Humans create vast quantities of wastewater through inefficiencies and poor management of water systems. The wasting of water poses sustainability challenges, depletes energy reserves, and undermines human water security and ecosystem health. Here we review emerging approaches for reusing wastewater and minimizing its generation. These complementary options make the most of scarce freshwater resources, serve the varying water needs of both developed and developing countries, and confer a variety of environmental benefits. Their widespread adoption will require changing how freshwater is sourced, used, managed, and priced.

A nested-PCR method was used to detect the occurrence of human adenovirus in coastal waters of Southern California. Twenty- to forty-liter water samples were collected from 12 beach locations from Malibu to the border of Mexico between February and March 1999. All sampling sites were located at mouths of major rivers and creeks. Two ultrafiltration concentration methods, tangential flow filtration (TFF) and vortex flow filtration (VFF), were compared using six environmental samples. Human adenoviruses were detected in 4 of the 12 samples tested after nucleic acid extraction of VFF concentrates. The most probable number of adenoviral genomes ranged from 880 to 7,500 per liter of water. Coliphages were detected at all sites, with the concentration varying from 5.3 to 3332 PFU/liter of water. F-specific coliphages were found at 5 of the 12 sites, with the concentration ranging from 5.5 to 300 PFU/liter. The presence of human adenovirus was not significantly correlated with the concentration of coliphage ( r = 0.32) but was significantly correlated ( r = 0.99) with F-specific coliphage. The bacterial indicators (total coliforms, fecal coliforms, and enterococci) were found to exceed California recreational water quality daily limits at 5 of the 12 sites. However, this excess of bacterial indicators did not correlate with the presence of human adenoviruses in coastal waters. The results of this study call for both a reevaluation of our current recreational water quality standards to reflect the viral quality of recreational waters and monitoring of recreational waters for human viruses on a regular basis.

A total of 26 strains of Vibrio cholerae , including members of the O1, O139, and non-O1, non-O139 serogroups from both clinical and environmental sources, were examined for the presence of genes encoding cholera toxin ( ctx A), zonula occludens toxin ( zot ), accessory cholera enterotoxin ( ace ), hemolysin ( hly A), NAG-specific heat-stable toxin ( st ), toxin-coregulated pilus ( tcp A), and outer membrane protein ( omp U), for genomic organization, and for the presence of the regulatory protein genes tcp I and tox R in order to determine relationships between epidemic serotypes and sources of isolation. While 22 of the 26 strains were hemolytic on 5% sheep blood nutrient agar, all strains were PCR positive for hly A, the hemolysin gene. When multiplex PCR was used, all serogroup O1 and O139 strains were positive for tcp A, omp U, and tcp I. All O1 and O139 strains except one O1 strain and one O139 strain were positive for the ctx A, zot , and ace genes. Also, O1 strain VO3 was negative for the zot gene. All of the non-O1, non-O139 strains were negative for the ctx A, zot , ace , tcpA , and tcp I genes, and all of the non-O1, non-O139 strains except strain VO26 were negative for omp U. All of the strains except non-O1, non-O139 strain VO22 were PCR positive for the gene encoding the central regulatory protein, tox R. All V. cholerae strains were negative for the NAG-specific st gene. Of the nine non- ctx -producing strains of V. cholerae , only one, non-O1, non-O139 strain VO24, caused fluid accumulation in the rabbit ileal loop assay. The other eight strains, including an O1 strain, an O139 strain, and six non-O1, non-O139 strains, regardless of the source of isolation, caused fluid accumulation after two to five serial passages through the rabbit gut. Culture filtrates of all non-cholera-toxigenic strains grown in AKI media also caused fluid accumulation, suggesting that a new toxin was produced in AKI medium by these strains. Studies of clonality performed by using enterobacterial repetitive intergenic consensus sequence PCR, Box element PCR, amplified fragment length polymorphism (AFLP), and pulsed-field gel electrophoresis (PFGE) collectively indicated that the V. cholerae O1 and O139 strains had a clonal origin, whereas the non-O1, non-O139 strains belonged to different clones. The clinical isolates closely resembled environmental isolates in their genomic patterns. Overall, there was an excellent correlation among the results of the PCR, AFLP, and PFGE analyses, and individual strains derived from clinical and environmental sources produced similar fingerprint patterns. From the results of this study, we concluded that the non-cholera-toxin-producing strains of V. cholerae , whether of clinical or environmental origin, possess the ability to produce a new secretogenic toxin that is entirely different from the toxin produced by toxigenic V. cholerae O1 and O139 strains. We also concluded that the aquatic environment is a reservoir for V. cholerae O1, O139, non-O1, and non-O139 serogroup strains.

The paucity of proper sanitation facilities has contributed to the spread of waterborne diseases in many developing countries. The primary goal of this study was to demonstrate the feasibility of using a wastewater electrolysis cell (WEC) for toilet wastewater disinfection. The treated wastewater was designed to reuse for toilet flushing and agricultural irrigation. Laboratory-scale electrochemical (EC) disinfection experiments were performed to investigate the disinfection efficiency of the WEC with four seeded microorganisms (Escherichia coli, Enterococcus, recombinant adenovirus serotype 5, and bacteriophage MS2). In addition, the formation of organic disinfection byproducts (DBPs) trihalomethanes (THMs) and haloacetic acids (HAA5) at the end of the EC treatment was also investigated. The results showed that at an applied cell voltage of +4 V, the WEC achieved 5-log10 reductions of all four seeded microorganisms in real toilet wastewater within 60 min. In contrast, chemical chlorination (CC) disinfection using hypochlorite [NaClO] was only effective for the inactivation of bacteria. Due to the rapid formation of chloramines, less than 0.5-log10 reduction of MS2 was observed in toilet wastewater even at the highest [NaClO] dosage (36 mg/L, as Cl2) over a 1 h reaction. Experiments using laboratory model waters showed that free reactive chlorine generated in situ during EC disinfection process was the main disinfectant responsible for the inactivation of microorganisms. However, the production of hydroxyl radicals [OH], and other reactive oxygen species by the active bismuth-doped TiO2 anode were negligible under the same electrolytic conditions. The formation of THMs and HAA5 were found to increase with higher applied cell voltage. Based on the energy consumption estimates, the WEC system can be operated using solar energy stored in a DC battery as the sole power source.

Pathogenic bacteria and enteric viruses can be introduced into the environment via human waste discharge. Methods for rapid detection and quantification of human viruses and fecal indicator bacteria in water are urgently needed to prevent human exposure to pathogens through drinking and recreational waters. Here we describe the development of two real-time PCR methods to detect and quantify human adenoviruses and enterococci in environmental waters. For real-time quantification of enterococci, a set of primers and a probe targeting the 23S rRNA gene were used. The standard curve generated using Enterococcus faecalis genomic DNA was linear over a 7-log-dilution series. Serial dilutions of E. faecalis suspensions resulted in a lower limit of detection (LLD) of 5 CFU/reaction. To develop real-time PCR for adenoviruses, degenerate primers and a Taqman probe targeting a 163-bp region of the adenovirus hexon gene were designed to specifically amplify 14 different serotypes of human adenoviruses, including enteric adenovirus serotype 40 and 41. The standard curve generated was linear over a 5-log-dilution series, and the LLD was 100 PFU/reaction using serial dilutions of purified adenoviral particles of serotype 40. Both methods were optimized to be applicable to environmental samples. The real-time PCR methods showed a greater sensitivity in detection of adenoviruses in sewage samples than the viral plaque assay and in detection of enterococci in coastal waters than the bacterial culture method. However, enterococcus real-time PCR overestimated the number of bacteria in chlorinated sewage in comparison with the bacterial culture method. Overall, the ability via real-time PCR to detect enterococci and adenoviruses rapidly and quantitatively in the various environmental samples represents a considerable advancement and a great potential for environmental applications.