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Objective: To assess the efficacy of a commercially available in‐water amphotericin B treatment for Macrorhabdus ornithogaster. Design Clinical treatment trial. Methods: Faecal shedding of 16 naturally infected budgerigars (Melopsittacus undulatus) was monitored while they were being treated using in‐water amphotericin B, as per the manufacturer's instructions, for 10 days. Any birds that remained positive after 10 days received a further 10 day course of treatment. All birds were rechecked 16 days after the end of the second treatment period. Results: At the conclusion of treatment, 11 birds had stopped shedding M. ornithogaster, and 5 birds were still shedding. Sixteen days after the conclusion of the second treatment period, four birds that were negative after 10 days of treatment were shedding again, and two of the birds that were treated for 20 days were shedding. In addition, one bird from each treatment group died after treatment and before follow‐up testing. Conclusion: These findings represent a 36% treatment failure, suggesting that treatment with the commercially available, water‐soluble amphotericin B has inconsistent efficacy against M. ornithogaster in some budgerigars in Australia and is not effective for eliminating it from budgerigar aviaries. [ABSTRACT FROM AUTHOR]

Objectives: Antimicrobial resistance (AMR) to critically important antimicrobials (CIAs) amongst Gram-negative bacteria can feasibly be transferred amongst wildlife, humans and domestic animals. This study investigated the ecology, epidemiology and origins of CIA-resistant Escherichia coli carried by Australian silver gulls (Chroicocephalus novaehollandiae), a gregarious avian wildlife species that is a common inhabitant of coastal areas with high levels of human contact.Methods: Sampling locations were widely dispersed around the perimeter of the Australian continent, with sites separated by up to 3500 km. WGS was used to study the diversity and molecular characteristics of resistant isolates to ascertain their epidemiological origin.Results: Investigation of 562 faecal samples revealed widespread occurrence of extended-spectrum cephalosporin-resistant (21.7%) and fluoroquinolone-resistant (23.8%) E. coli. Genome sequencing revealed that CIA-resistant E. coli isolates (n = 284) from gulls predominantly belonged to human-associated extra-intestinal pathogenic E. coli (ExPEC) clones, including ST131 (17%), ST10 (8%), ST1193 (6%), ST69 (5%) and ST38 (4%). Genomic analysis revealed that gulls carry pandemic ExPEC-ST131 clades (O25:H4 H30-R and H30-Rx) and globally emerging fluoroquinolone-resistant ST1193 identified among humans worldwide. Comparative analysis revealed that ST131 and ST1193 isolates from gulls overlapped extensively with human clinical isolates from Australia and overseas. The present study also detected single isolates of carbapenem-resistant E. coli (ST410-blaOXA-48) and colistin-resistant E. coli (ST345-mcr-1).Conclusions: The carriage of diverse CIA-resistant E. coli clones that strongly resemble pathogenic clones from humans suggests that gulls can act as ecological sponges indiscriminately accumulating and disseminating CIA-resistant bacteria over vast distances. [ABSTRACT FROM AUTHOR]

Microbial source tracking (MST) methods measure fecal contamination levels and identify possible sources using quantitative polymerase chain reaction (qPCR) that targets host-associated fecal microorganisms. To date, most established MST assays for human sources, especially bacterial markers, have shown some non-human host cross-reactions. Recently developed assays such as the crAssphage CPQ_056, Lachnospiraceae Lachno3 and Bacteroides BacV6-21 have more limited information on host sensitivity and host specificity for human or sewage sources, particularly in countries other than the USA. In this study, we rigorously evaluated six sewage-associated MST assays (i.e., Bacteroides HF183, human adenovirus (HAdV), human polyomavirus (HPyV), crAssphage CPQ_056, Lachno3 and BacV6-21) to show advantages and disadvantages of their applications for MST. A total of 29 human and 3 sewage samples, and 360 non-human fecal samples across 14 hosts collected from a subtropical region of Australia were tested for marker host specificity, host sensitivity and concentration. All sewage samples were positive for all six marker genes tested in this study. Bacterial markers were more prevalent in human feces than viral markers. Testing against animal hosts showed human feces (or sewage) associated marker gene specificity was HAdV (1.00) > HPyV (0.99) > crAssphage CPQ_056 (0.98) > HF183 (0.96) > Lachno3 (0.95) > BacV6-21 (0.90), with marker concentrations in some animal fecal samples being 3-5 orders of magnitude lower than in sewage. When considering host specificity, sensitivity, and concentrations in source samples, the HF183, Lachno3 and crAssphage CPQ_056 were the most suitable assays in this study for sewage contamination tracking in subtropical waters of Australia. [ABSTRACT FROM AUTHOR]

Lakes are one of the natural reservoirs of antibiotic resistance genes (ARGs) in environments. Long retention times in lakes potentially allow ARGs to persist and may create increased opportunities for the emergence of resistant pathogens. In this study, we investigated the prevalence, source and dissemination risk of ARGs in the sediments of a typical urban lake, Lake Tai (China) which has been a drastic example of water pollution with eutrophication in the world due to its proliferated cyanobacterial blooms. High-throughput profilings of ARGs in the sediments of Lake Tai were characterized with metagenomic assembly, and were compared with those in other global lakes from Australia, Canada, Indonesia, Rwanda and the United States of America. The hosts of ARGs in the sediments of Lake Tai were explored based on the taxonomic annotation of ARG-carrying contigs and network analysis, and a novel recently-discovered crAssphage was employed for source tracking of resistance bacteria. Meanwhile, the potential resistome risk was identified by projecting the co-occurrence of acquired ARGs, mobile genetic elements (MGEs) and human bacterial pathogens into a three-dimensional exposure space. Results showed 321 ARG subtypes belonging to 21 ARG types were detected in the sediments of Lake Tai, dominated by multidrug, macrolide-lincosamide-streptogramin, bacitracin, quinolone, mupirocin and trimethoprim resistance genes. Relatively, the ARG levels in the sediments of Lake Tai were significantly higher than those in other global lakes. Source tracking showed the coverages of detected crAssphage in the sediments of Lake Tai were positively correlated with the total ARG coverage, suggesting the contribution of human fecal contamination to the prevalence of ARGs in this lake. It should be noted that the co-occurrence ratio of ARGs, MGEs and human pathogens in the sediments of Lake Tai was higher than that in other global lakes, likely indicating a higher risk for the resistance dissemination in the China's third largest freshwater lake. • Prevalence and risk of ARGs in Lake Tai was studied by comparing with other global lakes. • CrAssphage was used for identifying the contribution of human fecal to the prevalence of ARGs. • Resistome risk was evaluated by considering the co-occurrence of ARGs, MGEs and HBPs. • ARG levels and resitome risk in Lake Tai were higher than those in other global lakes. [ABSTRACT FROM AUTHOR]

Recreational and potable water supplies polluted with human wastewater can pose a direct health risk to humans. Therefore, sensitive detection of human fecal pollution in environmental waters is very important to water quality authorities around the globe. Microbial source tracking (MST) utilizes human fecal markers (HFMs) to detect human wastewater pollution in environmental waters. The concentrations of these markers in raw wastewater are considered important because it is likely that a marker whose concentration is high in wastewater will be more frequently detected in polluted waters. In this study, quantitative PCR (qPCR) assays were used to determine the concentrations of fecal indicator bacteria (FIB) Escherichia coli and Enterococcus spp., HFMs Bacteroides HF183, human adenoviruses (HAdVs), and polyomaviruses (HPyVs) in raw municipal wastewater influent from various climatic zones in Australia. E. coli mean concentrations in pooled human wastewater data sets (from various climatic zones) were the highest (3.2×106 gene copies per ml), followed by those of HF183 (8.0×105 gene copies per ml) and Enterococcus spp. (3.6×105 gene copies per ml). HAdV and HPyV concentrations were 2 to 3 orders of magnitude lower than those of FIB and HF183. Strong positive and negative correlations were observed between the FIB and HFM concentrations within and across wastewater treatment plants (WWTPs). To identify the most sensitive marker of human fecal pollution, environmental water samples were seeded with raw human wastewater. The results from the seeding experiments indicated that Bacteroides HF183 was more sensitive for detecting human fecal pollution than HAdVs and HPyVs. Since the HF183 marker can occasionally be present in nontarget animal fecal samples, it is recommended that HF183 along with a viral marker (HAdVs or HPyVs) be used for tracking human fecal pollution in Australian environmental waters. [ABSTRACT FROM AUTHOR]

Estuaries are critical habitats subject to a range of stressors requiring effective management. Microbes are gaining recognition as effective environmental indicators, however, the response of host associated communities to stressors remains poorly understood. We examined microbial communities from seawater, sediments and the estuarine fish Pelates sexlineatus , in Australia's largest urbanised estuary, and hypothesised that anthropogenic contamination would be reflected in the microbiology of these sample types. The human faecal markers Lachno3 and HF183 were not detected, indicating negligible influence of sewage, but a gradient in copy numbers of the class 1 integron (intI-1), which is often used as a marker for anthropogenic contamination, was observed in sediments and positively correlated with metal concentrations. While seawater communities were not strongly driven by metal contamination, shifts in the diversity and composition of the fish gut microbiome were observed, with statistical links to levels of metal contamination (F 2, 21 = 1.536, p < 0.01). Within the fish gut microbiome, we further report increased relative abundance of amplicon sequence variants (ASVs; single inferred DNA sequences obtained in sequencing) identified as metal resistant and potentially pathogenic genera, as well as those that may have roles in inflammation. These results demonstrate that microbial communities from distinct habitats within estuarine systems have unique response to stressors, and alterations of the fish gut microbiome may have implications for the adaptation of estuarine fish to legacy metal contamination. [Display omitted] [ABSTRACT FROM AUTHOR]

Anthropogenic waste streams can be major sources of antibiotic resistant microbes within the environment, creating a potential risk to public health. We examined patterns in the occurrence of a suite of antibiotic resistance genes (ARGs) and their links to enteric bacteria at a popular swimming beach in Australia that experiences intermittent contamination by sewage, with potential points of input including stormwater drains and a coastal lagoon. Samples were collected throughout a significant rainfall event (40.8 mm over 3 days) and analysed using both qPCR and 16S rRNA amplicon sequencing. Before the rainfall event, low levels of faecal indicator bacteria and a microbial source tracking human faeces (sewage) marker (Lachno3) were observed. These levels increased over 10x following rainfall. Within lagoon, drain and seawater samples, levels of the ARGs sulI , dfrA1 and qnrS increased by between 1 and 2 orders of magnitude after 20.4 mm of rain, while levels of tetA increased by an order of magnitude after a total of 40.8 mm. After 40.8 mm of rain sulI , tetA and qnrS could be detected 300 m offshore with levels remaining high five days after the rain event. Highest levels of sewage markers and ARGs were observed adjacent to the lagoon (when opened) and in-front of the stormwater drains, pinpointing these as the points of ARG input. Significant positive correlations were observed between all ARGs, and a suite of Amplicon Sequence Variants that were identified as stormwater drain indicator taxa using 16S rRNA amplicon sequencing data. Of note, some stormwater drain indicator taxa, which exhibited correlations to ARG abundance, included the human pathogens Arcobacter butzleri and Bacteroides fragilis. Given that previous research has linked high levels of ARGs in recreationally used environments to antimicrobial resistant pathogen infections, the observed patterns indicate a potentially elevated human health risk at a popular swimming beach following significant rainfall events. [Display omitted] • Levels of human sewage markers were elevated during rainfall. • Levels of ARGs correlated significantly with markers for human sewage. • Not all ARGS displayed the same spatiotemporal distribution patterns. • ARGs significantly correlated with stormwater drain indicator bacteria. • Some ARGs were correlated with known human pathogens. [ABSTRACT FROM AUTHOR]

The World Health Organization recommends that the majority of water monitoring laboratories in the world test for E. coli daily since thermotolerant coliforms and E. coli are key indicators for risk assessment of recreational waters. Recently, we developed a new SNP method for typing E. coli strains, by which human-specific genotypes were identified. Here, we report the presence of these previously described specific SNP profiles in environmental water, sourced from the Coomera River, located in South East Queensland, Australia, over a period of two years. This study tested for the presence of human-specific E. coli to ascertain whether hydrologic and anthropogenic activity plays a key role in the pollution of the investigated watershed or whether the pollution is from other sources. We found six human-specific SNP profiles and one animal-specific SNP profile consistently across sampling sites and times. We have demonstrated that our SNP genotyping method is able to rapidly identify and characterize human- and animal-specific E. coli isolates in water sources. [ABSTRACT FROM AUTHOR]

The article discusses the study which evaluates the specificity of JC and BK polyomaviruses through polymerase chain reaction (PCR) wastewater testing and fecal sampling from various animals in Queensland, Australia. It implies that researchers have developed microbial source tracking tools to distinguish human from animal fecal pollution in environmental waters to minimize public health risk of exposure to pathogens. The study indicates that JC and BK viruses that are specific to human wastewater are detected in a coastal river which proves the presence of human fecal pollution.

This paper reports a reconnaissance survey of the concentrations of sterol compounds (as indicators of fecal contamination) in a large water supply system in southeast Australia comprising a network of rivers, channels, and drains. Levels of coprostanol and cholestanol were determined in surface water and bottom sediment using gas chromatography–mass spectrometry analysis across 17 strategic sampling sites and over 12 months. Clear differences in the levels of fecal contamination were observed among sites. Four sites routinely contained high levels of the fecal indicator sterols indicated from surface water and sediment sample analysis. Coprostanol concentrations at each location varied from 0 ng/L at the reference site to 11,327 ng/L in a surface water sample of a drain directly downstream of a knackery. The majority of the sites contained coprostanol in the range of 500 to 800 ng/L. Since no fecal-associated sterol compounds were detected at the external reference sites, these were assumed to be free from fecal contamination. Sewage water discharge and/or substantial water runoff maybe the principal factors contributing to fecal contamination of the supply drains and channels. [ABSTRACT FROM AUTHOR]

Increasing urbanisation and changes in land use lead to adverse impacts on the quality of natural water resources. The specific sources of contamination are often difficult to identify using conventional water quality monitoring techniques. This acts as a significant constraint to the development of appropriate management techniques to protect natural water resources. Consequently, alternative means of identifying pollutant sources and their locality are necessary. In this study, Antibiotic Resistance Patterns (ARP) were established for a library of 1005 known Escherichia coli source isolates obtained from human and non-human (domesticated animals, livestock and wild) sources in an urbanising catchment in Queensland State, Australia. Discriminant Analysis (DA) was used to differentiate between the ARP of source isolates and to identify the sources of faecal contamination. Partial Least Square (PLS) regression was then utilised on identified human source isolates to correlate their locality with specified sampling locations within the catchment. The resulting ARP DA indicated that a majority of the faecal contamination in the rural areas was non-human. However, the percentage of human isolates increased significantly in urbanised areas using onsite systems for wastewater treatment. The PLS regression was able to develop predictive models which indicated a high correlation of human source isolates from the urban area. The study results confirm the feasibility of using ARP for source tracking faecal contamination in surface waters, as well as predicting their point of origin. [Copyright &y& Elsevier]

Aims: The aim of this study was to identify the prevalence of environmentally adapted enterococci strains by analysing biochemical fingerprinting (BF) data of 3952 enterococci isolates collected over 5 years from the six catchments in Southeast Queensland, Australia. Methods and Results: A BF method was used to type 3952 enterococci isolates from six catchments. The environmental isolates were compared with a large existing BF library comprised of 5803 enterococci isolates from 10 host groups. Environmental isolates belonged to 801 biochemical phenotypes (BPTs), of which, an average of 29.2% was specific to each catchment. When compared with the BF library, an average of 79·5% BPTs from each catchment was identical to those in the library (i.e. host-origin BPTs). The remaining 20·5% was regarded as non-host origin BPTs, as they were not in the library and constituted only 5·3% of the total isolates tested for each catchment. Conclusions: Our data suggest that less than 5% of studied environmental strains was not identical to those in the library and seemed to be of environmental origin. From a microbial source tracking context, such low level of environmentally adapted strains can have a minimal impact on the performance of the library-based methods if a large number of isolates were tested from both the host groups and environmental waters. Significance and Impact of the Study: These data shed light on the importance of the size and representativeness of library–based source-tracking methods and their implications for the identification of faecal pollution in environmental waters. [ABSTRACT FROM AUTHOR]

This study aimed to determine the prevalence and abundance of sewage and animal fecal contamination of sediment at seven estuarine locations in Sydney, NSW, Australia. Sediment samples were tested for the occurrence of microbial targets including molecular marker genes of enterococci (ENT), Bacteroides HF183 (HF183), Methanobrevibacter smithii (nifH) , human adenovirus (HAdV) and emerging sewage-associated marker genes crAssphage (CPQ_056) and Lachnospiraceae (Lachno3) and animal feces-associated marker genes, including avian feces-associated Helicobacter spp. (GFD), canine-feces associated Bacteroides (DogBact), cattle-feces associated (cowM2) and horse feces-associated Bacteroides (HoF597). Results from this study showed that urban estuarine sediment can act as a reservoir of fecal indicator bacteria (FIB) and several microbial source tracking (MST) marker genes, including previously unreported Lachno3. The sewage-associated marker gene CPQ_056 was most prevalent, in 63.8% of sediment samples, while the avian associated marker gene GFD had the highest mean abundance. The GFD marker gene was highly abundant and widely detected in sediment samples from all seven locations compared to the other animal feces-associated marker genes. In all, 31 (44.9%) sediment samples were positive for at least two sewage-associated marker genes. However, the non-quantifiable detection of the HAdV marker gene did not always align with the detection of two or more sewage-associated marker genes. In addition, the most frequent wet weather overflow exposure occurred at locations that did not have a consistent pattern of detection of the sewage-associated marker genes, suggesting sediments may not be a suitable measure of recent sewage contamination. To assist water quality and public health managers better understand past microbial contamination of estuarine sediment, further studies seem justified to explore the role of decay of MST marker genes in sediment. Further work is also needed on the role of resuspension of MST marker genes from sediment during storm events to the water column as a source of contamination for both the GFD and sewage-associated marker genes. • The crAssphage marker gene was detected in 63.8% of sediment samples. • The avian marker was highly abundant and widely detected in sediment. • Adenovirus did not always align with the detection of two or more sewage markers. • The prevalence of the sewage marker genes did not align well with wet weather overflows. [ABSTRACT FROM AUTHOR]

Rapid and widespread monitoring of inland and coastal water quality occurs through the use of remote sensing and near-surface water quality sensors. A new addition is the development of smartphone applications (Apps) to measure and record surface reflectance, water color and water quality parameters. In this paper, we present a field study of the HydroColor (HC, measures RGB reflectance and suspended particulate matter (SPM)) and EyeOnWater (EoW, determines the Forel–Ule scale—an indication to the visual appearance of the water surface) smartphone Apps to evaluate water quality for inland waters in Eastern Australia. The Brisbane river, multiple lakes and reservoirs and lagoons in Queensland and New South Wales were visited; hyperspectral reflection spectra were collected and water samples were analysed in the laboratory as reference. Based on detailed measurements at 32 sites, covering inland waters with a large range in sediment and algal concentrations, we find that both water quality Apps are close, but not quite on par with scientific spectrometers. EoW is a robust application that manages to capture the color of water with accuracy and precision. HC has great potential, but is influenced by errors in the observational procedure and errors in the processing of images in the iPhone. The results show that repeated observations help to reduce the effects of outliers, while implementation of camera response functions and processing should help to reduce systematic errors. For both Apps, no universal conversion to water quality composition is established, and we conclude that: (1) replicated measurements are useful; (2) color is a reliable monitoring parameter in its own right but it should not be used for other water quality variables, and; (3) tailored algorithms to convert reflectance and color to composition could be developed for lakes individually. [ABSTRACT FROM AUTHOR]

Animal fecal contamination in aquatic environments is a major source of zoonotic diseases in humans. While concerns are focused on livestock, companion animals such as dogs can also be a source of a wide range of zoonotic pathogens. Therefore, detection of dog or canine fecal contamination in aquatic environments is important for mitigating risks. In this study, host-sensitivity and specificity of four canine fecal-associated marker genes were evaluated by analyzing 30 canine and 240 non-canine fecal samples. The application of these markers was also tested in water from an urban river under dry weather conditions. The host sensitivity values of the Bacteroides BacCan-UCD, DogBact, DF113 and DF418 were 1.00, 0.90, 0.83, and 0.90, respectively. The host specificity value of the BacCan-UCD, DogBact, DF113 and DF418 were 0.87, 0.98, 0.83, and 0.41, respectively. The mean concentrations of DF418 were highest (7.82 ± 1.13 log 10 gene copies (GC)/g of feces) followed by BacCan-UCD (7.61 ± 1.06 log 10 GC/g) and DogBact (7.15 ± 0.92 log 10 GC/g). The mean concentration of DF113 (5.80 ± 1.25 log 10 GC/g) was 1.5 to 2.5 orders of magnitude lower than the other marker genes. The DogBact marker gene was not detected in any other animal feces other than a small number of untreated sewage samples. The BacCan-UCD marker gene cross-reacted with cat, chicken, and pig fecal samples, while the DF113 marker gene cross-reacted with cat, chicken, cattle fecal and untreated sewage samples. The DF418 marker gene was detected in all sewage and animal feces and deemed not suitable for canine fecal contamination tracking in sub-tropical Australia. Canine fecal contamination was infrequently detected in environmental water samples. Based on the results obtained in this study, we recommend that at least two canine feces-associated marker genes should be used in field studies., Competing Interests: Declaration of competing interest The authors declare no conflict of interest, (Crown Copyright © 2020. Published by Elsevier B.V. All rights reserved.)

Characterization of microbial communities using high-throughput amplicon sequencing is an emerging approach for microbial source tracking of fecal pollution. This study used SourceTracker software to examine temporal and geographical variability of fecal bacterial community profiles to identify pollutant sources in three freshwater catchments in sub-tropical Australia. Fecal bacterial communities from 10 animal species, humans, and composite wastewater samples from six sewage treatment plants were characterized and compared to freshwater samples using Illumina amplicon sequencing of the V5–V6 regions of the 16S rRNA gene. Source contributions were calculated in SourceTracker using new fecal taxon libraries as well as previously generated libraries to determine the effects of geographic and temporal variability on source assignments. SourceTracker determined 16S rRNA bacterial communites within freshwater samples, shared taxonomic similarities to that of wastewater at low levels (typically <3%). SourceTraker also predicted occasional fecal detection of deer and flying fox sources in the water samples. No significant differences in source contributions were observed within sequences from current and previously characterized fecal samples (P ≥ 0.107). However, significant differences were observed between previously characterized and newly characterized source communities (ANOSIM P ≤ 0.001), which shared <15% community composition. Results suggest temporal instability of fecal taxon libraries among tested sources and highlight continual evaluation of community-based MST using confirmatory qPCR analyses of marker genes. Image 1 • 16S rRNA sequencing established microbial profiles of three catchments. • Sewage microbial communities showed low similarity to those of water samples. • Deer and flying fox fecal contamination were found in several samples. • Dog fecal contamination showed a greater contribution than did other sources. • Dog fecal taxon libraries showed the greatest temporal stability over the other source libraries examined. [ABSTRACT FROM AUTHOR]

Bacteria monitoring is a critical part of wastewater management. At tropical wastewater stabilization ponds (WSPs) in north Australia, sanitation is assessed using the standard fecal indicator bacteria (FIB) Escherichia coli and Enterococci. However, these bacteria are poor surrogates for enteric pathogens. A focus on FIB misses the majority of pond-bacteria and how they respond to the tropical environment. Therefore, we aimed to identify the unknown pond bacteria and indicators that can complement E. coli to improve monitoring. Over two years, we measured the bacterial community in 288 wastewater samples during the wet and dry seasons. The WSP community was spatially and temporally dynamic. Standard pond-water physicochemical measures like conductivity poorly explained these community shifts. Cyanobacteria represented >6% of the WSP bacterial population, regardless of sample timing and location. Fecal bacteria were abundant in the first pond. However, in downstream ponds, these bacteria were less abundant, and instead, environmental taxa were common. For each pond, we identified a bacterial fingerprint that included new candidate bacterial indicators of fecal waste and processes like nitrogen removal. Combining the new indicators with standard FIB monitoring represents a locally relevant approach to wastewater monitoring that facilitates new tests for human fecal pollution within tropical climates. [ABSTRACT FROM AUTHOR]

Background: Biofilters are soil-plant based passive stormwater treatment systems which demonstrate promising, although inconsistent, removal of faecal microorganisms. Antimicrobial-producing plants represent a safe, inexpensive yet under-researched biofilter design component that may enhance treatment reliability. The mechanisms underlying plant-mediated microbial removal in biofilters have not been fully elucidated, particularly with respect to antimicrobial production. The aim of this study was therefore to inform biofilter vegetation selection guidelines for optimal pathogen treatment by conducting antimicrobial screening of biofilter-suitable plant species. This involved: (1) selecting native plants suitable for biofilters (17 species) in a Victorian context (southeast Australia); and (2) conducting antimicrobial susceptibility testing of selected plant methanolic extracts (≥ 5 biological replicates/species; 86 total) against reference stormwater faecal bacteria (Salmonella enterica subsp. enterica ser. Typhimurium, Enterococcus faecalis and Escherichia coli). Results: The present study represents the first report on the inhibitory activity of polar alcoholic extracts from multiple tested species. Extracts of plants in the Myrtaceae family, reputed for their production of antimicrobial oils, demonstrated significantly lower minimum inhibitory concentrations (MICs) than non-myrtaceous candidates (p < 0.0001). Melaleuca fulgens (median MIC: 8 mg/mL; range: [4–16 mg/mL]), Callistemon viminalis (16 mg/mL, [2–16 mg/mL]) and Leptospermum lanigerum (8 mg/mL, [4–16 mg/mL]) exhibited the strongest inhibitory activity against the selected bacteria (p < 0.05 compared to each tested non-myrtaceous candidate). In contrast, the Australian biofilter gold standard Carex appressa demonstrated eight-fold lower activity than the highest performer M. fulgens (64 mg/mL, [32–64 mg/mL]). Conclusion: Our results suggest that myrtaceous plants, particularly M. fulgens, may be more effective than the current vegetation gold standard in mediating antibiosis and thus improving pathogen treatment within biofilters. Further investigation of these plants in biofilter contexts is recommended to refine biofilter vegetation selection guidelines. [ABSTRACT FROM AUTHOR]

Aquatic recreation in urban estuaries worldwide is often restricted by fecal pollution. Variability in the occurrence of fecal pathogens and their differential virulence potentials within these estuaries may result in variable public health risks. To address this hypothesis, Campylobacter were isolated from the Yarra River estuary, Australia and then characterized via HeLa cell cytotoxicity and attachment to and the invasion of Caco-2 monolayers. Overall, 54% (n=216) of estuarine samples (water and sediment combined) yielded biochemically confirmed culturable Campylobacter; higher detection was recorded in water (92%, n=90) than in the bank and bed sediments combined (27%, n=126). The seasonality of occurrence was not significant. HeLa cell cytotoxicity revealed that estuarine Campylobacter had low cytotoxin titers; the 95% confidence interval (CI) ranged between 61 and 85, which was markedly lower than the mean value (~386) for the C. jejuni 11168 reference pathogenic strain. The Caco-2 attachment of estuarine Campylobacter isolates (n=189) revealed that the 95%CI for the attachment efficiency of the test strains ranged between 0.09 and 0.1%, with only 3.7% having a higher efficiency than the 5 th percentile value for C. jejuni 11168. None of the estuarine strains exhibited Caco-2 invasion capabilities. In contrast to the common assumption during quantitative microbial/risk assessments (QMRAs) that all environmental strains are pathogenic, the present results revealed that Campylobacter within the Yarra River estuary had very low virulence potential. Since this is the first study to use human epithelial cell lines to characterize estuary-borne pathogens, these results generate valuable insights for a better understanding of the public health risks in urban estuaries that will underpin more robust QMRAs.