HUT67965A - Fatty acid microspheres containing enterococcus for use to enhance growth and improove carcass qualty - 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

Α Growth promoters, widely used in the form of antibiotics in poultry, namely chickens and turkeys. Growth enhancers such as Stafac and BMD (bacitracin methylene disalicylate) are well known antibiotics and are used at the sub-therapeutic level, for example as a nutritional supplement at 10 grams per tonne and 25 grams per tonne to promote the desired growth characteristics of chickens. . However, the use of antibiotics for this purpose has recently been criticized. On the one hand, the criticism is that the bird eventually develops tolerance to antibiotics and as a result the antibiotic is no longer capable of increasing growth. Other objections are of a health nature that concern the use of unnatural antibiotic supplements and their effects on development. However, due to the benefits of using antibiotics, they are still widely used with the aim of improving nutritional utilization, improving body composition and enhancing growth.

It is well known that certain microorganisms, such as bacteria, have the potential benefit of being administered to animals. Bacteria are useful, for example, because of the natural microflora of the intestine. Some companies provide nutritional supplements and sell microbial products with beneficial properties that can be used directly as nutrition. However, the problem with microorganisms introduced directly into food is that they have problems with the stability of the stored product. Typically, the microbial product that can be used directly as food is added to the diet in relatively small amounts, about 0.1% -2-.

in. However, a nutritional or nutritional supplement product containing an unused microbial product that can be used as direct food is often stored by farmers for a long time. This storage often means conditions where all moisture is present. In many cases, the moisture is just enough for the bacteria to activate or start to grow, but not enough to ensure growth. As a result, they die. Thus, the activity of the bacterial product used as a direct food disappears. In other cases, the addition of antibiotics to the microbial product or nutritional supplement used as a direct food will adversely affect the bacteria, especially when a small amount of moisture is present and the bacteria will be killed. Thus, long-term storage of microbial products that can be used as direct food raises significant problems.

In other environments where microbial products for direct nutrition are also used, such as chicken feeds, it is common practice to granulate the material with the microbial product added prior to granulation. The steam used in the granulation partially activates the bacteria, which kills them due to the lack of moisture in their livelihood. The heat applied during granulation can also kill them. In addition, there is an acidic environment in the stomach that potentially inactivates the bacteria before they reach the intestines. So there is a constant need for a microbial product for direct nutrition that will release the microorganisms into the intestines at just the right time without having to •

they would be released because of the wet conditions or the unfavorable pH conditions that prevail in the digestive tract before the small intestine.

It is particularly desirable to achieve, if possible, certain characteristics of the birds. These include faster weight gain, better nutrition, better composition of body material and, finally, greater body weight of animals. Of course, the higher rate of weight gain and better nutrition are important because of the economic aspects that accompany these desirable results. The composition of the body material is also important, as the most important place for tissue deposition is the breast, as this yields the largest amount of clean meat. So not only is weight gain important, but also where the body is gaining weight. The weight of each animal is also important, as having more birds of normal size will require less manual labor and more efficient processing by machine. On the other hand, if the size of the birds varies significantly from small to very large, although the size of the population may be the same, smaller and larger birds require more manual labor and, due to the lack of uniform size, are not easily machine-processed them. Thus, one desirable feature is that the size of the birds should be uniform and within a large percentage of the normal size range, so that the chickens can be processed using standardized equipment.

Similarly, it is a microbial product which can be used as a direct food and not only for poultry, ie chickens and turkeys, but also for pigs.

-4, would be very beneficial. It is a primary object of the present invention to provide a microbial product that is directly fed to chickens which does not contain antibiotics, but microspheres that occur naturally.

The present invention relates to a usable microbial

microorganisms with furthermore direct as food product Production and which is two

microorganism, namely Enterococcus faecium 301 (DSM

No. 4789) and Enterococcus faecium 202 (DSM No.

4788).

DSM stands for Deutsche Sammlung von

Microorganism (Braunschweig,

FRG).

These microorganisms were deposited with the ATCC.

The present invention further relates to a process for the production of a microbial product for use as a direct feed, which provides increased weight gain in poultry, provides better nutrition, provides greater weight to the breasts, and provides the animals with a uniform body weight within the normal size range.

The present invention relates to the production of microbial products for direct nutrition which are suitable for addition to poultry feeds and which contain microorganisms contained in microspheres produced by a special rotary disk technique using free fatty acid to form spheres.

The present invention further relates to the production of a microbial product for direct nutrition which has a stability of between 3 and 6 months without significantly reducing the number of microorganisms during this time.

-5 • ·· · ·· «·« ·

The present invention further relates to a process for rotationally producing microspheres of dried bacteria, characterized in that the spheres are of uniform size.

The present invention also relates to the production of rotating disk microspheres of dried bacteria which are free flowing and can be easily processed with poultry feeds.

The invention further relates to the production of microspheres of certain bacteria made from fatty acids, which are suitable for direct feeding of both poultry and pigs.

The description of the accompanying drawings is briefly described below.

Figures 1, 2 and 3 graphically show the stability of microspheres of strains using stearic acid as the free fatty acid.

Figure 4 is a graph showing the distribution of breast meat obtained in a feeding experiment with a microbial composition for use as a direct food according to the present invention.

Figure 5 is a graph showing the body weight distribution obtained in a feeding experiment with a microbial composition for use as a direct food of the present invention.

Figures 4 and 5 also show a control with the use of an antibiotic and a microbial product of the present invention for use as a direct food.

The present invention relates to a method and composition for promoting the growth of poultry and pigs, comprising administering to the normal diet a small but growth-promoting amount of a microbial product which can be used as direct food, Enterococcus faecium 301 (DSM No. 4789) and Enterococcus faecium 202 ( DSM No. 4788) contains a dried culture of fatty acid encapsulated in microspheres, wherein the fatty acid microspheres are prepared by rotary disk drying.

It has been a surprising discovery that growth enhancement of poultry and pigs can be achieved by adding dried fatty acid microspheres of Enterococcus faecium 301 (DSM No. 4789) and Enterococcus faecium 202 (DSM No. 4788) to the normal feed. The microsphere preparation is preferably used for carrying the C ₁₂ ^-C 24 free fatty acid, but it is preferred if the fatty acid is stearic acid. The microorganisms are preferably present in about the same amount in the composition, but may be present in a proportion of 30-70% for one microorganism while the remainder is provided by the other microorganism.

It is not known exactly why these two microorganisms provide the desired properties of the present invention, particularly in poultry, i.e., increased weight gain, improved nutrition, increased lean meat weight, and animal body weight uniformity. The fact is that they are provided on the condition that both are used in the composition and therefore may interact with each other in some way and provided that they are used in said amounts. It is the interaction and coefficient of properties that provides the properties of the present invention.

-Ί desirable qualities and that provide significantly better quality of body, meat and processability. Similar results can be obtained with pigs, as can be seen from the examples.

The amount of microbial product that can be used as a direct feed can vary widely, but is generally used in a ratio of 0.25 to 1 kg / tonne feed, typically about 0.4 to 0.6 kg / tonne feed, and typically about 0.5 kg / tonne. tons of feed in proportion. The number of organisms present in the number that is used as a direct fed microbial product as colony-forming units can vary between 1 x 10 θ CFU / g to 2 x 10 ⁹ CFU / g, preferably from about 2 x 10 ⁸ CFU / g.

When the microbial product for direct feeding described above is added to the animal feed, the two strains of microorganisms act as growth promoters. Current growth promoters include antibiotics such as Stafac and BMD. The benefits of sub-therapeutic antibiotics as growth enhancers can be achieved by the naturally occurring organisms of the present invention, provided that the microbial product used as a direct food is prepared by the method of the present invention and administered to the animal according to the method described. supply power. Indeed, there have been a few studies that suggest that the combination of a microbial product as a direct food and a growth promoter can enhance the benefits of individual applications, so that they can be used together when needed. In most cases, however, it is preferable to use only a microbial product suitable for direct feeding, · · · · · · · · · · · · · · · · · · · ·

It is an object of the present invention to avoid the use of growth enhancing agents.

The method used to process the microorganism is not critical, as long as the microorganism is alive to be delivered to the animal and formulated so that it can be properly combined with the animal diet and generally of a uniform size so that dosage can be controlled.

A preferred means of meeting these requirements is to place the microorganisms in a fatty acid carrier. The microsphere refers to a fatty acid carrier that contains many different microorganisms. This is different from the microcapsule in which the individual microorganisms are individually encapsulated. In a microsphere, the fatty acid carrier acts relative to the whole composition, such as in the chocolate donut, the relationship between the donut and the chocolate, and chocolate refers to microorganisms. The process is described in the first patent application by one of the inventors, Rutherford et al. In this process, the bacteria are contacted with a heated fatty acid. The temperature of the fatty acid and the time the bacterium is in contact with the heated fatty acid are controlled to keep the bacterium alive and still be able to mix with the fatty acid. The mixture is applied to a rotating disk which results in the formation of bacteria encapsulated in the microsphere, the fatty acid being used as a carrier. Applying this method has several important benefits. First, the bacteria survive the procedure; second, the combination of the rotary disk and the microspheres of the same size allows for better dosing.

• · ·

-9 Thirdly, the nature of the carrier, the fatty acid, allows us to create unique microspheres. The combination of factors results in a very stable microbial product that can be used as a direct food, with maximum efficiency.

The previous. In the process of the original patent application, it is essential that microspheres are formed, each of which contains a large number of bacteria in a free fatty acid carrier, and not microencapsulation, where each bacterium is coated with a fatty acid coating or film-like layer. This has the advantages of stability and more effective dosing during bacterial treatment.

The preferred carrier for C ^_42-c 24 fatty ^Patent d. Although a mixture of fatty acids may be used, it is preferable to use a single pure free fatty acid. It is also preferred that the free fatty acid is a saturated fatty acid, with stearic acid being most preferred.

In general, it is preferred that the fatty acid has a melting point lower than 75 ° C, preferably between 40 ° C and 75 ° C. Of course, it must be solid at room temperature to be an effective carrier. All fatty acids that meet the chemical specifications given above meet these requirements.

In order to enhance the stability of the product, the bacteria used in the product are typically freeze-dried bacteria. So they can be brought to life by adding moisture.

In microspheres prepared according to the procedure described below, the particles are generally 50-90%

-10 contain the fatty acid component, the remainder is a microorganism, usually a bacterial culture. Preferably, the proportion of fatty acids is 60-75%. If too little fatty acid is used, the cover is not sufficient for protection. On the other hand, if used too much, the coating will become too thick, resulting in the inadequate release of microorganisms in the stomach.

The microsphere process used in the present invention is a rotating disk process. In rotary disk technology, microorganisms, often bacteria, and fatty acid components are thoroughly mixed and then transferred to a center of a rotating stainless steel disk at a uniform rate. The particles are scattered outward due to the centrifugal force. It is then collected in a refrigerator which is kept at room temperature or slightly lower, classified according to size and prepared for packaging.

Although the rotary disk encapsulation process is known, it is not yet known to be used to make microspheres contained in a carrier without an envelope surrounding it, nor is it known that the microsphere process or encapsulation was used with freeze-dried bacteria. The method of encapsulating substances by rotary disk method is described by Johnson et al., October et al., October 1965, Johnson et al., Southwest Research Institute of San Antonio, Journal of Gas Chromatography 345-347. In addition, a rotary disk apparatus useful in the process of the present invention is described in detail in United States Letters Patent 4,675,140 (June 23, 1987);

-11 • · «·· * · · ············································ · Methods Method Coating Particles or Liquid Droplets. The above publication is hereby incorporated by reference. However, the most preferred method is the one described in the previous patent application.

It is important to note that the rotary disk based microsphere preparation process results in a completely different product than conventional spray tower drying. In conventional spray tower drying, the particles have a tendency to aggregate as they are not uniformly coated and consequently the product stability varies from day to week.

By doing this, we get a shell around an object and the bacteria have been proven to be too small, too hard to survive, and to have a practically uniform size. Bacteria produced in the microsphere live environment remain stable for three to six months, even when exposed to all moisture or antibiotics, and retain bacterial viability in uniformly distributed particles.

If the free fatty acid carrier of the present invention is used within the limits described below, a process using a typically 10-15 cm rotating disk may be performed at 2000-4000 rpm, preferably 2500-3200 rpm, at a feed rate of 50 rpm. -200 grams per minute. The currently known preferred parameters are the use of stearic acid and the two previously mentioned microorganisms, a ten centimeter rotating disk, 3000 rpm, and a feed rate of 100 grams per minute.

-12 • · «· · * * * perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc perc. From a mixture containing 35% bacteria and 65% stearic acid. Under these conditions, a product consisting of particles having a diameter of 75-300 microns is obtained, preferably less than 250 microns in size.

★★★★★★★★★★★★★★★★★★★★

The following examples are provided to further illustrate the process of the present invention without limiting its scope. Some examples are given in connection with Figures 1, 2 and 3. 1-4. Examples 1 to 4 and Figures 1, 2 and 3 relate to the previous patent application. Example 5 and Examples 2-10. Tables 1 to 5 relate to the method of feeding the chickens of the present invention with the direct microbial product. Example 6 refers to turkeys and Example 7 relates to pigs.

Example 1

The

Example 1 corresponds to Figure 1. This one

The stability of the product with two different strains of Enterococcus faecium is shown

At 4 ° C and

27 ° C. As shown in FIG

Shows the stability of microspheres of Enterococcus faecium strains prepared by rotary disk procedure with stearic acid from 35% by weight of culture. The conditions are as described above, i.e. a bacterial / stearic acid ratio of 35/65 at 60 ° C, use of a 10 cm rotating disk at 3000 rpm and a feeding rate of 100 grams per minute. The culture is coated with the support medium to form microspheres and placed in heat-sealed, moisture-resistant bags and subjected to destructive sampling ········

-13 ·· · «··» ·· ·· · CFU value is determined weekly. It can be seen that the product of the present invention holds the CFU count very well up to a storage time of 70 days.

Example 2

Example 2 is interpreted in conjunction with Figure 2. The figure shows the stability of each encapsulated strain when a typical feed. in the presence of three wing antibiotics. The composition is as follows:

54% fine-grained corn 26% wool ali 2% fish meal 1.5% dicalcium phosphate 1% chalk 5.5% weave oil 12% humidity

Three antibiotics are added in the following weight ratios:

decoquinoate 6% (454 ppm), Salinomycin (50 ppm) and monensin sodium (120 ppm).

The culture is added to the mixture at a concentration such that it has a concentration of about ΙχΙΟθ CFU / gram of feed. The medium was packaged in heat-sealed pouches and incubated at room temperature. Weekly samples are taken for CFU determination.

The graph in Fig. 1B demonstrates excellent stability.

Example 3

Example 3 is interpreted in conjunction with Figure 3. This shows the stability of the encapsulated Enterococcus faceium mixture a

-14 ·· 4 · · · · · · · · · · · · · · · · · · · · · · · · · · · · ··· ·

in food, different in the presence of antibiotics. The composition is

following:

60% corn flour 3 8% wool ali 2% chalk approx. 14% humidity

The culture was added to the mixture at about 10 CFU / g feed and mixed. 4.5 kg aliquots were stored in sealed bags at 20 ° C and sampled weekly for 16 weeks. The antibiotics were added in the following amounts:

Bacitracin methylene disalicylate 50 g / tonne carbadox 50 g / tonne chlortetracycline 200 g / tonne lasalocid 30 g / tonne Lincomicyn 100 g / tonne neomycin 140 g / tonne oxytetracycline 150 g / tonne sulfamethazine 100 g / tonne tylosin 100 g / tonne virginiamycin 20 g / tonne ASP250 100 g / tonne Furadox 10 g / tonne

Table 1 shows the minimum times to decrease the number of colony forming units (CFUs) by an order of magnitude.

··· «···· • ·

Table 1

Time taken for the order of magnitude decrease of CFU number in days at 20 ° C, 14% moisture

Antibiotic storage control 103 bacitracin 88 carbadox 54 chlortetracycline 60 lasalocid 57 Lincomicyn 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 is determined after granulation for use as chicken feed. The microencapsulation conditions are the same as described above. The conditions used in this test are as follows:

Crude protein, not less than 18,0%

Crude fat, not less than 5,0%

-16Nyers fiber, not more than 6,0%

Antibiotic (CTC 50 g / ton) granules with or without antibiotics are prepared with the following additives under the conditions indicated below.

Corn, SBM, whey, soybean oil, dicalcium phosphate, chalk, trace element premix, vitamin premix, selenium, copper sulfate. The culture is administered about 5xl0 ⁵ CFU / gm feed amount (100-150 gm / ton).

The conditioning temperature is 70 ° C and the non-paint granules are at 78 ° C.

The granules are stored in sealed bags and sampled weekly for CFU determination.

In no case is the stability of the granulated product adversely affected by the granulation conditions. More specifically, the stability of the granulated product was the same as that of the non-granulated product.

Example 5

Four thousand five hundred and sixty-one-day-old Peterson x Arbor Acres broiler chickens were randomly distributed into ground cages (Table 2) containing regenerated litter and fed for 45 days. Each bird killed during the first five days is replaced by a bird of the same sex from the same consignment and received the same treatment. Table 3 shows the composition of the base start, ascend, and withdrawals. Starting, growing and withdrawal feeds were formulated to contain 2850, 2900, and 2950 ME / kg at 90 g / tonne monensin. Starting rations were given at the age of 1-21 days, the female loser 21 • · ·

-17- ............

days, withdrawals at 42-49 days. The treatments were: negative control, meal mix (control, M); selected, encapsulated cultures of direct-feed microorganisms containing strains of Enterococcus faecium 301 (DSM No. 4789) and Enterococcus faecium 202 (DSM No. 4788), each encapsulated by the microbial direct nutrition method as described in Example 1; The composition was present in 50%, and they were used 1 x 10 ⁵ CFU / g feed amount (microbial cultures applied direct fed, M); negative control, granulated (control, P); microbial preparation for direct feeding, 1 x 10® CFU / g meal mix, granulated (microbial preparation for direct feeding, P); and a positive control, 10g / tonne virginiamycin, granulated (Stafac 10). The initial feed ration was broken due to granular treatments. Twelve parallel cages with 35 roosters and 35 hens were used for each experimental feed.

After the first five days, body weights, nutritional utilization, and mortality rates were recorded. Nutrient utilization, corrected nutritional utilization and weight corrected nutritional utilization were calculated for each cage.

All data are subjected to analysis of variance and differences are determined by Fisher LSD.

Prior to the assay, calcium carbonate was added to the microbial culture concentrate for direct feeding. The number of theoretical colony forming units of the microbial product to be used as direct food is 1 x 10® at M, 2 x at P • · · · ·

-18 ^- ............

Was 10 CFU / g. An aliquot of 11 g of each product is tested in duplicate to determine the actual CFU of the product. Each sample is plated using the standard plating technique described for encapsulated lactic acid bacteria.

A mixing test was performed at each production stage.

The test was designed to ensure that the microbial product used as a direct feed was evenly distributed in the feed and survived the granulation. Each batch is sampled prior to packaging into bags, four equally spaced points from the meal mix and 10 equally spaced points from the granulated microbial product feed (i.e., 1, 3, 5, ..., 35, 37 and 39 bags).

Within one treatment, uninfected diets were used in alternating soil cages and sampled at weeks 1 and 4; from the other cages at weeks 2 and 6 of the feeding experiment.

An equal number of male and female animals were slaughtered to determine the weight of each breast meat, body and small intestine. For each bird, the ratio of breast weight to gut weight and gut length is calculated.

All data are subjected to split-plot variance analysis, the differences being determined by contrast and estimation of the desired effects.

Sixty animals per treatment were taken to a university for organoleptic evaluation.

The microbial product for direct feeding, independently of the processing, increased (P <.05) the nutrient conversion compared to the corresponding control, while compared to the control

increased weight gain (P <.05) occurred only in the meal mix diet (Table 4). The microbial product used as direct food, P, increased (P> .05) the nutrient conversion versus Stafac 10, which was similar (P> .05) to the control, P.

The product had the desired level of borehole composition (Table 5).

The microbial product for direct feeding was evenly distributed in the feed. The direct-feeding microbial product, M, was at the desired level, while the direct-feeding microbial product, P, was 1 to 1.5 orders of magnitude higher than that required for starter and growth (Table 6). The high level of microbial product P for direct feeding was due to the product being too concentrated to ensure proper recovery of the microorganisms after granulation.

The numbers of soil cage samples obtained with the microbial product for direct feeding of P were in close agreement with the numbers obtained from the mixing tests (Table 7). However, the level of microbial feed suitable for direct feeding of M was reduced by 2 orders of magnitude in the growth and withdrawal mix at weeks 4 and 6.

The microbial formulation for direct feeding of M increased both breast weight and recovery (P <.05) versus the control for M (Table 8), while the microbial product P for direct nutrition showed enhancement over the control P (P>). 05). The improvement in the meal mix was in line with previous studies. THE ······"· · ·· • " · · " " • · · · ·

-20 direct-feeding microbial product P did not show a similar magnitude improvement in breast weight compared to the direct-feeding microbial product M. This failure is due to the better use of energy through granulation, which has left less room for improvement.

Granulation increased the average body weight of the birds by 96 grams compared to the meal mixture The microbial product for direct feeding increased the uniformity of the birds' weight (Figure 5), with the greatest improvement in the meal mixture.

Granulation increased the average breast weight by 15 grams in the meal mix. The microbial product used as a direct food increased the average breast weight and its uniformity (Figure 4) as compared to the control, with the greatest improvement being seen in the meal mix. Stafac showed the greatest improvement in granular feeds.

Granulation increased the amount of breasts by 53 percent for meal mixes. Microbial product M, used as a direct food, showed an 84 percent increase over M control, which is similar in magnitude to the granulation response.

The use of a microbial product used as a direct food resulted in a shorter (P> .05) small intestine than either the controls or Stafac at current lengths, or in terms of body weight or breast weight (Table 9). In the case of microbial products for direct feeding of M, the weight of the small intestine is less than that of the M control if either in actual weight, or in percentage of body weight

head it out. The loss of small bowel weight and length in the microbial product used as direct food suggests that less energy is needed for maintenance and more energy is available for growth, which is also indicated by improved food utilization and breast weight gain (Figs. 7-8).

spreadsheet)

Adding the microbial product P as a direct feed to the birds did not change the odor of the feed when Stafac

Compared with 10 (Table 10) In the second study, the microbial product used as direct food P was shown to improve thigh / leg aroma compared to P control. This improvement in aroma was not observed in the first experiment.

Table 2

Cage Allocation

Treatment

Cages serial number

control,

2,6,15,17,22,26,104,109,113,117,122,126

Direct feeding. alk.

P microbes.

product

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

Stafac 10

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

M control

3,9,13,20,24,30,102,108,114,119,121,127

Direct Feed alk.

M microbes. product

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

Cage size: 1.2 x 4.6 m, single tube feeding, one hanging

-22pots, pine shavings for maintaining cleanliness, cooling system, and well insulated, artificial heating, hanging sidewalls.

evaporator hot air • ··· »···

-23 · «♦

Table 3

The composition of the basic diets

additives Production phases departure Growth! withdrawal Ground corn 65.37 % 67.89 74.29 Weave aloe 25.58 23.53 17.83 Meat and bone meal 3.00 3.00 3.00 Fat 3.36 3, 32 2.59 De-fluorinated phosphate 0.95 0.79 0.73 Calcium carbonate 0.61 0.62 0.63 Salt 0.35 0.31 0.32 trace elements 0.05 0.05 0.05 methionine 0.39 0.28 0.33 lysine 0.19 0.06 0, 18 vitamin Mixture 0.05 0.05 0, 05

• · ·· »« «« • · «

Table 4

Production data in ground cages

control Granulate Dara mix Bowl. s.* P Stafac 10 control Bowl. s.* M Weight (kg) 2.17 ^a 2.18 ^a 2.17 ^a 2.06 ^b 2.12 ^a feed utilization 1,871 ^b 1,827 ^a 1,855 ^ab 1,917 ^c 1,856 ^ab Adj. táphaszn. . ¹ l, 832 ^b 1,789 ^a 1,807 ^ab 1,887 ^c 1,812 ^ab Weight corr táphaszn. ² 1,801 ^b 1, 755 ^a 1,775 ^ab 1,897 ^c 1,789 ^b Mortality,% 4.40 4.64 5.95 3.33 5.60

¹ Corr. táphaszn. = Total Nutrition / (live + dead weight) ² Weight corr. táphaszn. = Corr. feed - ((weight-4.60) / 6).

^abc P <.0 5 * bowl. No.: according to the invention.

-25 ··

Table 5

Product quality control and quality assurance

treatments ME ^ · number cfu / g MB ^ number product strain ratio SF202: SF301 Direct feeding. alk. P microbial product 5.75X10 ⁸ Ι, ΟΙχΙΟ ⁸ 50:50 Direct feeding. alk. M microbial product 9.64X10 ^{7 7} 9,60xl0 57:43

1. Quality control

2. Quality Assurance

-26 • ·

Table 6

Nutrition mixing test and recovery

Production Phases and Treatments Groats Mixture Granules Extraction ¹ cfu / g feed% groats

Departures

Control, P NA ² l, 06x10 ³ Directly. Tinder. P Microbes. produce. 2,02x10® 1.67X10 ⁶ 98.69 Stafac 10 SO 6,46x10® - Control, M 2.51X10 ³ Directly. Tinder. M Microbes. produce. 1,34x10®

growth

Control, P SO ² 4,86xl0 Directly. Tinder. P Microbes. produce. 3,89x10® 1,09x10® 91.62 Stafac 10 5.25X10 ⁴ 6.42x10 ³ - Control, M 1, OOxlO ² Directly. Tinder. M Microbes. produce. 1,48x10®

-276. Table (cont.)

Nutrition mixing test and recovery

Production Phases and Treatments Groats Mixture Granules Extraction ⁴ cfu / g feed% groats

Withdrawal 1, 11xlO ³ - Control, P ² 8,50xl0 Directly. Tinder. P Microbes. produce. 7.04x10 ⁴ 4.9x10 ¹⁰ 117.40 Stafac 10 8.80x10 ³ 1.79x10 ⁴ Control, M ² 8,92xl0 Direct Feed M Microbes. produce. 1.33X10 ⁵ Average Control, P 8.50X10 ² 8.28x10 ² - Directly. Tinder. P Microbes. produce. 8.21X10 ⁵ 9.64x10 ⁶ 118.09 Stafac 10 2.15X10 ⁴ 9.05-10 ³ - Control, M ² 8,72xl0 Directly. Tinder. M Microbes. produce. ⁵ 1,38xl0 ¹ Extraction log ^ Q transformed data calculated

NA meaning no data

7. Table

-28Quality assurance of soil cages

treatments 1 weeks Average 2 cl 4 : u / g feed 6 control ² 3,78xl0 ² 3,83xl0 ⁷ 8,60xl0 2.21X10 ² 4.08x10 ² Directly. Tinder. P Microbes. produce. 9.23-10 ⁵ 9.37X10 ⁵ 8.77X10 ⁵ 8.48X10 ⁵ 8.96X10 ⁵ Stafac 10 ² 8,73xl0 1.29x10 ^{2 2} 6,46xl0 ² 8,63xl0 ² 8,89xl0 Control, M ² 3,46xl0 1.26 x 10 ² 2.79x10 ³ 2, OOxlO ² 5.08x10 ² Direct Feed M Microbes. produce. 1.43Χ10 ⁵ 1.25X10 ⁵ l, 75x10 ³ 1, OOxlO ^{3 4} 2,32xl0

Table 8

Breast weight evaluation

control Granulate Dara mix Bowl. s.* P Stafac 10 control Bowl. s. M Weight, g 2240.7 2230.1 2195.9 2143.8 2149.9 Breast weight, g Breast weight a 234.4 ^a 239.6 ^a 232.0 213.3 ^b 229.6 ^a % body weight ^ab P <.0 5 * Bowl. s. = a 10.51 is ^the invention 10.68 ^a 10, 58 ^a 9.93 ^b 10.67 ^a

Table 9

Gut weight and gut length

control Granulate Dara mix Bowl. s.* P Stafac 10 control Bowl. s.* M Weight, g 2240.7 2230.1 2195.8 2143.8 2149.9 Breast weight, g 234.4 ^a 239.6 ^a 232.0 213.3 ^b 229.6 ^a WB weight, g 92.6 93.3 93.4 91.4 87.4 VB length cm 196.75 191.26 194.54 193.29 191.26 VB, g / cm 0.48 0.48 0.48 0.47 0.46 VB weight, g / 100g body weight 4.17 4.18 4.27 4.29 4.08

VB length, cm / 100g

body weight 8.81 8.64 8.97 9, 17 8.97 VB weight, g / lOOg breast weight 40.19 39.70 40.97 43.96 38.69 VB length, cm / lOOg breast weight 84.86 ^a 81.97 ^a 85.65 ^a 93.70 ^b 84.86

ab

P <. 05

Bowl. s. * = according to the invention

VB = small intestine

-30 ··························································································································································ing ·

Table 10

Taste panel evaluation

Group The number of correct identifications is ¹ Fabric comparison First test Trial 2 Combined Thigh / leg Stafac 10 / Inspection, P 6 3 9 Stafac 10 / XINOC, P 3 4 7 Directly. Tinder. microbes P / Control P 2 8 * 10 Breast Stafac 10 / Inspection, P 2 6 8 Stafac 10 / XINOC, P 1 3 4 Directly. Tinder. microbes p / Control P 5 4 9 * THE evaluators are capable they were statistically j opposite number (P <.05) unusual patterns

¹ The number of correct identification of the unusual sample at the 5% significance level was 7 for n = 10 and 11 for n = 20.

Broiler studies were conducted to determine the efficacy of a direct-fed microbial product in meal mix and granular feeds. The microbial product suitable for direct feeding, irrespective of processing, only improved (P <.05) the nutrient utilization compared to the appropriate control. Direct feeding P microbial product improved nutritional utilization (P> .05) versus Stafac 10, which was similar to P control (P> .05). Microbial product M, which is directly fed, increased both breast weight and yield compared to Control M (P> .05), while microbial product P, used as direct food, showed improvement (P> .05) as compared to Control P . Birds treated with P microbial product for direct feeding did not contain more flavor than Stafac 10 treated

Example 6

One hundred and forty, four mixed piglets (average starting weight kg) were randomly distributed by sex and weight into sheds on slate-covered soil (Table 11) and fed for 119 days. Table 12 shows the composition of the basal growing and finishing feeds. Growth rations were applied until the average weight in each cage was 54.5 kg, followed by the final rations and slaughter. Each feed contained Mecadox (50 g / t), up to a weight of 34 kg for pigs, and 100 g / t of chlorothetracycline up to 54.5 kg. The experiment consisted of a negative control and a feed containing a selected microsphere direct feeding product containing the microbial product at a concentration of 1x10 ⁴ CFU / g feed. All feeds were used in the form of a meal mix. Six replicate cages were used for each experimental feed, each with 12 brood mills.

Upon arrival at the experimental site, the mixed brood pigs were given Ivomec to expel external and internal parasites. Four weeks later they were given Safeguard to drive out the whipworms.

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Body weights, nutritional utilization and mortality were recorded for each cage. The nutrient conversion was calculated for each cage

Prior to assay, the microsphere culture concentrate was diluted with calcium carbonate. Theoretical product CFU was 2x10 ⁷ CFU / g product. To determine the current CFU number, an 11 gram sample of the product was tested. The sample is plated on standard microwell bacteria as described for standard lactic acid bacteria.

An additional one gram sample of the product was tested in duplicate to confirm the CFU number and strain composition of the product.

Each treatment point was sampled weekly and assayed for the microspheres in the same manner as described for lactic acid bacteria.

The product was found to contain the desired concentration of the desired microorganism (Example 14).

The recovered microorganisms ketrecmintákból sama varied between ¹ and lxlO- 1,6x10® CFU / g of feed (Table 15). The two extremes are attributed to the sampling or plotting error. The rest of the samples were around the target lx10 ⁴ CFU / g feed level.

The microsphere-encapsulated product increased (P> .05) body weight gain and nutritional conversion over 28 days (Table 13). Pigs were hit by a TGE outbreak in the first week of the experiment. This may have been one of the reasons why there was a 28-day incubation period before a response was observed, and another reason for the pigs' digestive

-33 had to adjust to the product. It can be seen that the microspheres of the microbial product suitable for direct feeding according to the invention are effective also in pigs, chickens and turkeys.

From the examples above, it can be seen that the process of the present invention achieves its objects

Table 11

Selection of cages

Experiment number: 670-9102

treatments Numbers cage control 3, 4, 6, 9, 11, 12 microbial product 1, 2, 5, 7, 8, 10 Cage Size: 3.6 x 14.6 m, a four-hole Smidley feeder, egycsecses driker for temperature control sprayers we used it partially slate

covered floor, modified open front building

Table 12

The composition of the basic rations

Experiment number: 670-9102

additives production phases Grow closing Ground corn 76.30 82.20 Weave aloe 21.25 15, 50 Dicalcium phosphate 1.05 0.90 Calcium carbonate 0.85 0.85 Salt 0.30 0, 30 Vitamin / trace element premix 0.25 0.25

Table 13

Soil cage production data

Experiment number: 670-9102

control

Microbial product

Day 14

Weight Growth, kg 4.35 4.17 feed conversion 2,439 2.483 Mortality% 1.41 1.39 Sun Weight Growth, kg 11.93 12.66 feed conversion 2,405 2,212

Mortality% 2.82 1.39 42nd Sun Weight gain, kg 20.77 21,23 feed conversion 2,497 2,428 Mortality% 2.82 2.78 56th Sun Weight gain, kg 31.34 32.43 feed conversion 2.507 2.457 Mortality% 4.23 2.78 70. Sun Weight gain, kg 40.55 41.28 feed conversion 2.735 2,674 Mortality% 5.63 2.78 84th Sun Weight gain, kg 50.35 50.80 feed conversion 2,904 2,882 Mortality% 5.63 2.78 98th Sun Weight gain, kg 58.92 61.10 feed conversion 3,071 2.988 Mortality% 5.63 2.78 112 . Sun Weight gain, kg 69.04 70.17 feed conversion 3,164 3, 134 Mortality% 5.63 2.78 119 . Sun Weight gain, kg 73.66 75.11 feed conversion 3.217 3, 177

···· ····

Mortality%

5.63

4.17

Table 14

Product quality control and quality assurance

Experiment number: 670-9102

Product ME number MB number Stock ratio

CFU / g product SF202: SF301

Microbial product

4.3 x 10 ⁷

Table 15

Soil cage quality assurance

Experiment number: 670-9102

Date control 1991st 5:03. 1.7x10 ³ 1991st 5:08. 1.8x10 ⁴ 1991st 5:22. 0 1991st 5:30. 0 1991st 6:05. 2, Ox ¹⁰ 1991st 6:12. 1, Ox ¹⁰ 1991st 6:19. 0 1991st 6:26. 6.7x10 ² 1991st 7:03. 4.9x10 ⁴ 1991st 7:10. 5.2x1ο ¹

Microbial product

7.4x10 ³

1,6x10®

1, Ox ¹⁰

2.6x10 ³

2.2x10 ⁴

2, OxlO ⁴

9.6x10 ³

5.6x10 ³

3.2x10 ³

3, Ox ¹⁰

1991st 07.17. 1 OxlO ² 4.5x10 ³ 1991st 7:24. 0 1, OxlO ⁴ 1991st 7:31. 0 1.2x10 ⁴ 1991st 8:07. 0 6.4x10 ⁴ 1991st 8:14. 0 7.7x10 ³ 1991st 8:21. 0 1.2x10 ⁴ 1991st 8:28. 0 9, lx10 ³ 1991st 9:04. 0 2.9x10 ⁴ 1991st 9:18. 0 8.9x10 ³ 1991st 9:25. 0 5.5x10 ³ Average 9 5x10 ° 8.4x10 ³

Claims (13)

CLAIMS 1. A method of increasing the growth of an animal comprising administering to the normal animal feed a small amount of a microbial product having direct growth effect which can be used for growth and which is essentially Enterococcus faecium 301, ATCC No. , viable, stable, microspheres of dried fatty acid, and Enterococcus faecium 202, ATCC No. contains microspheres of fatty acids. Process according to claim 1, characterized in that the fatty acid microspheres are produced on a rotary disk. 3. A The method of claim 2, wherein the microbial product for direct nutrition is about It contains 30-70% of one type of fatty acid microsphere, while the rest the other kind z 4th Claim 3 fatty acid ^c 12 ^_c 24 free 5th Claim 4 fatty acid stearic acid. 6th Claim 1 fatty acid. microsphere of fatty acid. The method of said method, characterized in that the method is characterized in that it is present in about the same amount. 7. The method of claim 1, wherein the amount of microbial product used as direct feed to the feed ration is about 0.014-0.907 kg / tonne feed. The method of claim 7, wherein the amount of microbial product used as direct feed is about 0.362-0.544 kg / tonne feed. 9. The method of claim 7, wherein the microbial product for direct feeding is between 1x10 ⁵ CFU / g and 2x10 ⁸ CFU / g. The method of claim 9, wherein the microbial product for direct nutrition has a microorganism number of about 1 x ^{10 8} CFU / gram. 11. The method of claim 1, wherein the animal is a chicken. 12. The method of claim 1, wherein the animal is a pig. 13. A microbial product for direct nutritional growth enhancement in animals comprising essentially microspheres of viable, stable, dried fatty acid Enterococcus faecium 301 and microspheres of viable, stable fatty acid Enterococcus faecium 202. The microbial product for direct nutrition according to claim 13, characterized in that it contains from about 30% to about 70% of one of said streptococci and the remainder contains the other streptococcus. Suitable for direct feeding 15. The microbial product of claim 14, wherein the fatty acid ^C i2 ^-C 24 free fatty acid. 16. The microbial product for direct nutrition according to claim 15, wherein the free fatty acid is stearic acid. 17. A microbial product for direct feeding according to claim 16, characterized in that it comprises a -40streptococci are found in almost equal amounts. 18th The composition of claim 13, wherein well analyzed, that the animal chicken. 19th The composition of claim 13, wherein well analyzed, that the animal pig.