Project APVV 0016-0171

Project of Slovak Research and Development Agency APVV 0016-0171

Progresive methods for elimination of development and spread of bacterial resistance against clinically relevant antibiotics (duration July 2017-June 2020)

Antibiotic resistance currently represents so serious problem in human and veterinary clinical practice that has become a priority of many important international and world organizations such as FAO, WHO, EFSA, etc. Dissemination and incidence of antibiotic and antibiotic resistant strains in the environment greatly contributes to increased globalization of this problem. Nowadays bacterial resistance associated with a consequent reduction in efficiency and an arsenal of usable antibiotics is not only problem of hospitals. As pointed out by many studies, resistant microorganisms can be found increasingly in wastewater, agriculture, food, as well as in humans or animals. The project therefore aims on analysis of antibiotic resistant bacteria prevalence in selected environments closely related to human activities. Project is targeted also on study of the genetic material related to antibiotic resistance in these environments, as well as on processes supporting an increase in antibiotic resistance. Following the fact that wastewater from hospitals and the community represents a significant reservoir of antibiotic resistant bacteria and resistance genes, the project will focus mainly on the development and monitoring the effectiveness of innovative degradation processes (Fenton reaction, boron-doped diamond electrodes Ferrates) targeted on the elimination of bacteria and genes in the wastewater.

Investigators

Principal investigator:

Investigators:

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Summary of results (2017-2020)

The aim of the project was to analyze antibiotic-resistant bacteria occurrence in various environments covering the entire food chain "from farm to fork" as well as the European strategy One Health. Based on this analysis, the impact of the environment on the antibiotic resistance development was investigated and various technologies were developed to help reduce the overall burden of resistant bacteria. We have detected antibiotic-resistant bacteria throughout the chain, from wastewater, surface water and groundwater, or stabilized sludge used as a fertilizer, through selected foods of plant and animal origin to the intestinal tract of healthy people and domestic or wild animals. An exception was vegetable and fruit salads from fast food and the market in the Slovak Republic. This was also confirmed by our experiments, in which we did not record the transfer of microorganisms from contaminated soil to plants. As the situation was different in the case of vegetable or fruit smoothie drinks, we assume that these foods intended for direct consumption contain resistant bacteria as a result of secondary contamination. We observed the worst situation in the case of effluent samples from medical facilities such as hospitals, in which we detected strains carrying a combination of many resistance genes. If such waters are not treated, they contribute to the spread of resistance not only in municipal wastewater but also at WWTPs and further in the environment. Our research has also shown that various pharmaceuticals or their metabolites with non-antimicrobial activity also contribute to the development of mutations leading to resistance. What is alarming, it is the low concentrations of these substances, in which they commonly occur in wastewater or surface water, that demonstrate this effect on the development of resistance. In most of the studied environments characterized by an increased concentration of heavy metals (landfills from mining and metallurgical production), a statistically significant correlation between increased resistance to metals and resistance to antibiotics was demonstrated. Probably due to the different contents of heavy metals in the studied environments, differences in the observed correlations were observed. The most common link between copper and antibiotic resistance, and in some cases zinc and tetracycline resistance. We tested the effectiveness of advanced technologies to remove resistant bacteria on a variety of wastewater samples, focusing primarily on advanced oxidation processes that they also effectively remove various micropollutants contributing to the development and spread of microbial resistance. The most effective technology was the modified Fenton reaction, which, however, from the point of view of the final treatment of the sample, does not currently appear to be completely suitable for application in practice. Iron-based encapsulated tablets or boron-doped electrodes modified for half energy consumption have proven to be a more suitable alternative. However, due to the unfavorable pandemic situation and the associated limitations, we were not able to bring these experiments and applications into the form of a pilot plant model located at the WWTP. All other set goals of the project were fully fulfilled.

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Results in 2020

BÍROŠOVÁ L., LÉPESOVÁ K., GRABIC R., MACKUĽAK T.: Non-antimicrobial pharmaceuticals can affect the development of antibiotic resistance in hospital wastewater. Environ Sci Pollut Res Int. 2020 Feb 5. doi: 10.1007/s11356-020-07950-x.

MACKUĽAK, T., MEDVECKÁ, E., VOJS STAŇOVÁ, A., BRANDEBUROVÁ, P., GRABIC, R., GOLOVKO, O., MARTON, M., BODÍK, I., MEDVEĎOVÁ, A., GÁL, M., PLANÝ, M., KROMKA, A., ŠPALKOVÁ, V., ŠKULCOVÁ, A., HORÁKOVÁ, I., VOJS, M.: Boron doped diamond electrode – The elimination of psychoactive drugs and resistant bacteria from wastewater. Vacuum 171, 108957, 2020, doi: 10.1016/j.vacuum.2019.108957

BUCKO S., CUVALOVA A., LABUN J., ZBOJOVSKY J., BUJNAKOVA D., KMET V.: Modulation of Staphylococcus aureus biofilm by electromagnetic radiation. J Microbiol Biotechnol Food Sci Sci 9(5), 2020, 1020-1022, doi:10.15414/jmbfs.2020.9.5.1020-1022

GREGOVA G., KMET V. : Antibiotic resistance and virulence of Escherichia coli strains isolated from animal rendering plant. Scientific Reports 10(1), 2020, 17108. doi: 10.1038/s41598-020-72851-5.

TIMKOVA, I., LACHKA, M., KISKOVA, J., MALINICOVA L., NOSALOVA L.,PRISTAS P. , SEDLAKOVA J. : High frequency of antibiotic tolerance in deep subsurface heterotrophic cultivable bacteria from the Rozália Gold Mine, Slovakia. Environ Sci Pollut Res 27, 44036–44044 (2020). https://doi.org/10.1007/s11356-020-10347-5

GERBACOVÁ K., MALINICOVA L., KISKOVA J. ,MASLISOVA V., PRISTAS P. : The Faecal Microbiome of Building-Dwelling Insectivorous Bats (Myotis myotis and Rhinolophus hipposideros) also Contains Antibiotic-Resistant Bacterial Representatives. Curr Microbiol 77, 2333–2344 (2020). https://doi.org/10.1007/s00284-020-02095-z

HAMAROVA L, KOPCAKOVA A, KOCIANOVA-ADAMCOVA M, PIKNOVA M, JAVORSKY P, PRISTAS P. Antimicrobial Resistance of Enterococci from Wild Animals in Slovakia. Polish Journal of Environmental Studies. 30, 2085-2091, (2021), doi:10.15244/pjoes/126371.

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Results in 2019

LISOWSKA-ŁYSIAK K., KOSECKA-STROJEK M., BIAŁECKA J., KASPROWICZ A., GARBACZ K., PIECHOWICZ L., KMET V., SAVINI V., MIĘDZOBRODZKI J.: New Insight into Genotypic and Phenotypic Relatedness of Staphylococcus aureus Strains from Human Infections or Animal Reservoirs. Pol J Microbiol. 68(1),93-104,2019, doi: 10.21307/pjm-2019-011.

HANDROVA L., KMET V.: Antibiotic resistance and virulence factors of Escherichia coli from eagles and goshawks. J Environ Sci Health B. 54(7),605-614, 2019, doi: 10.1080/03601234.2019.1608103

LÉPESOVÁ K., OLEJNÍKOVÁ P., MACKUĽAK T., TICHÝ J., BIROŠOVÁ L.: Annual changes in the occurrence of antibiotic-resistant coliform bacteria and enterococci in municipal wastewater. Environ Sci Pollut Res Int. 26, 18470-18483, 2019, doi: 10.1007/s11356-019-05240-9.

MACKUĽAK T., GRABIC R., ŠPALKOVÁ V., BELIŠOVÁ N., ŠKULCOVÁ A., SLAVÍK O., HORKÝ P., GÁL M., FILIP J., HÍVEŠ J., VOJS M., STAŇOVÁ A.V., MEDVEĎOVÁ A., MARTON M., BIROŠOVÁ L.: Hospital wastewaters treatment: Fenton reaction vs. BDDE vs. ferrate(VI). Environ Sci Pollut Res Int. 26,31812-31821,2019, doi: 10.1007/s11356-019-06290-9

MACKUĽAK T., BRANDEBUROVÁ, A., GRENČÍKOVÁ, A., BODÍK, I., VOJS STAŇOVÁ, A., GOLOVKO, O., KOBA, O., MACKUĽAKOVÁ, M., ŠPALKOVÁ, V., GÁL, M., GRABIC, R.: Music festivals and drugs: Wastewater analysis. Science of the total environmnent 659, 326-334, 2019, doi: 10.1016/j.scitotenv.2018.12.275

ĎURIŠOVÁ, K., BÍROŠOVÁ, L., LÉPESOVÁ, K.: Stationary phase selects predominantly ciprofloxacin resistant strains of Salmonella Typhimurium with mutation in gyrA. J Microbiol Biotechnol Food Sci 9(2), 302-306, 2019, doi: 10.15414/jmbfs.2019.9.2.302-306

LÉPESOVÁ, K., KRAHULCOVÁ, M., MACKUĽAK, T., BÍROŠOVÁ, L.: Sewage sludge as a source of triclosan-resistant bacteria, Acta Chimica Slovaca 12(1), 34-40, 2019, doi: 10.2478/acs-2019-0006

BÍROŠOVÁ, L., SIROTNÁ, Z.: Microbiological risk assessment of sushi food and seafood /Hodnotenie rizika mikrobiálnej kontaminácie potravín typu “sushi” a potravín obsahujúcich plody mora. MPRV SR, e-dokument online- pdf, ISBN 978-80-89738-20-5, 2019, (in Slovak)

MACKUĽAK, T., GÁL, M., BÍROŠOVÁ, L., ČERŇANSKÝ, S., FEHER, M.: Pharmaceuticals, drugs and resistant microorganisms - environmental impact on population health, Current Opinion in Environmental Science & Health 9, 40-48, 2019, doi: 10.1016/j.coesh.2019.04.002

GOŁAWSKA O, ZAJĄC M, MALUTA A, PRISTAS P, HAMAROVÁ Ľ, WASYL D.: Complex bacterial flora of imported pet tortoises deceased during quarantine: Another zoonotic threat? Comp Immunol Microbiol Infect Dis. 65,154-159,2019, doi: 10.1016/j.cimid.2019.05.00

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Results in 2018

LÉPESOVÁ K., KRAKOVÁ L, PANGALLO D., MEDVEĎOVÁ A., OLEJNÍKOVÁ P., MACKUĽAK T., TICHÝ J., GRABIC R., BIROŠOVÁ L. : Prevalence of antibiotic-resistant coliform bacteria, Enterococcus spp. and Staphylococcus spp. in wastewater sewerage biofilm. J Glob Antimicrob Resist. 14:145-151,2018, doi: 10.1016/j.jgar.2018.03.008

KRAHULCOVÁ, M., LÉPESOVÁ, K., BIROŠOVÁ, L.: Occurence of antibiotic resistant bacteria in raw cow milk from vending machines. Acta Chimica Slovaca. 11(1) 55-59, 2018, doi: 10.2478/acs-2018-0009

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Dr. Vladimir Kmet
webpage: http://home.saske.sk/~kmetv/

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