Indicator Bacteria Concentrations Research Articles

Carbapenemase gene blaOXA-48 detected at six freshwater sites in Northern Ireland discharging onto identified bathing locations.

Faecal contamination of surface waters has the potential to spread not only pathogenic organisms but also antimicrobial resistant organisms. During the bathing season of 2021, weekly water samples, from six selected coastal bathing locations (n=93) and their freshwater tributaries (n=93), in Northern Ireland (UK), were examined for concentrations of faecal indicator bacteria Escherichia coli and intestinal enterococci. Microbial source tracking involved detection of genetic markers from the genus Bacteroides using PCR assays for the general AllBac marker, the human HF8 marker and the ruminant BacR marker for the detection of human, and ruminant sources of faecal contamination. The presence of beta-lactamase genes blaOXA-48, blaKPC, and blaNDM-1 was determined using PCR assays for the investigation of antimicrobial resistance genes that are responsible for lack of efficacy in major broad-spectrum antibiotics. The beta-lactamase gene blaOXA-48 was found in freshwater tributary samples at all six locations. blaOXA-48 was detected in 83% of samples that tested positive for the human marker and 69% of samples that tested positive for the ruminant marker over all six locations. This study suggests a risk of human exposure to antimicrobial resistant bacteria where bathing waters receive at least episodically substantial transfers from such tributaries.

Spatiotemporal assessment of pathogenic Leptospira in subtropical coastal watersheds.

The World Health Organization classifies leptospirosis as a significant public health concern, predominantly affecting impoverished and unsanitary regions. By using the Pensacola Bay System as a case study, this study examines the underappreciated susceptibility of developed subtropical coastal ecosystems such as the Pensacola Bay System to neglected zoonotic pathogens such as Leptospira. We analyzed 132 water samples collected over 12 months from 44 distinct locations with high levels of Escherichia coli (>410 most probable number/100 mL). Fecal indicator bacteria (FIB) concentrations were assessed using IDEXX Colilert-18 and Enterolert-18, and an analysis of water physiochemical characteristics and rainfall intensity was conducted. The LipL32 gene was used as a quantitative polymerase chain reaction (qPCR) indicator to identify the distribution of Leptospira interrogans. The results revealed 12 instances of the presence of L. interrogans at sites with high FIB over various land cover and aquatic ecosystem types. Independent of specific rainfall events, a seasonal relationship between precipitation and elevated rates of fecal bacteria and leptospirosis was found. These findings highlight qPCR's utility in identifying pathogens in aquatic environments and the widespread conditions where it can be found in natural and developed areas.

Seasonal dynamics and environmental drivers of tissue and mucus microbiomes in the staghorn coral Acropora pulchra.

Rainfall-induced coastal runoff represents an important environmental impact in near-shore coral reefs that may affect coral-associated bacterial microbiomes. Shifts in microbiome community composition and function can stress corals and ultimately cause mortality and reef declines. Impacts of environmental stress may be site specific and differ between coral microbiome compartments (e.g., tissue versus mucus). Coastal runoff and associated water pollution represent a major stressor for near-shore reef-ecosystems in Guam, Micronesia. Acropora pulchra colonies growing on the West Hagåtña reef flat in Guam were sampled over a period of 8 months spanning the 2021 wet and dry seasons. To examine bacterial microbiome diversity and composition, samples of A. pulchra tissue and mucus were collected during late April, early July, late September, and at the end of December. Samples were collected from populations in two different habitat zones, near the reef crest (farshore) and close to shore (nearshore). Seawater samples were collected during the same time period to evaluate microbiome dynamics of the waters surrounding coral colonies. Tissue, mucus, and seawater microbiomes were characterized using 16S DNA metabarcoding in conjunction with Illumina sequencing. In addition, water samples were collected to determine fecal indicator bacteria (FIB) concentrations as an indicator of water pollution. Water temperatures were recorded using data loggers and precipitation data obtained from a nearby rain gauge. The correlation structure of environmental parameters (temperature and rainfall), FIB concentrations, and A. pulchra microbiome diversity was evaluated using a structural equation model. Beta diversity analyses were used to investigate spatio-temporal trends of microbiome composition. Acropora pulchra microbiome diversity differed between tissues and mucus, with mucus microbiome diversity being similar to the surrounding seawater. Rainfall and associated fluctuations of FIB concentrations were correlated with changes in tissue and mucus microbiomes, indicating their role as drivers of A. pulchra microbiome diversity. A. pulchra tissue microbiome composition remained relatively stable throughout dry and wet seasons; tissues were dominated by Endozoicomonadaceae, coral endosymbionts and putative indicators of coral health. In nearshore A. pulchra tissue microbiomes, Simkaniaceae, putative obligate coral endosymbionts, were more abundant than in A. pulchra colonies growing near the reef crest (farshore). A. pulchra mucus microbiomes were more diverse during the wet season than the dry season, a distinction that was also associated with drastic shifts in microbiome composition. This study highlights the seasonal dynamics of coral microbiomes and demonstrates that microbiome diversity and composition may differ between coral tissues and the surface mucus layer.

Multiscale spatiotemporal variability of fecal indicator bacteria and associated particle size distributions in the sandy bottom sediments of a Pennsylvania creek.

Concentrations of the fecal indicator bacteria (FIB)Escherichia coliand enterococci are used to assess microbial impairment in irrigation and recreation water sources. Although the FIB concentrations' variability at large temporal scales, such as seasons, and large spatial scales encompassing different land use has been studied, the knowledge about smaller scale variability remains sparse. This work aimed to research the small-scale variability ofE. coliand enterococci in a montane creek with sandy bottom sediments. Sediment samples were collected weekly for a year in triplicate at sampling sites in aforested headwater, an agricultural area, and a mixed urban-agricultural area. The average weekly change in concentrations was from two times at the forested site to five times at the urban-agricultural site. Mean relative deviations from averages across sampling locations increased from -25% at the forested site to 45% at the urban-agricultural site. This trend was also observed separately over the cold and warm seasons. Over a week without precipitation, E. coliconcentrations decreased on average by 20% in warm period and by 45% in cold period; the enterococci concentration declined by 12% in both cold and warm periods. The sediment particle size distributions were significantly different among the three sites and between the cold and warm seasons. Rankings of sediment fine mass fractions and FIB concentrations were positively correlated at two of three sampling sites in more than 70% of observation dates. The results of this work indicate the need to evaluate the uncertainty of sediment FIB concentrations before designing sediment FIB monitoring quality.

Health impacts of poor water quality on an endangered shorebird breeding programme in Aotearoa New Zealand

ABSTRACT Case history Two clusters of mortality among endangered tūturuatu/tchūriwat’/shore plover (Thinornis novaeseelandiae) have occurred at captive breeding facilities around New Zealand in recent years. In the first, four chicks died at Pūkaha National Wildlife Centre (Mount Bruce, NZ) in February 2016, and in the second five adult birds at the Cape Sanctuary (Cape Kidnappers, NZ) died in 2022. Clinical findings In 2016, four chicks were noted to become weak, have increased vocalisations and closed eyes prior to death. The remaining chicks were treated for 5 days with amoxycillin/clavulanate orally twice daily. Water containers and brooders were cleaned and disinfected with chlorhexidine. No further mortality was seen. In the 2022 cluster, three adult breeding birds died acutely and five others showed inappetence, weight loss and diarrhoea approximately 10 days after heavy rains flooded the local river. The five birds were treated with amoxycillin/clavulanate orally twice daily and oral fluids for 5 days. Two birds died and three survived. No breeding occurred in the aviaries in the following season. Pathological findings In 2016, the chicks showed pulmonary changes ranging from congestion and oedema to heterophilic inflammation consistent with septicaemia. In 2022, the adult birds showed proliferation of bacteria in the distal small intestine associated with mucosal ulceration and heterophilic infiltration. Acid-fast staining of the caecal contents in one bird showed organisms consistent with Cryptosporidium spp. Laboratory findings Aerobic bacterial cultures of the lung and liver of two affected chicks carried out in 2016 showed heavy growth of Plesiomonas shigelloides. The same organism was cultured from water trays and holding tanks containing water boatmen (Sigara arguta) on which the chicks were fed. In 2022, cultures from the livers of three dead birds each showed a mixed bacterial growth with differing dominant organisms (Aeromonas sobria, Hafnia alvei, Citrobacter freundii and an Enterococcus sp.). PCR and sequencing confirmed Cryptosporidium parvum in the caecum of one bird. Fresh faeces from 24 breeding birds from the captive breeding facilities were negative by PCR for Cryptosporidium spp. The captive breeding facilities obtain water for the aviaries and aquatic invertebrates to feed to the chicks from local freshwater sources. Water quality testing at the Cape Sanctuary revealed concentrations of faecal indicator bacteria in excess of safe drinking water guidelines, with peaks following heavy rainfall. Clinical relevance Fluctuations in water quality associated with mammalian faecal bacteria can adversely affect bird health and impact on captive rearing of endangered wildlife.

Factors associated with foodborne pathogens and indicator organisms in agricultural soils

Soil can be a route for contamination of fresh fruits and vegetables. While growers routinely manage soil nutrient levels, little research exists on the synergistic or antagonistic effects of soil nutrients on foodborne pathogens. Data on foodborne pathogen prevalence in unamended soils is also relatively limited in literature. This study evaluated foodborne pathogen prevalence (Salmonella, Listeria monocytogenes) and concentration of indicator bacteria (total coliforms, generic Escherichia coli) in agricultural soils, and characterized associations between soil properties (e.g., macro- and micro-nutrient levels) and each microbial target. Three Virginia produce farms, representing different regions and soil types, were sampled four times over 1 year (October 2021–November 2022). For each individual farm visit, composite soil samples were collected from 20 sample sites (25 m2) per farm per visit for microbial and nutrient analysis (n = 240). Samples (25 g) were processed for Listeria spp. and Salmonella using a modified FDA BAM method; samples (5 g) were enumerated for generic E. coli and total coliforms (TC) using Petrifilm. Presumptive Listeria spp. and Salmonella isolates were confirmed by PCR using the sigB and invA genes, respectively. Soil nutrients from each sample were tested and evaluated for their association with each microbial target by Bayesian Mixed Models. Salmonella prevalence was 4.2% (10/240), with 90% (9/10) recovered on Farm C. Listeria spp. and L. monocytogenes prevalence were 10% (24/240) and 2.5% (6/240), respectively. When samples were positive for generic E. coli (107/240), the average concentration was 1.53 ± 0.77 log10 CFU/g. Soil pH was positively associated with L. monocytogenes [Odds Ratio (OR) = 5.5] and generic E. coli (OR = 4.9) prevalence. There was no association between Salmonella prevalence and any evaluated factor; however, Salmonella was 11.6 times more likely to be detected on Farm C, compared to other farms. Results show pathogen prevalence was relatively low in unamended soils, and that factors influencing prevalence and concentration varied by microbial target and farm.

Microbial hazards in real-world alternating dual-pit latrines treated with storage and lime in rural Cambodia

Abstract Achieving safely managed sanitation (SMS) in rural areas has spurred innovation in toilet designs that provide on-site treatment of fecal sludge (FS), including the development of International Development Enterprise (iDE)’s alternating dual-pit latrine upgrade (ADP). ADPs treat FS by inactivating pathogens using storage treatment with lime; however, ADPs’ reduction in pathogenicity (and thus their associated public health benefit) has not yet been described in real-world pits at scale. We thus enumerate the fecal indicator bacteria Escherichia coli (E. coli) and fecal coliforms in 147 pits after two years of storage treatment with lime and compare detected concentrations to relevant standards. E. coli and fecal coliform concentrations indicated a risk to human health in 31% and 42% of sampled pits, respectively. Regression models described relationships between fecal indicator bacteria concentrations and measured factors (e.g., sludge pH, temperature) but did not reveal any meaningful associations. High rates of pit ineligibility also indicate that many ADPs are not operated as recommended. Results indicate a one-in-three chance that a household emptying their own pit would be exposed to health hazards and call into question the effectiveness of the standard two-year storage treatment in real-world applications. To improve rural SMS, various evidence-based recommendations are made.

Monitoring Intertidal Habitats for Effects from Biosolids Applications onto an Adjacent Forestry Plantation

Stabilised organic solids derived from sewage sludge (“biosolids”) are applied to land as an alternative to disposal as landfill. This study evaluated the long-term effects of biosolids applied to forestry plantations on the adjacent intertidal habitats of Rabbit Island (New Zealand). On this island, biosolids are applied to enhance the growth of trees (Pinus radiata). Shoreline topography, macroalgal cover, sediment grain size, the concentrations of nutrients, trace metals, and faecal indicator bacteria, and benthic infaunal communities were studied in 2008, 2014, and 2019 at twelve intertidal transect sites (four “reference” and eight “application”) adjacent to forestry blocks where biosolids have been applied over a period of 24 years. The sediment composition did not differ significantly between the survey years or between the reference and application sites. Total nitrogen concentrations in the sediments increased over time at some transects, but such increases were not consistent among the application transects. No symptoms of excessive algal growth, sediment anoxia, and hydrogen sulphide odours were observed at most sites. Key infaunal taxa were similar between the reference and application transects. Overall, no long-term adverse changes to intertidal habitats attributed to biosolids application were detected between the reference and application sites. This study shows that biosolids application can co-occur without detectable adverse effects on nearby intertidal environments. In a global context of rising concern over climate change, environmental pollution, and resource scarcity, forest fertilisation with biosolids can facilitate biomass production and soil development while protecting valued coastal ecosystems.

Role of suspended solids on the co-precipitation of pathogenic indicators and antibiotic resistance genes with struvite from digested swine wastewater

Struvite recovered from wastewater contains high concentration of fecal indicator bacteria (FIB), porcine adenoviruses (PAdV) and antibiotic resistance genes (ARGs), becoming potential resources of these microbial hazards. Understanding the precipitation behavior of pathogenic indicators and ARGs with suspended solids (SS) will provide the possible strategy for the control of co-precipitation. In this study, SS was divided into high-density SS (separated by centrifugation) and low-density SS (further separated by filtration), and the role of SS on the co-precipitation of FIB, PAdV and ARGs was investigated. The distribution analysis showed that 35.5–73.0% FIB, 79.6% PAdV and 64.5–94.8% ARGs existed in high-density SS, while the corresponding values were 26.9–64.4%, 11.7% and 3.5–24.3% in low-density SS. During struvite generation, 82.7−96.9% FIB, 75.5% PAdV and 56.3–86.5% ARGs were co-precipitated into struvite. High-density SS contributed 20.7−68.5% FIB, 63.9% PAdV and 38.7–87.2% ARGs co-precipitation, and the corresponding contribution of low-density SS was 31.4−79.2%, 3.9% and 6.2–54.7%. Moreover, the precipitated SS in struvite obviously decreased inactivation efficiency of FIB and ARGs in drying process. These results provide a potential way to control the co-precipitation and inactivation of FIB, PAdV and ARGs in struvite through removing high-density SS prior to struvite recovery.

Health Risks to Communities and Athletes Associated with Swimming, Wading, and Sailing in Water Bodies of Brazil’s Guanabara Bay Basin

Guanabara Bay has been known to be polluted with trash and sewage from the surrounding areas, but health risks from recreational contact with water in the basin have not been well characterized. In this paper, fecal indicator bacteria (FIB) monitoring data are used to predict risks in three different exposure scenarios: (1) bathing in freshwater rivers that discharge into Guanabara Bay, (2) wading in these freshwater rivers, and (3) sailing in Guanabara Bay. Concentrations of indicator bacteria in river samples were measured directly, and concentrations of indicator bacteria in bay samples were sourced from publicly available government data sets. Ratios between pathogens and fecal indicator concentrations were used to estimate risks for five selected pathogens based on the indicator concentrations. The median risk of disease estimated from E. coli indicator concentrations was 1.0, 9.9 × 10−1 and 8.2 × 10−4 for the swimming, wading, and sailing exposure pathways, respectively. Risks estimates based on concentrations of the enterococci indicator bacteria in the sailing exposure scenario were comparable, at 3.4 × 10−4. The sum of total risk estimated from the five selected pathogens was 5.9 × 10−1, 3.6 × 10−1, and 1.0 × 10−3 for the swimming, wading, and sailing exposure pathways, respectively. Estimated risks of swimming and wading in the rivers far exceeded risks associated with U.S. recreational contact standards, while estimated risks for sailing in the bay were well below these risk guidelines. The 95th percentile of the sailing risk was estimated to exceed the U.S. recreational contact risk level. This paper exemplifies an approach to conducting quantitative microbial risk assessments when only fecal indicator bacteria data are available. Context-specific data on the relevant exposure routes, exposure frequency, and site-specific indicator: pathogen ratios were lacking, which ultimately led to uncertainty in the model. This study is intended to provide a framework for estimating GI risk based on fecal indicator concentrations while acknowledging that the substantial variation in indicator:pathogen ratios make the results of such efforts uncertain.

Treatment efficiency of package plants for on-site wastewater treatment in cold climates

Package plants (PP) are implemented around the world to provide on-site sanitation in areas not connected to a sewage network. The efficiency of PP has not been comprehensively studied at full scale, and the limited number of available studies have shown that their performance varies greatly. Their performance under cold climate conditions and the occurrence of micropollutants in PP effluents have not been sufficiently explored. PP are exposed to environmental factors such as low temperature, especially in cold regions with low winter temperatures and deep frost penetration, that can adversely influence the biochemical processes. The aim of this study was to investigate the treatment efficiency and possible effects of cold temperatures on PP performance, with focus on traditional contaminants (organics, solids, nutrients and indicator bacteria) and an additional assessment of micropollutants on two PP. Eleven PP hosting different treatment processes were monitored. Removal of biological oxygen demand (BOD) was high in all plants (>91%). Six out of the 11 PP provided good phosphorus removal (>71%). Small degrees of nitrification were observed in almost all the facilities, despite the low temperatures, while denitrification was only observed in two plants which achieved the highest nitrification rates (>51%) and had sludge recirculation. No strong correlation between wastewater temperature and BOD, nutrients and indicator bacteria concentration in the effluents was found. The high data variability and the effects of other process parameters as well as snow-melt water infiltration are suggested as possible reasons for the lack of correlation. However, weak negative relations between effluent concentrations and wastewater temperatures were detected in specific plants, indicating that temperature does have effects. When managed adequately, package plants can provide high BOD and phosphorus removal, but nitrogen and bacteria removal remain challenging, especially at low temperatures. Pharmaceutical compounds were detected in the effluents at concentrations within or above ranges reported for large treatment plants while phthalate ester concentrations were below commonly reported effluent concentrations.

Predicting Fecal Indicator Bacteria Using Spatial Stream Network Models in A Mixed-Land-Use Suburban Watershed in New Jersey, USA

Good water quality safeguards public health and provides economic benefits through recreational opportunities for people in urban and suburban environments. However, expanding impervious areas and poorly managed sanitary infrastructures result in elevated concentrations of fecal indicator bacteria and waterborne pathogens in adjacent waterways and increased waterborne illness risk. Watershed characteristics, such as urban land, are often associated with impaired microbial water quality. Within the proximity of the New York-New Jersey-Pennsylvania metropolitan area, the Musconetcong River has been listed in the Clean Water Act's 303 (d) List of Water Quality-Limited Waters due to high concentrations of fecal indicator bacteria (FIB). In this study, we aimed to apply spatial stream network (SSN) models to associate key land use variables with E. coli as an FIB in the suburban mixed-land-use Musconetcong River watershed in the northwestern New Jersey. The SSN models explicitly account for spatial autocorrelation in stream networks and have been widely utilized to identify watershed attributes linked to deteriorated water quality indicators. Surface water samples were collected from the five mainstem and six tributary sites along the middle section of the Musconetcong River from May to October 2018. The log10 geometric means of E. coli concentrations for all sampling dates and during storm events were derived as response variables for the SSN modeling, respectively. A nonspatial model based on an ordinary least square regression and two spatial models based on Euclidean and stream distance were constructed to incorporate four upstream watershed attributes as explanatory variables, including urban, pasture, forest, and wetland. The results indicate that upstream urban land was positively and significantly associated with the log10 geometric mean concentrations of E. coli for all sampling cases and during storm events, respectively (p < 0.05). Prediction of E. coli concentrations by SSN models identified potential hot spots prone to water quality deterioration. The results emphasize that anthropogenic sources were the main threats to microbial water quality in the suburban Musconetcong River watershed. The SSN modeling approaches from this study can serve as a novel microbial water quality modeling framework for other watersheds to identify key land use stressors to guide future urban and suburban water quality restoration directions in the USA and beyond.

Evaluation of physicochemical and microbiological parameters, and their correlation in Himalayan Spring Water Systems: A case study of District Kulgam of Kashmir Valley, India, Western Himalaya.

Total coliforms, E. coli, and fecal streptococci are the important indicators linked to the human health. This study investigated presence of these indicator bacteria in the Himalayan springs at various locations in the district Kulgam of Kashmir valley. A total of 30 spring water samples were collected from rural, urban, and forest areas during post-melting season 2021 and pre-melting season 2022. The springs in the area originate from the alluvium deposit, Karewa, and hard rock formations. The physicochemical parameters were found within the acceptable limits. However, nitrate and phosphate were found above the permissible limit at few sites, thus indicating the presence of anthropogenic activities in the area. Majority of the samples during both the seasons were found highly loaded with total coliforms with maximum limit of greater than 180 MPN/100ml. E. coli and fecal streptococci were found in the range of less than 1 to more than 180 MPN/100ml. The results of Pearson correlation of physicochemical parameters with indicator bacteria showed that chemical oxygen demand, rainfall, spring discharge, nitrate, and phosphate are the main factors affecting the concentration of indicator bacteria in the spring water at each site. Principal component analysis showed the most influencing factors of water quality in most of the spring sites are total coliforms, E. coli, fecal streptococci, rainfall, discharge, and chemical oxygen demand. The results of this study showed that the spring water is unfit for drinking purpose because of high concentration of fecal indicator bacteria.

Tidal effect on the dispersion of fecal pollution indicator bacteria and associated health risks along the Kribi beaches (Southern Atlantic coast, Cameroon)

Assessment of fecal contamination indicator bacteria concentrations on beaches are important for health risk prevention and management. This study assessed temporal changes in bacterial abundances as a function of tidal cycles and seasons along the Kribi beaches, Cameroon. Overall, samples taken during low and high tides during the different seasons of the year showed that the waters of Kribi beaches are exposed to fecal contamination due to the presence of different concentrations of fecal coliforms (5–35 CFU/100 ml), total coliforms (100–600 CFU/100 ml), Escherichia coli (0–15 CFU/100 ml), Streptococcus feacalis (50–700 CFU/100 ml), Pseudomonas aeruginosa (300–7200 CFU/100 ml), Vibrio cholerea (5–250 CFU/100 ml), Vibrio parahemoliticus (0–115 CFU/100 ml), and Aerobic Mesophilic Heterotrophic Bacteria (99–875 CFU/100 ml). Regardless of bacterial strain and sampling site, cell abundances were significant at low tides and during rainy seasons. At each sampling station (Mpalla, Ngoyè and Mboamanga), depending on the seasons and tidal cycles, significant correlations were recorded between the abundances of some bacterial strains and some environmental variables (P< 0.05). The presence and high abundance of these potentially pathogenic bacterial strains in the waters of Kribi beaches constitute a real public health threat for swimmers. The limitation of this microbiological pollution requires the implementation of an efficient collection and treatment plan for solid and liquid waste (sanitary wastewater) in the city of Kribi.

Prairie strips reduce fecal indicator bacteria concentrations in simulated runoff

AbstractVegetative filter strips (VFS) have shown promising results in reducing the downstream transport of many agroecosystem contaminants. A recently developed type of VFS, prairie strips, has been shown to significantly reduce the impact of corn and soybean production systems on water quality in terms of sediment, nitrogen, and phosphorus losses. This study assessed potential additional benefits of prairie strips to include the reduction of pathogens. To assess the impact of prairie strips on manure‐laden agricultural runoff, we utilized a physical model of prairie strips in a laboratory flume to conduct highly controlled overland flow experiments. Escherichia coli and Enterococcus concentration reductions of up to 45% and 65% were observed for runoff and infiltration flows, respectively, while mass load reductions of up to 65% were observed for surficial runoff flows. The degree of concentration or mass load reductions was dependent on the residence time of the flow within the strip and the partitioning of overland flow running onto the strip to infiltration and runoff flows. Based on our results and a review of the literature, we developed a design method to provide guidance on the width of prairie strip buffer needed to achieve a user‐defined reduction of fecal bacteria concentration.

A 22-Site Comparison of Land-Use Practices, E-coli and Enterococci Concentrations.

Land-use practices can greatly impact water quality. Escherichia (E.) coli and Enterococcus are accepted water quality indicators. However, surprisingly little research has been conducted comparing both organisms' population density relationships to land use practices and water quality. Stream water grab samples were collected monthly (n = 9 months) from 22 stream monitoring sites draining varying land use practice types in a representative mixed-land-use watershed of the northeastern United States. E. coli and enterococci colony forming units (CFU per 100 mL) were estimated (n = 396) and statistically analyzed relative to land use practices, hydroclimate, and pH, using a suite of methods, including correlation analysis, Principal Components Analysis (PCA), and Canonical Correspondence Analysis (CCA). Correlation analyses indicated significant (p < 0.05) relationships between fecal indicator bacteria concentrations, water quality metrics and land use practices but emphasized significant (p < 0.05) negative correlations between pH and instream enterococci concentrations. PCA and CCA results indicated consistent spatial differences between fecal indicator bacteria concentrations, pH, and land use/land cover characteristics. The study showed that pH could be considered an integrated proxy variable for past (legacy) and present land use practice influences. Results also bring to question the comparability of E-coli and enterococci relative to dominant land use practices and variations in pH and provide useful information that will help guide land use practice and water pollutant mitigation decision making.

Acanthamoeba and a bacterial endocytobiont isolated from recreational freshwaters.

The quality of many freshwater environments is impacted by human activities, so that many rivers may represent a vehicle for the transmission of health-related microorganisms. This work aimed to isolate and identify genetically free-living amoeba (FLA) of the genus Acanthamoeba from a recreational river in Salta, Argentina, and isolate, if possible, an endocytobiont. Sampling took place at four points (P1-P4) throughout the river in the winter and the summer seasons. Free-living amoebae and Acanthamoeba were recovered from 20-L water concentrated through an ultrafiltration system. Isolation was performed in agar plates, confirmation of Acanthamoeba genus by PCR, and fellow identification and classification based on their sequence analyses. High concentrations of indicator bacteria were found especially in P2, which is intensively used for recreation. Out of a total of 29 FLA isolations, 9 were identified as Acanthamoeba genotype T4 subtype A, the most frequent genotype found in nature and associated with causing human disease. From an axenic culture of Acanthamoeba spp. (KY751412), a bacterial endocytobiont was isolated, and identified as Stenotrophomonas maltophilia. The endocytobiont showed resistance and intermediate resistance to a wide range of widely used antibiotics. Results were in concordance with the cosmopolitan behavior of Acanthamoeba, and showed the importance of studying this group of amoebae and related microorganisms in recreational environments.

Implementing FAIR data management practices in shellfish sanitation

In the United States (U.S.), state agencies in charge of mariculture regulation are mandated under the U.S. Food and Drug Administration’s (FDA) National Shellfish Sanitation Program (NSSP) to monitor fecal indicator bacteria (FIB) concentrations, commonly of fecal coliforms, to determine the safety of coastal waters for supporting harvestable shellfish for human consumption. Many states have monitored bacteriological water quality for decades, creating impressive long-term records with the potential to advance foundational understanding of coastal systems and contribute to other complementary monitoring efforts. However, state shellfish sanitation programs differ in how they collect, manage, and share bacteriological monitoring data, resulting in their data typically being available in disparate state-level repositories with non-standardized database structures. Here, we outline three key recommendations as to how shellfish sanitation programs could implement practices to make their data more Findable, Accessible, Interoperable, and Reusable (FAIR), in turn creating new opportunities for the full potential of the data to be realized. We also offer sample materials of a standardized database, ShellBase, to provide an example of how diverse shellfish sanitation data may be integrated with a common data structure.

Total Maximum Daily Loads and Escherichia coli Trends in Texas Freshwater Streams

AbstractFecal indicator bacteria are routinely used to assess surface water sanitary quality. The State of Texas uses Total Maximum Daily Loads to address water bodies that exceed the allowable fecal indicator bacteria criteria. The effectiveness of these processes in decreasing the fecal indicator bacteria concentrations has been debated due to the diversity and nature of fecal indicator bacteria sources. We assessed actual and flow‐adjusted trends in measured Escherichia coli (E. coli) concentrations at 721 freshwater stream sites from 2001 through 2021. We also compared odds of statistical improvement of E. coli concentrations at sites before and after the adoption of Total Maximum Daily Loads (adopted from 2008 through 2014). Results indicate non‐significant differences in the odds of statistically detected improvements in E. coli concentration between pre‐Total Maximum Daily Load and post‐Total Maximum Daily Load sites. Although the State of Texas and numerous watershed stakeholders have made efforts to address water quality impairments, these results join a body of evidence that water quality improvements are stagnating in the state. Furthermore, this study leverages water quality data used for state water quality standards assessment purposes and highlights that robust monitoring program design is needed to effectively assess the progress of water quality planning efforts.

Tracking SARS-CoV-2 in rivers as a tool for epidemiological surveillance

The aim of this work was to evaluate if rivers could be used for SARS-CoV-2 surveillance. Five sampling points from three rivers (AR-1 and AR-2 in Arenales River, MR-1 and MR-2 in Mojotoro River, and CR in La Caldera River) from Salta (Argentina), two of them receiving discharges from wastewater plants (WWTP), were monitored from July to December 2020. Fifteen water samples from each point (75 in total) were collected and characterized physico-chemically and microbiologically and SARS-CoV-2 was quantified by RT-qPCR. Also, two targets linked to human contributions, human polyomavirus (HPyV) and RNase P, were quantified and used to normalize SARS-CoV-2 concentration, which was compared to reported COVID-19 cases. Statistical analyses allowed us to verify the correlation between SARS-CoV-2 and the concentration of fecal indicator bacteria (FIB), as well as to find similarities and differences between sampling points. La Caldera River showed the best water quality; FIBs were within acceptable limits for recreational activities. Mojotoro River's water quality was not affected by the northern WWTP of the city. Instead, Arenales River presented the poorest water quality; at AR-2 was negatively affected by the discharges of the southern WWTP, which contributed to significant increase of fecal contamination. SARS-CoV-2 was found in about half of samples in low concentrations in La Caldera and Mojotoro Rivers, while it was high and persistent in Arenales River. No human tracers were detected in CR, only HPyV was found in MR-1, MR-2 and AR-1, and both were quantified in AR-2. The experimental and normalized viral concentrations strongly correlated with reported COVID-19 cases; thus, Arenales River at AR-2 reflected the epidemiological situation of the city. This is the first study showing the dynamic of SARS-CoV-2 concentration in an urban river highly impacted by wastewater and proved that can be used for SARS-CoV-2 surveillance to support health authorities.