Anti-tuberculosis activity research of 5-(thiophen-2-ylmethyl)- 4H-1,2,4-triazole-3-thiol

Introduction. 1,2,4-Triazole derivatives have already proven themselves to be compounds with low toxicity and high antimicrobial, antifungal, antiviral, hepatoprotective activity. The aim of work is to research the anti-tuberculosis activity of 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazoles-3-thiol on the bacteria strain M. bovis. Materials and methods. At the beginning of the experiment, the effect of the drug concentration and the pH of the medium on the growth rate of the culture at 37 °C was detected. M. bovis 100 passage was selected for this purpose, which was cultured at 37 °C with 5-(thiophen2-ylmethyl)-4H-1,2,4-triazole-3-thiol at the indicated concentrations in a thermostat for 3 months on medium with pH 6.5 (ten test tubes with each medicine concentration) and pH 7.1 (ten test tubes with each medicine concentration). M. bovis 100 passage was used as a control without the addition of 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazole-3-thiol to the medium. Results. The results of the experiment show that the effect of the drug at different concentrations on the medium with a pH 6.5 same with pH 7.1. Growth of pathogenic strain M. bovis 100 passages throughout the observation period (90 days) have been absence for all of the test (0.1 %, 0.5 % and 1.0 %) concentrations of 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazoles-3-thiol. Conclusion. It can be concluded that 0.1 %, 0.5 % and 1.0 % concentration of 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazole-3-thiol influence on the culture properties pathogenic strain M. bovis, which cultured on the medium with pH 6.5 and pH 7.1 at 37 °C, while holding back growth, having a tuberculostatic effect.


Materials and methods
Cultivation and biomass accumulation for strains of M. bovis was carried out on an egg nutrient medium, which was identical in composition to the standard one manufactured by SE "Veterinary Medicine" (Kharkiv, Ukraine). 5-(thiophen-2ylmethyl)-4H-1,2,4-triazole-3-thiol was added to the medium to obtain concentrations 0.1 %, 0.5 %, 1.0 %. Measurement of the required amount of the medicine was performed according to GOST 27025-86. The solutions were prepared according to the methods described in GOST 4212-76 and GOST 4517-87.
Tinctorial properties, morphological features and the timing of primary growth, its intensity and the nature of the subculture of the isolated and accumulated mycobacteria were studied. Number, size, shape, surface, consistency, pigmentation, transparency, gloss and emulsification in saline solution performed the analysis and evaluation of the colonies.
The tinctorial properties of mycobacteria were determined by preparing smears from colonies (cultures) stained with the Zill-Nielsen method and examining them under im-mersion of a SUNNY XS series microscope with a digital microscopic complex "Mykmed-2-1600". The morphology of mycobacteria was determined by length, thickness, shape, nature of cell ends, granularity and location.

Results and discussion
At the beginning of the experiment, the effect of the medicine concentration and the pH of the medium on the growth rate of the culture at 37 °C was detected. M. bovis 100 passage, which was cultured at 37 °C with 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazole-3-thiol at the indicated concentrations in a thermostat for three months on medium with pH 6.5 (ten test tubes with each drug concentration) and pH 7.1 (ten test tubes with each drug concentration) was selected for this purpose. M. bovis 100 passage without the addition of 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazole-3-thiol to the medium have been used as a control.
After these and after cultivation at 37°C, the growth of mycobacterium colonies was recorded and characterized.
As a result, data on the effect of concentrations of 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazole-3-thiol on the cultivation of M. bovis at 37°C on medium with different pH on the culture growth rate were obtained.
Until the 7th day of the experiment, the growth of the culture of M. bovis 100 passage in the control group on medium with a pH 7.1 at 37 °C was not observed.
The results are given in Table 1   The number of single small colonies increased The number of single small colonies increased The number of single small colonies increased Table 3 summarizes the data on the cultural properties of M. bovis 100 passage, which was cultured in a medium with pH 6.5, which contained 5-(thiophen-2-ylmethyl)-4H-1,2,4triazole-3-thiol in three concentrations.
Until the 7th day of the experiment, the growth of the culture of 100 passages of M. bovis on a medium with pH 6.5 at 37 °C was not observed.
The results of the experiment show that the effect of the substance at different concentrations on the medium with pH 6.5 and pH 7.1 are the same. It was marked lack of growth of pathogenic strain M. bovis 100 passages throughout the observation period (90 days) for the all test (0.1 %, 0.5 % and 1.0 %) concentrations of 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazole-3-thiol. Thus, it can be concluded that 0.1 %, 0.5 % and 1.0 % concentration of 5-(thiophen-2-ylmethyl)-4H-1,2,4-triazole-3-thiol actively influence the culture properties pathogenic strain M. bovis cultured on medium with pH 6.5 at 37 0 C, which holding back growth and having a tuberculostatic effect.