SI9300128A - Fatty acids microspheres containing enterococcus for use to enhance growth and improve environment 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 ACIDS INCLUDING MICROSPHERE TO PROMOTE GROWTH AND IMPROVE QUALITY

BACKGROUND OF THE INVENTION

Antibiotic growth promoters are used for poultry, namely chickens and turkeys. Growth promoters such as Stafac ^R and BMD ^R (bacitracin methylene dishalicylicate) are known antibiotics and are used at sub-therapeutic levels, e.g., 10 grams per tonne and 25 grams per tonne as food additives, to achieve the desired poultry growth characteristics. Recently, however, the use of antibiotics for these purposes has become a target of criticism. One criticism is the possibility that poultry may eventually develop antibiotic tolerance and antibiotics may no longer work well to promote growth. Other concerns relate to health due to unnatural antibiotic supplements and the negative impacts they could cause. However, due to the benefits of antibiotics, these are still commonly used to improve food, skeletal composition and promote growth.

Certain bacteria are known to be potentially useful when added to animal feed. These bacteria are useful in supplying naturally occurring intestinal micro-flora. Some companies sell microbes for direct use in a diet containing the bacteria they want. Microbes for direct nutritional use, however, have some difficulties in maintaining a stable product. Usually, microbes for direct use in the diet are used at fairly low doses added to food at dosage 0, IX. However, farmers often store unused food containing germs or nutritional supplements for long periods of time. Storage conditions are often high temperature and some humidity. In many cases, the humidity is just enough for the bacteria to activate and start to grow, but the amount of moisture is not large enough for the bacteria to survive. That's why he died. So the activity of microbes for direct use in the diet is stopped. In other cases, the addition of antibiotics to the microbes for direct use in the diet or in dietary supplements has the opposite effect on the bacteria, especially if less moisture is present and the bacteria die again. So there is a serious problem of microbial stability for direct use in the diet, during storage for extended periods of time.

In other environments where microbes are used as a supplement for direct nutritional use, e.g. when feeding chickens, usually the material with the addition of microbes for direct use in the diet is encapsulated. The moisture from the steam used during encapsulation partially activates the bacteria, but can also kill them due to insufficient moisture content. They can also be killed by fever during encapsulation. There is also the problem of the acidic environment of the stomach, which can deprive the bacterium of all its effectiveness before it reaches the gut. Therefore, there is a continuing need for direct microbes for nutrition that will release organisms in time to the intestine without prior release due to moisture or adverse pH conditions existing in the digestive tract before the small intestine.

It is desirable to achieve, as far as possible, certain characteristics in poultry. These are increased weight, better food conversion, skeletal composition, and ultimately uniformity of the flock.

Increased weight and better food conversion are, of course, desirable

Fzaradi accompanying economy to accompany these desired results. The composition of the skeleton is important because the most desirable area is suitable for storing tissue, the breast of animals, so that we can offer large quantities of selected meat. So it's not just the weight that matters, because where the weight is, the skeleton is just as important. Harmonizing the weight of the flock is important, because more birds, when they are normal size, less work is needed and processors can rely much more on machining. On the other hand, if the weight of the birds varies from very small to very large and the total weight of the flock may even be the same, small and large birds require a lot of manual labor because of their deviation from the average size. So the weight equivalence in the flock, with as many percentages of normal-sized birds as possible, so that the chickens can be processed on standard facilities is a very desirable feature.

Similarly, microbes for direct use in nutrition that would not be suitable only for poultry such as e.g. chicken and turkey but would also be useful in pigs.

The primary object of the present invention is to provide poultry germs for direct use in diets containing no antibiotics and containing only fatty acid microspheres containing natural organisms.

Another primary object of the present invention is to provide microbes for direct use in a diet containing two organisms, namely Enterococcus Faecium 301, DSM No. 2. DSM-Nr. 4789 and Enterococcus Faecium 202, DSM-No. DSM-Nr. 478S. DSM is a collection of bacterial cultures in Germany. DSM stands for Deutsche Sammlung von Microorganism located in the city

Braunschweig in West Germany. These organisms will be deposited with the ATCC, with all restrictions on admissible claims.

It is further an object of the present invention to provide microbes for direct use in the diet that allow poultry to gain weight, allow for better conversion of food, allow greater supply of breast meat, and which ensure uniformity of flock weight in the normal weight range.

Another primary object of the present invention is to provide microbes for direct use in a diet suitable for supplementation to a poultry diet containing microsphere-shaped bacteria using a special rotating disc technique using free fatty acid matrices.

A further object of the present invention is to provide microbes for direct use in the diet that have a stability in the range of 3 to 6 months without noticeably reducing the number of organisms.

It is another object of the present invention to provide a process for forming spheres by a rotary process containing dried bacteria, ensuring that they have the same size.

Another object of the present invention is to provide, by means of a rotating disc, spheres of dried, free-flowing bacteria that can be easily inserted into doses of poultry food.

It is further the object of the present invention to provide microspheres of a fatty acid material containing certain bacteria that are useful to the sphere as germs for direct use in the poultry and prey foods.

BRIEF DESCRIPTION OF THE DRAWINGS

Figures 1, 2 and 3 graphically illustrate the stability of the genera using the stearic acid matrix.

Figure 4 is a diagram showing the distribution of a breast yield upon experimental feeding of a microbial for direct nutritional use of the present invention.

Figure 5 is a diagram showing the distribution of body weight when fed experimentally with an ingredient from microbes for direct use in the diet of the present invention.

Figures 4 and 5 show controls using antibiotics and using microbes for direct use in the diet of the present invention.

DESCRIPTION OF THE INVENTION

The invention is a method and composition for promoting growth for poultry and pigs, which, when added to normal dietary doses, contains a small but growth-promoting amount of microbes for direct use in the diet containing dried microspheres of fatty acids Enterococcus Faecium 301, DSM No. DSM-Nr. 4789 and the dried fatty acid microspheres of Enterococcus Faecium 202, DSM No. DSM-Nr. 4788, where it is desired that the fatty acid microspheres be formed by drying on a rotating disk.

DETAILED DESCRIPTION OF THE INVENTION

It has been surprisingly discovered that promoting the growth of poultry and pigs can be achieved by adding normal dietary doses and certain amounts of Enterococcus Faecium 301, DSM No. 1 fatty acid microspheres. DSM-Nr. 4789 and certain amounts of Enterococcus Faecium 202 fatty acid microspheres, DSM No. 2 DSM-Nr. 4788. The fatty acid used may be any of Cia> to free fatty acids, and stearic acid is preferred. The organisms are preferably present in about the same doses, but the dose may range from about 30Χ to about 70Χ of one organism. the rest is occupied by another organism.

It is not known exactly why these two organisms produce the desired results of the present invention, especially for poultry, namely weight gain, better food conversion, improved breast yield and increased uniformity of birds within the flock. The fact is that they work on the condition that both are used in combination so that they can interact with one another in some way and provided that they are used within the limits stated above. These combinations are one feature that somehow interacts and cooperates to produce the desired results of the present invention, resulting in significant improvement in poultry bones, meat quality and processing. Similar results can be obtained for pigs, as indicated in the examples,

The dose of microbes for direct food use in the food sector can fluctuate considerably but will generally be in the range of approx. 0.226795 kg to approx. 0.90718 kg per tonne of feed, mainly from approx. 0.362872 kg to approx. 0,544308 kg and usually approx.

0.45359 kg per tonne of feed. The number of organisms, that is, the number of colony forming units (CFU) per gram present in microbes for direct dietary use, can also fluctuate within the range of approx. 1 x 10 “* CFU / gm up to approx. 2 x 10 * ”CFU / gm, preferably about 2 χ 10® CFU / gm.

When microbes for direct nutritional use, as described above, are added arbitrarily to animal rations, the combination of the two genera of the organisms mentioned here behaves as a growth promoter. Growth enhancers in use now contain antibiotics such as Stafac ^R and BMD. The advantages of the sub-therapeutic level of antibiotics, as growth enhancing additives, can be achieved by naturally occurring organisms of the present invention that allow microbes for direct dietary use to be made in accordance with the present invention and added according to the method described herein. In fact, there have been some experiments suggesting that the combination of microbes for direct use in the diet and growth promoters together outweighs the benefits of each individually, so they can be used together if necessary. However, in most cases it is desirable to use the microbes for direct use in the diet itself, since one of the objects of the present invention is to completely avoid the use of growth promoters.

The method of treating organisms is not critical, as long as the organisms can be kept alive, given to animals and placed in such a way that they will mix well with animal food, and are usually uniform in form so that dosage can be controlled.

The desired ways of achieving these requirements are to ensure that microorganisms are located in the microsynthesis of the fatty acid matrix. Microsphere means a matrix of fatty acids into which a multitude of organisms are incorporated. It differs from the microcapsule where individual organisms are encapsulated. In the microsphere, the fatty acid matrix acts on the composition similar to the ratio of the biscuit matrix from the dough to the chocolate chip, where the chips represent a group of organisms. This process is described in the original application by co-inventor Rutheford and others. This process binds the bacteria to the heated fatty acid. The temperature of the fatty acid and the exposure time of the bacteria to the fatty acid are controlled to keep the bacteria alive and to allow mixing with the fatty acid. The mixture is placed on a rotating disk and the result is a microsphere of bacteria with fatty acid that acts as an array. Many important benefits are achieved through this process. First, the bacteria remain alive throughout the process; second, the process co / nbined by the rotating disc technique allows uniform sizes of microspheres for improved dosage. Third, the nature of matrices, fatty acid, permits the formation of unique microspheres. The combination of these factors gives us highly stable microbes for direct use in the diet, with maximum efficiency.

In the original application process, it is important that the microspheres are formed where each sphere contains a plurality of bacteria in a free fatty acid matrix rather than a single microcapsulator of each bacterium individually in a coating or film-like layer of fatty acid. This provides stability benefits and more effective dosage in bacterial treatment.

The desired matrix agents are C ± _a to C ₂ a free fatty acids. Since a free fatty acid mixture could be used, it is desirable to use a single pure free fatty acid. It is also desirable that free fatty acid is unsaturated fatty acid, most preferably stearic acid.

It is generally important that the fatty acid has a melting point below 75 ° C, preferably in the range of 40 ° C to 75 ° C. Of course, at room temperature, it must be solid to be an effective matrix. All free fatty acids that fall within the limits of the chemical description given here meet these requirements.

To increase the stability of the product, the bacteria that are added to the product are usually freeze-dried bacteria. So we can revive them by adding moisture.

Made in the microsphere according to the procedure described below, the microsphere typically contains from approx. 50% to over 90% by weight of the fatty acid component and the rest is bacterial culture. The desired range is approx. 60% to 75% fatty acid. If too little fatty acid is used, the matrix will be unsuitable for protection. On the other hand, if used too much, the matrix will be too thick, resulting in untimely release in the gut.

The method used in the present invention is a method for forming microspheres by means of a rotating disk. In general, the sludge from the rotating disc technology, bacteria and fatty acid components is thoroughly mixed and the mixture is added in single doses to the center of the stainless steel rotating disc. From the center, the mixture is thrown outward as a result of centrifugal force and forms microspheres. It is then collected in a cooling chamber where atmospheric or slightly lower conditions are maintained, weighed and prepared for packaging.

While encapsulation per se by rotating disk is known, the production of microspheres contained in arrays without shell is not known, nor is the use of the microspheres process or encapsulation by freeze dried bacteria known.

For a description of encapsulation using a rotating disk, see a document published by Johnson et.al. from the Southwest Research Institute of San Antonio, in Journal of Sas Chromotoqarphy, October 1965, pages 345-347. In addition, the rotary disc machine is described in detail in United States Letters Patent, Sparks, 4,675,140, issued June 23, 1987, under the title Method for Coating Particles For Liguid Droplets, the disclosure of which is stated herein. Anyway, the most desirable process is the one described in the original application.

It is important to note that the production of microspheres by the rotating disk process is significantly different from that obtained by conventional tower spray drying or microencapsulation. In the conventional tower spray drying process, there is a tendency for fragments to clump together, which is very poor for the coating, as well as for the stability of the product, for significant damage, perhaps from a few days to a few weeks. This process provides a shell coating around the object and the bacteria have proved to be too small to retain or provide a uniform size that would be practically usable. By forming microspheres, in part with the agents used in the present invention, the stability of the resulting bacterium, even when slightly exposed to antibiotics and moisture, is from three to six months, while maintaining the ability to live in equally distributed particles.

When the free fatty acid microspheres of the present invention are used within the limits of the present invention, the disk, which is typically 101.6 mm to 152.4 mm in diameter, rotates at a speed of 2000 rpm / minute to 4000 rpm, most preferably approx. 2500 rpm to 3200 rpm, with supplemental doses of 50 g to 200 g per minute. Currently Known Wanted! conditions are the use of stearic acid, the use of two organisms described above, a disk of 101.6 mm diameter rotating at 3000 rpm and a supplemental dose of 100 g per minute of bacterial stool and stearic acid containing 35% bacteria and 65% stearic acids. When this is done, a product whose particle size is from 75 microns to 300 microns is obtained, and the desired size is less than 250 microns.

The following examples are available, for illustration, but not limitation, of the process of the present invention. Some examples are described in connection with Figures 1, 2 and 3. Examples 1 to 4 and Figures 1, 2 and 3 are in connection with my previous invention. Example 5 and Tables 2 to 10 refer to the process of the present invention for poultry microbes for direct use in the diet. Case 6 relates specifically to turkeys and case 7 relates to pigs.

Example 1

Example 1 is related to Figure 1. It shows the product stability of two different genera of Enterococcus feacium at 4 ° C and 27 ° C. As shown in Figure 1, the stability of the encapsulated genera of the Enterococcus feacium product encapsulated by a rotating disc is observed using stearic acid at a dose of 35% by weight of culture.

The microsphere forming conditions were as described above, namely, 35/65 bacterial / stearic acid sludge at 60 ° C, and a diameter disc was used

101.6 mm, rotating at 3000 rpm, with a supplemental dose of 100 g per minute. Spheres were formed, placed in hot sealed bags, and adverse impacts were recorded weekly to determine CFU. It is apparent that the product of the present invention very well maintained the values of the CFU unit for colony formation of the organism up to a storage time of 70 days.

Example 2

Example 2 should be interpreted in conjunction with Figure 2. The figure shows the stability of the individual encapsulated species mixed in the normal feeding ratio in the presence of three poultry antibiotics. The dose consisted of the following ingredients:

54% finely ground cereals

26% soybean meal

2% fish coarse ground flour

1.5% dicalcium phosphate

1% limestone

5.5% soybean oil

12% moisture content

Three antibiotics were added at the following mass doses:

- Decoquinoate 6% (454 ppm)

- Salinomycin (50ppm)

- Sodium monensin (120 ppm)

The culture was added to this mixture at such a dose that approximately Ιχΐθ * ⁶ · CFU / gm of feed was obtained. Feed was packed in hot sealed bags and incubated at room temperature Samples were taken weekly to determine CFU. The graph in the picture shows us excellent stability.

Example 3

Example 3 should be interpreted in conjunction with Figure 3, which shows the stability of an encapsulated feed of Enterococcus faecium in feed in the presence of various antibiotics. The dose consists of 60% finely divided corn, 38Χ soybean meal and 27 limestone with a moisture content of approx. 14Χ. The culture was added to a level of about 10 ^ CFU / gm and mixed. The 4.54 kg quotas were stored in airtight bags at 20 ° C, and samples were taken weekly for 16 weeks. Antibiotics were included in the following doses:

Bacitracin methylene disalicylate 50 gm / ton Carbadox 50 gm / ton Chlorotetracycline 200 gm / ton Lasalocid 30 gm / ton Lincomycin 100 gm / ton Neomycin 140 gm / ton Oxytetracycline 150 gm / ton Sulfamethazine 100 gm / ton Tylosin 100 gm / ton Virginiamycin 20 gm / ton ASP250 100 gm / ton Furadox 10 gm / ton

Table 1 is a list of minimum times during which one colony formation unit (CFU) is lost.

Table 1

Time in the days of porridge

Antibiotic for loss of one CFU unit at 20 ° C and 14% moisture

Storage time

In days!

Control 103 Bacitracin 88 CarbadoM 54 Chlortetracycline 60 Lasalocid 57 Lincomycin 75 Neomycin 53 Oxytetracyc1ine 59 Sulfamethazine 62 Tylosin 52 Virginiamycin 112 ASP250 67 Furadox 53

Example 4

In Example 4, the stability of the product for use in chicken feed was determined after encapsulation. The microencapsulation conditions were as described above. The conditions used in this study were as follows:

Crude protein, not less than crude fat, not less than crude fiber, not more than

18.07.

5.07.

6.07.

Pills with and without antibiotics (CTC 50 mg / tonne) were made with the following ingredients and under the following conditions.

Cereals, SBM, whey, soybean oil, dicalcium phosphate, limestone, rock mineral premix, vitamin premix, selenium, copper sulfate. The culture was added at about 5x10 ”CFU / gm feed (100-150 gm / tonne).

The air-conditioning temperature was 70 ° C and the non-colored capsules had 78 ° C.

The capsules were stored in non-airtight bags and sampled weekly for CFU determination.

In all cases, the encapsulated products were not damaged by the conditions of encapsulation (pelletization). For some, the pelletized product showed some stability rather than the pelletized product.

Example 5

Four thousand five hundred sixty, Peterson x Arbor Acres broilers, one day old chickens were randomly selected and placed in floor enclosures (Table 2) with refurbished litter and kept for 45 days. All birds that died within the first five days were replaced by a bird of the same sex from the same consignment and treatment. The composition of the basic diet at the beginning, during the growth and the finished diet, and the relationships are shown in Table 3. The initial diet, the growth diet and the finished diet were formulated to contain 1425, 1450 and 1475 kcal ME / lb and each individually 90 g / tonne of monsein »Initial doses were fed at 1 to 21 days of age, fed with growth during growth, fed between 21 and 42 days, and finished diet between 42 and 49 days of age. Negative controls were discussed, {Control, M)? selected encapsulated microbial cultures for direct use in the diet containing Enterococcus taiga j um 301, DSM No. DSM-Nr. 4789 and EnterPCPCCUS faeeium 202, DSM No. DSM-Nr.4788 each encapsulated in a fatty acid by means of a rotating disc as described in Example 1 and each present at a dose of 50% of microbes used for direct use in a 1 x 10 diet ^with CEU / g diet, porridge (microbes for direct use in diet M); negative controls, pelleted (Control, P); microbes for direct food use zix 10 * · CFU / g pulp, pelleted (microbes for direct food use, P); and positive controls used at 10 g / tonne virginiamycin pelleted (Stafac. Initial feed ration was crushed for pelletized treatments. Twelve equal enclosures with 35 male and 35 female animals were used with each experimental meal .

Body weight, food intake and mortality in the first five days were recorded across the fences. For each enclosure, food conversion, adjusted food conversion, and body weight with adjusted food conversion were calculated.

All data were subjected to non-compliance analysis and differences were detected using Fisher LSD.

Prior to the study, the culture of microbes for direct dietary use was expanded with calcium carbonate. Theoretically

1ώ the number of microbes for direct use in the diet, M and microbes for direct use in the diet, F 'were 1 x 10® and 2 x 10 ^ CFU / g of product, respectively. A sample of the leagues of each product was individually tested in duplicate to determine the actual number of the product. Each sample was plated using a standard plating technique for encapsulated lactic acid bacteria.

A mixing test was performed for each production phase. The test was designed to ensure that microbes for direct dietary use were uniformly distributed at appropriate doses in food and survived pelletization. Each dose was tested at the time of packing in bags containing four equally spaced samples for the treatment of animal food and ten equally spaced samples for pelletizing treatment (i.e., bags 1, 3, 5, 35, 37 and 39).

We took samples of uncontaminated food alternately from half of the floor rails during Weeks 1 and 4; from the rest between weeks 2 and 6 during this study.

The same number of birds of each sex were sacrificed to determine the individual weights of the bust, body and small intestine, and the length of the small intestine. We calculated the thoracic yield and the ratio of weight to gut length for each bird.

All data were subjected to split-plot discrepancy analysis and differences were found with comparative and evaluation findings for specific impacts. Sixty birds from each treatment were sent to the university for taste evaluation using the senses.

Regardless of the process, microbes for direct nutritional use improved (P <.05) the conversion of food via individual Control with an increase (P <.05) of weight only through Control in the slurry (Table 4). Microbes for direct nutritional use, P increased (P> .05) food conversion via Stafac ”, which was similar to (P> .05) Control, P.

The product was in the desired dose and composition of the genera.

(Table 5).

Microbes for direct nutritional use were evenly distributed in food. Microbes for direct dietary use, M was at the desired dose, whereas microbes for direct dietary use, F 'i to i-i / 2, were higher than desired for doses at baseline and during growth (Table 6). The high number of microbes for direct use in the diet, P was the result of over-engineering the product to ensure sufficient replacement of organisms after pelletisaoi,

Samples of floor enclosures for microbes for direct use in the diet, P almost corresponded to the number from the mixing tests (Table 7), However, microbes for direct use in the diet, M dropped by 2 units in weeks 4 and 6 in mixtures during growth and finishing mixes.

Microbes for direct nutritional use, M improved breast weight and yield via Control, M (Table 8), while direct nutrition microbes, P showed improvement (P> .05) via Control, P. Improvement in animal the feed matches the results obtained in previous experiments. Microbes for direct dietary use, P did not show the same magnitude of improvement in breast yield compared to observed microbes for direct dietary use, M. This failure could be due to increased energy use in pelletizing, resulting in a lesser potential for improvement.

Pelletizing increased the average weight of birds by 96g via food. Direct-fed microbes increased the uniformity of bird weight (Figure 5) with the greatest improvement in animal feed.

Pelletizing increased the average breast weight by 15 g over food. Direct-eating microbes increased average breast weight and uniformity (Figure 4) via Control, the largest improvement found in food. Stafac ^R showed the greatest increase in the uniformity between pelletized

Pelletizing increased the breast yield by .53 percent units'through food. Microbes for direct nutritional use, M, showed an increase of .84 percentage points via Control, M, which was similar in magnitude to the corresponding pelleted food.

Treatment with microbes for direct nutritional use resulted in a shorter (P> .05) small intestine than any of the Controls or Stafac ^R when expressed in actual length, in proportion to either body weight or breast weight. Microbes for direct use in the diet, M were smaller in small intestine weight than Control, M expressed either as actual weight or as a percentage by body or breast weight. Reducing the weight and length of the gut to handle microbes for direct nutritional use means less energy needed for maintenance and more energy available for growth than mentioned in improved food conversion and breast yield (Table 7-8).

With microbes for direct nutritional use, P-treated birds did not lose taste compared to Stafac ^R 10 (Table 10). In the second experiment, microbes for direct dietary use, P, were found to improve leg / thigh taste compared to Control, P. This improvement in taste was not observed on the first experiment.

TABLE 2 Determination of Fences

PROCESSING

NUMBER OF FENCES

Control, P

Column ^R 10

Control, M Microbes for dir.u.,

Microbes for dir.u.,

4,724 m, one tube feeder,

Fence dimensions 1.28 m single hanging water inlet, dirt extraction, cooling source for energy source and vapor, well insulated, additional heating with hot air, side curtains.

TABLE 3

Composition of basic meals

Products phases Ingridients The beginning Growth Zak 1 j cock 7, Basic cereals 65.37 67.89 74.29 Soybean meal 25.58 23.53 17,83 Meat and bone meal 3.00 3.00 3.00 Fat 3,36 3,32 2.59 Deflourinated phosphate 0.95 0.79 0.73 Calcium carbonate 0.61 0.62 0.63 Sol 0.35 0.31 0.32 A rock mineral 0.05 0.05 0.05 Methionine 0.39 0.28 0.33 Lysine 0.19 0.06 0.18 Vitamin premix 0.05 0.05 0.18

TABLE 4

PRODUCTION DATA OSRAD

Pelletized mushy

And in The staff And in Control F ' 10 Control M Weight, kg 2,17- 2,18- 2,17- 2.06 = ³ 2,12- Conversion food 1,871 * = · 1,827- 1,855- = ³ 1,917 * = 1,856- ' Approx. BC food ¹ 1,832 = ³ 1,789- 1,907- = ³ 1,887 * = 1,812- '

Weight adjust.

conversion food ² 1,801 = ³ i, 755- 1,775- = ³ 1,897 * = 1,798 = ³ Mortality, 7. 4,40 4,64 5.95 3,33 5.60

Custom conversion food = total food / {live = dead weight)

Weight of food preparation - Food delivery - ((weight - 4.60) / 6) abc P <.05.

Inv = Invention {invention op. prev.)

TABLE 5

PRODUCT QC AND QA

Processing

QC ^X

YES ² Ratio of genera

Microbes for dir.up.P Microbes for dir.up.M

-CFU / g product- SF 202: SF301

5.75 x 10® 1.01 x 10® 50! 50

9.54 x 10 ^ 9.60 χ 10 ^ 57:43

1, Quality control

2, Quality assurance

TABLE 6

MIXING TEST AND RENEWALS

Production stages

and processing Porridge Pelletized Renewals -CFU / g of food -% porridge- The beginning Control, P NA = 1,06X10 = - Microbes for du, F ' 2.02x10 * · 1,67x10 * · 98.69 Column ^R 10 NA 6,64x10 = - Control, M 2,51x10 = Microbes for du, M 1,34x10 = Growth Control, P NA 4,86X10 = - Microbes for du, P 3.89x10 * · 1,09x10 * · 91.62 Column ”10 5,25x10 * 6.42x10 = - Control, M 1,00x10 = Microbes for du, M 1,48x10 = Conclusion Control, P 8.50x10 = 1.11X10 = - Microbes for du, P 7,04x10 * 4,91x10 = 117.40 Column ^R 10 8.80x10 = 1,79x10 * - Control, M 8.92x10 = Microbes for du, M 1,33x10 =

The way

Control, P 8.50x10 = 3,28X10 Microbes for du, P 8,21x10 = 9,64x10 Column ”10 2,15x10 * 9,05x10 Control, M 8,72x10 = Microbes for du, M 1,38x10®

Discoveries calculated on the basis of log 10 transformed data

In ways not available

TABLE 7

IT'S ABOUT FENCES

THESE DAYS

Processing

4 6

An asset

Control Mikr, for du, P Staple ”10 Control, M Mikr, for du, M

3,78x10 =

9,23x10®

8.73x10 =

3,46x10 =

1,43x10 =

3,83x10 =

9,37x10®

1,29x10 =

1,26x10 =

1,25x10®

8.60x10 =

8,77x10 »

6.46x10 =

2.79xl0 ³ l, 75x! 0 ³

2,21x10 =

8,48x10®

8.63x10 =

2,00x10 = l, 00xl0 ³

4,08x10 =

8.96x10 =

8.98x10 =

5.08x10 =

2,32x10 *

Jt,

Body weight g Breast weight g Breast yield,% body weight

TABLE 8

BREAST-BREAST VALUATION

Pelletized

Control

Porridge

Inv Stafac Inv

P 10 Control M

2240.7 2230.1 2195.9 2143.8 2149.9

234.4 · 239.6 "232.0" 213.3 ^ 229.6 "

10.51 "10.68" 10.58 "9.93 ^b 10.67" ab P. <05

Inv = Invention

TABLE 9

TE2A AND D0L2INA GUTS

Pelletized

Porridge

Inv Stafac

And in

Control P

Control M

Body weight g 2240,7 2230,1 2195,9 2143,8 2149,9 Chest weight Mr 234,4 " 239,6 " 232,0 " 213,3 * 229,6 " SI weight g 92.6 93,3 93,4 91,4 87,4 SI length cm 193,8 191,3 194,6 193,3 191,3 SI g / cm 0,476 th most common 0,484 th most common 0,480 0,472 th most common 0,417 th most common

SI weight, g / 100g body weight 4.17

SI length, cm / 100g

4.18 4.27

4.29

4.08: Weight of wood

8.81

8.64 8.97 9.17 8.97

SI weight, g / 100g chest weight 40.19 39.70 40.97 43.96 38.69

SI length, cm / 100g chest weight 84.86 "81.97" 85.65 "93.70 ** 84.86 'ab P <.05

Inv = Invention

SI = Small Intestine (small intestine)

TABLE 10 TASTE EVALUATION

Number of correct identifiers j * Tissue Comparison Groups

Experiment 1 Experiment 2 Combined

Leg / thighs

Stafac ”10 in Control, P 6 Stafac” 10 in ΧΙΝΟΟ, Ρ 3

Dirty germs, P and Control, P 2

9

7

B »10

Chest

Stafac ”10 in Control, P 2 Stafac ^R 10 in XINOC, P 1

Touch Microbes, P and Control, P - 5 * The odds were detected by the assessors statistically frequently enough (P <.05) ί The number of correct identifications of the unusual pattern required for significance at 5% was 7 for n = 10 and Π for n = 20

An experiment with broilers was conducted to determine the effectiveness of microbes for direct use in diets in mushy and pelleted foods. Regardless of the process, microbes for direct nutritional use improved (P <.05) the conversion of food through Control itself, while the weight increased through Control only in mushy foods. Microbes for direct nutritional use, P improved (P> .05) conversion via Stafac * 10, which was similar to Control, P. Microbes for direct nutritional use, M improved (Ρ? ^ · .05) breast weight and thoracic yield via Control, M while microbes for direct nutritional use, P showed improvement (P> .05) via Control, P. With microbes for direct nutritional use, P treated birds did not, in compared to Stafac ^R 10 treated birds produced no taste changes.

Example 6

Forty-four brown fattening pigs (an average weight of 18.82 kg) were randomly selected and distributed in brick floor enclosures by weight and sex (Table 11) and fattened for 119 days. The composition of the rations of the basic food in the growth phase and in the final phase is shown in Table 12. The rations in the growth phase were given until the average weight of each enclosure was 54.43 kg and then fed with the final phase food until slaughter. All diets contained Mecadox ^R (50 g / t) up to a live weight of 34.02 kg. Control and selected microsphere microbe cultures for direct use in the diet were used at 1 x 13 * CFU / g of food. All meals were given in the form of porridge. Six similar enclosures of twelve fattening pigs were used with each experimental meal.

At the start of the study, we administered the pigs to Ivomec ^R for control of internal and external parasites. Sfaguard ”was added after four weeks to control the worms.

Body weight, food consumption and mortality were recorded on the fences. For each fence, however, we also calculated the hr ane conversion.

Prior to this study, the whey cultures of the microspheres were expanded with calcium carbonate. The theoretical number was 2 x 13 ⁷ CFU / g of product. An 11 g sample was analyzed in duplicate to determine the actual number of product. The sample was paid using a standard plating technique for microsphered lactic bacteria.

An eiodate sample of 1 g of product was analyzed in duplicate to verify the product number and composition of the genera.

Samples were taken weekly for each treatment and tested for microsphered lactic bacteria.

The product was confirmed as well as the desired levels of organisms in it (Table 14).

Sample resuscitation varied from 1 x 10 * to 1.6 x 10® CFU / g feed (Table 15). Two extreme samples were attributed to the sampling error. "Payment. Other samples averaged around the target level of 1 x 10 * CFU / g feed.

Microsphere products improved (P> .05) weight and food conversion via Control after 28 days (Table 13). The pigs were hit by the TSE outbreak the first week of the experiment. This outbreak is likely, together with the time required for the pig's digestive tract to adapt to the product, the reason for the 28-day lag behind detectable progress. It is apparent that microbes of the microspheres for direct use in the diet of this invention have an effective effect on pigs as well as on chickens and turkeys,

The above examples show that the invention achieves all of the stated goals,

TABLE 11

Fence Arrangement Attempt: 670-9102

Processing Fence numbers

Control 3, 4, 6, 9, 11, 12

Dirty germs. up 1, 2, 5, 7, S, 10 * Fence size 1.4 m m 4, B8 m, one Smidley four-hole controller, one nipple feeder, heat control spreaders, partially brick floors and a modified front open building.

TABLE 12

Composition of Basic Meals Experiment: 670-9102

Ingridients Production stages Growth Conclusion Basic cereals 76.30 82.20 Soybean meal 21,25 15.50 Bicalcium phosphate 1,05 0.90 Calcium carbonate 0.85 0.85 Sol 0.30 0.30 Vitamin / mineral premix 0.25 0.25

Table 13

Fence Production Data Experiment s670-9102

Control

Microbes to use

Day 14

Weight yield, kg 4,35 4,17 Food conversion 2,439 th most common 2,483 th most common Mortality 1.41 1.39 Day 28 Weight yield, kg 11,93 12,65 Food conversion 2,405 th most common 2,212 th most common Mortality, 7. 2.82 1.39 Day 42 Weight yield, kg 20,77 21,23 Food conversion 2,497 th most common 2,428 th most common Mortality, 7 2.82 2.78 Day 56 Weight yield, kg 31,34 32.43 Food conversion 2,507 th most common 2,457 th most common Mortality, 7 4.23 2.78 Day 70 Weight yield, kg 40.55 41.37 Food conversion 2,735 th most common 2,674 th most common Mortality, 7 5.63 2.78 Day 84 Weight yield, kg 50.35 50,80 Food conversion 2,904 th most common 2,882 th most common Mortality, 7 5.63 2.78

Day 98

Weight gain, kg Food conversion Mortality, X

Day 112

Weight yield, kg Food conversion Mortality, 7.

Day 119

Weight yield, kg Food conversion Mortality

58.92

3,071 th most common

5.63

69.04

3,164 th most common

5.63

73.66

3,217 th most common

5.63

60.78

2,988 th most common

2.78

70.17

3,134 th most common

2.78

75.11

3,177 th most common

4,17

Table 14

Production CO and CA Trial: 670-9102

Product QC number OA number Ratio of genera CFU / g of product SF202: SF301 Dirty germs. use in prehr 4.3 χ 10 ^

Table 15 QA Fences Experiment: 670-9102

Date Control Dirty germs. use 5/3/91 1.7 x 10 ³ 7.4 X 10 ³ 5 / 8/91 1.8 x 10 » 1.6 x 10 » 5/22/91 0 1.0 x 10 ^x 5/30/91 0 2.6 χ 10 ³ 6 / 5/91 2,0 x 10 = 9 · * - χ iS ⁴ 6/12/91 1.0 x 10 * 2.0 x 104 6/19/91 0 9.6 χ 10 ³ 6/26/91 6,7 x 10 = 5,6 χ 10 ³ 7 / 3/91 4.9 x 10 = ~ T o χ 10 ³ 7/10/91 5,2 x 10 ^x 3.0 4 10 * 7/17/91 1.0 x 10 = 4.5 χ 10 ³ 7/24/91 0 1.0 X 10 * 7/31/91 0 1,2 x 10 * 8/7/91 0 6.4 x 10 * 8/14/91 0 7.7 χ 10 ³ 8/21/91 0 1,2 x 10 * 8/28/91 . 0 9.1 χ 10 ³ 9/4/91 0 2.9 x 10 * 9/18/91 0 8.9 χ 10 ³ 9/25/91 0 5.5 χ 10 ³ An asset 9,5 x 10® 8.4 χ 10 ³

FOR

PIONEER HI BREAD, 700 Capitol Square,

INTERNATIONAL INC. 400 Locust Street

Claims (18)

1. A method of promoting animal growth, characterized in that it consists of; supplementing normal animal rations with small but accelerating growth of effective amounts of microbes for direct nutritional use consisting essentially of the life-sustaining, stable, dried microspheres of the fatty acids Enterococcus faecium 301, ATCC St._, and for the life of enterococcus faecium 202 capable, stable, dried microspheres of fatty acids, ATCC St .---- 2. The method of claim 1, wherein the fatty acid spheres are formed using a rotating disk. The method of claim 2, characterized in that the microbes for direct use in the diet of approx. 30% to 70% of one of the mentioned fatty acid microspheres, the rest being the other. 4. The method of claim 3, wherein the fatty acid is C _i2 DD C ₂ ^. free fatty acid » 5. The method of claim 4, wherein the fatty acid is stearic acid » 6 "The method of claim i, characterized in that all said streptococci are present in equal amounts. The method of claim 1, characterized in that the amount of microbes for direct use in the diet, which is added to food rations from about 0.2267 kg to approx. 0.9072 kg / tonne of food. 8 ,. The method of claim 7, characterized in that the amount of microbes for direct use in the diet is from about 0.3629 to approx. 0,5443 kg / tonne of food. 9. The method of claim 7, wherein the number of microbial organisms for direct use in the diet is from approx. ί x 10 »CFU / gm to approx, 2 χ 10® CFLi / gm. The method of claim 9, wherein the number of microbial organisms for direct use in the diet is approx. 1 x 10 »CFU / gm 11. The method of claim 1, wherein the suture is chicken. 12. The method of claim 1, wherein the animal is a pig. Composition of microbes for direct use in the diet to promote the growth of pig animals, characterized in that it consists essentially of the viable, stable, dried microspheres of the fatty acids Enterococcus faecium 301, ATCC and the viable, stable, dried microspheres of the fatty acids Enterococcus fa ec mm 202. 14. Microbes for direct nutritional use according to claim 13, characterized in that they contain from approx. 30% to approx. 207. one of the streptococci mentioned, the remainder being the other. 15. Microbes for direct nutritional use according to claim 14, characterized in that the free fatty acid C _X3 DO is free fatty acid. 16. Microbes for direct use in the diet of claim 15, Α · <4 characterized in that free fatty acid is stearic acid. Microbes for direct use in the diet of claim 16, characterized in that the streptococci organisms are present in equal doses. The composition of claim 13, wherein the animal is a chicken. The composition of claim 13, wherein the animal is a pig. FOR PIONEER HI BREAD, INTERNATIONAL INC. 700 Capitol Square, 400 Locust Street Des Moines, Iowa 50309 USA. Jgpko Ručntik ABSTRACT Enterococcus F.aecimiG genera 301 and 202 are dried and used in animal feed additives to promote the growth of animals and improve the quality of animal skeletons using the rotating disk of the dried microspheres of the Enterococcus F.aecimiG fatty acids 301 and 202. FIGURE 1 YOU ARE THE NUMBER OF MEASUREMENTS Trn ττιτρ ττππτΓ | ΓΓτπππρτΓΓΓΓτΐ ϊ j i 11 j i n π, 1111 f ιπι j 11 r n n ii | 10 20 30 DAYS OF MEAT