Way of intestine dysbacteriosis prevention during experiment

FIELD: medicine.

SUBSTANCE: invention concerns experimental medicine and can be used in veterinary science. The Ultrasorb preparation is fed to the animals that are on a vivarium standard diet, within 14 days, based on 3 g/day, admixed with forage. The way prevents development of intestinal dysbacteriosis at animals.

EFFECT: prevention of intestinal dysbacteriosis at animals.

3 ex, 2 tbl

 

The invention relates to experimental medicine and can be used for prophylaxis of a dysbacteriosis of the intestine in experimental animals (white rats) during the experiment and during the animals in the experimental biological clinics (vivaria).

In the available scientific and patent literature methods of prophylaxis of a dysbacteriosis of the intestine in vivo was not found.

The invention is directed to solution of the problem: the creation of a method of prophylaxis of a dysbacteriosis of the intestine in experimental animals resulting from improper feeding of animals.

Despite the relative stability of the intestinal microflora of rats, there may be violations under the influence of various, often exogenous factors: violations of food, replace in the diet of some other products, lack of vitamins in the diet. The existing imbalance of the intestinal microflora has a negative impact on the results of the experiment.

Effect: preventing the development of dysbiosis in experimental animals with the aim of developing a method of prophylaxis of intestinal dysbacteriosis in humans.

This task is achieved by prophylactic use in experimental animals (white rats) for 14 d is her drug "Ultrasorb" at the rate of 3 g/day, mixing it with food. Biologically active additive "Ultrasorb" issued by CJSC "Berry", Kirov, D. Ugrina. This product is used as a preventive measure practically healthy people when exposed to adverse environmental and production factors, with the increased risk of the formation of goiter, cancer, diabetes, immune deficiency. It is also used alone or as a component of functional food in the gastro-intestinal tract (stomatitis, dysbiosis, acute intestinal infections, ulcer of stomach and duodenum, gastritis, cholecystitis, hepatitis, enteritis, colitis), metabolic disorders (atherosclerosis, obesity, diabetes, hyperglycemia), alcohol intoxication and alcoholic pathology in inflammatory pathologies of the skin from bacterial and allergic origin in the complex therapy of secondary immune deficiency. The main component of "Ultrasorb are fermented insoluble and soluble dietary fiber, which helps to normalize the intestinal microflora.

The method is as follows. The drug "Ultrasorb" after mechanical grinding in a mortar and added to feed for experimental animals (white lab the lab rats) in an amount of 0.1 (10%) of the daily volume of food (30 g/day per animal according to the existing norms) - 3 g/day. Prophylactic use of the drug "Ultrasorb" is 14 days.

The experiment was performed on 10 animals male and female outbred white rats weighing 168,226,5 g, four months of age contained in standard vivarium conditions. All experiments were conducted in accordance with "the Rules of work with the use of experimental animals" (Annex to order of the Ministry of health of the USSR from 12.08.1977, N 755) and the European Convention for the protection of vertebrate animals used for experimental and other scientific purposes" dated 18 March 1986 Until the beginning of the experiment the animals were on a normal diet vivarium in accordance with the rules approved by order of the Minister of health of the USSR from March 10, 1966, No. 163 and the Order of the Ministry of health of the USSR from 10.10.83 No. 1179 (paragraph 4.1):

- Feed-concentrate granulated 50.0 g;

- Low fat cottage cheese - 6.0 g;

- Carrots - 20,0 g;

- Greens of sprouted oats - 15.0 g;

- Total feed, 91,0.

After registration of the original indicators in the diet of animals as a component of functional foods was included "Ultrasorb". Course villas animals "Ultrasorb amounted to 14 days.

During the experiment twice, at the beginning of the experiment and after 14 days was carried out by examination of feces on a dysbacteriosis.

Primary sowing times is the origin of faeces was performed on differential diagnostic environment: bifidobacteria - on Wednesday Blaurock, lactobacilli - on environment, MPC-2, enterobacteria - on environment Endo, Staphylococcus - yolk-salt agar, enterococci - milk-salt agar with polymyxin, fungi - on environment Saburo, with subsequent characterization of selected bacteria by standard methods. Similar to the spectrum of the analyzed microorganisms due to the fact that when dysbiotic conditions in the intestinal biocenosis primarily changes the content of these microorganisms. The number of organisms expressed in decimal logarithm (lg) colony forming units (CFU) per 1 g wet weight of faeces.

The experimental results were subjected to statistical analysis using the software package patch BIOSTAT. To calculate the arithmetic mean of the standard deviation. The reliability of the obtained differences were determined using the criterion of Wilcoxon signed. When determining background values obtained data were compared with the average population fluctuations. When determining after 14 days after inclusion in the diet of animals "Ultrasorb" - with the original figures.

The animals in the beginning of the study in the intestine was determined by the following microorganisms: bifidobacteria, lactobacilli, enterobacteria, lactose-negative enterobacteria, enterococci, staphylococci, fungi, Bacillus.

Table. 1
Quantitative characterization of microorganisms living in the habitat of the large intestine of rats before the start of the experiment, lg CFU per 1 g wet weight of faeces (MM).
MicroorganismsExperienced groupThe limits of fluctuations of population values for animals of a given species*
Bifidobacteria (MM)6,41,88,0-9,0
Lactobacillus (MM)7,21,7of 8.3 to 9.6
Enterobacteriacea (MM)5,60,55,3-6,4
Enterobacteriacea (lactose-negative) (MM)5,60,5**
Enterococcus spp (Mm)8,0**
Staphylococcus spp (Mm)5,80,84,5-7,1
Candida spp (MM)1,81,1**
Saccharomyces spp (MM)2,41,5**
Bacillus spp (MM)2,41,5**
* data HE batishcheva et al. The nutrition. - 2002. No. 3. - T. - P.36-39.
** data characterizing the limits of fluctuations of population sizes of organisms in the contents of the large intestine of rats, were not found in literature

The analysis of the content of protective microflora (bifidobacteria and lactobacilli) showed that at the beginning of the experiment, the number of bifidobacteria and lactobacilli in the biotope of the colon of rats was somewhat lower average population size (table 1). Before the start of the experiment the content of microorganisms, belonging to the family Enterobacteriacea, in the habitat of the large intestine of experimental animals was 5.60,5 lg CFU/g with an average population size of 5.3 and 6.4 lg CFU/g While the content of lactose-negative enterobacteria was high: their concentration was comparable to the concentration of all enterobacteria and 5.60,5 lg CFU/g, that is, in the composition of enterobacteria noted the apparent prevalence of lactose-negative forms. As is known, increasing the population of lactose-negative enterobacteria is an indicator of dysbiotic changes. Additionally, intestinal biocenosis of rats was determined by fungi, bacteria, yeast

Table. 2
Quantitative characterization of microorganisms living in the habitat of the large intestine of rats after 14 days from the beginning of the experiment, lg CFU per 1 g wet weight of faeces (MM).
The microorganismThe presence of bacteria in the feces of animals, CFU/g, lg Source dataThe presence of bacteria in the feces of animals, CFU/g, lg After 14 from the beginning of the experiment
Bifidobacteria (MM)6,41,86,81,1
Lactobacillus (MM)7,21,78,20,5
Enterobacteriacea (MM)5,60,55,01,0
Enterobacteriacea (lactose-negative) (MM)5,6+0,50*
Enterococcus spp (Mmm)8,08,0
Staphylococcus spp (MM)5,80,85,20,5
Candida spp (MM)1,81,1
Saccharomyces spp (MM)2,41,50*
Bacillus spp (MM)2,41,52,01,2
* - p<0.05 with respect to baseline indicators

As can be seen from table 2, after 14 days after inclusion in the diet of animals "Ultrasorb" the content of lactobacilli in the biotope of the colon of rats reached the average population size and amounted to 8.20,5 lg CFU/g Concentration of bifidobacteria during the observation period also tended to increase, though less pronounced.

After 14 days of giving animals "Ultrasorb" there was a decrease in the concentration of Enterobacteriaceae (5,01,0 lg CFU/g) and the disappearance of lactose-negative enterobacteria. Thus, under the influence of "Ultrasorb has been a change in the population of enterobacteria in the normalization of the indices.

Animals treated "Ultrasorb" as a component of functional foods, there has also been a decrease in the concentration of bacteria belonging to the genus Staphylococcus. So, in the beginning of the experiment the concentration of staphylococci in the colon of rats was 5.80,8 CFU/g at the rate of 4.5 to 7.1 CFU/g, after giving "Ultrasorb" - 5,20,5 CFU/g, the course of reception "Ultrasorb" also contributed to the San is the large intestine of experimental animals from fungi and yeast, the trend towards a decrease in the concentration of bacilli.

Thus, the application of Ultrasorb" as a component of functional foods from animals on a standard diet has a beneficial effect on the normalization of quantitative and qualitative characteristics of intestinal biocenosis, which makes possible its use for the prevention of development in experimental animals of dysbiosis.

Examples of specific performance.

Example 1. Female white rats, weighing 154 g, within 14 days were on a standard diet of the vivarium, and then in sterile conditions sample was collected faeces, subjected to investigation on a dysbacteriosis. In the composition of intestinal biocenosis of the animal was determined by the following microorganisms: bifidobacteria in concentrations of 104CFU/g lactobacilli - 105CFU/g, enterobacteria - 106CFU/g, including lactose-negative enterobacteria - 106CFU/g Enterococcus - at a concentration of 108CFU/g Staphylococcus - 106CFU/g, yeast - 106CFU/g Bacillus

- 106CFU/g After registration of the original indicators in the diet of the animal for 14 days was included "Ultrasorb"that the animal was fed in a mixture with the feed in an amount of 3 g/day.

After 14 days, feces analysis on disbacteriosis the animal was taken again. In the intestinal the biocoenosis were determined: bifidobacteria - in the amount of 105CFU/g lactobacilli - 108CFU/g, enterobacteria - 105CFU/g of lactose-negative enterobacteria was not determined, enterococci - 108CFU/g Staphylococcus - 105CFU/g fungi and bacilli not found.

Example 2. Male white rats, weighing 180 g for 14 days were on a standard diet of the vivarium, and then in sterile conditions sample was collected faeces, subjected to investigation on a dysbacteriosis. In the composition of intestinal biocenosis of the animal was determined by the following microorganisms: bifidobacteria in concentrations of 107CFU/g lactobacilli - 107CFU/g, enterobacteria - 105CFU/g, including lactose-negative enterobacteria - 105CFU/g Enterococcus - at a concentration of 108CFU/g Staphylococcus - 107CFU/g fungi and bacilli not found. After registration of the original indicators in the diet of the animal for 14 days was included "Ultrasorb"that the animal was fed in a mixture with the feed in an amount of 3 g/day.

After 14 days, feces analysis on disbacteriosis the animal was taken again. In the intestinal biocenosis were determined: bifidobacteria in the amount of 108CFU/g lactobacilli - 108CFU/g, enterobacteria - 106CFU/g of lactose-negative enterobacteria was not determined, enterococci - 108CFU/g Staphylococcus - 105CFU/g fungi and BAM is lly not found.

Example 3. Female white rats, weighing 155 g, within 14 days were on a standard diet of the vivarium, and then in sterile conditions sample was collected faeces, subjected to investigation on a dysbacteriosis. In the composition of intestinal biocenosis of the animal was determined by the following microorganisms: bifidobacteria in concentrations of 107CFU/g lactobacilli - 107CFU/g, enterobacteria - 105CFU/g, including lactose-negative enterobacteria - 105CFU/g Enterococcus - at a concentration of 108CFU/g Staphylococcus - 106CFU/g Bacillus - 106CFU/g fungi were not identified. After registration of the original indicators in the diet of the animal for 14 days was included "Ultrasorb"that the animal was fed in a mixture with the feed in an amount of 3 g/day.

After 14 days, feces analysis on disbacteriosis the animal was taken again. In the intestinal biocenosis were determined: bifidobacteria in the amount of 108CFU/g lactobacilli - 108CFU/g, enterobacteria - 104CFU/g of lactose-negative enterobacteria were not found, enterococci - 108CFU/g Staphylococcus - 106CFU/g Bacillus - 105CFU/g

Method of prevention of dysbiosis in the experiment, which consists in the fact that the experimental animals that are on the regular diet of the vivarium, within 14 days give medication "Ultrasorb" the C at the rate of 3 g/day, mixing it with food.



 

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