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Petri dishes with grown colonies and statistical parameters
of the colonies after 23 hours of experiments are presented on Fig
2: (a) non-activated nutrient medium (control); (b) nutrient medium
activated for 30 minutes; (c) nutrient medium activated for 60 minutes.
Petri dishes after 29 hours of experiment are shown on Fig 3. The
significant inhibition of growth of E.coli in activated samples
was revealed and it confirmed the strong bacteriostatic effect of
MRET-activated medium.
This experiment shows that MRET-activation process has very strong bacteriostatic effect on conditionally pathogenic E.coli microorganisms and that the inhibition of E.coli growth is more effective when activation time is increased. It was observed that at low initial concentration of cells of E.coli in nutrient medium MRET-activation during 30 minutes and 60 minutes period of time inhibited the culture growth NC/N0.5 = 27 and NC/N1.0 = 303 times respectively after 25 hours of experiment (Fig 4). Consequently, the level of bacteriostatic activity was 96% in 30 minutes activated nutrient medium and 99.7% in 60 minutes activated medium. Thus, the direct correlation between bacteriostatic activity of MRET-activated nutrient medium and the time of activation was confirmed. Inhibition of E.coli bacteria growth in aerobic environment
This experiment also revealed the strong effect of MRET-activated water on the process of division of E.coli microorganisms, the modification of forms of culture cells and the size of colonies. It was observed that one of the reasons of abnormally low growth of E.coli bacteria was related to the modification of the process of cell division in MRET-activated nutrient medium. In the process of growth and reproduction a large number of cells did not separate from each other and the linear line-ups consisting of 2-3 sequentially paired cells were formed. The culture cells grown in non-activated and MRET activated medium are shown on Fig 5 and Fig 6 respectively.
Reductant activity is an integral characteristic of metabolic activity of microorganisms and it is measured with the help of Sodium Resasurine color indicator in the percentage degree of discoloration (purple = 0%, red = 50%, transparent = 100%). The reductant activity of E.coli bacteria reduced up to 3 times in 30 minutes MRET-activated water and up to 1.6 times in 60 minutes activated water during the first 6 hours of experiment in aerobic environment (Fig 7 and 8).
Relative Reductant Activity of E.coli bacteria in aerobic environ
This experiment showed that there was no direct correlation between the inhibition of metabolic (reductant) activity and the inhibition of growth of E.coli (bacteriostatic activity) in MRET activated water. The bacteriostatic effect is substantially higher in 60 minutes activated water and the inhibition of reductant activity during the first 3 hours is higher in 30 minutes activated water. Thus, this experiment revealed that the optimum time of activation for the maximum inhibition of metabolic activity of E.coli bacteria in aerobic environment is 30 minutes. The same optimum time of activation was found in the process of another investigation regarding the application of MRET activated water for preventive treatment and enhancement of tumor resistance in vivo on 500 mice for two types of cancer conducted at Kiev Institute of Experimental Pathology, Oncology and Radiobiology of Ukrainian Academy of Science. Taking in consideration that a small population of pathogenic bacteria, such as E.coli, is usually present in complex microbial associations in the intestine of the body, the test on metabolic activity of E.coli bacteria in anaerobic environment was conducted. Anaerobic environment simulates the environmental conditions similar to the conditions in the intestine of humans and animals. The investigation showed that the reductant activity of E.coli bacteria in anaerobic environment practically did not change (Fig 9 and 10).
K Control; 0.5 0.5 hour MRET-activated water; 1.0 1 hour MRET-activated water; KSTER reference to sterility
Reductant Activity of E.coli bacteria in anaerobic environment
This experiment revealed that the process of MRET activation did not have any significant effect on reductant activity of E.coli bacteria in anaerobic environment.
In order to simulate the environmental conditions similar to the conditions in the intestine of humans and animals the test on metabolic activity of microbial associations was conducted in anaerobic environment. It was found that MRET-activated water substantially increased reductant activity of complex microbial associations during the first several hours of experiment (Fig 11).
1.0 1 hour MRET-activated water; 0.5 0.5 hour MRET-activated water; K Control
This experiment revealed that the optimum time of activation for the maximum increase of metabolic activity of microbial associations in anaerobic environment was 30 minutes. The same optimum time of activation was found in the process of inhibition of metabolic activity of E.coli in aerobic environment and in another investigation regarding the application of MRET activated water for preventive treatment and enhancement of tumor resistance in vivo.
This investigation revealed that at low initial concentration of cells of conditionally pathogenic microbiological culture Escherichia coli K-12 in water based nutrient medium activated for 30 minutes and 60 minutes the growth of culture was inhibited 27 and 303 times respectively after the 25 hours of experiment in aerobic environment. This experiment also revealed the strong effect of MRET activated water on the process of division of E.coli microorganisms, the modification of forms of culture cells and the size of colonies. It was observed that one of the reasons of abnormally low growth of E.coli population was related to the modification of the process of cell division in MRET-activated nutrient medium. These results allow admitting that the process of MRET activation and the sterilization effect of MRET water can be applied in food industry and for water purification. The second stage of investigation revealed that the metabolic (reductant) activity of E.coli bacteria reduced up to 3 times in 30 minutes activated water and up to 1.6 times in 60 minutes activated water during the first 6 hours of experiment in aerobic environment. Another experiment showed that the process of MRET-activation did not affect the reductant activity of E.coli bacteria in anaerobic environment and, consequently, should not affect a small population of conditionally pathogenic bacteria, such as E.coli, usually presented in microbial associations in the intestine of the body. In order to simulate the environmental conditions similar to the conditions in the intestine of humans and animals the test on metabolic activity of complex microbial associations was conducted in anaerobic environment. It was discovered that MRET activated process substantially increased reductant activity of complex microbial associations during the first hours of experiment. The same 30 minutes optimum time of activation was observed in the process of inhibition of metabolic activity of conditionally pathogenic E.coli bacteria in aerobic environment and for the maximum increase of metabolic activity of complex microbial associations in anaerobic environment (present in the intestine). The previous investigation regarding the application of MRET activated water for preventive treatment and enhancement of tumor resistance in oncology in vivo on 500 mice also showed the best results on 30 minutes MRET-activated water. Thus, this investigation shows that the ingestion of MRET water is beneficial for the process of digestion and can enhance metabolism of the body. References:
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