SU1018971A1 - Method for preparing culture medium for growing fluorobacteria photobacterium leog nathi - Google Patents

Abstract

THE METHOD OF PREPARING THE FOOD MEDIUM FOR THE DISSEMINATION OF PHOTOBACTERIA Photobactorishn leognathi ,. involving the preparation of an initial nutrient medium containing carbon dioxide nitrogen, phosphorus and mineral salts, followed by the growth of photobacteria on it, so that, in order to improve the quality of the medium by accumulating in it the luminescence intensity of the photobacteria, the original nutrient medium is mixed in a ratio of 2: 3 with sterilized spent culture liquid, obtained by growing the photobacteria on the initial nutrient § for 2-3 hours and after the onset of (L of luminescence intensity and subsequent separation of bacterial C "cells.

Description

00

t.

Yu

i “i i h

Fig /

The invention relates to microbiology, in particular to methods of cultivating luminous bacteria, and can be used to create indicator systems.

There is a method of growing photobacteria on a nutrient medium containing sources of carbon, nitrogen and mineral,

However, the use of such an environment does not provide a high luminescence intensity.

Closest to the invention, by technical nature and the effect achieved, is a method of preparing a nutrient medium for growing photobacteria Photobacteriiuni leognathl. , provide for the preparation of the original nutrient medium containing sources of carbon, nitrogen, phosphorus and mineral salts, with the subsequent cultivation of photobacteria on it,

According to this method is prepared

nutrient medium of the next.

/ tava, g: saC1 30 KN2ROD1 NdO 0, .04-i2 Н, 0 6; (NH)., 5j citrate Na-0.65; peptone 5; glycerin3 ml per 1 liter of distilled water G23.

The disadvantage of the known method is the low maximum specific intensity of photobacteria luminescence. (10 μA) due to the low content in the original nutrient medium of the out-inductor of luciferase synthesis.

Goal: The invention is an improvement in the quality of the media due to accumulations in substances that show the intensity of the photobacteria.

The goal is achieved by the fact that, according to the method of preparing the nutrient medium for extinction -... in this case, the original nutrient medium is mixed in a 2: 3 ratio with a pre-sterilized spent culture fluid obtained by growing the photobacteria on the original nutrient medium for 2-3 hours after the onset of age. Neither the intensity of luminescence and the subsequent separation of bacterial cells.

The essence of the method lies in the trace.

On the original semi-synthetic medium, bacteria Photobacteriiu are grown. leognathi strain 54. The cultivation process is carried out with aeration at 28-30 sec. In the phase of increasing luminescence intensity, separate the cultured medium from bacteria, sterilize it and add liquid to it.

which semi-synthetic medium at the rate of: for 3 hours of processed medium 2 hours semi-synthetic. Fresh photobacteria are seeded in the semi-combined medium and cultured with aeration at 28-30 ° C.

FIG. Figure 1 shows that the maximum specific intensity of luminescence on a combination medium (curve 1) is 500 µA. FIG. 2 - dynamics of specific intensity of luminescence in semi-synthetic medium under standard conditions of cultivation (known method).

Sowing into both media was carried out from the same cell suspension, at the same time and in the same dose.

Example 1. Prepare a liquid polysynthetic medium of the following composition, g: NaCl 30. KH-jPO l.fMgSQ.7НгО 0.2; Na, HPO; j-12HiO 0.6 / (NH), 5, .at Na 0.65, peptone 5, glycerin - 3 MJ per 1 l of distally. bathroom water. For the solid erie, more agar-agar is added at the rate of 22 g / l.

Bacteria grown on solid semi-synthetic medium are suspended in a semi-synthetic liquid medium until a homogeneous initial suspension is obtained. In a flask with a volume of 250 ml, containing 50 ml of semi-synthetic liquid medium, seeded with 1 ml of feed suspension. Cultivation is carried out on a rocking chair at 28–30 C. The density of the baterium is taken from C 1 1 photocolorimetric. on FEK-54 (blue filter). The luminescence is measured on a setup for recording the luminescence. In the phase of increase in luminescence intensity (usually after 2-3 hours from the start of cultivation), bactercal cells are precipitated by centrifugation at 7000 rpm for 20 minutes. The treated culture medium is sterilized at 0.5 atm, and it can be stored for use in combination media. The combined medium is prepared at the rate of: 3 hours by processing the medium and 2 hours by the liquid semi-seynt.e. medium.

Secondary culturing is carried out in 250 ml flasks containing 50 ml of the combined medium (30 ml of additionally + 20 ml of the usual semi-synthetic). The original seed suspension is prepared in the usual way, i.e. Bacteria grown on solid semi-synthetic medium are suspended in semi-synthetic semi-synthetic medium until a homogeneous suspension is obtained.

According to the I ml of this suspension, they are seeded in flasks with a combined medium, cultivation is carried out on a quality basis, at 28–30 ° C with aeration. After 8–10 h after the start of cultivation, the specific intensity of luminescence reaches a maximum.

Claims (1)

METHOD FOR PREPARING A NUTRIENT ENVIRONMENT FOR GROWING A PHOTOBAKT 'RIY Photobacterium leognathi ,. involving the preparation of the original ^! a nutrient medium containing sources of carbon, nitrogen, phosphorus and mineral salts, followed by the cultivation of photobacteria on it, which can be done in order to improve the quality of the environment due to the accumulation of substances in it, which increase the luminescence intensity of photobacteria, the initial nutrient medium is mixed in a 2: 3 ratio with sterilized spent culture fluid obtained from growing photobacteria on the initial nutrient medium for 2–3 h and after the onset of increase in luminescence intensity and the following tdelenii bacterial 'cells. · Fig / in C> 1 ¹ 1018971