Detection of SARS-CoV-2 RNA in the Danube River in Serbia associated with the discharge of untreated wastewaters.

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.

Indicators of Groundwater Potential for Nitrate Transformation in a Reductive Environment.

Increasing nitrate concentration in many shallow aquifers has become a global environmental problem. In order to determine anoxic aquifer sensitivity to nitrate pollution and potential for nitrogen conservation, we have qualified potential nitrate transformation pathways, by conducting field tests, tracer study, physicochemical and microbiological analysis of the groundwater. Combination of in situ measuring and laboratory analysis of physicochemical properties and selected biological activity reactivity tests (BARTTM), has enabled estimation of different pathways of NO3- reduction. During five days of nitrate enrichment experiment there was no observed increase in NO3- concentration, nor any observed decrease of TN (total nitrogen) in the groundwater, while NH4+ production rates increased (with the peak concentrations of 4.97 mg N/L) on the first day of the experiment. We assume that inhibited respiratory denitrification and enhanced DNRA (dissimilatory reduction of nitrate to ammonium) may preserve available nitrogen in a bioavailable form of ammonium.

Massive fish mortality and Cylindrospermopsis raciborskii bloom in Aleksandrovac Lake.

This paper presents a case study of a massive fish mortality during a Cylindrospermopsis raciborskii bloom in Aleksandrovac Lake, Serbia in mid-December 2012. According to a preliminary investigation of the samples taken on November 6 before the fish mortalities and to extended analyses of samples taken on November 15, no values of significant physicochemical parameters emerged to explain the cause(s) of the fish mortality. No industrial pollutants were apparent at this location, and results excluded the likelihood of bacterial infections. Even after freezing, the dissolved oxygen concentration in the water was sufficient for fish survival. High concentrations of chlorophyll a and phaeophytin occurred in the lake, and phytoplankton bloom samples were lethal in Artemia salina bioassays. A bloom of the cyanobacterium C. raciborskii was recorded during November. Although the A. salina bioassays indicated the presence of toxic compounds in the cyanobacterial cells, the cyanotoxins, microcystins, cylindrospermopsin and saxitoxin were not detected.