RU2371190C2 - Medicine for treatment of gastrointestinal tract diseases in chickens - Google Patents

Abstract

FIELD: veterinary.

SUBSTANCE: medicine contains symbiotic system of strains of lactic acid bacteria Lactobacillus acidophilus D-75 and D-76 of medicine "Vitaflor", enrofloxacin and twin-80 with following ingredients ratio (% wt) symbiotic microorganisms of "Vitaflor" medicine - 10-20; enrofloxacin - 75-85; twin-80 - 1-2.

EFFECT: medicine reduces murrain of young birds and favours increase of day average additional weight on 10-15%.

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Description

The invention relates to medicine, namely to preparations for the treatment of diseases of the gastrointestinal tract (GIT) based on probiotics (living microorganisms or substances of microbial origin, which, with the natural method of administration, have a positive effect on the physiological, biochemical and immune responses of the host through stabilization and optimization functions of its normal microflora) and can be used in veterinary medicine, medicine, food industry and related industries.

To solve similar problems, it is currently proposed to administer orally the preparations containing individual colonies of microorganisms, in particular biological preparations based on lactic acid microorganisms (EP 0097484, 1984, class A23C 1/05: EP 0043962, 1980, class A23C 19/086 : US 4,289,888, 1979, CL A23C 9/123). Preparations, as a rule, contain bacterial cells, residues of culture fluid (QOL), fats, proteins, lactose, gum and other additives. The composition of the compositions is determined by the characteristics of the microorganisms used and the nature of the task.

Known therapeutic and prophylactic biological product Bactisporin for the treatment of a wide range of diseases, containing freeze-dried biomass of the strain Bacillus subtilis 3H and a protective medium based on sucrose-gelatin mixture or lactose (RU No. 2130316, A61K 35/66, 1999). However, Bactisporin is not effective against a number of pathogens of acute intestinal infections, for example, caused by bacteria of the genus Pseudomonas spp., Streptococcus spp., Enterococcus spp.

Known drug Biosporin for the prevention and treatment of human gastrointestinal diseases, containing living cells of strains of Bacillus subtilis 3 (VKPM No. B-2335) and Bacillus licheniformis 31 (VKPM No. B-2336), as well as a filler (RU No. 1722502, A61K 35 / 74, 1992). Biosporin is characterized by high antagonistic activity to a wide range of pathogenic and conditionally pathogenic microorganisms, however, the strains included in its composition are sensitive to antibiotics, therefore, the drug is not recommended for use in treatment with antibiotics.

Known drugs based on acidophilic microorganisms (RU 2064269, class A23C 9/12, 1994; SU 1227145, class A23C 9/123, 1983; RU 2080795, class A23C 9/12, 1997) containing bacterial cells and cultural residues liquids (QL) with some additives that improve its performance (gelatin, sucrose, skim milk), as well as water.

The disadvantage of such drugs is their relatively low biological activity associated with the process of deactivation of living microorganisms during their passage through the upper gastrointestinal tract (stomach, duodenum), as well as the lack of stability of their biological properties during long-term storage, which leads to a decrease in the effectiveness of therapeutic and preventive procedures .

The closest in technical essence to the claimed drug is the drug "Vitaflor" (US Pat. RF №2169573, 2001), which is a symbiotic culture of lactic acid bacteria, including Lactobacillus acidophilus D-75, and the strain Lactobacillus acidophilus D-76, deposited in the collection of the All-Russian Research Institute of Agricultural Chemistry on July 16, 1997, with a ratio of microorganisms of these strains from 4: 1 to 1: 4 and with living cells from 1 to 10 billion cells per 1 g of the preparation.

The drug "Vitaflor" helps to adapt the body to adverse external influences and is used, in particular, for the treatment of gastric maltlimph (US Pat. RF No. 2269353, 2006), as well as in the manufacture of various food products (US Pat. RF No. 2202217, 2003; US Pat. No. 2178975, 2002). The drug is not inactivated by the components of the gastric juice and its therapeutic effect persists for at least 1 hour. The use of a symbiotic system as a probiotic contributes to the displacement of pathogenic microorganisms from the gastric tract and thereby to the patient’s recovery.

The disadvantage of Vitaflora is its relatively low anti-infection activity, as a result of which it is necessary to use long (2-3 months) courses of drug administration, which makes it unpromising for use in industrial poultry farming.

The task facing the authors was to create, on the basis of Vitaflora, a more effective drug with a stronger and faster therapeutic effect against a wide range of infectious diseases.

The technical problem was solved in the creation of the drug, which along with the probiotic included a composition containing an antibiotic, while the nature, concentration and conditions of administration do not cause deactivation of bacterial strains. Enrofloxacin was used as an antibiotic, which has a wide antibacterial spectrum against gram-positive and gram-negative bacteria and mycoplasmas: Mycoplasma gallisepticum, Mycoplasma meleagridis, Ecsherichia coli, Salmonella spp., Haemo-philus spp., Pasterella spplipplipptlethipplipptlethrl. ., Staphylococcus aureus., Streptococcus spp. etc.

The technical result was achieved as a result of using a preparation containing the symbiotic microorganisms of the Vitaflor preparation, which additionally included enrofloxacin and a surface-active substance (surfactant) - tween-80 in the following ratio of ingredients (wt.%):

symbiotic microorganisms of the drug "Vitaflor" - 10-20;

enrofloxacin - 75-85;

tween-80 - 1-2.

A feature of the drug is the use of enrofloxacin in a water-soluble form, which, as it turned out as a result of the experiments, practically does not negatively affect this symbiotic system, while the effect of the antibiotic on pathogenic microorganisms is mainly preserved. In particular, enrofloxacin retains antibacterial action against microorganisms such as Mycoplasma gallisepticum, Mycoplasma meleagridis, Ecsherichia coli, Salmonella spp., Campylobacter spp., Haemophilus spp., Pasterella spp., Brodetella spp., Erysiplelothrix rhusiopathiac Kibsiella spp., Staphylo-coccus aureus., Streptococcus spp. and a number of others.

The use of enrofloxacin in a concentration of more than 85% leads to inhibition of the development of lactic acid symbionts, at a concentration of less than 75% its anti-infectious effect is weakened.

Using a concentration of Vitaflor of less than 10% of the mass does not allow a stable population of microorganisms to be obtained in the body, which reduces the duration of the positive effect of the drug and leads to the need for additional administration of a probiotic. Using it in higher concentrations reduces the anti-infectious effect due to a decrease in the content of the antibiotic.

The choice of the concentration of tween-80 is determined by the concentration of the antibiotic and the characteristics of the solution used. The concentration of surfactants should provide a complete translation of the antibiotic into a water-soluble form. Excessive amounts of tween-80 do not contribute to improving the properties of the drug.

The optimal content of components in the preparation was Vitaflor - 16%; tween-80 - 1.6%, enrofloxacin - 82.4%. The main studies were conducted on this composition of the drug, which received the code name "Lactofluorine."

The drug for therapeutic purposes is used, as a rule, in the form of a 5-10% aqueous solution for watering birds. At the same time, along with the therapeutic and prophylactic effect, an increase of 10-15% in the average daily gain in live weight of the bird was noted.

The preparation is a homogeneous suspension of yellowish-white color, without the presence of mold and lumps, with a sour-milk smell. The selected dose is determined, on the one hand, by the need to administer sufficiently large amounts of the active principle and takes into account the recommended doses of antibiotic administration into the body (daily prophylactic and therapeutic dose of enrofloxacin 3-5 mg / kg of bird weight for 3 days for colibacteriosis and 5 days for salmonellosis) and, on the other hand, a higher concentration of the drug gives the water a specific flavor, making it difficult for birds to drink.

The drug is obtained by mixing and thoroughly mixing the commercially available dry ingredients included in it. Before use, the drug is dissolved in a predetermined amount of water with stirring.

Veterinary drug Lactofluorokhin showed efficacy in the prevention and treatment of acute intestinal infectious diseases (salmonellosis, colibacteriosis, hemorrhagic enteritis, campylobacteriosis, mycoplasmosis), as well as complications after getting sick with infectious diseases (flu, NB, ILT, IB and chronic infectious bursal disease) caused by feeding affected feed with molds, mycotoxins, and enteritis of various etiologies. It can be recommended as an anti-stress factor when switching from one type of feeding to another.

Example 1. The effect of the composition and dosage of the composition on the intestinal microbiocenosis of chickens in experimental colibacteriosis.

To determine the effect of the composition on the intestinal microbiocenosis and the choice of the optimal formulation and dose, experiments were conducted under vivarium conditions.

In experimental studies broiler chickens of the cross Baros were used. Birds were kept in vivarium, in cell batteries such as BKM-3B. Feeding was carried out according to the “Recommendations on the normative feeding of farm poultry”, Ministry of Agriculture of the USSR, 1983. Chickens were delivered to the vivarium by road 6-8 hours after sampling.

The chickens of the experimental and 1st control groups were infected with E. coli culture at a dose of ELD ₅₀ (about 500 million microbial bodies per 1 head). Before infection of the chickens, the infectious dose of the pathogen (LD ₅₀ ) was determined on birds of the same age.

The chickens of the 1st experimental group were daily fed 10% lactofluorine solution, composition: Vitoflor preparation - 10%, enrofloxacin - 88%, Tween-80 - 2%) at a dose of 10 ⁶ microbial bodies per head (based on the concentration of lactic acid bacteria in day, for 3 days, starting from the daily age, according to the scheme outlined below.

The chickens of the 2nd experimental group were daily fed a 5% solution of lactofluorine, composition: Vitoflor - 20%, Enrofloxacin - 79%, Tween-80 - 1% at a dose of 10 ⁷ microbial bodies per head per day (based on the concentration of lactic acid bacteria ) once a day, starting from the daily age, according to the scheme described below.

The chickens of the 3rd experimental group were daily fed 8% lactofluorine solution, composition: Vitoflor - 16%, Enrofloxacin - 72.4%, Tween-80 - 1.6% at a dose of 10 ⁶ microbial bodies per head per day (based on from the concentration of lactic acid bacteria) once a day, starting from the daily age, according to the scheme described below.

The chickens of the 4th experimental group were daily fed 8% lactofluorine solution, composition: Vitoflor preparation - 16%, enrofloxacin - 72.4%, Tween-80 - 1.6% at a dose of 10 ⁷ microbial bodies per head per day (based on from the concentration of lactic acid bacteria) once a day, starting from the daily age, according to the scheme described below.

The control groups were salted with saline.

The scheme of lactofluorine quenching consisted of the following: the chickens were kept for 2 hours without water, then the preparation of the solution was dissolved in 10 ml of boiled water, and the control group chickens received 10 ml of physiological saline. After 45-60 minutes, drinking water was poured into the drinkers. To determine the quantitative and qualitative composition of the microflora of feces, chickens of the control and experimental groups were examined at the daily, 8-day and 15-day age. Data of intestinal microbiocenosis are presented in table 1.

The dynamics of intestinal microbiocenosis in chickens in the control groups without E. coli and in the experimental groups with E. coli, where Lactofluorine was used, does not differ significantly.

Thus, the use of veterinary Lactofluoroquinine in liquid form for chickens contained in the vivarium, starting from the age of one day, did not affect the dynamics of the formation of intestinal populations of lactobacilli, bifidobacteria, staphylococci, but slowed the colonization of the intestines by microorganisms of the enterobacteriaceae family. A decrease in the population level of these microorganisms in the intestine can reduce the likelihood of gastrointestinal diseases caused by intestinal microorganisms.

Doses of 10 ⁸ -10 ⁹ microbial cells per head per day are economically disadvantageous, because while significantly increasing the consumption of probiotic. Based on this, during further experiments, dosage options of 10 ⁶ and 10 ⁷ microorganisms per head per day were selected.

There were no differences between the experimental groups receiving lactofluoroquin in this concentration range at doses of 10 ⁷ and 10 ⁶ microbial bodies per head.

Example 2. In LLC “Breeding poultry farm Lebyazhye” for 3 months were tested the drug Lactofluorine in a dose of 10 ⁶ microorganisms per head per day in three groups of chickens with 10,000 heads in each group. The drug was administered to birds during evaporation. Chickens given enrofloxacin in equivalent doses were used as a control.

The average values for three batches of poultry, during the cultivation of which the veterinary Lactofluorhin was used, are presented in table 2, as a control, the averaged data for three batches of chickens that were used antibiotic are given.

table 2 The effect of the drug Lactofluorhin on the safety of chickens. Performance indicators Group of chickens, experience Group of chickens, control Group number one 2 3 one 2 3 Preservation,% 93.8 95.7 96.1 85,4 89.6 88.5 The average daily gain in live weight, g 50.9 52.1 53.1 40.8 42.5 43.3 Feed consumption 2.01 2.03 2.05 2.05 2.1 2.1

Example 3. The effectiveness of the use of the drug Lactofluorine against bird salmonellosis was shown in model experiments on the experimental infection of chickens of the broiler-6 cross with salmonella strain S.enteritidis.

For chickens from day old for 5 days the drug Lactofluorine with drinking water was used as a group method. At the age of 7 days, chickens from the experimental and control groups were infected with S.enteritidis at a dose of 3 · 10 ⁸ microbial cells. 6 days after infection, 5 chickens from the experimental and control groups were examined bacteriologically for the presence of salmonella in the internal organs, as well as for the clinical manifestation of the disease and the safety of the bird. Bacteriological studies were carried out according to the guidelines for laboratory diagnosis of human and animal salmonellosis, the detection of salmonella in feed, food and environmental objects.

The obtained results on the preventive effect of the drug are shown in table 3. The experiments showed that the drug lactofluorine in group use with drinking water for 5 days completely prevents not only the clinical manifestation of the disease, but also the formation of bacteriocarrier during alimentary infection of chickens at the age of 7 days at a dose from 10 ³ to 10 ⁶ salmonella.

In the birds of the experimental groups with the indicated infectious doses, salmonella from the internal organs was not isolated 6, 55 days after infection. It should be emphasized that against the background of the use of the drug lactofluorchine, Salmonella could not be found not only in the internal organs, but also on the mucosa of the blind processes of the intestine.

In the control groups, reactive and salmonellosis pathogens were identified even when birds were infected at a dose of 10 ³ -10 ⁴ microbial bodies both from the intestinal mucosa and from the internal organs.

At infectious doses from 10 ⁶ to 3 · 10 ⁸ microbial bodies of S.enteritidis, the drug prevents the clinical manifestation of the disease, provides 94-96% safety, reduces the degree of contamination of the internal organs.

To assess the effectiveness of the therapeutic effect in salmonellosis at the age of 7 days, the chickens of the experimental and control groups alimentarily infected S.enteritidis at a dose of 3 · 10 ⁸ microbial bodies.

The chickens in the experimental groups, the drug Lactofluorine was administered after 1, 6, 12 and 24 hours with drinking water as a group method after infection of the chickens with a pathogenic strain of S.enteritidis; physiological saline was used in the control groups.

Therapeutic efficacy was taken into account 6 days after infection. For this, 5 chickens were selected from the experimental and control groups and examined bacteriologically for the presence of salmonella in the internal organs, as well as the general condition of the bird, the clinical manifestations of the disease, safety, and pathological anatomical studies were evaluated. The results are shown in table 4.

From the data of the table it follows that lactofluorine, once administered, has a pronounced therapeutic effect up to 12 hours after infection of the chickens with the pathogenic strain of S.enteritidis. With the introduction of the drug 24 hours after infection of chickens, therapeutic effectiveness is reduced to 20%.

Thus, using the drug lactofluorine without using any other drugs, it is possible to solve the epizootological and epidemiological aspects of the problem of salmonellosis in industrial poultry farming.

Table 4 The level of therapeutic efficacy of the drug Lactofluorhin in infection of chickens S.enteritidis depending on the timing of administration of the drug The time of administration of drugs after infection (h) The number of chickens in the group (goal.) Ill (goal) Palo (goal) The presence of pathological changes (goal.) Vyd. cult. E.coli CHP% one
6 twenty
twenty 6
eleven 0
5 0
7 one
10 one hundred
75 12 twenty 19 10 13 19 fifty 24 twenty twenty 13 17 twenty 35 infected the control twenty twenty twenty twenty twenty 0 not infected the control twenty 0 twenty 0 0 -

The data on the practical use of the drug lactofluoroquin show that the claimed composition has a therapeutic and prophylactic effect on the bird in infectious diseases, and also helps to increase the average daily weight gain of chickens.

Claims (1)

Means for treating diseases of the gastrointestinal tract of chickens based on the symbiotic system of strains of lactic acid bacteria Lactobacillus acidophilus D-75 and D-76 of the drug "Vitaflor", characterized in that it additionally contains enrofloxacin and tween-80 in the following ratio of ingredients, wt.% :
symbiotic microorganisms of the drug "Vitaflor" 10-20 enrofloxacin 75-85 twin 80 1-2