Development of a complex anti-radiation protective drug

The article reports results of development of a drug for the prevention and treatment of radiation lesions in animals based on radio modified microorganisms E. coli PL-6 and B. bifidum 1. Aimed at target changing of metabolism, bacteria were exposed to gamma irradiation at doses from 1 to 20 kGy, studying the interaction of microorganisms in the consortium, as well as the safety of produced drugs. Irradiation of E. coli at a dose of 2 kGy led to the appearance of individual cells with polymorphism and having a length exceeding the initial size by 3-7 times. The irradiation of B. bifidum 1 at a dose of 4 kGy contributed to the formation for atypical, polychromic cells, multiple darkening of pigment granules and cell fragments as a result of destruction of microorganisms. The original E. coli PL-6 did not produce the enzymes superoxide dismutaseand catalase, but the metabolites of radiomodified strains of E. coli PL-6 (R10) contained 0.97±0.09 m.c.M/g SOD and 27.38±0.59 mcat/g/ml of catalase activity. In the metabolites of radio-resistant bifidobacteria, as compared with the initial ones, a 1.45-fold excess of peroxidase was recorded.


Introduction
Depressive, allergic and immunodeficient states registered in humans and animals in recent decades are largely associated with the changing environmental situation in the world [1]. Environmental factors can cause pathological conditions in which the body cannot cope with the formed oxide radicals [2]. The further existence and development of animals is impossible without the use of therapeutic agents [3]. Chemical compounds were used in past years to protect animals from ecofactors that were toxic with a small dose increase and often had side effects. Recently, more effective [4], and non-toxic [5] medicinal substances of biological nature are being developed.
Microbial preparations in vivo change the functional activity of cells of organs and systems, increasing the body's radioresistance to radiation exposure [6], increasing number of factors and mediators, which stimulate the formation of antibodies, increasing resistance to exogenous infections due to the activation of the phagocytic function of cells of the phagecytic mononuclear system in blood and tissues [7].
Drugs based on substances of microbial origin in an irradiated organism stimulate the regeneration of hematopoietic tissue, spleen, and bone marrow [8]. There are attempts to treat animals with radiation sickness by using vaccines [9].  . bifidum 1 in the process of vital activity produce antibacterial substances, enzymes, antigens, entero-and exotoxins, cytokines, which individually and in combination with each other have radioprotective properties [10].
Most probiotics have a positive effect on the host organism, modifying the metabolic processes occurring in the intestine and, thereby, providing antiallergic and antitoxic effects [11].
The hypothesis for the studies was the reports that in a mixed population of cultures of different species and genera of microorganisms, the mechanism of interaction with each other is different and can lead to an improvement in the biological parameters of the drugs being developed. An example of such an interaction is the drug Bifikol, created by co-cultivation of E. coli PL-6 and B. bifidum 1. This drug has multifunctional properties, increasing the body's resistance to infectious diseases (dysentery, colibacillosis) and non-infectious (radiation sickness) pathology. At the same time, the issue of the influence of ionizing radiation on the radioprotective properties of the components of the Bifikol drug: E. coli PL-6 and B. bifidum 1, both separately and as part of a consortium, remains unresolved [12].
Based on the foregoing, the aim of the research was to develop a complex drug for the prevention and treatment of radiation injuries to animals based on radio modified strains of microorganisms.

Materials and methods
Some series of experiments investigated the structural, functional and biochemical properties of native and exposed to increasing radiation effects of bacteria E. coli PL-6 and B. bifidum 1 studied the possibility of co-cultivation of two different taxonomic species of radio-modified microorganisms, the harmlessness of preparations made from bacteria and obtained consortia based on the original and radiomodified Escherichia and Bifidobacteria.
Blaurock solid and liquid media, meat-and-peptone broth (MPB) and meat-and-peptone agar (MPA) were used for the growth of microorganisms (Federal Center for Toxicological, Radiation and Biological Safety, Kazan, Russia).
To start the experiment, the initial culture strain of E. coli PL-6 was washed out of a test tube with slant agar with saline solution (Federal Center for Toxicological, Radiation and Biological Safety, Kazan, Russia) -0.95% NaCl in H 2 O. Then the culture was applied to the surface of a mattress with MPA and cultured at 37 °C for 1 day. The grown colonies were washed off with saline into a separate container. Using a unipipette, 0.1 cm 3 of cell suspension was transferred into the first well of a 12-well titrator containing 1.0 cm 3 of saline to each well of a row. After mixing the suspension of cells in the first well, 0.1 cm 3 of the suspensions were transferred to the next well. These procedures were performed sequentially up to the last well of the titrator. Using a light microscope with a Goryaev's camera (limited liability partnership 'Minimed', Russia), the number of microorganisms in the last dilutions was counted and, using the calculation method, the content of microbes in a microliter of the matrix solution was determined. The number of microorganisms in the suspension was standardized to 107-108 m.с./ml by sedimentation or dilution.The obtained biomass was poured into five sterile vials (E-1, E-2, E-3, E-4, E-5), 50 ml for each vial.
Monoculture of bifidobacteria (B. bifidum 1) with a content of living microbial cells of at least 107-108 m.c./vial after rehydration under aerobic conditions in 5.0 ml of saline using a sterile syringe were equally distributed into 5 sealed (without oxygen access) vials (B-1, B-2, B-3, B-4, B-5) containing each 50 ml of Blaurocca's liquid medium. Vials with a culture of bifidobacteria were placed in a thermostat at 37 °C for 4 days. After incubation process, the bacterial cells in the flasks were precipitated at 1.5 thousand rpm for 30 minutes, the supernatant was removed, the centrifugate was resuspended first in a small amount of saline and then, by tenfold dilution, selection and counting of cells in the Goryaev's chamber, stabilized to 10 7 -10 8 m.c./ml. Radiation exposure was carried out using a stationary gamma device 'Issledovatel' ('Baltiets', Narva, Estonia) with the power of 60 Co ionizing radiation sources -1.82e -02 A/kg, in dosesfrom 1 to 25 kGy, followed by sowing irradiated crops on nutrient media. E. coli PL-6 and B. bifidum 1 were exposed to radiation up to the inactivation limit at doses from 1 to 4 kGy (B. bifidum 1) and from 1 to 20 kGy (E. coli . Each stage of irradiation, from outcome to irradiation, studied the cultural, morphological and biochemical properties of bacteria: E. coli PL-6 irradiated at a dose of 7.5 kGy (E-1), 12 kGy (E-2), 18 kGy (E-3), 20 kGy (E-4) and B. bifidum 1 irradiated at doses of 1-4 kGy (B-1, B-2, B-3, B-4). For this purpose, we took the samples of cell cultures, stained smears-prints according to Gram, and made bacterial preparations. Radiomodified Escherichia and Bifidobacteria, as well as their non-irradiated analogs, subsequently were used for their joint cultivation. Based on irradiated bacteria E-4 and B-4. A consortium of radiomodified bacteria (K-4) was prepared. Unirradiated bacteria E-5 and B-5 served as the basis for the formation of a consortium of unirradiated microorganisms (K-5). Microorganisms with the last dose of radiation were mixed in a consortium. For this purpose, 1 cm 3 of the contents of vials E-4 and B-4 were placed with sterile syringes into vials with 50 ml of Blaurock's liquid medium with a partial oxygen content (K-4). A similar procedure was carried out with vials E-5 and B-5 (K-5). Vials with a consortium of native E. coli PL-6 and B. bifidum 1 (K-5) and irradiated bacteria E. coli PL-6 and B. bifidum 1 (K-4) in Blaurock's medium were placed in a thermostat at 37 °C on 4 days. After passage in a nutrient medium, the contents of the KB-4 and KB-5 vials were precipitated, the supernatant was removed, and the centrifugate was resuspended in a small amount of the primer medium.
The study of the expressing (enzymatic) activity of microorganisms was carried out by biochemical methods using the spectrophotometric complex 'SP-46' (Open joint stock company 'LOMO', St. Petersburg, Russia).
To determine the peroxidase activity (PA) according to Kushmanova used pyrogallol as an oxidizable substrate, which was reduced to purpurogallin in the oxygen binding reaction. The maximum absorption spectrum of the spectrophotometer was 430 nanometers. The test solution contained 0.8 ml of 0.006 molar sodium phosphate buffer with a pH of 6.8; 0.12 ml of enzymatic (bacterial) extract; 0.5 ml 0.15% H 2 O 2 ; 1.1 ml H 2 O and 0.5 ml 0.003 M pyrogallol. In the control group, the same amount of distilled water was added instead of 0.5 ml of peroxide. The measurement was carried out for 2-3 minutes. The enzyme activity was determined by the formula (1): where A -enzyme activity; D -the optical density; t -time (s); c -concentration. Determination of catalase activity (CAT) according to Korolyuk was based on the ability of hydrogen peroxide to form a stable colored complex with molybdenum salts. To start the oxidation reaction, 0.03% peroxide solution was added to 1 ml of bacterial extract in Tris-HCl buffer (Research Center of Pharmacotherapy, St. Petersburg, Russia). Distilled water was added to the control tube instead of the sample. The reaction was stopped after 10 min by the addition of 4% ammonium molybdate (Biochem, France). The color intensity of the solution was measured with an SP-46 spectrophotometer (Open joint stock company 'LOMO', St. Petersburg, Russia) at a wavelength of 410 nm. Catalase activity was calculated using the formula (2): where: E is the activity of catalase (m.kat/l); A k and A op are the activity of the experimental and control samples; V is the volume of the introduced sample (0.1 ml); t is the incubation time (10 minutes); K is the coefficient of millimolar extraction of hydrogen peroxide, equal to 22 . 10 3 mM -1 cm -1 . The determination of the activity of superoxide dismutase (SOD) according to Paoletti [13] in the modification of our laboratory is based on the detection of the degree of inhibition by superoxide Nucleic acids were determined spectrophotometrically according to Spirin [14], which consists in determining the difference in refraction of suspensions of native and denatured DNA at different wavelengths (limited liability company 'Samson-Med', St. Petersburg, Russia). We used lyophilized sodium salt of calf thymus DNA, NaCl, HClO 4 65%, The selected samples native (K-5) and radiomodified cells (K-4), as well as, consortia were subjected to biological testing for harmlessness.The test substances were injected subcutaneously into the thigh area in mice in a volume of 0.1 cm 3 : The animals of the 13th group were not injected with the drug (biological control group).
The animals were clinically observed for 10 days, noting their viability, behavioral reactions, and the presence of appetite.
After 10 days, the animals were bled from the heart during ethereal euthanasia. The number of formed elements (erythrocytes, leukocytes) was determined using a Goryaev's camera and a microscope (Open joint stock company 'LOMO', St. Petersburg, Russia), hemoglobin -using a Salihemometer (Open joint stock company 'LOMO', St. Petersburg, Russia).
The results and the reliability of the results were assessed by the Student's criterion and described using the Microsoft Excel 2016 software included in the Microsoft Office 2016 software package. The obtained digital material was subjected to statistical processing using the computer program 'Statistika 6'.

Results and discussion
Cell cultures: B. bifidum 1 (after rehydration, passing in the Blaurock medium, precipitation and separation of the supernatant), as well as E. coli PL-6 (after passing mesopatamia on agar and diluting with saline solution to a concentration of 100 million m.c./ml) were used for the production of Gramstained microbial preparations E-5 (Escherichia) and B-5 (bifidobacteria), and were also selected as test microbial preparations for testing their harmlessness on animals. In addition, the original and subsequently radio-modified cultures were studied for their cultural, morphological and biochemical properties. The initial cultures of microorganisms were subjected to increasing irradiation with gamma quanta at doses from 1 to 25 kGy (E. coli PL-6) and from 1 to 4 kGy (B. bifidum 1) with their intermediate passaging on updated nutrient media. At each stage of the work, samples were taken for the above studies (E1-E4 and B1-B4).
During the research, it was found that E. coli PL-6 bacteria have rapid growth on simple and synthetic nutrient media at a temperature of 15 °C to 45 °C at pH of 7.2-7.4, the optimal growth temperature is 37-38 °C. On dense media, microbes formed rounded, convex colonies of medium size, moist, with a smooth, shiny surface with even edges. The growth of microorganisms in liquid media occurred in the form of intense uniform turbidity of the medium, the formation of a precipitate, which disintegrated during shaking, forming a homogeneous suspension. In the Endo environment, microbes formed colonies of red color, mainly with a metallic sheen.
Under microscopy, the bacteria were located singly or in the form of small conglomerates with sizes from 1-3 microns in length and 0.5-1.3 microns in width, with Gram -negative staining. B. bifidum 1 bacteria are strict anaerobes in vivo, but under laboratory conditions they have acquired the ability to develop in the presence of a small amount of oxygen and carbon dioxide. The optimal growth temperature ranged from 37 °C to 41 °C. The optimal pH value is 6-7. Bifidobacteria were grown on liquid and solid nutrient media, creating anaerobic conditions. In the liquid medium of Blaurocca, the culture grew in the form of filamentous colonies located in the lower part of the medium. Shaking the test tube in the nutrient medium led to uniform turbidity. On dense media, bifidobacteria formed colonies of various shapes from flat, hemispherical to shiny, rough with a darker center. The color of the colonies varied from white-gray to dark brown.
Microscopy revealed that bifidobacteria are polymorphic rods with unilateral or bilateral bifurcation, 4-5 microns long, 0.2-0.5 microns thick, arranged in clusters or individual cells. They form bifurcated structures when grown on liquid or nutrient-poor media; rods are formed on solid or nutrient-poor media, located separately or in the form of adhesions with each other.
Irradiated E. coli PL-6 in doses from 1 to 18 kGy per MPB grew in the form of uniform turbidity of the medium. When the bacteria were irradiated at a dose of 20 kGy, the nutrient medium slightly opalesced. Irradiation at a dose of 25 kGy led to complete inactivation of microbes. The nutrient medium remains transparent.
Escherichia, subjected to gamma quantaat a dose of 20 kGy on Mesopotamia agar (MPA), give single colonies. Repeated passaging of cells on renewed media ensures continuous growth of the E. coli (R10) cell colony.
B. bifidum 1 bacteria irradiated at doses of 1 and 2 kGy in the liquid medium of Blaurokk give typical filamentous colonies that visually fill the space of the nutrient medium. Irradiation at a dose of 3kGy suppresses the growth of microorganisms, there is a slowdown in growth, filamentous colonies do not fill the entire space of the medium. Irradiationat a dose of 4 kGy stably suppresses the growth of the culture, while the liquid medium of Blaurokk looks transparent.
Bifidobacteria exposed to radiation at a dose of 4 kGy in a semi-solid Blaurokk medium form single colonies, which, after repeated (R6) passaging in updated media, give a continuous growth of the microbial mass.
When studying microbial preparations, it was found that E. coli PL-6 bacteria irradiated at doses of 2 kGy and higher are characterized by polymorphism. In this case, the individual cells have a length that exceeds the original size by 3-7 times. At a radiation exposure of 12 kGy, the average number of altered (modified) cells in 10 fields of view of the microscope (magnification of 10x100) is ≈1.7. At the same time, in some fields of vision, dense, physiologically inactive, smaller cells in size the dimensions of ≈0.3-0.5 µmwere revealed. After irradiation at a dose of 18 kGy, the number of modified microorganisms increases to 2.8 and 20 kGy=5.1 units.
On microbial preparations made from bacteria irradiated at doses from 10 to 20 kGy, cells with the presence of darkening of the aggregating substance or, conversely, light spaces (vacuoles) caused by the disorganization of the bacterial content, are recorded. The amount of damage increases with increasing radiation dose. On these preparations, the number of viable cells is insignificant.
Studies have shown that under radiation exposure from 2 to 4 kGy on microbial preparations of bifidobacteria, multiple darkening of pigment granules and tissue fragments are recorded as a result of destructive changes in microorganisms. After irradiation of microorganisms at a dose of 4 kGy, atypical, increased in length, polychromic cells are revealed.
At the second stage of the experiment, the radiomodified bacteria E. coli (R10) and B. bifidum (R6) (E-4 and B-4) were co-cultivated in a liquid Blaurock medium with a partial presence of oxygen (K-4). A consortium of unirradiated bacteria E. coli PL-6 and B. Bifidum-1 (K-5) was studied as a control to compare the morphological properties of these cultures. The incubation lasted 4 days. At the same time, microbial preparations were prepared every day for the morphological assessment of microorganisms, and after 4 days -microbial preparations (K-4 and K-5) to determine the harmlessness of the microbial mass for animals. A day after the start of incubation of consortia of irradiated bacteria and their unirradiated analogs (K-4 and K-5), a significant number of active Escherichiosis rods surrounded by single bifidobacteria and granules of destroyed B. Bifidum cells were found on microbial preparations.
After 2 days, the number of Escherichia was significant; some of the E. coli cells were dense, physiologically inactive. Physiologically active bifidobacteria were present on the preparations.
After 3 days, the number of active Escherichia was insignificant, destroyed cells were revealed on the preparations, part of the Escherichia was in the stage of statis. Many active bifidobacteriahave been identified.
After 4 days, the number of bifidobacteria prevailed over the cells of Escherichia coli. The consortia of radiomodified and non-irradiated bacteria did not differ in the characteristics of interaction, but the cellular reactions of the irradiated cells proceeded more intensively, which was characterized by an accelerated transition from aerobic culture (E. coli) to anaerobic culture (B.
In radio-resistant bifidobacteria, as compared to native bacteria, a 1.45 times higher peroxidase content was recorded in the culture liquid.
The prepared bacterial suspensions of native Escherichia and bifidobacteria (E-5, B-5) and radiomodified variants of E. coli (E-1, E-2, E-3, E-4) and B. bifidum 1 (B-1, B -2, B-3, B-4), as well as their native (K-5) and radiomodified (K-4) consortia in the next series of experiments were subjected to biological testing for harmlessness. As a biological model for testing, 39 white mice with an average live weight of 20±2 g were used, divided according to the principle of analogs into 13 groups, three heads in each group.
It was found that the survival rate of mice during the use of microbial suspensions was 100%, and the animals of all groups during the entire period looked clinically healthy, were mobile, and retained their appetite. There were no differences in the behavioral responses of individuals who received different types of cells with different physical effects. All animals remained viable 10 days after the initiation of safety testing of the study drugs.The results of hematological studies are shown in figure 1 and table 1. From these tables and the figure it follows that the studied microbial preparations did not have a significant effect on the hematological parameters of animals. There are also insignificant differences with biological control (BC) in animals of groups E-2, E-3 and K-4 an increase in the content of erythrocytes by 12-13% on the 10th day. In mice from groups E-1, E-2, E-3 and K-4 -leukocytes level by 11-13%, in mice from groups E-2, E-3 and K-4 -hemoglobin by 11-15 %.
Thus, sequential irradiation of E. coli PL-6 and B. bifidum 1 cultures in increasing doses led to an increase in radioresistance with induction of enhanced synthesis of radioprotective enzymes superoxide dismutase, catalase, and peroxidase activity involved in the formation of the organism resistance to lethal effects of ionizing radiation.
At the next stage of the experiments, the obtained microbial substances were used to design radioprotective drugs.
The main purpose of the experiments was a directed change in the phenotype of bacteria of E. coli PL-6 and B. bifidum 1 strains towards an increase in the expression of some factors and mediators useful from the point of view of practice -stimulating metabolism, radioprotective, antioxidant, antitoxic and others, which include enzymes, antigens, entero-, exotoxins, cytokines [15]. The main tool for such an effect was ionizing radiation using a stationary gamma apparatus 'Issledovatel' with 60 Co sources ('Baltiets' (Narva). Irradiation was carried out in the dose range from 1 to 25 kGy with a step of 1 kGy, then the irradiated culture was sown on nutrient media and was assessed by the growth of colony-forming units.
Used strain E. coli PL-6. Under irradiation at a dose of 20 kGy, changes in the cultural, morphological and biochemical properties of bacteria were noted. After irradiation at a dose of 25 kGy, all cells died. In this regard, bacteria with an irradiation dose of 20 kGy were used to form a microbial consortium. A similar situation was observed after irradiation of B. bifidum 1 bacteria, but the doses of gamma irradiation were significantly lower.
Irradiation of the initial E. coli PL-6 strain with successively increasing doses yielded a radioresistant E. coli PL-6 (R10) strain, which differed from the initial survival rate at a dose 2.14 times higher than the initial level, and a stable radioresistant variant of bifidobacteria B bifidum 1 (R6) strain, which differs from the initial one in high radioresistance and survives at a dose 2.5 times higher than the initial level.
According to S Bourdouxet al. [14] resistance to drying and radioresistance are similar, as they cause similar cellular damage. The authors found that microorganisms isolated from the Dry Valleys of Antarctica gave growth of cultured cells after exposure to ionizing radiation of 4 and 6 kGy. It was shown that cooling bacteria to -79 °C increased the radiation resistance of microorganisms by 9 times [16].
It was found [17] that microorganisms living at extremely high temperatures of geysers and thermal springs have different mechanisms of adaptation to stress, which allow them to overcome numerous physical and chemical barriers to survival, such as DNA damage, oxidative explosions and protein damage. These bacteria have been used to isolate the radiation-resistant thermophiles Deinococcus geothermalis, which are also resistant to desiccation and maintain their homeostasis through advanced DNA repair mechanisms, the reactive oxygen species (ROS) detoxification system, and the accumulation of compatible solutes.
Studied [18] cyanobacteria dominating in the most extreme arid places of hot and cold deserts. After irradiation of 10 different strains of Chroococcidiopsis at a dose of 2.5 kGy, the survival rate was 35-80%, with irradiation of the 4 most radioresistant bacteria at a dose of 5 kGy, the survival rate decreased by 1-2 orders of magnitude, but viable cells were restored after irradiation with 15 kGy, the dose 20 kGy was fatal [19] showed that radiation tolerance in Chroococcidiopsis is associated with a decrease in oxygen exposure upon drying.
A number of researchers have shown that the evolution of microorganisms includes the simultaneous spread of various beneficial mutations [20].
Experiments with the effect of low-energy electrons and gamma radiation on DNA molecules of prokaryotes are described [21]. It has been shown that radiation reduces the viability of cells, and DNA molecules are a target for radiation. DNA repair defects cause increased sensitivity to DNA damaging agents, accumulation of mutations in the genome, and, ultimately, the development of metabolic disorders [22]. However, cells have at least seven mechanisms for restoring the structural integrity of DNA, and in the case of irreparable damage, they trigger the mechanism of apoptosis [23].
The radioactive effect on bacteria Staphylococcus epidermidis and E. coli has been studied [24], and it has been shown that the growth of microorganisms in an aerobic environment generates reactive oxygen species (ROS), which leads to oxidative stress. Antioxidant enzymes (superoxide dismutases and hydroperoxidases) and DNA repair mechanisms provide protection against ROS. Acid stress is associated with the induction of Mn superoxide dismutase (MnSOD) in Lactococcuslactis and Staphylococcus aureus. In addition, these results were confirmed in Escherichia coli strains lacking both MnSOD and iron SOD (FeSOD), but expressing heterologous MnSOD from S. thermophilus. It has been found that in E. coli FeSOD does not provide the same protection as MnSOD, and hydroperoxidases are equally important in protecting cells from acid stress. These data explain the ability of some microorganisms to survive better in an acidified environment [25]. The key antioxidant in the metabolism of long-chain fatty acids (LCFA) in E. coli is the coenzyme benzoquinone [26]. With the help of external gamma radiation [27], as a result of directed evolution, an extremely stable strain of E. coli was created. Were obtained four populations of E. coli, each of which is specially adapted to survive under the influence of high doses of ionizing radiation. Several mutations in the RecA gene and deletion of the e14 prophage contributed to the creation of a new phenotype [28].
Based on the foregoing, the use of physical methods is a tool for changing the phenotypic properties of bacteria and the developmentof anti-radiation drugs for animals. Our data are consistent with the studies of other authors and can serve as a basis for further experiments.

Conclusion
The radiation stress caused by the bacteria E. coli PL-6 and B. bifidum 1 morphological changes in the appearance of atypical cells with polymorphism, with a length larger than the original size from 3 to 7 times the number is less than 1% of B. bifidum 1 after a dose of radiation exposure of 4 kGy; 1.7% with irradiation of E. coli PL-6 in a dose of 12 kGy and 2.8% after radiation effects on cells at a dose of 18 kGy and 5.1% after irradiation of E. coli PL-6 in a dose of 20 kGy.
Irradiation of E. coli bacteria at a dose of 25 kGy and B. bifidum 1 at a dose of 5 kGy led to complete sterilization of microorganisms. The joint cultivation of E. coli PL-6 and B. bifidum 1 bacteria in the consortium was expressed by the manifestation of cell antagonism, in which certain microorganisms prevailed in the most favorable conditions for each type of cell, namely: in the initial stage in the presence of oxygen (1-2 days) -E. coli PL-6 bacteria and later, in an airless environment (3-4 days) -B.bifidum.
Biochemical studies revealed a change in the morphological and expressing properties of radiomodified bacteria compared to their non-irradiated analogues in the form of an increase in the content of the amount of deoxyribonucleic acid in E. coli bacteria-by 2.51 times, an increase in peroxidase activity -by 1.67 times, the appearance in escherichia of the properties of the production of superoxide dismutase enzymes -0.97±0.09 m.c.M/g, catalase -27.38±0.59 mcat/g/ml; in B. bifidum, the increase in peroxidase expression was 1.45 times.
Preparations made from radiomodified cells did not have a negative effect on the tested laboratory animals, causing only minor changes in the picture of their blood -the content of the absolute number of red blood cells, white blood cells and hemoglobin.