TY  - CONF
AU  - Kolarević, Stoimir
AU  - Micsinai, Adrienn
AU  - Szanto-Egesz, Reka
AU  - Lukacs, Alena
AU  - Kračun-Kolarević, Margareta
AU  - Jovanović Marić, Jovana
AU  - Đorđević, Ana
AU  - Vojnović-Milutinović, Danijela
AU  - Kirschner, Alexander
AU  - Farnleitner, Andreas
AU  - Linke, Rita
AU  - Đukić, Aleksandar
AU  - Kostić, Jovana
AU  - Sunjog, Karolina
AU  - Paunović, Momir
PY  - 2024
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6693
AB  - Pollution of surface waters still represents one of the major environmental challenges in the Republic of Serbia, where wastewaters are discharged directly into the recipients without proper treatment. Within the last decade, we have assessed the microbiological water quality of more than 100 sites situated at different water bodies in Serbia. Quality was determined using faecal indicator bacteria, while microbial source tracking was employed to assess the source of pollution. Almost 50 % of the investigated sites were characterized by critical or even higher level of faecal contamination indicating that untreated wastewaters indeed represent significant pollution pressure on surface waters. Human-associated markers were prevalent in samples from the majority of contaminated sites but the source of pollution was not exclusively human-associated. 
The impact on the water quality of the Danube River was demonstrated in our previous research conducted at the whole river level within the Joint Danube Surveys. In the river stretch from Novi Sad to its confluence with the Velika Morava River, all the midstream samples were critically polluted. In this section, the highest level of pollution was recorded downstream of Belgrade. As ultimate recipients of wastewaters, Danube and its largest tributary Sava currently represent the only solution for disposing of wastewaters originating from the Serbian capital‘s 1,700,000 inhabitants. Such kind of disposal rises additional issues such as antimicrobial resistance and presence of infectious agents in water. For instance during COVID-19 pandemic, we have demonstrated that SARS-CoV-2 RNA can be detected even in surface waters of the Danube River at the sites receiving high wastewater loads from Belgrade which was the unique case for the Basin. 
Despites its extreme importance in this case, wastewater-based epidemiology is neglected in our country and hereby we would like to emphasize the need for implementation of program of such kind in Serbia.
PB  - Belgrade: Serbian Society for Microbiology
C3  - Book of abstracts: 13th Congress of Microbiologists of Serbia: Mikromed Regio 5: From biotechnology to human and planetary health; 2024 Apr 4-6; Belgrade, Serbia
T1  - Impact of untreated wastewaters on the microbiological water quality  of the Danube River  and its tributaries in Serbia
SP  - 65
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6693
ER  - 

@conference{
author = "Kolarević, Stoimir and Micsinai, Adrienn and Szanto-Egesz, Reka and Lukacs, Alena and Kračun-Kolarević, Margareta and Jovanović Marić, Jovana and Đorđević, Ana and Vojnović-Milutinović, Danijela and Kirschner, Alexander and Farnleitner, Andreas and Linke, Rita and Đukić, Aleksandar and Kostić, Jovana and Sunjog, Karolina and Paunović, Momir",
year = "2024",
abstract = "Pollution of surface waters still represents one of the major environmental challenges in the Republic of Serbia, where wastewaters are discharged directly into the recipients without proper treatment. Within the last decade, we have assessed the microbiological water quality of more than 100 sites situated at different water bodies in Serbia. Quality was determined using faecal indicator bacteria, while microbial source tracking was employed to assess the source of pollution. Almost 50 % of the investigated sites were characterized by critical or even higher level of faecal contamination indicating that untreated wastewaters indeed represent significant pollution pressure on surface waters. Human-associated markers were prevalent in samples from the majority of contaminated sites but the source of pollution was not exclusively human-associated. 
The impact on the water quality of the Danube River was demonstrated in our previous research conducted at the whole river level within the Joint Danube Surveys. In the river stretch from Novi Sad to its confluence with the Velika Morava River, all the midstream samples were critically polluted. In this section, the highest level of pollution was recorded downstream of Belgrade. As ultimate recipients of wastewaters, Danube and its largest tributary Sava currently represent the only solution for disposing of wastewaters originating from the Serbian capital‘s 1,700,000 inhabitants. Such kind of disposal rises additional issues such as antimicrobial resistance and presence of infectious agents in water. For instance during COVID-19 pandemic, we have demonstrated that SARS-CoV-2 RNA can be detected even in surface waters of the Danube River at the sites receiving high wastewater loads from Belgrade which was the unique case for the Basin. 
Despites its extreme importance in this case, wastewater-based epidemiology is neglected in our country and hereby we would like to emphasize the need for implementation of program of such kind in Serbia.",
publisher = "Belgrade: Serbian Society for Microbiology",
journal = "Book of abstracts: 13th Congress of Microbiologists of Serbia: Mikromed Regio 5: From biotechnology to human and planetary health; 2024 Apr 4-6; Belgrade, Serbia",
title = "Impact of untreated wastewaters on the microbiological water quality  of the Danube River  and its tributaries in Serbia",
pages = "65",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6693"
}

Kolarević, S., Micsinai, A., Szanto-Egesz, R., Lukacs, A., Kračun-Kolarević, M., Jovanović Marić, J., Đorđević, A., Vojnović-Milutinović, D., Kirschner, A., Farnleitner, A., Linke, R., Đukić, A., Kostić, J., Sunjog, K.,& Paunović, M.. (2024). Impact of untreated wastewaters on the microbiological water quality  of the Danube River  and its tributaries in Serbia. in Book of abstracts: 13th Congress of Microbiologists of Serbia: Mikromed Regio 5: From biotechnology to human and planetary health; 2024 Apr 4-6; Belgrade, Serbia
Belgrade: Serbian Society for Microbiology., 65.
https://hdl.handle.net/21.15107/rcub_ibiss_6693

Kolarević S, Micsinai A, Szanto-Egesz R, Lukacs A, Kračun-Kolarević M, Jovanović Marić J, Đorđević A, Vojnović-Milutinović D, Kirschner A, Farnleitner A, Linke R, Đukić A, Kostić J, Sunjog K, Paunović M. Impact of untreated wastewaters on the microbiological water quality  of the Danube River  and its tributaries in Serbia. in Book of abstracts: 13th Congress of Microbiologists of Serbia: Mikromed Regio 5: From biotechnology to human and planetary health; 2024 Apr 4-6; Belgrade, Serbia. 2024;:65.
https://hdl.handle.net/21.15107/rcub_ibiss_6693 .

Kolarević, Stoimir, Micsinai, Adrienn, Szanto-Egesz, Reka, Lukacs, Alena, Kračun-Kolarević, Margareta, Jovanović Marić, Jovana, Đorđević, Ana, Vojnović-Milutinović, Danijela, Kirschner, Alexander, Farnleitner, Andreas, Linke, Rita, Đukić, Aleksandar, Kostić, Jovana, Sunjog, Karolina, Paunović, Momir, "Impact of untreated wastewaters on the microbiological water quality  of the Danube River  and its tributaries in Serbia" in Book of abstracts: 13th Congress of Microbiologists of Serbia: Mikromed Regio 5: From biotechnology to human and planetary health; 2024 Apr 4-6; Belgrade, Serbia (2024):65,
https://hdl.handle.net/21.15107/rcub_ibiss_6693 .

TY  - JOUR
AU  - Schachner-Gröhs, Iris
AU  - Koller, Michael
AU  - Leopold, Melanie
AU  - Kolm, Claudia
AU  - Linke, Rita
AU  - Jakwerth, Stefan
AU  - Kolarević, Stoimir
AU  - Kračun-Kolarević, Margareta
AU  - Kandler, Wolfgang
AU  - Sulyok, Michael
AU  - Vierheilig, Julia
AU  - Toumi, Marwene
AU  - Farkas, Rózsa
AU  - Toth, Erika
AU  - Kittinger, Clemens
AU  - Zarfel, Gernot
AU  - Farnleitner, Andreas
AU  - Kirschner, Alexander
PY  - 2024
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6581
AB  - The global spread of antimicrobial resistance (AMR) in the environment is a growing health threat. Large rivers are of particular concern as they are highly impacted by wastewater discharge while being vital lifelines serving various human needs. A comprehensive understanding of occurrence, spread and key drivers of AMR along whole river courses is largely lacking. We provide a holistic approach by studying spatiotemporal patterns and hotspots of antibiotic resistance genes (ARGs) along 2311 km of the navigable Danube River, combining a longitudinal and temporal monitoring campaign. The integration of advanced faecal pollution diagnostics and environmental and chemical key parameters allowed linking ARG concentrations to the major pollution sources and explaining the observed patterns. Nine AMR markers, including genes conferring resistance to five different antibiotic classes of clinical and environmental relevance, and one integrase gene were determined by probe-based qPCR. All AMR targets could be quantified in Danube River water, with intI1 and sul1 being ubiquitously abundant, qnrS, tetM, blaTEM with intermediate abundance and blaOXA-48like, blaCTX−M-1 group, blaCTX−M-9 group and blaKPC genes with rare occurrence. Human faecal pollution from municipal wastewater discharges was the dominant factor shaping ARG patterns along the Danube River. Other significant correlations of specific ARGs were observed with discharge, certain metals and pesticides. In contrast, intI1 was not associated with wastewater but was already established in the water microbiome. Animal contamination was detected only sporadically and was correlated with ARGs only in the temporal sampling set. During temporal monitoring, an extraordinary hotspot was identified emphasizing the variability within natural waters. This study provides the first comprehensive baseline concentrations of ARGs in the Danube River and lays the foundation for monitoring future trends and evaluating potential reduction measures. The applided holistic approach proved to be a valuable methodological contribution towards a better understanding of the environmental occurrence of AMR.
PB  - Elsevier
T2  - Water Research
T1  - Linking antibiotic resistance gene patterns with advanced faecal pollution assessment and environmental key parameters along 2300 km of the Danube River
VL  - 252
DO  - 10.1016/j.watres.2024.121244
SP  - 121244
ER  - 

@article{
author = "Schachner-Gröhs, Iris and Koller, Michael and Leopold, Melanie and Kolm, Claudia and Linke, Rita and Jakwerth, Stefan and Kolarević, Stoimir and Kračun-Kolarević, Margareta and Kandler, Wolfgang and Sulyok, Michael and Vierheilig, Julia and Toumi, Marwene and Farkas, Rózsa and Toth, Erika and Kittinger, Clemens and Zarfel, Gernot and Farnleitner, Andreas and Kirschner, Alexander",
year = "2024",
abstract = "The global spread of antimicrobial resistance (AMR) in the environment is a growing health threat. Large rivers are of particular concern as they are highly impacted by wastewater discharge while being vital lifelines serving various human needs. A comprehensive understanding of occurrence, spread and key drivers of AMR along whole river courses is largely lacking. We provide a holistic approach by studying spatiotemporal patterns and hotspots of antibiotic resistance genes (ARGs) along 2311 km of the navigable Danube River, combining a longitudinal and temporal monitoring campaign. The integration of advanced faecal pollution diagnostics and environmental and chemical key parameters allowed linking ARG concentrations to the major pollution sources and explaining the observed patterns. Nine AMR markers, including genes conferring resistance to five different antibiotic classes of clinical and environmental relevance, and one integrase gene were determined by probe-based qPCR. All AMR targets could be quantified in Danube River water, with intI1 and sul1 being ubiquitously abundant, qnrS, tetM, blaTEM with intermediate abundance and blaOXA-48like, blaCTX−M-1 group, blaCTX−M-9 group and blaKPC genes with rare occurrence. Human faecal pollution from municipal wastewater discharges was the dominant factor shaping ARG patterns along the Danube River. Other significant correlations of specific ARGs were observed with discharge, certain metals and pesticides. In contrast, intI1 was not associated with wastewater but was already established in the water microbiome. Animal contamination was detected only sporadically and was correlated with ARGs only in the temporal sampling set. During temporal monitoring, an extraordinary hotspot was identified emphasizing the variability within natural waters. This study provides the first comprehensive baseline concentrations of ARGs in the Danube River and lays the foundation for monitoring future trends and evaluating potential reduction measures. The applided holistic approach proved to be a valuable methodological contribution towards a better understanding of the environmental occurrence of AMR.",
publisher = "Elsevier",
journal = "Water Research",
title = "Linking antibiotic resistance gene patterns with advanced faecal pollution assessment and environmental key parameters along 2300 km of the Danube River",
volume = "252",
doi = "10.1016/j.watres.2024.121244",
pages = "121244"
}

Schachner-Gröhs, I., Koller, M., Leopold, M., Kolm, C., Linke, R., Jakwerth, S., Kolarević, S., Kračun-Kolarević, M., Kandler, W., Sulyok, M., Vierheilig, J., Toumi, M., Farkas, R., Toth, E., Kittinger, C., Zarfel, G., Farnleitner, A.,& Kirschner, A.. (2024). Linking antibiotic resistance gene patterns with advanced faecal pollution assessment and environmental key parameters along 2300 km of the Danube River. in Water Research
Elsevier., 252, 121244.
https://doi.org/10.1016/j.watres.2024.121244

Schachner-Gröhs I, Koller M, Leopold M, Kolm C, Linke R, Jakwerth S, Kolarević S, Kračun-Kolarević M, Kandler W, Sulyok M, Vierheilig J, Toumi M, Farkas R, Toth E, Kittinger C, Zarfel G, Farnleitner A, Kirschner A. Linking antibiotic resistance gene patterns with advanced faecal pollution assessment and environmental key parameters along 2300 km of the Danube River. in Water Research. 2024;252:121244.
doi:10.1016/j.watres.2024.121244 .

Schachner-Gröhs, Iris, Koller, Michael, Leopold, Melanie, Kolm, Claudia, Linke, Rita, Jakwerth, Stefan, Kolarević, Stoimir, Kračun-Kolarević, Margareta, Kandler, Wolfgang, Sulyok, Michael, Vierheilig, Julia, Toumi, Marwene, Farkas, Rózsa, Toth, Erika, Kittinger, Clemens, Zarfel, Gernot, Farnleitner, Andreas, Kirschner, Alexander, "Linking antibiotic resistance gene patterns with advanced faecal pollution assessment and environmental key parameters along 2300 km of the Danube River" in Water Research, 252 (2024):121244,
https://doi.org/10.1016/j.watres.2024.121244 . .

TY  - CONF
AU  - Farnleitner, Andreas
AU  - Schachner, Iris
AU  - Jakwerth, Stefan
AU  - Kittinger, Clemens
AU  - Zarfel, Gernot
AU  - Kavka, Gerhard
AU  - Kolarević, Stoimir
AU  - Kračun-Kolarević, Margareta
AU  - Toth, Erika
AU  - Blaschke, Alfred
AU  - Derx, Julia
AU  - Linke, Rita
AU  - Demeter, Katalin
AU  - Savio, Domenico
AU  - Resicher, Georg
AU  - Kir, Alexander
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6596
AB  - Sustainable and target-oriented microbiological water quality management of rivers needs
information on whole river systems, especially if catchments are large and international. The
microbiological water quality of rivers is of uttermost relevance for human health as river water is
used for several purposes (recreation, drinking water production, irrigation). Besides information on
faecal pollution levels, the origin of faecal pollution and the assessment of associated infection- and
health risks, e g. from antimicrobial resistant bacteria (ARB) are of increasing importance. This
contribution summarizes the concepts and main results from the Joint-Danube-Surveys (JDS) and
discusses future challenges and perspectives (pollution-scenario modelling, infection-risk
assessment) for the Danube River concerning its faecal and AMR pollution status.
Between 2001 and 2019, the whole Danube was sampled four times during the JDS 1 to 4. Beside
standard faecal indicator analysis, cutting-edge molecular detection concepts were applied, including
microbial source tracking (MST) markers by quantitative PCR (qPCR) and high-throughput ampliconsequencing of bacterial communities. For AMR-profiling, clinically relevant bacterial species were
isolated and tested for resistances and resistance genes were determined via qPCR.
With this, we could impressively demonstrate that the JDS create the required multi-national “big
picture” of the microbiological pollution status of the Danube River. Harmonised trans-border
microbiological water quality maps for the whole navigable Danube were established. MST marker
analysis elucidated that the main faecal pollution source along the whole river is human wastewater.
Combined analysis of faecal indicators, MST and AMR-profiles provided a solid basis for assessing the
potential health impacts of AMR associated with faecal pollution
PB  - Austrian Committee - International Association for Danube Research
C3  - Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria
T1  - Joint Danube Survey 1 to 4: Concepts, Lessons Learned and Future Visions on Faecal Pollution and Antimicrobial Resistance
SP  - 9
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6596
ER  - 

@conference{
author = "Farnleitner, Andreas and Schachner, Iris and Jakwerth, Stefan and Kittinger, Clemens and Zarfel, Gernot and Kavka, Gerhard and Kolarević, Stoimir and Kračun-Kolarević, Margareta and Toth, Erika and Blaschke, Alfred and Derx, Julia and Linke, Rita and Demeter, Katalin and Savio, Domenico and Resicher, Georg and Kir, Alexander",
year = "2023",
abstract = "Sustainable and target-oriented microbiological water quality management of rivers needs
information on whole river systems, especially if catchments are large and international. The
microbiological water quality of rivers is of uttermost relevance for human health as river water is
used for several purposes (recreation, drinking water production, irrigation). Besides information on
faecal pollution levels, the origin of faecal pollution and the assessment of associated infection- and
health risks, e g. from antimicrobial resistant bacteria (ARB) are of increasing importance. This
contribution summarizes the concepts and main results from the Joint-Danube-Surveys (JDS) and
discusses future challenges and perspectives (pollution-scenario modelling, infection-risk
assessment) for the Danube River concerning its faecal and AMR pollution status.
Between 2001 and 2019, the whole Danube was sampled four times during the JDS 1 to 4. Beside
standard faecal indicator analysis, cutting-edge molecular detection concepts were applied, including
microbial source tracking (MST) markers by quantitative PCR (qPCR) and high-throughput ampliconsequencing of bacterial communities. For AMR-profiling, clinically relevant bacterial species were
isolated and tested for resistances and resistance genes were determined via qPCR.
With this, we could impressively demonstrate that the JDS create the required multi-national “big
picture” of the microbiological pollution status of the Danube River. Harmonised trans-border
microbiological water quality maps for the whole navigable Danube were established. MST marker
analysis elucidated that the main faecal pollution source along the whole river is human wastewater.
Combined analysis of faecal indicators, MST and AMR-profiles provided a solid basis for assessing the
potential health impacts of AMR associated with faecal pollution",
publisher = "Austrian Committee - International Association for Danube Research",
journal = "Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria",
title = "Joint Danube Survey 1 to 4: Concepts, Lessons Learned and Future Visions on Faecal Pollution and Antimicrobial Resistance",
pages = "9",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6596"
}

Farnleitner, A., Schachner, I., Jakwerth, S., Kittinger, C., Zarfel, G., Kavka, G., Kolarević, S., Kračun-Kolarević, M., Toth, E., Blaschke, A., Derx, J., Linke, R., Demeter, K., Savio, D., Resicher, G.,& Kir, A.. (2023). Joint Danube Survey 1 to 4: Concepts, Lessons Learned and Future Visions on Faecal Pollution and Antimicrobial Resistance. in Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria
Austrian Committee - International Association for Danube Research., 9.
https://hdl.handle.net/21.15107/rcub_ibiss_6596

Farnleitner A, Schachner I, Jakwerth S, Kittinger C, Zarfel G, Kavka G, Kolarević S, Kračun-Kolarević M, Toth E, Blaschke A, Derx J, Linke R, Demeter K, Savio D, Resicher G, Kir A. Joint Danube Survey 1 to 4: Concepts, Lessons Learned and Future Visions on Faecal Pollution and Antimicrobial Resistance. in Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria. 2023;:9.
https://hdl.handle.net/21.15107/rcub_ibiss_6596 .

Farnleitner, Andreas, Schachner, Iris, Jakwerth, Stefan, Kittinger, Clemens, Zarfel, Gernot, Kavka, Gerhard, Kolarević, Stoimir, Kračun-Kolarević, Margareta, Toth, Erika, Blaschke, Alfred, Derx, Julia, Linke, Rita, Demeter, Katalin, Savio, Domenico, Resicher, Georg, Kir, Alexander, "Joint Danube Survey 1 to 4: Concepts, Lessons Learned and Future Visions on Faecal Pollution and Antimicrobial Resistance" in Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria (2023):9,
https://hdl.handle.net/21.15107/rcub_ibiss_6596 .

TY  - CONF
AU  - Kolarević, Stoimir
AU  - Adrienn, Micsinai
AU  - Szanto-Egesz, Reka
AU  - Lukacs, Alena
AU  - Kračun-Kolarević, Margareta
AU  - Jovanović Marić, Jovana
AU  - Đorđević, Ana
AU  - Vojnović-Milutinović, Danijela
AU  - Kirschner, Alexander
AU  - Farnleitner, Andreas
AU  - Linke, Rita
AU  - Đukić, Aleksandar
AU  - Kostić, Jovana
AU  - Sunjog, Karolina
AU  - Paunović, Momir
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6591
AB  - Pollution of natural water bodies is one of the biggest challenges in developing countries, such as
Serbia, where wastewater is discharged directly into the recipients without proper treatment and
poses serious threats to surface water quality, general safety and environmental health. Numerous
pathogenic microorganisms, bacteria, viruses, protozoa and helminthes may reach surface waters
and eventually contaminate groundwater. Despites its extreme importance in this case,
implementation of wastewater-based epidemiology is a challenge for developing countries, as the
majority of households are not connected to sewerage systems. Within our research, we have placed
major focus on recipients of wastewaters as an alternative of targeting raw wastewater. During the
fourth COVID-19 wave in Serbia that started in late February 2021 multiple water samples were
collected at 12 sites at Sava and Danube Rivers in the Belgrade city area which are under the impact
of untreated wastewaters.
SARS-CoV-2 RNA was quantified using RT-qPCR with primer sets targeting nucleocapsid (N1 and N2)
and envelope (E) protein genes. Microbiological (standard fecal indicator bacteria and microbial
faecal source tracking markers), epidemiological, physico-chemical and hydro-morphological
parameters were analysed in parallel. Out of 44 samples analyzed, 31 were positive for at least one
of the target regions of SARS-CoV-2. The results indicated that surveillance of SARS-CoV-2 RNA in
surface waters in context with the large amount of epidemiological and environmental metadata can
be used as epidemiological early-warning tool in countries with poor wastewater treatment.
PB  - Austrian Committee - International Association for Danube Research
C3  - Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria
T1  - Applicability of wastewater based epidemiology in countries with poor wastewater treatment – COVID-19 case study in Serbia
SP  - 11
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6591
ER  - 

@conference{
author = "Kolarević, Stoimir and Adrienn, Micsinai and Szanto-Egesz, Reka and Lukacs, Alena and Kračun-Kolarević, Margareta and Jovanović Marić, Jovana and Đorđević, Ana and Vojnović-Milutinović, Danijela and Kirschner, Alexander and Farnleitner, Andreas and Linke, Rita and Đukić, Aleksandar and Kostić, Jovana and Sunjog, Karolina and Paunović, Momir",
year = "2023",
abstract = "Pollution of natural water bodies is one of the biggest challenges in developing countries, such as
Serbia, where wastewater is discharged directly into the recipients without proper treatment and
poses serious threats to surface water quality, general safety and environmental health. Numerous
pathogenic microorganisms, bacteria, viruses, protozoa and helminthes may reach surface waters
and eventually contaminate groundwater. Despites its extreme importance in this case,
implementation of wastewater-based epidemiology is a challenge for developing countries, as the
majority of households are not connected to sewerage systems. Within our research, we have placed
major focus on recipients of wastewaters as an alternative of targeting raw wastewater. During the
fourth COVID-19 wave in Serbia that started in late February 2021 multiple water samples were
collected at 12 sites at Sava and Danube Rivers in the Belgrade city area which are under the impact
of untreated wastewaters.
SARS-CoV-2 RNA was quantified using RT-qPCR with primer sets targeting nucleocapsid (N1 and N2)
and envelope (E) protein genes. Microbiological (standard fecal indicator bacteria and microbial
faecal source tracking markers), epidemiological, physico-chemical and hydro-morphological
parameters were analysed in parallel. Out of 44 samples analyzed, 31 were positive for at least one
of the target regions of SARS-CoV-2. The results indicated that surveillance of SARS-CoV-2 RNA in
surface waters in context with the large amount of epidemiological and environmental metadata can
be used as epidemiological early-warning tool in countries with poor wastewater treatment.",
publisher = "Austrian Committee - International Association for Danube Research",
journal = "Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria",
title = "Applicability of wastewater based epidemiology in countries with poor wastewater treatment – COVID-19 case study in Serbia",
pages = "11",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6591"
}

Kolarević, S., Adrienn, M., Szanto-Egesz, R., Lukacs, A., Kračun-Kolarević, M., Jovanović Marić, J., Đorđević, A., Vojnović-Milutinović, D., Kirschner, A., Farnleitner, A., Linke, R., Đukić, A., Kostić, J., Sunjog, K.,& Paunović, M.. (2023). Applicability of wastewater based epidemiology in countries with poor wastewater treatment – COVID-19 case study in Serbia. in Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria
Austrian Committee - International Association for Danube Research., 11.
https://hdl.handle.net/21.15107/rcub_ibiss_6591

Kolarević S, Adrienn M, Szanto-Egesz R, Lukacs A, Kračun-Kolarević M, Jovanović Marić J, Đorđević A, Vojnović-Milutinović D, Kirschner A, Farnleitner A, Linke R, Đukić A, Kostić J, Sunjog K, Paunović M. Applicability of wastewater based epidemiology in countries with poor wastewater treatment – COVID-19 case study in Serbia. in Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria. 2023;:11.
https://hdl.handle.net/21.15107/rcub_ibiss_6591 .

Kolarević, Stoimir, Adrienn, Micsinai, Szanto-Egesz, Reka, Lukacs, Alena, Kračun-Kolarević, Margareta, Jovanović Marić, Jovana, Đorđević, Ana, Vojnović-Milutinović, Danijela, Kirschner, Alexander, Farnleitner, Andreas, Linke, Rita, Đukić, Aleksandar, Kostić, Jovana, Sunjog, Karolina, Paunović, Momir, "Applicability of wastewater based epidemiology in countries with poor wastewater treatment – COVID-19 case study in Serbia" in Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria (2023):11,
https://hdl.handle.net/21.15107/rcub_ibiss_6591 .

TY  - CONF
AU  - Schachner-Gröhs, Iris
AU  - Kolm, Claudia
AU  - Vierheilig, Julia
AU  - Leopold, Melanie
AU  - Zarfel, Gernot
AU  - Koller, Michael
AU  - Kittinger, Clemens
AU  - Jakwerth, Stefan
AU  - Linke, Rita
AU  - Kolarević, Stoimir
AU  - Kračun-Kolarević, Margareta
AU  - Toth, Erika
AU  - Farnleitner, Andreas
AU  - Kirschner, Alexander
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6590
AB  - Antimicrobial resistance (AMR) represents one of the top ten global public health threats according
to the WHO. The spread of resistances is no longer limited to clinical settings and the natural
environment, especially aquatic ecosystems, are also strongly involved in the global spread. Holistic
as well as quantitative studies are still scarce. We here present for the first time a large scale study
on the Danube River regarding the distribution of AMR along different faecal pollution patterns.
For this study, samples from an international sampling campaign (Joint Danube Survey 4) as well as
additional seasonal samples were screened for the occurrence and origin of faecal pollution and for
the concentrations of nine representative antibiotic resistance genes (ARGs). In addition to water
samples also river biofilms were monitored to account for both, temporal and permanent river
habitats. Analyses were amended by a set of different cultivation-based and non-cultivation-based
microbiological and environmental data to explain the observed pattern in faecal pollution and ARG
concentrations.
Faecal pollution exists and varied strongly along the whole Danube river; hotspots could be identified
downstream of metropolitan areas at which also substantial concentrations of ARGs could be
detected. Quantifying concentrations of ARGs conferring resistance to different antibiotic classes
revealed strong differences between the different genes. Comparing their presence in water and
biofilm samples, allowed to assess how stably present these genes already are in the Danube River
ecosystem.
Environmental surveys are necessary to establish environmental baselines, to be able to guide future
AMR monitoring as well as to enforce useful management strategies. This integrative study therefore
provides first insights in the antimicrobial resistance situation in the most international river in the
world and second largest river in Europe
PB  - Austrian Committee - International Association for Danube Research
C3  - Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria
T1  - Occurrence of antibiotic resistance genes along gradients of faecal pollution in water and biofilm samples from the whole Danube River
SP  - 25
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6590
ER  - 

@conference{
author = "Schachner-Gröhs, Iris and Kolm, Claudia and Vierheilig, Julia and Leopold, Melanie and Zarfel, Gernot and Koller, Michael and Kittinger, Clemens and Jakwerth, Stefan and Linke, Rita and Kolarević, Stoimir and Kračun-Kolarević, Margareta and Toth, Erika and Farnleitner, Andreas and Kirschner, Alexander",
year = "2023",
abstract = "Antimicrobial resistance (AMR) represents one of the top ten global public health threats according
to the WHO. The spread of resistances is no longer limited to clinical settings and the natural
environment, especially aquatic ecosystems, are also strongly involved in the global spread. Holistic
as well as quantitative studies are still scarce. We here present for the first time a large scale study
on the Danube River regarding the distribution of AMR along different faecal pollution patterns.
For this study, samples from an international sampling campaign (Joint Danube Survey 4) as well as
additional seasonal samples were screened for the occurrence and origin of faecal pollution and for
the concentrations of nine representative antibiotic resistance genes (ARGs). In addition to water
samples also river biofilms were monitored to account for both, temporal and permanent river
habitats. Analyses were amended by a set of different cultivation-based and non-cultivation-based
microbiological and environmental data to explain the observed pattern in faecal pollution and ARG
concentrations.
Faecal pollution exists and varied strongly along the whole Danube river; hotspots could be identified
downstream of metropolitan areas at which also substantial concentrations of ARGs could be
detected. Quantifying concentrations of ARGs conferring resistance to different antibiotic classes
revealed strong differences between the different genes. Comparing their presence in water and
biofilm samples, allowed to assess how stably present these genes already are in the Danube River
ecosystem.
Environmental surveys are necessary to establish environmental baselines, to be able to guide future
AMR monitoring as well as to enforce useful management strategies. This integrative study therefore
provides first insights in the antimicrobial resistance situation in the most international river in the
world and second largest river in Europe",
publisher = "Austrian Committee - International Association for Danube Research",
journal = "Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria",
title = "Occurrence of antibiotic resistance genes along gradients of faecal pollution in water and biofilm samples from the whole Danube River",
pages = "25",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6590"
}

Schachner-Gröhs, I., Kolm, C., Vierheilig, J., Leopold, M., Zarfel, G., Koller, M., Kittinger, C., Jakwerth, S., Linke, R., Kolarević, S., Kračun-Kolarević, M., Toth, E., Farnleitner, A.,& Kirschner, A.. (2023). Occurrence of antibiotic resistance genes along gradients of faecal pollution in water and biofilm samples from the whole Danube River. in Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria
Austrian Committee - International Association for Danube Research., 25.
https://hdl.handle.net/21.15107/rcub_ibiss_6590

Schachner-Gröhs I, Kolm C, Vierheilig J, Leopold M, Zarfel G, Koller M, Kittinger C, Jakwerth S, Linke R, Kolarević S, Kračun-Kolarević M, Toth E, Farnleitner A, Kirschner A. Occurrence of antibiotic resistance genes along gradients of faecal pollution in water and biofilm samples from the whole Danube River. in Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria. 2023;:25.
https://hdl.handle.net/21.15107/rcub_ibiss_6590 .

Schachner-Gröhs, Iris, Kolm, Claudia, Vierheilig, Julia, Leopold, Melanie, Zarfel, Gernot, Koller, Michael, Kittinger, Clemens, Jakwerth, Stefan, Linke, Rita, Kolarević, Stoimir, Kračun-Kolarević, Margareta, Toth, Erika, Farnleitner, Andreas, Kirschner, Alexander, "Occurrence of antibiotic resistance genes along gradients of faecal pollution in water and biofilm samples from the whole Danube River" in Conference Book: 44th IAD conference: Tackling Present & Future Environmental Challenges of a European Riverscape; 2023 Feb 6-9; Krems, Austria (2023):25,
https://hdl.handle.net/21.15107/rcub_ibiss_6590 .

TY  - CONF
AU  - Kirschner, Alexander K.T.
AU  - Schachner, Iris
AU  - Jakwerth, Stefan
AU  - Zarfel, Gernot
AU  - Koller, Michael
AU  - Kittinger, Clemens
AU  - Tóth, Erika
AU  - Kolarević, Stoimir
AU  - Kračun-Kolarević, Margareta
AU  - Blaschke, Alfred Paul
AU  - Derx, Julia
AU  - Linke, Rita
AU  - Demeter, Katalin
AU  - Reischer, Georg
AU  - Savio, Domenico
AU  - Farnleitner, Andreas H.
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6232
AB  - Sustainable and target-oriented microbiological water quality management of rivers needs information on
whole river systems, especially if catchments are large and international. The microbiological water quality
of rivers is of uttermost relevance for human health as river water is used for several purposes (recreation,
drinking water production, irrigation). Besides information on faecal pollution levels, the origin of faecal
pollution and the assessment of associated infection- and health risks, e g. from antimicrobial resistant bacteria (ARB) are of increasing importance. This contribution summarizes the concepts and main results from the Joint-Danube-Surveys (JDS) and discusses future challenges and perspectives (pollution-scenario modelling, infection-risk assessment) for the Danube River concerning its faecal and AMR pollution status.
Between 2001 and 2019, the whole Danube was sampled four times during the JDS. Beside standard faecal indicator analysis, cutting-edge molecular detection concepts were applied, including microbial source tracking (MST) markers by quantitative PCR (qPCR) and high-throughput amplicon-sequencing of bacterial communities. For AMR-profiling, clinically relevant bacterial species were isolated and tested for resistances and resistance genes were determined via qPCR.
With this, we could impressively demonstrate that the JDS create the required multi-national “big picture”
of the microbiological pollution status of large rivers. Harmonised trans-border microbiological water quality aps for the whole navigable Danube were established. MST marker analysis elucidated that the main faecal pollution source along the whole river is human wastewater. Combined analysis of faecal indicators, MST and AMR-profiles provided a solid basis for assessing the potential health impacts of AMR associated with faecal pollution.
PB  - Belgrade: Serbian Society of Microbiology
C3  - Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia
T1  - Faecal pollution and antibiotic resistance in the Danube River: Vision, Concepts and Lessons learned from Joint Danube Surveys
SP  - 144
EP  - 145
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6232
ER  - 

@conference{
author = "Kirschner, Alexander K.T. and Schachner, Iris and Jakwerth, Stefan and Zarfel, Gernot and Koller, Michael and Kittinger, Clemens and Tóth, Erika and Kolarević, Stoimir and Kračun-Kolarević, Margareta and Blaschke, Alfred Paul and Derx, Julia and Linke, Rita and Demeter, Katalin and Reischer, Georg and Savio, Domenico and Farnleitner, Andreas H.",
year = "2022",
abstract = "Sustainable and target-oriented microbiological water quality management of rivers needs information on
whole river systems, especially if catchments are large and international. The microbiological water quality
of rivers is of uttermost relevance for human health as river water is used for several purposes (recreation,
drinking water production, irrigation). Besides information on faecal pollution levels, the origin of faecal
pollution and the assessment of associated infection- and health risks, e g. from antimicrobial resistant bacteria (ARB) are of increasing importance. This contribution summarizes the concepts and main results from the Joint-Danube-Surveys (JDS) and discusses future challenges and perspectives (pollution-scenario modelling, infection-risk assessment) for the Danube River concerning its faecal and AMR pollution status.
Between 2001 and 2019, the whole Danube was sampled four times during the JDS. Beside standard faecal indicator analysis, cutting-edge molecular detection concepts were applied, including microbial source tracking (MST) markers by quantitative PCR (qPCR) and high-throughput amplicon-sequencing of bacterial communities. For AMR-profiling, clinically relevant bacterial species were isolated and tested for resistances and resistance genes were determined via qPCR.
With this, we could impressively demonstrate that the JDS create the required multi-national “big picture”
of the microbiological pollution status of large rivers. Harmonised trans-border microbiological water quality aps for the whole navigable Danube were established. MST marker analysis elucidated that the main faecal pollution source along the whole river is human wastewater. Combined analysis of faecal indicators, MST and AMR-profiles provided a solid basis for assessing the potential health impacts of AMR associated with faecal pollution.",
publisher = "Belgrade: Serbian Society of Microbiology",
journal = "Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia",
title = "Faecal pollution and antibiotic resistance in the Danube River: Vision, Concepts and Lessons learned from Joint Danube Surveys",
pages = "144-145",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6232"
}

Kirschner, A. K.T., Schachner, I., Jakwerth, S., Zarfel, G., Koller, M., Kittinger, C., Tóth, E., Kolarević, S., Kračun-Kolarević, M., Blaschke, A. P., Derx, J., Linke, R., Demeter, K., Reischer, G., Savio, D.,& Farnleitner, A. H.. (2022). Faecal pollution and antibiotic resistance in the Danube River: Vision, Concepts and Lessons learned from Joint Danube Surveys. in Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia
Belgrade: Serbian Society of Microbiology., 144-145.
https://hdl.handle.net/21.15107/rcub_ibiss_6232

Kirschner AK, Schachner I, Jakwerth S, Zarfel G, Koller M, Kittinger C, Tóth E, Kolarević S, Kračun-Kolarević M, Blaschke AP, Derx J, Linke R, Demeter K, Reischer G, Savio D, Farnleitner AH. Faecal pollution and antibiotic resistance in the Danube River: Vision, Concepts and Lessons learned from Joint Danube Surveys. in Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia. 2022;:144-145.
https://hdl.handle.net/21.15107/rcub_ibiss_6232 .

Kirschner, Alexander K.T., Schachner, Iris, Jakwerth, Stefan, Zarfel, Gernot, Koller, Michael, Kittinger, Clemens, Tóth, Erika, Kolarević, Stoimir, Kračun-Kolarević, Margareta, Blaschke, Alfred Paul, Derx, Julia, Linke, Rita, Demeter, Katalin, Reischer, Georg, Savio, Domenico, Farnleitner, Andreas H., "Faecal pollution and antibiotic resistance in the Danube River: Vision, Concepts and Lessons learned from Joint Danube Surveys" in Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia (2022):144-145,
https://hdl.handle.net/21.15107/rcub_ibiss_6232 .

TY  - CONF
AU  - Kolarević, Stoimir
AU  - Micsinai, Adrienn
AU  - Szántó-Egész, Réka
AU  - Lukács, Alena
AU  - Kračun-Kolarević, Margareta
AU  - Đorđević, Ana
AU  - Jovanović Marić, Jovana
AU  - Vojnović-Milutinović, Danijela
AU  - Kirschner, Alexander K.T.
AU  - Farnleitner, Andreas H.
AU  - Linke, Rita
AU  - Đukić, Aleksandar
AU  - Kostić, Jovana
AU  - Sunjog, Karolina
AU  - Paunović, Momir
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6235
AB  - BACKGROUND
Prokaryotic assays are of great interest in eco/geno-toxicology as they are short-term, simple and cost effective. Application of strains of Salmonella typhimurium with modifications such as increased cell wall permeability and lack of excision repair system is common for the detection of genotoxicity. Various eukaryotic models have been developed lately, such as fish derived cell lines, which can mimic the response of the aquatic organisms. Considering different level of cell organization among these models, difference in sensitivity is expected, yet only few studies address this topic.
OBJECTIVES
The major objective was to investigate genotoxic potential of extracts of surface water of the Danube
and to compare the results obtained in applied prokaryotic and eukaryotic models.
METHODS
Surface water samples from 24 sites along the Danube River were extracted by LVSPE Horizon sampler providing relative enrichment factor (REF) of the extracts of 25,000x. For representative prokaryotic model the SOS/umuC assay with S. typhimurium TA1535/pSK1002 was used. Eukaryotic model comprised assessment of cytotoxicity by MTS test and assessment of genotoxicity by comet assay in ZFL (ZebraFish Liver) cells.
RESULTS
While none of the tested samples has induced SOS response, the result of MTS assay indicated that 11 of 24 samples were cytotoxic. In comet assay, genotoxic potential was found in 9 of 24 samples. Results indicated higher sensitivity of used eukaryotic model in comparison with prokaryotic. Applied prokaryotic assay is often suggested to be used as prescreening tool in diagnostics of genotoxic potential which by our opinion could be misleading.
PB  - Belgrade: Serbian Society of Microbiology
C3  - Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia
T1  - Wastewater based epidemiology in countries with poor wastewater treatment - SARS-CoV-2 RNA in surface waters
SP  - 614
EP  - 615
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6235
ER  - 

@conference{
author = "Kolarević, Stoimir and Micsinai, Adrienn and Szántó-Egész, Réka and Lukács, Alena and Kračun-Kolarević, Margareta and Đorđević, Ana and Jovanović Marić, Jovana and Vojnović-Milutinović, Danijela and Kirschner, Alexander K.T. and Farnleitner, Andreas H. and Linke, Rita and Đukić, Aleksandar and Kostić, Jovana and Sunjog, Karolina and Paunović, Momir",
year = "2022",
abstract = "BACKGROUND
Prokaryotic assays are of great interest in eco/geno-toxicology as they are short-term, simple and cost effective. Application of strains of Salmonella typhimurium with modifications such as increased cell wall permeability and lack of excision repair system is common for the detection of genotoxicity. Various eukaryotic models have been developed lately, such as fish derived cell lines, which can mimic the response of the aquatic organisms. Considering different level of cell organization among these models, difference in sensitivity is expected, yet only few studies address this topic.
OBJECTIVES
The major objective was to investigate genotoxic potential of extracts of surface water of the Danube
and to compare the results obtained in applied prokaryotic and eukaryotic models.
METHODS
Surface water samples from 24 sites along the Danube River were extracted by LVSPE Horizon sampler providing relative enrichment factor (REF) of the extracts of 25,000x. For representative prokaryotic model the SOS/umuC assay with S. typhimurium TA1535/pSK1002 was used. Eukaryotic model comprised assessment of cytotoxicity by MTS test and assessment of genotoxicity by comet assay in ZFL (ZebraFish Liver) cells.
RESULTS
While none of the tested samples has induced SOS response, the result of MTS assay indicated that 11 of 24 samples were cytotoxic. In comet assay, genotoxic potential was found in 9 of 24 samples. Results indicated higher sensitivity of used eukaryotic model in comparison with prokaryotic. Applied prokaryotic assay is often suggested to be used as prescreening tool in diagnostics of genotoxic potential which by our opinion could be misleading.",
publisher = "Belgrade: Serbian Society of Microbiology",
journal = "Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia",
title = "Wastewater based epidemiology in countries with poor wastewater treatment - SARS-CoV-2 RNA in surface waters",
pages = "614-615",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6235"
}

Kolarević, S., Micsinai, A., Szántó-Egész, R., Lukács, A., Kračun-Kolarević, M., Đorđević, A., Jovanović Marić, J., Vojnović-Milutinović, D., Kirschner, A. K.T., Farnleitner, A. H., Linke, R., Đukić, A., Kostić, J., Sunjog, K.,& Paunović, M.. (2022). Wastewater based epidemiology in countries with poor wastewater treatment - SARS-CoV-2 RNA in surface waters. in Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia
Belgrade: Serbian Society of Microbiology., 614-615.
https://hdl.handle.net/21.15107/rcub_ibiss_6235

Kolarević S, Micsinai A, Szántó-Egész R, Lukács A, Kračun-Kolarević M, Đorđević A, Jovanović Marić J, Vojnović-Milutinović D, Kirschner AK, Farnleitner AH, Linke R, Đukić A, Kostić J, Sunjog K, Paunović M. Wastewater based epidemiology in countries with poor wastewater treatment - SARS-CoV-2 RNA in surface waters. in Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia. 2022;:614-615.
https://hdl.handle.net/21.15107/rcub_ibiss_6235 .

Kolarević, Stoimir, Micsinai, Adrienn, Szántó-Egész, Réka, Lukács, Alena, Kračun-Kolarević, Margareta, Đorđević, Ana, Jovanović Marić, Jovana, Vojnović-Milutinović, Danijela, Kirschner, Alexander K.T., Farnleitner, Andreas H., Linke, Rita, Đukić, Aleksandar, Kostić, Jovana, Sunjog, Karolina, Paunović, Momir, "Wastewater based epidemiology in countries with poor wastewater treatment - SARS-CoV-2 RNA in surface waters" in Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia (2022):614-615,
https://hdl.handle.net/21.15107/rcub_ibiss_6235 .

TY  - CONF
AU  - Schachner, Iris
AU  - Kolm, Claudia
AU  - Vierheilig, Julia
AU  - Savio, Domenico
AU  - Zarfel, Gernot
AU  - Koller, Michael
AU  - Kittinger, Clemens
AU  - Jakwerth, Stefan
AU  - Linke, Rita
AU  - Kolarević, Stoimir
AU  - Kračun-Kolarević, Margareta
AU  - Tóth, Erika
AU  - Farnleitner, Andreas H.
AU  - Kirschner, Alexander K.T.
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6230
AB  - Human-induced antimicrobial resistance is an emerging concern in aquatic environments. Faecal pollution
sources represent potential propagation pathways, however large-scale quantitative studies in whole river
systems are missing. In the course of the Joint Danube Survey, the Danube River was investigated for the
fourth time in summer 2019 from its headwaters to the delta. Microbial-faecal pollution patterns and the
occurrence of selected antibiotic resistance genes (ARGs) were studied. Subsequently, a one-year time series analysis at selected relevant sites allowed to assess temporal variability.
The intestinal bacterium Escherichia coli was quantified according to standard cultivation methods as indicator for total faecal pollution. In general, longitudinal patterns of faecal pollution showed a comparable picture to previous Joint Danube Surveys, with low to moderate pollution in the upper reaches (Germany, Austria) and critical to strong pollution in the middle and lower sections, especially in Serbia. Genetic microbial source tracking methods, determined via quantitative PCR, allowed us to define human faecal pollution as dominant pollution source along the whole Danube. This trend could be generally confirmed during the annual sampling campaign. To estimate the potential of permanent colonialization of faecal microorganisms within the Danube River ecosystem, biofilms as potential reservoirs were studied along the course of the entire river. Here, E.coli were ubiquitously present but highly variable. Ongoing analyses on the occurrence and quantity of selected ARGs will identify hotspots of antimicrobial resistance and their relationship to faecal pollution along the Danube River and thereby contribute to guide future monitoring and management strategies.
PB  - Belgrade: Serbian Society of Microbiology
C3  - Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia
T1  - Faecal pollution as potential driver of antibiotic resistance genes in the Danube River
SP  - 137
EP  - 138
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6230
ER  - 

@conference{
author = "Schachner, Iris and Kolm, Claudia and Vierheilig, Julia and Savio, Domenico and Zarfel, Gernot and Koller, Michael and Kittinger, Clemens and Jakwerth, Stefan and Linke, Rita and Kolarević, Stoimir and Kračun-Kolarević, Margareta and Tóth, Erika and Farnleitner, Andreas H. and Kirschner, Alexander K.T.",
year = "2022",
abstract = "Human-induced antimicrobial resistance is an emerging concern in aquatic environments. Faecal pollution
sources represent potential propagation pathways, however large-scale quantitative studies in whole river
systems are missing. In the course of the Joint Danube Survey, the Danube River was investigated for the
fourth time in summer 2019 from its headwaters to the delta. Microbial-faecal pollution patterns and the
occurrence of selected antibiotic resistance genes (ARGs) were studied. Subsequently, a one-year time series analysis at selected relevant sites allowed to assess temporal variability.
The intestinal bacterium Escherichia coli was quantified according to standard cultivation methods as indicator for total faecal pollution. In general, longitudinal patterns of faecal pollution showed a comparable picture to previous Joint Danube Surveys, with low to moderate pollution in the upper reaches (Germany, Austria) and critical to strong pollution in the middle and lower sections, especially in Serbia. Genetic microbial source tracking methods, determined via quantitative PCR, allowed us to define human faecal pollution as dominant pollution source along the whole Danube. This trend could be generally confirmed during the annual sampling campaign. To estimate the potential of permanent colonialization of faecal microorganisms within the Danube River ecosystem, biofilms as potential reservoirs were studied along the course of the entire river. Here, E.coli were ubiquitously present but highly variable. Ongoing analyses on the occurrence and quantity of selected ARGs will identify hotspots of antimicrobial resistance and their relationship to faecal pollution along the Danube River and thereby contribute to guide future monitoring and management strategies.",
publisher = "Belgrade: Serbian Society of Microbiology",
journal = "Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia",
title = "Faecal pollution as potential driver of antibiotic resistance genes in the Danube River",
pages = "137-138",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6230"
}

Schachner, I., Kolm, C., Vierheilig, J., Savio, D., Zarfel, G., Koller, M., Kittinger, C., Jakwerth, S., Linke, R., Kolarević, S., Kračun-Kolarević, M., Tóth, E., Farnleitner, A. H.,& Kirschner, A. K.T.. (2022). Faecal pollution as potential driver of antibiotic resistance genes in the Danube River. in Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia
Belgrade: Serbian Society of Microbiology., 137-138.
https://hdl.handle.net/21.15107/rcub_ibiss_6230

Schachner I, Kolm C, Vierheilig J, Savio D, Zarfel G, Koller M, Kittinger C, Jakwerth S, Linke R, Kolarević S, Kračun-Kolarević M, Tóth E, Farnleitner AH, Kirschner AK. Faecal pollution as potential driver of antibiotic resistance genes in the Danube River. in Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia. 2022;:137-138.
https://hdl.handle.net/21.15107/rcub_ibiss_6230 .

Schachner, Iris, Kolm, Claudia, Vierheilig, Julia, Savio, Domenico, Zarfel, Gernot, Koller, Michael, Kittinger, Clemens, Jakwerth, Stefan, Linke, Rita, Kolarević, Stoimir, Kračun-Kolarević, Margareta, Tóth, Erika, Farnleitner, Andreas H., Kirschner, Alexander K.T., "Faecal pollution as potential driver of antibiotic resistance genes in the Danube River" in Electronic abstract book: FEMS Conference on Microbiology in association with Serbian Society of Microbiology; 2022 Jun 30 - Jul 2; Belgrade, Serbia (2022):137-138,
https://hdl.handle.net/21.15107/rcub_ibiss_6230 .

TY  - JOUR
AU  - Kolarević, Stoimir
AU  - Micsinai, Adrienn
AU  - Szántó-Egész, Réka
AU  - Lukács, Alena
AU  - Kračun-Kolarević, Margareta
AU  - Đorđević, Ana
AU  - Vojnović-Milutinović, Danijela
AU  - Jovanović Marić, Jovana
AU  - Kirschner, Alexander K T
AU  - Farnleitner, Andreas A H
AU  - Linke, Rita
AU  - Đukić, Aleksandar
AU  - Kostić-Vuković, Jovana
AU  - Paunović, Momir
PY  - 2022
UR  - https://linkinghub.elsevier.com/retrieve/pii/S004896972204061X
UR  - http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=PMC9232394
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5072
AB  - Wastewater-based epidemiology (WBE) surveillance of COVID-19 and other future outbreaks is a challenge for developing countries as most households are not connected to a sewerage system. In December 2019, SARS-CoV-2 RNA was detected in the Danube River at a site severely affected by wastewaters from Belgrade. Rivers are much more complex systems than wastewater systems, and efforts are needed to address all the factors influencing the adoption of WBE as an alternative to targeting raw wastewater. Our objective was to provide a more detailed insight into the potential of SARS-CoV-2 surveillance in Serbian surface waters for epidemiological purposes. Water samples were collected at 12 sites along the Sava and Danube rivers in Belgrade during the fourth COVID-19 wave in Serbia that started in late February 2021. RNA was concentrated using Amicon Ultra-15 centrifugal filters and quantified using RT-qPCR with primer sets targeting nucleocapsid (N1 and N2) and envelope (E) protein genes. Microbiological (faecal indicator bacteria and human and animal genetic faecal source tracking markers), epidemiological, physicochemical and hydromorphological parameters were analysed in parallel. From 44 samples, SARS-CoV-2 RNA was detected in 31, but only at 4 concentrations above the level of quantification (ranging from 8.47 × 103 to 2.07 × 104 gc/L). The results indicated that surveillance of SARS-CoV-2 RNA in surface waters as ultimate recipients could be used as an epidemiological early-warning tool in countries lacking wastewater treatment and proper sewerage infrastructure. The performance of the applied approach, including advanced sampling site characterization to trace and identify sites with significant raw sewage influence from human populations, could be further improved by adaptation of the methodology for processing higher volumes of samples and enrichment factors, which should provide the quantitative instead of qualitative data needed for WBE.
PB  - Amsterdam: Elsevier
T2  - Science of The Total Environment
T1  - Wastewater-based epidemiology in countries with poor wastewater treatment - Epidemiological indicator function of SARS-CoV-2 RNA in surface waters.
VL  - 843
DO  - 10.1016/j.scitotenv.2022.156964
SP  - 156964
ER  - 

@article{
author = "Kolarević, Stoimir and Micsinai, Adrienn and Szántó-Egész, Réka and Lukács, Alena and Kračun-Kolarević, Margareta and Đorđević, Ana and Vojnović-Milutinović, Danijela and Jovanović Marić, Jovana and Kirschner, Alexander K T and Farnleitner, Andreas A H and Linke, Rita and Đukić, Aleksandar and Kostić-Vuković, Jovana and Paunović, Momir",
year = "2022",
abstract = "Wastewater-based epidemiology (WBE) surveillance of COVID-19 and other future outbreaks is a challenge for developing countries as most households are not connected to a sewerage system. In December 2019, SARS-CoV-2 RNA was detected in the Danube River at a site severely affected by wastewaters from Belgrade. Rivers are much more complex systems than wastewater systems, and efforts are needed to address all the factors influencing the adoption of WBE as an alternative to targeting raw wastewater. Our objective was to provide a more detailed insight into the potential of SARS-CoV-2 surveillance in Serbian surface waters for epidemiological purposes. Water samples were collected at 12 sites along the Sava and Danube rivers in Belgrade during the fourth COVID-19 wave in Serbia that started in late February 2021. RNA was concentrated using Amicon Ultra-15 centrifugal filters and quantified using RT-qPCR with primer sets targeting nucleocapsid (N1 and N2) and envelope (E) protein genes. Microbiological (faecal indicator bacteria and human and animal genetic faecal source tracking markers), epidemiological, physicochemical and hydromorphological parameters were analysed in parallel. From 44 samples, SARS-CoV-2 RNA was detected in 31, but only at 4 concentrations above the level of quantification (ranging from 8.47 × 103 to 2.07 × 104 gc/L). The results indicated that surveillance of SARS-CoV-2 RNA in surface waters as ultimate recipients could be used as an epidemiological early-warning tool in countries lacking wastewater treatment and proper sewerage infrastructure. The performance of the applied approach, including advanced sampling site characterization to trace and identify sites with significant raw sewage influence from human populations, could be further improved by adaptation of the methodology for processing higher volumes of samples and enrichment factors, which should provide the quantitative instead of qualitative data needed for WBE.",
publisher = "Amsterdam: Elsevier",
journal = "Science of The Total Environment",
title = "Wastewater-based epidemiology in countries with poor wastewater treatment - Epidemiological indicator function of SARS-CoV-2 RNA in surface waters.",
volume = "843",
doi = "10.1016/j.scitotenv.2022.156964",
pages = "156964"
}

Kolarević, S., Micsinai, A., Szántó-Egész, R., Lukács, A., Kračun-Kolarević, M., Đorđević, A., Vojnović-Milutinović, D., Jovanović Marić, J., Kirschner, A. K. T., Farnleitner, A. A. H., Linke, R., Đukić, A., Kostić-Vuković, J.,& Paunović, M.. (2022). Wastewater-based epidemiology in countries with poor wastewater treatment - Epidemiological indicator function of SARS-CoV-2 RNA in surface waters.. in Science of The Total Environment
Amsterdam: Elsevier., 843, 156964.
https://doi.org/10.1016/j.scitotenv.2022.156964

Kolarević S, Micsinai A, Szántó-Egész R, Lukács A, Kračun-Kolarević M, Đorđević A, Vojnović-Milutinović D, Jovanović Marić J, Kirschner AKT, Farnleitner AAH, Linke R, Đukić A, Kostić-Vuković J, Paunović M. Wastewater-based epidemiology in countries with poor wastewater treatment - Epidemiological indicator function of SARS-CoV-2 RNA in surface waters.. in Science of The Total Environment. 2022;843:156964.
doi:10.1016/j.scitotenv.2022.156964 .

Kolarević, Stoimir, Micsinai, Adrienn, Szántó-Egész, Réka, Lukács, Alena, Kračun-Kolarević, Margareta, Đorđević, Ana, Vojnović-Milutinović, Danijela, Jovanović Marić, Jovana, Kirschner, Alexander K T, Farnleitner, Andreas A H, Linke, Rita, Đukić, Aleksandar, Kostić-Vuković, Jovana, Paunović, Momir, "Wastewater-based epidemiology in countries with poor wastewater treatment - Epidemiological indicator function of SARS-CoV-2 RNA in surface waters." in Science of The Total Environment, 843 (2022):156964,
https://doi.org/10.1016/j.scitotenv.2022.156964 . .

TY  - CHAP
AU  - Kolarević, Stoimir
AU  - Kračun-Kolarević, Margareta
AU  - Jovanović, Jovana
AU  - Ilić, Marija
AU  - Paunović, Momir
AU  - Kostić-Vuković, Jovana
AU  - Martinović, Rajko
AU  - Jokanović, Sandra
AU  - Joksimović, Danijela
AU  - Pešić, Vladimir
AU  - Kirschner, Alexander K. T.
AU  - Linke, Rita
AU  - Ixenmaier, Simone
AU  - Farnleitner, Andreas
AU  - Savio, Domenico
AU  - Reischer, Georg
AU  - Tomić, Nevenka
AU  - Vuković-Gačić, Branka
PY  - 2020
UR  - https://link.springer.com/chapter/10.1007/698_2019_420
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3952
AB  - The literature data on the microbiological water quality of the surface and groundwaters in Montenegro is very scarce. Therefore, this chapter aims to provide an insight in the microbiological water quality of rivers in Montenegro by compiling the data obtained in period 2009–2018 in national monitoring program and the data collected within the Montenegro survey in 2019 with an emphasis on the hotspots of faecal pollution and possible sources of pollution. Despite the high risk that poor implementation of wastewater treatment might represent for Montenegro, the obtained dataset indicates that overall microbiological water quality of rivers in Montenegro is quite good. About 80% of the investigated sites have bathing water quality according to Montenegrin legislation. Only four sites were identified as hotspots of faecal pollution where the human source of pollution prevails.
PB  - Springer
T2  - The Rivers of Montenegro
T2  - The Rivers of Montenegro
T1  - Microbiological Water Quality of Rivers in Montenegro
DO  - 10.1007/698_2019_420
SP  - 135
EP  - 155
ER  - 

@inbook{
editor = "Pešić, Vladimir, Paunović, Momir, Kostianoy, Andrey G.",
author = "Kolarević, Stoimir and Kračun-Kolarević, Margareta and Jovanović, Jovana and Ilić, Marija and Paunović, Momir and Kostić-Vuković, Jovana and Martinović, Rajko and Jokanović, Sandra and Joksimović, Danijela and Pešić, Vladimir and Kirschner, Alexander K. T. and Linke, Rita and Ixenmaier, Simone and Farnleitner, Andreas and Savio, Domenico and Reischer, Georg and Tomić, Nevenka and Vuković-Gačić, Branka",
year = "2020",
abstract = "The literature data on the microbiological water quality of the surface and groundwaters in Montenegro is very scarce. Therefore, this chapter aims to provide an insight in the microbiological water quality of rivers in Montenegro by compiling the data obtained in period 2009–2018 in national monitoring program and the data collected within the Montenegro survey in 2019 with an emphasis on the hotspots of faecal pollution and possible sources of pollution. Despite the high risk that poor implementation of wastewater treatment might represent for Montenegro, the obtained dataset indicates that overall microbiological water quality of rivers in Montenegro is quite good. About 80% of the investigated sites have bathing water quality according to Montenegrin legislation. Only four sites were identified as hotspots of faecal pollution where the human source of pollution prevails.",
publisher = "Springer",
journal = "The Rivers of Montenegro, The Rivers of Montenegro",
booktitle = "Microbiological Water Quality of Rivers in Montenegro",
doi = "10.1007/698_2019_420",
pages = "135-155"
}

Pešić, V., Paunović, M., Kostianoy, A. G., Kolarević, S., Kračun-Kolarević, M., Jovanović, J., Ilić, M., Paunović, M., Kostić-Vuković, J., Martinović, R., Jokanović, S., Joksimović, D., Pešić, V., Kirschner, A. K. T., Linke, R., Ixenmaier, S., Farnleitner, A., Savio, D., Reischer, G., Tomić, N.,& Vuković-Gačić, B.. (2020). Microbiological Water Quality of Rivers in Montenegro. in The Rivers of Montenegro
Springer., 135-155.
https://doi.org/10.1007/698_2019_420

Pešić V, Paunović M, Kostianoy AG, Kolarević S, Kračun-Kolarević M, Jovanović J, Ilić M, Paunović M, Kostić-Vuković J, Martinović R, Jokanović S, Joksimović D, Pešić V, Kirschner AKT, Linke R, Ixenmaier S, Farnleitner A, Savio D, Reischer G, Tomić N, Vuković-Gačić B. Microbiological Water Quality of Rivers in Montenegro. in The Rivers of Montenegro. 2020;:135-155.
doi:10.1007/698_2019_420 .

Pešić, Vladimir, Paunović, Momir, Kostianoy, Andrey G., Kolarević, Stoimir, Kračun-Kolarević, Margareta, Jovanović, Jovana, Ilić, Marija, Paunović, Momir, Kostić-Vuković, Jovana, Martinović, Rajko, Jokanović, Sandra, Joksimović, Danijela, Pešić, Vladimir, Kirschner, Alexander K. T., Linke, Rita, Ixenmaier, Simone, Farnleitner, Andreas, Savio, Domenico, Reischer, Georg, Tomić, Nevenka, Vuković-Gačić, Branka, "Microbiological Water Quality of Rivers in Montenegro" in The Rivers of Montenegro (2020):135-155,
https://doi.org/10.1007/698_2019_420 . .

TY  - JOUR
AU  - Vrzel, Janja
AU  - Vuković-Gačić, Branka
AU  - Kolarević, Stoimir
AU  - Gačić, Zoran
AU  - Kračun-Kolarević, Margareta
AU  - Kostić, Jovana
AU  - Aborgiba, Mustafa
AU  - Farnleitner, Andreas
AU  - Reischer, Georg
AU  - Linke, Rita
AU  - Paunović, Momir
AU  - Ogrinc, Nives
PY  - 2016
UR  - http://linkinghub.elsevier.com/retrieve/pii/S0048969716316667
UR  - https://www.scopus.com/record/display.uri?eid=2-s2.0-84994378206&origin=SingleRecordEmailAlert&txGid=6CE299281CDB840158BFAC52EC5A2E1C.wsnAw8kcdt7IPYLO0V48gA:71#
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2908
AB  - Coupled measurements of nitrate (NO3-), nitrogen (N), and oxygen (O) isotopic composition (δ15NNO3 and δ18ONO3) were used to investigate the sources and processes of N cycling, while the microbial source tracking (MST) method was used to identify microbiological pollution in the surface water of the Sava River Basin (SRB) in autumn in 2014 and 2015 during high and low water discharge. Atmospheric nitrate deposition or nitrate-containing fertilizers were found not to be significant sources of riverine nitrate in the SRB. The ranges of isotope values suggest that NO3- in the SRB derives from soil nitrification, sewage, and/or manure, which were further supported by MST analysis. Microbiological indicators show the existence of hotspots of fecal pollution in the SRB, which are human associated. Long-term observations indicate persistent fecal contamination at selected locations caused by continuous discharge of untreated wastewaters into the SRB.
T2  - Science of The Total Environment
T2  - Science of The Total Environment
T1  - Determination of the sources of nitrate and the microbiological sources of pollution in the Sava River Basin
VL  - 573
DO  - 10.1016/j.scitotenv.2016.07.213
SP  - 1460
EP  - 1471
ER  - 

@article{
author = "Vrzel, Janja and Vuković-Gačić, Branka and Kolarević, Stoimir and Gačić, Zoran and Kračun-Kolarević, Margareta and Kostić, Jovana and Aborgiba, Mustafa and Farnleitner, Andreas and Reischer, Georg and Linke, Rita and Paunović, Momir and Ogrinc, Nives",
year = "2016",
abstract = "Coupled measurements of nitrate (NO3-), nitrogen (N), and oxygen (O) isotopic composition (δ15NNO3 and δ18ONO3) were used to investigate the sources and processes of N cycling, while the microbial source tracking (MST) method was used to identify microbiological pollution in the surface water of the Sava River Basin (SRB) in autumn in 2014 and 2015 during high and low water discharge. Atmospheric nitrate deposition or nitrate-containing fertilizers were found not to be significant sources of riverine nitrate in the SRB. The ranges of isotope values suggest that NO3- in the SRB derives from soil nitrification, sewage, and/or manure, which were further supported by MST analysis. Microbiological indicators show the existence of hotspots of fecal pollution in the SRB, which are human associated. Long-term observations indicate persistent fecal contamination at selected locations caused by continuous discharge of untreated wastewaters into the SRB.",
journal = "Science of The Total Environment, Science of The Total Environment",
title = "Determination of the sources of nitrate and the microbiological sources of pollution in the Sava River Basin",
volume = "573",
doi = "10.1016/j.scitotenv.2016.07.213",
pages = "1460-1471"
}

Vrzel, J., Vuković-Gačić, B., Kolarević, S., Gačić, Z., Kračun-Kolarević, M., Kostić, J., Aborgiba, M., Farnleitner, A., Reischer, G., Linke, R., Paunović, M.,& Ogrinc, N.. (2016). Determination of the sources of nitrate and the microbiological sources of pollution in the Sava River Basin. in Science of The Total Environment, 573, 1460-1471.
https://doi.org/10.1016/j.scitotenv.2016.07.213

Vrzel J, Vuković-Gačić B, Kolarević S, Gačić Z, Kračun-Kolarević M, Kostić J, Aborgiba M, Farnleitner A, Reischer G, Linke R, Paunović M, Ogrinc N. Determination of the sources of nitrate and the microbiological sources of pollution in the Sava River Basin. in Science of The Total Environment. 2016;573:1460-1471.
doi:10.1016/j.scitotenv.2016.07.213 .

Vrzel, Janja, Vuković-Gačić, Branka, Kolarević, Stoimir, Gačić, Zoran, Kračun-Kolarević, Margareta, Kostić, Jovana, Aborgiba, Mustafa, Farnleitner, Andreas, Reischer, Georg, Linke, Rita, Paunović, Momir, Ogrinc, Nives, "Determination of the sources of nitrate and the microbiological sources of pollution in the Sava River Basin" in Science of The Total Environment, 573 (2016):1460-1471,
https://doi.org/10.1016/j.scitotenv.2016.07.213 . .