Integrating molecular microbial methods to improve faecal pollution management in rivers with designated bathing waters.

Rivers are at risk from a variety of pollution sources. Faecal pollution is of particular concern since it disperses pathogenic microorganisms in the aquatic environment. Currently, faecal pollution levels in rivers is monitored using faecal indicator bacteria (FIB) that do not offer information about pollution sources and associated risks. This study used a combined molecular approach, along with measurements of water quality, to gain information on pollution sources, and risk levels, in a newly designated recreational bathing site in the River Wharfe (UK). Physico-chemical parameters were monitored in situ, with water quality multiparameter monitoring sondes installed during the 2021 bathing season. The molecular approach was based on quantitative PCR (qPCR)-aided Microbial Source Tracking (MST) and 16S rRNA gene metabarcoding to obtain a fingerprint of bacterial communities and identify potential bioindicators. The analysis from the water quality sondes showed that ammonium was the main parameter determining the distribution of FIB values. Lower faecal pollution levels were detected in the main river when compared to tributaries, except for samples in the river located downstream of a wastewater treatment plant. The faecal pollution type (anthropogenic vs. zoogenic) changed the diversity and the structure of bacterial communities, giving a distinctive fingerprint that can be used to inform source. DNA-based methods showed that the presence of human-derived bacteria was associated with Escherichia coli spikes, coinciding with higher bacterial diversity and the presence of potential pathogenic bacteria mainly of the genus Mycobacterium, Aeromonas and Clostridium. Samples collected after a heavy rainfall event were associated with an increase in Bacteroidales, which are markers of faecal pollution, including Bacteroides graminisolvens, a ruminant marker associated with surface run-off from agricultural sources. The combined use of qPCR and 16S rRNA sequencing was able to identify pollution sources, and novel bacterial indicators, thereby aiding decision-making and management strategies in recreational bathing rivers.

Altitudinal gradients of PBDEs and PCBs in fish from European high mountain lakes.

A first case of temperature-dependent distribution of polybromodiphenyl ethers (PBDEs) in remote areas is shown. Analysis of these compounds in fish from Pyrenean lakes distributed along an altitudinal transect shows higher concentrations at lower temperatures, as predicted in the global distillation model. Conversely, no temperature-dependent distribution is observed in a similar transect in the Tatra mountains (Central Europe) nor in fish from high mountain lakes distributed throughout Europe. The fish concentrations of polychlorobiphenyls (PCBs) examined for comparison showed significant temperature correlations in all these studied lakes. Cold trapping of both PCBs and PBDEs concerned the less volatile congeners. In the Pyrenean lake transect the concentrations of PCBs and PBDEs in fish were correlated despite the distinct use of these compounds and their 40 year time lag of emissions to the environment. Thus, temperature effects have overcome these anthropogenic differences constituting at present the main process determining their distributions. These cases of distinct PBDEs and PCBs behavior in high mountains likely reflect early stages in the environmental distribution of the former since they have been under secondary redistribution processes over much shorter time than the latter.

Influence of altitude and age in the accumulation of organochlorine compounds in fish from high mountain lakes.

The analysis of hexachlorobenzene, hexachlorocyclohexanes, polychlorobiphenyls, and DDTs in muscle of fish from high mountain lakes shows that a proportion of their concentration variance depends on fish age and lake altitude. Interestingly, the magnitude of this share corresponds linearly with the log-transformed vapor pressure (Vp) of the organochlorine compounds (OC). Thus, the distributions of OC with Vp < 10(-2.5) Pa are mostly determined by these two variables. Altitude gradients mainly respond to temperature differences, involving concentration increases of 25-150 times between 8.7 and -2.3 degrees C. The age effect encompasses concentration increments of 2.4-7.8 for average lake differences between 2 and 13 yr. However, both effects are independent since no correlation between fish age and lake altitude is observed. Fish liver concentrations exhibit the same pattern, but the correlations are only significantfor age, suggesting thatthe temperature trend is more related to long-term accumulation than episodic intake. The temperature effect is independent from compound origin. In addition, the sites situated at highest altitude, those most distant from possible ground pollution sources, are the most polluted. The results can be explained by condensation effects such as those described for the latitudinal trends that support the global distillation theory. However, in the high altitude lakes a temperature-dependent amplification mechanism, probably related to low metabolism and respiration at lowtemperatures, enhances OC accumulation in fish beyond the increases predicted from theoretical condensation and solubilization enthalpies. The observed temperature dependence suggests that a general remobilization of OC accumulated in high mountain areas could take place as a consequence of the general warming of these areas anticipated in the climatic change studies.