1 <SEP> 33 <SEP> 109 <SEP> 91.7
<tb><SEP> 3 <SEP> 40 "C <SEP> 2 <SEP> 32 <SEP> 109 <SEP> 88.9
<tb><SEP> 4 <SEP> 40 "C <SEP> 3 <SEP> 16 <SEP> 109 <SEP> 44.5
<tb><SEP> 5 <SEP> 60 "C <SEP> 0 <SEP> 79 <SEP> 108 <SEP> 21.9
<tb><SEP> 6 <SEP> 60 "C <SEP> 1 <SEP> 65 <SEP> 108 <SEP> 18
<tb><SEP> 7 <SEP> 60 "C <SEP> 2 <SEP> 30 <SEP> 108 <SEP> 8.3
<tb><SEP> 8 <SEP> 60 "C <SEP> 3 <SEP> 12 <SEP> 108 <SEP> 3,3
<Tb>
1 kg of atomisate obtained in test 1 are mixed with one ton of flour, analysis at time TO shows the presence of 106 cfu (colony-forming unit) / g, after 3 weeks the same analysis shows the presence of 10 6 cfu / g and 11 weeks after analysis showed the presence of 10 6 cfu / g.

EXAMPLE 2
Example 1 is reproduced by replacing weight for weight lactose with maltodextrin and bacteria with ss glucanase.

The dispersion is obtained by pouring the enzyme dispersion into the aqueous solution of the coating. Atomization is done in a device
AEROMATIC equipped with a 1.1 mm nozzle.

The activity of the atomized product corresponds to the theoretical activity of the starting product (2342 U / g), see results in Table 2 below.
TABLE 2

<tb> MALTODEXTRINE <SEP> YIELD <SEP> ACTIVITY <SEP> (U / G)
<tb><SEP> ATOMIZATION
<tb><SEP> Glucidex <SEP> 29 <SEP> 39.4 <SEP> 2320
<tb><SEP> Glucidex <SEP> 19 <SEP> 39.4 <SEP> 2550
<tb><SEP> Glucidex <SEP> 6 <SEP> 45.8 <SEP> 2380
<Tb>
EXAMPLE 3
According to the same procedure as in Example 1, a solution containing:
- glucanase 256 g
- lactose 148 g
- gelatin 0 bloom 296 g
- water 768g
This mixture is passed through a MINOR MOBILE atomizer with a nozzle of 2.1 mm in diameter and under a pressure of 1 bar. The air inlet temperature is 240 ° C, the outlet temperature is 110 to 1200C.

We collect at the exit:
1173 g of agglomerated particles having an average activity of 1740 units per gram corresponding to 74.3% of the theoretical activity of the engaged dry materials, the agglomerates are formed of a set of elementary beads whose diameter is greater than 25 μm .

 - 1174 gd, agglomerated particles having an average activity of 1420 units per gram corresponding to 60.6% of the theoretical activity of the engaged dry materials, the agglomerates are formed of a set of elementary beads whose diameter is greater than 50, um.

Some of these agglomerates are cross-linked with an aqueous solution of glutaraldehyde. The principle of crosslinking consists of dispersing the protected enzymes in a hot air stream while spraying a dilute solution of glutaraldehyde. This operation is done in an atomizer
AEROMATIC equipped with a 0.5 mm nozzle placed at the top of the tower, at a pressure between 0.4 and 0.5 bar. The injected drying air at 104 ° C has a flow rate between 50 and 100 m3 / h For 100 g atomisat, we use 3.12 g of pure glutaraldehyde which we dilute to 125 g with distilled water The injection time of the crosslinking solution is one hour followed by 5 minutes of drying.We recover more than 70 g of crosslinked protected enzymes in relation to the 100 g of protected enzymes introduced. show a color change with respect to the starting enzymes.

The protected enzymes are then mixed with a flour of the following percentage composition:
barley 50
but 17,37
corn gluten 3.00
soybean meal 15,75
meat meal 5.0
corn oil 6.0
carbonate of lime 0.45
dicalcium phosphate 0.43
wheat starch 1.0
growth premix 1.0
The composition of premix growth is as follows:
Vitamin Growth Complex 2.5
oligo chicken growth 5.0
choline 50GB 12.0
salt 30.0
calcium carbonate 49.5
antioxidant '1.0
For the manufacture of the flour we weigh and crush the cereals, we introduce into a mixer the ground cereals and minerals and the premix with the exception of wheat starch. The oil is sprayed under pressure using a nozzle, the flour is then put on hold.

 The protected enzyme, having undergone or not the crosslinking step, is assayed and then diluted a first time with a rewinding so as to obtain an enzymatic activity of about 200 Units AGL / g. The diluted enzyme is then mixed in a small mixer with the wheat starch not used in the manufacture of the flour, in these examples the 100 g of diluted enzyme are diluted again with 1 kg of wheat starch. This kilo is then added to 99 kg of previously prepared flour.

 Granulation is carried out in a horizontal KAHL press or in two vertical presses. The flour is introduced into the maturator, the steam is introduced into the maturator at a temperature and a rate such that the temperature measured on the flow of flour at the exit of the maturator is between 65 ° C and 95 ° C. The flour then falls on the die of the press and is compressed by the fluted rolls. After cutting the rods at the output of the die and measuring the temperature, the granules are cooled.

 The granules obtained are taken for analysis and stability over time.

The results are shown in Table 3.

EXAMPLE 4
Example 3 is repeated using the following mixture:
- glucanase 204 g
- Gelatin 140 blooms 356 g
- water 1040 g
The operating conditions are the same as in Example 3.

We collect at the exit:
850 g of agglomerated particle particles having an average activity of 1790 units per gram corresponding to 76.8% of the theoretical activity of the committed dried materials, the agglomerates are formed of a set of elementary beads whose diameter is greater than 25 fun.

 - 470 g of agglomerated particles having an average activity of 1470 units per gram corresponding to 63.1% of the theoretical activity of the committed dried materials, the agglomerates are formed of a set of elementary beads whose diameter is greater than 50 μm .

 These particles are granulated in the same way as those of the previous example, the survival results are shown in Table 3.

TABLE 3

<tb><SEP> NATURE <SEP> FROM <SEP> SAMPLE <SEP> TITLE <SEP> TITLE
<tb><SEP> flour <SEP> Yield <SEP> granulated <SEP> Rendemen
<tb><SEP> 20 <SEP> days <SEP> (%) <SEP><SEP> 50 <SEP> days
<tb><SEP> (u / kg) <SEP> (u / kg)
<tb><SEP> flour <SEP> single <SEP> no <SEP> granulate <SEP> 37 <SEP> 26
<tb><SEP> flour <SEP> granulated <SEP> to <SEP> 750C <SEP> 23 <SEP> 8
<tb><SEP> flour <SEP> granulated <SEP> to <SEP> 85 "C <SEP> 22 <SEP> 4
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 95 "C <SEP> 20 <SEP> 2
<tb><SEP> flour / enzyme / 0 <SEP> bloom <SEP> no <SEP> granulate <SEP> 224 <SEP> 215
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 75 "C <SEP> 108 <SEP> 45.4 <SEP> 85 <SEP> 40.7
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 85 "C <SEP> 41 <SEP> 10.2 <SEP> 28 <SEP> 12.7
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 95 "C <SEP> 22 <SEP> 1.0 <SEP> 10 <SEP> 4.2
<tb><SEP> flour / enzyme / 0 <SEP> bloom <SEP> tanned <SEP> 3 <SEP>% <SEP> 159 <SEP> 159
<tb><SEP> no <SEP> granulated
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 75 "C <SEP> 101 <SEP> 69.3 <SEP> 82 <SEP> 55.6
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 85 "C <SEP> 39 <SEP> 26.3 <SEP> 26 <SEP> 16.5
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 95 "C <SEP> 7 <SEP> 3.8 <SEP> 6 <SEP> 3.0
<tb><SEP> flour / enzyme / 140 <SEP> blooms <SEP> no <SEP> 175 <SEP> 175
<tb><SEP> granulated
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 75 C <SEP> 46 <SEP> 25.5 <SEP> 46 <SEP> 24.0
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 85 C <SEP> 18 <SEP> 9.4 <SEP> 16 <SEP> 7.6
<tb><SEP> flour <SEP> granulated <SEP> to <SEP> 95 C <SEP><10<SEP> 4 <SEP> 1,3
<tb> flour / enzyme / 140 <SEP> blooms <SEP> tanned <SEP> 3 <SEP>% <SEP> 167 <SEP> 177
<tb><SEP> no <SEP> granulated
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 75 C <SEP> 74 <SEP> 50 <SEP> 73 <SEP> 43.0
<tb><SEP> Flour <SEP> Granulated <SEP> to <SEP> 85 C <SEP> 53 <SEP> 40 <SEP> 29 <SEP> 16.6
<tb><SEP> flour <SEP> granulated <SEP> to <SEP> 95 C <SEP> 5 <SEP> 2,2 <SEP> 6 <SEP> 2,6
<Tb>

Claims (8)

 Granules having improved stability characterized in that they consist of a living material and gelatin and / or dextrin.  Granules according to claim 1 characterized in that they consist of a bacterium or an enzyme and gelatin and / or dextrin.  Granules according to Claim 1, characterized in that they consist of a bacterium or an enzyme, an inert filler and gelatin and / or dextrin.  Granules according to Claim 1, characterized in that the bacterium is chosen from Enterococcus, Pediococcus and Lactobacillus.  Granules according to Claim 1, characterized in that the enzyme is chosen from B glucanases and xylanases, cellulases, alpha amylases, lipases, lignignases and phytases.  Granules according to Claim 1, characterized in that the inert filler is chosen from sugars and glycerol.  - 7 - Process for the preparation of granules according to any one of the preceding claims, characterized in that an aqueous suspension is produced containing the bacterium and / or the enzyme, gelatin and optionally the inert filler which is then dehydrated .  - 8 - Process according to claim 7 characterized in that the dehydration is performed by lyophilization or atomization.