Quantitative source apportionment of faecal indicator bacteria from anthropogenic and zoogenic sources of faecal contamination.

Recreational bathing waters are complex systems with diverse inputs from multiple anthropogenic and zoogenic sources of faecal contamination. Faecal contamination is a substantial threat to water quality and public health. Here we present a comprehensive strategy to estimate the contribution of faecal indicator bacteria (FIB) from different biological sources on two at-risk beaches in Dublin, Ireland. The daily FIB loading rate was determined for three sources of contamination: a sewage-impacted urban stream, dog and wild bird fouling. This comparative analysis determined that the stream contributed the highest daily levels of FIB, followed by dog fouling. Dog fouling may be a significant source of FIB, contributing approximately 20 % of E. coli under certain conditions, whereas wild bird fouling contributed a negligible proportion of FIB (<3 %). This study demonstrates that source-specific quantitative microbial source apportionment (QMSA) strategies are vital to identify primary public health risks and target interventions to mitigate faecal contamination.

Inclusion of hydrodynamic properties of bathing waters is critical in selecting faecal indicators to assess public health impacts of faecal contamination.

The EU Bathing Water Directive (BWD) requires member states to assess bathing water quality according to the levels of faecal indicator bacteria (FIB) in designated bathing areas. However, this criterion has two significant limitations given that the BWD does not; (i) account for differences in hydrodynamic properties of bathing waters and, (ii) assumes that all faecal pathogens decay equally in aquatic environments. This study simulated sewage discharge events in three hypothetical aquatic environments characterised by different advection and dispersion parameters in the solute transport equation. Temporal changes in the downstream concentration of six faecal indicators were determined in simulations that utilised measured decay rates of each faecal indicator from a programme of controlled microcosm experiments in fresh and seawater environments. The results showed that the decay rates of faecal indicators are not a critical parameter in advection dominant water bodies, such as in fast-flowing rivers. Therefore, faecal indicator selection is less important in such systems and for these, FIB remains the most cost-effective faecal indicator to monitor the public health impacts of faecal contamination. In contrast, consideration of faecal indicator decay is important when assessing dispersion and advection/dispersion dominant systems, which would pertain to transitional (estuarine) and coastal waterbodies. Results suggest that the inclusion of viral indicators, such as crAssphage and PMMoV, could improve the reliability of water quality modelling and minimise the risk of waterborne illnesses from faecal contamination.

Impacts of a bacterial algicide on metabolic pathways in Chlorella vulgaris.

Chlorella is a dominant species during harmful algal blooms (HABs) worldwide, which bring about great environmental problems and are also a serious threat to drinking water safety. Application of bacterial algicides is a promising way to control HABs. However, the identified bacterial algicides against Chlorella and the understanding of their effects on algal metabolism are very limited. Here, we isolated a novel bacterium Microbacterium paraoxydans strain M1 that has significant algicidal activities against Chlorella vulgaris (algicidal rate 64.38 %, at 120 h). Atrazine-desethyl (AD) was then identified from strain M1 as an effective bacterial algicide, with inhibition or algae-lysing concentration values (EC50) of 1.64 µg/mL and 1.38 µg/mL, at 72 h and 120 h, respectively. LAD (2 µg/mL AD) or HAD (20 µg/mL AD) causes morphology alteration and ultrastructure damage, chlorophyll a reduction, gene expression regulation (for example, psbA, 0.05 fold at 24 h, 2.97 fold at 72 h, and 0.23 fold of the control in HAD), oxidative stress, lipid oxidation (MDA, 2.09 and 3.08 fold of the control in LAD and HAD, respectively, at 120 h) and DNA damage (average percentage of tail DNA 6.23 % at 120 h in HAD, slight damage: 5∼20 %) in the algal cells. The impacts of AD on algal metabolites and metabolic pathways, as well as the algal response to the adverse effects were investigated. The results revealed that amino acids, amines, glycosides and urea decreased significantly compared to the control after 24 h exposure to AD (p < 0.05). The main up-regulated metabolic pathways implied metabonomic resistance and defense against osmotic pressure, oxidative stress, photosynthesis inhibition or partial cellular structure damage, such as phenylalanine metabolism, arginine biosynthesis. The down-regulated glycine, serine and threonine metabolism is a major lead in the algicidal mechanism according to the value of pathway impact. The down-regulated glycine, and serine are responsible for the downregulation of glyoxylate and dicarboxylate metabolism, aminoacyl-tRNA biosynthesis, glutathione metabolism, and sulfur metabolism, which strengthen the algae-lysing effect. It is the first time to highlight the pivotal role of glycine, serine and threonine metabolism in algicidal activities, which provided a new perspective for understanding the mechanism of bacterial algicides exerting on algal cells at the metabolic level.

Spatial variation in bacterial biomass, community composition and driving factors across a eutrophic river.

Eutrophication is a global problem, and bacterial diversity and community composition are usually affected by eutrophication. However, limited information on the ecological significance of bacterial community during algae blooms of rivers has been given, more studies should be focused on the bacterial diversity and distribution characteristics in eutrophic rivers. In this study, we explored the spatial variations of bacterial biomass, community structure, and their relationship with environmental factors in the eutrophic Xiangxi River. The content of Chlorophyll (Chl) was about 16 mg/L in the midstream (S2, S3), which was in the range of light eutrophication. Significant spatial variation of bacterial community structure was found at different sites and depths (p < 0.05), and the driving environmental factor was found to be nitrogen, mainly detected as total nitrogen (TN), Kjeldahl nitrogen (KN), and ammonia nitrogen (NH4+) (p < 0.05). The midstream sites had some significantly different bacteria, including algicidal bacteria and dominant lineages during algal blooms. This result was consistent with the functional prediction, where significant higher abundance of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways was associated with algicidal substances in the midstream. At different water depths, some populations adapted to the surface layer, such as the class Flavobacteriia, and others preferred to inhabit in the bottom layer, such as Betaproteobacteria and Acidobacteria. The bacterial biomass was higher in the bottom layer than that in the surface and middle layer, and temperature and pH were found to be the major driving factors. The bacterial diversity increased with the increasing of depths in most sampling sites according to operational taxonomic units (OTUs), Chao1 and ACE indexes, and PO43- was demonstrated to be the most significant factor. In summary, this study offered the evidence for microbial distribution characteristics across different sites and depths in summer, and its relationship with environmental variables in a eutrophic river.

Effect of Maize Straw Biochar on Bacterial Communities in Agricultural Soil.

Biochar has become a popular soil amendment. However, its effect on soil microbial community is still unclear. In the present study, maize straw biochar was pyrolysed at 300°C, 450°C and 600°C, respectively, and then was added to agricultural soil at the ratio of 0.5%, 1% and 2%. Bacterial dynamics was analyzed in the pot experiments using denaturing gradient gel electrophoresis. The results indicated that the pyrolysis temperature has great impact on the elemental composition, pH and porous structures of biochar. Moreover, pyrolysis temperature was primary factor to drive the variation of bacterial community structure in biochar amended soil. In addition, the results suggested that biochar amendments on agricultural soil would decrease the bacterial diversity, and selectively promote growth of functional bacteria to become the dominant community, which could increase the bacterial community organization and improve the stability of bacteria to counteract effects of perturbation.

Effects of soil properties, heavy metals, and PBDEs on microbial community of e-waste contaminated soil.

Heavy metals and polybrominated diphenyl ethers (PBDEs) are ubiquitous pollutants at electronic waste (e-waste) contaminated sites, their individual impacts on soil microbial community has attracted wide attention, however, limited research is available on the combined effects of heavy metals and PBDEs on microbial community of e-waste contaminated. Therefore, combined effects of heavy metals and PBDEs on the microbial community in the e-waste contaminated soil were investigated in this study. Samples were collected from Ziya e-waste recycling area in Tianjin, northern China, and the soil microbial communities were then analyzed by the high-throughput MiSeq 16S rRNA sequencing to assess the effects of soil properties, heavy metals, and PBDEs on the soil microbial community. Candidatus Nitrososphaera, Steroidobacter and Kaistobacter were the dominant microbial species in the soils. Similar microbial metabolic functions, including amino acid metabolism, carbohydrate metabolism and membrane transport, were found in all soil samples. Redundancy analysis and variation partition analysis revealed that the microbial community was mainly influenced by PBDEs (including BDE 183, BDE 99, BDE 100 and BDE 154) in horizontal soil samples. However, TN, biomass, BDE 100, BDE 99 and BDE 66 were the major drivers shaping the microbial community in vertical soil samples.

Structural and Functional Changes of Groundwater Bacterial Community During Temperature and pH Disturbances.

In this study, we report the characteristics of a microbial community in sampled groundwater and elucidate the effects of temperature and pH disturbances on bacterial structure and nitrogen-cycling functions. The predominant phyla of candidate OD1, candidate OP3, and Proteobacteria represented more than half of the total bacteria, which clearly manifested as a "low nucleic acid content (LNA) bacteria majority" type via flow cytometric fingerprint. The results showed that LNA bacteria were more tolerant to rapid changes in temperature and pH, compared to high nucleic acid content (HNA) bacteria. A continuous temperature increase test demonstrated that the LNA bacterial group was less competitive than the HNA bacterial group in terms of maintaining their cell intactness and growth potential. In contrast, the percentage of intact LNA bacteria was maintained at nearly 70% with pH decrease, despite a 50% decrease in total intact cells. Next-generation sequencing results revealed strong resistance and growth potential of phylum Proteobacteria when the temperature increased or the pH decreased in groundwater, especially for subclasses α-, ß-, and γ-Proteobacteria. In addition, relative abundance of nitrogen-related functional genes by qPCR showed no difference in nitrifiers or denitrifiers within 0.45 µm-captured and 0.45 µm-filterable bacteria due to phylogenetic diversity. One exception was the monophyletic anammox bacteria that belong to the phylum Planctomycetes, which were mostly captured on a 0.45-µm filter. Furthermore, we showed that both temperature increase and pH decrease could enhance the denitrification potential, whereas the nitrification and anammox potentials were weakened.

Biogeography and Diversity of Freshwater Bacteria on a River Catchment Scale.

To illustrate how freshwater bacterial community changes with geographic gradient, we investigated the spatial changes of bacterial abundance and community structures from over 200 samples on a catchment scale in the Songhua River using heterotrophic plate counts, flow cytometry, denaturing gradient gel electrophoresis, and pyrosequencing analysis. The results showed that the mainstream had higher cultivable bacteria and total bacterial concentration than tributaries in the Songhua River catchment. Response model analysis demonstrated that the bacterial community exhibits a biogeographical signature even in an interconnected river network system, and the total bacterial concentration and biodiversity were significantly correlated to latitude (p < 0.001) and longitude (p < 0.001). Multivariate redundancy analysis indicated that temperature was the most important factor driving bacterial community structure in the Songhua River, which accounts for 35.30% variance of communities, then dissolved oxygen (17.60%), latitude (17.60%), longitude (11.80%), and pH (5.88%). High-throughput pyrosequencing revealed that at the phylum level, Proteobacteria was numerically dominant (89.6%) in river catchment, followed by Bacteroidetes (8.1%) and Cyanobacteria (1.2%). The overall results revealed that the bacterial community was driven by geographical distance regardless of the continuum of the river on a catchment scale.

Impact of planktonic low nucleic acid-content bacteria to bacterial community structure and associated ecological functions in a shallow lake.

In this study, 0.45 µm filtration, flow cytometric fingerprint, 16S rRNA amplicon sequencing, and bioinformation tools were adopted to analyze the structural diversity and potential functions of planktonic low nucleic acid (LNA)- content bacteria in a shallow lake. Three bacterial groups, namely, "LNA," "high nucleic acid (HNA)-Small," and "HNA-Large," were classified through flow cytometric fingerprint, among which the "HNA-Small" group was possibly in the proliferation stage of the "LNA" group. Total nitrogen and phosphate were the key factors that influence the growth of LNA bacteria. Results of 16S rRNA amplicon sequencing showed that LNA bacteria were phylogenetically less diverse than HNA bacteria, and Actinobacteria and Proteobacteria (especially Gamma-Proteobacteria) were the dominant phyla in LNA bacterial operational taxonomic units (OTUs). Accordingly, hgcI_clade and Pseudomonas were the most abundant bacterial genera in LNA bacterial OTUs. The fraction of low-abundance LNA bacteria was sensitive to several environmental factors, indicating that environmental factors only determined the fraction distribution of low-abundance bacteria. The prediction of metabolic and ecological functions showed that LNA and HNA bacteria had distinct metabolic and ecological functions, which were mainly attributed to the dominant and exclusive bacterial groups.

Resistance and resilience of representative low nucleic acid-content bacteria to free chlorine exposure.

Two representative low nucleic acid-content (LNA) bacterial strains, Polynucleobacter sp. CB and Sphingopyxis sp. 15Y-HN, and two commonly used microbial indicators of drinking water disinfection efficiency, Escherichia coli and Pseudomonas aeruginosa, were used to investigate the effects of chlorine disinfection. LNA bacteria were found to be more tolerant than microbial indicator strains at the same free chlorine concentrations in batch experiments. Three-stage chlorination experiments were carried out for two strains (15Y-HN and E. coli K12) to compare their responses to long-term chlorine exposure. Results from the first stage (increasing chlorine exposure from 0.0 to 0.6 mg/L and 1.2 mg/L for strain K12 and strain 15Y-HN, respectively) showed the rate constants of 99% cell damage were 10-fold lower for strain 15Y-HN than for strain K12. A second regrowth stage at low free chlorine concentrations (<0.3 mg/L for 140 h) facilitated the regrowth of chlorine-resistant populations of strains 15Y-HN and K12 in the presence of assimilable organic carbon (AOC). In the third stage, during which bacteria were exposed to increasing chlorine from 0.0 to 0.5-0.6 mg/L, strain 15Y-HN was maintained at 80% and 105 cells/mL of intact cells whereas strain K12 was completely damaged. The overall results demonstrated that representative LNA bacteria exhibit strong resistance and resilience to chlorine under low AOC conditions, which should be taken into consideration in disinfection processes.

Biodegradation of decabromodiphenyl ether (BDE 209) by a newly isolated bacterium from an e-waste recycling area.

Polybrominated diphenyl ethers (PBDEs) have become widespread environmental pollutants all over the world. A newly isolated bacterium from an e-waste recycling area, Stenotrophomonas sp. strain WZN-1, can degrade decabromodiphenyl ether (BDE 209) effectively under aerobic conditions. Orthogonal test results showed that the optimum conditions for BDE 209 biodegradation were pH 5, 25 °C, 0.5% salinity, 150 mL minimal salt medium volume. Under the optimized condition, strain WZN-1 could degrade 55.15% of 65 µg/L BDE 209 under aerobic condition within 30 day incubation. Moreover, BDE 209 degradation kinetics was fitted to a first-order kinetics model. The biodegradation mechanism of BDE 209 by strain WZN-1 were supposed to be three possible metabolic pathways: debromination, hydroxylation, and ring opening processes. Four BDE 209 degradation genes, including one hydrolase, one dioxygenase and two dehalogenases, were identified based on the complete genome sequencing of strain WZN-1. The real-time qPCR demonstrated that the expression level of four identified genes were significantly induced by BDE 209, and they played an important role in the degradation process. This study is the first to demonstrate that the newly isolated Stenotrophomonas strain has an efficient BDE 209 degradation ability and would provide new insights for the microbial degradation of PBDEs.

Ecological and health risks assessment and spatial distribution of residual heavy metals in the soil of an e-waste circular economy park in Tianjin, China.

Ziya Circular Economy Park is the biggest e-waste recycle park in North China before 2011, its function was then transformed in response to regulations and rules. In this paper, investigation was conducted to research the residual concentrations of 14 analytes (12 heavy metals and 2 non-metals) in the surface soil of Ziya Circular Economy Park and surrounding area. Both ecological and health assessments were evaluated using GI (geo-accumulation index) and NPI (Nemerow pollution index), and associated health risk was assessed by using USEPA model. According to the ecological risk assessment, Cu, Sb, Cd, Zn and Co were seriously enriched in the soil of the studied area. The health risk assessment proposed by USEPA indicated no significant health risks to the population. Soil properties, such as pH and organic matter, were found to correlate with the enrichment of heavy metals. Arsenic concentrations in the soil were found positively correlated to dead bacteria concentrations. Spatial distribution of heavy metals revealed that Ziya Circular Economy Park was the dominant pollution source in the studied area. Findings in this study suggest that enough attention should be payed to the heavy metal pollution in Ziya Circular Economy Park.

Spatial-Temporal Changes of Bacterioplankton Community along an Exhorheic River.

To date, few aquatic microbial ecology studies have discussed the variability of the microbial community in exorheic river ecosystems on both the spatial and seasonal scales. In this study, we examined the spatio-temporal variation of bacterioplankton community composition in an anthropogenically influenced exorheic river, the Haihe River in Tianjin, China, using pyrosequencing analysis of 16S rRNA genes. It was verified by one-way ANOVA that the spatial variability of the bacterioplankton community composition over the whole river was stronger than the seasonal variation. Salinity was a major factor leading to spatial differentiation of the microbial community structure into riverine and estuarial parts. A high temperature influence on the seasonal bacterial community variation was only apparent within certain kinds of environments (e.g., the riverine part). Bacterial community richness and diversity both exhibited significant spatial changes, and their seasonal variations were completely different in the two environments studied here. Furthermore, riverine bacterial community assemblages were subdivided into urban and rural groups due to changes in the nutritional state of the river. In addition, the nutrient-loving group including Limnohabitans, Hydrogenophaga, and Polynucleobacter were abundant in the urbanized Haihe River, indicating the environmental factors in these anthropogenic waterbodies heavily influence the core freshwater community composition.

Modelling the fate and transport of faecal bacteria in estuarine and coastal waters.

This paper details a numerical model developed to predict the fate and transport of faecal bacteria in receiving surface waters. The model was first validated by comparing model predicted faecal bacteria concentrations with available field measurements. The model simulations agreed well with the observation data. After calibration, the model was applied to investigate the effects of different parameters, including: tidal processes, river discharges from the upstream boundaries and bacteria inputs from the upstream boundaries, wastewater treatment works (WwTWs), rivers and combined sewer overflows (CSO), on the concentrations of faecal bacteria in the Ribble Estuary. The results revealed that the tide and upstream boundary bacteria inputs were the primary factors controlling the distribution of faecal bacteria. The bacteria inputs from the WwTWs in the model domain were generally found not to have a significant impact on distribution of faecal bacteria in the estuary.

Distribution and diversity of fungi in freshwater sediments on a river catchment scale.

Fungal communities perform essential functions in biogeochemical cycles. However, knowledge of fungal community structural changes in river ecosystems is still very limited. In the present study, we combined culture-dependent and culture-independent methods to investigate fungal distribution and diversity in sediment on a regional scale in the Songhua River catchment, located in North-East Asia. A total of 147 samples over the whole river catchment were analyzed. The results showed that compared to the mainstream, the tributaries have a higher fungal community organization and culturable fungal concentration, but possess lower community dynamics as assessed by denaturing gradient gel electrophoresis (DGGE). Furthermore, phylogenetic analysis of DGGE bands showed that Ascomycota and Basidiomycota were the predominant community in the Songhua River catchment. Redundancy analysis revealed that longitude was the primary factor determining the variation of fungal community structure, and fungal biomass was mainly related to the total nutrient content. Our findings provide new insights into the characteristics of fungal community distribution in a temperate zone river at a regional scale, and demonstrate that fungal dispersal is restricted by geographical barriers in a whole river catchment.

Rapid quantification of bacteria and viruses in influent, settled water, activated sludge and effluent from a wastewater treatment plant using flow cytometry.

As microbiological parameters are important in monitoring the correct operation of wastewater treatment plants and controlling the microbiological quality of wastewater, the abundances of total bacteria (including intact and damaged bacterial cells) and total viruses in wastewater were investigated using a combination of ultrasonication and flow cytometry. The comparisons between flow cytometry (FCM) and other cultivation-independent methods (adenosine tri-phosphate (ATP) analysis for bacteria enumeration and epifluorescence microscopy (EFM) for virus enumeration) gave very similar patterns of microbial abundance changes, suggesting that FCM is suitable for targeting and obtaining reliable counts for bacteria and viruses in wastewater samples. The main experimental results obtained were: (1) effective removal of total bacteria in wastewater, with a decrease from an average concentration of 1.74 × 10(8)counts ml(-1) in raw wastewater to 3.91 × 10(6)counts ml(-1) in the effluent, (2) compared to influent raw wastewater, the average concentration of total viruses in the treated effluent (3.94 × 10(8)counts ml(-1)) exhibited no obvious changes, (3) the applied FCM approach is a rapid, easy, and convenient tool for understanding the microbial dynamics and monitoring microbiological quality in wastewater treatment processes.

Modelling importance of sediment effects on fate and transport of enterococci in the Severn Estuary, UK.

The paper detailed a water quality modelling study of a hyper-tidal estuary, undertaken to assess the impact of various bacteria input loads on the receiving waters in a coastal basin in the UK, by using the model developed in previous study of the same authors enterococci, used as the indicators for bathing water quality under the new European Union (EU) Bathing Water Directive, were numerically modelled using a hydro-environmental model. In particular, the numerical model used in this study includes the effects of sediment on bacteria transport processes in surface water. Finally, the importance of sediment bacteria inputs on the bathing water quality was also investigated under different weather and tidal condition. During spring tide, the bacteria input from the bed sediments are dominant for both wet and dry weather conditions. During neap tides and during dry weather conditions the inputs of bacteria from the bed sediment were still dominant, but during wet weather conditions the inputs from river were dominant. Under different tidal flow conditions some parameters had a more significant role than others. During high flow conditions the sediment re-suspensions processes were dominant, therefore the bed bacteria concentrations played a dominant role on the overall bacteria concentration levels in the water column. In contrast, during low flow conditions sediment deposition prevails and bacteria are removed from the water column. The partition coefficient was found to be more important than the bed bacteria concentrations, during low flow conditions.

Numerical modelling of sediment-bacteria interaction processes in surface waters.

Faecal bacteria exist in both free-living and attached forms in surface waters. The deposition of sediments can take faecal bacteria out of the water column and to the bed. The sediments can subsequently be re-suspended into the water column, which can then lead to the re-suspension of the faecal bacteria of the attached form back into the water column, where it may desorb from the sediments. Therefore, the fate and transport of faecal bacteria is highly related to the governing sediment transport processes, particularly where these processes are significant. However, little attempt has been made to model such processes in terms of predicting the impact of the sediment fluxes on faecal bacteria levels. Details are given of the refinement of a numerical model of faecal bacteria transport, where the sediment transport processes are significant. This model is based on the model DIVAST (Depth Integrated Velocities And Solute Transport). Analytical solutions for steady and uniform flow conditions were derived and used to test the sediment-bacteria interaction model. After testing the sediment-bacteria interaction model favourably against known results, the model was then set up for idealized case studies to investigate the effects of sediment on bacteria concentrations in the water column. Finally the model was applied to a simplified artificial flooding study to investigate the impact of suspended sediment fluxes on the corresponding bacteria transport processes. The model predictions have proved to be encouraging, with the results being compared to field measurements.