Please cite this article as: Purnell, S., Ebdon, J., Buck, A., Tupper, M., Taylor, H., Removal of phages and viral pathogens in a full-scale MBR: Implications for wastewater reuse and potable water, Water Research (2016),

The aim of this study was to assess the potential removal efficacy of enteric viruses in a fullscale membrane bioreactor (MBR) wastewater reuse system, using a range of indigenous and 'spiked' bacteriophages (phages) of known size and morphology. Samples were taken each week for three months from nine locations at each treatment stage of the water recycling plant (WRP) and tested for a range of microbiological parameters (n=135). Mean levels of faecal coliforms were reduced to 0.3 CFU/ 100ml in the MBR product and were undetected in samples taken after the chlorination stage. A relatively large reduction (5.3 log) in somatic coliphages was also observed following MBR treatment. However, F-RNA and humanspecific (GB124) phages were less abundant at all stages, and demonstrated log reductions post-MBR of 3.5 and 3.8, respectively. In 'spiking' experiments, free-swimming 'spiked' phages (MS2 and B14) displayed post-MBR log reductions of 2.25 and 2.30, respectively.The removal of these 'free-swimming' phages, which are smaller than the membrane pore size (0.04 µm), also highlights the possible role of the membrane biofilm as an effective additional barrier to virus transmission. The findings from this study of a full-scale MBR

This paper describes the isolation of Enterococcus host strains, for potential use as simple bacteriophage (phage)-based microbial source tracking (MST) tools. Presumptive Enterococcus host strains were isolated from cattle feces, raw municipal wastewater, agricultural runoff, and waters impacted by farms or wastewater treatment works (WWTW) in southern England, United Kingdom (UK). All enterococcal host strains (n = 390) were first screened for their ability to detect phage in samples of raw municipal wastewater and fecal material from cattle, pigs, and sheep. Host strains that detected phage (n = 147) were ranked according to both their specificity to a particular fecal source and also the number of phages (expressed as plaque-forming units, PFU) that they detected per milliliter of sample. Host strains that demonstrated host specificity and which detected phages at levels greater than 100 PFU/mL (n = 29) were further tested using additional fecal samples of human and nonhuman origin. The specificity and sensitivity of the enterococcal host strains were found to vary, ranging from 44 to 100% and from 17 to 83%, respectively. Most notably, seven strains exhibited 100% specificity to either cattle, human, or pig samples. Isolates exhibiting specificity to cattle were identified as belonging to the species Enterococcus casseliflavus , Enterococcus mundtii , or Enterococcus gallinarum , while human and pig isolates were members of either Enterococcus faecium or Enterococcus faecalis . The high specificity of phages infecting Enterococcus hosts and the simplicity and relatively low cost of the approach collectively indicate a strong potential for using this method as a tool in MST.

Water deficit, exacerbated by global population increases and climate change, necessitates the investigation of alternative non-traditional water sources to augment existing supplies.Indirect potable reuse (IPR) represents a promising alternative water source in water-stressed 2 regions. Of high concern is the presence of pathogenic microorganisms in wastewater, such as enteric viruses, protozoa and bacteria. Therefore, a greater understanding of the potential impact to human health is required. The aim of this research was to use a quantitative microbial risk assessment (QMRA) approach to calculate the probability of potential pathogen infection to the public in surface waters used for a range of recreational activities under scenarios: 1) existing de facto wastewater reuse conditions; 2) after augmentation with conventionally treated wastewater; and 3) after augmentation with reclaimed wastewater from proposed IPR schemes. Fourty-four 31 L samples were collected from river sites and a coastal wastewater treatment works from July 2016-May 2017. Concentrations of faecal indicator organisms (enterococci, faecal coliforms, somatic coliphages and Bacteroides phages) determined using culture-based approaches and selected pathogens (adenovirus, Salmonella and Cryptosporidium) determined using molecular approaches (qPCR) were used to inform QMRA. The mean probability of infection from adenovirus under de facto conditions was high (>0.90) for all recreational activities, per single event. The risk of adenovirus and Cryptosporidium infection increased under augmentation scenario (2) (mean probability 0.95-1.00 and 0.01-0.06 per single event, respectively). Adenovirus and Cryptosporidium infection risk decreased under reclaimed water augmentation scenario (3) (mean probability <0.79, excluding swimming, which remained 1.00 and <0.01 per single event, respectively). Pathogen reduction after reclaimed water augmentation in surface waters impacted by de facto reuse, provides important evidence for alternative water supply option selection. As such, this evidence may inform water managers and the public of the potential benefits of IPR and improve acceptance of such practices in the future.

Aim This study set out to determine whether phage‐based indicators may provide a ‘low‐tech’ alternative to existing approaches that might help maintain the microbial safety of shellfish and their overlying waters. Methods and Results Mussels and their overlying waters were collected biweekly from an estuary in southeast England over a 2‐year period (May 2013–April 2015) (n = 48). Levels of bacterial indicators were determined using membrane filtration and most probable number methods and those of bacteriophages were determined by direct plaque assay. The detection of adenovirus was determined using real‐time polymerase chain reaction. The results revealed that somatic coliphages demonstrated the most significant correlations with AdV F and G in mussels (ρ = 0·55) and overlying waters (ρ = 0·66), followed by GB124 phages (ρ = 0·43) while Escherichia coli showed no correlation with AdV F and G in mussels. Conclusion This study demonstrates that the use of somatic coliphages and GB124 phages may provide a better indication of the risk of adenovirus contamination of mussels and their overlying waters than existing bacterial indicators. Significance and Impact of the Study Phage‐based detection may be particularly advantageous in low‐resource settings where viral infectious disease presents a significant burden to human health.