The bacterial strain pseudomonas aeruginosa 5211used for the manufacture of a vaccine against pseudomonas fur-bearing animals

 

(57) Abstract:

The invention is intended for the production of vaccines against Pseudomonas aeruginosa infection fur-bearing animals, mainly foxes and polar foxes. The bacterial strain Pseudomonas aeruginosa 5211deposited in the collection of strains of pathogens fur-bearing animals of the Museum of bacterial and viral cultures research Institute fur farming and breeding (NIPSC). C. A. Afanasyeva and has a registration number 5211. Strain refers to 06 serovariants. The strain is cultivated in broth Martin or synthetic medium at 32oC for 18 - 20 h with constant shaking. After controlling for grown culture on toxigenicity and proteolytic activity separated backass subjected to inactivation. Next add the adjuvant is aluminium hydroxide and Packed. The invention allows to prevent hidden losses during the reproduction of fur-bearing animals in farms prone to Pseudomonas aeruginosa infection, regardless of the serological profile of the pathogen.

The invention relates to the field of biotechnology and veterinary medicine and is intended for the production of vaccines against Pseudomonas aeruginosa, infectio, causes significant economic damage to the farming and poses a risk to human health. At animal registered each year, anoothi among Minks, foxes and polar foxes. Registered cases of the disease in rabbits, otters, ferrets. In recent years, began to celebrate the Pseudomonas infection in dogs home detention.

Pathology during Pseudomonas aeruginosa infection in people, so the animals are very diverse - damage to the reproductive system, septic processes in the body, the gastro-intestinal tract in young animals, lesions of the mammary glands, etc. foxes and Arctic foxes it flows with genital lesions and is accompanied by disruption of pregnancy, abortion, resorption of embryos, proputovanju females.

The causative agent of Pseudomonas has Serafimovo variability, therefore, the effectiveness of specific prophylaxis is determined by the completeness of the antigenic composition of the vaccine components.

Known vaccinal strain of Pseudomonas aeruginosa PA-7 second serotype used to produce a toxoid vaccine against Pseudomonas aeruginosa infection (SU N 1202109, publ. 15.06.86).

However, the known strain does not contain the antigens of the pathogen Pseudomonas push is the closest analogue is the strain Pseudomonas aeruginosa N202 sixth serotype, included in the inactivated vaccine Pseudomonas fur-bearing animals, mainly mink ("Veterinary drugs". The Handbook, edited by D. F., Osize., M.: Kolos, 1981, S. 306-308).

However, the known production strain is unstable virulence and immunogenic properties and, moreover, does not prevent Pseudomonas foxes and foxes caused by pathogens other serovariants.

The objective of the invention is to obtain a strain of Pseudomonas aeruginosa for the production of an effective vaccine against Pseudomonas fur-bearing animals, regardless of the serological profile of the pathogen.

The technical result of the invention is to obtain a new strain Psendomonas aeruginosa 5211able to produce the main pathogenicity factors in the optimal values and help to produce a vaccine against any serovariants Pseudomonas aeruginosa not only for foxes and polar foxes, and other susceptible to this infection animals.

The inventive bacterial strain Pseudomonas aeruginosa 5211selected from female Fox C/s "Timkovsky" and deposited in the collection of strains of pathogens diseases of fur-bearing animals of the Museum of bacterial and viral coltee registration number 5211.

Strain Ps. aeruginosa 5211intended as a producer of proteolytic enzymes (alkaline protease and elastase) in the manufacture of specific immune drug against Pseudomonas aeruginosa infection of foxes and polar foxes.

The bacterial strain Pseudomonas aeruginosa NYPSC 521 N1characterized by the following features and properties.

Genetic markers.

Serovariants 06 (Habs)

Piacenti 268678

Strain 5211proteolytic.

Morphological properties. Workplace culture is a thin straight gram-negative bacilli, size 2-3 x 0.5 micron. Sticks moving, monotachi, do not form spores, smears are arranged singly or in rows.

Cultural properties. Aerobe, optimum temperature for growth at 32-37oC retains the ability to grow and reproduce under 42oC. Grows in almost all nutrient media with a pH of 8.0. On the surface BCH forms a characteristic grayish-silvery film. Forms a pigment pyocyanin. The best medium to cultivate an environment Iensen'a, enriched with salts and aminopeptidase. On the IPA forms S - and R-forms, with a predominance of S-forms.

Biochemical activity. High chislom gluconic acid without gas, liquefies gelatin, roll milk does not cause formation of indole and hydrogen sulfide. Forms the cytochrome oxidase, causes the hydrolysis of ndimethylacetamide.

Virulent properties. Intraperitoneal injection white mice LD/50 is 1.3 107CFU/ml.

Antigenic properties. The strain produces alkaline protease, exotoxin A.

Toxigenic properties. In a liquid nutrient medium produces alkaline protease (figure hydrolysis of casein 320), exotoxin A (TPHA 1:128).

The deposited strain. At a temperature of 62oC in vials in a dried state for more than three years; in test tubes in a column semisolid agar cooked in broth Martin with the addition of 0.5% of sodium glutamate and 1% of glycerol, vaseline under the oil for three months; in test tubes on the sloped agar no more than 14 days.

The control strain. Control of morphological, biochemical, cultural and virulent properties at least 1 times in three months. Antigenic properties (protease and elastase) check before production of immune drug. The pre-strain is cultivated in an environment Iensen'a.

The rate of hydrolysis of casein alkaline protease in the reaction SIAs.

Example 1. The bacterial strain Pseudomonas aeruginosa 5211cultured broth Martin or synthetic medium at 32oC for 18-20 hours with constant shaking at shuttel machine. After a period of incubation, the culture is checked for purity growth in smears stained by Gram.

Forth from the grown culture selected sample, centrifuged it at 6000 rpm. /min for 20 minutes the Supernatant is drained and examined the contents: exotoxin And reactions precipitation (RP) with antitoxic serum and/or alkaline protease in the reaction of hydrolysis of casein, and/or elastase in RP with monovalent antielastase serum. The title should be: exotoxin A - not less than 1:4, alkaline protease - not less than 1:32, elastase is not less than 1:16.

After controlling for grown culture on toxigenicity and proteolytic activity separated backass subjected to inactivation by any known method and add an adjuvant such as aluminum hydroxide.

Example 2. Obtained according to example 1, the vaccine is tested for safety on the 9th too. The vaccine is injected intramuscularly in the thigh right paw in doses: three Arctic foxes on 1.0 ml, three 2.0-ml three 3.0 ml.

Conduct daily nabt twice on the 2nd and 4th days after drug administration.

The results showed that all Arctic foxes behaved normally, ate food completely, aggression was not observed. Body temperature was within the normal range 37-39oC. by touching the point of injection of the vaccine was not detected limb swelling, pain.

Example 3. Received vaccine subjected to immunization foxes on the farm prone to Pseudomonas aeruginosa infection, to study the effect of immunization on the results of reproduction, since the primary pathology in Pseudomonas infection is manifested in the period of reproduction.

In this economy among foxes in some cases abortion, proponowania females, the birth of dead puppies. However, aborted females in the last stages of pregnancy (the week before the deadline of security) bacteriological examination was allocated to culture Pseudomonas aeruginosa 03 and 09 serovars.

Obtained in example 1 subjected to immunization vaccine 50 females foxes intramuscularly in the dose of 1.5 ml quality control are the other 50 females of similar age, color, and conditions and feeding.

It is established that the vaccine increased the fecundity of Arctic foxes experimental group 0.7 puppy. The vaccine prevent the s was registered litter just 4-5 puppies. This is most often caused by resorption of fetuses in the first half of pregnancy.

Thus, a vaccine based on strain 5211to prevent hidden losses during the reproduction of fur-bearing animals in farms, troubled by Pseudomonas aeruginosa infection.

The bacterial strain Pseudomonas aeruginosa 5211used for the manufacture of a vaccine against Pseudomonas fur-bearing animals.

 

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5 cl, 8 dwg, 10 ex

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