Method of improving growth qualities of the anaerobic nutrient medium with copper nanoparticles
Journal Title: Бюлетень "Ветеринарна біотехнологія" - Year 2017, Vol 31, Issue
Abstract
Introduction. Studying the features of metal nanoparticles influence on metabolic processes of bacterial cells are now an urgent task of medicine, biology and biotechnology since there is little scientific information on the use of metal nanoparticles as stimulators of growth and reproduction of bacterial pathogens.The goal of the work. Determination of the minimum concentration of copper nanoparticles (CuNP) with the highest effect on stimulation of metabolic processes in bacteria C. perfringens (types A, B, C, D), which would ensure the accumulation of the largest volumes of bacterial mass for short cultivation time.Materials and methods. The work was done in the laboratory of anaerobic infections nd. a.Ryzhenka IVM NAAN. Copper nanoparticles synthesized at the F. D. Ovcharenko Institute of Biocolloidal Chemistry of the National Academy of Sciences of Ukraine in the form of a colloidal solution with an initial metal concentration of 32,0 mg / ml and a dimension of 20 nm. The CuNP has a safety passport with tests for the absence of cytotoxicity, genotoxicity and mutagenicity of copper nanoparticles on living objects.The object of the study was suspension of C. perfringens daily cultures (types A, B, C, D) in concentration 2,0 × 109 CFU / cm3.Measurement of the C. perfringens bacterial mass volume was carried out according to the optical standard of turbidity after 6, 12, 24 and 36 hours of cultivation.Used research methods: microbiological, variational-statistical.Results of research and discussion. According to the results of the research, it has been established that activation of the C. perfringens (A, B, C, D types) growth was observed after 6 hours of cultivation in nutrient media regardless of the concentration of CuNP in it. The volume of bacterial mass increased in the 1,7 times (p> 0,001), compared with control indicators.The highest level of growth stimulation and reproduction of C. perfringens bacteria was observed with adding copper nanoparticles to a nutrient medium in concentration 0,008 mg / ml. At such concentration of experimental nanoparticle, the amount of obtained bacterial mass of the pathogen was higher, probably more than 1,9 times (p> 0.001), as compared with control, and was optimal for improvement the growth qualities of the anaerobic nutrient medium.The accumulation of C. perfringens cells after 24 hours of cultivation in the nutrient medium, regardless of concentration CuNP in it, decreased, compared with data obtained after 12 hours of growth, but remained significantly higher at 1,6-1,7 times (p > 0.001) compared with control. After 36 hours of incubation of C. perfringens in nutrient medium, regardless of concentration CuNP in it, the bacterial mass volumes were smaller compared to similar cultivations for 12 hours and were higher in the range of 1,3-1,4 times (p>0,01), compared to control, this indicates acceleration of autolysis of bacterial cells under the influence of their own enzymes.Conclusions and prospects for further researches.The feasibility of using CuNP to improve the growth qualities of the liquid nutrient medium for anaerobes is proved, because copper nanoparticles are components of the oxyreductase enzymes in C. perfringen cells that activate metabolic processes and stimulate their growth and reproduction.The liquid nutrient medium for anaerobes, which included the optimum stimulating dose of CuNP in concentration 0,008 mg / ml, used for cultivation of C. perfringens types A, B, C, D for the period from 12 to 24 hours provided the accumulation of the highest volumes of bacterial mass, which was 1,9 times higher (p> 0,001) than the bacterial growth rates in the control. It indicates a high level of metabolic processes stimulation in the cells of the pathogen by copper nanoparticles of this concentration.Further studies are in development and testing of liquid and solid nutrient medium with adding to it a composite of copper, iron, gold and silver nanoparticles to enhance its growth qualities for the cultivation of anaerobic and aerobic bacteria.
Authors and Affiliations
Yevgen Mintsyuk, Serghiy Nychyk, Olga Gorbatyuk, Galina Ryzhenko, Valentina Andriyaschuk, Alexander Zhovnir, Tatyana Ukhovska, Svetlana Tyutyun
Keywords
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- EP ID EP266498
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