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  Отзывы специалистов. Часть 1

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The conclusion about efficiency of developed agents of protection from toxins

версия для печати

The ministry of a defense of Russian Federation. Researchinstitute of microbiology.

The scientific principal of work. The chief of a department of a toxicology.

According to the calendar plan of research activity the efficiency of REKITSEN-RD use was tested in the experiments with intestinal botulinic poisoning. The following materials and methods were used:

  • The substance "REKITSEN-RD", prepared according to Technical Standard 929500- 05344371-001-96 by hard-phase culturing of specially selected Saccharomyces vini strain of on the vegetative substrate No.VKLM U-511; 1,0 gram of a dry product contains (1,0-1,2) x1010 of microbal cells with residual humidity (5,0±1,0) percent.
  • glycerin solution of the Type A botulinic toxin with concentration 108 Units/ml (1 Unit corresponds to 1 LD50 of the toxin injected intraperitoneally to white mice);
  • wildbread white mice of both gender weighting 18...22 g;
  • laboratory balance of VLTK-200 type
  • photoelectrocolorymeter of FEC-56 type
  • thermostat electrical of EU-1125 type
  • centrifuge laboratory of T-23 type
  • tuberculin syringes
  • methylene-blue stain
  • talcum medical.

LD50 parameter was calculated according to the Kerber-Ashmarin formula. ST50 was computed as:

Where ST50 is average time up to destruction
n - number of animals died in time t
N - total of animals taken into experiment.

At the first stage the estimation sorptive capacity of REKITSEN-RD was carried out. The researches were executed according to Federal Standard 4453-74 "Coal activated clarifying wood powdery " with use of a methylene-blue stain. The parameter was determined as 4,2 mg of a stain per 1,0 g of a substance.

Toxin-binding ability of REKITSEN-RD in studies in vitro

Substance
Concentration of a toxin, Units/ml
Amount of the bound toxin, Units/g of a substance
initial
after processing
REKITSEN-RD
1200
420
7800
Talcum
1200
1070
1300

On the final stage the in vivo analysis of REKITSEN-RD toxin neutralizing activity was studied on white mice using a peroral way of introduction.

For this purpose white mice were previously stayed hungry within 24 hours, then the animals were orally entered a portion of type A botulinic toxin and were given a forage at once. The experimental group of animals was taken an admixture of grain and REKITSEN-RD in equal proportion, control group - only grain. The REKITSEN-RD dosage consumed was determined each 24 hours using the rest of a forage and was approximately 1,5-2,0 g/day per 1 mouse. The animals were observed within 4 days, from which per the first 2 days - within 4 hours. By the results of animals' death rate the LD50 value of the toxin for experimental and control groups was determined as well as the ST50 parameter (average time up to death). The results of the series are represented in tab. 2.

Estimation of in vivo toxin neutralizing ability of REKITSEN-RD

Dose of a toxin, УЕ
Share of animals died 1
ST50 value2, hours
LD50 values, Units
REKITSEN-RD
Grain
REKITSEN-RD
Grain
REKITSEN-RD
Grain
3,15х103 0/6 0/6 >96 >96 1,8х104 8,9х103
6,3х103 0/6 2/6 >96 72,86
1,25х104 1/6 4/6 90 52+/-14
2,5х104 4/6 6/6 51+/-8 38+/-11
5,0х104 6/6 6/6 38+/-7 29+/-9
1,0х105 6/6 6/6 29+/-5 17+/-4

The notes.

  1. Numerator - amount of animals died, denominator - number of animals taken in the experiment.
  2. The means of survival time of the animals were given with 95% reliability.
  3. The parameter was calculated with Kerber-Ashmarin method.

Thus the following conclusions are reasonable by results of the work:

  1. REKITSEN-RD is active as a sorptive agent for type A botulinic toxin.
  2. REKITSEN-RD, consumed immediately after botulinic toxin oral introduction, is partially capable to bind and to neutralize the latter with efficiency approximately 3000...5000 Units/g of a sorptive substance, that results in enlarging of animals' average time up to death for 11 ...15 hours.

The offers

To obtain more exact data of REKITSEN-RD sorptive activity the in vivo studies on large experimental animals are needed with oral introduction of the substance in definite amounts

Besides it is seems expedient to study neutralizing ability of REKITSEN-RD in relation to other toxins, for example, shigelligenous, staphylococcal etc.

 

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