SK111694A3 - Fatty acid microspheres containing enteroccocus for use to enhance growth and improve meat quality - Google Patents

Abstract

Dried, rotary disc fatty acid microspheres of Enterococcus faecium, strains 301 and 202 are mixed and used as a feed additive for animals for growth enhancement and carcass quality improvement.

Description

Fatty acid microspheres containing enterococci, for use in promoting growth and improving meat quality

BACKGROUND OF THE INVENTION

Antibiotic growth promoters are widely used for poultry, especially chickens and turkeys. Growth enhancers such as Stafac and BMD (bacitracin methylenedisalicylate) are known antibiotics and are used in subtherapeutic doses, for example 10 g / ΐ and 25 g / t, as feed additives to promote the desired growth characteristics in poultry. However, the use of antibiotics for this purpose has recently been the subject of some criticism. One objection is that poultry will develop tolerance to antibiotics over time, and eventually the antibiotic will no longer favor growth promotion. Other objections concern the health risks of unnatural antibiotic additives and the worsening effects they may have. However, because of their advantages, antibiotics have been commonly used to improve feed conversion, improve meat composition, and promote growth.

It is known that certain bacteria, when added to feed, have a potentially beneficial effect. This beneficial effect consists in the delivery of natural intestinal microflora. Some companies offer for sale directly fed microbial products containing the desired bacteria. Directly fed microbial compositions, however, create some difficulties in maintaining product stability. Typically, the directly fed microbial composition is added to the feed in a relatively low amount, for example about 0.1%. However, unused feed or feed additive containing directly fed microbial preparation is often stored on farms for a long time. This storage often takes place under conditions in which there is a certain humidity and high temperature. In many cases, just enough moisture is present to activate the start of bacterial growth, but not enough to support it. As a result, the bacteria die and the activity of the directly fed microbial preparation disappears. In other cases, the antibiotics added to the feed or feed additive containing the directly fed microbial preparation adversely interact with the bacteria, especially when a small amount of moisture is present, and the bacteria die again. Thus, there is a significant problem of long-term storage stability of directly fed microbial formulations.

In another environment, when the directly fed microbial composition is added to, for example, chicken feed, it is usual to pellet the material and add the directly fed microbial composition prior to pelletization. Moisture from steam used in. pellets, partially activating the bacteria, but may cause their disappearance if moisture is insufficient. Bacteria can also die due to heat during pelletization. Then there is also the problem of the acidic environment in the stomach, the potentially inactivating bacteria, before they actually enter the intestines. Thus, there is a continuing need for directly fed microbial formulations that release microorganisms only at the right time in the intestine and not more due to moisture or unfavorable pH conditions such as exist in the digestive tract before the small intestine.

In poultry it is particularly desirable to achieve certain characteristics as far as possible. These include increased weight gain, better feed conversion, meat composition, and finally a uniform flock weight. Increased weight gain and better feed conversion are, of course, desirable for the related savings that accompany these desirable results. The composition of the meat is important because the most suitable area for tissue storage in order to obtain select meat is the chest. Thus, not only the weight gain is important, but also the location where it is stored. The uniform weight of the flock is important because if more birds are of normal size, less handwork is needed and the processor can rely more on machine processing. On the other hand, if birds vary widely in size from very small to very large, while maintaining the total weight of the flock, smaller and larger birds require much more manual work and cannot be easily processed by the machine due to their non-uniform size. A uniform flock size with a high percentage of distribution within the normal size range allowing processing of the chickens by a standardized device is therefore a desirable characteristic.

Similarly, a microbial composition directly fed not only to poultry such as chickens and turkeys, but also suitable for pigs would be of great benefit.

The primary purpose of the invention is to find a directly fed, antibiotic-free poultry microbial composition containing only fatty acid microspheres containing naturally occurring organisms.

Another primary purpose of the invention is to find a directly fed microbial composition comprising two organisms, namely Enterococcus faecium 301, no. DSM DSM Nr. And Enterococcus faecium 202, no. DSM DSM Nr. 4788. DSM is an abbreviation of the German collection of bacterial cultures of Deutsche Sammlung von Mikroorganismen, located in Braunschweig, Germany. These organisms will be deposited in the ATCC with the release of all restrictions following the ADR report.

Another object of the invention is a directly fed microbial composition which provides an increased weight gain in poultry, which provides better feed conversion, which provides a higher yield of pork, and which provides a uniform flock weight within the normal size range.

Another primary purpose of the invention is a directly fed microbial composition suitable as a feed additive for poultry containing microsphere-containing bacteria using a special rotation technique using a free fatty acid matrix.

Another object of the invention is a directly fed microbial composition that is stable for a period of 3 to 6 months without significantly reducing the number of organisms.

baker i i

Another object of the invention is a rotary molding method which makes it possible to obtain a unit of dried size.

Another object of the invention which are rotary discs, batches for poultry, are free-flowing, dry-powdered bacteria and easily processed into feed

It is further an object of the present invention to provide them with feeds as directly fed and pigs.

is a microsphere wherein the microbial fatty acid these beads are suitable for poultry compositions

SUMMARY OF THE INVENTION

SUMMARY OF THE INVENTION The present invention provides a method and composition for promoting the growth of poultry and pigs, comprising adding a small but effective amount of a directly fed microbial composition containing dried Enterococcus faecium 301 microspheres, No. DSM to a conventional feed ration.

DSM-Nr. 4789, fatty acid-based and dried microspheres Enterococcus faecium 202, No. DSM DSM-Nr. 4788, fatty acid-based, wherein the microspheres are preferably formed by rotary disk drying.

Surprisingly, it has been found that the growth promotion of poultry and pigs can be achieved by adding a certain amount of Enterococcus faecium 301 microspheres to conventional feed rations. DSM DSM Nr. 4789, fatty acid-based, and some microspheres of Enterococcus faecium 202, no. DSM DSM Nr. 4788, based on fatty acids. The fatty acid may be any of free fatty acids, c ₂ to C _24, preferably stearic acid. The organisms are preferably present in about the same amount, but their proportion can range from about 30 to about 70% of one of the organisms, the remainder being the other.

It is not known precisely why these two organisms provide the desired characteristics of the invention, particularly for poultry, in particular increased weight gain, improved feed conversion, increased breast meat yield and increased flock weight uniformity. Indeed, this is the case if both are present in a combination so that they can interact with each other, and provided that the amount is within the above range. It is through the interaction and interaction of these factors that the desired characteristics of the invention are achieved, which make it possible to significantly improve the composition, quality and processing of the meat in poultry. Similar results can be obtained in pigs as documented in the examples.

the fed microbial composition can vary considerably, but will range from about 0.5 to about 2.0 pounds per ton of about 1.2 pounds per ton of feed Pound per ton of feed. The number of organisms present was about 10,000

0,8 to

The amount directly added to the ration, it should be.Lo be in the feed, usually about and most often about 1

i the number of colony forming units per gram, in the probiotic, may also be in the range of up to about 2.10 < 3 > CFU / g and preferably about 2.10 < 3 > CFU / g.

If the above-described directly fed microbial composition is chosen for feeding in the feed ration, the combination of the above two strains behaves as a growth promoter. Growth promoters now used include antibiotics such as Sta6 p

fac and BMD. Advantages of subtherapeutic amounts of antibiotics as growth promoting ingredients can be achieved by the naturally occurring organisms of the invention when the directly fed microbial composition is produced in accordance with the invention and added to the feed by the method of the invention. Furthermore, some experiments have been carried out showing that the combined combination of the directly fed microbial composition and the growth promoter outweighs the advantages of each of the individual components and can therefore be used together if desired. In most cases, however, it is preferred to use the directly fed microbial composition alone, since one of the purposes of the invention is to eliminate the use of growth promoters at all.

The method of treatment of the organisms is not critical so long as the organisms can be kept alive until they are delivered to the animals and brought into a form such that they are well-fed to the feed and have a generally uniform size to control dosing.

A preferred means to meet these requirements is to process the organisms into microspheres with a fatty acid matrix. The microsphere refers to a fatty acid matrix into which various organisms are incorporated. It differs from microcapsules, in which individual organisms are always encapsulated. In the microsphere, the fatty acid matrix in the composite works similarly to the relationship between the biscuit dough matrix and chocolate chips, where chocolate chips represent groups of organisms. This procedure is described in Rutherford et al. In this procedure, the bacteria are combined with a heated fatty acid. The temperature of the fatty acid and the time of exposure of the bacteria to the fatty acid is controlled to keep the bacteria alive, but to be mixed with the fatty acid. The mixture is placed on a rotating disk to form microspheres containing bacteria where the fatty acid acts as a matrix. Several significant advantages are achieved using this method. First, the bacteria are kept alive during processing; secondly, the process in combination with the rotary disk technique allows to obtain a uniform size microspheres for better dosing. Third, the nature of the matrix, the β-fatty acid, allows the formation of unique microspheres. The combination of these factors results in a highly stable directly fed microbial composition with maximum efficiency.

In the process of the basic application, it is important to note that microspheres are formed where each ball is formed by a plurality of bacteria in a free fatty acid matrix rather than each individual bacterium being encapsulated in a coating or film-like layer of fatty acid. This brings the benefits of stability and more efficient dosing in bacterial processing.

The preferred matrix material is a free fatty acid up to C24. Mixtures of fatty acids may also be used, but it is preferable to use a single pure fatty acid. It is also preferred that the free fatty acid is an unsaturated fatty acid; most preferred is stearic acid.

Generally speaking, it is important that the fatty acid has a melting point of less than 75 ° C, preferably in the range of 40 to ° C. In order to function effectively as a matrix, it must of course be solid at room temperature. All free fatty acids within the chemical definition given above meet this requirement.

In order to increase the stability of the product, the bacteria are usually added to the product in lyophilized form. They can then be revived by adding moisture.

The microspheres produced as described below are typically from about 50 to more than 90% by weight of the fatty acid component and the remainder being a bacterial culture. The preferred range is about 60 to about 60. 75% fatty acid. If too little fatty acid is used, the matrix is not suitable for protection. In contrast, when using too much, the matrix is too thick and does not allow adequate release in the intestine.

The method of the invention uses a rotating disk to form microspheres. The rotary disc technology generally consists in mixing the bacteria-fatty acid suspension thoroughly and bringing the mixture to the center of the rotating stainless steel disc at the same speed. Due to the centrifugal force, the mixture is thrown outwards and forms microspheres. They are then collected in a refrigeration chamber, maintained at ambient or slightly lower conditions, sorted and prepared for packaging.

While encapsulation on rotary disks is known, it is not known to produce microspheres contained in a matrix without surrounding wrapper, nor to use microspheres or encapsulation in lyophilisations. lized bacteria. Encapsulation using a rotary disk is generally discussed by Johnson et al. of the Southwest Research Institute of San Antonio in the Journal of Gas Chromatography, October 1965, p. 345-347. A rotary disc device suitable for use in the present invention is described in detail in U.S. Patent 4,675,140, issued to Sparks on June 23, 1987, and entitled "Particle Coating Method for Liquid Droplets," referred to herein. Most preferred, however, is the process described in the basic application.

It should be emphasized that the rotational formation of microspheres provides a distinctly different product from both conventional spray drying and microencapsulation. In conventional spray tower drying, there is a tendency to agglomerate particles and form an irregular coating, which significantly affects the stability of the product, perhaps to a day to week level. A microencapsulation builds up a layer around the object and it has been shown that the bacteria are then too small, too difficult to keep alive or to obtain in a uniform size to be practically usable. In the manufacture of microspheres, in particular using the compositions described herein, the stability of the bacteria obtained, even when exposed to moisture and antibiotics, is achieved for three to six months, and the life of the bacteria in particles with uniform distribution remains.

If the free fatty acid microspheres of the invention are used within these ranges, the rotating disk, most often having a dimension of 4-6, can rotate at a rate of 2000 to 4000 min ^- 1, preferably about 2500 to 3200 min-1, and administration may be 50 to 200 g / min. The preferred conditions presently known are use of stearic acid, use of two of the above mentioned organisms, a four inch rotary disc, 3000 min ^- N, the rate of administration of bacterial suspension containing 35% bacteria, 65% stearic acid, 100 g / min. Under these conditions, a product with a particle size of 75 to 300 μιη, preferably below 250 μτη, is obtained.

BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1, 2 and 3 graphically show the stability of the strains using a stearic acid matrix.

Fig. 4 is a graph showing the distribution of breast meat in a feed experiment with a directly fed microbial composition according to the invention.

In FIG. 5 is a graph depicting body weight distribution in feeding experiments with the directly fed microbial composition of the invention.

In FIG. Figures 4 and 5 show a control experiment, an experiment using an antibiotic and an experiment using a directly fed microbial composition according to the invention.

DETAILED DESCRIPTION OF THE INVENTION

The invention is illustrated in more detail by the following non-limiting examples. Examples are described in connection with Figures 1, 2 and 3 in the accompanying drawings. Examples 1-4 and FIG. 1, 2 and 3 relate to an earlier invention of the same inventor. Example 5 and Tables 2 to 10 relate to the process of the invention for poultry feed directly. Example 6 relates specifically to turkeys and example 7 refers to pigs.

Example 1

Example 1 correlates with FIG. 1. Illuminates the stability of the product using two different Enterococcus faecium strains at 4 and 27 ° C. In FIG. 1 shows the stability of encapsulated strains of Enterococcus faecium after encapsulation using rotating disc devices using stearic acid and at a culture weight of 35%. Microspheres were formed under the above conditions, i. a suspension of bacteria in 35/65 stearic acid at 60 ° C using a 4-inch rotary disc with a rotation speed of 3000 rpm and a feed rate of 100 g / min. The beads were formed, placed in heat-sealed steam barrier bags and destructively sampled weekly for CFU determination. Obviously, the product of the invention retained excellent numbers of colony forming units (CFUs) for a storage period of up to 70 days.

Example 2

Example 2 is to be interpreted in conjunction with FIG. 2. The figure shows the stability of the individual strains in the form of microspheres when mixed with conventional feed in the presence of three

poultry antibiotics. feed a little Ingredients: finely crushed sweet corn 54% soya flour 26% fish meal 2% phosphate d i.alcium 1.5 % limestone 1% soybean oil 5.5 % moisture content 12% Bol i added three antibiotics v this amount: decochionate 6% (454 ppm) sal. spectinomycin (50 ppm) t and monensin sodium

(120 ppm).

Culture was added to the mixture in an amount providing approximately 1.10 µg CFU / g feed. The feed was packaged in heat sealed bags and incubated at room temperature. Weekly samples were taken for CFU determination. The graph of FIG. 2 shows excellent stability.

Example 3

Example 3 is to be interpreted in conjunction with FIG. 3. Illuminates the stability of Enterococcus faecium microspheres in the feed in the presence. various antibiotics. The feed consisted of 60% finely ground corn, 38% soy flour and 2% limestone with a moisture content of about 14%. The culture was added to a value of approximately 10 µCU / g feed and mixed. Ten pound aliquots were stored in sealed bags at 20 ° C and sampled weekly for 16 weeks. The amount of antibiotics included was:

bacitracin methylenedisalicylate 50 g / t carbadox 50 g / t chlortetracycline 200 g / t lasalocid 30 g / t 1ineomycin 100 g / t neomycin 140 g / t oxytetracycline 150 g / t Sulfamethazine 100 g / t tylosin 100 g / t virginiamycin 20 g / t ASP250 100 g / t furadox 10 g / t In Table 1 they are listed as a minimum times for loss 1 log colony forming units (CFU).

Table 1. Week time for loss of 1 log CFU at 20 ° C in kneaded feed with 14% humidity

antibiotic storage time (days) inspection 103 bacitracin 88 carbadox 54 chlortetracycline 60 lasalocid 57 1 incomyci. n 75 neomycin 53 oxytetracycline 59 sulfamethazine 62 tylosin 52 virginiamycin 112 ASP250 67 furadox 53

Example 4

In Example 4, the stability of the product after pelleting for use in chicken feed was determined. Microspheres were formed under the above conditions. In this experiment, the following conditions were used:

Crude protein, Not less than 18,0% Crude fat, Not less than 5,0% Crude fiber 6,0%

Pellets with and without antibiotic (CTC 50 g / t) were produced with the following ingredients and conditions: corn, SBM, whey, soybean oil, dicalcium phosphate, limestone, trace mineral premix. vitamin premix, selenium, copper sulfate. The culture was added at approximately 5.10 $ CFU / g feed. The ionization temperature a was 70 ° C and the pellets downstream were 78 ° C. The pellets were also stored in unsealed bags and weekly samples were taken for CFU determination.

In all cases, the stability of the product was not adversely affected by the pelletization conditions. In particular, the pelletized product had stability equal to that of the unpelletized product.

Example 5

The 560 day-old Peterson x Arbor Acres broiler chickens were randomly assigned to floored feeders (Table 2) with treated litter and fed for 45 days. All birds that died during the first 5 days were replaced by same-sex birds from the same van and treated. The composition of the basic nutrient initiation, growth and attenuation doses is shown in Table 3. These feed rations were formulated to contain 1,425 (initiation), 1,450 (growth) and 1,475 (attenuation) kcal ME / lb together with 90 g / ΐ monezine. The initiation dose was administered from day 1 to day 21, growth from day 21 to day 42, and sedation from day 42 to day 49. Individual experiments were evaluated: crushed negative control (control M); selected encapsulated directly fed microbial cultures containing Enterococcus faecium 301, no. DSM-Nr. 4789 and Enterococcus faecium 202, no. DSM-Nr. 4788, encapsulated with a fatty acid by means of a rotary disc according to Example 1 and present individually in an amount of 50% of the directly fed microbial composition delivered to the ground feed at a dose of 1.10 ⁵ CFU / g (probiotic M); pelletized negative control (control P); directly fed microbial composition in an amount of 1.10 µg CFU / g crushed feed, pelletized (directly fed microbial composition P) and positive control containing 10 g / t virginiamycin, pelletized (Stafac 10). The initial dose was crushed in the pelletizing experiments. Twelve twelve feed metal with 35 taps and 35 hens were used for each test dose.

Body weight, feed consumption and mortality after the first 5 days were recorded for each feed. Feed conversion, adjusted feed conversion and feed adjusted to body weight were calculated for each feed.

All data were scattered and differences were determined using Fisher's LSD.

Prior to the experiment, the directly fed microbial culture concentrate was adjusted with calcium carbonate. Theoretical calculations were for a directly fed microbial

Q

M 1.10 CFU / g product and, for the directly fed microbial composition P 2.10 CFU / g product. To determine the actual number, a 11 g sample was measured in duplicate for each product. Each sample was coated with the standard Pioneer method for lactic acid encapsulated bacteria.

A mixing test was performed for each production phase. The assay was designed to ensure uniform distribution of the directly screened microbial composition in the appropriate representation in the feed and to maintain its viability during pelletization. Samples were taken from each batch at the time of bagging; for crushed products 4 uniformly distributed samples and for pelletized products 10 uniformly distributed samples (i.e. bags 1, 3, 5, ..., 35, 37 and 39).

and 4.

In the first week of the experiment, samples of non-contaminated melting feeders were taken and samples of the feed experiment were taken.

During 1.

mined fodder feed; from weeks 2 and 6

An equal number of birds of both sexes were sacrificed to determine the weight of breast meat, body weight and small intestine weight and length. For each bird, chest meat yield and intestinal weight to length ratio were calculated.

All data were subjected to a scattering analysis and the difference was determined by contrast and estimated values of the desired effects.

Sixty birds from each experiment were delivered to the university for sensory appetite evaluation.

Directly fed microbial irrespective of processing improved (P < 0.05) feed conversion over the respective control, while weight gain increased (P < 0.05) over control only for crushed feed (Table 4). Directly fed microbial composition P improved (P> 0.05) feed conversion over Stafac ® 10, which had a similar value (P> 0.05) to control P.

The product had the desired composition in terms of representation and strain (Table 5).

The directly fed microbial composition was evenly distributed throughout the feed. The directly fed microbial composition M had the desired representation, while the directly fed microbial composition P was 1 to 11 1/2 logs higher than required for (Table 6). The high numbers of Formulation P resulted from the assurance of sufficient yield of an initiation and growth feed dose of directly fed microbial oversizing of the product for pelletization organisms.

of the floor feeders in the number of 7). Directly fed microbial growth and attenuation mixtures in 4.2 log.

In the case of directly fed microbial P, the samples corresponded to the shuffling test (composition table and week 6 i).

However, M fed (P <0.05) versus control M as the weight of him meat (Table 8), composition P showed

Improved even earlier

Thus, directly in the crushed microbial composition M, the chest yield increased while the directly fed microbial improvement (P > 0.05) compared to the control of P. feed agrees with the results of the experiment. Directly fed microbial composition of similar magnitude to improve breast meat yield,

P did not show, as in the case of directly fed microbial composition M. to be a consequence of leaves

This failure can make pelletization that better energy use less room for improvement.

Pelletize with

P ^r iemerná weight of feed has increased by 96 dock increased uniformity improvement in the mash feed.

gP r i amo fed bird weights bird versus shredded microbial (Fig. 5)

Pelletization increased the abrasion and the weight of the breast meat by means of the meat compared to the meat and uniformity (fig.

g. Directly fed microbial control average chest weight4) with the greatest improvement in crushed10 showed the greatest improvement in average uniformity in pelleted feed.

Pelletization yielded 0.53 percent units compared to ground feed. The directly fed microbial composition M showed an increase of 0.84 percentage units over the control M, which was similar in size to the pelletization result.

Treatment with the directly fed microbial resulted in a shorter (P> 0.05) small intestine length than in both R controls and Stafac, expressed in actual length, body weight to breast meat ratio (Table)

9). The directly fed microbial composition M had a smaller (P > 0.05) small intestine weight than the control M, expressed as either actual weight or percentage of body weight or weight of breast meat. The reduction in bowel weight and length in the case of a directly fed microbial formulation indicates less nutrient energy and more energy available for growth, as evidenced by improved feed conversion and breast meat yields (Tables 7 to 8).

Birds treated with directly fed microbial P did not have 10 foreign odors compared to Stafac (table).

10). In a second experiment, it was found that directly fed microbial composition P improved the taste of the thighs over control P. However, this improvement was not observed in the first attempt.

Table 2. Feed Metal Layout Treatment Feeder Number Control P Product P Stafac 10 Control M Product M

2.6.15.17.22.26.104.109.113.117.122.126

4,8,12,16,21,28,105,106,112,118,125,130

5,7,11,18,23,27,101,107,111,116,123,129

3.9.13.20.24.30.102.108.114.119.121.127

1,10,14,19,25,29,103,110,115,120,124,128

4.2 'x 15.5' feed size, one pipe feed, one suspended watering feeder, pine shavings as litter, energy and evaporative cooling system, well insulated forced air heating, curtain wall design.

Table 3. Composition of basic feed

Ingredient A production phase initiatory growth attenuation ground corn 65.37 % 67.89 74.29 soya flour 25.58 23,53 17.83 meat - and - bone meal 3.00 3.00 3.00 grease 3.36 3.32 2.59 defluorinated phosphate 0.95 0.79 0.73 calcium carbonate 0.61 0.62 0.63 salt 0.35 0.31 0.32 trace minerals 0.05 0.05 0.05 met i on 0.39 0.28 0.33 lyží n 0.19 0.06 0.18 Vitamin Oremix 0.05 0.05 0.05

Table 4. Production data for floor feeders

Pelleted Crushed

inspection P * Stafac lO ^ inspection M * weight, lb 4.79 ^a 4.81 ^a 4.79 ^a 4.54 ^b 4.68 ^and _L feed conversion 1,871 ^b 1,827 ^a 1,855 ^ab 1,917 ^c l, 856 ^{and b} adjusted feed conversion ^{1 2 *} 1,832 ^b 1,789 ^a 1,807 ^ab 1,887 ^c 1,812 ^ab feed conversion • modified to 2 weight 1,801 ^b 1,755 ^a 1,775 ^ab 1,897 ^c 1,798 ^b « mortality rate,% 4.40 4.64 5.95 3.33 5.60

adjusted feed conversion = total feed / (live + dead weight) feed converted to weight = adjusted feed conversion - ((weight-4.60) / 6) ^abc P <0.05 * according to the invention

Table 5. Product quality control and assurance

Treat ren i. and QC number Number of QA ⁷ Strain ratio means P ----- ^CF U / g of product 5.75.10 ° 1.01.10 ⁸ SF202: SF301 50:50 means M 9,54.10 ^z 9,60.10 ' 57: 43

quality control

Table 6. Feed mix test and yield

Production phase and treatment crushed pelleted yield ^ --- CFU / g product --- % crushed initiatory control P NA ² 1,06.10 ³ - means Stafac ^ 10 P 2,02.10 ⁶ NA 1,67.10 ° 6,46.10 ³ 98.69 control M 2,51.10 ³ means M 1,34.10 ⁵ growth control P r grid Stafac ^ 10 P NA 3,89.10 ⁶ 5,25.10 ⁴ 4.86.10 ² 1.09.10 ⁶ 6.42.10 ³ 91.62 control M 1,00.10 ² resource M 1,48.10 ⁵ Út l.mová control P 8,50.10 ² 1,11.10 ³ - environment Stafac 10 P 7,04.10 ⁴ 4,91.10 ⁵ 117.40 8,80.10 ³ 1,79.10 ⁴ - control M 8,92.10 ² p ršt r edok M 1,33.10 ⁵ The middle control P 8,50.10 ² 8,28.10 ² - Stafac 10 P 8,21.10 ⁵ 2,15.10 ⁴ 9,64.10 ⁶ 9.05.10 ³ 118.09 control M 8,72.10 ² p ršt r edok M 1,38.10 ⁵

yield calculated from data transformed into log? g

NA = not available

Table 7. Quality assurance in floor feeders ____________ Week ____________________ Med

Treating ________1_________2__________4__________6_______________

-------------------- C F U / g feed and ------------------

kont rola 3,78.10 ~ 3,83.10 ~ 8,60.10 ^ 2,21.10 ^z 4,08.10 means P 9,23.10 ⁵ 9,37.10ζ 8,77.10 ³ 8,48.10 ⁵ 8,96.10 Stafac 10 8,73.10 ² 1,29.10 ² 6,46.10 ² 8,63.10 ² 8,89.10 control M 3,46.10 ² 1,26.10 ² 2,79.10 ³ 2,00.10? 5,08.10 means M 1,43.10 ⁵ 1,25.10 ⁵ 1,75.10 ³ 1,00.10 ³ 2,32.10

Table 8. Evaluation of breast meat yield

crushed

pelleted

inspection P * Stafac lO ^ inspection M ' body weight, g 2,240.7 2,230.1 2,195.9 2,143.8 2,149.9 mass of breast meat, g 234.4 ^a 239,6 ^a 232.0 ^a 213.3 ^pts 229.6 ^a breast meat yield,% body weight. 10.51 ^a 10.68 ^a 10.58 ^a 9.93 ^b 10.67 ^a

ab

P < 0.05 * according to the invention

Table 9. Intestinal weight and length ________ Pelleted _____ Crushed control P * Stafac ^R 10 control M * body weight

g 2,240.7 2,230.1 2,195.9 2,143.8 2,149.9 mass clod - meat, g 234.4 ^a 239,6 ^a 232.0 ^a 213.3 ^pts 229.6 ^a weight TS, g 92.6 93.3 93.4 91.4 87.4 length TS , i. n 76.3 75.3 76.6 76.1 75.3 TS, g / in 1.21 1, 23 1.22 1.20 1, 16 weight TS, g / 100 g of body weight i 4.17 4.18 4.27 4.29 4.08 length TS , in / 100 g of body weight t i 3.47 3.40 3.53 3.61 3.53 mass TS, g / 100 g weight i h ore meat 40,19 39.70 40,97 43,96 38,69 length TS , in / 100 g weight t i thoracic meat 33,41 ^a 32,27 ^a 33.72 ^a 36.89 ^pts 33.41

^ab P <0.05 according to the invention TS small intestine

Table 10. Taste panel evaluation

Count right IDs tissue Group comparison Experiment 1 Experiment 2 sum thigh D Stafac 10 control P 6 3 9 Stafac ^R 10 XINOC P 3 4 7 means P vs. control P 2 8 * 10 chest Stafac ^ 10 control P 2 6 8 Stafac ^R 10 XINOC P 1 3 4 means P vs. control P 5 4 9

evaluators were able to detect a single sample in a statistically significant (P <0.05) number of cases ^{1 the} number of correct individual sample identifications required for significance at 5% was 7 for n = 10 and 11 for n = 20

An experiment with broilers was performed to determine the efficacy of the directly fed microbial composition in the pulverized and pelleted feed. The directly fed microbial composition, irrespective of processing, improved (P <0.05) feed conversion over the corresponding control while increasing (P <0.05) weight gain over the control only for crushed feed. Directly fed microbial

D

P improved (P> 0.05) feed conversion over Stafac 10, which had a similar result (P> 0.05) to control P. Directly fed microbial M increased (P <0.05) versus control M both weight and weight. breast meat yield, while the directly fed microbial composition P showed an improvement (P> 0.05) over control P. Birds treated with the directly fed microbial composition P had no foreign flavor compared to Stafac 10 treatment.

Example 6

144 mixed fed pigs (mean starting weight 41.5 lb) were randomized by weight and sex into pens (Table 11) with slate floor and fed 119 days. The basic growth and final dose composition is shown in Table 12. Growth doses were administered to an average weight of 120 lb, followed by slaughter followed by final doses. All doses up to 75 lb body weight contained Mecadox (50 g / t) and then up to 120 lb body weight 100 g / ΐ chlortetracycline. Treatment of animals was negative (control) and selected directly fed microbial cultures in the form of microspheres, administered in an amount of 1. 10 ⁴ CFU / g feed. All feed rations were administered in a crushed form. Six double pens of 12 feed pigs were used for each experimental feed.

Upon arrival at the research facility, pigs were given D

Ivomec against internal and external parasites. After four D weeks, Safeguard was administered against the nematodes.

Body weights, feed consumption and mortality were recorded for each pen. Feed conversion was calculated for each pen.

Prior to the experiment, the culture concentrate in the form of microspheres 7 was adjusted with calcium carbonate. The theoretical number was 2.10 CFU / g of product. A duplicate 11 g sample was investigated to determine the actual number. The sample was coated with a standard coating method for bacteria in lactic acid microspheres.

In addition, a 1 g sample was tested in duplicate to verify the number and composition of strains in the product.

For each treatment, samples were taken once a week and tested for bacteria in microspheres with lactic acid.

The desired number of organisms was confirmed for the product (Table 14).

Pen sample recoveries varied from 1:10 to Ι ^, ^ ό.ΙΟ cpy / g _rm IVA (Table 15). Both extreme samples were assigned to error errors. collection or coating. The rest of the samples were. evenly distributed around a target value of 1.1 () 4 (jpu / g feed).

The product as microspheres improved (P> 0.05) weight gain and feed conversion over the 28-day control (Table 13). In the first week of the experiment, pigs were affected by the onset of TGE. This, together with the time it takes to adapt the gastrointestinal tract of the pigs to the product, may result in a 28-day delay in the observed response. The results show that the microspheres of the directly fed microbial composition of the invention, as well as for chickens and guinea pigs, function efficiently for pigs.

Table 11. Cot Layout Experiment 670-9102

treatment pens numbers inspection directly fed pros th redok 3, 4, 6, 9, 11.12 1, 2, 5, 7, 8, 10 Size of the pen 4.62 ’x 16.0’, one four - hole

Smidley, one suction feeder, sprinkler-controlled temperature, partly slate floor and landscaped building with open front wall

Table 12. Composition of basic feed

ingredients Production phase growth Reduced ground corn % 76.30 82,20 soya flour 21.25 15.50 dicalcium phosphate 1.05 0.90 calcium carbonate 0.85 0.85 salt 0.30 0.30 vitamin and mineral premix 0.25 0.25

Table 13. Production data for pens, Experiment 760-9102

inspection Directly fed product Day 14 growth weight b 9.6 9.2 conversion feed 2,439 2,483 mortality % 1.41 1.39 Day 28 increase weight lb 26,3 27.9 conversion to rm i va 2,405 2,212 mortality % 2.82 1.39 Day 42 while growing weight i , Lb 45.8 46.8 conversion i a k rm.i va 2,497 2,428 mortality % 2.82 2.78 Day 56 growth weight i, lb 69.1 71.5 konve rz i a to rm i va 2,507 2,457 mortality % 4.23 2.78 Day 70 while growing weight lb 89.4 91.2 konve rz i a to rm i va 2,735 2,674 mortality % 5.63 2.78 Day 84 growth weight lb 111.0 112.0 watering can k r m.i va 2,904 2,882 mortality % 5.63 2.78 Day 98 while growing weight lb 129.9 134.7 conversion feed 3,071 2,988 mortality, % 5.63 2.78 112. day growth weight lb 152.2 154.7 konve rz i a to rm i va 3,164 3,134 mortality % 5.63 2.78 119th day while growing weight lb 162.4 165.6 conversion k rmiva 3,217 3,177 mortality % 5.63 4.17

Table 14. Product quality control and assurance, experiment

670-9102

Product Number of QCs Number of QAs ratio strain directly fed microbial --- CFU / g 4.3 Product— .10 ⁷ SF202: : SF301

Table 15. Quality assurance in pens, Experiment 670-9102

The date inspection Directly fed product 03/05/91 1,7.10 ³ 7.4.10f 05/08/91 1,8.10 ⁴ 1.6.10 ⁵ 22/5/91 0 1,0.10 ¹ 30/5/91 about 2,6.10 ³ 05/06/91 2,0.10 ² 2,2.10 ⁴ 12/6/91 1,0.10 ¹ 2,0.10 ⁴ 19/6/91 0 9,6.10 ³ 26/6/91 6,7.10 ² 5,6.10 ³ 03/07/91 4,9.10 ² 3,2.10 ³ 10/7/91 5,2.10 ¹ 3,0.10 ⁴ 17/7/91 1,0.10 ² 4,5.10 ³ 24/7/91 0 1,0.10 ⁴ 31/7/91 0 1,2.10 ⁴ 07/08/91 0 6,4.10 ⁴ 14/8/91 0 7,7.10 ³ 21/8/91 0 1,2.10 ⁴ 28/8/91 0 9,1.10 ³ 04/09/91 0 2,9.10 ⁴ 18/9/91 0 8,9.10 ³ 25/9/91 0 5,5.10 ³ The middle 9,5.10 ° 8,4.10 ³

/ to 7 7 7 b - 7 ¥

Claims (16)

PATENT CLAIMS A method of promoting animal growth, comprising adding to the feed an effective amount of a directly fed microbial composition consisting of substantially dried viable stable microspheres of Enterococcus faecium 301, no. ATCC 55059, fatty acid-based and dried viable stable microspheres of Enterococcus faecium 202, no. ATCC 53519, fatty acid 1.in. Method according to claim 1, characterized in that the fatty acid microspheres are formed by means of a rotary disk. The method of claim 2, wherein the directly fed microbial composition is about 30 to about 70% of one and the remainder is the other of said fatty acid microspheres. 4. The method of claim 3 wherein the fatty acid is a free fatty acid, C ^ 24 ^- The process according to claim 4, wherein the fatty acid is stearic acid. The method of claim 1, wherein the amount of directly fed microbial composition added to the feed is about 0.5 pounds to about. 2.0 lb / iva of feed. The method of claim 6, wherein the amount of directly fed microbial composition is about 0.8 to about. 1.2 Ib / t of feed. The method of claim 6, wherein the number of organisms directly fed the microbial composition is about. 1.10? To about 2.10? CFU / g. The method of claim 8, wherein the number of organisms directly fed with the microbial composition is about ^{10 5} CFU / g. 10th Chicken method. pod ľa Claim 1 characterized by that the animal is 11. process by Claim 1 characterize yourself by that the animal is pig. 12th Directly fed microbial growth promoting agent comprising essentially dried viable stable microspheres of Enterococcus faecium 301 ATCC no. 55059 fatty acid-based and dried viable stable microspheres of Enterococcus faecium 202 ATCC no. 53519 fatty acid. A directly fed microbial composition according to claim 12, characterized in that the fatty acid is free fatty acid C 1 to C 24. The directly fed microbial composition of claim 13, wherein the free fatty acid is stearic acid. 15. The directly fed microbial composition of claim 12, wherein the animal is a chicken. A directly fed microbial composition according to claim 12, wherein the animal is a pig. 17. The method of claim 1, wherein Enterococcus faecium 301 and Enterococcus faecium 202 are present in about equal amounts. A directly fed microbial composition according to claim 12 comprising about 20 to about 30% one and the remainder being the other of Enterococcus faecium 301 and Enterococcus faecium 202. A directly fed microbial composition according to claim 16, characterized in that the microspheres of Enterococcus faecium 301 and Enterococcus faecium 202 are present in about equal amounts.