Sedimentary records of sewage pollution using faecal markers in contrasting peri-urban shallow lakes

Sewage contamination in shallow lake sediments is of concern because the pathogens, organic matter and nutrients contribute to the deterioration of the water-bodies' health and ecology. Sediment cores from three shallow lakes (Coneries, Church and Clifton Ponds) within Attenborough nature reserve located downstream of sewage treatment works were analysed for TOC, C/N, δ¹³C, δ¹⁵N, bacterial coliforms and faecal sterols. ²¹⁰Pb and ¹³⁷Cs activities were used to date the sediments. Elemental analysis suggests that the source of organic matter was algal and down profile changes in δ¹³C indicate a possible decrease in productivity with time which could be due to improvements in sewage treatment. δ¹⁵N for Coneries Pond are slightly higher than those observed in Church or Clifton and are consistent with a sewage-derived nitrate source which has been diluted by non-sewage sources of N. The similarity in δ¹⁵N values (+ 12‰ to + 10‰) indicates that the three ponds were not entirely hydrologically isolated. Analysis by gas chromatography/mass spectrometry (GC/MS) reveals that Coneries Pond had sterol concentrations in the range 20 to 30 μg/g (dry wt.), whereas, those from Clifton and Church Ponds were lower. The highest concentrations of the human-sourced sewage marker 5β-coprostanol were observed in the top 40 cm of Coneries Pond with values up to 2.2 μg/g. In contrast, Church and Clifton Pond sediments contain only trace amounts throughout. Down-profile comparison of 5β-coprostanol/cholesterol, 5β-coprostanol/(5β-coprostanol + 5α-cholestanol) and 5β-epicoprostanol/coprostanol as well as 5α-cholestanol/cholesterol suggests that Coneries Pond has received appreciable amounts of faecal contamination. Examination of 5β-stigmastanol (marker for herbivorous/ruminant animals) down core concentrations suggests a recent decrease in manure slurry input to Coneries Pond. The greater concentration of β-sitosterol in sediments from Church and Clifton Ponds as compared to Coneries is attributed in part to their greater diversity and extent of aquatic plants and avian faeces.     

Chemometric approach to validating faecal sterols as source tracer for faecal contamination in water

Faecal sterols detection is a promising method for identifying sources of faecal pollution. In this study, faecal contamination in water samples from point source (sewage treatment plants, chicken farms, quail farms and horse stables) was extracted using the solid phase extraction (SPE) technique. Faecal sterols (coprostanol, cholesterol, stigmasterol, β-sitosterol and stigmastanol) were selected as parameters to differentiate the source of faecal pollution. The results indicated that coprostanol, cholesterol and β-sitosterol were the most significant parameters that can be used as source tracers for faecal contamination. Chemometric techniques, such as cluster analysis, principal component analysis and discriminant analysis were applied to the data set on faecal contamination in water from various pollution sources in order to validate the faecal sterols’ profiles. Cluster analysis generated three clusters: coprostanol was in cluster 1, cholesterol and β-sitosterol formed cluster 2, while cluster 3 contained stigmasterol and stigmastanol. Discriminant analysis suggested that coprostanol, cholesterol and β-sitosterol were the most significant parameters to discriminate between the faecal pollution source. The use of chemometric techniques provides useful and promising indicators in tracing the source of faecal contamination.     

Baseline isotope data for Spirodela sp.: nutrient differentiation in aquatic systems

The excessive addition of nitrogen to watersheds is recognized as one of the main causes of the global deterioration of aquatic ecosystems and an increasing number of studies have shown that δ¹⁵N signatures of macrophytes may reflect the N-loading of the system under investigation. This study investigated isotopic equilibration rates and concentration level effects of KNO₃ and cow manure nutrient solutions on the δ¹⁵N and δ¹³C signatures, C/N ratios, % N and % C of Spirodela sp. over time, to determine the feasibility of their use in monitoring anthropogenic N-loading in freshwater systems. Spirodela δ¹⁵N signatures clearly distinguished between nutrient types within 2 days of introduction, with plants grown in KNO₃ showing extremely depleted δ¹⁵N values (−15.00 to −12.00‰) compared to those growing in cow manure (14.00-18.00‰). Isotopic equilibration rates could not be determined with certainty, but plant isotopic differentiation between nutrient regimes became apparent after 2 days and started to equilibrate by day 4. Concentration level effects were also apparent, with Spirodela tissue displaying more depleted and enriched δ¹⁵N values in higher concentrations of KNO₃ and cow manure respectively. δ¹³C signatures of some plants grown in manure were more enriched than plants grown in KNO₃ and reverse osmosis (RO) water. However, nutrient induced differences in δ¹³C were small and are likely to be obscured in the natural environment. Decreased C/N ratios and increases in plant % N in zero N concentration treatments confirmed the presence of a commensal cyanobacterial-duckweed association within Spirodela sp., reducing its effectiveness as an in-situ incubator in low nutrient environments. However, indications are that Spirodela may make a useful isotope monitoring tool under conditions of long-term, continuous nutrient inputs such as systems impacted by sewage outfalls and/or wastewater inputs.     

Molecular subtypes of Campylobacter spp., Salmonella enterica, and Escherichia coli O157:H7 isolated from faecal and surface water samples in the Oldman River watershed, Alberta, Canada

Campylobacter spp., Salmonella enterica, and Escherichia coli O157:H7 isolated from 898 faecal, 43 sewage, and 342 surface water samples from the Oldman River were characterized using bacterial subtyping methods in order to investigate potential sources of contamination of the watershed. Among these pathogens, Campylobacter spp. were the most frequently isolated from faecal, sewage, and surface water samples (266/895, 11/43, and 91/342, respectively), followed by Salmonella (67/898, 8/43, and 29/342, respectively), and E. coli O157:H7 (16/898, 2/43, and 8/342, respectively). Salmonella Rubislaw was the most common serovar isolated from water. This serovar was also isolated from two wild bird species. Most other serovars isolated from water were either not isolated from animals or were isolated from multiple species. E. coli O157:H7 was predominantly isolated from cattle. The most common phage-types of this pathogen from cattle were also the most common among water isolates, and there were exact pulsed field gel electrophoresis and comparative genomic fingerprint matches between cattle, sewage, and water isolates. Campylobacters were commonly isolated from surface waters and faeces from most animal species. Restriction fragment length polymorphism of the Campylobacter flaA gene identified several location and host species-specific (cattle, goose, pig) fingerprints. Molecular subtyping of these bacterial pathogens shows considerable promise as a tool for determining the sources of faecal pollution of water.     

Evaluation of wastewater nitrogen transformation in a natural wetland (Ulaanbaatar, Mongolia) using dual-isotope analysis of nitrate

The Tuul River, which provides water for the daily needs of many residents of Ulaanbaatar, Mongolia, has been increasingly polluted by wastewater from the city's sewage treatment plant. Information on water movement and the transformation of water-borne materials is required to alleviate the deterioration of water quality. We conducted a synoptic survey of general water movement, water quality including inorganic nitrogen concentrations, and isotopic composition of nitrogen (δ¹⁵N-NO₃⁻, δ¹⁸O-NO₃⁻, and δ¹⁵N-NH₄⁺) and water (δ¹⁸O-H₂O) in a wetland area that receives wastewater before it enters the Tuul River. We sampled surface water, groundwater, and spring water along the two major water routes in the wetland that flow from the drain of the sewage treatment plant to the Tuul River: a continuous tributary and a discontinuous tributary. The continuous tributary had high ammonium (NH₄⁺) concentrations and nearly stable δ¹⁵N-NH₄⁺, δ¹⁵N-NO₃⁻, and δ¹⁸O-NO₃⁻ concentrations throughout its length, indicating that nitrogen transformation (i.e., nitrification and denitrification) during transit was small. In contrast, NH₄⁺ concentrations decreased along the discontinuous tributary and nitrate (NO₃⁻) concentrations were low at many points. Values of δ¹⁵N-NH₄⁺, δ¹⁵N-NO₃⁻, and δ¹⁸O-NO₃⁻ increased with flow along the discontinuous route. Our results indicate that nitrification and denitrification contribute to nitrogen removal in the wetland area along the discontinuous tributary with slow water transport. Differences in hydrological pathways and the velocity of wastewater transport through the wetland area greatly affect the extent of nitrogen removal.     

Carbon cycling in a zero-discharge mariculture system

Interest in mariculture systems will rise in the near future due to the decreased ability of the ocean to supply the increasing demand for seafood. We present a trace study using stable carbon and nitrogen isotopes and chemical profiles of a zero-discharge mariculture system stocked with the gilthead seabream (Sparus aurata). Water quality maintenance in the system is based on two biofiltration steps. Firstly, an aerobic treatment step comprising a trickling filter in which ammonia is oxidized to nitrate. Secondly, an anaerobic step comprised of a digestion basin and a fluidized bed reactor where excess organic matter and nitrate are removed. Dissolved inorganic carbon and alkalinity values were higher in the anaerobic loop than in the aerobic loop, in agreement with the main biological processes taking place in the two treatment steps. The δ¹³C of the dissolved inorganic carbon (δ¹³C_DIC) was depleted in ¹³C in the anaerobic loop as compared to the aerobic loop by 2.5-3‰. This is in agreement with the higher dissolved inorganic carbon concentrations in the anaerobic loop and the low water retention time and the chemolithotrophic activity of the aerobic loop. The δ¹³C and δ¹⁵N of organic matter in the mariculture system indicated that fish fed solely on feed pellets. Compared to feed pellets and particulate organic matter, the sludge in the digestion basin was enriched in ¹⁵N while δ¹³C was not significantly different. This latter finding points to an intensive microbial degradation of the organic matter taking place in the anaerobic treatment step of the system.     

Erratum to “Sources of nitrate and ammonium contamination in groundwater under developing Asian megacities”

The status of nitrate (NO₃⁻), nitrite (NO₂⁻) and ammonium (NH₄⁺) contamination in the water systems, and the mechanisms controlling their sources, pathways, and distributions were investigated for the Southeast Asian cities of Metro Manila, Bangkok, and Jakarta. GIS-based monitoring and dual isotope approach (nitrate δ¹⁵N and δ¹⁸O) suggested that human waste via severe sewer leakage was the major source of nutrient contaminants in Metro Manila and Jakarta urban areas. Furthermore, the characteristics of the nutrient contamination differed depending on the agricultural land use pattern in the suburban areas: high nitrate contamination was observed in Jakarta (dry fields), and relatively lower nutrients consisting mainly of ammonium were detected in Bangkok (paddy fields).The exponential increase in NO₃⁻-δ¹⁵N along with the NO₃⁻ reduction and clear δ¹⁸O/δ¹⁵N slopes of NO₃⁻ (∼ 0.5) indicated the occurrence of denitrification. An anoxic subsurface system associated with the natural geological setting (e.g., the old tidal plain at Bangkok) and artificial pavement coverage served to buffer NO₃⁻ contamination via active denitrification and reduced nitrification.Our results showed that NO₃⁻ and NH₄⁺ contamination of the aquifers in Metro Manila, Bangkok, and Jakarta was not excessive, suggesting low risk of drinking groundwater to human health, at present. However, the increased nitrogen load and increased per capita gross domestic product (GDP) in these developing cities may increase this contamination in the very near future. Continuous monitoring and management of the groundwater system is needed to minimize groundwater pollution in these areas, and this information should be shared among adjacent countries with similar geographic and cultural settings.     

Tracing the sources of nitrate in the Han River watershed in Korea, using delta15N-NO3- and delta18O-NO3- values

The dissolved nitrate concentrations and their nitrogen and oxygen isotopic ratios were analyzed in seasonal samples from Korea's Han River to ascertain the seasonal and spatial variations of dissolved nitrate and its possible sources. Nitrate concentrations in the South Han River (SHR) were much higher than those in the North Han River (NHR), probably because of the more extensive distribution of agricultural fields, residential areas and animal farms in the SHR drainage basin. The nitrogen isotopic composition of dissolved nitrate indicates that nitrate-nitrogen (NO(3)(-)-N) is derived mainly from atmospheric deposition and/or soil organic matter in the NHR but comes principally from manure or sewage, with only a minor contribution from atmospheric deposition or soil organic matter, in the SHR. The oxygen isotopic compositions of dissolved nitrate suggest that most atmospheric nitrate undergoes microbial nitrification before entering the river.     

Comparing isotopic groundwater age measurements with simulated groundwater ages: example of the Abbotsford–Sumas Aquifer (USA and Canada) and application

Elevated nitrate concentrations have been documented since the 1970s within the trans‐national Abbotsford–Sumas aquifer, situated in the central Fraser Valley of southern British Columbia, Canada, and northern Washington State, United States. Nitrate concentrations in excess of 10 mg/L NO₃–N are commonly observed in the monitoring wells. A groundwater model is used in this study to conduct a particle tracking analysis to estimate groundwater travel times and compare these with groundwater ages determined from ³H/³He concentrations measured in several monitoring wells. Groundwater ages estimated from particle tracking show excellent agreement with measured ages (slope=1), although there is scatter (R²=0.76), reflecting the complexity of the travel pathways and ambiguities in groundwater ages because of mixing. An assessment of the efficacy of recommended nutrient management practices, which were implemented in 1992 to reduce nitrogen loading, suggests that wells shallower than about 20 m should be monitoring a decrease in nitrate concentration. However, persistent elevated nitrate concentrations even at shallow depths point to the lingering effects of a remnant manure–nitrate signature in combination with continued high loading.     

Kinetics of nitrate and perchlorate reduction in ion-exchange brine using the membrane biofilm reactor (MBfR)

Several sources of bacterial inocula were tested for their ability to reduce nitrate and perchlorate in synthetic ion-exchange spent brine (30-45 g/L) using a hydrogen-based membrane biofilm reactor (MBfR). Nitrate and perchlorate removal fluxes reached as high as 5.4 g N m⁻² d⁻¹ and 5.0 g ClO₄ m⁻² d⁻¹, respectively, and these values are similar to values obtained with freshwater MBfRs. Nitrate and perchlorate removal fluxes decreased with increasing salinity. The nitrate fluxes were roughly first order in H₂ pressure, but roughly zero-order with nitrate concentration. Perchlorate reduction rates were higher with lower nitrate loadings, compared to high nitrate loadings; this is a sign of competition for H₂. Nitrate and perchlorate reduction rates depended strongly on the inoculum. An inoculum that was well acclimated (years) to nitrate and perchlorate gave markedly faster removal kinetics than cultures that were acclimated for only a few months. These results underscore that the most successful MBfR bioreduction of nitrate and perchlorate in ion-exchange brine demands a well-acclimated inoculum and sufficient hydrogen availability.     

Persistence of host-associated Bacteroidales gene markers and their quantitative detection in an urban and agricultural mixed prairie watershed

Microbial source tracking is an emerging tool developed to protect water sources from faecal pollution. In this study, we evaluated the suitability of real time-quantitative PCR (qPCR) Taqman assays developed for detection of host-associated Bacteroidales markers in a prairie watershed. The qPCR primers and probes used in this study exhibited high accuracy (88-96% sensitivity and ≥99% host specificity) in detecting Bacteroidales spp. that are associated with faeces from humans, ruminants, bovines, and horses. The ruminant- and human-associated markers were also found in high concentrations within individual faecal samples, ranging from 3.4 to 7.3 log₁₀ marker copy number g⁻¹ of individual host faeces. Following validation of host sensitivity and specificity, the host-associated Bacteroidales markers were detected in the Qu’Appelle Valley watershed of Saskatchewan, Canada which experiences a diversity of anthropogenic inputs. Concentrations of the ruminant marker were well-correlated with proximity to cattle operations and there was a correlation between the marker and Escherichia coli concentrations at these sites. Low concentrations of the human faecal marker were measured throughout the sampling sites, and may indicate a consistent influx of human faecal pollution into the watershed area. Persistence of each of the Bacteroidales host-associated marker was also studied in situ. The results indicated that the markers persist for shorter periods of time (99% decay in <8 days) compared with the conventional E. coli marker (99% decay in >15 days), suggesting they are effective at detecting recent faecal contamination events. The levels of Bacteroidales markers and E. coli counts did not correlate with the presence of the pathogenic bacteria, Salmonella spp. or Campylobacter spp. detected in the Qu’Appelle Valley. Collectively, the results obtained in this study demonstrated that the qPCR approach for detecting host-associated Bacteroidales spp. markers can be a useful tool in helping to determine host-specific impacts of faecal pollution into a prairie watershed.     

Evaluation of multiple sewage-associated Bacteroides PCR markers for sewage pollution tracking

The host specificity of the five published sewage-associated Bacteroides markers (i.e., HF183, BacHum, HuBac, BacH and Human-Bac) was evaluated in Southeast Queensland, Australia by testing fecal DNA samples (n = 186) from 11 animal species including human fecal samples collected via influent to a sewage treatment plant (STP). All human fecal samples (n = 50) were positive for all five markers indicating 100% sensitivity of these markers. The overall specificity of the HF183 markers to differentiate between humans and animals was 99%. The specificities of the BacHum and BacH markers were > 94%, suggesting that these markers are suitable for the detection of sewage pollution in environmental waters in Australia. The HuBac (i.e., 63%) and Human-Bac (i.e., 79% specificity) markers performed poorly in distinguishing between the sources of human and animal fecal samples. It is recommended that the specificity of the sewage-associated markers must be rigorously tested prior to its application to identify the sources of fecal pollution in environmental waters.     

Evaluating potential applications of faecal sterols in distinguishing sources of faecal contamination from mixed faecal samples

Faecal samples from humans, herbivores, carnivores and birds as well as samples from septic tanks and effluents from a sewage treatment plant (STP) were extracted and analyzed by gas chromatography-mass spectrometry for faecal sterols including coprostanol, epicoprostanol, cholestanol, cholesterol, stigmasterol, campesterol, 24-ethylcoprostanol and beta-sitosterol. Coprostanol was observed in the highest concentrations from the human derived samples, but it was also present in substantial quantities in a range of herbivores. There was no unique marker of human faecal contamination. Multivariate analyses revealed that the faecal sterol profiles were significantly different between the four groups of animals (Wilks' lambda=0.007, P<0.002), and coprostanol and 24-ethylcoprostanol were the major discriminant factors. However, when faecal samples were mixed, the confounding of faecal sterol levels prevented accurate identification of contributing species. Conversely, faecal sterol ratios were highly efficient at identifying which mixtures contained human contribution, but could not appropriately determine percentage contributions of sources.     

Isolation and characterization of Bacteroides host strain HB-73 used to detect sewage specific phages in Hawaii

Previous studies have shown that Escherichia coli and enterococci are unreliable indicators of fecal contamination in Hawaii because of their ability to multiply in environmental soils. In this study, the method of detecting Bacteroides phages as specific markers of sewage contamination in Hawaii’s recreational waters was evaluated because these sewage specific phages cannot multiply under environmental conditions. Bacteroides hosts (GB-124, GA-17), were recovered from sewage samples in Europe and were reported to be effective in detecting phages from sewage samples obtained in certain geographical areas. However, GB-124 and GA-17 hosts were ineffective in detecting phages from sewage samples obtained in Hawaii. Bacteroides host HB-73 was isolated from a sewage sample in Hawaii, confirmed as a Bacteroides sp. and shown to recover phages from multiple sources of sewage produced in Hawaii at high concentrations (5.2-7.3 × 10⁵ PFU/100 mL). These Bacteroides phages were considered as potential markers of sewage because they also survived for three days in fresh stream water and two days in marine water. Water samples from Hawaii’s coastal swimming beaches and harbors, which were known to be contaminated with discharges from streams, were shown to contain moderate (20-187 CFU/100 mL) to elevated (173-816 CFU/100 mL) concentrations of enterococci. These same samples contained undetectable levels (<10 PFU/100 mL) of F+ coliphage and Bacteroides phages and provided evidence to suggest that these enterococci may not necessarily be associated with the presence of raw sewage. These results support previous conclusions that discharges from streams are the major sources of enterococci in coastal waters of Hawaii and the most likely source of these enterococci is from environmental soil rather than from sewage.     

Impact of nitrate addition on biofilm properties and activities in rising main sewers

Anaerobic sewer biofilm is a composite of many different microbial populations, including sulfate reducing bacteria (SRB), methanogens and heterotrophic bacteria. Nitrate addition to sewers in an attempt to control hydrogen sulfide concentrations affects the behaviour of these populations, which in turn impacts on wastewater characteristics. Experiments were carried out on a laboratory reactor system simulating a rising main to determine the impact of nitrate addition on the microbial activities of anaerobic sewer biofilm. Nitrate was added to the start of the rising main during sewage pump cycles at a concentration of 30 mg-N L⁻¹ for over 5 months. While it reduced sulfide levels at the outlet of the system by 66%, nitrate was not toxic or inhibitory to SRB activity and did not affect the dominant SRB populations in the biofilm. Long-term nitrate addition in fact stimulated additional SRB activity in downstream biofilm. Nitrate addition also stimulated the activity of nitrate reducing, sulfide oxidizing bacteria that appeared to be primarily responsible for the prevention of sulfide build up in the wastewater in the presence of nitrate. A short adaptation period of three to four nitrate exposure events (approximately 10 h) was required to stimulate biological sulfide oxidation, beyond which no sulfide accumulation was observed under anoxic conditions. Nitrate addition effectively controlled methane concentrations in the wastewater. The nitrate uptake rate of the biofilm increased with repeated exposure to nitrate, which in turn increased the consumption of biodegradable COD in the wastewater. These results provide a comprehensive understanding of the impact of nitrate addition on wastewater composition and sewer biofilm microbial activities, which will facilitate optimization of nitrate dosing for effective sulfide control in rising main sewers.     

Nitrate and chloride concentrations in the high plains aquifer,Texas

Groundwater nitrate and chloride concentrations were compiled for 122 wells in a rural, three‐county area of northwest Texas. The counties are located on the High Plains aquifer, a major source of groundwater in the region. Cropland/pasture is the predominant land use in the study area. The area also contains numerous cattle feedlots. Fertilizer and manure associated with those land uses are potential sources of ground‐water contamination. Although locally elevated above background levels, none of the chemical concentrations exceeded the primary drinking water standard of 44.27 mg/L for nitrate (10mg/L for NO₃ — N) or secondary standard of 250mg/L for chloride. Rank correlations between nitrate and chloride were statistically significant in two of the three counties, where the solutes may have originated from a common surface source. Denitrification and scant precipitation recharge may account for an absence of nitrate levels above the drinking water standard.     

天然水中硝酸盐氮氧同位素测试技术研究进展

水环境的硝酸盐污染是一个全球性环境问题。识别水环境中硝酸盐的来源及其转化,对更好地管理水质是十分重要的。硝酸盐的稳定氮氧同位素组成可有效识别水环境中NO 3-来源及其转化。本文总结了典型的测定硝酸盐中稳定氮氧同位素组成的方法:石墨燃烧法、AgNO3-离子交换法、细菌反硝化法、亚硝酸盐去除联合细菌反硝化法、两步化学还原法、连续选择性细菌还原法,同时对这些方法的优缺点进行了评述。最后分析了检测技术中存在的共同问题及其发展方向:δ18O精确度仍有提高的潜力;把NO 3-从水中独立提取出来进行三氧同位素的测试方法还需改进和提高;水样的采集、保存有待进一步改进。     

The nature of the carbon source rules the competition between PAO and denitrifiers in systems for simultaneous biological nitrogen and phosphorus removal

The presence of nitrate in the theoretical anaerobic reactor of a municipal WWTP aiming at simultaneous C, N and P removal usually leads to Enhanced Biological Phosphorus Removal (EBPR) failure due to the competition between PAO and denitrifiers for organic substrate. This problem was studied in a continuous anaerobic-anoxic-aerobic (A²/O) pilot plant (146 L) operating with good removal performance and a PAO-enriched sludge (72%). Nitrate presence in the initially anaerobic reactor was studied by switching the operation of the plant to an anoxic-aerobic configuration. When the influent COD composition was a mixture of different carbon sources (acetic acid, propionic acid and sucrose) the system was surprisingly able to maintain EBPR, even with internal recycle ratios up to ten times the influent flow rate and COD limiting conditions. However, the utilisation of sucrose as sole carbon source resulted in a fast EBPR failure. Batch tests with different nitrate concentrations (0-40 mg L⁻¹) were performed in order to gain insight into the competition for the carbon source in terms of P-release or denitrification rates and P-release/C-uptake ratio. Surprisingly, no inhibitory or detrimental effect on EBPR performance due to nitrate was observed. A model based on ASM2d but considering two step nitrification and denitrification was developed and experimentally validated. Simulation studies showed that anaerobic VFA availability is critical to maintain EBPR activity.     

Electrochemical determination of pollutants in sea water: Toxic substances and nutrients

Coastal marine waters have a wide variety of important human uses and therefore must be protected against contamination. The main sources of their pollution (by toxic substances and nutrients) are the discharge of municipal sewage and runoff from agricultural areas. Excessive nutrient enrichment in the form of inorganic nitrogen (NH₄⁺, NO₂^?, NO₃^?) may be directly estimated by the use of ion selective electrodes (ISE), whereas the inorganic phosphorus can only be estimated indirectly. The total content of N, P and heavy metals may be estimated after decomposition of organic substances, using the ISE method in the case of P and N determination and the differential pulse anodic stripping voltammetry (DPASV) method for determining the heavy metals.     

Development of microbial and chemical MST tools to identify the origin of the faecal pollution in bathing and shellfish harvesting waters in France

The microbiological quality of coastal or river waters can be affected by faecal pollution from human or animal sources. An efficient MST (Microbial Source Tracking) toolbox consisting of several host-specific markers would therefore be valuable for identifying the origin of the faecal pollution in the environment and thus for effective resource management and remediation. In this multidisciplinary study, after having tested some MST markers on faecal samples, we compared a selection of 17 parameters corresponding to chemical (steroid ratios, caffeine, and synthetic compounds), bacterial (host-specific Bacteroidales, Lactobacillus amylovorus and Bifidobacterium adolescentis) and viral (genotypes I-IV of F-specific bacteriophages, FRNAPH) markers on environmental water samples (n = 33; wastewater, runoff and river waters) with variable Escherichia coli concentrations. Eleven microbial and chemical parameters were finally chosen for our MST toolbox, based on their specificity for particular pollution sources represented by our samples and their detection in river waters impacted by human or animal pollution; these were: the human-specific chemical compounds caffeine, TCEP (tri(2-chloroethyl)phosphate) and benzophenone; the ratios of sitostanol/coprostanol and coprostanol/(coprostanol+24-ethylcopstanol); real-time PCR (Polymerase Chain Reaction) human-specific (HF183 and B. adolescentis), pig-specific (Pig-2-Bac and L. amylovorus) and ruminant-specific (Rum-2-Bac) markers; and human FRNAPH genogroup II.