RU2429871C1 - Preparation to treat gastrointestinal diseases of calves and method of its application - Google Patents

Abstract

FIELD: veterinary science.

SUBSTANCE: preparation includes sterile culture broth produced by cultivation of fungus Trametes pubescens (strain 0663) on standard solid and fluid nutrient media with additionally added microcrystalline cellulose. The preparation is added into milk for bottle feeding in dose of 1.5-2.5 ml per 1 kg of animal live mass 1 time per day for 4-5 days.

EFFECT: preparation and method have high therapeutical efficiency in treatment of gastrointestinal disease of young farm animals.

3 cl, 12 tbl, 4 ex

Description

The invention relates to veterinary medicine, in particular to preparations and methods for treating gastrointestinal diseases of calves.

It is known to use xylotrophic basidiomycetes as producers of biologically active substances and to develop environmentally friendly non-waste technologies based on them to produce effective drugs for medicine and veterinary medicine, including those with antimicrobial, immunostimulating effects (Wasser SP Medicinal mushrooms as a source of antitumor and immunomodulating polisaccharids / / Appl. Microbiol. - 2002. - Vol.60. - p. 258-274; Belova HB Basidiomycetes - sources of biologically active substances // Raststnye resurs. - 1991. - Issue 2. - p. 8-18; Chkhenkeli VA Some aspects of study and use of biologically active substanc es of wood-rotting mushroom Trametes pubescens (Shumach.) Pilat. // Mat. of XV Congress of Europen Mycologists, St. Petersburg, 2007 - p. 185-186).

The closest solution adopted for the prototype is a method for treating colibacteriosis of calves by introducing Levan-2, obtained by the deep cultivation of fungi of the class Basidiomycetes of the genus Trametes of the species Trametes pubescens, in a drinkable milk in a dose of 30-90 ml (0.75-2.25 ml per 1 kg of live weight of the animal) per day until cure for 10-14 days (V. Chkhenkeli Biologically active substances of the basidiomycete Coriolus pubescens (Schum .: Fr.) Quel, and their use / RAAS. IF IEVS and DV. - Novosibirsk, 2006. - 288).

The disadvantage of the prototype is the rather low activity of the drug against bacteria of the Enterobacteriaceae family and the limited spectrum of action of the Levan-2 drug on other pathogens of gastrointestinal diseases of calves, as well as the need for long-term treatment.

The task of the invention is to expand the arsenal of tools and methods for treating gastrointestinal diseases of young farm animals.

The problem is solved in that a preparation for treating gastrointestinal diseases of calves, including a sterile culture fluid obtained by cultivating the fungus Trametes pubescens (strain 0663) on standard solid and liquid nutrient media according to the invention, a sterile culture fluid is obtained by cultivating the fungus Trametes pubescens ( strain 0663) on standard solid and liquid nutrient media with microcrystalline cellulose further included in them.

In addition, the problem is also solved by the fact that the preparation for the treatment of gastrointestinal diseases of calves according to the invention contains a solution of microcrystalline cellulose in an amount of 2.40 g / l.

The problem is also solved by the fact that in the method of treating gastrointestinal diseases of calves, including the introduction of the drug into milk for drinking, according to the invention, the drug is introduced in a dose of 1.5 -2.5 ml per 1 kg of live weight of the animal 1 time per day for 4-5 days.

Obtaining a drug for the treatment of gastrointestinal diseases of young cattle.

The initial step in obtaining the drug is to maintain the producer, i.e. culture of the fungus Trametes pubescens (strain 0663) on a solid nutrient medium - on wort agar. The seed is grown on Saburo standard solid nutrient medium, into which microcrystalline cellulose, which activates the producer’s natural metabolic activity, is preliminarily added. Next, seed is grown in a liquid nutrient medium. For seed and fermentation, a medium of the following composition (g / l) is used:

Potassium phosphate monosubstituted 1.88 Potassium chloride 0.94 Magnesium Sulphate 0.94 Sucrose 40,0 Microcrystalline cellulose 2.40 Peptone enzymatic pH 6.0 ± 0.2 12.50

The seed is grown in Erlenmeyer flasks in a shaker incubator at 220 rpm at a temperature of 28-30 ° C for 4 days. Seeds are introduced into the fermentation medium in an amount of 5-10% by volume.

Fermentation is carried out for 5-7 days at 220 rpm./min and a temperature of 28-30 ° C.

Thus, a change in the formulation during cultivation of the producer, namely, the introduction of an additional component, microcrystalline cellulose, into the nutrient medium for fermentation of the producer contributes to the activation of the producer's natural metabolic activity. This leads to an increase in the effectiveness of the drug and the expansion of the spectrum of its activity against pathogens of gastrointestinal diseases of calves.

Then the culture fluid is separated from the biomass of the culture mycelium by centrifugation for 10 min at 6000 rpm./min., Autoclave at 0.8 atm for 30 minutes

The resulting preparation is a sterile culture fluid obtained by deep cultivation of Trametes pubescens (strain 0663). The drug is a clear liquid with a yellow tint and a pleasant smell. The drug has low toxicity when introduced into the stomach and peritoneum in laboratory animals - white mice and rats. LD ₅₀ is 50,000 mg / kg and is close to the threshold in the acute experiment on the general toxic and dysbiotic effects. The locally irritating and allergic effect of the drug is poorly expressed. The content of salts of heavy metals in it is at the level of trace amounts. There are no residual quantities of pesticides, mycotoxins and nitrosoamines.

Examples of the use of the drug.

Example 1. For the optimal method of drug administration, only oral was adopted, since intramuscular and subcutaneous methods are unacceptable due to the strong reactogenicity of the drug, irritability, and painful reaction at the injection site.

To determine the therapeutic doses of the drug for gastrointestinal diseases of newborn calves with a symptom complex of diarrhea, mainly toxic dyspepsia and colibacteriosis, 72 calves of 1-10-day-old black-motley breed were selected, which were divided into 8 groups of 9 goals (Table 1).

Table 1 Groups of animals Dose, ml / kg of body weight Duration of treatment, days Therapeutic efficacy,% Preservation,% 1 experience 0.8 6-7 83.5 92.5 2 experience 1,5 4-5 87.3-100 100.0 3 experience 2.0 4-5 87.3-100 100.0 4 experience 2.5 4-5 87.3-100 100.0 5 experience 3.0 4-5 87.3-100 100.0 6 experience 4.0 4-5 87.3-100 100.0 7 Control with Levan-2 1,5 6-7 83.1 100.0 8 control (without treatment) - - - 57.5

The first 7 groups are experienced and the 7th, 8th group are control. Calves of 6 experimental groups were given the drug once a day in the following doses: 0.8; 1.5; 2.0; 2.5; 3.0; 4.0 ml per 1 kg of live weight of animals until recovery. The drug was added to milk before drinking. Group 7 calves received Levan-2 at a dose of 1.5 ml per 1 kg of live weight, which was added to milk before drinking. The calves were monitored for 10 days, the biochemical and morphological parameters of the blood were determined, the leukocyte formula was calculated, and the duration of the disease and safety were taken into account. Sick calves observed lethargy, lack of mobility, lack of appetite, increased intestinal motility, grayish, granular bowel movements with gas bubbles. During auscultation in the intestine, they listened to a fluid transfusion and other noises. On palpation of the walls of the abdomen, the calves experienced pain. The calves' hair was ruffled and moist, the skin of the perineum and in the tail area was smeared with liquid stool. A weak and slowed heart rate was determined, heart rate was weakened, slowed breathing, normal or subfebrile body temperature. During bacteriological examination of pathological material, E. coli (non-pathogenic for mice during their experimental infection) and other opportunistic microflora (Staphylococcus aureus, Pseudomonas aeruginosa, Proteus vulgaris, Streptococcus faecalis, etc.) were isolated. At autopsy, it was found that the mucous membranes were anemic-cyanotic, there were no hemorrhages, the spleen and lymph nodes were pale in color and not enlarged.

When treating calves with the drug in doses of 1.5-2.5 ml per 1 kg of live weight, calves recover after 4-5 days, while the treatment efficiency is 87.3-100.0%. With a decrease in the dose of the drug, its therapeutic effectiveness decreases to 83.5%, the duration of treatment increases to 6-7 days. Tables 2-4 show the results of biochemical, morphological studies of the blood of calves of all groups, as well as leukograms.

The data in tables 2-4 indicate that the optimal therapeutic doses of the drug should be considered doses of 1.5-2.5 ml per 1 kg of live weight of the animal.

When analyzing the biochemical parameters of calves (Table 3) treated with the drug, it was found that the level of total protein increased slightly (by 3.3–5.3%), other significant differences were not noted. The number of red blood cells in calves in groups 1-6 increased by 17.3%, hemoglobin - by 8.7-9.0% (Table 2). The leukocyte count in all experimental groups compared to the control group remained virtually unchanged (Table 2). Higher doses (> 2.5 ml per 1 kg of live weight) and lower (<1.5 ml per 1 kg of live weight) were not effective enough. The use of the drug before clinical recovery leads not only to a reduction in the duration of the disease, the exclusion of relapses, but also to further intensive growth and development of calves.

Example 2. To determine the therapeutic doses of the drug for colibacteriosis of calves, 72 calves of 10-15 days of age of black-motley breed were selected, which were divided into 8 groups of 9 goals (Table 5).

Table 5 Groups of animals Dose, ml / kg of body weight Duration of treatment, days Therapeutic efficacy,% Preservation,% 1 experience 0.8 6-7 82.1 93.5 2 experience 1,5 4-5 87.5-100.0 100.0 3 experience 2.0 4-5 87.5-100.0 100.0 4 experience 2.5 4-5 87.5-100.0 100.0 5 experience 3.0 4-5 87.5-100.0 100.0 6 experience 4.0 4-5 87.5-100.0 100.0 7 control (with Levan-2) 1,5 6-7 83.5 100.0 8 control (without treatment) - - - 67.8

The first 6 groups are experimental and 7, 8 groups are control. The calves of the experimental groups were given the drug in the following doses: 0.8; 1.5; 2.0; 2.5; 3.0; 4.0 ml per 1 kg of live weight of animals until recovery. The calves were monitored for 10 days, the biochemical and morphological parameters of the blood were determined, the leukocyte formula was calculated, and the duration of the disease and safety were taken into account. In calves, depression, loss of appetite, increased intestinal motility, fluid, beige colored feces were observed, bowel movements were up to 7 times a day, lethargy, unsteady gait, and shortness of breath were also observed. The body temperature was 39.0-40.1 ° C, the respiratory rate was 28-32. During bacteriological examination of feces, E. coli bacteria of serovars O ₉ and O ₂₆ pathogenic for white mice were isolated.

When treating calves with the drug in doses of 1.5-2.5 ml per 1 kg of live weight, calves recover after 4-5 days, while the treatment efficiency is 87.5-100.0%. With a decrease in the dose of the drug, its therapeutic effectiveness decreases to 82.1%, the duration of treatment increases to 6-7 days. Observation data are given in tables 6-8. The data in tables 6-8 indicate that the optimal therapeutic doses of the drug should be considered doses of 1.5-2.5 ml per 1 kg of live weight of the animal.

When analyzing the biochemical parameters of calves (Table 6) treated with the drug, it was found that the level of total protein increased by 16.7-17.4%, other significant differences were not noted. The number of red blood cells in calves in 2-4 groups increased by 11.5-14.6%, hemoglobin - by 23.5-34.7%) (Table 7). The content of leukocytes in all experimental groups compared with the control group decreased by 7.2-10.0%, mainly due to stab neutrophils and eosinophils (Table 8). Higher doses (> 2.5 ml per 1 kg of live weight) and lower (<1.5 ml per 1 kg of live weight) were not effective enough.

Example 3. To determine the therapeutic doses of the drug for calf salmonellosis, 72 calves of 80-90-day-old black-motley breed were selected, which were divided into 8 groups of 9 animals each (Table 9).

Table 9 Groups of animals Dose, ml / kg of body weight Duration of treatment, days Therapeutic efficacy,% Preservation,% 1 experience 0.8 6-7 84.5 100.0 2 experience 1,5 4-5 94.8-100.0 100.0 3 experience 2.0 4-5 94.8-100.0 100.0 4 experience 2.5 4-5 94.8-100.0 100.0 5 experience 3.0 4-5 94.8-100.0 100.0 6 experience 4.0 4-5 94.8-100.0 100.0 7 control with Levan-2 1,5 6-7 82.3 83.6 8 control without treatment 64.5

In sick calves, depression, loss of appetite was observed, the temperature at the onset of the disease was 39.9-40.9 ° C, and then in the majority it decreased to 39.1-39.5 ° C, feces were excreted with a high mucus content, from beige to dark color, watery, gruel-like consistency. During bacteriological examination of feces, cultures of Salmonella dublin and S.thyphymurium were isolated. The first 6 groups are experimental and 7, 8th groups are control. The calves of the experimental groups were given the drug in the following doses: 0.8; 1.5; 2.0; 2.5; 3.0; 4.0 ml per 1 kg of live weight of animals until recovery. The calves were monitored for 10 days, the biochemical and morphological parameters of the blood were determined, the leukocyte formula was calculated, and the duration of the disease and safety were taken into account. When treating calves in doses of 1.5-2.5 ml per 1 kg of live weight, calves recover after 4-5 days, while the treatment efficiency is 94.8-100.0%. With a decrease in the dose of the drug, its therapeutic effectiveness decreases to 84.5%, the duration of treatment increases to 6-7 days. Observation data are shown in tables 10-12.

The data in the tables indicate that the optimal therapeutic doses should be considered doses of 1.5-2.5 ml per 1 kg of live weight. When analyzing the biochemical parameters of calves treated with the drug, it was found that the level of total protein increased by 16.9-17.3% (Table 10). The number of red blood cells in calves in the 2nd-4th groups increased by 8.1-10.6%, hemoglobin - by 24.8-32.7% (Table 11). The content of leukocytes in all experimental groups compared with the control group decreased by 1.6–9.1% (Table 11), mainly due to stab neutrophils and eosinophils (Table 12). Higher doses (> 2.5 ml per 1 kg of live weight) and lower (<1.5 ml per 1 kg of live weight) were ineffective. This is primarily due to the fact that the drug in doses of less than 1.5 ml per 1 kg of live weight has a lower antimicrobial effect, and in higher doses it has an irritating effect on the mucous membrane of the gastrointestinal tract.

The rationale for the treatment of calves for four to five days is the average duration of gastrointestinal diseases of calves, i.e. the most optimal period when the disappearance of clinical signs and recovery of the animal.

The most optimal option for the treatment of gastrointestinal diseases of calves caused by bacteria of the Enterobacteriaceae family - colibacteriosis and calf salmonellosis - is the introduction of the drug into milk during drinking at doses of 1.5 to 2.5 ml per 1 kg of live weight of the animal once a day. When the drug is added to milk during drinking at a dose of 1.5-2.5 ml per 1 kg of calf live weight 1 time per day, the treatment duration for colibacteriosis and calf salmonellosis is 4-5 days, the treatment efficiency is 87.5-100%.

Example 4. Scientific and production experiments on the use of the drug for therapeutic purposes were carried out in two farms in the Irkutsk region of the Irkutsk region in coordination with the State Veterinary Service of the Irkutsk Region - at the MTF OAO Khomutovskoye and at the MTF GUP OPKh Baikal-Sibirskoye of the Russian Agricultural Academy at 167 calves.

The drug was administered orally with milk in doses of 1.5-2.5 ml per 1 kg of calf live weight according to the method described above. 60-100 ml / goal. with colibacteriosis and 120-200 ml / goal. with calf salmonellosis, because calves with colibacillosis were 10-15 days old, and calves with salmonellosis were 2.5-3 months old. The drug was administered once a day for 4-5 days. The therapeutic efficacy of the method is confirmed by studies: out of 167 calves that were treated with the drug, 151 animals recovered, which is 96.8% of the total. Of the total number of control animals (165 animals), which were treated according to the schemes generally accepted in farms, 126 animals, or 76.4%, recovered. Thus, on the basis of the experiments, positive results were obtained, indicating a high therapeutic effect of the drug in this method of its use.

Using the proposed drug and method of treatment allows you to expand the arsenal of complex drugs used to treat gastrointestinal diseases of calves caused by bacteria of the Enterobacteriaceae family.

Claims (3)

1. The preparation for the treatment of gastrointestinal diseases of calves, including sterile culture fluid obtained by cultivating the fungus Trametes pubescens (strain 0663) on standard solid and liquid nutrient media, characterized in that the sterile culture fluid is obtained by cultivating the fungus Trametes pubescens (strain 0663 ) on standard solid and liquid nutrient media with microcrystalline cellulose further included in them. 2. The drug for the treatment of gastrointestinal diseases of calves according to claim 1, characterized in that the solution of microcrystalline cellulose is added in an amount of 2.40 g / L. 3. A method of treating gastrointestinal diseases of calves, including the introduction of the drug into milk for drinking, characterized in that the drug according to claim 1 is administered in a dose of 1.5-2.5 ml per 1 kg of live weight of the animal 1 time per day 4-5 days.