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  - 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  - JOUR
AU  - Kolarević, Stoimir
AU  - Micsinai, Adrienn
AU  - Szántó-Egész, Réka
AU  - Lukács, Alena
AU  - Kračun-Kolarević, Margareta
AU  - Lundy, Lian
AU  - Kirschner, Alexander K.T.
AU  - Farnleitner, Andreas H.
AU  - Đukić, Aleksandar
AU  - Čolić, Jasna
AU  - Nenin, Tanja
AU  - Sunjog, Karolina
AU  - Paunović, Momir
PY  - 2021
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4214
AB  - In Serbia less than 13% of collected municipal wastewaters is being treated before their release in the environment. This includes all municipal wastewater discharges from Belgrade (capital city of Serbia; population 1,700,000). Previous research has identified the impacts of raw wastewater discharges from Belgrade on the Danube River, and this study investigated if such discharges also provided a pathway for SARS-CoV-2 RNA material. Samples were collected during the most critical circumstances that occurred so far within the COVID-19 pandemics in Serbia. Grab and composite samples were collected in December 2020, during the peak of the third wave (in terms of reported cases) at the site which receives the wastewater loads in Belgrade. Grab samples collected upstream and downstream of Belgrade were also analyzed. RNA was quantified using RT-qPCR with primer sets targeting nucleocapsid (N1 and N2) and envelope (E) protein genes. SARS-CoV-2 RNA (5.97 × 103 to 1.32 × 104 copies/L) was detected only in samples collected at the site strongly impacted by the wastewaters where all three applied primer sets gave positive signals. Determined concentrations correspond to those reported in wastewater influents sampled at treatment plants in other countries indicating an epidemiological indicator function of used approach for rivers with high pollution loads in countries with poor wastewater treatment.
PB  - Elsevier BV
T2  - Science of The Total Environment
T2  - Science of The Total Environment
T1  - Detection of SARS-CoV-2 RNA in the Danube River in Serbia associated with the discharge of untreated wastewaters
VL  - 783
DO  - 10.1016/j.scitotenv.2021.146967
SP  - 146967
ER  - 

@article{
author = "Kolarević, Stoimir and Micsinai, Adrienn and Szántó-Egész, Réka and Lukács, Alena and Kračun-Kolarević, Margareta and Lundy, Lian and Kirschner, Alexander K.T. and Farnleitner, Andreas H. and Đukić, Aleksandar and Čolić, Jasna and Nenin, Tanja and Sunjog, Karolina and Paunović, Momir",
year = "2021",
abstract = "In Serbia less than 13% of collected municipal wastewaters is being treated before their release in the environment. This includes all municipal wastewater discharges from Belgrade (capital city of Serbia; population 1,700,000). Previous research has identified the impacts of raw wastewater discharges from Belgrade on the Danube River, and this study investigated if such discharges also provided a pathway for SARS-CoV-2 RNA material. Samples were collected during the most critical circumstances that occurred so far within the COVID-19 pandemics in Serbia. Grab and composite samples were collected in December 2020, during the peak of the third wave (in terms of reported cases) at the site which receives the wastewater loads in Belgrade. Grab samples collected upstream and downstream of Belgrade were also analyzed. RNA was quantified using RT-qPCR with primer sets targeting nucleocapsid (N1 and N2) and envelope (E) protein genes. SARS-CoV-2 RNA (5.97 × 103 to 1.32 × 104 copies/L) was detected only in samples collected at the site strongly impacted by the wastewaters where all three applied primer sets gave positive signals. Determined concentrations correspond to those reported in wastewater influents sampled at treatment plants in other countries indicating an epidemiological indicator function of used approach for rivers with high pollution loads in countries with poor wastewater treatment.",
publisher = "Elsevier BV",
journal = "Science of The Total Environment, Science of The Total Environment",
title = "Detection of SARS-CoV-2 RNA in the Danube River in Serbia associated with the discharge of untreated wastewaters",
volume = "783",
doi = "10.1016/j.scitotenv.2021.146967",
pages = "146967"
}

Kolarević, S., Micsinai, A., Szántó-Egész, R., Lukács, A., Kračun-Kolarević, M., Lundy, L., Kirschner, A. K.T., Farnleitner, A. H., Đukić, A., Čolić, J., Nenin, T., Sunjog, K.,& Paunović, M.. (2021). Detection of SARS-CoV-2 RNA in the Danube River in Serbia associated with the discharge of untreated wastewaters. in Science of The Total Environment
Elsevier BV., 783, 146967.
https://doi.org/10.1016/j.scitotenv.2021.146967

Kolarević S, Micsinai A, Szántó-Egész R, Lukács A, Kračun-Kolarević M, Lundy L, Kirschner AK, Farnleitner AH, Đukić A, Čolić J, Nenin T, Sunjog K, Paunović M. Detection of SARS-CoV-2 RNA in the Danube River in Serbia associated with the discharge of untreated wastewaters. in Science of The Total Environment. 2021;783:146967.
doi:10.1016/j.scitotenv.2021.146967 .

Kolarević, Stoimir, Micsinai, Adrienn, Szántó-Egész, Réka, Lukács, Alena, Kračun-Kolarević, Margareta, Lundy, Lian, Kirschner, Alexander K.T., Farnleitner, Andreas H., Đukić, Aleksandar, Čolić, Jasna, Nenin, Tanja, Sunjog, Karolina, Paunović, Momir, "Detection of SARS-CoV-2 RNA in the Danube River in Serbia associated with the discharge of untreated wastewaters" in Science of The Total Environment, 783 (2021):146967,
https://doi.org/10.1016/j.scitotenv.2021.146967 . .