SU1247419A1 - Method for reversion of tuberculosis l-form microbacteria - Google Patents

Description

eleven

The invention relates to veterinary medicine, in particular, to the study of the variability of the pathogen of the tuberculosis pathogen.

The purpose of the invention is to accelerate the method.

The method is carried out as follows.

7-day-old chicken embryos are taken, the location of the embryo is determined by ovoscoping. Then, following the rules of asepsis, the embryos in the yolk sac are infected by cultures of L-forms of mycobacterium tuberculosis. The Pasteur pipette collects the culture of the L-forms from the modified medium with the Shkol'nikovoi and refills the sterile syringe. After that, the shell of the embryo with air over the air space is treated with alcohol, a hole is pierced in its center, and infection of 0.1-0.2 ml is produced in the yolk sac, located on the opposite side of the embryo. Then the hole in the shell is filled with paraffin and the infected embryo is placed in a thermostat with a temperature of 35-37 C and humidity of 60-65%. After one day, the hole above the air chamber is opened,. the Pasteur pipette collects part of the yolk for preparing smears and coloring them according to Zil-Nielsen, as well as seeding on nutrient mediums of Levenstein-Jensen. The same yolk is infected with fresh 7 dpevp embryos, in this dose. And so in the last 5 before receiving the reversion that occurs, starting with the 3-4th passage. It is found in painted smears where acid-resistant sticks are detected. The growth of typical pathogenic colonies occurs on the 2-14th day.

dV

depending on the type of mycobacterium tuberculosis.

Example 1. A culture of L-forms of mycobacteria grown on Shkolnikova’s semi-liquid medium in a modification of I. R. Dorozhkova} was grown from a material from animals. During phase contrast microscopy of native preparations, microstructural elements of L-forms were found in the form of clusters of

VNIIPI Order 4080/26 Circulation 490 Subscription Proizv.-polyr, pr-tie, Uzhgorod, st. Project, 4

5 0 5 about 0

5 Q

five

vacuolation elements, spherical TEL c of uniform ground mass, s; In order to obtain a yolk sac reversal of 7-day-old chick embryos, O, -O, 2 ml L-culture was introduced. Infection and passage are carried out according to the described method. A day later, a preparation was prepared from the yolk of an infected embryo and stained using the Zil-Nielsen method. Microscopy of the drug shows the presence of clusters of spherical vacuolization of elements and granular masses, painted in blue. Microscopic examination of a smear, an embryo made from the yolk of a passage, shows non-acid-resistant n; 1 blocks of various sizes and shapes. There are also clusters of spherical elements. Microscopy of a smear from the embryo of the 3rd passage determines the presence of separate acid-resistant rods in a mixture with non-acid-resistant, In the preparations of the passage, acid resistance is well expressed. From the yolk of this embryo, it is sown on Levenshteyn-Jensen medium, and after 14 days, growth of typical mycobacterial colonies is observed.

Example 2 A culture of L-forms of mycobacteria, isolated from cattle lymph nodes, infected with 7-day-old chicken, embryo, B colored preparations of the first passage look at acid-resistant granular mass and clusters of spherical bodies, painted in blue dvet. Microscopy of the preparation from the 2-HS embryo passage along with an acid-cleaned granular mass revealed non-acid-resistant filaments. At the 3rd passage, acid-resistant Rods and Yarns mixed with non-acid-resistant materials were found. At the 4th passage, the presence of acid-resistant bacteria is established. When sowing yolk from the embryo of the 4th passage on the 20th day, the growth of typical mycobacterial colonies was obtained

Pa raspid environments

R

The use of the invention makes it possible to speed up the time needed for obtaining revertants of L-forms of mycobalsterium tuberculosis.

Claims (1)

METHOD FOR REVERSING L-FORMS OF TUBERCULOSIS MYCOBACTERIA, including multiple passaging of a suspension of L-forms of mycobacteria through a living organism with simultaneous monitoring by culture on solid nutrient media and smear microscopy, characterized in that, in order to accelerate the method, passaging is carried out with an interval of one: a day through 7-day chicken embryos, while a suspension of L-forms of mycobacteria is used at a dose of 0.1-0.2 ml per chicken embryo. I 2474 I 9