Razvoj novih tehnologij za odstranjevanje patogenih mikrobov in toksinov iz različnih vodnih virov (Slovene)

Different pathogenic agents can be present in drinking, irrigation and recreational waters, as a consequence of different factors such as, waste water release, natural disasters, bioterrorism, and/or due to the release of agricultural, industrial and medical residues into water systems. Among these pathogenic agents we can find plant, human and animal viruses and bacteria. One of the key areas within the sustainable development in the increasingly globalized world is the waste water treatment and the management of the treated water effluents or discharges generated in water treatment plants. Such effluents are usually released into environmental waters (fresh or sea water), or alternatively, recycled as irrigation water for agricultural use. Non treated waste water is enriched in toxic heavy metals, as well as pathogenic viruses and bacteria, along with bacterial endotoxins and bacterial DNA. Typical waste water treatment (including sedimentation, active sludge formation, filtration...) does not completely remove the pathogenic viruses and bacteria from the treated water and, therefore, due to health safety reasons, additional disinfection measures need to be applied before using such water for subsequent purposes. Typical disinfection measures are based on chlorine, ozone or UV treatment. Such methods are not efficient against certain pathogens, bacterial endotoxins and bacterial free DNA. In addition, in the case of ozone and chlorine, they result in the formation of hazardous disinfection-by products.   Within the proposed research project we will investigate and evaluate the water disinfection potential of combining two different innovative technologies in order to remove pathogenic bacteria and viruses, as well as bacterial endotoxins and bacterial free DNA. The first technology will be based on the disruption of pathogenic bacteria by irreversible electroporation that consists of Pulsed Electric Field (PEF) application. For effective PEF treatment electric pulse parameters will be optimized in order to achieve the most efficient treatment while minimizing potential toxic product formation that may result from electrode reactions .The second technology will relay on the electrostatic binding of pathogenic viruses and bacterial toxins and DNA to Convective Interaction Media (CIM®) monolithic supports. Monolithic chromatographic supports (developed by the Slovenian company BIA Separations) represent a new generation of chromatographic supports with a structure that allows operating at high flow rates, while at the same time the conferring high dynamic capacity for large molecules. The proposed methods will be optimized with special emphasis on avoiding or minimizing the release or formation of disinfection by-products, and therefore they may constitute a cleaner alternative to classic chemical disinfection methods such as chlorine treatment. The outcome of the project will serve to obtain a proof of principle that will be tested in different water samples, such us waste water treatment plant effluent and drinking water supply, and may be used in future applications of water treatment for removal of pathogenic bio pollutants.   The interdisciplinary project team will include research groups from the Faculty of Electrical Engineering (leaded by Dr Damjan Miklavčič) and the Faculty of Medicine–Institute of Microbiology and Immunology (leaded by Dr Mateja Poljšak-Prijatelj), both from University of Ljubljana, the National Institute of Biology (leaded by Dr Maja Ravnikar) and the Institute of Public Health Maribor (leaded by Dr Maja Rupnik), the high-tech company BIA Separations (leaded by Dr Aleš Štrancar), as a potential end user the waste water treatment plant Domžale (leaded by Dr Marjeta Stražar), and an international project partner Dr Justin Teissie from the Institut de Pharmacologie et de Biologie Structurale (IPBS) CNRS France.

Pulsed electric field and monolith chromatography are methods that are already used in different applications, however in the frame of this project they were used as tools for disruption and/or removal of pathogenic microbes (bacteria and viruses) from water samples. To use these tools in such application, conditions for binding of different viruses on monolithic chromatography columns were optimised with the purpose to enable their efficient binding, concentration, improve their diagnostics and enable their simultaneous removal from water samples. For disruption and / or removal of bacteria from water samples high voltage electrical pulses (in static and flow mode) were used to enable irreversible bacterial lysis and therefore their eradication. The result of the skills and knowledge of all project partners together with the research conducted in the frame of this project resulted in a proof of principle that combines short electrical pulses and CIM monolithic chromatography for purification of waste water effluent which can address issues related to wastewater and drinking water quality as well as recreational waters.

Due to intensive use in agriculture, as well as to other industry factors, the quantity of clean water is rapidly decreasing. People and industries are particularly concerned when affected by natural disasters such as floods, earthquakes or ultimately the uncontrolled use of microorganisms. The technology developed in this project represents a new tool for the treatment of waste water that can be reused in the production of food or irrigation. As such, it is certainly interesting for waste water treatment plants, water supply systems, swimming pools, military missions, etc. The results of the project will certainly be of high interest for a wide professional and laic audience, as well as for the agencies of the ministries of agriculture, environment, health and defence, since the developed technology can be implemented in different segments of the infrastructure for safer use of water resources. Both industrial partners (BIA separations and wastewater treatment plant Domžale) could use the developed techniques and acquired knowledge for the design and testing of a pilot scale device at the wastewater treatment plant Domžale. The proposed innovative approach has the potential for the future production of easy to use equipment which could be marketed globally. In addition, the results of this study are interesting also for all large and small manufacturing companies dealing with waste water treatment and its recirculation in the environment. The knowledge generated during the study was also transferred through the educational process directed to students in the field of biotechnology, electrical engineering, medicine and chemistry. In addition, several doctoral students were working on the project.