Changes in Escherichia coli to enteric protozoa ratios in rivers: Implications for risk-based assessment of drinking water treatment requirements.

Minimum treatment requirements are set in response to established or anticipated levels of enteric pathogens in the source water of drinking water treatment plants (DWTPs). For surface water, contamination can be determined directly by monitoring reference pathogens or indirectly by measuring fecal indicators such as Escherichia coli (E. coli). In the latter case, a quantitative interpretation of E. coli for estimating reference pathogen concentrations could be used to define treatment requirements. This study presents the statistical analysis of paired E. coli and reference protozoa (Cryptosporidium, Giardia) data collected monthly for two years in source water from 27 DWTPs supplied by rivers in Canada. E. coli/Cryptosporidium and E. coli/Giardia ratios in source water were modeled as the ratio of two correlated lognormal variables. To evaluate the potential of E. coli for defining protozoa treatment requirements, risk-based critical mean protozoa concentrations in source water were determined with a reverse quantitative microbial risk assessment (QMRA) model. Model assumptions were selected to be consistent with the World Health Organization (WHO) Guidelines for drinking-water quality. The sensitivity of mean E. coli concentration trigger levels to identify these critical concentrations in source water was then evaluated. Results showed no proportionalities between the log of mean E. coli concentrations and the log of mean protozoa concentrations. E. coli/protozoa ratios at DWTPs supplied by small rivers in agricultural and forested areas were typically 1.0 to 2.0-log lower than at DWTPs supplied by large rivers in urban areas. The seasonal variations analysis revealed that these differences were related to low mean E. coli concentrations during winter in small rivers. To achieve the WHO target of 10-6 disability-adjusted life year (DALY) per person per year, a minimum reduction of 4.0-log of Cryptosporidium would be required for 20 DWTPs, and a minimum reduction of 4.0-log of Giardia would be needed for all DWTPs. A mean E. coli trigger level of 50 CFU 100 mL-1 would be a sensitive threshold to identify critical mean concentrations for Cryptosporidium but not for Giardia. Treatment requirements higher than 3.0-log would be needed at DWTPs with mean E. coli concentrations as low as 30 CFU 100 mL-1 for Cryptosporidium and 3 CFU 100 mL-1 for Giardia. Therefore, an E. coli trigger level would have limited value for defining health-based treatment requirements for protozoa at DWTPs supplied by small rivers in rural areas.

Microbacterium algeriense sp. nov., a novel actinobacterium isolated from Algerian oil production waters.

A non-motile, straight-rod-shaped, Gram-stain-positive and facultative anaerobic bacterium (i.e., strain G1T) was isolated from production waters from an Algerian oilfield. Growth was observed in the presence of 0.3-3.5 % (w/v) NaCl, at 20-50 °C and at pH 6.0-9.0. Results of phylogenetic analyses based on 16S rRNA gene sequences showed that strain G1T belonged to the genus Microbacterium. Strain G1 T was closely related to Microbacterium oxydans (DSM 20578T) and Microbacterium maritypicum (DSM 12512T) with 99.8 % sequence similarity and to Microbacterium saperdae (DSM 20169T) with 99.6 % sequence similarity. Strain G1 T contained MK9, MK10, MK11, MK12 and MK13 as respiratory quinones, and phosphatidylglycerol, diphosphatidylglycerol and glycolipid as the major polar lipids. The major cellular fatty acids were anteiso-C15:0, iso-C16:0 and anteiso-C17:0. The estimated DNA G+C content was 69.57 mol% based on its draft genome sequence. Genome annotation of strain G1T predicted the presence of 3511 genes, of which 3483 were protein-coding and 47 were tRNA genes. The DNA-DNA hybridization (DDH) and average nucleotide identity (ANI) values between strain G1T and M. oxydans (DSM 20578T) and M. maritypicum (DSM 12512T) were in both cases far below the respective species boundary thresholds (27.5 and 28.0 % for DDH; and 84.40 and 84.82% for ANI, respectively). Based on the data presented above, strain G1T was considered to represent a novel species for which the name Microbacterium algeriense is proposed with the type strain G1T (=DSM 109018T=LMG 31276T).

Near real-time notification of water quality impairments in recreational freshwaters using rapid online detection of ß-D-glucuronidase activity as a surrogate for Escherichia coli monitoring.

Waterborne disease outbreaks associated with recreational waters continue to be reported around the world despite existing microbiological water quality monitoring frameworks. Most regulations resort to the use of culture-based enumeration of faecal indicator bacteria such as Escherichia coli to protect bathers from gastrointestinal illness risks. However, the long sample-to-result time of standard culture-based assays (minimum 18-24 h) and infrequent regulatory sampling (weekly or less) do not enable detection of episodic water quality impairments and associated public health risks. The objective of this study was to assess the suitability of an autonomous online technology measuring ß-D-glucuronidase (GLUC) activity for near real-time monitoring of microbiological water quality in recreational waters and for the resulting beach management decisions. GLUC activity and E. coli concentrations were monitored at three freshwater sites in Quebec, Canada (sites Qc1-3) and one site in New Zealand (site NZ) between 2016 and 2018. We found site-dependent linear relationships between GLUC activity and E. coli concentrations and using confusion matrices, we developed site-specific GLUC activity beach action values (BAVs) matching the regulatory E. coli BAVs. Using the regulatory E. coli BAV as the gold standard, rates of false alarms (unnecessary beach advisories using GLUC activity BAV) and failures to act (failure to trigger advisories using GLUC activity) ranged between 0 and 32% and between 3 and 10%, respectively, which is comparable to the rates reported in other studies using qPCR-defined BAVs. However, a major benefit of the autonomous enzymatic technology is the real-time reporting of threshold exceedances, while temporal trends in GLUC activity can assist in understanding the underlying dynamics of faecal pollution and potential health risks. Our study is the first to describe the applicability of online near real-time monitoring of microbiological water quality as a tool for improved beach management and public health protection.

Tracking the contribution of multiple raw and treated wastewater discharges at an urban drinking water supply using near real-time monitoring of ß-d-glucuronidase activity.

Past waterborne outbreaks have demonstrated that informed vulnerability assessment of drinking water supplies is paramount for the provision of safe drinking water. Although current monitoring frameworks are not designed to account for short-term peak concentrations of fecal microorganisms in source waters, the recent development of online microbial monitoring technologies is expected to fill this knowledge gap. In this study, online near real-time monitoring of ß-d-glucuronidase (GLUC) activity was conducted for 1.5 years at an urban drinking water intake impacted by multiple point sources of fecal pollution. Parallel routine and event-based monitoring of E. coli and online measurement of physico-chemistry were performed at the intake and their dynamics compared over time. GLUC activity fluctuations ranged from seasonal to hourly time scales. All peak contamination episodes occurred between late fall and early spring following intense rainfall and/or snowmelt. In the absence of rainfall, recurrent daily fluctuations in GLUC activity and culturable E. coli were observed at the intake, a pattern otherwise ignored by regulatory monitoring. Cross-correlation analysis of time series retrieved from the drinking water intake and an upstream Water Resource Recovery Facility (WRRF) demonstrated a hydraulic connection between the two sites. Sewage by-passes from the same WRRF were the main drivers of intermittent GLUC activity and E. coli peaks at the drinking water intake following intense precipitation and/or snowmelt. Near real-time monitoring of fecal pollution through GLUC activity enabled a thorough characterization of the frequency, duration and amplitude of peak contamination periods at the urban drinking water intake while providing crucial information for the identification of the dominant upstream fecal pollution sources. To the best of our knowledge, this is the first characterization of a hydraulic connection between a WRRF and a downstream drinking water intake across hourly to seasonal timescales using high frequency microbial monitoring data. Ultimately, this should help improve source water protection through catchment mitigation actions, especially in a context of de facto wastewater reuse.

Autonomous online measurement of ß-D-glucuronidase activity in surface water: is it suitable for rapid E. coli monitoring?

Microbiological water quality is traditionally assessed using culture-based enumeration of faecal indicator bacteria such as Escherichia coli. Despite their relative ease of use, these methods require a minimal 18-24 h-incubation step before the results are obtained. This study aimed to assess the suitability of an autonomous online fluorescence-based technology measuring ß-glucuronidase (GLUC) activity for rapid near-real time monitoring of E. coli in water. The analytical precision was determined and compared to an automated microbial detection system, two culture-based assays and quantitative real-time PCR (qPCR). Using replicate measurements of grab samples containing E. coli concentrations between 50 and 2330 CFU.100 mL-1, the autonomous GLUC activity measurement technology displayed an average coefficient of variation (CV) of less than 5% that was 4-8-fold lower than other methods tested. Comparable precision was observed during online in situ monitoring of GLUC activity at a drinking water intake using three independent instruments. GLUC activity measurements were not affected by sewage or sediments at concentrations likely to be encountered during long-term monitoring. Furthermore, significant (p < 0.05) correlations were obtained between GLUC activity and the other assays including defined substrate technology (r = 0.77), membrane filtration (r = 0.73), qPCR (r = 0.55) and the automated microbial detection system (r = 0.50). This study is the first to thoroughly compare the analytical performance of rapid automated detection technologies to established culture and molecular-based methods. Results show that further research is required to correlate GLUC activity to the presence of viable E. coli as measured in terms of CFU.100 mL-1. This would allow the use of autonomous online GLUC activity measurements for rapid E. coli monitoring in water supplies used for drinking water production and recreation.

The Dynamic of a River Model Bacterial Community in Two Different Media Reveals a Divergent Succession and an Enhanced Growth of Most Strains Compared to Monocultures.

The dynamic of a community of 20 bacterial strains isolated from river water was followed in R2 broth and in autoclaved river water medium for 27 days in batch experiments. At an early stage of incubation, a fast-growing specialist strain, Acinetobater sp., dominated the community in both media. Later on, the community composition in both media diverged but was highly reproducible across replicates. In R2, several strains previously reported to degrade multiple simple carbon sources prevailed. In autoclaved river water, the community was more even and became dominated by several strains growing faster or exclusively in that medium. Those strains have been reported in the literature to degrade complex compounds. Their growth rate in the community was 1.5- to 7-fold greater than that observed in monoculture. Furthermore, those strains developed simultaneously in the community. Together, our results suggest the existence of cooperative interactions within the community incubated in autoclaved river water.

Occurrence and persistence of carbapenemases genes in hospital and wastewater treatment plants and propagation in the receiving river.

This study aims to investigate the prevalence of clinically relevant carbapenemases genes (blaKPC, blaNDM and blaOXA-48) in water samples collected over one-year period from hospital (H), raw and treated wastewater of two wastewater treatment plants (WWTPs) as well as along the Zenne River (Belgium). The genes were quantified in both particle-attached (PAB) and free-living (FLB) bacteria. Our results showed that absolute abundances were the highest in H waters. Although absolute abundances were significantly reduced in WWTP effluents, the relative abundance (normalized per 16S rRNA) was never lowered through wastewater treatment. Particularly, for the PAB the relative abundances were significantly higher in the effluents respect to the influents of both WWTPs for all the genes. The absolute abundances along the Zenne River increased from upstream to downstream, peaking after the release of WWTPs effluents, in both fractions. Our results demonstrated that blaKPC, blaNDM and blaOXA-48 are widely distributed in the Zenne as a consequence of chronic discharge from WWTPs. To conclude, the levels of carbapenemases genes are significantly lower than other genes conferring resistance to more widely used antibiotics (analyzed in previous studies carried out at the same sites), but could raise up to the levels of high prevalent resistance genes.

Antibiotic resistance along an urban river impacted by treated wastewaters.

Urban rivers are impacted ecosystems which may play an important role as reservoirs for antibiotic-resistant (AR) bacteria. The main objective of this study was to describe the prevalence of antibiotic resistance along a sewage-polluted urban river. Seven sites along the Zenne River (Belgium) were selected to study the prevalence of AR Escherichia coli and freshwater bacteria over a 1-year period. Culture-dependent methods were used to estimate E. coli and heterotrophic bacteria resistant to amoxicillin, sulfamethoxazole, nalidixic acid and tetracycline. The concentrations of these four antibiotics have been quantified in the studied river. The antibiotic resistance genes (ARGs), sul1, sul2, tetW, tetO, blaTEM and qnrS were also quantified in both particle-attached (PAB) and free-living (FLB) bacteria. Our results showed an effect of treated wastewaters release on the spread of antibiotic resistance along the river. Although an increase in the abundance of both AR E. coli and resistant heterotrophic bacteria was observed from upstream to downstream sites, the differences were only significant for AR E. coli. A significant positive regression was also found between AR E. coli and resistant heterotrophic bacteria. The concentration of ARGs increased from upstream to downstream sites for both particle-attached (PAB) and free-living bacteria (FLB). Particularly, a significant increase in the abundance of four among six ARGs analyzed was observed after crossing urban area. Although concentrations of tetracycline significantly correlated with tetracycline resistance genes, the antibiotic levels were likely too low to explain this correlation. The analysis of ARGs in different fractions revealed a significantly higher abundance in PAB compared to FLB for tetO and sul2 genes. This study demonstrated that urban activities may increase the spread of antibiotic resistance even in an already impacted river.

Application of a multidisciplinary and integrative weight-of-evidence approach to a 1-year monitoring survey of the Seine River.

Quality assessment of environments under high anthropogenic pressures such as the Seine Basin, subjected to complex and chronic inputs, can only be based on combined chemical and biological analyses. The present study integrates and summarizes a multidisciplinary dataset acquired throughout a 1-year monitoring survey conducted at three workshop sites along the Seine River (PIREN-Seine program), upstream and downstream of the Paris conurbation, during four seasonal campaigns using a weight-of-evidence approach. Sediment and water column chemical analyses, bioaccumulation levels and biomarker responses in caged gammarids, and laboratory (eco)toxicity bioassays were integrated into four lines of evidence (LOEs). Results from each LOE clearly reflected an anthropogenic gradient, with contamination levels and biological effects increasing from upstream to downstream of Paris, in good agreement with the variations in the structure and composition of bacterial communities from the water column. Based on annual average data, the global hazard was summarized as "moderate" at the upstream station and as "major" at the two downstream ones. Seasonal variability was also highlighted; the winter campaign was least impacted. The model was notably improved using previously established reference and threshold values from national-scale studies. It undoubtedly represents a powerful practical tool to facilitate the decision-making processes of environment managers within the framework of an environmental risk assessment strategy.

Spatiotemporal Dynamics of Total Viable Vibrio spp. in a NW Mediterranean Coastal Area.

A cellular approach combining Direct Viable Counting and Fluorescent In Situ Hybridization using a one-step multiple-probe technique and Solid Phase Cytometry (DVC-FISH-SPC) was developed to monitor total viable vibrios and cover the detection of a large diversity of vibrios. FISH combined three probes in the same assay and targeted sequences located at different positions on the 16S rRNA of Vibrio and Aliivibrio members. We performed a 10-month in situ study to investigate the weekly dynamics of viable vibrios relative to culturable counts at two northwestern Mediterranean coastal sites, and identified the key physicochemical factors for their occurrence in water using a multivariate analysis. Total viable and culturable cell counts showed the same temporal pattern during the warmer season, whereas the ratios between both methods were inverted during the colder seasons (<15°C), indicating that some of the vibrio community had entered into a viable but non-culturable (VBNC) state. We confirmed that Seawater Surface Temperature explained 51-62% of the total variance in culturable counts, and also showed that the occurrence of viable vibrios is controlled by two variables, pheopigment (15%) and phosphate (12%) concentrations, suggesting that other unidentified factors play a role in maintaining viability.

Carbon utilization profiles of river bacterial strains facing sole carbon sources suggest metabolic interactions.

Microbial communities play a key role in water self-purification. They are primary drivers of biogenic element cycles and ecosystem processes. However, these communities remain largely uncharacterized. In order to understand the diversity-heterotrophic activity relationship facing sole carbon sources, we assembled a synthetic community composed of 20 'typical' freshwater bacterial species mainly isolated from the Zenne River (Belgium). The carbon source utilization profiles of each individual strain and of the mixed community were measured in Biolog Phenotype MicroArrays PM1 and PM2A microplates that allowed testing 190 different carbon sources. Our results strongly suggest interactions occurring between our planktonic strains as our synthetic community showed metabolic properties that were not displayed by its single components. Finally, the catabolic performances of the synthetic community and a natural community from the same sampling site were compared. The synthetic community behaved like the natural one and was therefore representative of the latter in regard to carbon source consumption.

Change in the Structure of Escherichia coli Population and the Pattern of Virulence Genes along a Rural Aquatic Continuum.

The aim of this study was to investigate the diversity of the Escherichia coli population, focusing on the occurrence of pathogenic E. coli, in surface water draining a rural catchment. Two sampling campaigns were carried out in similar hydrological conditions (wet period, low flow) along a river continuum, characterized by two opposite density gradients of animals (cattle and wild animals) and human populations. While the abundance of E. coli slightly increased along the river continuum, the abundance of both human and ruminant-associated Bacteroidales markers, as well as the number of E. coli multi-resistant to antibiotics, evidenced a fecal contamination originating from animals at upstream rural sites, and from humans at downstream urban sites. A strong spatial modification of the structure of the E. coli population was observed. At the upstream site close to a forest, a higher abundance of the B2 phylogroup and Escherichia clade strains were observed. At the pasture upstream site, a greater proportion of both E and B1 phylogroups was detected, therefore suggesting a fecal contamination of mainly bovine origin. Conversely, in downstream urban sites, A, D, and F phylogroups were more abundant. To assess the occurrence of intestinal pathogenic strains, virulence factors [afaD, stx1, stx2, eltB (LT), estA (ST), ipaH, bfpA, eae, aaiC and aatA] were screened among 651 E. coli isolates. Intestinal pathogenic strains STEC O174:H21 (stx2) and EHEC O26:H11 (eae, stx1) were isolated in water and sediments close to the pasture site. In contrast, in the downstream urban site aEPEC/EAEC and DAEC of human origin, as well as extra-intestinal pathogenic E. coli belonging to clonal group A of D phylogroup, were sampled. Even if the estimated input of STEC (Shiga toxin-producing E. coli) - released in water at the upstream pasture site - at the downstream site was low, we show that STEC could persist in sediment. These results show that, the run-off of small cattle farms contributed, as much as the wastewater effluent, in the dissemination of pathogenic E. coli in both water and sediments, even if the microbiological quality of the water was good or to average quality according to the French water index.

How does the cladoceran Daphnia pulex affect the fate of Escherichia coli in water?

The faecal indicator Escherichia coli plays a central role in water quality assessment and monitoring. It is therefore essential to understand its fate under various environmental constraints such as predation by bacterivorous zooplankton. Whereas most studies have examined how protozooplankton communities (heterotrophic nanoflagellates and ciliates) affect the fate of E. coli in water, the capacity of metazooplankton to control the faecal indicator remains poorly understood. In this study, we investigated how the common filter-feeding cladoceran, Daphnia pulex, affects the fate of E. coli under different experimental conditions. Daphnia ingested E. coli and increased its loss rates in water, but the latter rates decreased from 1.65 d-1 to 0.62 d-1 after a 1,000-fold reduction in E. coli initial concentrations, due to lower probability of encounter between Daphnia and E. coli. The combined use of culture and PMA qPCR (viability-qPCR) demonstrated that exposure to Daphnia did not result into the formation of viable but non-culturable E. coli cells. In lake water, a significant part of E. coli population loss was associated with matrix-related factors, most likely due to predation by other bacterivorous biota and/or bacterial competition. However, when exposing E. coli to a D. pulex gradient (from 0 to 65 ind.L-1), we observed an increasing impact of Daphnia on E. coli loss rates, which reached 0.47 d-1 in presence of 65 ind.L-1. Our results suggest that the filter-feeder can exert a non-negligible predation pressure on E. coli, especially during seasonal Daphnia population peaks. Similar trials using other Daphnia species as well as stressed E. coli cells will increase our knowledge on the capacity of this widespread zooplankter to control E. coli in freshwater resources. Based on our results, we strongly advocate the use of natural matrices to study these biotic interactions in order to avoid overestimation of Daphnia impact.

Modelling of faecal indicator bacteria (FIB) in the Red River basin (Vietnam).

Many studies have been published on the use of models to assess water quality through faecal contamination levels. However, the vast majority of this work has been conducted in developed countries and similar studies from developing countries in tropical regions are lacking. Here, we used the Seneque/Riverstrahler model to investigate the dynamics and seasonal distribution of total coliforms (TC), an indicator of faecal contamination, in the Red River (Northern Vietnam) and its upstream tributaries. The results of the model showed that, in general, the overall correlations between the simulated and observed values of TC follow a 1:1 relationship at all examined stations. They also showed that TC numbers were affected by both land use in terms of human and livestock populations and by hydrology (river discharge). We also developed a possible scenario based on the predicted changes in future demographics and land use in the Red River system for the 2050 horizon. Interestingly, the results showed only a limited increase of TC numbers compared with the present situation at all stations, especially in the upstream Vu Quang station and in the urban Ha Noi station. This is probably due to the dominance of diffuse sources of contamination relative to point sources. The model is to our knowledge one of the first mechanistic models able to simulate spatial and seasonal variations of microbial contamination (TC numbers) in the whole drainage network of a large regional river basin covering both urban and rural areas of a developing country.

Cumulative effects of fecal contamination from combined sewer overflows: Management for source water protection.

The quality of a drinking water source depends largely on upstream contaminant discharges. Sewer overflows can have a large influence on downstream drinking water intakes as they discharge untreated or partially treated wastewaters that may be contaminated with pathogens. This study focuses on the quantification of Escherichia coli discharges from combined sewer overflows (CSOs) and the dispersion and diffusion in receiving waters in order to prioritize actions for source water protection. E. coli concentrations from CSOs were estimated from monitoring data at a series of overflow structures and then applied to the 42 active overflow structures between 2009 and 2012 using a simple relationship based upon the population within the drainage network. From these estimates, a transport-dispersion model was calibrated with data from a monitoring program from both overflow structures and downstream drinking water intakes. The model was validated with 15 extreme events such as a large number of overflows (n > 8) or high concentrations at drinking water intakes. Model results demonstrated the importance of the cumulative effects of CSOs on the degradation of water quality downstream. However, permits are typically issued on a discharge point basis and do not consider cumulative effects. Source water protection plans must consider the cumulative effects of discharges and their concentrations because the simultaneous discharge of multiple overflows can lead to elevated E. coli concentrations at a drinking water intake. In addition, some CSOs have a disproportionate impact on peak concentrations at drinking water intakes. As such, it is recommended that the management of CSOs move away from frequency based permitting at the discharge point to focus on the development of comprehensive strategies to reduce cumulative and peak discharges from CSOs upstream of drinking water intakes.

The effects of combined sewer overflow events on riverine sources of drinking water.

This study was set out to investigate the impacts of Combined Sewer Overflows (CSOs) on the microbiological water quality of a river used as a source of drinking water treatment plants. Escherichia coli concentrations were monitored at various stations of a river segment located in the Greater Montreal Area including two Drinking Water Intakes (DWIs) in different weather conditions (dry weather and wet weather (precipitation and snowmelt period)). Long-term monitoring data (2002-2011) at DWIs revealed good microbiological water quality with E. coli median concentrations of 20 and 30 CFU/100 mL for DWI-1 and DWI-2 respectively. However, E. coli concentration peaks reached up to 510 and 1000 CFU/100 mL for both DWIs respectively. Statistical Process Control (SPC) analysis allowed the identification of E. coli concentration peaks in almost a decade of routine monitoring data at DWIs. Almost 80% of these concentrations were linked to CSO discharges caused by precipitation exceeding 10 mm or spring snowmelt. Dry weather monitoring confirmed good microbiological water quality. Wet weather monitoring showed an increase of approximately 1.5 log of E. coli concentrations at DWIs. Cumulative impacts of CSO discharges were quantified at the river center with an increase of approximately 0.5 log of E. coli concentrations. Caffeine (CAF) was tested as a potential chemical indicator of CSO discharges in the river and CAF concentrations fell within the range of previous measurements performed for surface waters in the same area (∼20 ng/L). However, no significant differences were observed between CAF concentrations in dry and wet weather, as the dilution potential of the river was too high. CSO event based monitoring demonstrated that current bi-monthly or weekly compliance monitoring at DWIs underestimate E. coli concentrations entering DWIs and thus, should not be used to quantify the risk at DWIs. High frequency event-based monitoring is a desirable approach to establish the importance and duration of E. coli peak concentrations entering DWIs.

Pelagic photoferrotrophy and iron cycling in a modern ferruginous basin.

Iron-rich (ferruginous) ocean chemistry prevailed throughout most of Earth's early history. Before the evolution and proliferation of oxygenic photosynthesis, biological production in the ferruginous oceans was likely driven by photoferrotrophic bacteria that oxidize ferrous iron {Fe(II)} to harness energy from sunlight, and fix inorganic carbon into biomass. Photoferrotrophs may thus have fuelled Earth's early biosphere providing energy to drive microbial growth and evolution over billions of years. Yet, photoferrotrophic activity has remained largely elusive on the modern Earth, leaving models for early biological production untested and imperative ecological context for the evolution of life missing. Here, we show that an active community of pelagic photoferrotrophs comprises up to 30% of the total microbial community in illuminated ferruginous waters of Kabuno Bay (KB), East Africa (DR Congo). These photoferrotrophs produce oxidized iron {Fe(III)} and biomass, and support a diverse pelagic microbial community including heterotrophic Fe(III)-reducers, sulfate reducers, fermenters and methanogens. At modest light levels, rates of photoferrotrophy in KB exceed those predicted for early Earth primary production, and are sufficient to generate Earth's largest sedimentary iron ore deposits. Fe cycling, however, is efficient, and complex microbial community interactions likely regulate Fe(III) and organic matter export from the photic zone.

Vertical Distribution of Functional Potential and Active Microbial Communities in Meromictic Lake Kivu.

The microbial community composition in meromictic Lake Kivu, with one of the largest CH4 reservoirs, was studied using 16S rDNA and ribosomal RNA (rRNA) pyrosequencing during the dry and rainy seasons. Highly abundant taxa were shared in a high percentage between bulk (DNA-based) and active (RNA-based) bacterial communities, whereas a high proportion of rare species was detected only in either an active or bulk community, indicating the existence of a potentially active rare biosphere and the possible underestimation of diversity detected when using only one nucleic acid pool. Most taxa identified as generalists were abundant, and those identified as specialists were more likely to be rare in the bulk community. The overall number of environmental parameters that could explain the variation was higher for abundant taxa in comparison to rare taxa. Clustering analysis based on operational taxonomic units (OTUs at 0.03 cutoff) level revealed significant and systematic microbial community composition shifts with depth. In the oxic zone, Actinobacteria were found highly dominant in the bulk community but not in the metabolically active community. In the oxic-anoxic transition zone, highly abundant potentially active Nitrospira and Methylococcales were observed. The co-occurrence of potentially active sulfur-oxidizing and sulfate-reducing bacteria in the anoxic zone may suggest the presence of an active yet cryptic sulfur cycle.

Assessing the impacts of wastewater treatment implementation on the water quality of a small urban river over the past 40 years.

This paper studies the effects of the implementation of wastewater treatment (WWT) on the water quality of small urban river systems by considering as an extreme case study (volumetric contribution of wastewaters >50%) the evolution of the Zenne River waters (Belgium) over the last 40 years. In urban rivers, organic matter (OM), oxygen, and nutrients are primarily controlled by wastewater releases which depend on the population and the WWT capacity in the river basin, the latter being dependent on environmental policy decisions. We introduce a novel basin-scale evaluation method that considers the evolution of annual pollutant loads at the outlet of the river basin directly as a function of WWT capacity. Based on this approach, we could prove that the load reductions observed after the implementation of WWT in the river basin was a good indicator of the global treatment efficiency of the WWT plants. We also show that high self-purification processes within the river basin may lead to reach minimum levels of OM before the completion of WWT. In addition, the effects of wet weather conditions did also change as a function of the WWT capacity going from positive effects at low capacity to negative effects at high capacity. Finally, the full implementation of WWT in urban river basins does not necessarily guarantee a good status for water quality, mostly because of the high volumetric proportion of treated wastewaters, which do not have the quality standards of river waters.

Temporal analysis of E. coli, TSS and wastewater micropollutant loads from combined sewer overflows: implications for management.

A combined sewer overflow (CSO) outfall was monitored to assess the impact of temporal mass loads on the appropriateness of treatment options. Instantaneous loads (mass per s) varied by approximately three orders of magnitude during events (n = 9 in spring, summer and the fall) with no significant seasonal variations. The median fraction of total loads discharged with the first 25% of the total volume ranged from 28% (theophylline) to 40% (Total Suspended Solids (TSS)) and loads remained high for the duration of the events. E. coli and TSS loads originated primarily from wastewater (WW) (63% and 75%, respectively). However, a mix of stormwater (SW) and sewer deposit (SD) resuspension contributed from 73 to 95% for the first 50% of the volume discharged of total TSS loads for 2 events. The contribution of SD resuspension was not negligible for Wastewater Micropollutants (WWMPs), especially for carbamazepine. Sustained high loads over the course of CSOs highlight the need to revisit current CSO and SW management strategies that focus on the treatment of early discharge volumes.