Transcriptome Profiling of Stenotrophomonas sp. Strain WZN-1 Reveals Mechanisms of 2,2',4,4'-Tetrabromodiphenyl Ether (BDE-47) Biotransformation.

The brominated flame retardant 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) is extensively used, stable, and difficult to degrade in the environment. The existence of BDE-47 could pose a certain risk to the environment and human health. However, the biotransformation mechanisms of BDE-47 by microorganisms remain unclear. In this study, aerobic degradation of BDE-47 by Stenotrophomonas sp. strain WZN-1 and transcriptome analysis were carried out. BDE-47 degradation by Stenotrophomonas sp. strain WZN-1 was mainly through the biological action of intracellular enzymes via the route of debromination and hydroxylation. The results of the transcriptome sequencing indicated the differentially expressed genes were related to transport, metabolism, and stress response. The key processes involved the microbial transmembrane transportation of BDE-47, energy anabolism, synthesis, and metabolism of functional enzymes, stress response, and other biological processes of gene regulation. In particular, bacterial chemotaxis played a potential role in biodegradation of BDE-47 by Stenotrophomonas sp. strain WZN-1. This study provides the first insights into the biotransformation of Stenotrophomonas sp. strain WZN-1 to BED-47 stress and shows potential for application in remediation of polluted environments.

Automated flow cytometry as a flexible tool for comparing disinfection characteristics of indigenous bacterial communities and pure cultures.

Bacterial inactivation efficiency of chlorine varies with organisms and environmental conditions. The comparability of different samples/studies, especially comparing indigenous bacterial communities with pure cultures, is impeded by inconsistent experimental conditions and analytical methods used in various studies. We tested a novel 96-well plate FCM experimental and automated analytical approach, where bacterial communities and pure cultures were suspended in the same natural water matrix prior to chlorination directly in the plate. We demonstrated the ability to rapidly monitor the efficiency of 32 different combinations of chlorine concentration and time (i.e. chlorine exposure) on bacterial pure cultures and indigenous aquatic communities, which enabled correct comparison of the data from different samples under the exact same experimental conditions. In this study, the 96-well plate automated FCM approach enabled large sets (896) of independent chlorination experiments to be carried out in a short time period. To our knowledge, this is the largest dataset of chlorination experiments which consumed least time (within 18 h after sampling) until now. Staining with SYBR Green I (SG) and SG combined with propidium iodide (SGPI) was used to assess cellular damage during chlorination. The results showed that with the same chlorine exposure, a higher chlorine concentration with a shorter contact time is favorable for inactivation of bacteria. Our research provides a promising framework to compare disinfection characteristics of various microorganism and can be further developed to diagnose effect of antimicrobial products.

Epilithic biofilm as a reservoir for functional virulence factors in wastewater-dominant rivers after WWTP upgrade.

Virulence factors (VFs) confer upon pathogens the ability to cause various types of damage or diseases. Wastewater treatment plants (WWTPs) are important point sources for the emission of pathogens and VFs into receiving rivers. Conventional WWTP upgrades are often implemented to improve the water quality of receiving ecosystems. However, knowledge on the pathogens, VFs, and health risks to receiving aquatic ecosystems after upgrade remains limited. In this study, we investigated detailed pathogenic information, including taxa, pathogenicity, and health risk, in two wastewater-dominant rivers after WWTP upgrade. Using 16S rRNA gene sequencing, we screened 14 potential pathogens in water and epilithic biofilm samples, though they were significantly more enriched in the biofilms. Combining 16S rRNA and metagenomic sequencing data, we identified Pseudomonas and Aeromonas as the dominant pathogenic taxa carrying functional VFs (e.g., mobility and offensive) in the epilithic biofilm. Moreover, strong pathogen-specific VF-host co-occurrence events were observed in the epilithic biofilm samples, indicating the importance of biofilms as reservoirs and vehicles for VFs. Further, we demonstrated that mobility VF is crucial for biofilm formation and pathogens in biofilm carrying offensive VF may be highly invasive. Quantification and health risk assessment suggested that the skin contact risk of P. aeruginosa carrying VFs was higher than the acceptable probability of 10-4 in both water and epilithic biofilm samples, which may threaten ecological and human health.

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.

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.

Crystallization and preliminary X-ray characterization of an NAD(P)-dependent butanol dehydrogenase A from Geobacillus thermodenitrificans NG80-2.

Geobacillus thermodenitrificans NG80-2 encodes two long-chain NAD(P)-dependent alcohol dehydrogenases, gtADH1 and gtADH2, in the terminal oxidation pathway of long-chain n-alkanes for the conversion of long-chain alkyl alcohols to their corresponding aldehydes. Both gtADH1 and gtADH2 are thermostable enzymes and oxidize long-chain alkyl alcohols up to at least C(30). In order to understand the structural basis for their role in long-chain alkane degradation, we have crystallized gtADH2. Single, colourless crystals were obtained from a recombinant preparation of ADH2 overexpressed in Escherichia coli. The crystals belong to space group C222(1), with unit-cell parameters a = 56.0, b = 99.6, c = 123.1 Å. Diffraction data were collected in-house to 1.79 Å resolution. The crystals contain one monomer in the asymmetric unit, with a V(M) value of 2.17 Å(3) Da(-1) and an estimated solvent content of 43%.

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.

Microcystis spp. and phosphorus in aquatic environments: A comprehensive review on their physiological and ecological interactions.

Cyanobacterial blooms caused by eutrophication have become a major environmental problem in aquatic ecosystems worldwide over the last few decades. Phosphorus is a limiting nutrient that affects the growth of cyanobacteria and plays a role in dynamic changes in algal density and the formation of cyanobacterial blooms. Therefore, identifying the association between phosphorus sources and Microcystis, which is the most representative and harmful cyanobacteria, is essential for building an understanding of the ecological risks of cyanobacterial blooms. However, systematic reviews summarizing the relationships between Microcystis and phosphorus in aquatic environments are rare. Thus, this study provides a comprehensive overview of the physiological and ecological interactions between phosphorus sources and Microcystis in aquatic environments from the following perspectives: (i) the effects of phosphorus source and concentration on Microcystis growth, (ii) the impacts of phosphorus on the environmental behaviors of Microcystis, (iii) mechanisms of phosphorus-related metabolism in Microcystis, and (iv) role of Microcystis in the distribution of phosphorus sources within aquatic environments. In addition, relevant unsolved issues and essential future investigations (e.g., secondary ecological risks) have been highlighted and discussed. This review provides deeper insights into the relationship between phosphorus sources and Microcystis and can serve as a reference for the evaluation, monitoring, and effective control of cyanobacterial blooms.

Monsoon affects the distribution of antibiotic resistome in Tibetan glaciers.

Antibiotic-resistance gene (ARG) is a biological pollutant and is globally distributed due to increased anthropogenic activities. ARGs in the cryosphere have received increased attention due to global warming, and ARGs in glaciers are predicted to be released into downstream ecosystems during glacier melting. In this study, ARG distribution and influential factors were investigated in 85 samples from 21 Tibetan glaciers, covering snow, ice, and cryoconite habitats. The results revealed ARGs against 29 antibiotics in Tibetan glaciers, dominated by tetracycline, bacitracin, macrolide, and fluoroquinolone resistance. ARGs in snow exhibited biogeographic patterns influenced by atmospheric circulation. Specifically, monsoon-dominated glaciers exhibited a significantly higher abundance of ARGs than the westerly-dominated glaciers, which could be associated with higher antibiotic usage in the Indian subcontinent. Of the 3241 metagenome-assembled genomes obtained, 36.8% of which were identified as ARG hosts and 33.8% were multidrug-resistant. In addition, 90 ARGs were linked to mobile genetic elements (MGEs). 90.9% and 9.1% of MGEs were identified as plasmid and phage in 45 MAGs carrying both ARGs and MGEs. Our study suggests a greater risk of ARGs being released from the monsoon-dominated glaciers, which were the glaciers that melt at high rates and thus need to be carefully monitored.

Variation of High and Low Nucleic Acid-Content Bacteria in Tibetan Ice Cores and Their Relationship to Black Carbon.

Nutrient enrichment caused by black carbon (BC) is a major ecological crisis in glacial ecosystems. The microbiological effects of BC were assessed in this study by using fluorescent fingerprinting assay based on flow cytometry (FCM) of bacterial communities with low (LNA) and high (HNA) nucleic acid-content bacteria. Here, we investigated a high-resolution temporal variation of bacterial abundance and LNA/HNA ratio in Tibetan ice cores. Our results revealed that bacterial abundance was proportional to the atmospheric BC on the glaciers. The shift of LNA functional groups to HNA functional groups in glaciers suggested BC emissions increased the proportion of highly active cells. In addition, distinct number of LNA and HNA functional groups was identified between the monsoon and non-monsoon seasons. Westerly winds with high amounts of BC accounted for high ratio of HNA functional groups during the non-monsoon season. In comparison, high moisture during the monsoon season decreased atmospheric BC loading, which increases the ratio of LNA functional groups. Correlations between BC and functional groups were very strong, showing that two functional groups may serve as early-warning indicators of microbiological effects of BC at low trophic level. Our approach provides a potential early-warning framework to study the influences of atmospheric BC on the glaciological community.

Disruption and recovery of river planktonic community during and after the COVID-19 outbreak in Wuhan, China.

During the COVID-19 outbreak in Wuhan, large amounts of anti-coronavirus chemicals, such as antiviral drugs and disinfectants were discharged into the surrounding aquatic ecosystem, causing potential ecological damage. Here, we investigated plankton in the Wuhan reaches of the Yangtze River, before, during, and after COVID-19, with the river reaches of three adjacent cities sampled for comparison. During the COVID-19, planktonic microbial density declined significantly. Correspondingly, the eukaryotic and prokaryotic community compositions and functions shifted markedly, with increasing abundance of chlorine-resistant organisms. Abundance of antibiotic resistance genes, virulence factor genes, and bacteria containing both genes increased by 2.3-, 2.7-, and 7.9-fold, respectively, compared to other periods. After COVID-19, all measured plankton community compositional and functional traits recovered in the Yangtze River.

Machine learning approach identifies water sample source based on microbial abundance.

Water quality can change along a river system due to differences in adjacent land use patterns and discharge sources. These variations can induce rapid responses of the aquatic microbial community, which may be an indicator of water quality characteristics. In the current study, we used a random forest model to predict water sample sources from three different river ecosystems along a gradient of anthropogenic disturbance (i.e., less disturbed mountainous area, wastewater discharged urban area, and pesticide and fertilizer applied agricultural area) based on environmental physicochemical indices (PCIs), microbiological indices (MBIs), and their combination. Results showed that among the PCI-based models, using conventional water quality indices as inputs provided markedly better prediction of water sample source than using pharmaceutical and personal care products (PPCPs), and much better prediction than using polycyclic aromatic hydrocarbons (PAHs) and substituted PAHs (SPAHs). Among the MBI-based models, using the abundances of the top 30 bacteria combined with pathogenic antibiotic resistant bacteria (PARB) as inputs achieved the lowest median out-of-bag error rate (9.9%) and increased median kappa coefficient (0.8694), while adding fungal inputs reduced the kappa coefficient. The model based on the top 30 bacteria still showed an advantage compared with models based on PCIs or the combination of PCIs and MBIs. With improvement in sequencing technology and increase in data availability in the future, the proposed method provides an economical, rapid, and reliable way in which to identify water sample sources based on abundance data of microbial communities.

Identification and quantification of bacterial genomes carrying antibiotic resistance genes and virulence factor genes for aquatic microbiological risk assessment.

Aquatic ecosystems have been increasingly threatened by anthropogenic activities, e.g., wastewater discharge and farm operation. Several methods are adopted to evaluate the effects of anthropogenic activities on biological risk in the environment, such as qPCR and amplicon next-generation sequencing. However, these methods fall short of providing genomic information of target species, which is vital for risk assessment from genomic aspect. Here, we developed a novel approach integrating metagenomic analysis and flow cytometry to identify and quantify potential pathogenic antibiotic resistant bacteria (PARB; carrying both antibiotic resistance genes (ARGs) and virulence factor genes (VFGs)) in the environment, which are of particular concern due to their infection ability and antibiotic resistance. Based on the abundance/density of PARB, we evaluated microbiological risk in a river impacted by both municipal drainage and agriculture runoff. We collected samples upstream (mountainous area) as the control. Results showed that 81.8% of dominant PARB (33) recovered using our approach were related to known pathogenic taxa. In addition, intragenomic ARGs-VFGs coexistence patterns in the dominant Pseudomonas genomes (20 out of 71 PARB) showed high similarity with the most closely related Pseudomonas genomes from the NCBI RefSeq database. These results reflect acceptable reliability of the approach for (potential) pathogen identification in environmental samples. According to the PARB density, microbiological risk in samples from the agricultural area was significantly higher than in samples from the urban area. We speculated that this was due to the higher antibiotic usage in agriculture as well as intragenomic ARGs-VFGs co-evolution under antibiotic selective pressure. This study provides an alternative approach for the identification and quantification of PARB in aquatic environments, which can be applied for microbiological risk assessment.

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.

Short-term organic carbon migration from polymeric materials in contact with chlorinated drinking water.

Polymeric materials are widely used in drinking water distribution systems. These materials could release organic carbon that supports bacterial growth. To date, the available migration assays for polymeric materials have not included the potential influence of chlorination on organic carbon migration behavior. Hence, we established a migration and growth potential protocol specifically for analysis of carbon migration from materials in contact with chlorinated drinking water. Four different materials were tested, including ethylene propylene dienemethylene (EPDM), poly-ethylene (PEX b and PEX c) and poly-butylene (PB). Chlorine consumption rates decreased gradually over time for EPDM, PEXc and PB. In contrast, no free chlorine was detected for PEXb at any time during the 7 migration cycles. Total organic carbon (TOC) and assimilable organic carbon (AOC) was evaluated in both chlorinated and non-chlorinated migrations. TOC concentrations for EPDM and PEXb in chlorinated migrations were significantly higher than non-chlorinated migrations. The AOC results showed pronounced differences among tested materials. AOC concentrations from chlorinated migration waters of EPDM and PB were higher compared to non-chlorinated migrations, whereas the opposite trend was observed for PEXb and PEXc. There was also a considerable difference between tested materials with regards to bacterial growth potential. The results revealed that the materials exposed to chlorine-influenced migration still exhibited a strong biofilm formation potential. The overall results suggested that the choice in material would make a considerable difference in chlorine consumption and carbon migration behavior in drinking water distribution systems.

Long-Term Effects of Residual Chlorine on Pseudomonas aeruginosa in Simulated Drinking Water Fed With Low AOC Medium.

Residual chlorine is often required to remain present in public drinking water supplies during distribution to ensure water quality. It is essential to understand how bacteria respond to long-term chlorine exposure, especially with the presence of assimilable organic carbon (AOC). This study aimed to investigate the effects of chlorination on Pseudomonas aeruginosa in low AOC medium by both conventional plating and culture-independent methods including flow cytometry (FCM) and quantitative PCR (qPCR). In a simulated chlorinated system using a bioreactor, membrane damage and DNA damage were measured by FCM fluorescence fingerprint. The results indicated membrane permeability occurred prior to DNA damage in response to chlorination. A regrowth of P. aeruginosa was observed when the free chlorine concentration was below 0.3 mg/L. The bacterial response to long-term exposure to a constant low level of free chlorine (0.3 mg/L) was subsequently studied in detail. Both FCM and qPCR data showed a substantial reduction during initial exposure (0-16 h), followed by a plateau where the cell concentration remained stable (16-76 h), until finally all bacteria were inactivated with subsequent continuous chlorine exposure (76-124 h). The results showed three-stage inactivation kinetics for P. aeruginosa at a low chlorine level with extended exposure time: an initial fast inactivation stage, a relatively stable middle stage, and a final stage with a slower rate than the initial stage. A series of antibiotic resistance tests suggested long-term exposure to low chlorine level led to the selection of antibiotic-resistant P. aeruginosa. The combined results suggest that depletion of residual chlorine in low AOC medium systems could reactivate P. aeruginosa, leading to a possible threat to drinking water safety.

Laboratory-Scale Simulation and Real-Time Tracking of a Microbial Contamination Event and Subsequent Shock-Chlorination in Drinking Water.

Rapid contamination of drinking water in distribution and storage systems can occur due to pressure drop, backflow, cross-connections, accidents, and bio-terrorism. Small volumes of a concentrated contaminant (e.g., wastewater) can contaminate large volumes of water in a very short time with potentially severe negative health impacts. The technical limitations of conventional, cultivation-based microbial detection methods neither allow for timely detection of such contaminations, nor for the real-time monitoring of subsequent emergency remediation measures (e.g., shock-chlorination). Here we applied a newly developed continuous, ultra high-frequency flow cytometry approach to track a rapid pollution event and subsequent disinfection of drinking water in an 80-min laboratory scale simulation. We quantified total (TCC) and intact (ICC) cell concentrations as well as flow cytometric fingerprints in parallel in real-time with two different staining methods. The ingress of wastewater was detectable almost immediately (i.e., after 0.6% volume change), significantly changing TCC, ICC, and the flow cytometric fingerprint. Shock chlorination was rapid and detected in real time, causing membrane damage in the vast majority of bacteria (i.e., drop of ICC from more than 380 cells µl-1 to less than 30 cells µl-1 within 4 min). Both of these effects as well as the final wash-in of fresh tap water followed calculated predictions well. Detailed and highly quantitative tracking of microbial dynamics at very short time scales and for different characteristics (e.g., concentration, membrane integrity) is feasible. This opens up multiple possibilities for targeted investigation of a myriad of bacterial short-term dynamics (e.g., disinfection, growth, detachment, operational changes) both in laboratory-scale research and full-scale system investigations in practice.

Structure and characterization of a NAD(P)H-dependent carbonyl reductase from Pseudomonas aeruginosa PAO1.

To investigate the function of the pa4079 gene from the opportunistic pathogen Pseudomonas aeruginosa PAO1, we determined its crystal structure and confirmed it to be a NAD(P)-dependent short-chain dehydrogenase/reductase. Structural similarity and activity for a broad range of substrates indicate that PA4079 functions as a carbonyl reductase. Comparison of apo- and holo-PA4079 shows that NADP stabilizes the active site specificity loop, and small molecule binding induces rotation of the Tyr183 side chain by approximately 90° out of the active site. Quantitative real-time PCR results show that pa4079 maintains high expression levels during antibiotic exposure. This work provides a starting point for understanding substrate recognition and selectivity by PA4079, as well as its possible reduction of antimicrobial drugs. DATABASE: Structural data are available in the Protein Data Bank (PDB) under the following accession numbers: apo PA4079 (condition I), 5WQM; apo PA4079 (condition II), 5WQN; PA4079 + NADP (condition I), 5WQO; PA4079 + NADP (condition II), 5WQP.

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.