The objective of this study was to determine the effect of probiotic preparations contained Bacillus subtilis 109 CFU / ml, Propionibacterium 107 CFU / ml, Lactobacillus plantarum 107 CFU / ml on the physiological state, reproductive functions and the mortality rate of white rats and their offspring. The results of the analysis of the reproductive toxicity of the suspension of strains of probiotics and producers of prebiotics showed that long-term oral administration of the studied doses did not lead to a decrease in the reproductive function of laboratory animals during the experimental period, corresponded to the norm and did not differ significantly from the indicators of the control group. Based on the results obtained, the investigated form of the probiotic can be classified as a drug of low toxicity for reproductive function and, in accordance with GOST 12.1.007-76, referred to the IV hazard class.
We have identified the optimal conditions for the production of chitinolytic enzymes of T.viride in submerged state fermentation. The production of chitinase by a new strain of fungus was carried out on the basal liquid medium, containing (%) colloidal chitin 0.5, NaNO3 0.2, KH2PO4 0.1; MgSO44-7H2O 0.05 and KCl, 0.05. The activity of enzymes of the chitinase complex of the strain was evaluated using the method using dinitrosalicylic acid (DNS reagent). A quantitative determination of the activity of chitinases in a producer microorganism was established by their ability to hydrolyze 0.2% colloidal chitin (in phosphate buffer 0.05 M, pH 5.2), by the content of reducing sugars formed in this process, which were evaluated using a DNS reagent. The results of studies of the influence of various cultivation parameters showed that highest chitinolotic enzymes production by T.viride was obtained at pH 4.0, (301.15-303.15) K and after 144 h growth. The studied soil isolate can be further used in biotechnological research, as well as for biological control of pests and pathogens of agricultural crops.
In this paper, the study of general toxic properties of probiotic preparation based on lactic and propionic acid microorganisms was carried out. Establishment of class of acute toxicity of studied liquid form of the drug was based on fundamental document OECD 423 "Acute Toxic Class Method" (2001). In the study of acute toxicity, the physiological state, growth rate, morbidity and safety, changes in the parenchymal organs of laboratory animals were assessed. The results of determining the acute toxicity of probiotic preparation showed that a single oral administration of it in various doses did not lead to the death of animals during the experimental period, all the main indicators of their vital activity were within the physiological norm and did not differ from the control. The condition of all groups of animals remained satisfactory, no violations of the functional activity of the digestive and urinary systems were revealed. Body temperature, respiration and heart rate of the animals were within normal limits. Shown 100% safety of animals, no toxic signs at 1000-fold increase in the dose of the liquid form of probiotic were not observed. The study of the irritating effect of the drug was carried out on the mucous membrane of the right eye of animals, applying one drop of the sample under the upper eyelid, on the anterior segment of the eye. Consideration of the possible reaction of the mucous membrane of the experimental eye, eyelid and cornea was carried out 15 minutes (fast reaction) after exposure to the drug suspension and after 24, 48 hours of observation (delayed-type hypersensitivity). Evaluation of the effect of the probiotic drug on the conjunctiva showed that the suspension of strains of microorganisms does not cause irritation, both immediately after instillation and throughout the entire study period.
The use of a new form of the probiotic preparation of Bacillus subtilis consisting in emulsion of bacteria in palmitic acid has been studied. When evaluating the effectiveness of probiotics, it was assumed that the content of bacterial DNA in the feces is proportional to their content in the intestine. Analysis of the number of bacterial DNA was carried out by real-time PCR. The results showed a positive effect from the use of emulsified bacteria. It was observed in an increase in the amount of B. subtilis in the intestine and, as a consequence, their DNA in the feces of rats.
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