Multiple fluorescence approaches to identify rapid changes in microbial indicators at karst springs

Karst springs are globally important for drinking water supply but are often also exceptionally vulnerable to contamination. Such springs usually exhibit strong variation in microbial water quality in sharp response to rainfall events, thus, posing a health hazard to consumers of water supplied from these sources. The rapid detection of such changes is extremely important as well as being able to establish a link to the sources of such pollution, so that appropriate measures can be taken both in terms of immediate protection of human health and the management of karst aquifers. In this study, a fluorescence-based multi-parameter approach was trialed in order to evaluate which methods can be used to monitor rainfall-induced rapid changes in microbial water quality at karst springs, as well as determine whether such changes can be linked to sources of human effluent contamination. The results from three monitoring periods at two karst springs revealed marked responses to rainfall events for all of the microbial parameters measured. Total cell count (TCC) measurements using flow cytometry (FCM) showed very strong positive correlations with the more conventionally monitored faecal indicator bacteria (FIB) and total coliforms (TC), indicating that such a fluorescence-based and cultivation-independent technique can be very useful to indicate rapid changes in microbial water quality at karst springs. Furthermore, very strong positive correlations were also found between tryptophan-like fluorescence (TLF) measurements and concentrations of all monitored microbial parameters, again demonstrating that such a fluorescence-based approach can also be useful for detecting rapid changes in concentrations of traditional faecal indicators. Interestingly, it was found that fluorescent whitening compounds (FWCs) signals do not necessarily follow temporal variations of microbial indicators. However, the frequency of detection of positive FWCs signals may still reveal useful information about the overall magnitude of human wastewater effluent impacts on karst aquifer systems.