Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
  • A61K35/66—Microorganisms or materials therefrom
  • A61K35/74—Bacteria
  • A61K35/741—Probiotics
  • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K10/00—Animal feeding-stuffs
  • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
  • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K40/00—Shaping or working-up of animal feeding-stuffs
  • A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K50/00—Feeding-stuffs specially adapted for particular animals
  • A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/20—Bacteria; Culture media therefor
  • C12N1/205—Bacterial isolates
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
  • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
  • C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
  • C12R2001/00—Microorganisms ; Processes using microorganisms
  • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
  • C12R2001/46—Streptococcus ; Enterococcus; Lactococcus

Definitions

• Growth enhancers in the form of antibiotics have been used extensively for poultry, namely chickens and turkey.
• Growth enhancers such as Stafac ® and BMD ® (bacitracin methylene disalicylate) are known.
• antibiotics and have been used at sub-therapeutic levels of for example, 10 grams per ton and 25 grams per ton as feed additives in order to promote
• Direct-fed microbials do have some difficulty in maintaining a stable product.
• the direct-fed microbial is used at a fairly low level, added to feed at perhaps a 0.1% level.
• unused direct-fed microbial containing feed or feed additive product is often stored by the farmers for long periods of time. This storage many times is under conditions where there is some moisture and high temperature. In many instances there is just enough moisture that the bacteria are activated or start to grow, but yet there is an insufficient amount of moisture to sustain them. As a result they die. Thus, the activity of the direct-fed microbial is stopped. In other instances, the
• Uniformity of flock weight is important because if more birds are normal in size, less hand labor is required and processors can more extensively rely on machine processing. On the other hand, if the birds vary considerably from very small birds to very large birds, even though the overall flock weight may be the same, the smaller birds and the larger birds require a great deal more hand labor and because of their lack of uniformity in size, cannot be processed easily by machine. Thus, uniformity of flock weight with a high percentage distribution within the normal size range so that chickens can process by standardized machinery is a desirable feature.
• invention to provide a direct-fed microbial which, for poultry, provides increased rate of weight gain, which provides better feed conversion, which provides higher yield of breast meat, and which provides for
• An even further primary objective of the present invention is to provide direct-fed microbials suitable for poultry feed ration addition which contains bacteria that are in microsphere form using a special rotary technique using free fatty acid matrix.
• Another objective of the present invention is to provide a direct-fed microbial which has stability at levels within the range of from 3 months to 6 months without any significant organism count reduction.
• Another objective of the present invention is to provide a process of rotary formation of spheres containing the dried bacteria which provides having uniform size.
• Another objective of the present invention is to provide rotary disc spheres of dried bacteria which are free flowing, and easily processable with poultry feed rations.
• a still further objective is to provide a
• microsphere of fatty acid material containing certain bacteria with spheres being useful as a direct-fed microbial for both poultry and swine.
• Figures 1, 2 and 3 show graphically the stability of the strains using stearic acid matrix.
• Figure 4 is a graph showing breast yield
• Figure 5 is a graph showing body weight
• Figures 4 and 5 show a control, use of an
• the invention is a method and composition of growth promotion for poultry and swine which comprises adding to the normal feed ration a small but growth promoting effective amount of a direct-fed microbial which contains dried, fatty acid microspheres of
• Enterococcus faecium 301 DSM No. DSM-Nr. 4789, and dried fatty acid microspheres of Enterococcus faecium 202, DSM No. DSM-Nr. 4788, where preferably the fatty microspheres are formed by rotary disc drying.
• a fatty acid employed may be any one of the C 12 to C 24 free fatty acids, but is preferably stearic acid.
• the organisms are preferably present in about equal amounts but may vary within the range from about 30% to about 70% of one of the organisms with the balance being the other.
• the amount of direct-fed microbial added to the feed ration can vary considerably but generally will be within the range of from about 0.5 pounds to about 2.0 pounds per ton of feed, generally from about 0.8 pounds to about 1.2 pounds per ton of feed, and typically at about 1 pound per ton of feed.
• the organism count, that is the number of colony forming units per gram present in the direct-fed microbial can also vary within the range of from about 1 ⁇ 10 6
• CFU/gm to about 2 ⁇ 10 9 CFU/gm, but is preferably at about 2 ⁇ 10 8 CFU/gm.
• Stafac ⁇ and BMD The advantages of sub-therapeutic levels of antibiotics as growth promoting additives can be achieved with naturally occurring organisms of the present invention provided that direct-fed
• microbial is made in accordance with the present invention and added in accordance with the method described herein.
• a combination of direct-fed microbial and growth promotant together exceeds the advantages of either alone and thus they may be used together if desired.
• the method of processing of the organisms is not critical as long as the organisms can be kept alive to delivery to the animal, and placed in a form so that it will combine with animal feed well and is of a generally uniform size so that dosage may be
• a microsphere refers to a fatty acid matrix in which a plurality of organisms is incorporated. It is different from a microcapsule in which individual organisms are each encapsulated. In a microsphere the fatty acid matrix functions for the composite similar to the
• microspheres are formed wherein each sphere constitutes a plurality of bacteria in a free fatty acid matrix rather than an individual
• microencapsulator of each bacteria in a coating or film like layer of fatty acid This provides
• the preferred matrix agent is a C 12 to C 24 free fatty acid. While mixtures of fatty acids may be employed, it is preferred that a single pure free fatty acid be employed. It is also preferred that the free fatty acid be an unsaturated fatty acid, with the most preferred being stearic acid.
• the fatty acid have a melting point less than 75oC, preferably within the range of 40°C to 75°C. It must, of course, be solid at room temperature in order to be an effective matrix. All free fatty acids falling within the range of chemical description heretofore given will meet these requirements.
• the bacteria are typically freeze-dried bacteria as placed in the product. Thus, they can be revived by moisture addition.
• the microspheres generally comprise from about 50% to over 90% by weight of the fatty acid component with the balance being bacterial culture.
• the preferred range is from about 60% to about 75% fatty acid. If too little fatty acid is used, the matrix will be inadequate for protection. On the other hand, if too much is used, the matrix will be too thick and results in inadequate release in the gut.
• the process as used in this invention is a rotary disc microsphere formation process.
• a slurry of the bacteria and fatty acid components are thoroughly mixed with the mixture being added at a uniform rate onto the center of a rotating stainless steel disc. It is there flung outwardly as a result of centrifugal force and forms a microsphere. It is then collected in a cooling chamber maintained at ambient conditions or slightly lower, sized and readied for packaging.
• microencapsulation In conventional tower spray drying there is a tendency for particles to cluster, for the coating to be uneven, and thus for the
• the rotary disk typically employing a 4"-6" rotary disc
• the rotary disk can be run at the rate of from 2000 rpm to 4000 rpm, preferably about 2500 rpm to 3200 rpm with a feed rate of from 50 grams to 200 grams per minute.
• the preferred conditions presently known are use of stearic acid, use of two hereinbefore described organisms, a four inch rotary disc, 3000 rpm and a feed rate of 100 grams per minute with a bacteria/stearic acid slurry of 35% bacteria, 65% stearic acid.
• a product having a particle size of from 75 microns to 300 microns will be achieved, with a preferred level of less than 250 microns.
• Example 1 through 4 and Figures 1, 2, and 3 relate to the invention of my prior case.
• Example 5, and tables 2-10 relate to the process of this present invention for a poultry direct-fed microbial.
• Example 6 relates to turkeys specifically and example 7 relates to swine.
• Example 1 correlates with Figure 1. It shows the product stability of two different strains of
• Enterococcus faecium with temperatures of 4°C and 27 oC.
• Figure 1 shows a stability of the encapsulated strains of Enterococcus faecium, with the encapsulation being by the rotary disc device using stearic acid with a level of 35% culture weight.
• Conditions of microsphere formation were as previously described herein, namely a 35/65 bacteria stearic acid slurry at a temperature of 60 °C, using a four inch rotary disc, operating at 3000 rpm and a feed rate of 100 grams per minute.
• the spheres were formed, placed in heat sealed vapor barrier pouches and destructively sampled weekly for CFU determination. It can be seen that the product of the invention maintained excellent organism colony forming unit (CFU) counts out to storage times at long as 70 days.
• CFU colony forming unit
• Example 2 is to be interpreted in connection with Figure 2.
• the figure shows the stability of
• Example 3 is to be interpreted in conjunction with Figure 3. It shows the stability of the
• Table 1 is a list of the minimum times for a 1 log loss in colony forming units (CFU). Table 1
• Example 4 the stability of product after pelletizing for use of a chicken feed product was determined.
• the microsphere formation conditions were as earlier described.
• the conditions used in this study were the following:
• Conditioning temperature was 70"C and the pellets out of the dye were 78 °C.
• Pellets were stored in unsealed bags and sampled weekly for CFU determination.
• the pelletized product was not adversely affected in stability by the conditions of pelletizing.
• the pelletized product showed stability equal to the unpelletized product.
• Starter, grower, and withdrawal rations were formulated to contain 1425, 1450, and 1475 kcal ME/lb, respectively, with 90 g/ton monesin. Starter rations were fed from 1 to 21 days of age, grower from 21 to 42 days of age, and withdrawal from 42 to 49 days of age. The
• each rotary disc fatty acid encapsulated as described in Example 1 and ed n. present as 50% of the direct-fed microbial applied at 1 ⁇ 10 5 CFU/g of feed, mash (direct-fed microbial, M); negative control, pelleted (Control, P); direct-fed microbial applied at 1 ⁇ 10 6 CFU/g mash, pelleted (direct-fed microbial, P); and a positive control applied at 10 g/ton
• a mixer test was conducted for each production phase. The test was designed to ensure that the direct-fed microbial was uniformly distributed at appropriate levels in the feed and that it survived pelleting. Each batch was sampled at the time of bagging with 4 equally spaced samples for the mash treatments and 10 equally spaced samples for the pelleted treatments (i.e. bags 1, 3, 5,..., 35, 37, and 39).
• Alternate floor pens within a treatment had non-contaminated feed sampled during weeks 1 and 4; with the remaining pens sampled on weeks 2 and 6 during the feeding study.
• Direct-fed microbial regardless of processing, improved ( P ⁇ .05 ) feed conversion over the respective Control while increasing (P ⁇ .05) weight gain over the Control only in the mash feed (Table 4).
• the product was at its desired level and strain composition (Table 5).
• Direct-fed microbial was uniformly distributed within the feed. Direct-fed microbial, M was at its desired level while direct-fed microbial, P was 1 to 1-1/2 log higher than desired for the starter and grower rations (Table 6). The high counts for direct-fed microbial, P were a result of overengineering of the product to ensure sufficient recovery of the organisms after pelleting.
• the direct-fed microbial treatments produced a shorter (P>.05) small intestinal length than either of the Controls and Stafac ® when expressed as actual length, a ratio of either body weight, or breast weight (Table 9).
• Direct-fed microbial, M had a lighter (P>.05) small intestinal weight than Control, M when expressed as either actual weight or percentage of either body or breast weights.
• the reduction in intestinal weight and length for direct-fed microbial treatments suggests less energy required for
• Mecadox ® 50 g/t up through 75 lb bodyweight followed by 100 g/t
• the commingled feeder pigs were given Ivomec ® to control internal and external parasites.
• Safeguard ® was given after four weeks to control whipworms. Body weights, feed consumption, and mortality were recorded by pen. Feed conversion was calculated for each pen.
• microsphere culture concentrate Prior to the study, the microsphere culture concentrate was extended with calcium carbonate. The theoretical count was 2 ⁇ 10 7 cfu/g of product. An 11 g sample of product was assayed in duplicate to determine actual product counts. The sample was plated using standard plating technique for
• microsphered lactic acid bacteria microsphered lactic acid bacteria.
• Pen sample recoveries varied from 1 ⁇ 10 1 to 1.6 ⁇ 10 5 cfu/g of feed (Table 15). The two extreme samples are attributed to sampling/plating error. The remainder of the samples averaged around the target level of 1 ⁇ 10 4 cfu/g of feed.
• Microsphered product improved (P>.05) weight gain and feed conversion over the Control after 28 days
• Pen size 4.6' x 16.0', one four-hole Smidley feeder, one nipple driker, sprinklers were used for heat control, partially-slated floor, and modified, open-front building.

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• 1993
  • 1993-02-03 CA CA002131790A patent/CA2131790A1/en not_active Abandoned
  • 1993-02-03 WO PCT/US1993/000867 patent/WO1993019162A1/en not_active Application Discontinuation
  • 1993-02-03 EP EP93904784A patent/EP0631616A4/en not_active Withdrawn
  • 1993-02-03 CZ CZ942253A patent/CZ225394A3/cs unknown
  • 1993-02-03 RO RO94-01518A patent/RO112896B1/ro unknown
  • 1993-02-03 HU HU9402673A patent/HUT67965A/hu unknown
  • 1993-02-03 BR BR9306121A patent/BR9306121A/pt not_active Application Discontinuation
  • 1993-02-03 JP JP5516534A patent/JP2849877B2/ja not_active Expired - Lifetime
  • 1993-02-03 SK SK1116-94A patent/SK111694A3/sk unknown
  • 1993-02-03 RU RU94043791A patent/RU2109052C1/ru active
  • 1993-02-24 MX MX9301017A patent/MX9301017A/es unknown
  • 1993-03-17 SI SI19939300128A patent/SI9300128A/sl unknown
• 1994
  • 1994-10-17 BG BG99113A patent/BG99113A/bg unknown

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