Medicinal agent for prevention and treatment of bird viral influenza

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science. Medicinal agent applied for prevention and treatment of bird viral influenza is represented with polystibuim. Polystibium inhibits infectious activity of influenza virus.

EFFECT: development of effective method applied for treatment and prevention of bird influenza.

6 tbl, 6 ex

 

The invention relates to the field of veterinary medicine, more specifically to the determination means for the prevention and treatment of bird flu.

Highly contagious flu, no specific means of treatment and prevention, emergency variability and the ability of the pathogen long to persist in a variety of organisms contribute to the rapid spread of infection and complicate the implementation of veterinary measures for the prevention and eradication of the disease. The pathogen is spread primarily by airborne droplets, as well as through infected food and water and excretion of their activity. Influenza viruses (VG) type And infect various species of birds and mammals, including humans, horses, pigs, whales, seals, etc. They are characterized by high antigenic heterogeneity of the surface proteins hemagglutinin and neuraminidase (NA) and presented under item 15 subtypes and 9 NA. Natural hosts of influenza a viruses are birds of wetland complexes. From birds isolated viruses with all known combinations of surface proteins. Despite the antigenic heterogeneity of influenza viruses, only four subtypes: H1-H3 and H5 and two NA (N1-N2) circulated among the people. Flu outbreak among humans in Hong Kong in 1997, caused by avian influenza A/H5N1 virus showed EAP is pout, that overcoming interspecific barrier bird-man can occur under natural conditions. This was the first time direct transmission of the virus from poultry to humans. 18 gospitalizirovany 6 patients died, not detected the transmission of the virus from person to person. In 2003-2004, the number of infected people in different countries of South-East Asia has risen to 53, and 43 people died. The situation is complicated by the possibility of forming a new pathogenic reassortant between influenza viruses in birds (chicken in this case) and epidemic human influenza virus. In this regard, for arresting the spread of infection in the foreground is a complex of sanitary measures, based on the use of high level disinfectants and sanitize.

Known means for the prevention of respiratory diseases of agricultural animals and birds - iodinator, Iodinol and attritional containing iodine in a positive valence form. These also include aerosol drug use to combat infectious laryngotracheitis, colibacteriosis and aspergillosis in birds (SU 566420 [1]).

However, these tools are not effective against influenza viruses, have no residual effect. In addition, due to frequent hypersensitivity among people, including the people abolnikov livestock, the preparations of iodine, the service personnel during the spraying solutions, extracts the necessary exposure and active ventilation in the room is not allowed, which creates difficulties when carrying out disinfection measures.

You know the prophylaxis of influenza in animals and birds, including low molecular weight glycol, organic or boric acid and constituens and cationic surfactant, tween 80 or tween 20 or tween 60 (EN 2012332 [2]).

A disadvantage of the known tool is its relatively low efficiency, as well as the fact that the tool made according to the proposed formulation, used for disinfection of pig and poultry houses an atomized surface irrigation, as well as refills desbrieres (impregnation of descobriu), i.e. the animal and bird therapeutic or preventive effects are not exposed.

You know the remedy for the prevention and treatment of viral influenza birds, representing the birch bark extract, produced by LLC "Birch world" (EN 2283126 [3]).

The disadvantages of the known tools should be attributed to the relatively low efficiency, since, as follows from the description of the patent specification, the tests were carried out on mice when the total number of animals 30 copies, 10 in each group, and the survival rate at Maxim is through the drug dose was 70% (see table 6 in [3]).

Claimed as an invention for the prevention and treatment of viral influenza birds are aimed at expanding Arsenal of medicines.

This result is achieved in that the proposed use polysorin as a means for the prevention and treatment of viral influenza birds.

The drug polysorin (Latin name polysurminym) is inorganic ionoobmennym polymer. Refers to the pharmacological group detoxifying funds. The active substance is a white powder, odorless and tasteless, insoluble in water and alcohol. Included in the list of medicines of the Ministry of health from 31.12.1999, No. 472. Known purpose polyserena is the excretion of radionuclides (see V. Korzun. and other International Journal of Radiation Medicine, 1999, 2(2): 75-91 [4]). Binds radioactive isotopes of strontium and prevents their absorption from the digestive tract. Known application polyserena in the treatment of radiogenic goiter (EN 2076682 [5]), as well as for radiation treatment of pathologies caused by the penetration of radionuclides into the body (EN 2063229 [6]).

Use polyserena for the treatment of avian influenza is not known.

The essence of the invention is illustrated by examples of its implementation.

The study virucidal properties of the drug "Poleasure is".

Example 1. Used a highly pathogenic strain of influenza A (H5N1)isolated during the epidemic among poultry in the Novosibirsk region in July 2005. Strain actively reproducerea in cell cultures spew who used to study infectious properties of the virus. Also used culture media, serum, antibiotics, sterile plastic disposable utensils for the cultivation of the virus of bird flu.

The drug "Polysorin" was used in the form of finely ground powder. The suspension of the drug received distilled water or physiological solution. Deposition polyserena of the suspension was performed by centrifugation at 4°C for 10-15 min Obtained supernatant was used for the study of antiviral activity against influenza virus a (H5N1).

The results are presented in tables.

The data in table 1 indicate that the drug "Polysorin" in the form of a suspension in physiological saline or distilled water at different concentrations able to inhibit the infectious activity of influenza virus H5N1. Complete inhibition of infectious virus activity was observed when added to the suspension, containing the concentration polyserena of 3.12 mg / ml and exposure within 1 hour of influenza virus. Suspensiononline concentrations of 1.5 mg / ml was able to inhibit the infectious activity of influenza virus And 10,000 times or more for 30 min or 1 hour. The contents of the influenza a virus in physiological solution without drug in these conditions did not lead to a decrease in the infectious activity of cell cultures SPM (table 1).

Example 2. We studied the ability of physiological solution after deposition polyserena to inactivate infectious activity of influenza virus in birds. In this case, the received suspension polyserena in saline solution (0.3 mg / ml), and after the natural deposition of the drug during the day the liquid gently plastiroute. Adosados were selected and introduced into the vessel with the supernatant fluid of the influenza a virus for 1 hour, after which investigated residual infectivity. As can be seen from table 2, infectious activity was equal to less than 1,01 g, while in the control solution without drug titers of influenza virus was 7,01 g TCD 50/ml

In the second variant of the experiment, the suspension polyserena besieged by centrifugation at 2000 rpm for 5 min and examined the supernatant, adding virusologii material. As it turned out, clarified by centrifugation at 2000 rpm, and then at 16000 rpm and the supernatant was kept high ability to inactivate infectious properties of the virus of avian influenza H5N1. Actually there was complete inactivation of the virus at a sufficiently high titer of influenza virus in control - 6,01 g TCD 50/ml/p>

From the data of table 2 shows that resuspending in a small amount of saline sediment polyserena also able to inhibit the infectious activity of influenza virus within 1 hour.

The data of table 3 indicate that the supernatant obtained after precipitation polyserena by centrifugation, retains a high virucidal properties and conditions of its cultivation in 2 times, and breeding her 4 times led to the reduction of infectious virus activity almost 100,000 times compared to the control experiment.

Virucidal properties supernatant decreased during storage for 5 days at 4°C. Studies have shown that in these conditions the solution polyserena was able to inhibit the infectious activity of influenza virus And birds in 1000 times (see table 3).

As evidenced by the data presented in table 4, virucidal properties of physiological solution after precipitation of the drug "Polysorin" at a concentration of 3.0 mg / ml led to a complete inactivation of the infectious properties of the virus of influenza a virus subtype H5N1 within 10 min after exposure to the supernatant to virusologii material, infectious titer reached 4,01 g TCD 50/20 mm. Lower concentrations polyserena did not lead to the ability of physiological solution to suppress infectious properties of the virus of the bird flu.

Example 3. The study virucidal properties of the drug, obtained by passing water through the powder polyserena.

In table 5 presents the data obtained when the water passes through the cartridge, filled with polyserena every 5 liters of Water, passed through a cartridge containing the drug polysorin, has weak virucidal activity against influenza virus And birds. Only sample No. 6 was able for 30 min to reduce infectious activity of influenza virus a 100 times. However, in 7-th sample was found not to have virucidal properties of water.

Example 4. The study virucidal properties supernatant, depending on the retention period.

From the data presented in table 6, it is seen that the supernatant for 4 hours kept maximum virucidal activity against influenza virus. However, after 4 days virucidal properties it was down and she was able to reduce the infectious activity of the virus in 1000 times, and after 8 days of storage of the supernatant at 4°With, the titers of influenza virus And after 30 minutes of exposure in the supernatant was reduced 100 times. These data suggest a decrease virucidal properties supernatant containing polysorin as storage at 4°C.

Example 5. The study of acute and chronic toxicity polyserena on belimia.

The study of toxicity polyserena were performed on white mice weighing 18-20 g (5 goals for each tested dose). To determine the acute toxicity of the preparation was injected into mice intramuscularly and orally, once daily, in doses of 250, 500, 750, 1000 and 2000 mg/kg, and chronic toxicity, three times, with an interval of three days, 250, 500 and 1000 mg/kg of body weight. This account of their behavior, condition of coat and visible mucous membranes, towards the stern, mobility, breathing frequency, time of occurrence and the nature of the intoxication, the time of death or recovery. In addition, I gave blood for hematologic research, produced packrite for comparative evaluation of organs and tissues with the organs of the control group.

It was found that the drug, regardless of the dose and method of administration, had no acute or chronic toxicity. For the entire observation period all mice in both cases remained alive. However, the maximum dose of 1000 and 2000 mg/kg led to an increase in the number of leukocytes (15,8-16,7×10 mm), decreased hemoglobin and the number of red blood cells (5,6-6,0×10 mm) and lymphocytes (53,0-58,0%).

Example 6. Drug use polyserena in the focus on the bird flu.

In the Republican veterinary laboratory were delivered dead birds belonging to the private sector selenium is I Paraul karabudakhkenskiy district of the Republic of Dagestan. The descriptions of the veterinary surgeon operating the farm, the birds were observed following clinical signs: depression, lethargy, wheezing, cyanosis of the comb and earrings, swelling in the head and neck, diarrhea, hemorrhages on the skin of the extremities.

Pathological changes. When the autopsy found: rhinitis, sinusitis, conjunctivitis, edema of the subcutaneous tissue in the head and neck, hemorrhage in the trachea, skeletal muscle, mucous membranes of the digestive tract, especially in the intestine in the area of the bifurcation. Hemorrhages on the mucous badocco glandular stomach was observed in the form of multiple dots, spots and stripes.

Of bodies delivered birds by PCR was isolated genetic material of the virus of avian influenza (H5N1). For testing of the drug "Polysorin" sick bird in this individual farming, livestock were divided into experimental (600 heads) and control (50 goals) group.

Polysorin - crystalline powder white odourless. Insoluble in water. He was bred at the rate of 1 kg per 3 l of boiled, cooled water with the subsequent assertion of 4-5 hours. The supernatant was poured into the vessel brought it up to 3 l by podlivaya boiled, cooled water. To drink 1 l of supernatant was diluted in 100 l of tap water and poured into the prepared for this t is whether drinkers.

The case of poultry the next day after feeding the drug amounted to: in the experimental group 70 goals (12%)in the control - 12 goals (24%).

The next day after feeding in the Republican veterinary laboratory were shipped 10 goals sweepable birds of the experimental and control groups. At autopsy detected above the anatomic changes in the control and experimental group.

However, research in PCR control group of birds was selected genetic material of the virus of avian influenza (H5N1), while the bodies of birds of the experimental group genetic material of the influenza birds were not allocated.

Thus polysorin is highly effective for the treatment and prevention of viral influenza birds.

Table 1.
Virucidal activity suspension polyserena in various concentrations in relation to highly pathogenic variant of influenza virus H5N1
Infectious titers of H5N1 influenza virus to cell cultures spew (1 g TZG/ml)
The concentration of the drug (mg/ml)502512,56,253,121,5The control virus
Virus + with spense drug - 30 min0000<1,02,56,7
Virus + suspension of the drug 1 hour000002,06,7

Table 2.
The ability of the solution after precipitation of the drug "Polysorin" (0.3 mg / ml) to inactivate infectious properties of highly pathogenic variants of influenza virus H5N1
Options experience1 g TZG/mlOptions experience1 g TZG/ml
Adosados after natural deposition<1,0Suspension polyserena to centrifugation1,2
The precipitate0Adosados after centrifugation2000 rpm, 5 min0
16000 rpm 10 min0
Without medication7,0Saline without medication6,0

Table 3.
JV is the ability of different dilutions of the supernatant after precipitation of the drug "Polysorin" to reduce infectious activity of H5N1 influenza virus for 30 min and 1 hour (initial drug concentration of 3.0 mg / ml)
Razvednye supernatantInfectious titers of H5N1 influenza virus to cell cultures spew (1 g TZG/ml)
30 min60 min4 days
Solid solution005,0
1:200N
1:44,83,2N
1:87,05,0N
The control virus8,08,08,0

Table 4.
Virucidal activity of the supernatant liquid obtained after deposition polyserena on avian influenza A H5N1 (after 10 min of treatment)
The titer of the virus of avian influenza for cell cultures SPAWVirucidal activity of the supernatant liquid obtained after deposition polyserena concentration (mg/ml)
30,03,00,30,3:20,3:4the control virus
1 g TZG/ml003,04,0the 3.8 4,0

Table 5.
Virucidal action of the water passing through the cartridge containing polysorin, influenza a virus subtype H5N1
The titer of the virus of avian influenza for cell cultures SPAWVirucidal properties of water samples by passing through a cartridge filled with polyserena
Numbers of water samples treated with influenza virus And birds
1234567The control virus
1 g TZG/ml5,54,24,84,84,03,55,55,5

Table 6.
Dynamics of conservation virucidal properties supernatant obtained after precipitation of the drug "Polysorin" against influenza a virus subtype H5N1
The titer of the virus of avian influenza for cell cultures spew

1 g TZG/ml
The titers of influenza a virus subtype H5N1 after 30 minutes of exposure in the supernatant liquid is received after deposition of the drug "Polysorin" (the time of receipt of the supernatant solution)
4 hours4 days8 daysthe control virus
Adosados after deposition at 2000 rpm04,15,07,0
Adosados after deposition at 16000 rpm06,07,09,0

Application polyserena as a means for the prevention and treatment of viral influenza birds.



 

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