CS14192A3 - Method of increasing microbial population in second stomach of ruminantsand of enhancing yield of the animals - Google Patents

Abstract

The process consists in feeding these animals with revivifiable spores of Bacillus cereus, strain C.I.P.5832.

Description

The way of enlarging my animals and improving animals

Technical field

The invention relates to a method for increasing the microbial population

improves the zootechnical performance of these animals. The invention also relates to a feed additive for carrying out this process and a supplementary feed for ruminant animals containing this additive.

BACKGROUND OF THE INVENTION A rumor, which is one of the four stomachs of ruminants, contains a very large number of microorganisms that make up an ecosystem living in symbiosis with an animal host.

These organisms are dominated by the following species: - bacteria, in the order of 10-10 germs per gram of contents, 5 - protozoa, in the order of 10 to 10 germs per gram of tale content.

In addition to these predominant species, they are found in fungi, microscopic fungi, mycoplasma and bacteriophages.

These microorganisms belong to very different genera and species: 22 genera and 63 species can only be described in bacteria (TMYokohama et KA Johnson - DigestivePhysiology and Nutrition - DC Church, publisher, A RestonBook, New Jersey 07532, USA , pp. 125-144). Most of the microorganisms contained in the rum are severe anerobias whose research is difficult.

Due to the difficulties associated with a large number of microbial species present in the tale and due to their aerobic nature, studies of quantitative changes in microbial populations in the tuft are in most cases conducted by microflora compounds.

2 is DAPA (2,6-diaminopimelic acid), which is an amino acid which is specific for microorganisms and which senenaises neither in animal tissues nor in said microorganisms. In a given feeding regime, the proportions of DAPA in the contents of the tonic remain stable and the use of these labeled compounds is particularly useful in measuring the quantitative changes of the microbial population in the tonic. Researches of the microbial population in the tad can be carried out directly on animals in vivo (puncture puncture, fistula- tion and cannulation of the tum, etc.).

Tumor microorganisms are able to degrade and ferment complete feed ruminants that lose an important portion of the nutrients contained therein. Microbial lumps of the tuft microorganisms are also an important feed benefit for ruminants. An increase in microbial populations in the tusk is therefore advantageous for zootechnics of ruminant animals.

Antibiotics and, in particular, ionophore antibiotics are capable of modifying microbial fermentation in the rum. Japanese Patent No. 1977-8381 discloses that Bacilluscereus, a T7H2 strain, is capable of correcting the symptoms of anomaly in fermentation in a ruminant mammal. Such anomalies that cause bloating and putrefaction in the stomach are characterized by an increased amount of ammoniacal nitrogen or propionic acid in the bud content. These features and the results of their treatment are set forth in the following Tables 1 and 2. 7 In the case of bovine animals, overexposure (Table 1) and the amount of ammoniacal nitrogen, which is greatly increased in sick animals, will adjust normal amounts in treated animals. In the case of swelling occurring in lambs, the amount of propionic acid that is increased in the diseased animals is adjusted to normal levels in the treated animals. However, Japanese Invention No. 1977-8381 does not disclose whether Bacillus cereus, strain T7112, has any effect on the shape or composition of a microbial population that is originally in the rum.

Table 1 - cattle

Beef Cattle Ammoniacal Nitrogen f / ml Total Nitrogen mg / ml Volatile NaOHO, 1N lactic acid ^ / ml 1 62 0.74 4.35 51 2 64 0.76 3.25 0 Healthy 3 67 0.84 7, 44 46 4 120 0.71 3.13 56 5 232 0.76 4.50 0 Affected 6,204 0.76 3.50 10 flatulence 7,273 1.36 6.32 50 8,344 0.80 6.20 16 Q 0,5 194 0.53 1.55 8 Flatulence 8 17 0.62 10.62 27 after treatment 9 52 0.62 5.55 74

Table 2 - Lambs

Lambs Acetic acid% Propionic acid% Butyric acid% Other volatile fatty acids% 1 57 Σ3- —19- -5- Healthy 2 59 17 21 3 Diameter 5S 17 20 7 3 56 22 19 3 Impaired 4 49 25 23 3 ouváním 5 56 21 17 6 Average 54 23 20 4. O 58 o 20 4 Treated 4 'C- * 55 16 18 11 5 56 19 21 4 ». ............ Average 56 18 20 6 Μ- In European Patent Application 88108549.2 of 25.05.56 (Priority: 11.06.87 SE 5702435) it is stated that by administering the pacifier to acid producing bacteria Milk (lactobacillus, Pediococcus, Streptococcus and Leuconostoc), or amylase and protease producing species (Bacillus subtilis and Bacillus toyoi), milk composition can be adjusted in dairy cows. a modest increase in the solids content, also in the said specification, is described. However, the results presented in this document (Table 3) clearly demonstrate that only the protein content of milk increases, the milk yield (kg of milk produced per day) does not change and the milk yield corrected for the (standardized) fat content decreases considerably. This means that the fat content is reduced by treatment.

Table 3 h ..... u '- ------------ From Farm 1 Treatment Farm 2 Treatment Farm 3 Treatment 1 1 2 1 2 1 2 í Milk production i ø kg / day 24,71 24,07 19,57 18,91 28,21 28,39 j Milk production I corrected for 1 fat substance kg / day 27,90 22,93 22,74 20,53 29,43 26,65 í * Protein, % 3,47 3,71 3,30 3,63 3,37 3,5 * i Treatment 1: control, no addition of bacterial preparation Treatment 2: with addition of bacterial preparation No reference is made to newalg in European Patent Application No. 88106549.2 into the stomach of the introduced bacteria (increased protein content in the milk) due to any influence of these bacteria (and the original microbial population contained in the barium).

SUMMARY OF THE INVENTION

It has now been found and is the object of the present invention to provide very small amounts of re-invisible spores of Bacillus cereus, the CIF 5232 strain (CI? = Collection de l? Institut Pasteur), into the tincture of ruminants, and increase the content of DAPA in the tincture and increase the zootechnical utility chewing animals (milk or meat production). cloudy amounts of Bacillus cereus spores, C.I.F. 5632, added to the animals, are calculated so that the spores are present in the tuft of the tusk in the amount of 10,000 to 100,000 spores per liter of rum-shave. These amounts are much less than the amounts reported so far for microbial populations. The following 7 examples illustrate the invention without limiting it. EXAMPLES OF THE INVENTION

Experiment on cow with artificial fistula

4 fistled cows are used, a cross-breed Frison-x-Holstein breed, carrying a cannula that allows access to the tory. The animals are fed a daily dose of 9 kg of dry matter, consisting of 30% ensiled maize and 70% concentrated feed. The dose is given to each animal individually at 10 am in the morning. The distributed dose is consumed during the day.

The nutritional characteristics of ensiled maize and concentrated feed are shown in Table 4 below

Table 4

Silage maize Concentrated fodder Dry matter, g / kg 330 S7O Gross cellulose, g / kg 231 109 Amidone + sugars, g / kg 149 214 Raw protein, g / kg 82 180 UFL / kg 0.9 0.9 Within 4 hours before attempt each day, with a cannula, he picks up a sample of a ruminal juice with each cow. This pre-expression period serves as a control. During the experiment, 0.1% of the dry powder containing 10% of the Bacillus cereus spores, C.I.P. strain, is added to the concentrated feed. 5832, per gram. The experiment lasts for 5 days. During the last two days of the experiment, each of the cows is removed from each cow. It is always taken around 8 o'clock in the morning, before feeding.

It becomes collected from the rum, filtered through gauze to remove feed particles.

Each sample of filtered juice from the rum is subjected to analyzes; Calculating / Cillus cereus strain C.I.P. 5832, by methodJ. Michard and L. Levesque, 1989, - pH adjustment by electronic pH meter (Schott brand) - <heavy volatile fatty acids, acetic acid, propionic acid, butyric acid, gas chromatography (Girdel chromatograph), - determination of ammonia by Kjeldahl method after precipitation trichloroacetic acid proteins, - determined by DAFA method according to J.V., Czerkawski, 1974. The results are shown in Table 5 below.

Tabulak 5 - Research on fistulated cows

Control Period i Research Research Period and Statistical Evidence (Analysis of Changes) 3. cereus, CIP5832 (CFU / ml) 45 C00-1 540 NS (x) pH 6.84-0.13 '6.90-0.10 Volatile Total Acids ( mmol / l) 77.18-11.36 79.80-12, 10 · NS Acetic acid (mmol / l) 53.50-6.67 54.15-7.47 N.S. Propionic Acid (mmol / L) 10.47-2.95 10.12 ^ 2.19 N.S. Butyric acid (mmol / l) 10.87-1.93 11.31-2.84 N.S. Ammonia (mg / 1) 57 i 27 83 i 15 S. DAPA (mg / l) 855 - 85 1,290 i 90 S. N.S. = inconclusive, p> 0.05 S. = conclusive p p <0.05 During the experimental period, the spores of Bacilluscereus, CoI.P. 5832, pH value, total volatile acid content and acetic acid, propionic acid and butyric acid content. In contrast, treatment increases the amount of ammonia and DAPA.

Therefore, these results are different from those reported in Japanese Patent Application 1977-8381.

A 50% increase in DAPA (1290 vs. 855 mg / L) cannot result from the administration of very small amounts of Bacilluscereus spores, C.I.P. In fact, the number of spores measured extremely small in relation to the numbers previously reported by the microbial population of the tale is 45,000 for the mammalian juice. This increase in DAPA means that it has increased the amount of the original microbial population in the rum. - ε - Example 2

An experiment on dairy cows of 20 crossed breeds of Frisonne x Holstein is divided into two equal groups.

Feeding consists of a basic daily dose supplemented with concentrated feed. The basic daily dose consists of the following components: ray-grass, silage maize, grain, straw and urea.It is served daily to each cow in the amount of 17 kg of dry matter, 15,31UFL (Unites Fourrageres Lait - 1 435 g) PLIN ”and 1,428 g of“ PDIE ”(intestinal protein digestible units according to the French ruminant feeding system). This basic daily dose, supplied in the form of a composite, covers the basic production of 25 kg of milk per day.

The concentrated feed provides in kg of dry matter: 1 UFL, 142 g PLIN, 138 g PDIE, 80 g crude cellulose and 320 g amidone + sugars. The concentrated feed is distributed individually each time by an automatic distributor at a rate of 1 kg of feed per 2.5 kg of milk above the basic production (25 kg).

A group of ten cows receives the above-mentioned daily dose. This group is a control group.

In addition, the second group of 10 cows receives in addition to the indicated dose

Q 10 g per day of dry powder cow containing 10 revivable spores of Bacillus cereus, strain O.I.P. 5832, in a gramophone. This addition allows an average of 60,000 to 80,000 spores to be recovered in a 1 ml tonic juice.

Cows are twice a day. The amount of milk produced by each cow is measured automatically at each milking. The butter content and the protein content of the milk are also checked.

The experiment is performed for three weeks. The results are shown in Table 6 below

Control Group Bacillus Statistical Proof (Change Analysis) Milking, kg / day 34.81 37.82 ς (x) Fat Content,% 3.86 3.83 N.S. Protein content,% 2.87 2.85 N.S. N.S. = inconclusive, p> 0.05S. = conclusive, p ^ 0.05

The results show that the spores of Bacillus cereus, strain C.I.P. 5832, increase milk production without changing its composition. Effects of Bacillus cereus, strain C.I.P. 5832 are therefore different from the results of European Patent Application No. 88108549 * 2, which are obtained using the microorganisms Lactobacillus, Pedicoccus, Streptococcus, Leucono-stoc, Bacillus subtilis and Bacillus toyoi. Example 3

An attempt on young bulls to produce meat * 28 bulls of the Salers breed is divided into two equal groups.

The individual daily feeding consists of the following two components:. 5 kg of concentrated feed containing 870 g in 1 kg of dry matter, 187 g of crude protein, 109 g of crude cellulose, 120 g of amidone, 6.1 g of calcium and 5.9 g of phosphorus.

A group of 14 control bulls is administered alone, this dose.

The second group of 14 bulls were fed with the same dose, but Bacillus cereus spores, C.I.P. 5832, at a rate of 400,000 resuscitating spores per 1 g feed. This addition is calculated in such a way that about 30,000 revivable spores are obtained in 1 ml of Bechterberry.

The animals are marked with a ring number in the ear. Each piece of 10 cattle

These are considered individually before and after the experiment. 53 days of the experiment, the following results are obtained, in Table 7 ·

Table 7 - Young bulls attempt

Control Group Bacillus Statistical Compliance (Change Analysis) Live Weight, kg N.S. initial 312.9 312.2 final 397.3 402.7 S. Total weight gain, kg 84.4 90.5 (+7.2%) S. Average daily weight gain, g 1,339 1,436 (+7 , 2%) 3. Consumption of dry matter kg / day of ensiled maize 4.32 3.59 NS 4.35 4.35 N.S. a total of 8.67 7.94 N.S. Consumption Index 6.42 5.93 (-7.6%) S.N.S. inconclusive, p £ 0.05S. significant, p <0.05

Thus, these results demonstrate that the reversible spores of Bacillus cereus, strain C.I.P. 5832, are capable of improving the zootechnical performance of young bulls fed on meat, ie weight gain and consumption index (consumption index = kg of dry matter used per kg weight gain).

Claims (6)

7 ^ 2. 11 Α Α ϊ Ε Κ Τ É V Ο Ο Υ Υ Υ Υ CLAIMS 1. A method for increasing the microbial population in goat-chewing animals, characterized in that Bacilluscereus bacteria, C.I.P. 5832. 2. A method according to claim 1, characterized in that bacteria are used which are in part in the form of revivable spores. 3. A method of increasing the zootechnical performance of ruminant animals reared for meat, characterized in that the content of the microbial population in their meat is increased as described in any one of claims 1 or 2. A feed additive for use in the method of any one of claims 1 to 3. 5. Feeding feed for ruminant animals, comprising a feed additive according to claim 4. 6. The use of Bacillus cereus bacteria, strain C.I.P.5832 to increase the microbial population in the ruminant animal of a ruminant animal. tupuj e