Determination of the level and source of microbial pollution in the Sava River Basin

Introduction. The contamination of water bodies by sewage or manure is generally determined by fecal indicator bacteria. Specific differentiation between sources of contamination is of particular importance, since the health risk to humans is usually considered higher from human, than from animal fecal contamination. Quantitative PCR (qPCR)-based assays for analysis of human- or animal-associated genetic Bacteroidetes fecal markers have gained increased popularity in the field of microbial source tracking (MST). Aim. The aim of the present study was to assess the level and source of microbial pollution in the Sava River Basin (SRB). Materials and methods. The sampling at the SRB was performed in September 2015, at 15 locations. Additional samples were collected from 4 wastewater outlets. Quantification of total coliforms, E. coli and enterococci was performed with Colilert Quanti-Tray 2000 and MPN approach. The human-associated BacHum and HF183II, the ruminant-associated BacR and the pig-associated Pig2Bac fecal markers were enumerated by quantitative PCR (qPCR). Results. According to the concentrations of fecal indicator bacteria, the river samples were scattered within the classes I and II (slight to moderate pollution). According to all monitored indicators, samples collected at 3 wastewater outlets were excessively contaminated. The results of MST revealed the presence of human-associated fecal markers BacHum and HF183II at 13 locations. The ruminant-associated BacR and the pig-associated Pig-2-Bac markers were not detected. High correlation was observed between the standard fecal indicators and human associated fecal markers. Conclusions. The results reveal human origin of fecal pollution in the SRB and indicates the urgent need for effective wastewater treatment plants in water management

Evaluation of genotoxic pressure along the sava river

©2016 Kolarević et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. In this study we have performed a comprehensive genotoxicological survey along the 900 rkm of the Sava River. In total, 12 sites were chosen in compliance with the goals of GLOBAQUA project dealing with the effects of multiple stressors on biodiversity and functioning of aquatic ecosystems. The genotoxic potential was assessed using a complex battery of bioassays performed in prokaryotes and aquatic eukaryotes (freshwater fish). Battery comprised evaluation of mutagenicity by SOS/umuC test in Salmonella typhimuriumTA1535/ pSK1002. The level of DNA damage as a biomarker of exposure (comet assay) and biomarker of effect (micronucleus assay) and the level of oxidative stress as well (Fpg-modified comet assay) was studied in blood cells of bleak and spirlin (Alburnus alburnus/ Alburnoides bipunctatus respectively). Result indicated differential sensitivity of applied bioassays in detection of genotoxic pressure. The standard and Fpg-modified comet assay showed higher potential in differentiation of the sites based on genotoxic potential in comparison with micronucleus assay and SOS/umuC test. Our data represent snapshot of the current status of the river which indicates the presence of genotoxic potential along the river which can be traced to the deterioration of quality of the Sava River by communal and industrialwastewaters. The major highlight of the study is that we have provided complex set of data obtained from a single source (homogeneity of analyses for all samples)

Impact of pollution on rivers in Montenegro: Ecotoxicological perspective

Montenegrin surface water and groundwater are important for the Balkan Peninsula since they are connected by the transboundary Dinaric Karst Aquifer System with the waters of additional five countries. The pollution from the surface water can rapidly infiltrate in aquifer and endanger this sensible ecosystem and the health of humans through drinking water supply. This chapter gives insights in the pressures of pollution on Montenegrin waters and in a limited literature data regarding freshwater ecotoxicological studies in Montenegro. Also, this chapter provides new ecotoxicological data obtained during survey in 2019, with a focus on the sites which are identified as hotspots of fecal pollution. The highest responses of biomarkers which indicate embryotoxic, genotoxic, and phytotoxic effects in zebrafish embryo test and in roots of Allium cepa were obtained at Ćehotina – downstream of Pljevlja. Similar results were detected at the site downstream Mojkovac at Tara, yet this site is affected by different type of pollution. Genotoxic endpoints in zebrafish stressed out sites on Morača and Lim rivers which are under pressures of fecal pollution. The data in this chapter provides an insight into current status obtained by the ex situ bioassays and indicates need for more comprehensive in situ assessment