SU730274A3 - Method of enhancing growth of animals and birds - Google Patents

Description

The invention can be used in livestock and poultry when grown on m from productive livestock and poultry. Methods are known for enhancing the growth of animals and birds by using various preparations of antibiotics, for example, antibiotic X-5108 and its physiologically active salt {or spiramycin derivatives (2. Antibiotic preparations are introduced into the feed mixed with liquid or solid feed carriers, evenly distributing them in the mass of the feed or in the poyle. However, antibiotic preparations are still relatively expensive, and in some cases not always effective, which necessitates a further search for effective means of enhancing growth and productivity. Breeding livestock and poultry. The purpose of the invention is to increase the efficiency of the method of enhancing the growth of domestic animals and poultry. This is achieved by using a mixture of mitramccins produced by Streptomyces pilosus N CIB 11238, or formula chelate of mitramycin A formula, as an antibiotic drug ( Cd2H, 024) 2 Mg-2H2D OR chromomycin in an amount of 0.00025-0.005% by weight of liquid or solid feed carrier. The indicated antibiotic preparations are known and used in medical practice and are compounds from the group of the golden acid 3, 4. Connect and of gold acid group orally administered to animals, as a supplement to their diet, i.e. mixed with solid food dissolved in drinking water or for young animals, for example, pigs or calves dissolved in whole or skim milk. It is advisable to introduce the compound into omesis with the usual nutritionally balanced solid food, such an additional food product should contain from 0.0001 wt.% (1 g per 1 ton) to 0.005 wt.% (50 g per 1 ton) of the compound. Animals may feed on such supplementary food during the entire period of their growth or during only a part of their growth period, for example, at an early stage, and / or during the period preceding the slaughter. Attaining practice, an increase in growth rate allows animals to reach a market weight or a slaughter weight in a shorter period of time.

At optimal levels of growth acceleration, no toxic effects on animals were observed.

Preferred compounds of the malic acid group for use in the proposed method are sodium and magnesium salts, chelates or complexes of mitramycin A and a mixture of mitramycin.

The composition for use in the inventive method contains a compound of the gold acid group together with a liquid or solid edible non-toxic diluent or carrier. A suitable liquid diluent or carrier is, for example, drinking water, whole milk or skim milk.

A suitable solid, non-toxic diluent or carrier may be, for example, nutritionally balanced animal nutrition (typical standard broiler food made from milled grain and cereal by-products) and animal protein supplemented with vitamins and minerals, standard pig fattening feed or other common feeding products. An inert solid diluent or carrier may be a substance without nutritional value, for example, kaolin, talc, calcium carbonate, filling earth, attapulgite clay, ground oyster shells or hammer lime; in addition, it can be starch and lactose ..

The proposed composition may be in the form of an additional food product suitable for direct feeding of animals, in which case it will preferably contain 0.0001-0.005% by weight of a compound from the group of golden acid mixed with a normal nutritionally balanced animal food product. It can also be used as a concentrated admixture for dilution with ordinary animal food to produce additional food that is suitable for direct feeding of animals. In this case, such an impurity in the preferred embodiment will contain 0.005-50 weight. Compounds from the group of gold acid in a mixture with either a nutritionally balanced animal food, an inert solid diluent, such as ground lime, or starch or lactose. Such an impurity may be diluted in the usual manner, preferably in successively in two or more stages to ensure uniform mixing.

A method for producing a mixture of mitramycins is envisaged, including growing a mitramycin-derived strain of Streptomyces pilosus in an aqueous

a nutrient medium containing sources of assimilated carbon and nitrogen with mixing, under aerobic conditions at a temperature of 15-45 ° C, filtering the culture and isolating acid leachate from the filtrate in the usual ways.

A suitable mitramycin-producing strain of S. pilosus is a strain stored in the National Collection of Industrial Bacteria of the Ministry of Agriculture, Fisheries and Food. According to Torrey Research Station, 135 Ebbey Road, Aberdeen AB9 8DC, Scotland, under the designation N CIB 11238, this strain of S. pilosus has the following description:

A. Morphology (Yeast malt agar,, 8 days).

Aerial micelles exhibit simple branching. Spore chains are long (more than 10 spores) in spirals (spira). The surface of the spores (in the electron microscope) of the fleecy. B. View of the overgrowth after 12 days at 25-C on the following nutrient medium with:

Yeast Malt Agar.

Typical streptomycete colonies. Aerial micelles are white at first, becoming gray as the sporulation process (gray rows). Spore formation is abundant. The base of the micelle is brown. Diffusing pigment is missing.

Agar starch salts.

Aerial mycelium is pale gray, sporulation is weak. The base of buffalo micelles. Diffusing pigment is missing,

Glucose-aspartic agar.

Aerial micelles, gray, abnormal sporulation. The base of the micelle is gray. Diffusing pigment is missing.

Glycerol-aspartic agar.

Aerial mycelium gray, abnormal sporulation. The base of buffalo micelles. Diffusing pigment is missing.

Agar from oats nki.

Aerial micelles absent. The base of the micelle is white. Diffusing pigment is missing.

Saccharose nitrate agar.

Aerial micelles are pale gray. Arguing xopoaiee. The base of the micelle is the color of gray buff. Diffusion of pigment is missing.

Tyrosine agar.

Aerial micelles are pale gray. Arguing is good. The base of the micelle is gray. Diffusing pigment is missing.

Nutrient agar.

Aerial micelles are white. Slight sporulation. The base of buffalo micelles becomes almost black. Diffusing pigment is missing. Iron-peptone agar. Aerial micelles are gray-white. Arguing is bad. The base of the micelles is brown, becoming black. Slightly diffusing brown pigment (Melanin +, but weak). Melanoid pigment agar. Aerial micelles absent. Base of micelle dark brown Weak diffusing brown pigment (Melanin +, but weak). C. Physiological properties. Use as the sole carbon source (in Pridham / Gottlieb agar): glucose + xylose + arabinose + fructose + galactose +. raffinose mannitol + inositol + salicin ± (very weak) sg1haroza -t (weak) Hydrolysis: gelatin + starch + casein (milk agar) + Action in litmus milk. Peptonization and the formation of acid. Curing does not occur. Growth temperature Germination on yeast malt agar after 14 days at a temperature (° C): 2 5 10 15 20 25 30 27 45 5 1-1--vvt-- - For example, myogaty alcohol, for example, glycerol, is an acceptable source of assimilated carbon. , glucose, xylose, arabinose, rhamnose fructose, galactose, mannitol or inositol, and glycerol is the preferred carbon source. Glycerol is usually introduced into the medium in an amount of 2-5% (w / v). A suitable source of assimilated nitrogen is, for example, peptone, which is usually added in an amount of 0.5-2% (w / v). The nutrient medium additionally contains small amounts of other elements, for example phosphorus, in the form of, for example, primary acidic potassium phosphate or diammonium phosphate acid; magnesium in the form of, for example, magnesium sulfate or magnesium carbonate; sulfur in the form, for example, of the sulfuric salt of sulfuric acid; potassium in the form of kyosha chlorine, potassium carbonate, or potassium acid orthophosphate; and salt salts of elements such as copper, zinc, iron, manganese and molybdenum. A temperature of 32 ° C is suitable for fermentation, and the oxidized product can be extracted from the culture filtrate by conventional methods, for example, by extracting an organic water-immiscible organic solvent at a neutral or alkaline pH from the filtrate to remove unwanted neutral and alkaline products, oxidizing the aqueous phase and extraction of the target oxidized product nesmeshivakndims - vault with an organic solvent. An improved method of purification of the oxidized product is used, which includes its conversion into magnesium chelate or its salt by contacting the solution of the oxidized organic water-soluble organic solvent with magnesium silicate. Example I.S. Pilosus NCIB 11238 is grown as an oblique culture on four slopes in a tripodond agar agar medium, in medicine planes, for 7 days. Micellium and spores from the slope are washed off with sterile water {: (100 ml) into two-liter jars containing 1 liter of the medium of the following composition,% (wt./ab.): Glycerol5 Peptone Difco0.5 Kshtsk Carbonate 0.1 Primary acidic potassium orthophosphate 0.024 Magnesium sulfate .7HjO 0.02 Microelement concentrate0, 1 Deionized water up to 100 Composition of concentrate I microelements,% (wt./about.) S Iron sulfate-7H O0.1 Copper sulfate 5H.jp0.015 Zinc sulfate, 1 Manganese sulfate 4 H2O 0.01 Potassium molybdate 0.01 Deionized water to 100. Salts are added to a portion of deionized water, and the mixture is oxidized sufficiently volume of concentrated hydrochloric acid with deionized water. The zagam solution is sterilized in an autoclave at a pressure of 1.05 atm for 0.5 h. The four two-liter flasks thus prepared are shaken for 2 days on a circular shaker: The contents of the four flasks are then combined to graft a fermenter containing 80 l of medium of the following composition,% (weight, / v): Glycerol5.0 Calcium carbonate 0.1. Potassium primary acidic orthophosphate 0.024 magnesium sulfate, 02 Concentrate of trace elements (as described above) 0.1 Peptone Difco0.5 Deionized water to 100 Then the solution is sterilized in an autoclave at a pressure of 0.05 atm for 0.5 h. The contents of the fermenter are mixed at 32 ° C for. 45 hours using a stirrer with six flat blades rotating at 260 rpm and aerating at 40 liters per minute. Polypropylene glycol is added to inhibit foaming. The fermentation broth is filtered and the filtrate (, 3) is extracted with ethyl acetate (25 L), which is then discarded. The pH of the aqueous phase is adjusted to 3.5 with 5N. hydrochloric acid and extracted with ethyl acetate (2 X 25 L). The ethyl acetate extracts are combined, dried with sodium sulfate and filtered, and the filtrate is concentrated under reduced pressure to a volume of 1 l. The target product from the above-mentioned concentrate can be obtained by the following methods. The ethyl acetate concentrate is washed with a dilute solution of sodium bicarbonate (500 ml of water, a fairly saturated solution of sodium bicarbonate to obtain a pH between 9 and 10). The aqueous phase is separated and washed with chloroform (1 L), which is discarded. The pH of the aqueous phase is then adjusted to 3.5 with diluted hydrochloric acid and then washed with DIETHYL1LM ether (2 x 1 L which is discarded and extracted with this acetate (2 x 1 L). The ethyl acetate extracts are combined, dried with sodium sulfate and filtered; evaporated to dryness and the residue triturated with diethyl ether to give the crude mixture of mitramycins as a tan solid. The ethyl acetate concentrate is evaporated to dryness to a crude gum. The resin (I is dissolved in methanol and the solution is adsorbed on kizel hur (about 2 g). The solvent is evaporated and the dry material is placed on top of a diatomaceous silica gel column (40 X 6.5 cm) filled with chloroform. The column is eluted with chloroform and 15 ml fractions are collected. These fractions are analyzed by thin-layer chromatography on a silica gel I Not Mezck 254 chloroform (methanol) of 98% mureshin acid 84: 8: 8 by volume for the presence of the target mithramycin products and fractions 50-250 containing this product, eluted with the product eluted and evaporated to dryness to obtain the crude mixture of mithramycins as yellow brown th solid (1.9 g). Example 2. A crude mixture of mitramycins (3.4 g) of example 1 in an aqueous solution of methanol is placed on top of a column (28 x 5 cm) pre-impregnated with cross-linked dextran Sephadex G-25 and the column is eluted with water. Citramycin is passed down the column in greenish - a yellow, fast moving layer and collect the corresponding eluant fraction. BUT). The eluant is dried at a temperature below 0 ° C to obtain mixed mitramycins as a yellow diophilic solid (2.5 g). AT). The eluant is adjusted to pH 3.5 with dilute hydrochloric acid and extracted with ethyl acetate, and the extract is dried and evaporated to dryness to give mixed mitramycins as a yellow OUJKa (2.3 g), m.p. 169-172 ° C. The mixed mitramycins thus obtained are converted to the main component of magnesium chelate as follows: Florisil is the trade name of magnesium silicate. Florisil. (6 g) is added to a solution of mixed mitramycins (400 mg) in ethyl acetate, and ethyl acetate is then evaporated. the risil with adsorbed mitramycin mixed is placed on top of a column (400 cm X 2 cm) filled with florisil, and the column is successively eluted with ethyl acetate, 5% (v / v) methanol in ethyl acetate, 10% (v / v) methanol in ethyl acetate, 15% (v / v) methanol in ethyl acetate. Fractions III and IV are combined, the solvents are evaporated, and the residue is crystallized from methliol / ethyl acetate to give magnesium chelate as yellow crystals, mp. 210-217С (decomposition), L + 85 ° (from 0.5 in methanol). Found,%: C 56.0; H 7.2. Calculated for (Cd2H., 5O2i) Md X 2H20, C 56.0%; H 7.0%. Example 3. A solution of the crude mixture of mitramycins (from example 1) 250 mg is dissolved in a minimal amount of methanol and placed on top of a column of silica gel (25 x 2 cm) to obtain as a bottom phase a solution from a mixture of diluted volumes of chloroform, methanol and hydrate ammonium oxides. The same bottom is used for the elution of the calico and the main yellow freshness is collected. The solvent is evaporated to give a yellow powder, which is dissolved in water (10 ml), the pH of the solution is adjusted to 3.5 using diminished hydrochloric acid and extracted with ethyl acetate (2 x 15 ml). The ethyl acetate extracts are evaporated to dryness to give mixed mitramycins as a yellow powder (100 mg), so pl. 169-172 C. Example 4. Groups of 20 selected chickens at the age of 1 day are placed in small pens with flooring, filled with wood chips and equipped with automatic rollers. In 16 such pens of birds for 6 days they fed the nutritionally controlled control feed, and birds in other pens (four pens for the food being developed) were fed during the same period of time with the same main feed, to which the specified weight of mixed miters received in Example 2 or 3, or a known chicken growth accelerator, penicillin or nitrovin, as a positive control. At the end of the test period, the average weight of the keicdah of the bird in the control group and in each study group is determined, and the percentage weight gain for the kg of each test group compared to the control group is calculated. Get the results shown in the table. 1. Example 5. With additional testing, the procedure specified in Example 4 is repeated. In this case, the weight is expressed as the average total weight of the pen of 20 birds. In tab. 2 shows the results of the tests. Example 6. In a further growth study, the procedure of Example 5 was repeated, but in this case all the feeds were pelleted. The results of the study are shown in Table. Example 7. The procedure of the example is measured using mitramycins obtained by cultivation of S. argillaceus A.TC.C. 12956 in a known manner. The results of the study are given in Table. 4. Example 8. The procedure of the example is repeated using the magnesium chelate indicated in Example 2. The results are given in table. 5. Example 9. The procedure described in Example 4 is repeated with the magnesium chelate of mitramycin A, obtained in a known manner by fermentation of S. argillaceus A.T.C. 12956. Get the results given in table. 6. Example 10. The procedure described in example 4 is repeated, using 4 pens with 30 males in each for each food recipe and extending the duration of the experiment to 4 weeks. The results are given in table. 7. Example 11. Seventy-two pigs weighing about 20 kg are divided into 8 corresponding groups of investigations of 9 pigs by sex and live weight. Each study group is planted on one specific diet and each animal in each group is fed with individually appropriate food, precisely weighed in accordance with the norm. Daily live weight gain in grams (DLWG) and relative feed utilization (FCR) - the ratio (in kg) of feed consumed per kg of live weight gain is determined for each group at the end of the nine week test period. Get the results shown in the table. 8. Example 12, The procedure described in example 4 is repeated using chromomycin AJ, obtained from So Foods Industries Limited, Osaka, Japan, instead of mixed mitramycins. The results shown in Table. 9. Example 13. The crystalline magnesium chelate from example 2 (4.2 g) was dissolved in water (100 i-m) and the pli was adjusted to 3.0 by adding 0.1 n. hydrochloric acid. The oxidized solution is then extracted with ethyl acetate (2. × 100 ml), the extract is dried (Naj, SOj,) and the solvent is evaporated to give a yellow gum. After triturating with ether, the resin gives a yellow powder {2.5 g), which is crystallized from ethyl acetate and dried under high vacuum to obtain a material with mp. 173-6 ° C, which, by IR and NMR spectroscopy and high pressure liquid chromatography (HLPC), was identical with mitremycin A. Example 14: 1 L; of the fermentation broth obtained in Example 1, was shaken with a 20% chloride solution the pH was adjusted to 3.5 and the mixture was extracted with methyl ethyl ketone (2 x 300 ml). The extracts were combined and dried, concentrated to 10 ml and back extracted with a saturated sodium bicarbonate aqueous solution (2 ml). Then the aqueous phase is treated with methanol (5 ml) to obtain a fine precipitate of mitramycin sodium salt {30 mg). Impurities suitable for dilution with animal feed can be obtained by introducing 1, 5, 10, 25, or 50 g of mixed mitrs 1 mycin into standard feed of broiler chickens containing ground maize and fish meal with lysine, methionine, vitamins and minerals with finishing weights up to 500 f. Similarly, you can get other mixtures, replacing the mixed strains with any other compound from the group of the golden acid. Food suitable for direct feeding of poultry can be obtained by thoroughly mixing 500 g of pre-mix as described in Example 5 with 4.5 kg of standard feed of broiler chickens and then uniformly mixing the mixture thus obtained with 995 kg of standard feed of broiler chickens to obtain a product for feeding poultry containing 1.5, 10 25 or 50 g of a compound of group h, with a ton of acid per ton, depending on the concentration of the compound in the previously prepared mixture.

and

73027412

Table 1

at the level, 001.

The main CFU - control

Main feed + mixed mitramycins 25 ppm

Basis and oorm + mixed mitramycins 10 ppm

Main feed + mixed mitramycins 5 ppm

The main fodder is mixed mithramycins 2.5 ppm

Table

Table 3

37.2 35.3 18.9 10.4

feed - control41,30korm + penicillin 25 ppm47,9516,1

feed + nitrovin 10 ppm45,6710,6

feed magnesium chelate 10 ppm 49,6520,2

Magnesium Chelate 5 ppm 47.9716.2

 P 0.001 0.01.

The main root-mixed mitramycins 10 ppm

The main feed-mixed mitracin 5 ppm

- substantial with p 0.01 level - significant with p 0.001 level

Table

V

.Table

.X

1,782

360

Xx

1,789

360

KOFW Low Copper Main Feed - Control Main Feed + Petrovin 10 ppm Main Feed + Carbadox 20 ppm

Main feed + mixed mitra 1 mycin 25 ppm

Kopw with high content of honey. Main food - control Main food + nitrovin 10 ppm Main food + carbadox 20 ppm

Main feed + mixed mitramycin 25 ppm

 - at the level of p 0.05 - at the level of p 0.01

Table 8

2.56

2,381

2,292

2,201

2,349

2.308 2.184

2.245

Table

Claims (1)

Claim A method of enhancing the growth of animals and birds by administering antibiotic preparations with liquid or 50 solid feed carriers, characterized in that, in order to increase the efficiency of the method, a mixture of mitramycins produced by Streptomyces pilosus NCXB 11238 or magnesium chelate mitramycin A of the formula are used as an antibiotic preparation 75θ24 ^ 2. Mg '2H ₂ O, or chromomycin in an amount of 0.00025-0.005% by weight of the carrier.