Comparison of kits for SARS-CoV-2 extraction in liquid and passive samples.

Effective extraction and detection of viral nucleic acids from sewage are fundamental components of a successful SARS-CoV-2 sewage surveillance programme. As there is no standard method employed in sewage surveillance, understanding the performance of different extraction kits in the recovery of SARS-CoV-2 and the impact that PCR inhibitors have on quantification is essential to minimize data discrepancies caused by sample extraction. Three commercial nucleic acid extraction kits: the RNeasy PowerSoil Total RNA Kit (PS), the RNeasy PowerMicrobiome Kit (PMB), and the MagMAX™ Microbiome Ultra Nucleic Acid Isolation Kit (MM), with minor modifications, were evaluated.

The detection of Japanese encephalitis virus in municipal wastewater during an acute disease outbreak.

Aim: To demonstrate the capability of wastewater-based surveillance (WBS) as a tool for detecting potential cases of Japanese Encephalitis Virus (JEV) infection in the community.

Methods And Results: In this study, we explore the potential of WBS to detect cases of JEV infection by leveraging from an established SARS-CoV-2 wastewater surveillance program. We describe the use of two reverse transcriptase quantitative polymerase chain reaction (RTqPCR) assays targeting JEV to screen archived samples from two wastewater treatment plants (WWTPs).

Recycling drinking water treatment sludge into eco-concrete blocks with CO curing: Durability and leachability.

Drinking water treatment sludge (DWTS) can be recycled into low-strength concrete blocks for construction use. The sodium sulfate resistance and leaching behaviours of the DWTS-derived blocks are investigated in this study. The experimental results show that the addition of DWTS degrades the sodium sulfate resistance of the concrete blocks, however CO curing compensates for such property, especially in the case of blocks incorporating 30% DWTS.

Utilization of drinking water treatment sludge in concrete paving blocks: Microstructural analysis, durability and leaching properties.

The management of abundant drinking water treatment sludge (DWTS) in landfill remains an important issue. The reuse of DWTS as construction material could contribute to the development of greener concrete product and to mitigating the detrimental environment effect from excessive production of DWTS. This paper investigates the potential of using DWTS as sand replacement in Concrete Paving Blocks (CPB).

Chlorine inactivation of coxsackievirus B5 in recycled water destined for non-potable reuse.

Currently guidelines for disinfection of water with free chlorine, while primarily developed for potable water, are often used for virus disinfection of nitrified recycled water of >1 NTU (Nephelometric Turbidity Unit). More information is needed on the disinfection efficacy of free chlorine for viruses in waters of varying turbidity and pH due to significant reuse of treated wastewater of varying quality. In this study, disinfection efficacy in nitrified/denitrified activated sludge treated wastewater was investigated for coxsackievirus B5 (CB5), an enterovirus known to be highly resistant to free chlorine.

Virus removal by ultrafiltration: Understanding long-term performance change by application of Bayesian analysis.

Ultrafiltration is an effective barrier to waterborne pathogens including viruses. Challenge testing is commonly used to test the inherent reliability of such systems. Performance validation seeks to demonstrate the adequate reliability of the treatment system.

Bayesian belief network modelling of chlorine disinfection for human pathogenic viruses in municipal wastewater.

Chlorine disinfection of biologically treated wastewater is practiced in many locations prior to environmental discharge or beneficial reuse. The effectiveness of chlorine disinfection processes may be influenced by several factors, such as pH, temperature, ionic strength, organic carbon concentration, and suspended solids. We investigated the use of Bayesian multilayer perceptron (BMLP) models as efficient and practical tools for compiling and analysing free chlorine and monochloramine virus disinfection performance as a multivariate problem.

Modelling pathogen log10 reduction values achieved by activated sludge treatment using naïve and semi naïve Bayes network models.

Risk management for wastewater treatment and reuse have led to growing interest in understanding and optimising pathogen reduction during biological treatment processes. However, modelling pathogen reduction is often limited by poor characterization of the relationships between variables and incomplete knowledge of removal mechanisms. The aim of this paper was to assess the applicability of Bayesian belief network models to represent associations between pathogen reduction, and operating conditions and monitoring parameters and predict AS performance.

Uncertainties associated with assessing the public health risk from Legionella.

Legionella is an opportunistic pathogen of public health concern. Current regulatory and management guidelines for the control of this organism are informed by risk assessments. However, there are many unanswered questions and uncertainties regarding Legionella epidemiology, strain infectivity, infectious dose, and detection methods.

Detection of Legionella, L. pneumophila and Mycobacterium avium complex (MAC) along potable water distribution pipelines.

Inhalation of potable water presents a potential route of exposure to opportunistic pathogens and hence warrants significant public health concern. This study used qPCR to detect opportunistic pathogens Legionella spp., L.

Fate of pathogenic microorganisms and indicators in secondary activated sludge wastewater treatment plants.

This study was undertaken to investigate the removal of pathogenic microorganisms and their indicators in a laboratory scale biological treatment system that simulated the secondary treatment process of a wastewater treatment plant (WWTP). Four groups of microorganisms including bacteria, viruses, protozoa and helminths as well as the selected indicators were employed in the investigation. The results demonstrated that approximately 2-3 log10 removal of the microbial indicators was achieved in the treatment process.

Effect of water treatment processes on Cryptosporidium infectivity.

Conventional water treatment processes have the ability to remove Cryptosporidium oocysts through coagulation, flocculation, sedimentation and filtration, provided there is efficient management of plant performance. The potential exists for the breakthrough of oocysts through the treatment train. The effect of the water treatment chemical aluminium sulphate (alum) on Cryptosporidium oocyst infectivity has been assessed using an assay that combines cell culture and real-time polymerase chain reaction techniques.

Bacterial degradation of microcystin toxins within a biologically active sand filter.

Microcystin toxins are a problem for water authorities as they are recalcitrant to conventional water treatment. In this study, biological sand filtration was assessed in laboratory column experiments for its ability to remove two microcystin analogues, microcystin-LR and microcystin-LA. A lag period of 3 days was evident prior to the commencement of degradation.

Environmental temperature controls Cryptosporidium oocyst metabolic rate and associated retention of infectivity.

Cryptosporidium is a significant cause of water-borne enteric disease throughout the world and represents a challenge to the water industry and a threat to public health. In this study we report the use of a cell culture-TaqMan PCR assay to measure oocyst inactivation rates in reagent-grade and environmental waters over a range of temperatures. While oocysts incubated at 4 degrees C and 15 degrees C remained infective over the 12-week holding period, we observed a 4 log(10) reduction in infectivity for both 20 and 25 degrees C incubation treatments at 12 and 8 weeks, respectively, for all water types examined, a faster rate of inactivation for oocysts than previously reported.

Emerging technologies for the detection and genetic characterization of protozoan parasites.

The development and adaptation of new technologies for the genetic characterization and identification of parasites continue to accelerate, providing an increasing number of research and analytical tools. We review emerging technologies that have applications in this area, including real-time PCR and microarrays, and discuss the fundamental principles of some of these technologies and how they are applied to characterize parasites. We give special consideration to the application of genetic data to biological questions, where selection of the most appropriate technique depends on the biological question posed by the investigator.

Cell culture-Taqman PCR assay for evaluation of Cryptosporidium parvum disinfection.

Cryptosporidium parvum represents a challenge to the water industry and a threat to public health. In this study, we developed a cell culture-quantitative PCR assay to evaluate the inactivation of C. parvum with disinfectants.