RU2322995C1 - Biopreparation of veterinary indication and method for preventing gastro-intestinal diseases in neonatal calves due to its application - Google Patents

Abstract

FIELD: veterinary science.

SUBSTANCE: the present innovation deals with obtaining a biopreparation being of probiotic and immunomodulating activity. This biopreparation contains: probiotic "Baikal EM-1" 5.0-20.0 ml; tincture of "Alphyt-8" phyto-species 1.0-2.0 in 250.0 ml distilled water and decoction of marsh cinquefoil (Comarum palustre L) 0.2 g in 10.0 ml distilled water. For preventing gastro-intestinal diseases in neonatal calves this preparation should be introduced perorally for the period of 12-15 d starting since the second colostrums feeding. The innovation enables to efficiently apply the suggested biopreparation being simultaneously of probiotic and immunocorrecting properties, for inspecific prophylaxis of diseases in neonatal calves associated with immunodeficiencies which lead to disbacteriosis.

EFFECT: higher efficiency.

2 cl, 2 ex, 5 tbl

Description

The invention relates to veterinary medicine, relates to an immunoprobiotic preparation and can be used for the prevention and correction of functional disorders and diseases in newborn calves associated with exposure to stressful and depleting environmental factors.

It is known that preparations based on plants such as ginseng, Rhodiola rosea, zamani, onions, garlic, lemon, blackcurrant, Echinacea purpurea and others can stimulate the body's defenses and increase their overall resistance to diseases [1], but their effect is characterized unidirectionality (tonic, sedative, anti-inflammatory, epithelializing, antispasmodic, etc.), since the medicinal properties of medicinal plants depend on the presence of substances with a unique pharmacological effect, which contain It occurs only in medicinal plants of certain species or in related species of the botanical family and gives medicinal plants a strictly defined biological activity.

Known compositions of medicinal and prophylactic agents from plant materials that have a wide range of therapeutic and recreational effects, including action on factors of natural resistance, for example, tetraimmunophyte - a phytoextraction drug from St. John's wort, hay leaf, rose hips and beet root crops, which have an immunostimulating effect on the humoral immune answer [2], or phytopreparation, which is an aqueous extract from the herbs of St. John's wort perforated, thyme, peppermint and coltsfoot [3]. A common drawback of herbal preparations when used to prevent gastrointestinal diseases of newborn calves is their lack of effectiveness due to the physiological characteristics of their body.

At birth, the calf’s intestines are sterile, and colonization by microflora - bifidobacteria and lactobacilli, which make up 80-90% of the total number of intestinal microorganisms during normobiosis and maintain colonization resistance, occurs gradually by 20-25 days. In the first days of life, the intestine is populated by the microflora of the birth canal and the so-called shed microflora, which is mainly represented by enterobacteria, enterococci and other aerobes, which creates the conditions for the occurrence of gastrointestinal diseases with a symptom complex of diarrhea.

Probiotics are widely used in veterinary practice to reduce the influence of infectious agents, restore intestinal normobiosis, and metabolic breakdowns, for example, probiotics, for example, lactobacterin (lyophilized microbial mass of live antagonistically active lactobacilli Lb.fermenti and Lb.plantarum) [4], bifidobacterin (lyophilized active antagonist B. bifidum No. 1, No. 791 and LVA-3) [5], lactobifadol (a mixture of live acidophilus and bifidobacteria dried by a sorption method on a natural plant carrier), bifitrilak (lyophilis strains of Lactobacillus bulgaris, Lactobacillus acidophilus, Lactobacillus fermentum, Bifidobacterium bifidum) [6] and others. The main disadvantage of probiotics is that their use for the prevention of gastrointestinal diseases is not always accompanied by a positive effect due to the limited spectrum of antagonistic activity of the immune system and calves.

Probiotic preparations with active phytocomplexes are known, for example, the Velenol feed additive, including the Lactobifadol probiotic preparation and Spirulina VEL dietary supplement [7], a preparation for young farm animals, including the bifidobacterin probiotic preparation and oat cryopowder [8]. Active phytocomplexes that are part of well-known drugs are the components necessary for normal intestinal microflora (food substrates and metabolic regulators of their own intestinal microflora), but they do not have pronounced immunotropic properties.

As a preferred analog, we consider the probiotic preparation Baikal EM-1, which includes a symbiotic complex of 86 strains of microorganisms. The disadvantage of the drug is the need for a long course (30 days), however, the most dangerous periods of calf life are limited to the first 12-15 days [9].

The purpose of the invention is a biological product with probiotic and immunomodulating activity for non-specific prophylaxis of diseases of newborn calves associated with physiological dysbiosis and immunodeficiency.

This goal is achieved by the fact that the proposed biological product for oral use, with probiotic and immunocorrection properties, containing the probiotic preparation "Baikal EM-1" and additionally the herbal infusion Alfit-8 and decoction of marsh marsh (Comarum palustre L.), based on daily doses:

probiotic "Baikal EM-1" 5.0-20.0 ml Alfit-8 phytobrain infusion 1.0-2.0 g in 250.0 ml of distilled water decoction of saber-eared swamp (Comarum palustre L.) 0.2 g in 10.0 ml of distilled water,

which is administered orally for 12-15 days, starting with the second colostrum.

Alfit-8 phytosborry includes, as plant ingredients, the yarrow, the root of the frangipani, the grass of the tea penny, the leaf of the nettle, the grass of the shepherd’s bag and the grass of the motherwort five-bladed. This composition contains complexes of biologically active substances with anti-inflammatory and hemostatic properties, as well as a general strengthening effect.

Marsh cinquefoil (roots and rhizomes) contains essential oil, tannins, saponins, vitamin C, carotene, flavonoids and organic acids.

The plant components of the claimed biological product include prebiotic compounds (polysaccharides, amino acids, antioxidants, organic acids), as well as various substances with a pharmacological effect, in particular essential oils (have a broad spectrum antimicrobial effect, mainly on coccal microflora, the mechanism of which mainly consists in destruction bacterial membranes, followed by metabolic disturbance, aerobic respiration, synthesis processes), glycosides (increase nonspecific resistance to shea a wide range of adverse effects through energy optimization (improving transmembrane glucose transfer, incorporating lipids in energy metabolism, enhancing gluconeogenesis from metabolic waste), adaptive RNA and enzyme syntheses that are “needed” at the moment to enhance protection functions), flavonoids (possess, among other things, antioxidant and membrane stabilizing activity, which in combination with anti-inflammatory action provides protection of hepatocytes from the damaging infectious and toxic effects of various natural factors), polysaccharides, organic acids (maintain acid-base balance). Of particular interest from the point of view of the effect on immunity and natural resistance are trace elements, in particular Cu (the content of dicotyledonous nettle leaves, the shepherd’s grass in the leaves, the roots and rhizomes of the marsh cinquefoil - 43.3 μg / g), Mo (the content in the grass of the shepherd’s bag - 88.0 μg / g), Cr (content in the roots and rhizomes of the marsh cinquefoil - 30.4 μg / g), Ml (content in the leaves of the nettle nettle - 62.5 μg / g; roots in the rhizomes of the marsh cinquefoil - 50.3 μg / g), Zn (content in the roots and rhizomes of the marsh cinquefoil - 48.7 μg / g), Se (content in the leaves rapivy dioica - 22.9 ug / g), and vitamins A, E, F (yarrow), C (nettle, Potentilla marsh).

The biological product is obtained by mixing the calculated amounts of the probiotic Baikal-EM-1, the infusion of Alfit 8 phyto harvest and a decoction of the roots and rhizomes of the marsh cinquefoil. The infusion of Alfit 8 phytobarrow and a decoction of the roots and rhizomes of the marsh cinquefoil are prepared in the pharmacopeia method in the ratio of raw materials to the extractor 1: 250 and 1:50, respectively.

Example. Preparation of the Alfit-8 phytobox infusion: 1.0 g of the Alfit 8 phytobox was placed in an enameled dish, 250 ml of distilled water at room temperature was added, insisted on a boiling water bath for 15 minutes and cooled at room temperature. Preparation of a decoction of the roots and rhizomes of the marsh cinquefoil: 0.2 g of crushed roots and rhizomes of the marsh cinquefoil were placed in an enameled bowl, 10 ml of distilled water at room temperature was added, insisted on a boiling water bath for 30 minutes and cooled at room temperature, after which it was filtered through canvas, squeezing raw materials, added water to the required volume of extraction. 10 ml of the Baikal EM-1 probiotic was poured into 250 ml of Alfit 8 phytobarrow infusion, everything was mixed well, after which 10 ml of decoction of the roots and rhizomes of the marsh cinquefoil was added.

Table 1 shows the variants of the claimed biological product, including ratios with different amounts of the starting components. Each version of the biological product was tested on a separate group of newborn calves with 10 animals each, selected according to the principle of analogues, starting from the second feeding for 12 days. The first dose of a biological product was prescribed for 20-30 minutes before the second feeding. The control group calves were prescribed lactobacterin in accordance with the instructions for its use. Efficiency was determined on the basis of clinical examination data. The results are presented in table 2.

Table 1
Biological product options for newborn calves (based on the daily dose) Source ingredients Compositions The number of components per 1 dose one 2 3 four Probiotic "Baikal EM-1", ml 5.0 10.0 20,0 25.0 Alfit 8 phytobrain infusion, g in 250.0 ml of distilled water 0.5 1,0 2.0 2,5 Broth of Sabelnik swamp (roots and rhizomes), g in 10.0 ml of distilled water 0.2 0.2 0.2 0.2

table 2
The effectiveness of the biological product depending on the ratio of the starting components (based on the daily dose) Indicators Control (Lactobacterin) Compositions Source components: Amount per daily dose one 2 3 four Probiotic "Baikal EM-1", ml 5,0 10.0 20,0 25.0 Alfit 8 phytobrain infusion, g in 250.0 ml of distilled water 0.5 1,0 2.0 2,5 Decoction of marsh cinquefoil (roots and rhizomes), g in 10.0 ml of distilled water 0.2 0.2 0.2 0.2 Ill, goal. diagnosis 4 toxic dyspepsia 3 dyspepsia 2 dyspepsia 1 dyspepsia 1 dyspepsia It’s a goal. 2 one - - - Efficiency% 60.0 70.0 80.0 90.0 90.0 Preservation,% 80.0 90.0 100.0 100.0 100.0

As can be seen from table 2, the ratio of the initial components of the compositions 1 and 4 are impractical due to inadequacy of costs. The optimal ratio of the starting components of composition 3.

Example 1. The effect of the biological product on the immunonological status of newborn calves was determined in two series of experiments: 1 series - the economy, long-term adverse for the gastrointestinal diseases of newborn calves (incidence - 75-87%, death 17-22%), 2 series - the economy, relatively healthy in gastrointestinal diseases of calves (incidence - 19-22%, mortality - 1.0-1.5%). According to the principle of analogues, two groups of normally developed newborn calves were formed (body weight 30-32 kg, body length more than 80 cm, uniform coat throughout the body, presence of at least 6 incisors in the oral cavity, the time of the appearance of movement and sucking reflexes during 30 minutes after birth): experimental and control. The calves of the experimental group, starting from the second colostrum, were prescribed a biological product inside for 12 days in the following ratio of components (based on the daily dose): probiotic Baikal EM-1 - 20.0 ml; Alfit-8 phyto collection infusion - 2.0 g in 250.0 ml of distilled water; decoction of saber-eared marsh - 0.2 g in 10.0 ml of distilled water. The parameters of the immunological status of calves were studied at the end of the experiment according to modern tests generally accepted in biology. The results are presented in table 3.

When analyzing immunobiological parameters (table 3), the animals of the experimental groups established an increase in the basic indicators of cellular and humoral immunity. Of great practical importance was the increase in the number of immunoglobulins in the blood serum of experimental calves, which indicated a better digestibility of colostrum in newborns and an earlier start in the synthesis of their own immunoglobulins.

Table 3
The effect of the biological product on the immunological status of calves Indicators Scientific and production experience Episode 1 - a farm unsuccessful for gastrointestinal diseases of calves Episode 2 - a calf-free gastrointestinal illness Experienced group Control group Experienced group Control group T-lymphocytes,% 18.0 ± 1.2 15.6 ± 0.9 24.1 ± 1.1 17.6 ± 0.4 B-lymphocytes,% 8.5 ± 1.1 6.2 ± 1.1 7.5 ± 0.4 6.5 ± 0.3 Bactericidal activity,% 89.2 ± 1.2 54.3 ± 1.3 99.2 ± 1.2 67.6 ± 1.2 Lysozyme activity,% 4.2 ± 0.4 3.0 ± 0.3 4,5 ± 0,2 3.6 ± 0.3 Immunoglobulins, mg / ml G 16.62 ± 3.30 13.50 ± 0.82 16.8 ± 1.0 14.2 ± 1.2 M 1.9 ± 0.4 1.5 ± 0.3 2.10 ± 0.18 1.5 ± 0.18 Total protein, g% 5.9 ± 0.4 5.5 ± 0.2 6.2 ± 0.10 5.8 ± 0.3

Example 2. The effectiveness of prophylaxis using a biological product in accordance with the invention in comparison with known means (probiotics "Baikal EM-1, bifidobacterin in combination with oat cryopowder) was determined on newborn calves distributed according to the principle of analogues in groups. Schemes of prophylactic treatments: group 1 - starting from the second colostrum, within 15 days inside the biological product in accordance with the invention in the following ratio of components (based on the daily dose): probiotic "Baikal EM-1" - 20.0 ml; Alfit-8 phyto collection infusion - 2.0 g in 250.0 ml of distilled water; decoction of saber-eared marsh - 0.2 g in 10.0 ml of distilled water. Group 2 - starting from the second colostrum, within 15 days inside the probiotic "Baikal EM-1" in a dose of 20 ml; Group 3 - starting from the second colostrum, within 15 days inside bifidobacterin in combination with oat cryopowder (1:10) in a dose of 12 g. Control - lactobacterin in accordance with the instructions for its use. Efficiency was evaluated according to clinical examination and bacteriological studies. Feces were collected 2 and 15 days after birth. Differential diagnostic media were used to isolate microorganisms: Endo, blood agar, bismuth-sulfite-agar, selenite. The number of bifidobacteria and lactobacilli (the number of colony forming units (CFU / g) was determined by dilution on Blaurock and MPC-4. The results are presented in tables 4 and 5.

Table 4
The effectiveness of the biological product in accordance with the invention in comparison with known probiotic preparations Indicators Groups of animals 1-n = 24 2-n = 25 3-n = 24 4-n = 28 Applied drug Biological product in accordance with the invention Probiotic Baikal EM-1 Bifidobacterin oat cryopowder (1:10) Lactobacterin Ill, goal. one four 3 5 Preventive efficacy,% 95.8 84.0 87.5 82.1 The form of the disease,% light one 3 2 2 heavy one one 3 It’s a goal. 0 one one 2 Preservation,% one hundred 96 95.8 92.9 Duration of treatment, days 3,5 3,5 4.0 6.5 The average daily weight gain, g 637.0 ± 93.0 612.0 ± 25.0 629.0 ± 27.00 468.0 ± 67.0

Table 5
The results of bacteriological studies of feces of calves Groups Examination period (days after birth) Species composition of microflora of intestinal contents, CFU / g E.coli Proteus Klebsiela Citrobacter lactobacilli bifidobacteria 1 group - a biological product in accordance with 2 10 ⁵ -10 ⁷ + 10 ³ 10 ³ -10 ⁶ 10 ⁹ -10 ¹⁰ 10 ⁶ -10 ⁸ invention fifteen 10 ⁴ - - 10 ³ 10 ⁸ -10 ¹⁰ 10 ⁸ -10 ¹¹ Group 2 - Baikal EM-1 2 10 ⁵ -10 ⁷ + 10 ³ 10 ³ -10 ⁶ 10 ⁸ -10 ¹⁰ 10 ⁵ -10 ⁸ fifteen 10 ⁵ - - 10 ³ 10 ⁹ -10 ¹⁰ 10 ⁸ -10 ¹⁰ 3rd group - 2 10 ⁶ -10 ⁸ + 10 ³ 10 ³ -10 ⁶ 10 ⁷ -10 ⁹ 10 ⁶ -10 ⁷ bifidobacterin + oat cryopowder (1:10) fifteen 10 ⁶ + - 10 ³ 10 ⁸ -10 ¹⁰ 10 ⁷ -10 ⁹ The control 2 10 ⁶ -10 ⁷ + 10 ^s 10 ³ - 10 ⁶ fifteen 10 ⁹ -10 ¹⁰ + - - - -

Clinical data (table 4) confirmed the effectiveness of prophylaxis with the use of a biological product in accordance with the invention in comparison with known probiotic preparations (probiotic Baikal EM-1, bifidobacterin in combination with oat cryopowder and lactobacterin), which was manifested in a decrease in the incidence rate of 11.8, 8.3 and 13.7%, respectively, increasing safety to 100% (compared with 96, 95.8 and 92.9%, respectively) and the average daily increase in body weight. The biological product in accordance with the invention to a greater extent than known probiotic preparations stimulated the propagation of lactobacilli and bifidobacteria and inhibited the growth of conditionally pathogenic microflora, which provided colonization resistance, an immunostimulating effect and the prevention of the occurrence of gastrointestinal diseases.

The studies, including immunological monitoring indicators, confirmed that the claimed biological product has good bioavailability for the body of newborn calves, normalizes metabolic processes, increases immunity and natural resistance, which allows a wide range of its use with good tolerance and the absence of adverse reactions.

Information sources

1. Vinogradova T.A. et al. Practical herbal medicine, St. Petersburg, 1998.

2. L.A. Makaradze // Veterinary Medicine, 1999, No. 3.

3. RU No. 2250110C1, 2005.

4. Guidance on the use of dry lactobacterin in veterinary medicine, approved. GUV Gosagroprom of the USSR 05/19/1989.

5. Temporary guidance on the use of bifidobacterin dry in veterinary medicine, approved. GUV Gosagroprom of the USSR 05/19/1989.

6. No. 97101157 A, 1999.

7. Materials of the scientific and practical conference September 14-16, 2004. New in epizootology, diagnostics and prevention of infectious and non-infectious diseases of birds in industrial poultry farming ”S.-Petersburg-Lomonosov. 2004, p. 192-193.

8. RU No. 2231362 C2, 06/27/2004.

9. Interim guidance on the use of the probiotic Baikal EM-1 in veterinary medicine, approved. 12/22/2005.

Claims (2)

1. Biological product for veterinary use, including the probiotic "Baikal EM-1", characterized in that it additionally contains an infusion of phytobarget "Alfit-8" and a decoction of marsh marsh (Comarum palustre L.) based on the daily dose: probiotic "Baikal EM-1" - 5.0-20.0 ml, Alfit-8 phytobrain infusion - 1.0-2.0 g in 250.0 ml of distilled water and decoction of marsh cinquefoil (Comarum palustre L.) - 0.2 g in 10.0 ml of distilled water. 2. A method for the prevention of gastrointestinal diseases of newborn calves using a probiotic preparation, characterized in that the biologic preparation according to claim 1, which is administered orally for 12-15 days, starting from the second colostrum, is used as a probiotic preparation.