RU2399661C1 - Probiotic preparation for gummer body weight gain and feeding method - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: probiotic preparation is used for gummer body weight gain and made of B. subtilis strain "ТПИ" 13-07.06.21-DEP/VGNKI and B. Licheniformis strain "ТПИ" 11-07.06.22-DEP/VGNKI, deposited in the Russian National State Collection of Microbial Strains Used in Veterinary Science and Animal Industries (FGU VGNKI). The preparation represents mixed culture liquids containing microbial synthesis products of said strains, with microbial cell concentration of 5 billion m.c./cm³, in the volume ratio of said strains as 1:1. The mixed culture liquids contain mixed bolting and pea flour equal by weight taken in the ratio 1:1. The preparation is dried up to humidity (5±1)%.

EFFECT: gummer body weight gain.

3 cl, 1 dwg, 3 tbl, 2 ex

Description

The invention relates to biotechnology, in particular to livestock, namely pig farming, and can be used in the preparation of feed rations for young pigs.

Pig farming has always been considered not only economically profitable, but also highly profitable industry, which is largely determined by the important biological characteristics of the organism of pigs, which compares favorably with the main characteristics of farm animals of other species.

Healthy pigs have a high feed conversion and growth rate. When digestion is disturbed as a result of poor-quality feeding, animals quickly lose weight, because the absorption of nutrients and various components of the feed worsens, and as a result, the microbial biosynthesis of the gastrointestinal tract changes, especially in young pigs during weaning.

In the stomach of pigs, enzymatic breakdown of food occurs mainly. However, an important role in the digestion of pigs is played by the normal microflora of the gastrointestinal tract.

Bacteria of the genus Bacillus are representatives of the normal microbiocenosis of the gastrointestinal tract of pigs, and with their lack or absence dysbacteriosis can develop.

Weaning is the most critical stage in the life of a pig from the moment of its birth to slaughter. During weaning, piglets are left without breast milk and must survive on the feed that they are offered. Feed for weaned piglets should be fresh, contain milk protein or fishmeal in combination with vegetable protein sources. Probiotics in pig nutrition are microbiological feed additives containing live cultures of beneficial bacteria that have beneficial effects on the animal, restoring the microbiological balance in the intestines.

A known method of feeding pigs, which includes introducing into the main diet a feed additive from waste from the production of microbiological preparations, characterized in that as a feed additive use a centrate - a technological product - waste formed during the production of a probiotic from Bacillus bacilli containing bacterial cultures of Bacillus subtilis VKPMV -2335, Bacillus licheniformis VKPMV-2336 (RU C2 2236147, 09/20/2004). The use of a centrate as a feed additive in animal husbandry helps stabilize the normal intestinal microflora, displace pathogenic and conditionally pathogenic microorganisms, provides protection against a large group of viruses by stimulating local immunity, but does not sufficiently contribute to the increase in body weight of farm animals.

There is also known a method of feeding pigs, including the introduction into the diet of a feed additive containing glauconite and probiotic "Biosporin". (RU 2319391 C2, 03.20.2008). The use of glauconite as a feed additive helps to increase the defenses and natural resistance of the pig’s body, but does not contribute enough to the increase in live weight, in addition, a feed additive is produced from raw materials mined in a certain place (Kunashak district of the Chelyabinsk region), which affects its availability and cost.

The closest is the method of feeding piglets, including the introduction of dry beer grains into the feed, characterized in that before the dry beer grains are fed into the feed, they are enriched with the probiotic Probiocel with a content of Bacillus subtilis 8130 of at least 2.5 · 10 ⁷ CFU per 1 g of grains and make enriched pellet in the feed to piglets from 60 to 108 days of age 2 times a day in the amount of 6% by weight of the feed (RU 2284703 C1, 03.21.2005).

However, the disadvantage is the difficulty in preparing the preparation, since fresh beer pellet is a perishable product, therefore it can be used only near the place of production. Dry beer pellet is most convenient for transportation, however, its feed value decreases after drying to 20-25%.

The technical result of this invention is to increase the gain in live weight of weaned piglets through the use of a new probiotic preparation based on the Bacillus subtilis strain TPI 13-07.06.21-DEPT / VGNKI and the strain Bacillus licheniformis TPI 11-07.06.22-DEPT / VGNKI deposited in the All-Russian state collection of strains of microorganisms used in veterinary medicine and animal husbandry (FGU VGNKI). The preparation is a mixture of culture fluids of strains containing products of microbial synthesis of these strains with a concentration of microbial cells of 5 billion cubic meters / cm ³ in a volume ratio of 1: 1, into which a mixture of bran and pea flour, equal in weight, taken in a ratio of 1 was added : 1, the preparation is dried to a moisture content of (5 ± 1)%.

The specified technical result is also provided by the described method of feeding weaned piglets, including the introduction into the diet of the above probiotic preparation at the rate of 5-7.5 g per 1 head.

Typically, this probiotic preparation is administered to weaned piglets as part of a full feed.

For the preparation of the probiotic, B. subtilis strains TPI 13-07.06.21-DEP / VGNKI and B. licheniformis TPI 11-07.06.22-DEP / VGNKI deposited in the All-Russian State Collection of Microorganism Strains Used in Veterinary and Animal Husbandry (FGU VGNKI) are used. , (A.P. Limorenko, P.G. Vasiliev, A.V. Vishnyakov, T.N. Gryazneva "Study of the effect of the sowing dose on the main indicators of the native culture during the deep cultivation of B. subtilis TPI 13 and B. licheniformis TPI 11 in nutrient media based on hydrochloric acid hydrolyzate of soy flour ", journal Veterinary medicine, 2007, No. 1, p.9).

B. subtilis TPI 13 - gram-positive aerobic spore-forming bacilli producing catalase. On MPA and wort agar with the addition of 50 μg / ml kanamycin form matte colonies of flesh or grayish color. In smears from colonies grown after 24-48 hours on potato agar, rods 2.6 × 0.7 μm in size are found containing oval-shaped centrally located spores. Colonies of old crops may turn black. They have plasmid and chromosomal resistance to kanamycin (50 μg / ml). On the BCH, the culture forms a film. Urea does not hydrolyze. Gives a positive reaction of Voges-Proskauer. Grows in the presence of 7% NaCl. Hydrolyzes starch and casein, liquefies gelatin. Ferments glucose, arabinose, xylose, mannitol with the formation of acid without gas. It does not need amino acids and vitamins for growth. Not pathogenic. Sensitive to species-specific bacteriophages. Coagulase and lecithinase activity does not possess. Antagonistic activity (zones of growth inhibition of test cultures) in relation to S. sonnei is 13 ± 1 mm; S. typhimurimn - 14 ± 2 mm; S. aureus - 23 ± 2 mm; C. albicans - 20 ± 2 mm.

Bacillus licheniformis TPI 11 - gram-positive spore-forming optional aerobic bacilli. Are mobile. On MPA, wort agar form matte colonies of beige with a rough surface. In smears from colonies grown on MPA, after 18 h, rods 2.5 × 0.6 μm in size are found containing oval-shaped spores that are located centrally. On BCH, culture forms catalase. Urea does not hydrolyze. Gives a positive reaction of Voges-Proskauer. Grows in the presence of 7% NaCl. Hydrolyzes starch and casein, slowly dilutes gelatin. It ferments glucose, lactose and mannitol with the formation of acid, reduces nitrates, under anaerobic conditions, forming a gas from them. It has coagulase and leticinase activity. Sensitive to species-specific bacteriophages. Not pathogenic. Inhibits the growth of test strains of S. sonnei - 16 ± 2 mm; S. typhimurimn - 12 ± 2 mm; S. aureus - 26 ± 2 mm; C. albicans - 25 ± 2 mm.

A probiotic is prepared as follows.

The first technological stage of preparation is the cultivation of the initial cultures of B. subtilis bacilli TPI 13 and B. licheniformis TPI 11 in test tubes on mowed agar. Next, the seed is grown in a dense nutrient medium Gause, in biological mattresses with a volume of 20 dm ³ at a temperature of (37 ± 1) ° C for 5-7 days. Daily, starting from 3 days of cultivation, the typicality (heterogeneity) of growth and the level of spore formation of bacilli is determined by microscopic analysis of smears from biomass of bacilli stained by Ziehl-Nielsen. After 6-7 days of cultivation, B. subtilis TPI 13 and B. licheniformis TPI 11 evaluate the completeness of spore formation.

The second technological stage of preparation is the transfer of bacillus cultures to the KM-0.1 bioreactor, where seed is prepared in liquid media PS-4 for B. subtilis TPI 13 and PS-7 for B. licheniformis TPI 11, table 1.

Table 1 The composition of culture media used for industrial cultivation of the studied bacilli Nutrient components Substation No. 4 Substation No. 7 COOK, cm ³ / dm ³ 60 21 HA cm ³ / dm ³ 8 70 Glucose, g / dm ³ eleven 12 MnSO ₄ · 5H ₂ O, g / dm ³ 0.3 0.3 MgSO ₄ · 7H ₂ O, g / dm ³ 0.25 0.25 FeSO ₄ · 7H ₂ O, g / dm ³ 0.01 0.01 CaCl ₂ · 2H ₂ O, g / dm ³ 0,046 0,046 NaCl, g / dm ³ 10.0 1,0

The cultivation parameters adopted for KM-0.1 (aeration coefficient 0.7 dm ³ / min, cultivation time 24 h for B. subtilis TPI 13 and 0.4 dm / min and 26 h for B. licheniformis TPI 11, respectively as well as a stirrer speed of 500 rpm) are sufficient to ensure the full growth and spore formation of bacilli.

Prepared seed crops of B. subtilis TPI 13 and B. lichemiformis TPI 11 should have the following indicators:

- the content of cells in the account in the Goryaev’s cell is not less than 15 billion / cm ³

- the content of living cells by the BK method is not less than 10 billion / cm ³ ;

- the content of mature spores is not less than 80%.

The next stage of the technological process is the cultivation of bacilli in the bioreactors BIOR-0,1. Seeds are introduced into the bioreactor at the rate of 1 million live spores per 1.0 cm ^{3 of} nutrient medium.

The parameters and modes of deep cultivation of bacilli are presented in table 2.

table 2 Parameters and modes of deep cultivation of bacilli Options and Modes B. subtilis TPI 13 B. lichemiformis TPI 11 Type of bioreactor BIOR-0,1 BIOR-0,1 PS Number 4 Number 7 Bioreactor fill factor 0.6 0.6 Temperature ° C 37 ± 1 37 ± 1 Pressure, MPa 0.10 ± 0.05 0.10 ± 0.05 Aeration coefficient, dm ³ / min 0.7 ± 0.1 0.4 ± 0.1 Mixer rotation frequency, rpm 530 320 Duration of cultivation, h 24 28 Defoamer E-1, dm ³ 0.8 0.5

The reason for the end of cultivation is the accumulation of biomass in the culture fluid of at least 5 billion cubic meters / cm ³ .

The next stage of production is the preparation of a mixture of culture fluids B. subtilis TPI 13 and B. lichemiformis TPI 11 in the homogenizer containing microbial synthesis products of bacilli and biomass of bacilli with a concentration of microbial cells of 5 billion MK / cm ³ in a volume ratio of strains 1: 1 . Then, an equal mass mixture of wheat bran (GOST 7169-66) and pea flour, taken in a 1: 1 ratio, is introduced into the homogenizer. Pea flour plays the role of a source of essential amino acids. Mixing is carried out at a stirrer speed of 1000 rpm.

The resulting mass is dried by the contact sorption method in an EVZ drying unit with a fluidized bed for 3 hours at a temperature of 40 ° C and 3 hours at a temperature of 60 ° C to a humidity of (5 ± 1)%.

The probiotic is packaged in plastic bags of 50 g each.

The invention is illustrated by the following examples:

Example 1. Obtaining a probiotic

1. Growing cultures of B. subtilis TPI 13 and B. licheniformis TPI 11 in test tubes on mowed agar.

2. The cultivation of seed in a dense nutrient medium Gause in biological matrices with a volume of 20 DM ³ at a temperature of (37 ± 1) ° C for 5-7 days.

3. The cultivation of seed in the bioreactor KM-0,1 in liquid media PS-4 for B. subtilis TPI 13 and PS-7 for B. licheniformis TPI 11 (aeration coefficient 0.7 dm ³ / min, cultivation time 24 h - for B. subtilis TPI 13 and 0.4 dm ³ / min and 26 h - for B. licheniformis TPI 11, respectively, the number of revolutions of the stirrer - 500 rpm).

4. Cultivation of bacilli in BIOR-0.1 bioreactors (aeration coefficient 0.7 dm ³ / min, cultivation time 24 hours for B. subtilis TPI 13 and 0.4 dm ³ / min and 28 hours for B. licheniformis TPI 11, respectively, the stirrer speed is 530 rpm for B. subtilis TPI 13 and 320 rpm for B. licheniformis TPI 11). Seeds are introduced into the bioreactor at the rate of 1 million live spores per 1.0 cm ^{3 of} nutrient medium.

5. Preparation in a homogenizer of a mixture of culture fluids B. subtilis TPI 13 and B. lichemiformis TPI 11 containing products of microbial synthesis of bacilli and biomass of bacilli in a ratio of the concentration of microbial cells 1: 1. Then, an equal mass mixture of wheat bran (GOST 7169-66) and pea flour, taken in a 1: 1 ratio, is introduced into the homogenizer. Mixing is carried out at a stirrer speed of 1000 rpm.

6. Drying the obtained mass by contact sorption method in an EVZ drying unit with a fluidized bed for 3 hours at a temperature of 40 ° C and 3 hours at a temperature of 60 ° C to a humidity of (5 ± 1)%.

Example 2

To feed weaned piglets, the probiotic is introduced into the diet at the rate of 5-7.5 g per 1 animal, which provides an effective increase in live weight gain in weaned piglets and is confirmed by the following example.

When using a probiotic for fattening pigs of the experimental groups, an additional increase in live weight of 9.2 kg per average head was obtained (table 3).

Table 3 Efficiency of using a probiotic for feeding weaned piglets Indicators Group of animals I experienced I counter. II experienced II counter. Dose KD-5 per 1 head, g 5,0 - 7.5 - The gain in live weight, kg 67.4 58.2 69.9 58.8 Feed consumed, kg 220.18 220.18 272.9 272.9

The increase in total live weight is shown in the drawing.

It was found that in the first experimental group of weaned piglets, the increase compared to the control is 9.2 kg, and in the second group - 11.1 kg.

Thus, the efficiency of using feed and the intensity of fattening weaned piglets was significantly increased when using a probiotic.

Claims (3)

1. A probiotic preparation intended to increase the gain in live weight of weaned piglets based on the B. subtilis strain TPI 13-07.06.21-DEP / VGNKI and the strain B. licheniformis TPI 11-07.06.22-DEP / VGNKI deposited in the All-Russian State collection of strains of microorganisms used in veterinary medicine and animal husbandry (FGU VGNKI), which is a mixture of culture fluids containing products of microbial synthesis of these strains, with a concentration of microbial cells of 5 billion cubic meters / cm ³ , in a volume ratio of these strains 1: 1 in which equal amount by weight of a mixture of pea bran and flour in the ratio 1: 1, dried to a moisture content (5 ± 1)%. 2. The method of feeding piglets, weaners, including the introduction into the diet of a probiotic preparation according to claim 1 at the rate of 5-7.5 g per 1 head. 3. The method according to claim 2, characterized in that the probiotic preparation is administered to weaned piglets as part of a full feed.

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