RU2465313C2 - Chicken coccidiosis (eumeriosis) vaccine - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: presented vaccine contains an antigen in the form of a suspension of mixed sporulated oocysts of the coccidiosis strains deposited in the collection of the State Scientific Institution All-Russian Scientific Research Veterinary Institution of Poultry Industry of Russian Agricultural Academy - the coccidiosis strain Eimeria acervulina L-2-15 KE No.2, the coccidiosis strain Eimeria maxima L-3-2 KE No.3, the coccidiosis strain Eimeria necatrix L-4-18 KE No.4, the coccidiosis strain Eimeria tenella L-1-23 KE No.1 taken in relations 3000:500:1000:1000 of sporulated oocysts in one immunising dose; a preserving agent is 1% chloramine.

EFFECT: invention provides producing immunity to four types of eumeria in chickens, has a long shelf life.

4 tbl, 4 ex

Description

The invention relates to veterinary parasitology and biotechnology, and in particular to veterinary preparations for the prevention of coccidiosis (eimeriosis) chickens.

Vaccination of birds against coccidiosis is an alternative way to prevent this parasitosis using anti-coccidia drugs (chemical or artificial antibiotics). The latter method is currently the most widely used in poultry farming. However, due to the fact that existing trends in obtaining environmentally friendly food products suggest a gradual departure from the use of not only antibiotics, but also chemotherapeutic agents in poultry feeds, the use of vaccines will probably become the main way to prevent coccidiosis in poultry farming.

As you know, immunity in coccidiosis is developed only after the parasite passes a certain development cycle in the cells of the intestinal mucous membrane of chickens and lasts for a short time without repeated infections. The main method of vaccine use is the oral administration of strictly dosed amounts of exogenous stages of the development of the parasite (oocysts), which cause the formation of immunity to coccidiosis without the clinical manifestation of the disease. At the same time, parasites go through a full development cycle, and vaccinated chickens secrete a small amount of oocysts into the environment, which, when pecked, enter the chickens again, which leads to maintaining immunity in the herd at a sufficient level during the entire period of growing the bird.

Known industrial vaccine against coccidiosis, developed by Edgar (USA) and sold in 1952 under the name "DM ^® Cecal coccidiosis Vaccine", which contained live oocysts of the same species - Eimeria tenella. (one)

Currently, two types of foreign live vaccines are also known - non-attenuated (Coccivac ^® , Immucox ^® , Nobilis ^® COX ATM, VAC M ^® ) and attenuated (Paracox ^® , Livacox ^® ) (2). All vaccines contain live exogenous stages of parasites (oocysts) in certain quantities. There is also a Coxabic ^® subunit vaccine.

However, these drugs are not always effective when used in domestic poultry farms.

There is a known coccidia culture for the immunochemistry of chickens eimeriosis, which is the prototype of the claimed invention and contains titrated immunogenic doses of several types of eimeria oocysts, which is used with the use of an anti-coccidia drug for 20 days. (3, 4).

This coccidia culture is introduced with food and allows the bird to be immune to three species of eimeria (Eimeria tenella, Eimeria acervulina, Eimeria maxima).

Tense immunity is formed in the bird, the safety of the livestock, the yield of young business women and the average daily increase are increased, and the cost of feed is reduced.

However, in this case, immunity is formed only against the three types of eimeria included in the culture, and if another pathogenic species is widespread in the economy, then it will inevitably cause an outbreak of the disease, since immunity to eimeria is not specific. We also note that at present, the strain Eimeria necatrix is widely used in poultry farms of the Russian Federation.

The well-known drug also has a short shelf life (3 months), the restriction in use is only with food, contains formaldehyde as a preservative. All this reduces the effectiveness of the prevention of coccidiosis in modern conditions.

The problem to be solved within the framework of the claimed invention is the creation of a vaccine for the immunoprophylaxis of coccidiosis (eimeriosis) chickens with high immunogenic properties, environmentally friendly and economical, with an extended shelf life, more technological to use.

The problem is solved by creating a vaccine containing, as an antigen, a suspension of a mixture of sporulated oocysts of coccidiosis pathogen strains deposited in the collection of GNU VNIVIP of the Russian Agricultural Academy -

a strain of the causative agent of coccidiosis Eimeria acervulina L-2-15 KE No. 2,

strain of the causative agent of coccidiosis Eimeria maxima L-3-2 KE No. 3,

a strain of the causative agent of coccidiosis Eimeria necatrix L-4-18 KE No. 4,

a strain of the causative agent of coccidiosis Eimeria tenella L-1-23 KE No. 1 in a ratio of 3000: 500: 1000: 1000 sporulated oocysts in one immunizing dose, and as a preservative - 1% aqueous solution of chloramine.

The high immunogenic properties of the claimed vaccine are due to both the high immunogenic properties of the strains used and the expansion of the species composition of coccidia contained in it.

For immunization of birds, specially selected coccidia strains are used, which have low virulence and high immunogenicity, in doses that do not cause clinical disease of coccidiosis. The above strains possess such properties.

An increase in the shelf life of the vaccine up to 6 months is achieved through the use of chloramine or ecocide in the indicated concentration.

The use of the claimed vaccines for the prevention of coccidiosis (eimeriosis) of chickens leads to the abandonment of the use of anti-coccidia drugs after immunization of the livestock, which, in turn, has a positive effect on the environmental safety of food.

The inventive vaccine is more technologically advanced and convenient to use, since it can be used not only with food, but also with a method of drinking, which also contributes to the uniform development of immunity in the entire livestock.

The claimed vaccine is made from production strains of four types of coccidia deposited in the collection of GNU VNIVIP of the Russian Agricultural Academy -

a strain of the causative agent of coccidiosis Eimeria acervulina L-2-15 KE No. 2,

strain of the causative agent of coccidiosis Eimeria maxima L-3-2 KE No. 3,

a strain of the causative agent of coccidiosis Eimeria necatrix L-4-18 KE No. 4,

a strain of the causative agent of coccidiosis Eimeria tenella L-1-23 KE No. 1, which are propagated in 14-day-old chickens, free from eimeria.

In appearance, the vaccine is a homogeneous suspension of a grayish-orange color. During storage, it precipitates, which easily breaks when shaken into a homogeneous suspension.

The strains are maintained by passivation on 14-day-old coccidia-free chickens at least once every 6 months.

Characterization of the production strain L-2-15 Eimeria acervulina KE No. 2 .

Systematic position. Strain L-2-15 E. acervulina CE No. 2 belongs to the genus Eimeria, the family Eimeriidae, and the order Coccidiida.

Morphological characteristics. The oocysts are ovoid in shape, 14.0-23.0 microns long, 13.0-18.0 microns wide (average 18.7-14.6 microns).

Biological properties. The prepatent period for the development of the parasite is 96 hours. The place of the endogenous development of the parasite in the intestines of invaded chickens is the duodenum 12 and the upper third of the small intestine.

Clinical and pathological signs. Characteristic for this species.

Virulent properties. When 15-day-old chickens are infected with oocysts that have been stored for no more than 2 months at a dose of 6 million oocysts per head, mortality is at least 30%.

Immunogenic properties. Infection of chickens with oocysts causes the formation of specific immunity (humoral and cellular).

Cultural and biochemical properties. Strain L-2-15 E. acervulina KE No. 2 is not cultivated on nutrient media. It is maintained by passaging on chickens, causing characteristic lesions in the intestine after 72 hours with the release of oocysts, starting from the 96th hour after infection.

The strain is sensitive to anticoccidic drugs - amprolium, nicarbazine, pharmacocide, chemicalcocide, clinacoxid, to all ionophore antibiotics.

Specificity. Strain L-2-15 E. Acervulina CE No. 2 is specific only for chickens (5).

Characterization of strain L-2-3 Eimeria maxima (causative agent of coccidiosis) CE No. 3

Systematic position. Strain L-3-2 E. maxima belongs to the genus Eimeria, the family Eimeriidae, and the order Coccidiida.

Morphological characteristics. Ovoid oocysts, 26.0-36.0 microns long, 18.0-27.0 microns wide (average - 30.0-22.0 microns).

Biological properties. The prepatent period for the development of the parasite is 126 hours. The place of endogenous development of the parasite in the intestines of invaded chickens is the entire small intestine.

Clinical and pathological signs. Characteristic for this species.

Virulent properties. When 15-day-old chickens are infected with oocysts that have been stored for no more than 2 months at a dose of 3 million oocysts per head, the mortality rate is at least 30%.

Immunogenic properties. Infection of chickens with oocysts causes the formation of specific immunity (humoral and cellular).

Cultural and biochemical properties. The strain is not cultured on nutrient media. It is maintained by passivation in chickens, causing characteristic lesions in the intestine after 96 hours with the release of oocysts, starting from the 126th hour after infection.

The strain is sensitive to anticoccidic drugs - amprolium, nicarbazine, pharmacocide, chemicalcocide, clinacoxid, to all ionophore antibiotics.

Specificity. Strain L-3-2 E. maxima is specific only for chickens (6).

Characterization of the production strain L-4-18 Eimeria necatrix CE No. 4

Systematic position. Strain L-4-18 of E.necatrix belongs to the genus Eimeria, the family Eimeriidae, and the order Coccidiida.

Morphological characteristics. It has elongated oval oocysts 17-26 microns long, 15-20 microns wide (average - 21.5-17.5 microns).

Biological properties. The prepatent period for the development of the parasite is 128 hours. The place of endogenous development in the intestines of invaded chickens is the middle section of the small intestine, oocysts develop in the mucosa of the blind processes.

Virulent properties. When 15-day-old chickens are infected with oocysts that have been stored for no more than 2 months at a dose of 900,000 oocysts per head, the mortality rate is at least 30%.

Immunogenic properties. Infection of chickens with oocysts causes the formation of specific immunity (humoral and cellular).

Cultural and biochemical properties. Strain L-4-18 E.necatrix is not cultured on nutrient media. It is maintained by passivation in chickens, causing characteristic lesions in the intestine after 96 hours with the release of oocysts, starting from 128 hours after infection.

The strain is sensitive to anticoccidic drugs - amprolium, nicarbazine, pharmacocide, chemicalcocide, clinacoxid, to all ionophore antibiotics.

Specificity. Strain L-4-18 of E.necatrix is specific only for chickens (7).

Characterization of the production strain L-1-23 Eimeria tenella CE No. 1

Systematic position. Strain L-1-23 of E. tenella belongs to the genus Eimeria, the family Eimeriidae, and the order Coccidiida.

Morphological characteristics. It has wide oval oocysts 19.0-26.0 microns long, 16.0-22.0 microns wide (average 22.6-18.8 microns). Schizonts of the 2nd generation has dimensions.

Biological properties. The prepatent period for the development of the parasite is 130 hours. The place of the endogenous development of the parasite in the intestines of invaded chickens is the blind processes.

Virulent properties. If 15-day-old chickens are infected with oocysts that have been stored for no more than 2 months at a dose of 800,000 oocysts per head, the mortality rate is 50% (LD ₅₀ ).

Immunogenic properties. Infection of chickens with oocysts causes the formation of specific immunity (humoral and cellular).

Cultural and biochemical properties. Strain L-1-23 E. tenella is not cultivated on nutrient media. It is maintained by passivation in chickens, causing characteristic lesions in the intestine after 72-96 hours with the release of oocysts, starting from the 130th hour after infection.

The strain is sensitive to anticoccidic drugs - amprolium, nicarbazine, pharmacocide, chemicalcocide, clinacoxid, to all ionophore antibiotics.

Specificity. Strain L-1-23 E. tenella is specific for chickens only. The parasite can go through the full development cycle also in chicken embryos (8).

All strains are sensitive to amprolium, clinocox, nicarbazine and ionophore antibiotics.

Passivation of strains.

Chickens are infected in the goiter with a syringe and a polyethylene cannula with sporulated coccidia oocysts in a dose of: 50 thousand Eimeria acervulina, 20 thousand E. maxima, 40 thousand E. necatrix, 2-3 thousand E. tenella . The volume of the inoculum is not more than cm ³ . Passages of each type of cocidia are carried out separately.

On the third day after infection, a fecal mass of chickens is examined for coccidia oocysts in order to exclude spontaneous coccidia infection in accordance with GOST 25383-82.

Litter for oocysts for chickens infected with Eimeria acervulina is collected from 5 to 8 days after infection, E. maxima from 6 to 9 days, E. necatrix from 6 to 9 days, E. tenella from 6 to 10 days. To prevent drying of the droppings during the collection period, a double layer of filter paper, abundantly wetted with a 2% solution of potassium dichromate, is laid on pallets.

Obtaining and purification of coccidia from the litter is carried out using the principle of flotation of oocysts in a saturated solution of sodium chloride. The homogenized litter mixed with water is filtered through a sieve, the filtrate obtained is precipitated in a centrifuge with a cup volume of 500 cm ³ or more for 5 minutes at 3000 rpm.

The supernatant is drained, saturated salt solution is added to the precipitate, it is thoroughly mixed and centrifuged for 10 minutes at 100 rpm. Surfaced oocysts are collected by a peristaltic pump in a vessel with water, making sure that the ratio of saturated solution of sodium chloride with oocysts and water is 1:10 (density on a densimeter 1.010-1.020). In order to maximize oocyst production, the precipitate is centrifuged three times. Then the collected oocysts are precipitated from the water in a centrifuge for 5 minutes at 3000 rpm.

Sporulation of oocysts is carried out in a 2% solution of potassium dichromate with abundant air access (using the MK-L2 microcompressor).

At a temperature of 26 ± 2 ° C for 3-5 days. The percentage of sporulation of oocysts is determined under a microscope at medium magnification.

Sporulated oocysts are stored between the passages in a 1% chloramine solution in conical flasks filled with no more than 1/3 of the volume under cotton-gauze plugs or paper caps in a refrigerator at a temperature of 4 ± 2 ° С.

In order to control production strains of coccidia, it is necessary to examine the previously mentioned properties of these strains (sizes of oocysts, prepatent period, sensitivity to coccidiostatics) after 10-15 passages.

The length and width of the oocysts are measured using a microscope with an MBI-3 brand ocular micrometer, volume x 90, size x 10. 100 sporulated oocysts of each strain are measured, average sizes are calculated. These biometric indicators should not differ significantly from the above.

The prepatent period (from the time of infection to the appearance of the first oocysts in the litter of infected chickens) is established by examining the litter of chickens infected with Eimeria acervulina - starting from 4 days, E. maxima, E. necatrix, E. tenella - starting from 5 days after infection. Fluctuations in the duration of the prepatent period should be no more than ± 5 hours.

To determine the sensitivity of coccidia strains to anti-coccidia drugs, two-week-old chickens, divided into groups of 10 animals, are infected with cocidia in a dose that causes the death of at least 50% of the chickens, and within 10 days they are given anti-coccidia preparations with food in a prophylactic dose, starting to be used the day before infection . Accounting is carried out according to the manifestation of clinical signs, mortality from coccidiosis and weight gain in experimental and control (without the use of the drug) groups.

Vaccine production includes the following steps:

- growing chickens free of coccidia;

- obtaining coccidia oocysts;

- drawing up a production series of vaccines;

- vaccine control.

1. Growing chickens free of coccidia.

Chickens intended for infection with industrial strains of coccidia and for controlling the quality of the vaccine are obtained at the age of one day from a farm that is free of infectious diseases.

They contain chickens in isolated conditions that exclude spontaneous infection with coccidia and pathogens of infectious diseases. To do this, they are grown in a separate box in metal cages, which are burned with a blowtorch before landing. When feeding, feed is used, heated for 30 minutes at 70-80 ° C, watering is carried out with boiled and chilled water. Care for this bird is carried out by a person who does not have contact with another bird and does not participate in work related to the production of the vaccine from the beginning of preparation of the premises to the transfer of the raised bird. Before infection of the birds with production strains or the start of vaccine control, a chick litter is examined for oocysts in order to exclude spontaneous coccidia infection in accordance with GOST 25383-82. In the future, only non-invasive birds are used in the work.

2. Obtaining oocysts.

Infection of chickens, collection of litter from them, isolation and sporulation of coccidia oocysts is carried out according to the method described above. Reproduction of oocysts of each type of coccidia is carried out separately.

3. Designing a vaccine production line.

Oocysts of different types of coccidia serve as the material for compiling the production series of the vaccine. One immunizing dose contains: Eimeria acervulina - 3 thousand, E.maxima - 0.5 thousand, E.necatrix - 1.0 thousand, E.tenella - 1.0 thousand sporulated oocysts.

Counting the number of oocysts of each species is carried out separately. Before counting, the flask with a suspension of oocysts is placed on a magnetic stirrer for 10 minutes.

Oocysts count in the Goryaev’s chamber (at least 3 chambers), deriving the arithmetic mean, which is multiplied by a factor of 1111. The resulting number shows the number of oocysts in 1 ml of suspension. Knowing the total volume of the suspension, the number of doses of each type is calculated.

To compile a certain number of vaccine doses, the required volume of each type of coccidia is calculated, poured into a single container and precipitated on a centrifuge at 3000 rpm for 10 minutes in assembled centrifuge glasses. The supernatant is poured, and the precipitates are resuspended in a 1% chloramine solution and poured into a sterile container, adjusted to the required volume with a 1% chloramine solution. The contents are thoroughly mixed and poured into sterile volumetric flasks.

The vaccine is poured into sterile glass bottles of 5 and 10 thousand doses with constant stirring on a magnetic stirrer, filling no more than 2/3 of the volume. The vials are closed with rubber stoppers and rolled up with aluminum caps, ensuring tightness. Labels with the name of the manufacturer and its trademark, the name of the preparation, the specific composition of the vaccine, the serial number and control, the date of manufacture (date, month, year), the number of doses in the bottle and volume (cm ³ ) are glued onto the vaccine vials. designations of service stations, storage conditions.

3.1. Vaccine control.

Each series of vaccines must be accepted (verified) by the state controller of the manufacturer.

A vaccine series is considered to be a mixture of coccidia oocysts obtained in the process of simultaneous manufacture under the same production conditions, combined in one container, packaged in glass bottles, received its number, state control number and executed in one document on the quality of the established form. To check the quality of the vaccine, the state controller makes a selection from different places in the series depending on the volume of the series:

from a series to 1500 bottles - 2% of all bottles;

from a series of 1500-3000 bottles - 1% of all bottles;

from a series of more than 3000 bottles - 0.5% of all bottles.

After checking the quality of packaging, packaging and labeling, the selected bottles of the drug are handed over to the warehouse, the selected bottles are stored in the archives of the state controller for 9 months.

Vials with the vaccine sent to the archives of the state controller are sealed and supplied with a document of the established form indicating: the name of the preparation, date of manufacture (day, month, year), batch number, state control number, date of sampling, total number of packaging units, batch size, position and signature the person who took the sample, the designation STO.

To determine the color, the presence of impurities, mold, unbroken sediment, each vaccine vial is thoroughly shaken and viewed in transmitted light, at the same time, the vials are checked for packaging strength and labeling. The drug should have the appearance of a homogeneous suspension of a grayish-orange color, without the presence of mold and not breaking lumps.

3.2. Determination of harmlessness

Equipment and materials

When conducting research apply:

- magnetic stirrer

- glass glasses with a volume of 100 cm ³ in accordance with GOST 25336

- injection syringes of the “Record” type with a capacity of 1-5 cm ³ according to GOST 18137

- chickens of 5 days of age.

Test

The test is carried out on 5-day-old chickens free of coccidia. The absence of oocysts in fecal matter is determined according to GOST 25383.

Chickens of 5 days of age are divided into two groups of 10 animals (experimental and control) and weighed. The birds of the experimental group are injected into the goiter 5 doses of the vaccine and transferred to the floor. Chickens in the control group are kept intracellularly and coccidiostatic is introduced into the feed in a prophylactic dose.

The vaccine is considered harmless if the vaccinated chickens do not show clinical signs of coccidiosis, viral and bacterial infections within 10 days. The increase in body weight in them should not differ significantly from that of chickens in the control group. It is permissible to have traces of blood in the fecal matter for 5-7 days.

In case of death of coccidiosis among vaccinated chickens of two heads or the manifestation of clinical signs of a disease of viral or bacterial etiology, the experiment is repeated on twice the number of chickens.

3.3. Determination of viability

Equipment and materials

When conducting research apply:

- biological light microscope

- centrifuge up to 3 thousand revolutions per minute

- bacteriological loops

- Goryaev’s camera

- subject glasses in accordance with GOST 9284

- glass coverslips GOST 6672

- magnetic stirrer

- homogenizer.

Test

Ten chickens of 5 days of age are injected into the goiter with one dose of the vaccine and transferred to the floor. On the sixth day, litter is collected from the chickens and an averaged sample of 100 g is taken from it. The litter sample is washed with one liter of tap water through a metal sieve with a mesh diameter of 0.05 cm. The filtrate is centrifuged at 3 thousand rpm for 5 minutes. The precipitates are combined, 200 ml of saturated sodium chloride solution are added, homogenized and centrifuged at 1 thousand rpm for 10 minutes. The peristaltic pump removes the top layer of the supernatant of 10-15 ml. The precipitate was resuspended, centrifuged and the top layer was removed. The process of removing oocysts is repeated three times. The volume of the removed liquid is measured and diluted with tap water 1:10 (if the calculation of oocysts in the Goryaev chamber is difficult, then the dilution is increased). Divorced oocysts are stirred on a magnetic stirrer for at least 10 minutes, fuel 2-3 Goryaev cameras and count the oocysts in all 225 squares of the chamber. When counting, oocysts are divided by size into three categories - small, medium, large. From the results obtained, the average value for the categories of oocysts is derived and the amount of 1 g of litter is determined by the formula:

x = a * b * 1111 * v: m, where

x - the number of oocysts in 1 g of litter

a - the average number of oocysts in the chamber of Goryaev

b - dilution coefficient

1111 - conversion factor of the number of oocysts in 1 ml

V is the volume in ml obtained by dilution

m is the weight of the litter.

Oocysts are considered viable if their total number in 1 g of litter is not less than 800 thousand in the presence of small oocysts - 60-70%, medium - 25-35%, large - 5% of the total.

With significant quantitative fluctuations in the categories of oocysts, the final decision is made based on the results of a vaccine immunogenicity test.

Determination of immunogenic activity.

Equipment and materials

For testing use the equipment and materials specified in clause 3.3.

Test

The test is carried out on 5-day-old chickens free of coccidia. The absence of oocysts in fecal matter is determined according to GOST 25383.

Chickens of 5 days of age are divided into two groups (experimental and control) according to the principle of analogues of 10 goals in the group. The birds of the experimental group are injected into the goiter one dose of the vaccine and transferred to the floor. The chickens of the control group are kept in the cage, and they receive a coccidiostatic in a prophylactic dose. After 20 days, the birds of both groups were administered 150 immunizing doses of the vaccine, excluding coccidiostatic from the feed of chickens in the control group three days before infection. Observation is carried out for 10 days.

A vaccine is considered immunogenic if among the chickens of the experimental group there is no mortality from coccidiosis (a slight presence of blood in the litter is allowed) with a mortality from coccidiosis of at least 20% among chickens in the control non-immune group.

In the case of death of birds from coccidiosis in the experimental group, the experiment is repeated on twice the number of vaccine samples.

If repeated negative results are obtained, the vaccine series is rejected.

3.5. Sterility determination

The determination of sterility is carried out in accordance with (9).

The vaccine must be sterile against bacterial and fungal flora.

4. Storage conditions and shelf life of the vaccine.

4.1. The vaccine is stored in a dark place at a temperature of + 2-6 ° C.

4.2. The shelf life of the vaccine is 6 months from the date of manufacture.

The essence and practical applicability of the claimed invention is illustrated by the following examples.

Example 1

The results of a study of the effectiveness of a four-type vaccine in experimental conditions

Table 1 shows the data obtained in the experiment on chickens, which were given a 4-type vaccine at 5 days of age, and after 20 days control infection of 4 coccidia species with live oocysts at a dose of LD _{50 was} carried out. The control was chickens unvaccinated, infected at the age of 25 days, by live oocysts of 4 species of coccidia in a dose of LD ₅₀ , as well as unvaccinated uninfected (the so-called “clean” control).

Table 1. The results of a study of the effectiveness of a four-type vaccine in experimental conditions Group Assignment The number of goals in the group Vaccination at 5 days of age Control infection at 25 days of age The percentage of death of chickens from coccidiosis The percentage of surviving chickens Experienced vaccinated 10 + + 0 one hundred Control unvaccinated infected 10 - + fifty fifty Control unvaccinated uninfected 10 - - 0 one hundred

The results showed that a 4-type vaccine protects 100% of chickens from infection with coccidiosis, while only 50% of chickens survived from unvaccinated vaccines. Thus, it was found that the 4-species vaccine is immunogenic and can, along with CC, be used for specific prophylaxis of coccidiosis.

Example 2. In order to increase the shelf life of a 4-type vaccine against coccidiosis to replace formaldehyde, which is used as a preservative (disinfectant) in the manufacture of CC, the activity of chloramine and ecocide (vircon) was studied in laboratory experiments.

Table 2. The effect of various preservatives (disinfectants) on the shelf life of a 4-type vaccine №№ Group Assignment Disinfectant Shelf life, months Number of goals Control infection result Palo (goals) Survival rate one Experienced Formalin 2% 3 10 0 one hundred 2 Experienced Formalin 2% 5 10 one 90 3 Experienced Formalin 2% 6 10 2 80 four Experienced Chloramine 1% 3 10 0 one hundred 5 Experienced Chloramine 1% 5 10 0 one hundred 6 Experienced Chloramine 1% 6 10 0 one hundred 7 Experienced Ecocide 2% 3 10 0 one hundred 8 Experienced Ecocide 2% 5 10 0 one hundred 9 Experienced Ecocide 2% 6 10 0 one hundred 10 Experienced - - 10 0 one hundred eleven Control infected - - 10 four 60 12 Control uninfected - - 10 0 one hundred

The data obtained as a result of the experiment show that in groups of chickens vaccinated with oocysts treated with chloramine and ecocide (Nos. 4, 5, 6 and 7, 8, 9, respectively), the survival rate of birds after infection after 5 and 6 months of storage was 100 %, while in the groups treated with formalin (Nos. 1, 2, 3) 90 and 80% survived, respectively. Thus, replacing the disinfectant in the vaccine with chloramine or ecocide can increase its shelf life to 6 months.

Example 3. Table 3 shows the data obtained in the experiment on chickens who were used 4-type vaccine with food and method of feeding.

Table 3. The effect of the method of using the vaccine on its effectiveness Group Assignment Vaccination at 5 days of age Control infection at 50 days of age The percentage of death of chickens from coccidiosis Survival rate Experienced (with food) + + - one hundred Experienced (soldering) + + - one hundred Control (unvaccinated) - + fifty fifty Control (uninfected) - - ^- one hundred

It was shown that vaccination of chickens against coccidiosis by the method of feeding by water is as effective as the use of a vaccine with food. Moreover, the method of watering is less time-consuming and allows you to achieve a more uniform production of immunity in the entire livestock.

Example. Table 4 shows the immunization data for chickens with different doses of vaccine.

Table 4. Immunization results for chickens with different doses of vaccine. No. gr. Immunizing dose Age per day immunize. 1st immunization. Number of goals Dose KZ Control infection results Fell Observation and autopsy findings 1 immunization. 2 immunize. one 0.5 ID 0.5ID 5 10 150 ID 0 At 5 cps, the chickens are slightly depressed, the litter is liquid, with bloody streaks 2 1.0 ID 1.0 ID 5 10 150 ID 0 All chickens are healthy for 10 days after SC, litter is normal 3 1.5 ID 1.5 ID 5 10 150 ID 0 All chickens are healthy for 10 days after SC, litter is normal four No No No 10 150 ID four Severe oppression, refusal of food, liquid foamy droppings with streaks and pieces of mucous membrane, opening of dead chickens revealed hemorrhagic lesions of the middle part of the small intestine and blind processes, characteristic of coccidiosis

The content of the chickens is cellular, the first immunization is at 5 days of age, the second is 7 days after the first.

One immunizing dose (ID) contains: E.acervulina - 3000, E.maxima - 500, E.necatrix - 1000, E.tenella - 1000 sporulated oocysts.

Control infection (KZ) was performed 14 days after the second immunization at a dose of 150 ID.

Chickens of the 4th (control) group were kept in pre-short-term conditions under conditions excluding their spontaneous infection with coccidia. Observation of chickens of all groups was carried out for 10 days after short-circuit.

Thus, the claimed vaccine for the immunization of coccidiosis (eimeriosis) of chickens has high immunogenic properties, extended shelf life, is environmentally friendly and economical, more technological to use.

Information sources

1. Edgar S.A. Coccidiosis of chickens and turkeys and control by immunization. Proc. 11 th World Poultry Congress. Mexico City, 1958.

2. Williams R.B. Progress towards anticoccidial vaccines for broiler chickens / Shering-Plow Animal Health, 2000.

3. Immunoprophylaxis of coccidiosis in birds. Symposia biologica hungarica, 33, 1986.

4. Ananiev N.B., A.I. Kirillov, Krylova N.P. Immunochemical prophylaxis of eimeriosis of chickens. Veterinary Medicine, No. 6, 1987.

5. RU, application No. 2009129306/15 A, C12N 1/20, A61K 39/012.

6. RU, application No. 2009129341/15 A, C12N 1/20, A61K 39/012.

7. RU, application No. 2009129305/15 A, C12N 1/20, A61K 39/012.

8. RU, patent No. 2312890.

9. GOST 28085-89. Biological preparations. The method of bacteriological control of sterility. Moscow, Standardinform, 2007.

Claims (1)

A vaccine for the prevention of coccidiosis (eimeriosis) of chickens containing a suspension of a mixture of coccidia oocysts and a preservative, characterized in that it contains a suspension of a mixture of sporulated oocysts of strains of the causative agent of coccidiosis deposited in the collection of GNU VNIVIP of the Russian Agricultural Academy -
a strain of the causative agent of coccidiosis Eimeria acervulina L-2-15 KE No. 2,
strain of the causative agent of coccidiosis Eimeria maxima L-3-2 KE No. 3,
a strain of the causative agent of coccidiosis Eimeria necatrix L-4-18 KE No. 4,
a strain of the causative agent of coccidiosis Eimeria tenella L-1-23 KE No. 1 in a ratio of 3000: 500: 1000: 1000 sporulated oocysts in a single immunizing dose, and as a preservative, a 1% aqueous solution of chloramine.