Contrasting Water Quality Treatments Result in Structural and Functional Changes to Wetland Plant-Associated Microbial Communities in Lab-Scale Mesocosms.

The impact of contrasting water quality treatments on wetland plant-associated microbial communities was investigated in this study using 12 lab-scale wetland mesocosms (subsurface flow design) planted with reed canary grass (Phalaris arundinacea) or water speedwell (Veronica anagallis-aquatica) over a 13-week period. Mesocosms received water collected from two sites along the Grand River (Ontario, Canada) designated as having either high or poor water quality according to Grand River Conservation Authority classifications. All mesocosms were established using sediment collected from the high water quality site and received water from this source pre-treatment. Resulting changes to microbial community structure were assessed using PCR-denaturing gel gradient electrophoresis (DGGE) on microbial 16S rDNA sequences extracted from rhizoplane, rhizosphere, and water samples before and after exposure to water quality treatments. Functional community changes were determined using Biolog™ EcoPlates which assess community-level carbon source utilization profiles. Wetland mesocosm removal of inorganic nutrients (N, P) and fecal coliforms was also determined, and compared among treatments. Treatment-specific effects were assessed using a repeated measures restricted maximum likelihood (REML) analysis. Structural and functional characteristics of rhizoplane microbial communities were significantly influenced by the interaction between plant species and water treatment (P = 0.04, P = 0.01). Plant species-specific effects were observed for rhizosphere structural diversity (P = 0.01) and wetland water community metabolic diversity (P = 0.03). The effect of water treatment alone was significant for structural diversity measurements in wetland water communities (P = 0.03). The effect of plant species, water quality treatment, and the interaction between the two is dependent on the microhabitat type (rhizoplane, rhizosphere, or water). Rhizoplane communities appear to be more sensitive to water quality-specific environmental changes and may be a good candidate for microbial community-based monitoring of wetland ecosystems.

Pilot-scale comparison of sodium silicates, orthophosphate and pH adjustment to reduce lead release from lead service lines.

Sodium silicate is thought to mitigate lead release via two mechanisms: by increasing pH and by forming a protective silica film. A pilot-scale study using an excavated lead service line (LSL) fed with water from a Great Lakes source was undertaken to: (1) clearly distinguish the pH effect and the silica effect; (2) compare sodium silicate to orthophosphate and pH adjustment; (3) determine the nature of silica accumulation in the pipe scale. The LSL was cut into segments and acclimated with water at pH 7.1. Median dissolved lead was 197 µg/L in the last 8 weeks of acclimation and dropped to 16 µg/L, 54 µg/L, and 85 µg/L following treatment with orthophosphate (dose: 2.6 mg-PO4/L, pH: 7.9), pH adjustment (pH: 7.9) and sodium silicate (dose: 20 mg-SiO2/L, pH: 7.9), respectively. When silica dose was increased from 20 mg-SiO2/L to 25 mg-SiO2/L (pH: 8.1), lead release destabilized and increased (median dissolved lead: 141 µg/L) due to formation of colloidal dispersions composed mainly of lead- and aluminum-rich phases as detected by field flow fractionation used with inductively coupled plasma mass spectrometry. Si was present in the scale at a maximum of 2.2 atomic % after 17 weeks of silica dosing at 20 mg- SiO2/L. Under the conditions tested, sodium silicate did not offer any benefits for reducing lead release from this LSL other than increasing pH. However, sodium silicate resulted in lower levels of biofilm accumulation on pipe walls, as measured by heterotrophic plate counts, when compared to orthophosphate.

The Salmonella enterica Plasmidome as a Reservoir of Antibiotic Resistance.

The emergence of multidrug-resistant bacterial strains worldwide has become a serious problem for public health over recent decades. The increase in antimicrobial resistance has been expanding via plasmids as mobile genetic elements encoding antimicrobial resistance (AMR) genes that are transferred vertically and horizontally. This study focuses on Salmonella enterica, one of the leading foodborne pathogens in industrialized countries. S. enterica is known to carry several plasmids involved not only in virulence but also in AMR. In the current paper, we present an integrated strategy to detect plasmid scaffolds in whole genome sequencing (WGS) assemblies. We developed a two-step procedure to predict plasmids based on i) the presence of essential elements for plasmid replication and mobility, as well as ii) sequence similarity to a reference plasmid. Next, to confirm the accuracy of the prediction in 1750 S. enterica short-read sequencing data, we combined Oxford Nanopore MinION long-read sequencing with Illumina MiSeq short-read sequencing in hybrid assemblies for 84 isolates to evaluate the proportion of plasmid that has been detected. At least one scaffold with an origin of replication (ORI) was predicted in 61.3% of the Salmonella isolates tested. The results indicated that IncFII and IncI1 ORIs were distributed in many S. enterica serotypes and were the most prevalent AMR genes carrier, whereas IncHI2A/IncHI2 and IncA/C2 were more serotype restricted but bore several AMR genes. Comparison between hybrid and short-read assemblies revealed that 81.1% of plasmids were found in the short-read sequencing using our pipeline. Through this process, we established that plasmids are prevalent in S. enterica and we also substantially expand the AMR genes in the resistome of this species.

Response of sulfate-reducing bacteria and supporting microbial community to persulfate exposure in a continuous flow system.

Coupling of chemical oxidation using persulfate with bioremediation has been proposed as a method to increase remedial efficacy at petroleum hydrocarbon contaminated sites. To support this integrated treatment approach, an understanding of persulfate impact on the indigenous microbial community is necessary for system design. As sulfate-reducing bacteria (SRB) are active in most aquifer systems and can utilize the sulfate generated from the degradation of persulfate, this study assessed the impact on SRB and the supporting anaerobic microbial community when exposed to persulfate in a continuous flow system. A series of bioreactors (1000 L) packed with anaerobic aquifer material were operated for an 8 month acclimatization period before being continuously subjected to benzene, toluene, ethylbenzene and xylenes (total BTEX 3 mg L-1). After 2 months, the bioreactors were then exposed to an unactivated persulfate solution (20 g L-1), or an alkaline-activated persulfate solution (20 g L-1, pH 12) then effluent-sampled for 60 days following. A combination of culture and molecular-based techniques were used to monitor SRB presence and structural profiles in the anaerobic SRB-specific and broader microbial community. Post-exposure, the rate of BTEX mass removal remained below pre-exposure values; however, trends suggest that full recovery would be expected. Rebound of SRB-specific and the associated microbial community to pre-exposure levels were observed in all exposed bioreactors. Structural community profiles identified recovery in both microbial species and diversity indices. Findings from this investigation demonstrate robustness of SRB in the presence of a supporting microbial community and, thus, are suitable organisms for target use during bioremediation in an integrated system with persulfate.

Pathogen and indicator variability in a heavily impacted watershed.

Water samples were collected from 36 locations within the Grand River Watershed, in Southwestern Ontario, Canada from July 2002 to December 2003 and were analyzed for total coliforms, fecal coliforms, Escherichia coli, Escherichia coli O157:H7, and thermophilic Campylobacter spp. A subset of samples was also analyzed for Cryptosporidium spp., Giardia spp., culturable human enteric viruses, and Clostridium perfringens. Storm and snowmelt events were sampled at two locations including a drinking water intake. For the majority of the events, the Spearman rank correlation test showed a positive correlation between E. coli levels and turbidity. Peaks in pathogen numbers frequently preceded the peaks in numbers of indicator organisms and turbidity. Pathogen levels sometimes decreased to undetectable levels during an event. As pathogen peaks did not correspond to turbidity and indicator peaks, the correlations were weak. Weak correlations may be the result of differences in the sources of the pathogens, rather than differences in pathogen movement through the environment. Results from this investigation have implications for planning monitoring programs for water quality and for the development of pathogen fate and transport models to be used for source water risk assessment.

Effect of temperature and disinfection strategies on ammonia-oxidizing bacteria in a bench-scale drinking water distribution system.

The establishment of ammonia-oxidizing bacteria (AOB), a group of autotrophic microorganisms responsible for nitrification in chloraminated distribution systems, was studied in a bench-scale distribution system. The potential significance of temperature and disinfectant residual associated with chloramination in full-scale drinking water distribution systems was assessed. Biofilm development was primarily monitored using AOB abundance and nitrite concentrations. The bench-scale system was initially operated under typical North American summer (22 degrees C) and fall (12 degrees C) temperatures, representing optimal and less optimal growth ranges for these microorganisms. Additional experimentation investigated AOB establishment at a suboptimal winter distribution system temperature of 6 degrees C. The effect of chloramine residual on AOB establishment was studied at higher (0.2-0.6mg/L) and lower (0.05-0.1mg/L) ranges, using a 3:1 (w/w) chlorine:ammonia dosing ratio. Conditions were selected to represent those typically found in a North American distribution system, in areas of low flow and longer retention times, respectively. Finally, the effect of a free chlorine residual on an established nitrifying biofilm was briefly examined. Results clearly indicate that AOB development occurs at all examined temperatures, as well as at selected monochloramine residuals. The maintenance of a disinfectant residual was difficult at times, but was more inhibitory to the nitrifying biofilm than the lower temperature. It can be concluded from the data that nitrification may not be adequately inhibited during the winter months, which may result in more advanced stages of nitrification the following season. Free chlorination can be effective in controlling AOB activity in the short term, but may not prevent reestablishment of a nitrifying biofilm upon return to chloramination.

Effect of gold nanoparticles and ciprofloxacin on microbial catabolism: a community-based approach.

The effect of gold nanoparticles (AuNPs) and ciprofloxacin on the catabolism of microbial communities was assessed. This was accomplished through an ex situ methodology designed to give a priori knowledge on the potential for nanoparticles, or other emerging contaminants, to affect the catabolic capabilities of microbial communities in the environment. Microbial communities from a variety of sources were incubated with 31 prespecified carbon sources and either National Institute of Standards and Technology reference material 10-nm AuNPs or ciprofloxacin on 96-well microtiter plates. From the ciprofloxacin study, dose-response curves were generated and exemplified how this method can be used to assess the effect of a toxicant on overall catabolic capabilities of microbial communities. With 10-nm AuNPs at concentrations ranging from 0.01 µg/mL to 0.5 µg/mL, rhizosphere communities from Typha roots were only slightly catabolically inhibited at a single concentration (0.05 µg/mL); no effects were seen on wetland water communities, and a minor positive (i.e., enhanced catabolic capabilities) effect was observed for loamy soil communities. This positive effect might have been because of a thin layer of citrate found on these AuNPs that initiated cometabolism with some of the carbon sources studied. Under the conditions considered, the possible adverse effects of AuNPs on the catabolic capabilities of microbial communities appears to be minimal.

An evaluation of behavioral health compliance and microbial risk factors on student populations within a high-density campus.

OBJECTIVE: The aim of this Canadian study was to assess student behavioral response to disease transmission risk, while identifying high microbial deposition/transmission sites. PARTICIPANTS: A student survey was conducted during October 2009. METHODS: The methods included a survey of students to assess use of health services, vaccination compliance, and hygiene along with a microbial analysis of potential transmission sites targeting specific residence buildings on campus. RESULTS: Results indicated that most students maintained that they were worried about H1N1 and reported making changes in hygienic behavior, with the majority not planning to be vaccinated. The microbial analysis indicated contamination of fomites in co-ed residences to be higher than either male or female student residences. CONCLUSIONS: A consideration of physical space along with behavioral factors is required in order to properly assess risk pathways in the establishment of an evidence-based infection control plan for universities and their contiguous communities.

Effect of ciprofloxacin on microbiological development in wetland mesocosms.

An understanding of how antibiotics and other "emerging contaminants" affect both water treatment systems and natural environments is of growing interest. Ciprofloxacin is a broad-spectrum antibiotic active against both Gram-positive and Gram-negative bacteria and has been extensively used over the past 20 years. The objective of this research was to study the effect of an antibiotic such as ciprofloxacin on the development, function and stability of bacterial communities in wetland systems. Four mesocosm wetlands planted with Phragmites australis were initially seeded with activated sludge from a waste water treatment plant and allowed to develop for a 1 week period, after which 2 of the 4 mesocosms were exposed to a ciprofloxacin concentration of 2 µg/mL for a 5 day period. The 4 mesocosms were then monitored for several microbiological and hydrological parameters over the course of 22 weeks. The bacterial community species distribution and catabolic capabilities were assessed via denaturing gradient gel electrophoresis (DGGE) and community level physiological profiling (CLPP), based on carbon utilization. DGGE results indicated that the ciprofloxacin decreased the total number of PCR-amplifiable DNA (bacteria) and the overall diversity of bacterial operational taxonomic units (OTUs). Through CLPP it was shown that the interstitial microbiological community was initially adversely affected by the ciprofloxacin, creating a temporary decrease in the activity and overall catabolic capabilities of the inherent wetland bacterial communities. However, after a 2-5 week recovery period the activities and catabolic capabilities of the bacterial communities exposed to ciprofloxacin returned to levels comparable to those found for bacterial communities not exposed to the ciprofloxacin. These findings suggest that ciprofloxacin exposure may have an adverse effect on the inherent bacterial communities in wetland systems initially reducing their ability to assimilate anthropogenic carbon-based compounds; however, normal functionality may resume after a 2-5 week period. It was also observed that plants in the ciprofloxacin-treated mesocosms did not adapt to the antibiotic presence, instead showing initial browning of above ground parts and eventual die-off. Reduced porosity, evapotranspiration, and overall hydrological mixing in the ciprofloxacin-treated mesocosms was observed.

Hydrologic modeling of pathogen fate and transport.

A watershed-scale fate and transport model has been developed for Escherichia coli and several waterborne pathogens: Cryptosporidiumspp., Giardiaspp., Campylobacter spp, and E. coli O157:H7. The objectives were to determine the primary sources of pathogenic contamination in a watershed used for drinking water supply and to gain a greater understanding of the factors that most influence their survival and transport. To predict the levels of indicator bacteria and pathogens in surface water, an existing hydrologic model, WATFLOOD, was augmented for pathogen transport and tested on a watershed in Southwestern Ontario, Canada. The pathogen model considered transport as a result of overland flow, subsurface flow to tile drainage systems, and in-stream routing. The model predicted that most microorganisms entering the stream from land-based sources enter the stream from tile drainage systems rather than overland transport. Although the model predicted overland transport to be rare, when it occurred, it corresponded to the highest observed and modeled microbial concentrations. Furthermore, rapid increases in measured E. coli concentrations during storm events suggested that the resuspension of microorganisms from stream sediments may be of equal or greater importance than land-based sources of pathogens.

A review of drinking-water-associated endotoxin, including potential routes of human exposure.

In the past decade efforts have been made to reduce the formation of harmful disinfection byproducts during the treatment and distribution of drinking water. This has been accomplished in part by the introduction of processes that involve the deliberate encouragement of indigenous biofilm growth in filters. In a controlled environment, such as a filter, these biofilms remove compounds that would otherwise be available as disinfection byproduct precursors or support uncontrolled biological activity in distribution systems. In the absence of exposure to chlorinated water, most biofilm bacteria are gram negative and have an outer layer that contains endotoxin. To date, outbreaks of waterborne endotoxin-related illness attributable to contamination of water used in hemodialysis procedures have been only infrequently documented, and occurrences linked to ingestion or through dermal abrasions could not be located. However, a less obvious conduit, that of inhalation, has been described in association with aerosolized water droplets. This review summarizes documented drinking-water-associated incidents of endotoxin exposure attributable to hemodialysis and inhalation. Typical endotoxin levels in water and conditions under which substantial quantities can enter drinking water distribution systems are identified. It would appear that endotoxin originating in tap water can be inhaled but at present there is insufficient information available to quantify potential health risks.

Estimating potential environmental loadings of Cryptosporidium spp. and Campylobacter spp. from livestock in the Grand River Watershed, Ontario, Canada.

Exposure to waterborne pathogens in recreational or drinking water is a serious public health concern. Thus, it is important to determine the sources of pathogens in a watershed and to quantify their environmental loadings. The natural variability of potentially pathogenic microorganisms in the environment from anthropogenic, natural, and livestock sources is large and has been difficult to quantify. A first step in characterizing the risk of nonpoint source contamination from pathogens of livestock origin is to determine the potential environmental loading based on animal prevalence and fecal shedding intensity. This study developed a probabilistic model for estimating the production of Cryptosporidium spp. and Campylobacter spp. from livestock sources within a watershed. Probability density functions representing daily pathogen production rates from livestock were simulated for the Grand River Watershed in southwestern Ontario. The prevalence of pathogenic microorganisms in animals was modeled as a mixture of beta-distributions with parameters drawn from published studies. Similarly, gamma-distributions were generated to describe animal pathogen shedding intensity. Results demonstrate that although cattle are responsible for the largest amount of manure produced, other domesticated farm animals contribute large numbers of the two pathogenic microorganisms studied. Daily pathogen production rates are highly sensitive to the parameters of the gamma-distributions, illustrating the need for reliable data on animal shedding intensity. The methodology may be used for identifying source terms for pathogen fate and transport modeling and for defining and targeting regions that are most vulnerable to water contamination from pathogenic sources.