Bulletin "Veterinary biotechnology"

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Veterynarna biotekhnolohiia– Veterinary biotechnology, 2023, 43, 9-17 [in Ukrainian]. https://doi.org/10.31073/vet_biotech43-01

BEZYMENNYI M., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it., TARASOV O., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

Institute of Veterinary Medicine of the NAAS

 

THE USE OF A DISINFECTANT IN COMBINATION WITH A GERMICIDE TO NEUTRALIZE BACTERIAL SPORES OF THE GENUS BACILLUS IN THE SOIL

 Introduction. Anthrax is a zoonotic disease caused by the gram-positive bacterium Bacillus anthracis, a pathogen transmitted primarily through soil. The pathogen has been registered on all inhabited continents. The use of germinants is of practical importance to ensure high-quality inactivation of anthrax pathogen spores, since germinated forms are significantly less resistant to environmental conditions and disinfectants compared to spore forms.

Since the problem of anthrax in Ukraine is currently relevant, research on the disinfection of bacterial spores, including the causative agent of anthrax, is important for the successful eradication of pathogens in Ukraine.

The goal of the work. To study the decontamination effectiveness of Bacillus genus bacterial spores using a disinfectant in combination with a germinant.

Materials and methods. The test microorganisms B. cereus ATCC10702, B. anthracis UA 07 (non-pathogenic vaccine strain), Bacillus subtilis 7241 (from the collection of IVM NAAS) were used in the research. A solution of the following composition was used as a germinant: inosine – 0.05%; L-histidine – 0.05%; L-valine – 0.05%; L-serine – 0.03% and water – up to 100%, which specifically affects the germination of Bacillus spores in the soil. The germinant was used in 2 hours exposure, after which the substrate was treated with disinfectants: benzalkonium chloride (alkyl dimethylbenzyl ammonium chloride) (VegaPharm, China); didecyldimethylammonium chloride (VegaPharm, China), and their equal part mixture in a concentration 0,5%.

The required estimated number 5.0×107 CFU/cm3 of microorganisms of each species was separately introduced into the weighing substrate (sterile soil) using a pipette. The control substrate consisted of sterile soil with the addition of Bacillus cereus spores.

The bactericidal effect of the disinfectant was determined by the decrease in the number of microorganisms from the initial values, which was expressed in logarithms (Log). The research was conducted at 25°C, which corresponds to the ambient temperature in warm season.

Statistical processing of the results by descriptive statistics method was performed using Microsoft Excel and Statplus programs.

Results of research and discussion. A significant (р<0.05) reduction of viable bacteria by 5.18±0.4 log (B. cereus ATCC10702) and 4.52±0.3 log (B. anthracis UA 07) was established, respectively, after 12 hours of exposure and 5, 95±0.3 log (B. cereus ATCC10702) and 6.26±0.23 log (B. anthracis UA 07), respectively, after 48 hours of exposure. At the same time, in the control group, the culture growth ranged from 7.25±0.4 to 7.36±0.2 log.

Conclusions and prospects for further research. It has been established that an integrated approach to the disinfection of soil contaminated with spores of Bacillus bacteria, involving the use of a germinant in combination with a disinfectant (quaternary ammonium compounds), provides almost complete neutralization of the pathogen.

Decontamination performed on three soil types against the model microorganism B. cereus ATCC10702 provided a reduction in viable microorganisms from 5.95±0.18 log to 6.15±0.2 log compared to baseline values, and B. anthracis UA 07 – from 5.85±0.3 log to 6.10±0.16 log.

Using this approach will improve and significantly reduce the abundance of pathogenic and opportunistic spore-forming microorganisms in soils.

Keywords: Ваcillus anthracis, spores, germinant, decontamination.

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