The bacterial strain pseudomonas aeruginosa 5132used for the manufacture of a vaccine against pseudomonas fur-bearing animals

 

(57) Abstract:

The invention relates to biotechnology and veterinary medicine and is intended for the production of vaccines against Pseudomonas aeruginosa infection fur-bearing animals, mainly foxes and polar foxes. The bacterial strain Pseudomonas aeruginosa 5132deposited in the collection of strains of pathogens fur-bearing animals of the Museum of bacterial and viral cultures research Institute fur farming and breeding them. C. A. Afanasyeva and has a registration number 5132. Strain refers to 011 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, then add the adjuvant aluminium hydroxide and Packed. The invention allows to prevent the concealment, loss when 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 vaccine prep is under common everywhere, causes significant economic damage to the farming and poses a risk to human health. At animal registered each year, antetype 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 and animals, and is 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 for the manufacture of intoxicatin against Pseudomonas aeruginosa infection (SU N 1202109, publ. 15.06.86).

However, the known strain does not contain the antigens of the pathogen Pseudomonas the

The closest analogue is the strain Pseudomonas aeruginosa N 202 sixth serotype, part of the inactivated vaccine Pseudomonas fur-bearing animals, mainly mink ("Veterinary drugs". The Handbook, edited by D. F., Osize. M.: Kolos, 1981).

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 Pseudomonas aeruginosa 5132able 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 5132selected from female Fox C/s "Plavsk" and deposited in the collection of strains vozbuditelei farming and breeding (NIPSC). C. A. Afanasyeva and has a registration number 5132.

Strain Ps.aeruginosa 5132intended as a producer of exotoxin And in the manufacture of specific immune drug against Pseudomonas aeruginosa infection of foxes and polar foxes.

The bacterial strain Pseudomonas aeruginosa NYPSC 513 N2characterized by the following features and properties.

Genetic markers:

Serovariants 011 (Habs)

Piacenti 887628

Strain 5132toxic forms of the enzyme alkaline protease, slizeobrazujushchej has adhesive activity.

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 environment for the cultivation broth is Martin with the addition of 0.5% of sodium glutamate and 1% glycerol. On MPA activity towards glucose and galactose; in aerobic conditions breaks down sugars to form 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 4,56 105CFU/ml.

Antigenic properties. The strain produces exotoxin A, alkaline protease, extracellular mucilage has adhesive ability against rabbit erythrocytes.

Toxigenic properties. In a liquid nutrient medium produces exotoxin A (titer in TPHA 1:64, RP 1:2), alkaline protease (figure hydrolysis of casein 144), extracellular mucus in the BCH and MPA. The caption adesina against rabbit erythrocytes is 1:32.

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 morphologically, biochemically the Ute before production of immune product.

The titer of exotoxin And TPHA using erythrocytes sensitized immunoglobulin selected from monovalent antitoxic serum, should be not less than 1:64, or in the precipitation reaction with a monovalent antitoxic serum obtained to vysokoochishchennogo the drug exotoxin A, not less than 1:2.

The invention is illustrated by the following examples.

Example 1. The bacterial strain Pseudomonas aeruginosa 5132cultured 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 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 is not below 1:32, elastase is not less than 1:16.

After controlling for grown culture on toxigenicity and about adjuvant, for example aluminium 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 and three - 3.0 mil

Conduct daily monitoring of Arctic foxes within 7 days individually by palpation of the injection site. Thermometry foxes performed 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 normal limits - 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 CLASS="ptx2">

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 prevented the resorption of fetuses in some females of the experimental group, while some females of the control group 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 5132to prevent hidden losses during the reproduction of fur-bearing animals in farms, troubled by Pseudomonas aeruginosa infection.

The bacterial strain Pseudomonas aeruginosa 5132used for the manufacture of a vaccine against Pseudomonas fur-bearing animals.

 

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FIELD: biotechnology, microbiology, medicine.

SUBSTANCE: invention relates to the strain Lactobacillus paracasei CNCM I-2116 used for diarrhea prophylaxis causing by pathogenic microorganisms. Supernatant of this strain culture elicits ability to prevent colonization of intestine with pathogenic microorganisms causing diarrhea also and this strain is designated for preparing agent used for prophylaxis and/or treatment of disorders associated with diarrhea. Agent for oral administration represents therapeutically effective dose of the strain L. paracasei CNCM I-2116 or supernatant of its culture and acceptable foodstuff. Invention provides the enhanced viability of the strain in its applying and effectiveness in prophylaxis of adhesion to intestine cells and invasion to intestine cells of pathogenic microorganisms causing diarrhea.

EFFECT: valuable medicinal properties of strain.

5 cl, 8 dwg, 10 ex

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