Effect of DNA sizes and reactive oxygen species on degradation of sulphonamide resistance sul1 genes by combined UV/free chlorine processes.

Nowadays, antibiotic resistance genes (ARGs) have been characterized as an emerging environmental contaminant, as the spread of ARGs may increase the difficulty of bacterial infection treatments. This study evaluates the combination of ultraviolet (UV) irradiation and chlorination, the two most commonly applied disinfection methods, on the degradation of sulphonamide resistance sul1 genes. The results revealed that although both of individual UV and chlorination processes were relatively less effective, two of the four combined processes, namely UV followed by chlorination (UV-Cl2) and simultaneous combination of UV and chlorination (UV/Cl2), delivered a better removal rate (up to 1.5 logs) with an observation of synergetic effects up to 0.609 log. The mechanisms analysis found that the difference of DNA size affected sul1 genes degradation by UV and chlorination; targeted genes on larger DNA fragments could be more effectively degraded by UV (1.09 logs for large fragments and 0.12 log for small fragments when UV dose reached 432 mJ/cm2), while to degrade ARGs on smaller DNA fragments required less free chlorine dosage (10 mg/L for small fragments and 40 mg/L for large fragments). The sequential combination of UV and chlorination (UV-Cl2) used the corresponding reactivity of both processes, which could be the reason for the synergetic effect. For UV/Cl2 process, the formation of reactive oxygen species (ROS) contributed to the synergetic effect. Scavenger analysis showed that the contribution of ROS to the sul1 gene reduction was 0.004 to 0.273 log (up to 45.5 % of the total synergy values), and among the two major reactive species in UV/Cl2 system, HO was the more important radical, while the contribution of Cl was negligible. Besides, UV/Cl2 process also used the corresponding reactivity of both processes to generate the remaining synergy values when excluding the contribution by reactive radicals. These findings provide a thorough understanding of the effects of UV and free chlorine on the degradation of ARGs and indicate the potential to utilize the combined processes of UV and free chlorine in water or wastewater treatment practice to control the dissemination of antibiotic resistance.

Assessment of the extent of bacterial growth in reverse osmosis system for improving drinking water quality.

This study was carried out to assess reverse osmosis (RO) treatment efficacy of drinking water in terms of biological stability in the distribution system. Two flat-sheet RO membranes were used in this study. Experiments were designed to investigate the growth of biofilm and bulk phase bacteria for the RO-treated water flowing through a model distribution system under controlled conditions without disinfectants. RO membranes improved the water quality of drinking water in terms of inorganic, organic and bacterial contents. Organic matter including the fraction available for microbes was efficiently removed by the RO membranes tested. More than 99% of bacterial cells in the tap water was retained by the RO membranes, leaving <50 cells/mL in the permeate water. In spite of the low nutrient contents and few cells in the RO permeates, monitoring of the model distribution systems receiving the RO permeates showed that remarkable biofilm accumulation and bulk cell growth occurred in the RO permeate water. In quasi-steady state, the total cell numbers in the biofilm and bulk water were of order 10(3) cells/cm(2) and 10(3) cells/mL, respectively, which were about 2 orders of magnitude lower than those grown in the tap water produced from conventional water treatment. The culturable heterotrophic bacteria constituted a significant part of the total cells (20.7-32.1% in biofilms and 21.3-46.3% in bulk waters). Biofilm maximum density and production rate were of the order 10(4) cells/cm(2) and 10(2) cells/cm(2)/day, respectively. The specific cell growth rate of bacteria in the biofilms was found to be much lower than those in the bulk waters (0.04-0.05 day(-1) versus 0.28-0.36 day(-1)). The overall specific cell growth rate which indicates the growth potential in the whole system was calculated as 0.07-0.08 day(-1), representing a doubling time of 9.1-10.1 days. These observations can be indicative of possibilities for bacterial growth in the RO permeate water with easily assimilable organic carbon concentrations below values proposed for biostability. RO permeate water does not appear to be biologically stable water. Therefore, efforts to minimize bacterial growth in the RO permeate water and in the distribution system must consider post-disinfection.

[Behaviors and environmental risk assessment of estrogenic endocrine disruptors in anaerobic domestic wastewater treatment].

Solid phase extraction-LC/MS/MS detection method was applied to study the concentrations and sludge adsorptions of nine estrogenic endocrine disruptors (EEDs) in an upflow anaerobic sludge blanket (UASB) and an anaerobic filter (AF). Nonylphenol equivalent quantity (NEQ) was calculated to evaluate the environmental risk of the EEDs. The results show anaerobic sludge adsorption has positive correlation with IgK(ow) and molecular length of EEDs. 17beta-estradiol (E2) and daidzein reached 83.2% and 90.4% removal rates in UASB, respectively. Genistein in AF is removed more than 80%. The sludge-water partition coefficient (K(p)) of nine EEDs ranged from 0.15-23.3 in UASB and 0.05-159.67 in AF. Biodegradation could be the main elimination pathway for daidzein, genistein, bisphenol A and estriol, by combining analysis of removal data and K(p) values. Environmental risk of EEDs was reduced after anaerobic treatment, as effluent NEQ was below the US EPA nonylphenol criteria (28 microg x L(-1), hour average concentration standard).

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.

Alternative immunofluorescent labeling of Cryptosporidium parvum in water samples using semiconductor quantum dots.

The application of immunofluorescent labeling using quantum dots for detection of inactivated Cryptosporidium parvum oocysts in spiked water samples (reservoir water, treated wastewater effluent, permeate of a membrane bioreactor, and tap water) provided more consistent results compared with the organic fluorophores label. The varying degree of particles present in the different water samples (with turbidity ranging from 0.2 to 6.1 NTU) in nonconcentrated water samples had insignificant interference on the labeled counts (2-sample t-tests, p > 0.236) using the quantum dot label, while the quantum dot label provided an advantage of approximately 50% lower interference in concentrated water samples compared with the organic fluorophores label.

Effect of formaldehyde on biofilm activity and morphology in an ultracompact biofilm reactor for carbonaceous wastewater treatment.

The effects of formaldehyde on biofilm morphology and biomass activity were investigated in an ultracompact biofilm reactor (UCBR) for carbonaceous wastewater treatment. The wastewater contained a fixed amount of glucose (with a chemical oxygen demand concentration of 600 mg/L) and an increasing concentration of formaldehyde (ranging from 21.4 to 271.1 mg/L). An influent formaldehyde concentration higher than 75 mg/L could facilitate filamentous growth (on biofilm) control and lead to a higher biofilm density, which is desirable as it enhanced the UCBR performance stability. However, at an influent formaldehyde concentration higher than 214.4 mg/L, biomass production was inhibited and deteriorations of biofilm morphology and biomass activity were observed. This study showed that it was desirable to maintain an influent formaldehyde concentration lower than 202.2 mg/L, as this concentration could achieve a good biofilm morphology while not inhibiting its microbial activity.

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.

Use of semiconductor quantum dots for photostable immunofluorescence labeling of Cryptosporidium parvum.

Cryptosporidium parvum is a waterborne pathogen that poses potential risk to drinking water consumers. The detection of Cryptosporidium oocysts, its transmissive stage, is used in the latest U.S. Environmental Protection Agency method 1622, which utilizes organic fluorophores such as fluorescein isothiocyanate (FITC) to label the oocysts by conjugation with anti-Cryptosporidium sp. monoclonal antibody (MAb). However, FITC exhibits low resistance to photodegradation. This property will inevitably limit the detection accuracy after a short period of continuous illumination. In view of this, the use of inorganic fluorophores, such as quantum dot (QD), which has a high photobleaching threshold, in place of the organic fluorophores could potentially enhance oocyst detection. In this study, QD605-streptavidin together with biotinylated MAb was used for C. parvum oocyst detection. The C. parvum oocyst detection sensitivity increased when the QD605-streptavidin concentration was increased from 5 to 15 nM and eventually leveled off at a saturation concentration of 20 nM and above. The minimum QD605-streptavidin saturation concentration for detecting up to 4,495 +/- 501 oocysts (mean +/- standard deviation) was determined to be 20 nM. The difference in the enumeration between 20 nM QD605-streptavidin with biotinylated MAb and FITC-MAb was insignificant (P > 0.126) when various C. parvum oocyst concentrations were used. The QD605 was highly photostable while the FITC intensity decreased to 19.5% +/- 5.6% of its initial intensity after 5 min of continuous illumination. The QD605-based technique was also shown to be sensitive for oocyst detection in reservoir water. This observation showed that the QD method developed in this study was able to provide a sensitive technique for detecting C. parvum oocysts with the advantage of having a high photobleaching threshold.

Microbial diversity and prevalence of virulent pathogens in biofilms developed in a water reclamation system.

Bacterial biofilm is a common phenomenon in both natural and engineered systems which often becomes a source of contamination and microbially influenced corrosion. It is thought that formation of biofilm in the monoculture of several bacterial species is regulated by acylhomoserine lactone (AHL) quorum-sensing signals. In this study, we investigated the microbial diversity and existence of AHL-producing and AHL-degrading bacterial species in the biofilm samples from a water reclamation system located in a tropical environment. 16S ribosomal DNA sequencing analysis indicated the presence of at least 11 bacterial species, including the frequently encountered bacterial pathogens Pseudomonas aeruginosa and Klebsiella pneumoniae, and several rare pathogens. We showed that only two groups of isolates, belonging to P. aeruginosa and Enterobacter agglomerans, produced AHL signals. We also found that three bacterial isolates, i.e., Agrobacterium tumefaciens XJ01, Bacillus cereus XJ08, and Ralstonia sp. XJ12, expressed AHL degradation enzymes. Furthermore, we showed that P. aeruginosa isolate HL43 was virulent against animal model Caenorhabditis elegans and released 2-6-fold more pyocyanin cytotoxin than P. aeruginosa strains PA01 and PA14, the two commonly used laboratory strains. These data indicate the complexity and importance of biofilm research in water reclamation.

A study on arsenic removal from household drinking water.

Arsenic removal from household drinking water has been receiving considerable attention in the field of water supply engineering. To develop the optimal coagulation protocol, the effectiveness of several operation options such as coagulants, coagulant aids and additives, as well as flocs separation systems were investigated in this study through the use of orthogonal array experiment based on Taguchi method. Arsenic removal mechanism during household coagulation (via manual mixing) was also discussed. The results showed that the addition of kaoline and powder activated carbon (PAC) did not enhance arsenic removal efficiency of ferric sulfate or aluminum sulfate. Similarly, mixture of ferric sulfate and aluminum sulfate (MFA) as well as polymeric ferric silicate sulfate (PFSiS) was also unable to improve the overall arsenic removal efficiency. The mechanism of arsenic removal during coagulation was somewhat different from those experienced in conventional processes. Coprecipitation appeared to be the crucial mechanism for arsenic removal. It is noted from this study that arsenic adsorption isotherm under household operation condition could be described by Langmuir equation. An efficient flocs separation system subsequent to coagulation was essential to achieve the effectiveness of overall arsenic removal. The results obtained from field experiment demonstrated that the method of ferric sulfate coagulation/sand filtration for arsenic removal from household drinking water was acceptable and affordable.

Effect of particles on the recovery of cryptosporidium oocysts from source water samples of various turbidities.

Cryptosporidium parvum can be found in both source and drinking water and has been reported to cause serious waterborne outbreaks which threaten public health safety. The U.S. Environmental Protection Agency has developed method 1622 for detection of Cryptosporidium oocysts present in water. Method 1622 involves four key processing steps: filtration, immunomagnetic separation (IMS), fluorescent-antibody (FA) staining, and microscopic evaluation. The individual performance of each of these four steps was evaluated in this study. We found that the levels of recovery of C. parvum oocysts at the IMS-FA and FA staining stages were high, averaging more than 95%. In contrast, the level of recovery declined significantly, to 14.4%, when the filtration step was incorporated with tap water as a spiking medium. This observation suggested that a significant fraction of C. parvum oocysts was lost during the filtration step. When C. parvum oocysts were spiked into reclaimed water, tap water, microfiltration filtrate, and reservoir water, the highest mean level of recovery of (85.0% +/- 5.2% [mean +/- standard deviation]) was obtained for the relatively turbid reservoir water. Further studies indicated that it was the suspended particles present in the reservoir water that contributed to the enhanced C. parvum oocyst recovery. The levels of C. parvum oocyst recovery from spiked reservoir water with different turbidities indicated that particle size and concentration could affect oocyst recovery. Similar observations were also made when silica particles of different sizes and masses were added to seeded tap water. The optimal particle size was determined to be in the range from 5 to 40 micro m, and the corresponding optimal concentration of suspended particles was 1.42 g for 10 liters of tap water.

Arsenic removal from household drinking water by adsorption.

Geogenic inorganic arsenic contamination in drinking water has been raising public health concern especially in developing countries. Cost-effective and stopgap arsenic removal method for household use (cooking and drinking) is very urgent. Several iron treated natural materials such as Fe-treated activated carbon (FeAC), Fe-treated gel beads (FeGB) and iron oxide-coated sand (IOCS), were investigated in this study for arsenic removal from dispersed household drinking water supply (scattered wells in the endemic arsenic poisoning areas). IOCS showed consistently good performance in terms of As(III) and As(V) removal in batch tests, column tests and field experiment. As(V) adsorption decreased slightly but As(III) adsorption maintained relatively stable when the pH value was increased from 5 to 9. In strong hardness water (612.5 mg/L CaCO3), As(III) adsorption efficiency was noted to decrease. The adsorption data obtained in column test fitted well to the Langmuir isotherm model. The adsorbent recovery efficiency was above 94% when using 0.2N NaOH regenerated the columns. In addition, 200 L of product water was produced by the household device (containing 3.0 kg IOCS produced) when the influent arsenic concentration ranging from 0.202 to 1.733 mg/L was encountered during the field experimental study conducted in Shanyin County, China. Neither the iron leaching nor other water quality deterioration was observed. It was noted in this study that IOCS is a promising medium for arsenic removal from household drinking water supplies.