FMD vaccine type a inactivated sorbed. Symptoms and treatment of foot and mouth disease in veterinary medicine

Acute infectious diseases can cause losses not only to large farms, but to small farmsteads. Therefore, it is important to recognize their symptoms in time and immediately begin treatment, especially since many of them are dangerous for people.

In this review, we will consider what foot-and-mouth disease is, what is its danger and how to deal with it.

Etiology of the disease

The causative agent of this disease is one of the smallest viruses - Dermaphilus, containing RNA. In spite of small size, has a high virulence (ability to infect).

Dermatotropism is pronounced - most often the disease begins with infection of skin areas or damaged mucous membranes. Spread through raw, meat and excretion products.

In addition to eating milk or undercooked meat for food, the contact route of infection is also dangerous for humans - veterinarians know that by touching the infected area, there is a risk of “picking up” such a disease. This also applies to slime particles. Fortunately, a person is not particularly susceptible to its action, which cannot be said about animals (especially artiodactyls).

There are 8 strains of this virus in total. In our conditions, the main types are A and O, other pathogens are rare.

Did you know? Last on this moment a major outbreak was recorded in the UK. In 2001, there were about a thousand foci of the disease- The epizootic was caused by strain O, which caused damage to the economy in the amount of $ 20 billion.

First symptoms

The incubation period of the virus is usually 2-4 days, but it is often prolonged. For example, in pigs it can last 7–8 days, and in pigs it can last up to 2–3 weeks. In this period visible reasons not to worry, although the disease progresses rapidly.

The alarms are:

• general weakness of the animal and loss of appetite;
• short-term increase in temperature;
• prolonged diarrhea;
• animals begin to fall on their forelimbs, limp (this is typical if foot and mouth disease has affected cattle);
• chewing gum lethargy;

• increased salivation;
• in some cases, the animal is unable to open its mouth.

Course of the disease

The disease occurs in acute form. In adult animals, it usually takes benign form, while malignant (it is also an atypical course) is extremely rare.
In different animals, the effect of the infection proceeds taking into account the characteristics of the species and breed.

Let's start with . After the late period (1–3 days, but sometimes from 7 to 20 days) has expired, the animal refuses to eat at all, the pulse quickens, and chewing gum stops. For 2-3 days active phase inside the lips, on the mucous membrane of the cheeks, tongue and edges of the jaw, aphthae (rash bubbles) appear.

In difficult cases, such formations are visible on the udder and between the hooves. The defeat of all limbs is rare, more often it is lameness on one pair of legs.

Important! After the sick animal has been taken out of the room, the inventory and the building itself must be treated with a disinfectant- 1% chloramine is quite strong.

12–24 hours after the appearance of the aphthae, they break, forming an erosion. At the same time, the temperature returns to normal, although salivation remains profuse, and foam is visible at the corners of the mouth. "Sores" heal in a week, but with complications, this process can take 13-20 days.
On the extremities you can see the same aphthae and swelling. They also burst and heal in 4–8 days. If the lesion is large in size, then there is a risk purulent diseases possibly even corneal detachment.

Aphthae in dairy cows inflame the teat canals, diseased quarters work with disorders. This is manifested in a change in the composition of milk: it becomes slimy and bitter. If the nipple canal is blocked by scabs, then it begins. At the same time, productivity is reduced to 60–75%, and it takes months to restore it.

A disease such as foot and mouth disease is especially dangerous for calves. They do not suffer from aphthae, but the disease is accompanied by severe disruption of the gastric tract. If help is late, a case may begin.

The "pure" type of virus stops its activity after 7-10 days. With background complications, the disease lasts much longer, up to a month. Basically, these are problems associated with the gastric and lactal highways.
With an atypical form, it is even more difficult: an animal that is recovering sharply “gives up”, refuses to eat, its hind limbs are paralyzed. Such a drop can occur 6-10 days after the onset of the disease. It hits the heart, and mortality, reaching up to 20-40%, in such cases is associated with stopping it.

Did you know? Foot-and-mouth disease has long annoyed livestock breeders: the first clinical description for animals back in 1546 was given by the doctor D. Frakastro. A similar picture for people was described much later by the Germans Frosch and Leffler, who proved in 1897 viral nature diseases.

The epidemic of foot and mouth disease is even more acute, not sparing the young. After 1-2 days of incubation, fever appears, appetite decreases. The limbs are affected, the pigs often limp (their hooves may even fall off). Aphthae are visible on the mammary glands, patches, and in oral cavity are rarely observed. The severe course of the disease is accompanied by bloody diarrhea and mucus, hemorrhages in the kidneys and lungs.

Adults get sick for a long time: from a week to 20-25 days. For piglets, foot-and-mouth disease is completely fatal (the case is at least 60%), the first two days of virus activity are considered the most dangerous.
Goats are a little easier. After 2–7 days of the latent period, the appetite disappears, the animal is in a fever, it begins to limp. At the same time, it is difficult for him to open his mouth, you can hear the gnashing of teeth.

Aftas appear on the hooves, mandible, lips and udder.

A sticky liquid flows from them. Goats are more resistant to FMD and complications are rare.

Full recovery occurs in two weeks.

Sheep after 2-3 days of the latent period limp, sometimes stop chewing and move little. The temperature can reach up to 41-41.5 °C.

In the case of them, aphthae are small, quickly burst and heal early. The affected area is the same: hoof gaps and corolla, gums, tongue and lips, upper jaw to the teeth.

Sheep recover after 10-12 days. Lambs often die due to complications such as septicemia (damage to tissues and the circulatory system).

Important! Patients before meals are given 0.1 g of anesthesin, which slightly smoothes the discomfort that occurs when eating.

But there is one caveat: in large flocks, the virus acts slowly and weakly, so its effect is not visible. Such a slow progress is very dangerous and can last 3-4 months or until it becomes acute.

Treatment of sick animals

Due to the variety of manifestations of the virus, the industry does not produce universal ones (with the exception of immunolactone, and even then it is not always suitable). Therefore, treatment is reduced to eliminating the symptoms.

The food is easily digestible: in summer it is grass, in winter they give soft hay or high-quality silage.

If foot and mouth disease in animals passes in the usual form, remedial measures come down to the following actions:

• The oral cavity is rinsed with weak solutions of potassium permanganate (0.1%) or furacilin (0.5%). Suitable and acetic acid at a concentration of 2%.
• With severe lesions of the oral mucosa, ointments based on copper, anesthesin or novocaine are taken. Fish oil is also helpful.

Did you know? Vaccination in its current form- in many respects the merit of Louis Pasteur. His legacy and diligence are impressive: in 1881, having made preparations against the Siberian bacillus, he was able to “neutralize” rabies with a vaccine four years later.

• Clean limbs daily. Hooves and corollas are treated with a composition of tar and fish oil in equal proportions. To fix the result, the animal is carried out through sawdust, which are soaked in tar. On large farms, formalin baths (5% solution) are made for the same purpose.

At severe forms diseases do the following manipulations:

• Severely affected areas of the limbs are smeared with iodine. Having cleared the hoof, the dead tissue is removed and the wound is cauterized with powder (½ of permanganate and streptocid), after which a bandage is applied.
• Aphthae on the udder are treated with novocaine-trypoflavin ointment based on petroleum jelly. They also interfere with it (15% of the volume). Synthomycin ointment also helps.
• If the complication manifests itself in sepsis, a weak novocaine solution of 0.5% is administered intravenously. Take 0.5 ml of the mixture per 1 kg of weight.
• A flour mash is also used, which is poured through a probe daily, 15–20 liters each.
• To strengthen the heart muscle, a mixture is prepared: 6 g of bromopotassium, 10 ml of valerian tincture and 15 ml of lily of the valley are added to 400 ml of distilled water. This is a single dose.

Prevention

FMD, like any viral disease, is easier to prevent than to treat.

The focus is on vaccination. Most often, a saponin composition is administered in an amount of 1 ml. It begins to act in 10-14 days, reaching a protective peak in a maximum of a month.

Immunity lasts from 6 months to a year. Cattle are revaccinated once a year, while pigs have to be vaccinated twice a year.

Age is also taken into account: in calves, for example, “maternal” immunity is very strong and the first 3 weeks can interrupt the effect of the vaccine. In goats and lambs, it is much weaker, and piglets are practically not protected.

The remaining preventive measures are largely traditional:

• regular cleaning of the premises with a change of bedding;
• placement of livestock according to the norms (no crowding);
• periodic examination of the animal, pay special attention to the oral cavity, the condition of the skin, coat and hoof plates;
• the use of high-quality feed, water and additives;
• limiting contact with already infected animals (for example, do not bring to the same pasture).

Is foot and mouth disease dangerous to humans?

As we remember, such a virus is reluctantly transferred to people, although its danger should not be underestimated.
The risk group includes people who work directly with animals: veterinarians, milkmaids, shepherds, workers in slaughterhouses and meat processing plants. But even in the courtyard you can get infected through contact or consumption of meat and milk from a sick "cattle".

Foot and mouth disease is a dangerous, acute, highly contagious viral disease of many animal species, characterized by fever, salivation, aphthous-erosive lesions of the mucous membrane of the tongue and oral cavity, skin of the nasal mirror, limbs, mammary glands, myocarditis and myositis with high mortality of young animals in the first days of life. FMD can also be transmitted to humans from animals.

History reference. For the first time foot and mouth disease was described and pointed out its contagiousness in Italy in 1546. D. Fracastoro. In the 17th and 18th centuries foot-and-mouth disease, which often proceeded malignantly and was often confused with rinderpest of those cattle, was established in many countries of Europe. FMD has been known in Russia since the first half of the 19th century. In 1897, Loeffler and Frosch discovered the causative agent of the disease, the filter virus.

Currently, foot and mouth disease is found in many countries in Europe, Asia, Africa and South America. The last outbreak of foot-and-mouth disease in Russia was recorded in 2005 in the Amur Region, Khabarovsk and Primorsky Territories as a result of its introduction through the territory of Mongolia from China, where there was an epizootic of foot-and-mouth disease of the Asia-1 type.

Economic damage from foot and mouth disease consists of losses as a result of the death of mainly young animals (calves, piglets, lambs), a decrease in the milk productivity of cows by 50-75%, a decrease in the live weight of sick animals and abortions. Especially huge losses are carried out by quarantine measures. So, type O foot-and-mouth disease in Taiwan in 1997, where there were more than 6 thousand foot-and-mouth disease foci and more than 4 million were destroyed. pigs, brought a total economic loss of about 10 billion US dollars. In the UK, a similar FMD epizootic in 2001, when 2030 FMD foci were registered and more than 4 million heads of animals were destroyed, the damage amounted to about 12 billion dollars. In Russia in 2005, due to FMD foci of the Asia-1 type in the Amur Region, Khabarovsk and Primorsky Territories, direct economic damage amounted to more than 45 million rubles.

The causative agent of the disease The virus belongs to the Picornaviridae family, genus Aphthovirus. The viral particle consists of ribonucleic acid (RNA) surrounded by a peripheral protein capsid of 32 capsomeres. In the body of a sick animal, the virus is in the highest concentration in the epithelium of the walls of aphthae and in the lymph in the first 24-48 hours of illness. It can, but in much lower concentrations, be found in saliva, blood, urine and feces. Moreover, the virus from aphtha becomes contagious already at a dilution of 1:100-200 million, and from feces only at a dilution of 1:100. 7 types of the virus are known: O, A, C, Asia-1, CAT-1, CAT-2, CAT-3 and many of their variants. Now in the world types O, A, Asia-1 and CAT-2 are predominantly distributed. Animals that have been ill with one type of foot-and-mouth disease may become ill again if they are infected with another type of virus.

FMD virus is relatively resistant to environmental factors. On the surface of objects contaminated with the secretions of animals with foot-and-mouth disease, the virus remains 150, in manure - up to 168, in slurry - up to 40, in sewage- up to 103 days. In blood cattle and pigs (after quick freezing of meat carcasses), the virus remains active for 40 days. In chilled milk it lasts up to 47 days, in fresh milk at a temperature of 37 degrees it dies after 12 hours, while in sour milk and when making cheese the virus quickly dies. The virus retains the coat of animals and clothing of people for 28-40 days. With properly conducted biothermal manure neutralization, the virus dies in 10-15 days. Heating up to 60 degrees kills the virus in 15 minutes, and at 80-100 degrees it is destroyed almost instantly. The best disinfectant to kill FMDV is to use 2-3% hot sodium hydroxide solution and 1% formaldehyde solution.

epidemiological data. Foot-and-mouth disease affects all types of artiodactyl animals. Cattle are most susceptible to foot and mouth disease, followed by decreasing pigs, sheep, goats and deer, buffaloes and camels are less sensitive. Cases of FMD infection of dogs and cats through the milk of cows with FMD are described. Birds and horses are not susceptible to FMD. Of the laboratory animals, guinea pigs, rabbits, and mice suffer from foot and mouth disease.

Animals of any age are ill with foot and mouth disease, however, young animals under the age of 2-3 months are more easily infected and more severely ill.

The source of the infectious agent is infected, sick animals, as well as animals - convalescents, which can be virus carriers for a long time.

The main ways of spreading infection. The FMD virus is transmitted mainly by alimentary and aerogenic routes. The FMD pathogen is introduced into the farms when sick or recovered animals are admitted to the farm by private household plots, peasant farms; in contact with sick (recovered) animals, including wild ones; when grazing, watering, hauling; with infected feed, water, as well as when using milk from sick animals; when importing products of slaughter of sick (recovered) animals (use of untreated kitchen waste in feed); spread with the wind (small particles of feed, affected tissue, saliva, dust, etc.), with infected (contaminated) care items; clothing and footwear for caregivers, vehicles.

Pathogenesis. The virus, which enters the body of an animal through the digestive tract or external integuments, penetrates through the mucous membranes into the epithelial cells, where it is fixed and multiplied. The reproduction of the virus causes a response of the body in the form of a serous inflammatory process. There is the formation of one or two primary aphthae, which are usually not noticed by the owners of the animals. The general condition of the animal in this phase of the disease usually does not change. After 24 hours, in most animals, the disease enters the second phase. From the places of primary localization, the virus, after reproduction, enters the bloodstream, and then to all organs and tissues. Generalization of the process causes an acute febrile reaction in the animal.

The protective agents available in the animal's body neutralize the virus in the blood in most organs and tissues, but due to the fact that the epidermis is relatively poorly supplied with blood, and hence with antibodies, the virus begins to multiply in it. As a result of intensive reproduction of the virus in the epidermis, multiple secondary aphthae appear in the oral cavity, in the region of the interhoof gap and the corolla of the limbs, on the skin of the udder teats (generalized process). FMDV can replicate in the heart and skeletal muscles.

The course and symptoms of the disease. All animals in vivo FMD is usually acute. In adult animals, an abortive course of foot and mouth disease is sometimes observed, accompanied by a short-term increase in body temperature and a rapidly onset recovery. In adult animals, FMD usually proceeds benignly. It is customary to distinguish between typical and atypical forms (malignant, abortive and latent) foot and mouth disease.

In cattle the incubation (hidden) period is 1-3 days, but can be from 12 hours to 7 days, reaching in some cases up to 14-21 days.

In a benign course, we first note a decrease in appetite, lethargy of chewing gum, increased salivation. Then, in a sick animal, an increase in body temperature to 40.5-41.5 degrees is observed, the pulse quickens, the state of the animal becomes depressed, there is a refusal to feed and the absence of chewing gum. By the second and third days of illness, on the inner surface of the upper and lower lips, on the toothless edge of the lower jaw, on the tongue and buccal mucosa, aphthae. Almost simultaneously, some animals form aphthae in the area of ​​the interhoof gap and on the skin of the udder. With foot and mouth disease, all four limbs are often affected, but there are cases when only two front or two rear limbs are affected.

At the beginning of the disease, aphthae are the size of a millet grain, then they merge and increase to the size of a pea or walnut. After 12-24 hours, the walls of the afts break, leaving behind fresh erosion. At this point, the body temperature of the animal drops to normal. When examining a sick animal, we note profuse salivation, a foamy mass and a characteristic smacking are formed in the corners of the mouth. The resulting erosions heal in 6-8 days, but in the case when the process is complicated by a secondary infection, the healing of erosions is delayed up to 2-3 weeks.

On the extremities, foot-and-mouth disease begins with painful and hot swellings appearing on the skin of the corolla and crumbs, in the region of the interhoof gap, due to which the animal develops lameness. If all limbs are affected, then such animals lie down and can be raised with great difficulty. In the future, aphthae appear at the site of the resulting swelling, which soon burst with the release of the contents in the exudate aphthae. With timely, properly started treatment and keeping animals on dry litter, erosion heals within 5-8 days. If aphthous lesions are extensive, then the animal develops phlegmon of the corolla, deep purulent pododermatitis, purulent arthritis, up to the subsidence of the horn shoe.

In lactating cows, the formation of aphthae of various sizes is often observed on the skin of the udder and teats. After opening the aphthae, erosion remains. The inflammatory process tends to spread to the top of the nipple and to the mucous membrane of the nipple canal. All these inflammatory processes lead to dysfunction of the affected quarter of the udder, which is manifested by a change in the composition of the milk, the milk becomes slimy, acquires an acidic reaction and a bitter taste. As a result of blockage of the teat canal with fibrinous, casein plugs and scabs, leading to difficulty in the exit of milk, mastitis develops in cows. In lactating cows, milk production is reduced to 75%. Milk productivity, with timely and correctly started treatment, is restored in cows slowly, sometimes it takes several months. In some adult animals, we note a disorder of function digestive tract accompanied by diarrhea.

In calves under 2 months of age, foot-and-mouth disease usually occurs in an aphthous form, but with symptoms of acute gastroenteritis. If the necessary therapeutic measures are not taken in a timely manner, the disease will end in the death of calves. In some calves, as a result of a complication of secondary microflora, foot and mouth disease is complicated by bronchopneumonia.

With a benign course, foot and mouth disease continues for 8-10 days. If complications join foot and mouth disease, then the disease stretches for 25 or more days.

Complications in animals with foot-and-mouth disease arise as a result of the attachment of pathogens of secondary infections to the foot-and-mouth disease process: streptococci, staphylococci, as a result of which purulent pododermatitis, corolla phlegmon, endometritis, mastitis, nephritis, bronchopneumonia, etc. appear in sick animals. Complications mainly develop in animals with weak body resistance. The FMD virus itself can lead to the development of disorders in the activity of the cardiovascular system due to myocardial dystrophy and metabolic disorders.

There are cases when foot and mouth disease can have a malignant course. In the malignant form of foot-and-mouth disease, when the disease initially proceeds with symptoms typical of foot-and-mouth disease, on the 8-12th day (in the recovery stage), the animal suddenly begins sharp deterioration the state of the animal. In the animal, we note weakness, depression, the pulse quickens to 120-140 beats per minute, the animal refuses to feed, chewing gum stops. In some animals we note paralysis of the hind limbs. Death in a malignant form of the disease occurs from cardiac arrest.

In some cases, foot and mouth disease acquires a malignant course from the very beginning of the disease, when the body temperature rises by 0.5-1 degrees, against the background of weak aphthous lesions with simultaneous damage to the udder. The disease proceeds with loss of appetite, severe depression and disorder of cardiac activity. Mortality in animals is 20-50%.

Pigs. In pigs, the incubation (hidden) period is most often 24-48 hours, but sometimes it is delayed up to 8 days. Among pigs, the disease is acute, with high mortality of young animals. The disease is characterized by fever, depression and decreased appetite. In pigs, mainly the limbs are affected, lameness appears, in some pigs we note the fall of the hooves. Aphthae appear on the patch, mammary glands, occasionally in the oral cavity. After the rupture of the aphthae, erosion remains. In adult pigs, the disease lasts 8-25 days. In piglets, foot-and-mouth disease occurs in a septic form, leading to the death of 60-100% of animals already in the first days of the disease. At severe course diseases occur hemorrhages in the mucous membranes of the digestive tract, lungs and kidneys, under the serous membranes.

Sheep the incubation (hidden) period lasts 2-3 days. The disease is less acute than in cattle. We rarely note such a symptom characteristic of foot-and-mouth disease as salivation. Aphthae small, open early and in the absence of complications quickly heal. In connection with the defeat of the limbs (aphthae in the area of ​​the interhoof gap and corolla, as well as pododermatitis), lameness occurs in sheep. With the mass distribution of foot-and-mouth disease in the flock, in some sheep we reveal a characteristic symptom complex: aphthous-erosive changes on the lips, tongue, gums, edentulous margin upper jaw, on the limbs and udder, increased body temperature (up to 41.5 degrees), decreased appetite, periodic cessation of chewing gum, depression. It often happens that foot and mouth disease in sheep in flocks occurs with mild signs or these signs are completely absent, as a result, foot and mouth disease remains undiagnosed. Sheep with such a development of foot and mouth infection can remain virus carriers for several months, acting as a hidden source of the virus. In lambs, foot and mouth disease often occurs in the form of septicemia and is accompanied by a large case.

Goats the incubation (hidden) period lasts from 2 to 8 days. It is less acute than in cattle. In the first days of the disease, we note fever, depression, loss of appetite, damage to the mouth and limbs, which leads to lameness. The mouth of sick goats is closed, we note the gnashing of teeth. Abundant salivation is absent. Often the udder is damaged. Recovery usually occurs within 10-14 days. Goats are more resistant to foot-and-mouth disease, but sometimes they also have a malignant course.

Deer with foot and mouth disease, we note diarrhea, damage to the mucous membranes of the oral cavity and extremities, which are often complicated by necrobacteriosis. In the absence of complications, deer recovery occurs in 10-12 days.

pathological changes. We find aphthae and erosion in the oral cavity, sometimes on the mucous membrane of the esophagus and in the proventriculus in ruminants. In piglets, lambs and calves, at autopsy, we find hemorrhagic inflammation of the intestine, degenerative changes in the heart muscle (tiger heart).

In the malignant form of the course of foot and mouth disease in the heart muscle, we find severe lesions. The cardiac muscle becomes pale and flabby, grayish-reddish spots and caseous degenerate gray-white foci of various sizes appear on the cut. Under the epicardium and endocardium we find hemorrhages. In the liver, kidneys and skeletal muscles - degenerative changes.

Diagnosis as with all infectious diseases is put complex taking into account epizootological data, clinical signs of the disease, pathoanatomical changes and mandatory laboratory test results.

For laboratory research send walls and contents of fresh aphthae (lymph), blood samples at the time of fever in animals, lymph nodes of the head area, esophago-pharyngeal mucus and blood serum samples (not earlier than 14 days after the onset of clinical signs). The selected materials are placed in closed sterile vials, frozen or transported in a preservative liquid (equal volumes of neutral glycerol and 0.8% sodium chloride solution) in thermal containers with ice (refrigerant) with strict observance of precautionary measures (safety of working with microorganisms of 1-2 pathogenicity groups (hazards). Sanitary and epidemiological rules SP 1.3. 1285-03).

Materials for research sent by courier with cover letter detailed description epizootic situation in the economy (settlement).

Laboratory diagnostics foot-and-mouth disease is based on the isolation and identification of foot-and-mouth disease virus from samples of pathological material (walls and contents of aphthae, lymph, lymph nodes, meat samples, meat products and feed), detection of the pathogen antigen and viral RNA, determination of the primary structure of the BP 1 gene of the foot and mouth disease virus and the detection of post-infection antibodies (studies in the RSK, ELISA, PCR, RMN, virus isolation).

differential diagnosis. When making a diagnosis of foot and mouth disease, veterinarians should exclude feed stomatitis and, vesicular stomatitis kr.r.sk.,. In sheep and goats, we exclude contagious pustular stomatitis and.

Treatment. Given that the success of treating animals with foot and mouth disease largely depends on strict adherence to the rules of feeding and keeping, we provide peace to sick animals in order not to overstrain the heart. Rooms for sick animals should be clean, with sufficient bedding material and have a constant supply of fresh air. In order to combat dehydration, sick animals should receive plenty of clean cool water in agricultural enterprises, household plots, peasant farms. The diet should contain soft digestible feed (grass, flour talker, good silage in winter, soft hay). The mouth is washed clean water with the addition of 2% acetic acid, a 0.1% solution of potassium permanganate, a 0.5% solution of furacilin can be used. In case of severe damage to the oral mucosa, we use an ointment (anesthesin 2.5 g; novocaine 2.5 g; blue vitriol 5g; fish oil 20g; vaseline 70g). This ointment accelerates the healing of erosions and, having an analgesic effect, allows animals to take food.

The limbs are cleaned of dirt and every 1-2 days the hooves, the whisk, the skin of the arch of the interhoof gap are lubricated with tar in half with fish oil. For the same purpose, animals are passed through disinfectant barriers with sawdust, which are impregnated with tar, or through baths with a 5% formalin solution.

In case of severe damage to the limbs (phlegmon of the crumb, corolla, interdigital fiber), the inflamed areas are smeared with tincture of iodine. We clean the hooves, remove dead tissues, ulcers and wounds, cauterize with potassium permanganate powder in half with streptocide and apply a protective bandage or use shoes made of tarpaulin and other dense material. In the event that foot and mouth disease in an animal is complicated by sepsis as a result of secondary infection, intravenously we introduce a 0.5% solution of novocaine at the rate of 0.5 ml per 1 kg of animal weight. Antibiotic therapy is used, incl. modern antibiotics cephalosporin series. To prevent udder aphthous lesions in cows, milkmaids must keep their hands clean and follow the rules for milking cows. With aphthous lesions on the udder, trypoflavin-novocaine ointment is used (trypoflavin 1g, novocaine 4g, vaseline 100g), synthomycin emulsion or 15% propolis ointment on vaseline.

In case of severe foot and mouth disease and cardiac disorders, the use of a mixture is recommended: valerian tincture 10 ml, lily of the valley tincture 15 ml, potassium bromide 6 g, distilled water 400 ml; inside at one time.

At malignant forms foot-and-mouth disease sick cows must be injected daily through a tube or with a bottle of 20-30 liters of flour mash. Weakened animals are not bad to give honey 100-200g or drink back with the addition of 200-400g of sugar.

From specific means, cited blood of convalescents or serum is used for animals at the rate of 2 ml per 1 kg of weight. These tools are effective if applied before the generalization of the process. For therapeutic purposes, you can also use anti-foot and mouth immunolactone.

Immunity and specific prophylaxis. In newly vaccinated animals, immunity is formed by 21 days. Virus-neutralizing antibodies in the blood serum appear 5-7 days after infection, reaching a maximum after 3-4 weeks and can persist for about a year. Collostral antibodies in calves persist for 3-5 months.

For prophylactic immunization of animals, depending on the epizootic situation, inactivated mono-, bi- and polyvalent vaccines are used according to certain schemes. A stable level of post-vaccination antibodies that protect adult animals from FMD is maintained for 6 months.

Control and prevention measures. FMD well-being in the Russian Federation has been achieved through the implementation of a program that includes monitoring the epizootic situation, controlling the movement of animals and products of animal origin, taking measures to prevent the introduction of FMD virus to livestock farms, observing the “closed type enterprises” regime, conducting preventive vaccination animals in areas of high risk of introduction and spread of the disease, the elimination of sick or all animals in the foci of infection, strict adherence to the system of quarantine measures. In the event of an outbreak of foot-and-mouth disease, by the decision of the Governor of the region on the farm, settlement quarantine is imposed. Quarantine and restrictive measures are being taken in accordance with the instructions "On measures to prevent and eliminate the disease of animals with foot-and-mouth disease", approved by the Main Directorate of Veterinary Medicine of the USSR Ministry of Agriculture dated March 15, 1985.

Preventive vaccination of animals is carried out in threatened areas with subsequent control of the immune background. Specific prophylaxis of foot-and-mouth disease is carried out using inactivated vaccines produced by FGU "ARRIAH" and FGPU "Shchelkovsky Biokombinat".

In the Vladimir region, for the purpose of all kinds of emergencies for foot-and-mouth disease, a 30-kilometer buffer zone has been created around FGU VNIIZhZ, which includes Sobinsky, Suzdalsky districts and the city of Vladimir, where vaccination of the entire livestock of the kr.r.sk. and small cattle incl. and those in the private sector. The veterinarians of these regions constantly monitor the intensity of immunity by conducting monitoring studies of blood samples at the FGU "ARRIAH".

Ministry of Agriculture and Food of the Republic of Belarus

Educational Establishment "Vitebsk Order" Badge of Honor"

State Academy of Veterinary Medicine

Department of Epizootology and Infectious Animal Diseases

foot-and-mouth disease farm animal infectious

On the topic: FMD

VITEBSK 2010

Plan

1. Definition of disease.

History reference.

The spread of the disease.

Economic damage.

Etiology.

epidemiological data.

Pathogenesis.

Course and symptoms.

pathological changes.

Diagnostics.

Differential diagnosis.

Treatment.

specific prophylaxis.

Measures for the prevention and elimination of the disease.

foot and mouth disease

(lat. - Aphtae epizooticae; English -Foot-and-Mouthdisease)

Definition of disease.

FMD - highly contagious, acute infectious disease artiodactyl farm and wild animals, as well as callosities (camels), characterized by fever, the development of aphthous lesions on the mucous membrane of the oral cavity, the skin of the extremities in the corolla and crumbs, less often on the udder. AT exceptional cases foot-and-mouth disease affects other animal species, except artiodactyls. If personal prevention measures are not followed, people get sick with foot and mouth disease, children are most susceptible.

History reference.

It is officially believed that the first mention of foot and mouth disease in the literature was made by the Italian scientist D. Fracastoro in 1546. He described a disease that affected mainly cattle and was accompanied by signs characteristic of foot and mouth disease. German researchers D. Heffler and P. Frosch (1897) found that the causative agent of foot and mouth disease is a virus. With this, they opened a new era in microbiology and laid the foundation for veterinary and medical virology. The multiplicity of types of foot-and-mouth disease virus was established by French scientists A. Carré and A. Balle in 1922. Attempts specific prevention foot-and-mouth disease in animals was described in 1781. In Prussia, according to the official decision of the Higher Sanitary Board, vaccination against foot-and-mouth disease was carried out in cattle. Its principle was to infect healthy animals with foot-and-mouth disease, in which a cotton thread moistened with the saliva of sick animals was injected under the skin in the back area. In Russia, in accordance with the instructions for vaccinating cattle against foot-and-mouth disease (1909), it was recommended to pull through auricle thread moistened with the saliva of sick animals. The first vaccine for the specific prevention of foot-and-mouth disease in animals was obtained in 1934 by F.I. Hansen and W. Jensen. FMD epizootics and panzootics have been repeatedly noted in many countries of the world, including the territory of the USSR. All this dictated the need to develop a scientifically based system of measures to combat foot-and-mouth disease, to concentrate efforts veterinary service, scientists and practitioners on the elimination and prevention of such a dangerous disease. For this purpose, in 1958, the All-Russian Scientific Research Institute of Foot and Mouth Disease of the USSR Ministry of Agriculture was established, now the Federal State Institution "Federal Center for Animal Health" (FGU "ARRIAH"). In 1995, the OIE FGU "ARRIAH" was awarded international status"OIE Regional Reference Laboratory for FMD for Eastern Europe, Central Asia and Transcaucasia". The specified laboratory carries out scientific support of measures for the prevention of foot-and-mouth disease in the Republic of Belarus.

Spreading.

According to the OIE, annually in the world 10-80 countries are unfavorable for FMD. Information about the incidence of foot-and-mouth disease in the Republic of Belarus dates back to the middle of the 19th century. Since 1983, the republic has been free of foot and mouth disease.

Economic damage.

Such damage from foot-and-mouth disease amounts to billions of rubles due to its rapid and widespread spread, a decrease in the productivity of adult animals, the death of young animals, and the high costs of quarantine measures. Foot and mouth disease is a biological catastrophe, the economic damage is ten times greater than the damage from natural disasters such as earthquakes, floods, hurricanes, etc. For example, foot-and-mouth disease in Taiwan in 1997, where more than 6,000 foot-and-mouth disease foci arose and over 4 million pigs were destroyed, caused a total economic loss of about 10 billion US dollars. An epizootic of foot-and-mouth disease type O in the UK in 2001 caused an economic loss of about $12 billion.

Etiology.

FMDV is an RNA-containing virus that belongs to the family Picornaviridae, genus Aphtovirus. Currently, 7 serological types of the virus are known (A, O, C, CAT-1, CAT-2, CAT-3 and Asia-1). Each type has a certain number of subtypes (options): type A has 46 options, O - 14, C - 5, CAT-1 - 9, CAT-2 - 3, CAT-3 - 4, Asia-1 - 3 options. Types O, A, Asia-1 and CAT-2 are predominantly distributed. The antigenic variability of the pathogen or, as it is also called, illuriality (from Latin pluralis - plurality) in foot and mouth disease is of great practical importance, since infection with one type or subtype of the virus and the disease does not create immunity against another type of virus or even its subtype. In this case, the transition of one type of FMD virus to another is possible. In this regard, the production of FMD vaccines that provide immune protection against most types and subtypes of FMDV remains problematic. FMDV can be maintained by serial passages in naturally susceptible animals or laboratory animals (guinea pigs, white suckling mice, rabbits and tissue culture). The most sensitive to the virus is the cell culture of the primary trypsinized kidney of a cow, sheep, pig, and the transplanted line VNK-21, on which CPE manifests itself after 6 hours and reaches a maximum by 18-24 hours after infection. The virus also multiplies well in the culture of kidney cells of susceptible animals, in the culture of explants of the epithelium of the tongue and the rumen of cattle, and in some transplanted cell lines BHK-21, JBRS-2, JFFA-3, SPEF, etc. FMDV is relatively resistant to physical and chemical factors. High temperatures have a detrimental effect on the virus: at + 100 ° C, the virus dies instantly; at + 64°С - for 3 s; at +49°С - for 1 hour; at +37°С - in 21 hours. Low temperatures preserve the virus: in frozen meat, the virus can persist for up to 200 days; at -70°C it persists for several years. long time the pathogen persists in meat and dairy products: in chicken legs - for 112-119 days; in shoulder fat - 155-169 days; in bone marrow- 169-179 days; in ham fat - 176-183; in bacon - 183-190; in chilled milk - 14-40 days. On the coat of animals and human clothing, the virus persists for 40-145 days. It is stored in slurry for up to 30 days, and in sewage - up to 103 days. In a haystack, the virus persists for up to 6 months, in bran for up to 140 days, and in straw for up to 3 months. A good disinfectant effect in foot and mouth disease is provided by 2-3% solutions of sodium and potassium hydroxide, as well as formalin.

epidemiological data.

About 100 species of artiodactyls are susceptible to the FMD virus. Cattle are the most susceptible with almost 100% incidence. High susceptibility to foot-and-mouth disease virus has also been established in pigs, then in sheep and goats. Deer, buffaloes, camels and yaks are less susceptible. Of the wild artiodactyls, moose, saigas, roe deer, wild boars, giraffes, bison, llamas, etc., suffer from foot and mouth disease, and they have a very important role in the spread of foot-and-mouth disease. Other types of animals (dogs, cats, rats) get sick only rarely. In general, horses and birds are not susceptible. Susceptible animals of all age groups However, young animals, obese and well-fed animals are more susceptible. Under laboratory conditions, guinea pigs, mice, and rabbits can be infected with the FMD virus. A person (children) is susceptible to FMD. The source of the causative agent of infection is animals with foot and mouth disease and virus carriers. Virus carrying in cattle can last from 240 to 400 days. Long-term carriage in sheep - up to 330 days. Vaccination does not stop the virus carrier. The virus is released into the external environment with all excretions and secrets (saliva, milk, feces, urine). Saliva is especially rich in virus. Important from an epizootological point of view is the fact that the virus is excreted from the body of animals during the incubation period of the disease. Transmission factors are virus-contaminated feed residues, water, bedding, manure, care items, transport, etc. Unsusceptible animals - dogs, cats, horses, poultry and wild birds, etc. can carry the pathogen over a considerable distance. In the distribution of foot-and-mouth disease, products and raw materials of animal origin play a special role: milk, meat, leather, wool, etc., contaminated with foot-and-mouth disease virus. The so-called "milk epizootics" are described, which arose as a result of feeding animals with non-decontaminated milk and skim milk. For tens and even hundreds of kilometers, the FMD virus can spread with air masses and infect animals in an aerogenic way. The gates of infection are the mucous membrane of the oral cavity, the skin in the area of ​​the corolla, interhoof gap, udder (hairless part of the skin). Easier infection occurs when their integrity is violated. The disease is seasonal. FMD most often occurs in autumn-winter period. This is due to the greater preservation of the virus in the cold season and the period of the most intensive sale, purchase of animals, their delivery to a meat processing plant, etc. FMD is characterized in some cases by stationarity, which is most often due to the poor effectiveness of measures taken to eliminate the disease and the carriage of the virus by recovered animals (usually sheep). With foot and mouth disease, there is a 5-7-year periodicity, which is based on the variability of the virus and an increase in its virulence, provided that specific prophylaxis has not been carried out in the area. Long-term carriage of the virus in the body of animals (recovered and vaccinated), its long-term preservation in the external environment, a wide range of susceptible domestic and wild animals (up to 100 species), the multiplicity of types and subtypes of the virus that do not create cross-protection - all these factors ensure stability pathogen, its preservation in nature and reproduction of the epizootic process. FMD occurs in the form of epizootics or panzootics. Lethality in cattle is 0.2-0.5%, with malignant course- up to 50%, sometimes - up to 100%.

Pathogenesis.

The virus enters the body of an animal in different ways (aerogenic, alimentary, with direct contact). As predominantly dermatropic, he finds himself optimal conditions for reproduction in the middle layers of the epithelium of the mucous membranes or on hairless areas of the epidermis. In these places, the foot-and-mouth disease virus multiplies, causing the formation of so-called primary aphthae. There are usually one or two of them. The content of the aft contains the pathogen in high concentration, its infectious titer can reach 10-10. From the primary aphthae, the virus enters the bloodstream via lymphogenous pathways, which facilitates the transport of the virus to other organs and tissues. 48 hours after infection, against the background of general fever, on the so-called favorite places - hairless areas of the skin (nasal mirror, nostrils, udder), in the oral cavity, esophagus, scar and on the skin in the hoof area, and in pigs on the snout and limbs, secondary aphthae. In malignant foot-and-mouth disease, the virus penetrates the skeletal muscles and heart, multiplies there, causing functional disorders of the heart and tissue defects. Sometimes the CNS is also involved in the pathological process. The stage of generalization lasts 2-4 days. Starting from the 4th day, the blood again becomes free of the virus. Body temperature drops, which is associated with the appearance of antibodies. The shells of primary and secondary aphthae rupture several hours after their appearance, and ulcers form in their place, and then erosion. The formation of aphthae on the udder contributes to the occurrence of mastitis, on the skin of the corolla - lameness. If the resistance of the organism is high, then the process can stop at the stage of primary aphthae, which is extremely rare. With low resistance of the organism, the virus does not linger at the site of primary penetration, primary aphthae are not formed, and the disease is septic in nature. This is more common in young animals, whose mortality rate is high.

Course and symptoms.

The incubation period lasts for foot and mouth disease from 2 to 7, and sometimes up to 14-21 days. Infected animals during this period already release the virus into the environment and pose a danger to other animals. In cattle, there is a benign and malignant course of foot and mouth disease. In a benign course, the primary symptom of the disease is a decrease in appetite. Then fever appears, the body temperature rises to 40.5-41.5 ° C. Animals are oppressed, refuse to feed, pulse and respiration are quickened, milk yield is sharply reduced. In the initial period of the disease, the mucous membrane of the mouth is dry, hot, its hyperemia (redness) is observed. On the 2-3rd day after the rise in body temperature in the oral cavity, aphthae (vesicles) appear on the tongue, on the wings of the nose, and sometimes on the nasal mirror, filled at first with a clear, and then with a cloudy liquid. Then the walls of the bubble rupture after 1-3 days, the lymph contained in them mixes with saliva and is released outside. In place of bursting bubbles are formed painful erosions with uneven edges, which heal in 5-8 days. Body temperature with the appearance of aft quickly decreases. During the period of fever and the appearance of secondary aphthae, animals secrete copious amounts of saliva. The saliva is viscous, stretches to the floor, the animals smack their lips in a peculiar way. Aphthae are also formed on the skin of the extremities in the region of the interhoof gap and corolla, which is accompanied by lameness. In the presence of good bedding, healing of the skin on the extremities occurs after 7-12 days, and with poor care and maintenance, the process is complicated and may be accompanied by suppuration and detachment of the horny shoe. In cows, the infectious process may be accompanied, in addition to the above, by damage to the udder. The skin on the nipples turns red, swells, small aphthae appear, which then merge, reaching a size hazelnut. The aphthae formed on the nipples burst during milking and painful erosions remain in their place, the process can be complicated by purulent mastitis. The milk of sick cows becomes slimy, bitter in taste, coagulates easily and hardly churns into butter. The decrease in milk yield in animals with foot and mouth disease can reach 50-75% in the herd. Due to damage to the oral cavity, udder And lower extremities food intake is disturbed, diarrhea, lameness are sometimes observed, body weight of animals decreases. Recovery usually occurs within 3-4 weeks. Similar symptoms with foot and mouth disease are noted in sheep and goats, however, the course of the disease in this species of animals is more benign. In pigs, foot and mouth disease affects the limbs and snout, and in suckling sows, the udder. The disease of the extremities is accompanied by lameness and often the fall of the hooves. The death of suckling pigs from foot and mouth disease can reach 60-80%. Sometimes in cattle and pigs there is a severe malignant course of foot and mouth disease. It is accompanied by severe weakness, depression, trembling, increased respiration and pulse. Sometimes recovered animals suddenly experience a sharp deterioration in their condition and sudden death due to heart failure. The death of animals can reach 50-70%, and sometimes 100%. FMD can affect people, especially children. Infection of people most often occurs when the virus enters with food (milk). After 3-4 days incubation period a person has red spots, then bubbles filled with a cloudy liquid. The blisters burst and ulcerations form in their place. Bubbles can also appear on the flexor surfaces of the fingers, on the lower leg and forearm. In adults, the course of the disease is benign, while in children it is severe. They develop diarrhea, aphthous lesions in the oral cavity are not always observed.

pathological changes.

Specific signs of foot and mouth disease in cattle are aphthous lesions of the mucous membrane of the oral cavity, less often the skin of the interhoof gap, corolla, mammary gland, nasal mirror. Aphthae can be found on the mucous membrane of the scar, less often on the anus and vagina, and as an exception on the skin of the trunk. In pigs, aphthae are rarely found in the oral cavity, more often they are recorded on the skin of the patch, corolla, and interhoof gap. In small ruminants, aphthae in the oral cavity can be small. Quite often in sheep, the skin of the corolla, interhoof gap and crumbs are affected. Aphthae can be from a pinhead to the size of a chicken egg. Of the other organs in foot and mouth disease, the abomasum and intestines are more often affected. They find catarrhal inflammation, often taking on a hemorrhagic character. In the abomasum, in addition, there may be dark brown scabs and ulcers. In some cases, hemorrhages are found on the epi - and endocardium, peritoneum, mucous membrane of the abomasum and small intestine, catarrhal mastitis. In a malignant course, aphthous lesions are poorly expressed. The main changes are found in the heart muscle and skeletal muscles. Active muscles are more often affected: hip, shoulder, intercostal, muscles of the tongue, etc. In the heart, during external examination, multiple grayish or grayish-yellowish foci of various sizes and shapes are noted. Their presence gives the heart a peculiar striped color, reminiscent of the skin of a tiger (tiger heart).

Diagnostics.

It is based on taking into account the characteristics of the disease (almost 100% incidence of animals, rapid spread, susceptibility mainly to artiodactyls, lack of seasonality, etc.). This disease is characterized by clinical signs. The final diagnosis of the disease is based on the results of laboratory tests. When conducting them, it is imperative to determine the type and variant of the FMD virus that caused the disease. This is important for the selection of appropriate vaccines for active specific prophylaxis. The walls and contents of afts (lymph) on the mucous membrane of the tongue (cattle), on the patch (pigs), on the skin of the corolla and interhoof gap (cattle and small cattle, pigs, camels and other species susceptible to foot and mouth disease) are sent to the laboratory for research. animals). In the absence of aphthae, blood samples are taken to isolate the virus at the time of the temperature reaction in animals, as well as the lymph nodes of the head and pharyngeal ring, the pancreas and the muscle of the heart (corpses of young animals of all animal species). Aphthae and lymph are taken in an amount of at least 5 g. The amount of other materials intended for virus isolation and its subsequent identification must be at least 10 g. Laboratory diagnostics involves the isolation of foot-and-mouth disease virus from samples of pathological material using cell cultures and sensitive laboratory animals. Identification of the virus is carried out in the RSK, ELISA and PCR. Detection of FMD virus antigen in suspensions of aphthous material, meat products, cell cultures is carried out using ELISA. The determination of the level of specific antibodies in the blood serum of recovered animals is carried out using pH (micromethod) and ELISA, and the identification of post-infection antibodies is carried out in the ELISA reaction. The diagnosis of foot-and-mouth disease is considered definitively established on the basis of laboratory tests in one of the following cases: isolation of the virus and its identification; detection of the corresponding antigen; detection and identification of post-infection antibodies. Diagnosis must necessarily include determining the type of FMD virus and its subtype that caused the disease in animals.

Differential diagnosis.

FMD in cattle should be differentiated from vesicular stomatitis, plague, malignant catarrhal fever, infectious rhinotracheitis, viral diarrhea, smallpox, necrobacteriosis, papular stomatitis and bluetongue, in pigs - from vesicular stomatitis, vesicular disease and vesicular exanthema, in sheep - from vesicular stomatitis, plague, infectious catarrhal fever.

Treatment.

Except in countries where FMD is highly prevalent, sick animals are not treated. The veterinary and sanitary rules for the prevention and elimination of foot-and-mouth disease, in force in the Republic of Belarus, provide for the destruction of animals sick with this disease.

specific prophylaxis.

It has a number of features: immunization must be carried out with a vaccine containing a strictly appropriate type and subtype of foot-and-mouth disease virus isolated in a particular farm; vaccination does not stop the carrier of the virus in animals, and they may pose a danger in terms of the spread of foot-and-mouth disease virus; vaccines may have residual reactogenicity and cause complications in the form of foot-and-mouth disease in animals; vaccination does not provide 100% protection of animals against FMD; in accordance with the OIE requirements, a country can be declared free from FMD, provided that in this country the vaccination of animals against this disease is not carried out. In the Republic of Belarus, vaccination of animals against foot-and-mouth disease has not been carried out since 1987. To provide Russia, and, if necessary, the CIS countries, including the Republic of Belarus, FGU "ARRIAH" produces the following vaccines: sorbed (GOA - saponin) vaccines for immunization of large cattle yaks, buffaloes, camels, sheep and goats: monovalent (O, A, C, Asia-1, CAT-1, CAT-2, CAT-3); bi-, three - and tetravalent (O, A, C, Asia-1); heptavalent and polystrain various combinations; emulsion vaccines for immunization of pigs: monovalent (O, A, C, Asia-1); bivalent (O, A; O, Asia-1; A, Asia-1); trivalent (O, A, Asia-1); emulsion monovalent vaccines for immunization of pigs and cattle based on multiple emulsion (O, A, C, Asia-1); universal concentrated mono - and polyvalent vaccines for early protection of animals, with the formation of immunity in the first four days after vaccination (immunity lasting one year or more).

Measures for the prevention and elimination of foot-and-mouth disease.

They are based on preventing the FMD virus from entering farms or states that are safe for this disease. The main reasons for the spread of foot-and-mouth disease in present stage are: the introduction of the virus from certain countries that are unfavorable for this disease, primarily in connection with the illegal importation of animals, livestock products and feed; human migration (tourism, pilgrimages, natural disasters, military conflicts, etc.), wild animals and birds; increased traffic of vehicles, including trucks, etc. In this regard, priority measures for the prevention of foot-and-mouth disease should include the implementation of an appropriate state program that provides for monitoring the epizootic situation and its prediction, control over the movement of animals and products of animal origin, the implementation of measures to prevent the introduction of foot-and-mouth disease virus into livestock farms, compliance with the regime " closed type enterprise”, the annual conduct of exercises for rapid response in the event of foot-and-mouth disease and other activities that ensure the well-being of individual farms and the state as a whole. Currently, in various countries, depending on the epizootic situation, geographical conditions, livestock breeding methods, development level and other factors, three main systems of measures for the prevention and elimination of foot-and-mouth disease are used:

) prophylactic immunization is not carried out, and in the event of foot-and-mouth disease - the slaughter of all susceptible animals in the outbreak without vaccination (stamping out - "pure method of control");

) prophylactic immunization is not carried out, and in the event of foot-and-mouth disease, animals are slaughtered with ring vaccination (vaccination of animals located in the areas around the epizootic focus);

) systematic vaccination of animals in certain areas, and in the event of foot and mouth disease, the slaughter of patients and ring vaccination. The second system of measures is taken as the basis for the prevention and elimination of foot-and-mouth disease in the republic. When foot-and-mouth disease occurs, the farm, farm, department, complex is declared unfavorable and quarantine is introduced. Under the terms of the quarantine, it is prohibited: import (input) and export (output) of animals from a quarantined farm; regrouping of livestock, procurement and export of products of animal origin; entrance to the farm by unauthorized persons; holding exhibitions, fairs, trade; export of milk and dairy products in a non-decontaminated form; passage of all types of transport through a disadvantaged point; departure of transport outside the quarantined zone. When organizing events, an epizootic focus, a disadvantaged point and a threatened zone are distinguished. An epizootic focus is a room or a farm, a separate courtyard, a summer camp where there are sick animals or livestock products obtained from animals sick or ill with foot-and-mouth disease are stored. In the epizootic focus, sick animals are slaughtered and destroyed, clinically healthy animals are vaccinated. Other measures in the foot-and-mouth disease focus include: fencing the territory of the focus with a fence or ditch, organizing one entrance and a round-the-clock post; the allocation of separate attendants to care for sick animals, providing them with overalls; equipment of rooms for disinfection of milk; daily disinfection of the territory and premises in which sick animals and care items are kept; deratization, scaring away birds, stray dogs, cats; burning corpses or burying them in trenches on the territory of the hearth; manure, food residues, bedding are subjected to biothermal disinfection or incineration on the territory of the outbreak. An unfavorable point for foot and mouth disease is considered to be a settlement according to the administrative division, where there is an epizootic focus. Threatened zone - these are settlements bordering on a disadvantaged point. The main activities in the disadvantaged point and the threatened zone are: vaccination of all susceptible animals against foot-and-mouth disease; strict veterinary and sanitary supervision over the procurement and export of livestock, feed raw materials; observance of the veterinary and sanitary access regime; carrying out educational work, etc. Quarantine is removed 21 days after the last case of a case or forced slaughter an animal with foot-and-mouth disease and final disinfection. Animals that have been ill with foot and mouth disease can be slaughtered only at a meat processing plant after 3 months, and not sick, but vaccinated - 21 days after vaccination.

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The invention relates to the field of veterinary virology and biotechnology and relates to a vaccine against foot-and-mouth disease type A. The vaccine contains avirulent and purified antigenic material from the Aphtae epizooticae virus strain, fam. Picornaviridae, genus Aphtovirus, serotype A, deposited in the collection of the Federal State Institution "VGNKI" under registration number No. 2045/Kyrgyzstan/2007-DEPA No. 146S and 75S immunogenic components of foot-and-mouth disease virus, adjuvants aluminum hydroxide with saponin and maintenance medium in effective ratios. The vaccine has high immunogenicity and is able to provide effective protection against the pathogen of foot-and-mouth disease circulating in the countries of the Transcaucasus, Central Asia, the Middle and Far East. 11 w.p. f-ly, 14 tab., 1 silt, 4 pr.

Drawings to the RF patent 2526570

The invention relates to the field of veterinary virology and biotechnology and can be used in the development and manufacture of an inactivated adsorbed vaccine against FMD type A.

FMD is an acute contagious viral disease artiodactyl animals. It is characterized by a tendency to wide distribution and epizootic course. The disease is accompanied by large losses of milk, meat and other types of livestock products, complicates commercial operations and economic activities. To ensure the sustainable well-being of the country in terms of FMD in Russian Federation a system of measures is being implemented, in which the priority is to prevent the introduction of foot-and-mouth disease virus into the territory, and in high-risk areas - vaccination. Currently, in the Russian Federation and the CIS countries, as a rule, inactivated sorbed vaccines are used for immunization of cattle and small ruminants against foot-and-mouth disease, and inactivated emulsion vaccines are used for immunization of pigs.

The technology for manufacturing an FMD vaccine from an inactivated virus begins with the selection of production strains based on an epidemiological analysis of the dynamics of FMD in the country and neighboring countries. When creating preparations for specific prophylaxis, appropriate types of foot and mouth disease virus are used and strains with a wide antigenic spectrum within a type with pronounced cross immunogenicity are selected. A strain with a wide range of immunogenicity and satisfying the requirements of the region is selected by testing it in a cross-protection reaction or, more commonly, in a cross-neutralization reaction. As a rule, a virus population is used as a production strain, which, together with the system and conditions of industrial cultivation, ensures a guaranteed and high accumulation of the 146S and 75S components of the virus and the production of an immunogenic vaccine.

In addition, the requirements for the stability of the virus in the process of purification from tissue components and concentration, as well as the preservation of the virus during inactivation and long-term storage, are imposed on the production strain.

The FMD causative agent has a significant antigenic variability of strains within the same serotype, which is detected at different time intervals and in different territories and depends on the species composition of the susceptible livestock, its immune status and many other various factors. The antigenic variability of FMDV is due to amino acid substitutions in polypeptide fragments (antigenic epitopes) exposed on the surface of capsid proteins. Shifts in the antigenic spectrum, corresponding to the renewal of the structure of a new field strain, can vary from insignificant, captured by monoclonal antibodies, to significant, recorded using traditional polyclonal immunoglobulins. Significant changes in the antigenic characteristics of the natural strain with highly likely cause weakening specific immunity induced by a non-homologous antigen. They also cause difficulties in strain-specific diagnosis.

As a result, there is a need to create new diagnostic tools and specific immunoprophylaxis.

Known strains of foot-and-mouth disease type A, used as production in the USSR and the Russian Federation over the past 50 years.

These include the following strains: A 7 No. 103, isolated in 1962 in the Kuibyshev region; A 7 No. 2, allocated in 1965 in the Tajik SSR; A No. 717/73, allocated in 1973 in the Stavropol Territory.

These strains were used to obtain diagnosticums and FMD vaccines used in various regions of the country.

However, after the eradication of foot-and-mouth disease caused by antigenically close virus strains, they were discontinued and are currently supported only in the collection of microorganism strains of the Federal State Budgetary Institution “ARRIAH”.

An inactivated sorbed vaccine against FMD type A is known, containing the active substance in the form of an avirulent and purified antigenic material from a strain A 22 No. 550 homologous to the infectious agent, obtained in a sensitive biological system, and target additives in the form of an adjuvant and a supporting medium in an effective ratio.

The strain A 22 No. 550 of foot and mouth disease virus was isolated in 1964 in Azerbaijan and is used in the Russian Federation as a production tool in the manufacture of specific prophylaxis and diagnostics used throughout Russia and in the post-Soviet countries.

Known inactivated adsorbed vaccine from strain A (Georgia) 1999/ No. 1721 FMD virus Aphtae epizooticae type A, isolated in 1999 in the Republic of Georgia.

Known inactivated adsorbed vaccine from strain No. 1707 "Armenia-98" FMD virus Aphtae epizooticae type A.

Vaccine against foot-and-mouth disease virus type A from production strain A No. 1707 "Armenia-98", used in the buffer zone of Transcaucasia.

The disadvantages of vaccines made from these strains are insufficient antigenic and immunogenic activity against epizootic FMDV type A isolates currently circulating in the countries of the Caucasus, Central Asia and the Middle East (Turkey, Iran, etc.) due to the existing significant antigenic differences.

The closest to the proposed invention in terms of essential features is an inactivated sorbed vaccine against FMD type A, containing the active substance in the form of avirulent and purified antigenic material from strain A (Georgia) 1999/ No. 1721, obtained in a sensitive biological system, and target additives in the form of adjuvants GOA with saponin and supporting medium in the ratio, mcg:

Strain A (Georgia) 1999 / No. 1721-DEP was isolated in 1999 in the Republic of Georgia and is used in the Russian Federation as a production one in the manufacture of specific prophylaxis and diagnostics used throughout Russia and in the CIS countries.

A suspension culture of VNK-21 cells is used as a sensitive biological system, and Earl's solution without serum with the addition of FGMS, GBCS and antibiotics at pH 7.4÷7.6 is used as a supporting medium.

Polyhexamethyleneguanidine (PHMG) is used to purify a virus-containing suspension from ballast impurities, and aminoethylethyleneimine (AEEI) is used to inactivate the virus. Sodium thiosulfate is added to the suspension to neutralize AEEI.

Avirulent and purified antigenic material from strain A (Georgia) 1999/ No. 1721 is a suspension predominantly of 146S and 75S immunogenic components of FMDV. GOA with saponin is used as adjuvants.

The main disadvantage of known vaccines, including the prototype vaccine, is antigenic differences from FMDV type A isolates isolated at different time intervals, and vaccines prepared on their basis provide effective protection of animals only against infection with a homologous virus.

In 2003, a new isolate of FMDV type A was isolated in Iran, significantly different from previously studied strains of this type. During 2005÷2006 FMD type A has become widespread in Western Asian countries - Iran, Turkey, Saudi Arabia and Pakistan. In February 2006, FMD type A of line Iran/05 entered Thrace, the European part of Turkey, which borders Bulgaria and Greece. Carrying out 100% vaccination of all ruminants with the A 22 strain in Thrace in February-March 2006 prevented the spread of FMD to neighboring Greece and Bulgaria, but after 3-4 months fresh cases of the disease appeared.

In this regard, it became necessary to obtain a new vaccine against foot-and-mouth disease virus serotype A from strain A No. 2045/Kyrgyzstan/2007 to ensure the safety of the territory of Russia and neighboring states from this pathogen.

The task of creating the present invention included the development of an inactivated sorbed FMD vaccine that creates effective protection for susceptible animals against the epizootic FMDV type A circulating in the countries of the Transcaucasus, Central Asia, the Middle and Far East.

The technical result from the use of the present invention is to expand the arsenal of inactivated adsorbed vaccines against foot-and-mouth disease type A.

The specified technical result is achieved by the creation of an inactivated sorbed vaccine against FMD type A, characterized by the following set of features.

The proposed vaccine contains in 1 cm 3 of the preparation: the active substance in the form of avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007-DEP, obtained preferably in a suspension culture of VNK-21 cells in an amount of at least 3.0 μg and target additives : GOA preferably in the amount of 11000.0÷15000.0 µg, saponin preferably in the amount of 500.0÷1500.0 µg and maintenance medium in the amount of up to 1000000.0 µg.

The original virus for obtaining strain A No. 2045/Kyrgyzstan/2007-DEP was isolated in 2007 in the Republic of Kyrgyzstan. The strain was obtained by multiple successive passages on sensitive hetero- and homologous cell cultures. The strain is adapted to primary trypsinized porcine kidney cells and continuous cell cultures of VNK-21, IBRS-2 and PSGK-30.

For the manufacture of a vaccine, a suspension culture of VNK-21 cells is preferably used as a sensitive biological system, and Earl's solution without serum with the addition of FGMS, GBCS and antibiotics at pH 7.4÷7.8 is used as a supporting medium.

To inactivate the virus, AEEI is used, which is added to the virus-containing suspension to a concentration of 0.025÷0.05%. Upon completion of inactivation, AEEI is neutralized by adding sodium thiosulfate to the suspension.

The resulting antigen is purified from ballast impurities using PHMG, which is added to the suspension to a concentration of 0.005÷0.007%.

Avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007-DEP is a suspension containing predominantly 146S and 75S immunogenic components of FMDV.

Quantitative and qualitative content of viral material is determined by turbidimetry.

To prepare the vaccine, a viral material containing at least 0.5 μg of 146S and 75S of the FMDV immunogenic components is used in 1 cm 3 .

The required concentration of 146S and 75S immunogenic components of FMDV in the proposed vaccine, which is at least 3 μg per 1 cm 3 of the finished product, is obtained by adding the calculated amount of adjuvant-sorbent GOA to the avirulent and purified antigenic material. The optimal content of GOA in 1 cm 3 of the finished product is in the range from 11000.0 µg to 15000.0 µg.

An additional 10% aqueous solution of saponin is added to the resulting concentrate at the rate of 1500.0 μg per inoculation dose. For example, for the optimal inoculation dose of 2 cm 3 the final concentration is 0.075%, which corresponds to 750.0 μg of its content in 1 cm 3 of the finished product.

The resulting vaccine is a light yellow liquid with loose sorbent sediment, which forms at the bottom of the vial during storage and easily breaks into a homogeneous suspension when shaken.

The present invention includes the following set of essential features that provide a technical result in all cases for which legal protection is sought.

2. The active substance in the form of avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007-DEP in an effective amount.

3. Target additives.

The features of the invention that characterize the proposed vaccine and coincide with the features of the prototype, including a generic concept that reflects the purpose, are:

1. FMD vaccine type A inactivated sorbed.

2. Active substance.

3. Target additives.

Compared with the prototype vaccine, the essential distinguishing feature of the proposed vaccine is that, as active substance it contains avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007 of FMDV type A in an effective amount.

The present invention is also characterized by other distinctive features expressing specific forms of implementation or special conditions its use.

1. Avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007, obtained in a sensitive biological system and representing a suspension containing predominantly 146S and 75S immunogenic components of FMDV type A.

2. Avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007, preferably obtained in a transplantable cell culture of animal origin and representing a suspension containing predominantly 146S and 75S immunogenic components of FMDV type A.

3. Avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007, obtained preferably in a continuous cell culture of VNK-21 and representing a suspension containing predominantly 146S and 75S immunogenic components of FMDV type A.

4. Avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007-DEP, obtained preferably in a continuous cell culture VNK-21 and representing a suspension containing predominantly 146S and 75S immunogenic components of FMDV type A in an amount of at least 3, 0 mcg in 1 cm 3 of the finished product.

5. As a targeted additive, the vaccine contains an adjuvant-sorbent of GOA.

6. GOA preferably in the amount of 11000.0÷15000.0 μg per 1 cm 3 of the finished product.

7. As a targeted additive, the vaccine contains the adjuvant saponin.

8. Saponin preferably in the amount of 500.0 ÷ 1500.0 μg per 1 cm 3 of the finished product.

9. As a targeted supplement, the vaccine contains a supporting medium.

10. Maintenance medium in an amount of up to 1,000,000.0 μg per 1 cm 3 of the finished product.

11. Avirulent and purified antigenic material from strain A no. ratio, mcg:

The proposed vaccine has a high immunogenic activity and provides reliable protection against FMDV serotype A, circulating in the countries of the Caucasus, Central Asia, the Middle and Far East.

The achievement of the technical result from the use of the invention is achieved by the fact that the antigenic material from strain A No. 2045/Kyrgyzstan/2007 of FMDV serotype A, which has high biological, antigenic and immunogenic activity in native form and after inactivation and which provides for the production of an adsorbed inactivated FMD vaccine that creates effective protection for susceptible animals against FMDV serotype A, which causes outbreaks in last years in the countries of Transcaucasia, Central Asia, the Near and Far East.

Strain A No. 2045/Kyrgyzstan/2007 is new, previously unknown. The initial virus for obtaining strain A No. 2045/Kyrgyzstan/2007 was isolated from samples of aphthous material from a sick heifer received by FGU "ARRIAH" in December 2007 from the Republic of Kyrgyzstan (expertise No. 2045 "Kyrgyzstan/07"). Production strain A No. 2045/Kyrgyzstan/2007 of FMDV type A was obtained by successive passages on sensitive hetero- and homologous cell cultures.

The resulting strain was deposited on July 7, 2009 in the All-Russian State Collection of Microorganism Strains Used in Veterinary Medicine and Animal Husbandry, Federal State Institution "All-Russian State Center for Quality and Standardization of Medicines for Animals and Feed" (FGU "VGNKI") under the registration number (link): production strain A No. 2045/Kyrgyzstan/2007-DEP of FMDV type A.

Strain A No. 2045/Kyrgyzstan/2007-DEP of FMDV type A is characterized by the following features and properties.

Morphological properties

Strain A No. 2045/Kyrgyzstan/2007-DEP belongs to the family Picornaviridae, genus Aphtovirus, serotype A and has morphological features characteristic of the FMD pathogen: the shape of the virion is icosahedral, the size is 23-25 ​​nm. A virion consists of an RNA molecule enclosed in a protein coat. The protein shell consists of 32 capsomeres arranged in cubic symmetry.

Antigenic properties

According to its antigenic properties, FMD virus strain A No. 2045/Kyrgyzstan/2007 belongs to serotype A. The virus is stably neutralized by homologous antiserum. The virus does not show hemagglutinating activity (HA activity). In sick animals, antibodies are formed in the blood serum, which are detected in the enzyme-linked immunosorbent assay (ELISA) and microneutralization reaction (RMN). When cattle are immunized with a vaccine from an inactivated virus, it induces the formation of specific antibodies detected in ELISA, RDP and RMN.

During hyperimmunization of guinea pigs, the concentrated antigen from the inactivated foot-and-mouth disease virus type A, strain A No. 2045/Kyrgyzstan/2007 induces the formation of virus-specific and virus-neutralizing antibodies detected in RSK at dilutions of 1:80÷1:160 and in ELISA at dilutions of 1:10000÷1: 12000.

The antigenic relationship of strain A No. 2045/Kyrgyzstan/2007 of FMDV type A with the production strain of FMDV A 22 No. 550 and strains previously isolated on the Asian continent was studied in the RSK, ELISA and RMN.

The results of studies in the RSC are presented in Table 1. Antigenic relatedness (R) of strain A No. 2045 / Kyrgyzstan / 2007 with other FMD viruses of serotype A was 8% for A / Iran / 97, A 22 / Iraq 24/64 - 28%, A/Iran/05 - 68%, A/Turkey/06 - 66%, A 22 No. 550 - 14%.

The results of studies in the RMN are presented in table 2. Antigenic correspondence (r 1) was for A / Iran / 97 - 0.125, A 22 No. 550 - 0.17, A 22 / Iraq 24/64 - 0.6, A / Turkey / 06 - 0.5. With an r 1 value >0.3, the field isolate and the production strain are closely related, and a vaccine from the production strain will protect against the epizootic virus, with an r value of 1<0,3 полевой изолят отличается от производственного штамма, и вакцина из данного штамма не защищает от эпизоотического вируса .

The results of studies in the ELISA are presented in table 3. Antigenic correspondence (r 1) was for A 22 No. 550 - 0.24, A 22 / Iraq 24/64 - 0.24, A / Iran / 97 - 0.19, A / Turkey/06 - 0.5. When the value of r 1 equal to 0.4÷1.0, the field isolate and the production strain are in close antigenic relationship; with a value of r 1 0.2÷0.39, the field isolate is antigenically related to the production strain, the vaccine from the production strain can be used if no more related strain is found, and provided that the animals are immunized more than 1 time; with value r 1<0,2 полевой изолят отличается от производственного штамма, вакцина из которого не приемлема для защиты от заражения полевым вирусом .

Molecular genetic characterization

The primary structure of the VP 1 gene of FMDV type A, strain A No. 2045/Kyrgyzstan/2007 was determined by nucleotide sequencing and the primary structure of the VP 1 protein was derived. Comparative analysis of nucleotide sequences showed that strain A No. 2045/Kyrgyzstan/2007 close phylogenetic relationship with strains A/Iran/05 and A/Turkey/06 (percentage of nucleotide differences 3.82÷4.3 and 4.46÷5.57, respectively) and differs significantly from vaccine strain A 22 No. 550. In addition , it differs from strains of this type of virus isolated in 1998 in Armenia and Turkey, as well as in Georgia and Iran in 1999 (see dendrogram). The degree of nucleotide differences in the sequences of strain A No. 2045/Kyrgyzstan/2007 of foot-and-mouth disease virus type A with strains of foot-and-mouth disease virus of serological type A was: with strain A 22 No. 550 - 18.15%, with strain A No. 1707 "Armenia-98" - 17, 68%, with strain A/Turkey/98 - 17.2%, with strain A (Georgia) 1999/ No. 1721 - 18.79%, with strain A/Iran/99 - 17.68%.

Thus, phylogenetic analysis showed that strain A No. 2045/Kyrgyzstan/2007 of FMDV type A belongs to a new genetic line of FMDV serotype A.

Biotechnological characteristics

Strain A No. 2045/Kyrgyzstan/2007 is reproduced in a monolayer culture of porcine kidney (SP) cells, continuous cell cultures PSGK-30, VNK-21 and IB-RS-2, and within 20÷24 hours of incubation, the virus yield in these cell cultures reaches values ​​from 6.33 to 8.25 lg TCD 50 -/cm 3 (table 4). With massive infection (1÷10 TCD/cell) causes CPP after 5 hours. It retains its original characteristics when passaged in cell cultures for 10 passages (observation period).

Chemo- and genotaxonomic characteristics

Strain A No. 2045/Kyrgyzstan/2007 of FMDV type A is an RNA-containing virus with a molecular weight of 7×10 6 D.

The nucleic acid is represented by a single-stranded linear molecule with a molecular weight of 2.8×10 6 D. The virion has a protein coat consisting of four main proteins VP 1 , VP 2 , VP 3 and VP 4 . The lipoprotein membrane is absent.

The main antigenic protein is VP 1 . The virion contains approximately 31.5% RNA and 68.5% protein. Virion RNA is infectious and is involved in the formation of precursor proteins in infected cells. The precursors, in turn, are cleaved to form more stable structural and non-structural virus polypeptides. Of the 8 non-structural polypeptides that accumulate in infected cells, one (BP 56a) is an RNA-dependent RNA polymerase involved in RNA replication of new virions.

Physical properties

The weight of the virion is 8.4×10 -18 g. The sedimentation coefficient of 146S in the sucrose gradient. Buoyant density 1.45 g/cm 3 .

Resistance to external factors

Strain A No. 2045/Kyrgyzstan/2007 is resistant to ether, chloroform, freon, acetone and other organic solvents and detergents. Most stable at pH 7.2÷7.6. Shifts in pH, both acidic and alkaline, lead to virus inactivation.

Sensitive to formaldehyde, UV radiation, radiation, high temperatures.

Additional features and properties

Immunogenic activity - immunogenic as part of an inactivated vaccine.

Reactogenicity - does not possess reactogenic properties.

Pathogenicity - pathogenic for artiodactyl animals, newborn mice, guinea pigs.

Virulence - Virulent to naturally susceptible animals by contact, aerosol and parenteral infection.

Stability - retains the original biological properties when passaged in sensitive biological systems for 10 passages.

Strain A No. 2045/Kyrgyzstan/2007 of FMDV type A exhibits high biological, antigenic and immunogenic activity both in native form and after inactivation.

Based on the data obtained, it can be argued that strain A No. 2045/Kyrgyzstan/2007, according to antigenic and immunological spectra, is an original, taxonomically new, previously unknown variant of FMDV type A.

To reduce its epizootic danger, timely vaccination of newly emerging foci of the disease is necessary, which requires topical highly immunogenic vaccines.

The analysis of the prior art carried out by the Applicant, including searching through patent and scientific and technical sources of information, and identifying sources containing information about analogues of the proposed invention, made it possible to establish that the Applicant did not find sources characterized by features that are identical (identical) to all the features of the proposed invention. The definition from the list of identified analogues of the prototype, as the analogue closest in terms of the totality of features, made it possible to establish the totality of the distinguishing features of the proposed vaccine, which are significant in relation to the technical result perceived by the applicant, as set forth in an independent claim.

Therefore, the claimed vaccine corresponds to the level of patentability "novelty".

To verify the compliance of the proposed vaccine with the patentability condition "inventive step", an additional search for known solutions was carried out to identify the features included in the distinctive part of the independent claim. The search results showed that the proposed solution does not explicitly follow for a specialist from the prior art set forth in the corresponding section of the description (no solutions have been identified that have features that coincide with the distinguishing features of the proposed invention), and the effect of the transformations provided for by the essential features of the proposed vaccine has not been identified. to achieve a technical result.

Therefore, the proposed vaccine meets the condition of patentability "inventive step".

The essence of the invention is explained in the sequence listings, in which:

SEQ ID NO:1 represents the nucleotide sequence of the VP 1 protein gene of strain A No. 2045/Kyrgyzstan/2007 of FMDV type A.

SEQ ID NO:2 amino acid sequence of the VP 1 protein of strain A No. 2045/Kyrgyzstan/2007 of FMDV type A.

The essence of the invention is illustrated on a dendrogram reflecting the phylogenetic relationships of strain A No. 2045/Kyrgyzstan/2007 of FMDV serological type A with epizootic and vaccine strains of FMDV type A. The dendrogram is based on a comparison of the complete nucleotide sequences of the VP 1 gene.

The essence of the invention is illustrated by examples of its use, which do not limit the scope of the invention.

Strain A No. 2045/Kyrgyzstan/2007 FMDV type A was isolated from the field material received by the Federal State Institution "ARRIAH" in the form of aphthae epithelium from cattle suspected of having FMD during laboratory diagnosis of this disease and its differentiation from other vesicular diseases. During the isolation of the virus, a complex of biological, virological and biochemical methods was used.

Biological and virological methods included the inoculation of the material of the field cattle isolate and subsequent adaptation of the virus to cultures of primary trypsinized and transplanted cell lines. Cell cultures SP, PSGK-30, IB-RS-2, and VNK-21 were used. Primary and continuous cell cultures for setting up a bioassay were grown on appropriate nutrient media under stationary conditions in vials with a capacity of 50÷100 cm 3 , washed from the growth medium and infected with a 10% suspension of aphthous material (multiplicity of infection was 1÷10 in Hank's solution with 0.5% HLA and standard antibiotics. To remove microflora and ballast cell components, the suspension was treated with chloroform in a ratio of 1:10. After a 30-minute incubation at 37°C, 5÷10 cm 3 of the supporting medium were added to the flasks and incubated at 37°C until the appearance of the CPE of the virus. In the presence of CPD (rounding of cells, increase in their optical density, degeneration and separation of cells from glass), the vials were subjected to freezing-thawing, purification of the cell suspension with chloroform, and centrifugation at 3000g for 15 minutes. The resulting virus-containing material was used for subsequent passages and research in RSK and ELISA for the presence of viral antigen, using a set of commercial type-specific sera and sera stored in the Museum of strains of the Federal State Institution "ARRIAH".

The results of virus adaptation to various cell cultures are presented in Table 4.

The data shown in table 4 indicate a good adaptive activity of strain A No. 2045/Kyrgyzstan/2007 FMDV type A to used cell cultures.

The virus isolated using the above methods was examined in the RSK with a set of diagnostic kits for all types of foot-and-mouth disease virus in order to identify the type affiliation.

The results of virus typing in RSCs are presented in Table 5.

The results shown in Table 5 indicate that the isolated virus belongs to type A.

The vaccine against FMD type A, inactivated sorbed, is prepared from virus strain A No. 2045/Kyrgyzstan/2007, grown in a suspension culture of VNK-21 cells. Earl's solution without serum with the addition of FGMS, GBCS and antibiotics at pH 7.4÷7.8 is used as a supporting medium. The cell culture is infected with the virus at the rate of 0.001÷0.05 TCD 50 per cell.

The cultivation of the virus is carried out at a temperature of 36÷37°C. After 11÷13 hours of incubation, living and dead cells are counted by staining with trypan blue. If the number of living cells is 15÷20%, then the incubation is continued for another 2÷3 hours. When the number of dead cells reaches 90÷95%, the cultivation is stopped and the virus-containing suspension is controlled for sterility and the content of 146S and 75S components. The amount of 146S+75S components in the suspension should be at least 0.5 µg/cm 3 .

Tables 6, 7, 8 and 9 show the results of cultivating strains A No. 2045/Kyrgyzstan/2007, A (Georgia) 1999/ No. 1721-DEP, A No. 1707 "Armenia-98-DEP" and A 22 No. 550 of FMDV type A, from which it follows that the cultural properties of the listed strains differ significantly in two studied parameters:

1) strain A no. 2045/Kyrgyzstan/2007 has a shorter reproduction period (12.5 hours) compared with strain A (Georgia) 1999/ no. 1721-DEP and A no. hours);

2) strains A No. 2045/Kyrgyzstan/2007 and A 22 No. 550 give a higher yield of immunogenic components (90.1% and 89.9%, respectively) compared to strains A (Georgia) 1999/ No. 1721-DEP and A No. 1707 "Armenia-98-DEP" (61.5% and 58.1% respectively).

Immediately after the end of the virus reproduction cycle, without stopping thermostatting, a 15÷20% AEEI solution, acidified with glacial acetic acid to pH 8.0÷8.5, is added to the virus-containing suspension. The final concentration of AEEI in the virus-containing suspension should be 0.025÷0.05%. The inactivation of the infectivity of the virus is carried out for 12÷24 hours at 36÷37°C and pH 7.2÷7.6 with stirring after 5÷6 hours for 3÷5 minutes. The AEEI residue is neutralized by adding sodium thiosulfate.

A 10% PHMG solution is added to the warm suspension to a concentration of 0.005÷0.007% to flocculate ballast impurities and inactivate possible contaminants. Flocculated ballast impurities are subjected to sedimentation followed by decantation.

Table 10 shows the results of studies on the effect of inactivation and purification of antigenic material using AEEI and PHMG on the immunogenic (146S and 75S) components of FMDV strains A No. 2045/Kyrgyzstan/2007 and A (Georgia) 1999/ No. 1721-DEP.

The data presented in Table 10 indicate that the immunogenic components of the foot-and-mouth disease virus strain A No. 2045/Kyrgyzstan/2007 are not inferior in resistance to inactivation and purification under production conditions to the foot-and-mouth disease virus strain A (Georgia) 1999/ No. 1721-DEP. Of particular importance is the fact that during the inactivation and purification of the virus from strain A no. 2045/Kyrgyzstan/2007, there were no significant changes in the amount of 146S and 75S components obtained during virus reproduction.

The resulting antigen is controlled for avirulence, the content of virus-specific protein, 146S and 75S components of the virus and sterility. The required concentration of 146S and 75S components in the inoculation dose of the adsorbed vaccine is obtained by concentrating the GOA antigen.

The estimated volume of GOA 3% concentration is added to the cooled antigen suspension with the stirrer running. Stirring is carried out for 30 minutes. After sedimentation of GOA, the calculated volume of the remaining suspension is drained. The final concentration of GOA should be within 1.62 ± 0.488 R<0,01 мг/см 3 n=10, а концентрация 146S и 75S компонентов вируса ящура, по меньшей мере, 3,0 мкг/см 3 готового препарата. Затем в суспензию добавляют дополнительно 10% раствор сапонина до конечной концентрации 0,05÷0,15%, что соответствует 500,0÷1500,0 мкг сапонина в 1 см 3 готовой вакцины.

The resulting vaccine is packaged in glass vials and its sterility is controlled in accordance with GOST 28085.

Avirulence and harmlessness of the vaccine is tested on 5 heads of cattle by administering the vaccine first under the tongue mucosa at a dose of 2.0 cm 3 and then subcutaneously at a dose of 10.0 cm 3 . The clinical condition of the animals is monitored for 10 days. An avirulent, harmless and sterile vaccine is tested for immunogenic activity in cattle or guinea pigs.

The resulting vaccine is a light yellow liquid with a loose white precipitate of the sorbent, which forms at the bottom of the vial during storage and easily breaks into a homogeneous suspension when shaken.

The optimal component composition of the obtained adsorbate vaccine against FMD type A is shown in Table 11.

Avirulence and harmlessness of the vaccine was tested on 5 heads of cattle. The drug was administered to each animal under the mucous membrane of the tongue at a dose of 2.0 cm 3 and then subcutaneously at a dose of 10.0 cm 3 . The clinical condition of the animals was monitored for 10 days.

At the end of the control for avirulence and harmlessness, the vaccine was tested for immunogenic activity. The test was carried out on 4 heads of calves weighing 70÷80 kg. The adsorbate vaccine was injected subcutaneously at a dose of 2.0 cm 3 . The control infection of calves was carried out with a homologous strain of foot-and-mouth disease virus on the 28th day after vaccination (WPV). The research results are presented in table 12.

The data presented in table 12 showed that 1 calf out of 4 fell ill, 3 were protected from the generalization of the process on day 28 after vaccination. The level of humoral immunity in vaccinated animals was 4.81±0.53 log 2 .

The adsorbate vaccine against foot and mouth disease type A was tested, manufactured as described in example 2, and containing, μg:

Avirulence and harmlessness of the resulting vaccine was tested on 5 heads of cattle. The drug was administered to each animal under the mucous membrane of the tongue at a dose of 2.0 cm 3 and then subcutaneously at a dose of 10.0 cm 3 . The clinical condition of the animals was monitored for 10 days after infection.

At the end of the control for avirulence and harmlessness, the vaccine was tested for immunogenic activity. The test was carried out on 5 heads of cattle weighing 250÷300 kg and 5 heads of calves weighing 70÷80 kg. The adsorbate vaccine was injected subcutaneously at a dose of 2.0 cm 3 . Control infection was carried out with a homologous strain of FMDV at 28 days post-vaccination (DPV) for calves and at 90 days for bulls. The research results are presented in tables 13÷14.

The data presented in tables 13÷14 showed that 1 bull out of 5 fell ill, and 4 were protected from the generalization of the process on the 90th day after vaccination, and all 5 heads of calves were protected from the generalization of the process on the 28th day after vaccination. The level of humoral immunity in vaccinated animals at the time of infection was 5.00±0.27 log 2 for bulls and 6.20±0.63 log 2 for calves, which indicates the effectiveness of the tested vaccine.

Thus, the above information indicates that the following set of conditions is met when using the proposed vaccine:

vaccine against FMD type A inactivated adsorbed, embodying the present invention, is intended for use in agriculture, namely in veterinary virology and biotechnology;

For the proposed invention in the form as it is described in the independent claim, the possibility of its implementation using the means and methods given in the application or known before the priority date is confirmed;

FMD vaccine type A adsorbed inactivated, made from strain A No. 2045/Kyrgyzstan/2007 in accordance with the invention, has a high immunogenic activity and is able to provide effective protection of susceptible animals against epizootic FMDV type A, circulating in the countries of the Caucasus, Central Asia, Near and Far East.

Sources of information

1. Bojko A. A. Declaration sur I apparition en URSS d un type virus aphteux different des souches de type A anterieurement etudiees dans le Pays. BOIE, 1965, 64, v. 2. - P. 1075-1077.

2. Pepep X. Foot and mouth disease: per. from German / G.A. Surkova; ed. and with preface. P.V. Painter. - M.: Kolos, 1971. - 432 p.

3. Foot and mouth disease: monograph / A.N. Burdov, A.I. Dudnikov, P.V. Malyarets [i dr.]. - M.: Agropromizdat, 1990. - 320 p.

4. Viral animal diseases / V.R. Syurin, A.Ya. Samuylenko, B.V. Solovyov [i dr.]. - M.: VNIITIBP, 1998. - S.532-548.

5. Interim instructions for the manufacture and control of FMD concentrated aluminum hydroxide formol vaccine from lapinized A 22 virus. Approved by the GUV USSR on March 25, 1971.

6. Industrial regulation for the production of vaccines against foot-and-mouth disease types A, O, C, Asia-1, Sat-1, Sat-2 and Sat-3 inactivated sorbed mono- and polyvalent (from a virus grown in VNK-21 cells): approved . Director of FGU “ARRIAH” on 11.09.2009. - Vladimir, 2009. - 216 p.

7. Industrial regulation for the production of the FMD vaccine inactivated emulsion mono- and polyvalent for the prevention of foot-and-mouth disease in pigs (from a virus grown in VNK-21 cells): approved. Director of FGU “ARRIAH” on 11.09.2009. - Vladimir, 2009. - 240 p.

8. Pat. Russian Federation 2294760, IPC A61K 39/135, Inactivated sorbed vaccine against FMD type A / V.V. Mikhalishin, T.N. Lezova, A.V. Shcherbakov, D.V. Mikhalishin, N.S. Mamkov; FGBU "ARRIAH" - Appl. December 21, 2004; publ. 06/10/2006 (prototype).

9. Pat. RF 2140452, IPC A61K 39/135, Strain N 1707 "ARMENIA-98" of FMDV type A for the manufacture of diagnostic and vaccine preparations / V.M. Zakharov, V.K. Spirin, A.I. Gritsenko [and others]; FGBU "ARRIAH" - Appl. 03/15/1999; publ. October 27, 1999

10. Pat. RF 2242513, IPC A61K 39/135, Strain A (GEORGIA) 1999/ No. 1721 of FMDV type A for the manufacture of diagnostic and vaccine preparations; FGU "ARRIAH" - Appl. 11/24/2003; publ. December 20, 2004

11. Foot-and-mouth disease. Sitiation worldwide and major epidemiological events in 2005-2006/ K. Sumption, J. Lubroth, T. Murrai, S. De la Rocque// Empress/Focus on. - 2007. - No. 1.

12. Pat. Russian Federation 594771, IPC A61K 39/12, Agent for inactivating viruses in the manufacture of antiviral drugs / N.A. Ulupov, A.I. Dudnikov, P.A. Gembitsky, D.S. Zhuk; VNIYA - Appl. 05/07/1973; publ. 07/07/93

13. Pat. Russian Federation 2054039, IPC A61K 39/135, Method for purification and sterilization of cultural FMD virus / T.N. Lezova, N.A. Ulupov, V.V. Borisov, V.V. Mikhalishin, A.I. Dudnikov, P.A. Gembitsky; VNIYA - Appl. 02/07/1992; publ. February 10, 1996

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CLAIM

1. FMD vaccine type A inactivated sorbed, containing the active substance and target additives, characterized in that as the active substance it contains avirulent and purified antigenic material from the Aphtae epizooticae virus strain, fam. Picornaviridae, genus Aphtovirus, serotype A, deposited in the collection of the Federal State Institution "VGNKI" under the registration number production strain of FMD virus serotype A No. 2045/Kyrgyzstan/2007-DEP, in an effective amount.

2. The vaccine according to claim 1, characterized in that it contains avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007-DEP, obtained in a sensitive biological system and representing a suspension containing predominantly 146S and 75S immunogenic components of FMDV type A.

3. The vaccine according to claim 2, characterized in that it contains avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007-DEP, obtained preferably in a transplanted cell culture of animal origin and representing a suspension containing mainly 146S and 75S immunogenic components of FMDV type A.

4. The vaccine according to claim 3, characterized in that it contains avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007-DEP, obtained preferably in a continuous cell culture VNK-21 and representing a suspension containing mainly 146S and 75S immunogenic components of FMDV type A.

5. The vaccine according to claim 4, characterized in that it contains avirulent and purified antigenic material from strain A No. 2045/Kyrgyzstan/2007-DEP, preferably obtained in a continuous cell culture of VNK-21 and representing a suspension containing mainly 146S and 75S immunogenic components of foot-and-mouth disease virus type A in an amount of at least 3.0 μg per 1 cm 3 of the finished product.

6. The vaccine according to claim 1, characterized in that it contains an adjuvant-sorbent aluminum hydroxide (GOA) as a target additive.

7. The vaccine according to claim 6, characterized in that it contains GOA, preferably in the amount of 11000.0÷15000.0 μg per 1 cm 3 of the finished product.

8. The vaccine according to claim 1, characterized in that it contains a saponin adjuvant as a target additive.

9. The vaccine according to claim 8, characterized in that it contains the adjuvant saponin, preferably in the amount of 500.0÷1500.0 μg per 1 cm 3 of the finished product.

10. The vaccine according to claim 1, characterized in that it contains a supporting medium as a target additive.

11. The vaccine according to claim 10, characterized in that it contains a supporting medium in an amount of up to 1,000,000.0 μg per 1 cm 3 of the finished product.

12. The vaccine according to any one of claims 1-11, characterized in that it contains avirulent and purified antigenic material from strain A No. predominantly 146S and 75S immunogenic components of foot-and-mouth disease virus type A, GOA, saponin and supporting medium in the ratio, mcg.

Approved

Head Office

Ministry of Veterinary

agriculture of the USSR

TEMPORARY INSTRUCTION

ON THE USE OF FMD VACCINE

TYPE A, TYPE O, TYPE C, TYPE ASIA-1

(FROM VIRUS GROWN IN BHK-21 CELLS)

1. General Provisions

1.1. The monovalent vaccine is intended for prophylactic immunization of cattle, yaks, buffaloes, sheep and goats against FMD caused by one of the serotypes A, O, C, Asia-1.

1.2. The monovalent FMD vaccine is made from foot-and-mouth disease virus grown in a suspension of VNK-21/2-17 cells adsorbed on aluminum hydroxide, inactivated by formaldehyde with the addition of saponin.

For the manufacture of the drug use one of the following strains of the virus

foot and mouth disease: A, O, C, Asia-1.

22 1

In appearance, it is a light yellow liquid, with a loose, easily broken white precipitate that forms at the bottom of the vial (during storage) when shaken.

1.3. Vials with the vaccine must be hermetically sealed and labeled with the name of the manufacturer and its trademark, the name of the drug, the amount of the drug in the vial, the serial number and state control, the date of manufacture (month, year), expiration date, storage conditions, designation of technical conditions, vaccine dose.

1.4. The vaccine is suitable for use within 12 months. after manufacture, subject to storage and transportation conditions.

FMD vaccine monovalent type A, type O, type C, type Asia-1

at the manufacturing plant, in the organizations of Soyuzglavzoovetsnabprom and at

the consumer is stored in a dry, dark and closed room at a temperature of 4 -

8 °C. A vaccine that has been frozen and, as a result, has lost its

colloidal properties and the ability to give when shaking the vial

homogeneous stability of suspension of aluminum hydroxide, to use

unsuitable and subject to rejection.

1.5. If there are foreign impurities, flakes, mold in the vaccine, if the integrity of the vial is broken, if the label is missing, and if the drug is not used on the day the vial is opened, the vaccine must be destroyed.

2. The method of application of the vaccine

2.1. The vaccine is administered strictly subcutaneously to cattle, yaks and buffaloes in the area of ​​the middle third of the neck, to sheep and goats - from the inside of the thigh in the doses indicated on the vial labels.

2.2. When carrying out vaccinations, it is necessary to follow the rules of asepsis and antisepsis.

At the injection site, the skin surface is thoroughly disinfected with a 70% solution of ethyl alcohol. Syringes and needles are sterilized by boiling before use; a separate needle is used for each animal.

3. Procedure for the use of vaccines

3.1. The vaccine is used for prophylactic purposes and involuntarily in a disadvantaged area and in a zone threatened by foot and mouth disease when specifying the type of virus.

3.2. In farms where the vaccine of this serotype has not been previously used, all animals are immunized, regardless of age, as well as young animals born within a month.

Revaccination is carried out after 2 months, and then the adult population is immunized every 6 months. after the first vaccination, young animals up to 18 months of age - every 3 months. after the second vaccination.

3.3. Immunity in primary vaccinated animals occurs by the 21st day, and subject to the intervals between revaccination and subsequent immunizations, immunity to this serotype is maintained constantly.

3.4. Young animals born from immunized animals are vaccinated from the age of 4 months, and subsequently according to clause 3.2 of this Temporary Instruction.

4. Observation of vaccinated animals

4.1. The vaccinated animals are monitored, keeping in mind that some of these animals have a local reaction at the injection site, which manifests itself in the form of a small painful swelling, which disappears after 5-10 days.

The general reaction of the body can manifest itself in the form of a short-term increase in body temperature by 1 - 2 ° C for 1 - 3 days.

4.2. Animals in which an allergic reaction is observed during revaccination are injected with antihistamines (diphenhydramine, adrenaline, atropine sulfate, caffeine, etc.).

4.3. In case of complications that have arisen after the introduction of the vaccine, the All-Union Order of the Red Banner of Labor is immediately reported to the State Scientific and Control Institute of Veterinary Preparations of the USSR Ministry of Agriculture (123022, Moscow, D-22, Zvenigorodskoye shosse, 5) and the biological enterprise that manufactured the vaccine.

In accordance with the directive of the Main Directorate of Veterinary Medicine of the Ministry of Agriculture of the USSR dated 09/08/1982 "On the procedure for submitting a complaint for biological products", at least 3 vials of vaccine placed in a thermos with a temperature of 2 - 8 ° C are simultaneously sent to VGNKI by courier, and an accompanying document in which indicate the number of vaccinated and diseased animals, the nature of complications, the result of virus typing, the epizootic situation for foot-and-mouth disease and other data characterizing the course of the disease.

The Association assists in the provision of services in the sale of timber: at competitive prices on an ongoing basis. Timber products of excellent quality.