WO1994011492A1 - Procede modifiant de maniere avantageuse la microflore intestinale de la volaille - Google Patents
Procede modifiant de maniere avantageuse la microflore intestinale de la volaille Download PDFInfo
- Publication number
- WO1994011492A1 WO1994011492A1 PCT/US1993/010644 US9310644W WO9411492A1 WO 1994011492 A1 WO1994011492 A1 WO 1994011492A1 US 9310644 W US9310644 W US 9310644W WO 9411492 A1 WO9411492 A1 WO 9411492A1
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- Prior art keywords
- feed
- poultry
- spores
- fed
- intestinal
- Prior art date
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Classifications
-
- 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
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- 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
- 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
- C12R2001/07—Bacillus
Definitions
- the general field of this invention is direct-fed microbials for animal feeding.
- the present invention is more particularly concerned with direct-fed microbials for poultry.
- Direct-fed microbials are live microbial cultures supplemented into feeds which contribute well-being to animals by improving their intestinal microbial balance. All direct-fed microbials have as their active ingredient live bacterial cells. Species of the genera Lactobacillus, Streptococcus, and Bifidobacterium have been employed as direct-fed microbials. The major mode of action for these bacteria is production of organic acids especially lactic acid which can inhibit growth of pathogenic or potentially pathogenic bacteria.
- Modifying the intestinal microflora of poultry such as chickens, turkeys and ducks, presents special problems that are different than those encountered with domestic mammals, such as cattle, sheep and swine.
- Coliform organisms are ubiquitous in the intestines of poultry, and if their growth is not controlled, the coliforms can multiply and have adverse effects on the health and growth of poultry.
- Poultry are also especially subject to infection with Coccidia which can produce intestinal lesions which predisposes the bird to coliform proliferation. For this reason, coccidiostats are commonly included in poultry feeds. It is therefore important for a poultry bacterial feed additive to be compatible with coccidiostats.
- Bacterial additives in poultry feeds have not been extensively studied. However, it has been reported that some benefit can be obtained by including Bacillus subtilis cells in turkey feeds: Jaraphocakul, et al. (1990), Poultry Sci.. 69:1966-1973.
- Bacillus subtilis cells in turkey feeds Jaraphocakul, et al. (1990), Poultry Sci.. 69:1966-1973.
- body weight and feed efficiency of the turkeys were not significantly affected.
- Microbiological testing disclosed that the B ⁇ _ subtilis counts in the crop and cecum were increased by the feeding, but the B_j_ subtilis feeding failed to influence intestinal Lactobacillus or Escherichia coli.
- limited increases in body weight and feed efficiency were observed. Turkeys receiving the IL_ subtilis showed an increase in body weight gain at 12 weeks, and the feed efficiency showed improvement at 20 weeks.
- Chr. Hansen's Laboratory, Inc. of Milwaukee, Wisconsin, USA has marketed a direct-fed microbial additive under the trademark "BIOMATE 2B", which contains a mixture of Bacillus subtilis and Bacillus licheniformis. That additive has been used with domestic animals including poultry.
- Chr. Hansen's Laboratory has carried out research investigations to find a lactic acid-producing bacteria which is especially beneficial when fed in a mash or pelletized feed to poultry. The present invention is a result of that research investigation.
- U.S. Patent 4,919,936 of 1990 discloses a strain of Bacillus subtilis (C-3102; FERM BP-1096) , which is said to provide improved results with a wide variety of domestic animals including poultry.
- Example 3 (cols. 4-5) of this patent reports experiments with broilers and laying chickens. An increase in body weight and feed conversion was obtained for the broilers, and the number of eggs and egg weights were increased for the laying chickens. No examinations of intestinal microflora were reported.
- U.S. Patent 4,999,193 of 1991 discloses a strain of Bacillus cerus (IP5832) for improving the growth of domestic animals.
- B. cerus forms vegetative spores which can be incorporated in pelletized feeds, the spores being sufficiently heat resistent to survive a pelletizing process.
- the claimed beneficial effects are improvements in feed conversion and growth.
- Example 3 (cols. 3-4) of this patent describes an experiment with female chicks where the live weight and consumption index was improved by the feeding of the B. cerus strain. No data relating to intestinal microflora was presented.
- Bacillus pumilus is a spore-forming lactic acid-producing bacteria, which produces acetylmethyl- carbinol, coagulates milk, and ferments gelatin, and does not grow well at 60°C.
- Bacillus coa ⁇ ulans is also a spore-forming, lactic acid producing bacteria, which also produces acetylmethylcarbinol, coagulates milk, grows variably at 60°C, but does not ferment gelatin.
- the properties of B ⁇ _ pumilus or B_ 5 _ coagulans as direct-fed microbial feed additives have not been previously reported. As far as it is known neither B. pumilus nor EL. coagulans have been used in a direct-fed microbial feed prior to the research studies of Chr. Hansen's Laboratory.
- Bacillus pumilus has properties which make it especially desirable for incorporation in a direct-fed microbial poultry feed.
- B. pumilus has the ability to produce lactic acid after being pelleted in a feed and has other important properties.
- B ⁇ . pumilus can withstand acid conditions, and can germinate quickly from spores in the intestines of poultry. This contrasts with bacteria from the genera Lactobacillus, Streptococcus, and Bifidobacteria which do not appreciably produce lactic acid after being pelleted in a feed.
- B_ s _ pumilus spores By incorporating B_ s _ pumilus spores in poultry feeds, the intestinal microflora can be favorably regulated. Among other observed results, intestinal coliform organisms can be appreciably reduced. It has also been found that B pumilus and/or B_j_ coagulans can be incorporated in poultry feeds containing coccidiostats for conjoint action. Not only does B. pumilus and coccidiostats not interfere with each other's actions, but they appear to somewhat act synergistically in preventing formation of lesions due to coccidia, and in improving the health of the birds, resulting in reduction in mortality. Feed efficiency and growth rate are also significantly improved. Further, due to the enzymatic action of B ⁇ _ pumilus on the ingested feed, litter quality is improved as evidenced by less wet, caked litter, better litter score, and lighter litter weight.
- the method of the present invention in which B ⁇ . pumilus spores are incorporated in poultry feeds can improve health, reduce mortality, improve litter quality, and at the same time improved feed utilization.
- a further advantage is that these spores can be conveniently administered.
- the spores can be premixed with a phosphate salt which is to be incorporated in the feed, the phosphate salt acting as a diluent and carrier for the spores. It is not necessary to contol the water activity of the phosphate salt because the bacteria are in spore form.
- the spores, with or without premixing with a carrier can be incorporated in pelletized feeds.
- Bacillus pumilus is the preferred bacteria for use in practicing the method of this invention. Alternatively or additionally, Bacillus coagulans can be used. Both species are spore-forming and produce lactic acid.
- the direct-fed microbial properties of B. pumilus has not been considered prior to this invention, although it is a Bacillus species that can be ingested by domestic animals.
- Usable strains of B. pumilus or B coagulans are available from public depositories, such as the Northern Regional Research Laboratory, U.S.D.A., Peoria, Illinois, USA. Available strains from this source include Bacillus pumilus NRRL NRS-272 and Bacillus coagulans NRRL-B768.
- Bacillus pumilus or B_j_ coagulans can be obtained and used.
- the BC 235 strain has been found to have the characteristics of the species of B. pumilus; viz. it does not grow at 60°C (growth stopping at 55 ⁇ C) , it does not grow well anaerobically, and it ferments gelatin, and has other characteristics by typing.
- the poultry feed is modified by incorporating an effective concentration of viable spores of Bacillus pumilus and/or coagulans.
- the modified feed is then fed to poultry in an amount and for a length of time sufficient to appreciably modify their intestinal microflora.
- This modification of intestinal microflora can be verified by microbial examination of the intestinal contents of killed birds, for example, by determining that a reduction in coliform organisms has occurred.
- the amounts to be fed and the feeding periods can predetermined and standardized.
- the modified feeds used for the purpose of the present invention can contain from 10 3 to 10 8 viable spores per gram of feed. In presently preferred embodiments, the feeds contain from 10 5 to 10 7 viable spores per gram of feed. For example, a preferred level for commercial use is approximately one million spores per gram of feed.
- the method can be practiced with a wide variety of poultry being raised for commercial purposes. Important species include chickens, turkeys, and ducks. The method can be used with particular advantage with poultry being raised for meat production, but its beneficial effects are not limited to this class of birds. The method is generally applicable to male, female and caponized birds, and can be used effectively with breeder birds and egg laying birds. Further, the benefits of the method are not limited to a particular growth phase, but can be used advantageously with starter, grower and finisher birds.
- the modified feeds are administered on a daily basis, for example, continuously for several weeks.
- feeding is continued for at least three weeks, such as from one day of age to three weeks of age.
- the length of feeding will depend on the kind of poultry and the purpose for which they are being fed.
- Propagation of Bacillus pumilus or coagulans can be carried out according to known methods for growing Bacillus species.
- stock cultures may be propagated in a trypticase soy broth fortified with yeast extract and minerals.
- the propagation medium can be adjusted initially to a neutral pH. Fermentation is carried out with aeration and agitation, at a temperature of around 37°C, and pH control is preferably used.
- Fermentation is carried out with aeration and agitation, at a temperature of around 37°C, and pH control is preferably used.
- the cells go through a logarithmic growth phase. After the nutrients begin to become depleted, the cells convert to spore form. Then the spores can be harvested by centrifugation and lyophilized for incorporation in poultry feeds.
- the spores can be admixed with a suitable carrier for addition to the feed ingredients.
- a phosphate salt can be used as a carrier, since phosphate salts are necessary ingredients of poultry feeds.
- the phosphate salts do not need to be prepared in any special way.
- spores can be mixed with commercial phosphates such as dicalcium phosphate, defluorinated phosphate, etc.
- the desirable proportions of spores to phosphate salts range from about 10 to 10 spores per gram of phosphate salt.
- Poultry feeds are usually designed as complete rations. These feeds comprise mixtures of solid feed ingredients in the form of powders or granules. When the feed mixture is to be pelletized, a liquid ingredient may be included such as molasses. The spores should be blended substantially uniformly with the feed ingredients. Thereafter, if desired, the feed can be pelletized, or it can be fed as a loose or mash-type feed.
- Propagation of Bacillus pumilus or coagulans can be carried out to obtain the bacteria in spore form.
- stock cultures are propagated overnight at 37°C in trypticase soy broth before being inoculated into production medium.
- Production medium can consist of meat peptone (2%) , yeast extract (0.5%), K 2 HP0 4 (0.1%), MgCl 2 * 6 H 2 0) (0.02%), MnCl 2 ' 4 H 2 O (0.01%), CaCl 2 (0.02%), and defoamer (0.1%); pH was adjusted to 7.0 using 50% NaOH.
- Parameters of the fermentation are 37°C, 0.5 wm aeration, agitation to maintain greater than 25% dissolved oxygen, and pH was controlled with 25% H 2 S0 4 or 50% NaOH.
- a dextrose (50%) feed is used during fermentation with a delivery rate of 3.8 gram per hour for the first 16 hours of fermentation.
- the fermentation is terminated when the culture is considered to be completely sporulated as determined by vesical microscopic examination.
- the spores thus produced are separated from the fermentate by high speed centrifugation and lyophilized.
- Test Methods Day old turkey poults were weighed, wing banded and then placed in their appropriate pens. Treatments included: 1) a typical starter turkey ration (control diet) and 2) this same diet but containing viable BC-235 as a feed additive. The spores were added to the control diet by using dicalcium phosphorus as a carrier, the spores being premixed with the carrier. Approximately 1 trillion spores were added per ton (2000 lbs.) of feed. Birds were observed daily for signs of morbidity and(or) mortality and weighed at 7, 14, and 21 days of age. At the conclusion of the trial, representative intestinal samples were enumerated for intestinal bacteria.
- Test Results Mortality was reduced in birds fed the BC 235 spores when compared to the control birds (0.0% vs. 4.2%). Total body weight gain for the 21 day test period was greater for the birds fed the feed additive (23.32 lb. vs. 22.79). Feed efficiency for the 21 day period was improved in birds fed the feed additive (1.781 vs. 1.538). The spores decreased the concentration of intestinal coliforms from 12 million coliforms per gram of wet intestinal contents to 3 million coliforms per gram of wet intestinal contents.
- the intestinal spore count an indicator of the presence of BC 235 spores, was increased in birds fed the feed additive from 7100 spores per gram of wet intestinal contents in the control birds to 190,000 spores per gram of wet intestinal content in the treated birds.
- the spores were added to the diet via a non-conditioned, defluorinated phosphate carrier, being premixed therewith, to deliver approximately 1 trillion spores per ton (2000 lbs.) of feed.
- Birds were weighed at 21, 42 and 47 days of age. At day 10 of the study, birds were evaluated for the incidence of pasted vents. At day 14, all birds were orally challenged with coccidia and then on day 21, representative birds were evaluated for intestinal lesions. On days 14, 21 and 47, representative birds were evaluated for intestinal spore counts (detection of the feed additive) and coliforms. At the conclusion of trial, litter quality was evaluated.
- Test Results Data are pooled across coccidiostats to compare the overall effect of feeding the feed additive to broilers.
- the BC 235 additive improved feed conversion compared to the diets not containing the feed additive (1.829 vs. 1.841) and tended to improve the overall feed conversion at day 47 (1.911 vs. 1.920) .
- the feed additive decreased mortality from 3.05% to 2.01% and reduced the incidence of intestinal lesions caused by the coccidia challenge from 1.96 to 1.665 (an average score across all intestinal sections) .
- the feed additive also improved litter quality as evidenced by less percentage of wet, caked litter (47.9% vs. 48.7%), better litter score (1.6 vs.
- the intestinal spore data on day 47 indicated that the BC-235 spores were viable in the intestinal tract (11,000 spores per gram of wet intestinal conents vs. 6,000 spores per gram of wet intestinal contents) , and the coliform population reduced by 25% (9.5 million per gram of wet intestinal contents vs. 12.5 million per gram of wet intestinal contents) .
- the pelleting data indicated that BC 235 spores survived pelleting at 180°F.
- BC 235 spores were viable in the intestinal tract in the presence of coccidiostats, reduced intestinal coliform counts, when administered in feeds pelleted at 180°F.
- B. pumilus or coagulans spores can be fed to poultry in conjunction with other helpful bacteria.
- Bacillus licheniformis and Bacillus subtilis strains can be used to produce significant amounts of amylase and protease enzymes in the poultry intestines.
- a combined direct-fed microbial additive can be prepared as follows: Composition of the feed additive per 1000 g: Bacillus subtilis spores 50 g
- Bacillus pumilus or coagulans spores 100 g
- the feed additive of the foregoing composition was used in a comparative study of chicks of older hens and chicks of younger hens.
- Feeds were formulated and mixed in batches sufficient for one week's worth of feeding. Diets were formulated to meet National Research Council recommendations. All starter and grower feeds contained Amprol Plus at a rate of 0.0125% amprolium and 0.0004% ethopabate. Withdrawal feed, which was fed for the final three days of the trial, contained no medication. The feed additive as described above with BC 235 spores was mixed with the feed prior to pelleting (180°F) at a rate of 2 x 10 11 viable bacteria per ton of feed. Broiler starter feed was fed from day 0 to 21, grower/finisher feed was fed from day 21 to day 39, and withdrawal feed from day 39 to 42.
- Test Results Chicks from old hens responded to the feed additive with lower mortality rates than control chicks. Treated chicks from younger hens did not show lower mortalities until week 6. Relatively little effect was observed on final body weights, feed intake, or feed efficiency. This test showed a benefit of the present invention in lowering mortality in broiler chicks.
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Abstract
On fait ingérer à la volaille le Bacillus pumilus, le Bacillus coagulans, ou des mélanges de ceux-ci sous forme de spores afin de modifier sa microflore intestinale. L'on réduit ainsi des organismes coliformes potentiellement dangereux et l'on obtient d'autres résultats avantageux tels que l'amélioration de la santé de la volaille, une mortalité réduite, l'amélioration de la qualité de la litière ainsi qu'une meilleure alimentation.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US97561092A | 1992-11-12 | 1992-11-12 | |
US07/975,610 | 1992-11-12 |
Publications (1)
Publication Number | Publication Date |
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WO1994011492A1 true WO1994011492A1 (fr) | 1994-05-26 |
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ID=25523194
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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PCT/US1993/010644 WO1994011492A1 (fr) | 1992-11-12 | 1993-11-02 | Procede modifiant de maniere avantageuse la microflore intestinale de la volaille |
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