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Legionella is the causative agent of Legionnaires’ disease, and its prevalence in potable water is a significant public health issue. Water stagnation within buildings increases the risk of Legionella. However, there are limited studies investigating how stagnation arising through intermittent usage affects Legionella proliferation and the studies that are available do not consider viable but non culturable (VBNC) Legionella. This study used a model plumbing system to examine how intermittent water stagnation affects both VBNC and culturable Legionella. The model plumbing system contained a water tank supplying two biofilm reactors. The model was initially left stagnant for ≈5 months (147 days), after which one reactor was flushed daily, and the other weekly. Biofilm coupons, and water samples were collected for analysis at days 0, 14 and 28. These samples were analysed for culturable and VBNC Legionella, free-living amoebae, and heterotrophic bacteria. After 28 days, once-a-day flushing significantly (p

AbstractPresumptive tests or lateral flow immunoassay testing kits are increasingly being utilised as a fast, portable, cost-effective method to detect an array of analytes, particularly illicit drugs. In Australia, the methamphetamine and decontamination industry regularly use presumptive testing kits to determine whether a property is contaminated. These presumptive test kits advertise a detection limit of 0.5 µg/100 cm2 and above for methamphetamine. The performance of seven methamphetamine presumptive test brands and their kit components were analysed in this study. Following the kit instructions, each kit was analysed using a ceramic tile that was spiked with a known concentration of methamphetamine. These presumptive test kits were highly sensitive, with positive results also found below the limit of detection. Therefore, if there is a negative result it is likely that there is no methamphetamine present. Those that return a positive result require further quantitative assessment. Thus, the presumptive test kits may detect the presence of methamphetamine that is below the Australian Clandestine Drug Laboratory Remediation Guideline of 0.5 µg/100 cm2. This indicates these tests might be useful as a screening assessment but should not be relied upon to develop a remediation action plan.

Airborne microorganisms play a significant role in the transmission of infectious diseases. As such, improving indoor microbial air quality can enhance infection control in numerous settings. This study examined the efficacy of the PlasmaShield® air purification device to remove airborne microorganisms under laboratory conditions. Pure cultures of model microorganisms at varying concentrations were aerosolized using a 1-jet Collison nebulizer through stainless-steel removable piping prior to reaching the PlasmaShield® device. The surviving microorganisms were captured using the Staplex® MBS-6 Six Stage Microbial Air Sampler and enumerated via culture on agar plates. The positive-hole-corrected colony/plaque-forming units were compared with the negative control (microorganisms aerosolized through an empty PlasmaShield® casing). The PlasmaShield® statistically significantly (p < 0.05) reduced airborne Escherichia coli, Staphylococcus epidermidis, Bacteriophage MS2 and Cladosporium sp. compared with the negative control. The maximum removal achieved was estimated to be 4 × log10E. coli (99.99% removal), 4 × log10S. epidermidis (99.97% removal), 7 × log10 MS2 (99.99998% removal) and 5 × log10Cladosporium sp. (99.999% removal). Scanning electron microscope images of the surviving microorganisms showed that the PlasmaShield® damaged the cell membrane of these model microorganisms. This study provides proof-of-concept evidence to support the use of this technology to improve indoor microbial air quality.

Legionella pneumophila is a waterborne pathogen and, as the causative agent of Legionnaires’ disease, a significant public health concern. Exposure to environmental stresses, and disinfection treatments, promotes the formation of resistant and potentially infectious viable but non-culturable (VBNC) Legionella. The management of engineered water systems to prevent Legionnaires’ disease is hindered by the presence of VBNC Legionella that cannot be detected using the standard culture (ISO11731:2017-05) and quantitative polymerase reaction (ISO/TS12869:2019) methods. This study describes a novel method to quantify VBNC Legionella from environmental water samples using a “viability based flow cytometry-cell sorting and qPCR” (VFC + qPCR) assay. This protocol was then validated by quantifying the VBNC Legionella genomic load from hospital water samples. The VBNC cells were unable to be cultured on Buffered Charcoal Yeast Extract (BCYE) agar; however, their viability was confirmed through their ATP activity and ability to infect amoeba hosts. Subsequently, an assessment of the ISO11731:2017-05 pre-treatment procedure demonstrated that acid or heat treatment cause underestimation of alive Legionella population. Our results showed that these pre-treatment procedures induce culturable cells to enter a VBNC state. This may explain the observed insensitivity and lack of reproducibility often observed with the Legionella culture method. This study represents the first time that flow cytometry-cell sorting in conjunction with a qPCR assay has been used as a rapid and direct method to quantify VBNC Legionella from environmental sources. This will significantly improve future research evaluating Legionella risk management approaches for the control of Legionnaires’ disease.

Methamphetamine manufacture, use, and the resulting contamination is a significant issue that affects public health, the environment, and the economy. Third-hand exposure to methamphetamine can result in adverse health risks for individuals and first responders. Such exposures can result from the inhalation of airborne residues or from contact with contaminated objects. This review was conducted to determine the current methods used for methamphetamine extraction from indoor air and porous fabric materials. Dynamic solid phase microextraction (SPME) and sorbent sampling tubes have been applied to extract airborne methamphetamine residues from contaminated properties. SPME and solvent extraction have been applied to sample clothing and textiles for methamphetamine detection. This review demonstrates that there is limited literature on the detection of methamphetamine from indoor air and clothing. Supplementary and consistent methods to detect methamphetamine from air and porous surfaces should be developed and published to allow better assessment of the environmental risk to public health caused by third-hand exposure to methamphetamine.

Legionella is an opportunistic waterborne pathogen associated with Legionnaires' disease and Pontiac fever. Despite improved public awareness, the incidence of Legionella associated infections has been increasing. Aerosols generated from engineered potable water systems are a demonstrated cause of both nosocomial and community-acquired legionellosis. The ecology of Legionella in these systems is complex with multiple factors impacting their colonization and persistence. Flow dynamics has been identified as an important factor and stagnation in cooling towers is an accepted risk for increased Legionella growth; however, less is known about the impact of flow dynamic on Legionella in potable water systems. This is especially complex due to the inherent intermittent and variable usage observed within outlets of a potable water system. This systematic literature review examines the role of fluid dynamics and stagnation on the colonization and growth of Legionella in potable water systems. Twenty two of 24 identified studies show a positive association between stagnation zones and increased colonization of Legionella. These zones included dead legs, dead ends, storage tanks, and obstructed water flow (such as intermittent usage or flow restriction). Prolonged stagnation in building plumbing systems also deteriorates the quality of thermally or chemically treated potable water. This stimulates the colonization of Legionella established biofilms. Such biofilms are intrinsically resistant to disinfection procedures and accelerate the rate of decay of chemical disinfectants. Sub-lethal doses of disinfectants and the presence of protozoan hosts in stationary water promote generation of viable but non-culturable Legionella cells. This results in false negatives in surveillance methods that use culture methodology. In conclusion, elimination of temporal and permanent stagnation points can improve the quality of potable water, efficacy of disinfectants, and reduce the risk of legionellosis. Current guidelines and water safety plans recognize the risks associated with permanent stagnation point (dead ends and dead legs); however, there is a need for greater emphasis on controlling temporal stagnation arising from intermittent usage.

In response to the Coronavirus Disease 2019 (COVID-19) pandemic, current modeling supports the use of masks in community settings to reduce the transmission of SARS-CoV-2. However, concerns have been raised regarding the global shortage of medical grade masks and the limited evidence on the efficacy of fabric masks. This study used a standard mask testing method (ASTM F2101-14) and a model virus (bacteriophage MS2) to test the viral filtration efficiency (VFE) of fabric masks compared with commercially available disposable, surgical, and N95 masks. Five different types of fabric masks were purchased from the ecommerce website Etsy to represent a range of different fabric mask designs and materials currently available. One mask included a pocket for a filter; which was tested without a filter, with a dried baby wipe, and a section of a vacuum cleaner bag. A sixth fabric mask was also made according to the Victorian Department of Health and Human Services (DHHS) guidelines (Australia). Three masks of each type were tested. This study found that all the fabric masks had a VFE of at least 50% when tested against aerosols with an average size of 6.0 µm (VFE(6.0 µm)). The minimum VFE of fabric masks improved (to 63%) when the larger aerosols were excluded to give and average aerosol size of 2.6 µm (VFE(2.6 µm)), which better represents inhaled aerosols that can reach the lower respiratory system. The best performing fabric masks were the cotton mask with a section of vacuum cleaner bag (VFE(6.0 µm) = 99.5%, VFE(2.6 µm) = 98.8%) or a dried baby wipe (VFE(6.0 µm) = 98.5%, VFE(2.6 µm) = 97.6%) in the pocket designed for a disposable filter, the mask made using the Victorian DHHS design (VFE(6.0 µm) = 98.6%, VFE(2.6 µm) =99.1%) and one made from a layer of 100% hemp, a layer of poly membrane, and a layer of cheesecloth (VFE(6.0 µm) = 93.6%, VFE(2.6 µm) = 89.0%). The VFE of two surgical masks (VFE(6.0 µm) = 99.9% and 99.6%, VFE(2.6 µm) = 99.5% and 98.5%) and a N95 masks (VFE(6.0 µm) = 99.9%, VFE(2.6 µm) = 99.3%) were comparable to their advertised bacterial filtration efficacy. This research supports the use of fabric masks in the community to prevent the spread of SARS-CoV-2; however, future research is needed to explore the optimum design in ensuring proper fit. There is also a need for mass education campaigns to disseminate this information, along with guidelines around the proper usage and washing of fabric masks.

Campylobacter spp. is one of the most widespread infectious diseases of veterinary and public health significance. Globally, the incidence of campylobacteriosis has increased over the last decade in both developing and developed countries. Squamates (lizards, snakes and amphisbaenians) are a potential reservoir and source of transmission of campylobacteriosis to humans. This systematic review examined studies from the last 20 years that have reported squamate-associated human campylobacteriosis. It was found that C. fetus subsp. testudinum and C. fetus subsp. fetus were the most common species responsible for human campylobacteriosis from a squamate host. The common squamate hosts identified included bearded dragons (Pogona vitticeps), green iguana (Iguana iguana), western beaked gecko (Rhynchoedura ornate) and blotched blue-tongued skink (Tiliqua nigrolutea). People with underlying chronic illnesses, the immunocompromised and the elderly were identified as the most vulnerable population. Exposure to pet squamates, wild animals, consumption of reptilian cuisines and cross contamination with untreated water were risk factors associated with Campylobacter infections. Proper hand hygiene practices, responsible pet ownership, ‘One Health’ education and awareness on zoonotic diseases will help reduce the public health risks arising from Campylobacter exposure through squamates. Continued surveillance using molecular diagnostic methods will also enhance detection and response to squamate-linked campylobacteriosis.

Healthcare-associated infections (HAIs) are one of the most common patient complications, affecting 7% of patients in developed countries each year. The rise of antimicrobial resistant (AMR) bacteria has been identified as one of the biggest global health challenges, resulting in an estimated 23,000 deaths in the US annually. Environmental reservoirs for AMR bacteria such as bed rails, light switches and doorknobs have been identified in the past and addressed with infection prevention guidelines. However, water and water-related devices are often overlooked as potential sources of HAI outbreaks. This systematic review examines the role of water and water-related devices in the transmission of AMR bacteria responsible for HAIs, discussing common waterborne devices, pathogens, and surveillance strategies. AMR strains of previously described waterborne pathogens including Pseudomonas aeruginosa, Mycobacterium spp., and Legionella spp. were commonly isolated. However, methicillin-resistant Staphylococcus aureus and carbapenem-resistant Enterobacteriaceae that are not typically associated with water were also isolated. Biofilms were identified as a hot spot for the dissemination of genes responsible for survival functions. A limitation identified was a lack of consistency between environmental screening scope, isolation methodology, and antimicrobial resistance characterization. Broad universal environmental surveillance guidelines must be developed and adopted to monitor AMR pathogens, allowing prediction of future threats before waterborne infection outbreaks occur.

Legionella pneumophila is an opportunistic waterborne pathogen of public health concern. It is the causative agent of Legionnaires’ disease (LD) and Pontiac fever and is ubiquitous in manufactured water systems, where protozoan hosts and complex microbial communities provide protection from disinfection procedures. This review collates the literature describing interactions between L. pneumophila and protozoan hosts in hospital and municipal potable water distribution systems. The effectiveness of currently available water disinfection protocols to control L. pneumophila and its protozoan hosts is explored. The studies identified in this systematic literature review demonstrated the failure of common disinfection procedures to achieve long term elimination of L. pneumophila and protozoan hosts from potable water. It has been demonstrated that protozoan hosts facilitate the intracellular replication and packaging of viable L. pneumophila in infectious vesicles; whereas, cyst-forming protozoans provide protection from prolonged environmental stress. Disinfection procedures and protozoan hosts also facilitate biogenesis of viable but non-culturable (VBNC) L. pneumophila which have been shown to be highly resistant to many water disinfection protocols. In conclusion, a better understanding of L. pneumophila-protozoan interactions and the structure of complex microbial biofilms is required for the improved management of L. pneumophila and the prevention of LD.

Strongyloides spp. are parasitic nematodes that are transmitted through the environment and are capable of causing disease. These nematodes affect an estimated 3−300 million humans worldwide. Identifying the environmental reservoirs of Strongyloides spp. is essential for the development of appropriate control strategies. This systematic literature review examined all published studies that identified Strongyloides stercoralis, Strongyloides fuelleborni, Strongyloides fuelleborni kellyi, and Strongyloides spp. from an environmental source. Most studies detected the nematode from dog and primate fecal samples. Other environmental sources identified were ruminants, cats, rodents, insects, water, soil, as well as fruit and vegetables. Most studies used microscopy-based identification techniques; however, several employed molecular-based techniques, which have become increasingly popular for the detection of Strongyloides spp. A limitation identified was a lack of studies that comprehensively screened all potential environmental samples in a region. Future research should undertake this holistic screening process to identify which environmental reservoirs pose the greatest significance to human health. Potential controls can be identified through the identification of environmental sources. Understanding where Strongyloides spp. is commonly found within the environment of endemic areas will inform environmental control strategies to reduce this neglected disease.

In Australia, rainwater is an important source of water for many households. Unlike municipal water, rainwater is often consumed untreated. This study investigated the potential contamination of rainwater by microorganisms. Samples from 53 rainwater tanks across the Adelaide region were collected and tested using Colilert™ IDEXX Quanti-Tray*/2000. Twenty-eight out of the 53 tanks (53%) contained Escherichia coli. Samples collected from ten tanks contained E. coli at concentrations exceeding the limit of 150 MPN/100 mL for recreational water quality. A decline in E. coli was observed in samples collected after prolonged dry periods. Rainwater microbiological values depended on the harvesting environment conditions. A relationship was found between mounted TV antenna on rooftops and hanging canopies; and E. coli abundance. Conversely, there was no relationship between seasonality and E. coli or roof and tank structure materials and E. coli. In several tanks used for drinking water, samples collected prior to and after filtration showed that the filtration systems were not always successful at completely removing E. coli. These results differed from a study undertaken in the laboratory that found that a commercially available in-bench 0.45 µm filter cartridge successfully reduced E. coli in rainwater to 0 MPN/100 mL. After running a total of 265 L of rainwater which contained high levels of E. coli through the filter (half of the advertised filter lifespan), the filter cartridge became blocked, although E. coli remained undetected in filtered water. The difference between the laboratory study and field samples could be due to improper maintenance or installation of filters or recontamination of the faucet after filtration. The presence of E. coli in water that is currently used for drinking poses a potential health concern and indicates the potential for contamination with other waterborne pathogens.