R.A. Bell

In-situ tryptophan-like fluorescence: A real-time indicator of faecal contamination in drinking water supplies.

Enteric pathogens are typically inferred from the presence of surrogate indicator organisms such as thermotolerant (faecal) coliforms (TTCs). The analysis of TTCs requires time-consuming incubation in suitable laboratories, which can limit sampling resolution, particularly during critical pollution events. Here, we demonstrate the use of in-situ fluorimeters targeting tryptophan-like compounds as a rapid, reagentless indicator of TTCs in groundwater-derived potable water supplies in Africa. A range of other common indicators of TTCs were also determined including nitrate, turbidity, and sanitary risk survey scores. Sampling was conducted during both the dry and wet seasons to investigate seasonality. Tryptophan-like fluorescence was the most effective predictor of both presence/absence and number of TTCs during both seasons. Seasonal changes in tryptophan-like fluorescence in deeper supplies suggest it is transported more efficiently through the aquifer than TTCs. Moreover, the perennial elevated concentrations in some wells suggest it is more resilient than TTCs in groundwater. Therefore tryptophan-like fluorescence could also be a better indicator of some smaller, more easily transported, and long-lived, pathogenic enteric viruses. These sensors have the potential to be included in real-time pollution alert systems for drinking water supplies throughout the world, as well as for mapping enteric pathogen risks in developing regions.

Tracing enteric pathogen contamination in sub-Saharan African groundwater

Quantitative PCR (qPCR) can rapidly screen for an array of faecally-derived bacteria, which can be employed as tracers to understand groundwater vulnerability to faecal contamination. A microbial DNA qPCR array was used to examine 45 bacterial targets, potentially relating to enteric pathogens, in 22 groundwater supplies beneath the city of Kabwe, Zambia in both the dry and subsequent wet season. Thermotolerant (faecal) coliforms, sanitary risks, and tryptophan-like fluorescence, an emerging real-time reagentless faecal indicator, were also concurrently investigated. There was evidence for the presence of enteric bacterial contamination, through the detection of species and group specific 16S rRNA gene fragments, in 72% of supplies where sufficient DNA was available for qPCR analysis. DNA from the opportunistic pathogen Citrobacter freundii was most prevalent (69% analysed samples), with Vibrio cholerae also perennially persistent in groundwater (41% analysed samples). DNA from other species such as Bifidobacterium longum and Arcobacter butzleri was more seasonally transient. Bacterial DNA markers were most common in shallow hand-dug wells in laterite/saprolite implicating rapid subsurface pathways and vulnerability to pollution at the surface. Boreholes into the underlying dolomites were also contaminated beneath the city highlighting that a laterite/saprolite overburden, as occurs across much of sub-Saharan aquifer, does not adequately protect underlying bedrock groundwater resources. Nevertheless, peri-urban boreholes all tested negative establishing there is limited subsurface lateral transport of enteric bacteria outside the city limits. Thermotolerant coliforms were present in 97% of sites contaminated with enteric bacterial DNA markers. Furthermore, tryptophan-like fluorescence was also demonstrated as an effective indicator and was in excess of 1.4μg/L in all contaminated sites.

The Influence of Flow and Bed Slope on Gas Transfer in Steep Streams and Their Implications for Evasion of CO2

The evasion of greenhouse gases (including CO2, CH4 and N2O) from streams and rivers to the atmosphere is an important process in global biogeochemical cycles, but our understanding of gas transfer in steep (> 10%) streams, and under varying flows is limited. We investigated gas transfer using combined tracer injections of SF6 and salt. We used a novel experimental design in which we compared four very steep (18.4-29.4%) and four moderately steep (3.7-7.6%) streams, and conducted tests in each stream under low flow conditions and during a high discharge event. Most dissolved gas evaded over short distances (~100 and ~200-400 m respectively), so accurate estimates of evasion fluxes will require sampling of dissolved gases at these scales to account for local sources. We calculated CO2 gas transfer coefficients (KCO2) and found statistically significant differences between larger KCO2 values for steeper (mean 0.465 min-1) streams compared to those with shallower slopes (mean 0.109 min-1). Variations in flow had an even greater influence. KCO2 was substantially larger under high (mean 0.497 min-1) compared to low flow conditions (mean 0.077 min-1). We developed a statistical model to predict KCO2 using values of streambed slope x discharge which accounted for 94 % of the variation. We show that two models using slope and velocity developed by Raymond et al. [2012] for streams and rivers with shallower slopes, also provide reasonable estimates of our CO2 gas transfer velocities (kCO2; m d-1). We developed a robust field protocol which could be applied in future studies.