Jiangyong Hu

Acyl-homoserine lactone acylase from Ralstonia strain XJ12B represents a novel and potent class of quorum-quenching enzymes

N ‐acylhomoserine lactones (AHLs) are used as signal molecules by many quorum‐sensing Proteobacteria. Diverse plant and animal pathogens use AHLs to regulate infection and virulence functions. These signals are subject to biological inactivation by AHL‐lactonases and AHL‐acylases. Previously, little was known about the molecular details underlying the latter mechanism. An AHL signal‐inactivating bacterium, identified as a Ralstonia sp., was isolated from a mixed‐species biofilm. The signal inactivation encoding gene from this organism, which we call aiiD , was cloned and successfully expressed in Escherichia coli and inactivated three AHLs tested. The predicted 794‐amino‐acid polypeptide was most similar to the aculeacin A acylase (AAC) from Actinoplanes utahensis and also shared significant similarities with cephalosporin acylases and other N‐terminal (Ntn) hydrolases. However, the most similar homologues of AiiD are deduced proteins of undemonstrated function from available Ralstonia , Deinococcus and Pseudomonas genomes. LC‐MS analyses demonstrated that AiiD hydrolyses the AHL amide, releasing homoserine lactone and the corresponding fatty acid. Expression of AiiD in Pseudomonas aeruginosa PAO1 quenched quorum sensing by this bacterium, decreasing its ability to swarm, produce elastase and pyocyanin and to paralyse nematodes. Thus, AHL‐acylases have fundamental implications and hold biotechnological promise in quenching quorum sensing.

Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) in sewage treatment plants.

In this study, the concentrations of PFOS and PFOA in the biological units of various full-scale municipal sewage treatment plants were measured. Samples of influent, primary effluent, aeration tank effluent, final effluent and grab samples of primary, activated, secondary and anaerobically digested sludge were collected by 5 sampling events over one year. The two sewage treatment plants (STPs) selected for this study include plant A receiving 95% domestic wastewater and plant B receiving 60% industrial wastewater and 40% domestic wastewater. PFOS and PFOA were observed at higher concentration in aqueous and sludge samples in plant B than that of plant A. Mass flow of PFOS increased significantly (mean 94.6%) in conventional activated sludge process (CAS) of plant B, while it remained consistent after the secondary treatment in plant A. Mass flow of PFOA increased 41.6% (mean) in CAS of plants A and B and 76.6% in membrane biological reactor (MBR), while it remained unchanged after the treatment of liquid treatment module (LTM). Our results suggest that mass flow of these two compounds remains consistent after treatment of activated sludge process operating at short sludge retention time (SRT). Seasonal variations of PFOS in concentrations of raw sewage were found in plant A, while PFOA did not have significant seasonal variation in both plants A and B.

Quorum quenching enzyme activity is widely conserved in the sera of mammalian species

Acyl‐homoserine lactone (AHL) quorum sensing signals play a key role in synchronizing virulence gene expression in Pseudomonas aeruginosa, which could cause fatal bloodstream infections. We showed that AHL inactivation activity, albeit with variable efficiency, was conserved in the serum samples of all the 6 tested mammalian animals. High‐performance liquid chromatography and mass spectrometry analyses revealed that mammalian sera had a lactonase‐like enzyme(s), which hydrolyzed the lactone ring of AHL to produce acyl homoserine, with enzyme properties reminiscent of paraoxonases (PONs). We further showed that the animal cell lines expressing three mouse PON genes, respectively, displayed strong AHL degradation activities.

Insight into metabolic and cometabolic activities of autotrophic and heterotrophic microorganisms in the biodegradation of emerging trace organic contaminants

Many efforts have been made to understand the biodegradation of emerging trace organic contaminants (EOCs) in the natural and engineered systems. This review summarizes the current knowledge on the biodegradation of EOCs while having in-depth discussion on metabolism and cometabolism of EOCs. Biodegradation of EOCs is mainly attributed to cometabolic activities of both heterotrophic and autotrophic microorganisms. Metabolism of EOCs can only be observed by heterotrophic microbes. Autotrophic ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaeal (AOA) cometabolize a variety of EOCs via the non-specific enzymes, such as ammonia monooxygenase (AMO). Higher biodegradation of EOCs is often noted under nitrification at high ammonia loading rate. The presence of a growth substrate promotes cometabolic biodegradation of EOCs. Potential strategies for enhancing the biodegradation of EOCs were also proposed in this review.

Investigation of assimilable organic carbon (AOC) and bacterial regrowth in drinking water distribution system

This paper investigated the variation of assimilable organic carbon (AOC) concentrations in water from several typical water treatment plants and distribution systems in a northern city of China. It is concluded from this study that: (1) The AOC in most of the product water of the studied water treatment plants and the water from the associated distribution systems could not meet the biostability criteria of 50-100 microg/L. (2) Only 4% of the measured AOC concentrations were less than 100 microg/L. However, about half of the measured AOC values were less than 200 microg/L. (3) Better source water quality resulted in lower AOC concentrations. (4) The variation of AOC concentrations in distribution systems was affected by chlorine oxidation and bacterial activity: the former resulted in an increase of AOC value while the latter led to a reduction in AOC. (5) The variation of AOC concentration followed different patterns in different distribution systems or different seasons due to their respective operational characteristics. (6) Less than 30% of AOC could be removed by a conventional treatment process, whereas 30-60% with a maximum of 50-60% could be removed by granular activated carbon (GAC). (7) The observation via scanning electron microscope (SEM) on distribution pipe tubercle samples demonstrated that the pipe inner wall was not smooth and bacteria multiplied in the crevice as well as in the interior wall of distribution pipes.

A new method for characterizing denitrifying phosphorus removal bacteria by using three different types of electron acceptors.

This study investigated the characteristics of denitrifying phosphorus removal bacteria by using three different types of electron acceptors as well as the positive role of nitrite in phosphorus removal process. Denitrifying phosphorous removal bacteria was enriched under anaerobic-anoxic (A/A) condition. To understand A/A sludge better, sludge from two other sources were also studied. These include sludges obtained from a lab-scale anaerobic-anoxic-aerobic (A/A/O) system and a local sewage treatment plant. Three types of possible electron acceptors (oxygen, nitrate and nitrite) were examined for their roles in phosphorus uptake. The results obtained indicated that oxygen, nitrate and nitrite were able to act as electron acceptors successfully. This observation suggested that in addition to the two well-accepted groups of phosphorus removal bacteria (one can only utilize oxygen to take up phosphorus, P(O), while the other can use both oxygen and nitrate, P(ON)), a new group of phosphorus removal bacteria, P(ON(n)), which could use oxygen, nitrate or nitrite to take up phosphorus was identified. The relative population of these three types of bacteria could be calculated from results obtainable from phosphorus uptake batch experiments with either oxygen or nitrate or nitrite as electron acceptor. The results obtained in this study showed that A/A sludge had similar phosphorus removal performance as the A/A/O sludge. However, it has better denitrifying phosphorus removal capability, which was demonstrated by the relative population of the three groups of bacteria. The results also suggested that nitrite was not an inhibitor to phosphorus removal process. Instead, it is an alternative electron acceptor to oxygen or nitrate.

Simultaneous determination of PPCPs, EDCs, and artificial sweeteners in environmental water samples using a single-step SPE coupled with HPLC-MS/MS and isotope dilution.

A high-throughput method for the simultaneous determination of 24 pharmaceuticals and personal care products (PPCPs), endocrine disrupting chemicals (EDCs) and artificial sweeteners (ASs) was developed. The method was based on a single-step solid phase extraction (SPE) coupled with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and isotope dilution. In this study, a single-step SPE procedure was optimized for simultaneous extraction of all target analytes. Good recoveries (≥ 70%) were observed for all target analytes when extraction was performed using Chromabond(®) HR-X (500 mg, 6 mL) cartridges under acidic condition (pH 2). HPLC-MS/MS parameters were optimized for the simultaneous analysis of 24 PPCPs, EDCs and ASs in a single injection. Quantification was performed by using 13 isotopically labeled internal standards (ILIS), which allows correcting efficiently the loss of the analytes during SPE procedure, matrix effects during HPLC-MS/MS and fluctuation in MS/MS signal intensity due to instrument. Method quantification limit (MQL) for most of the target analytes was below 10 ng/L in all water samples. The method was successfully applied for the simultaneous determination of PPCPs, EDCs and ASs in raw wastewater, surface water and groundwater samples collected in a local catchment area in Singapore. In conclusion, the developed method provided a valuable tool for investigating the occurrence, behavior, transport, and the fate of PPCPs, EDCs and ASs in the aquatic environment.

Ozone-biological activated carbon as a pretreatment process for reverse osmosis brine treatment and recovery

Ozonation was used in this study to improve biodegradability of RO brine from water reclamation facilities. An ozone dosage ranging from 3 to 10 mg O(3)/L and contact times of 10 and 20 min in batch studies were found to increase the biodegradability (BOD(5)/TOC ratio) of the RO brine by 1.8-3.5 times. At the same time, total organic carbon (TOC) removal was in the range of 5.3-24.5%. The lab-scale ozone-biological activated carbon (BAC) at an ozone dosage of 6.0mg O(3)/L with 20-min contact time was able to achieve 3 times higher TOC removal compared to using BAC alone. Further processing with Capacitive Deionization (CDI) process was able to generate a product water with better water quality than the RO feed water, i.e., with more than 80% ions removal and a lower TOC concentration. The ozone-BAC pretreatment has the potential of reducing fouling in the CDI process.

Effects of pH and temperature on the survival of coliphages MS2 and Qβ

The RNA F-specific coliphages, MS2 and Qβ, have been used as virus indicators in water and wastewater studies. It is therefore useful to have a good understanding concerning the effects of environmental factors on their survival in order to choose an appropriate candidate for assessing microbial safety in relation to water quality management. The effects of pH and temperature on the survival of these two coliphages were investigated. MS2 survived better in acidic conditions than in an alkaline environment. In contrast, Qβ had a better survival rate in alkaline conditions than in an acidic environment. The inactivation rates of both coliphages were lowest within the pH range 6–8 and the temperature range 5–35°C. The inactivation rates of both coliphages increased when the pH was decreased to below 6 or increased to above 8. The inactivation rates of both coliphages increased with increasing temperature. Qβ behaved peculiarly in extreme pH buffers, i.e. it was inactivated very rapidly initially when subjected to an extreme pH environment, although the inactivation rate subsequently decreased. In general, MS2 was a better indicator than Qβ. However, within the pH range 6–9 and at temperatures not above 25°C, either MS2 or Qβ could be used as a viral indicator.

Suitability of artificial sweeteners as indicators of raw wastewater contamination in surface water and groundwater.

There is no quantitative data on the occurrence of artificial sweeteners in the aquatic environment in Southeast Asian countries, particularly no information on their suitability as indicators of raw wastewater contamination on surface water and groundwater. This study provided the first quantitative information on the occurrence of artificial sweeteners in raw wastewater, surface water and groundwater in the urban catchment area in Singapore. Acesulfame, cyclamate, saccharin, and sucralose were ubiquitous in raw wastewater samples at concentrations in the range of ng/L-μg/L, while other sweeteners were not found or found only in a few of the raw wastewater samples. Residential and commercial effluents were demonstrated to be the two main sources of artificial sweeteners entering the municipal sewer systems. Relatively higher concentrations of the detected sweeteners were frequently found in surface waters at the sampling sites located in the residential/commercial areas. No significant difference in the concentrations of the detected sweeteners in surface water or groundwater was noted between wet and dry weather conditions (unpaired T-test, p> 0.05). Relatively higher concentrations and detection frequencies of acesulfame, cyclamate and saccharin in surface water samples were observed at the potentially impacted sampling sites, while these sweeteners were absent in most of the background surface water samples. Similarly, acesulfame, cyclamate, and saccharin were found in most groundwater samples at the monitoring well (GW6), which is located close to known leaking sewer segment; whereas these were absent in the background monitoring well, which is located in the catchment with no known wastewater sources. Taken together, the results suggest that acesulfame, cyclamate, and saccharin can be used as potential indicators of raw wastewater contamination in surface water and groundwater.