Coupled factors influencing the transport and retention of Cryptosporidium parvum oocysts in saturated porous media

The coupled role of solution ionic strength (IS), hydrodynamic force, and pore structure on the transport and retention of viable Cryptosporidium parvum oocyst was investigated via batch, packed-bed column, and micromodel systems. The experiments were conducted over a wide range of IS (0.1-100 mM), at two Darcy velocities (0.2 and 0.5 cm/min), and in two sands (median diameters of 275 and 710 μm). Overall, the results suggested that oocyst retention was a complex process that was very sensitive to the solution IS, the Darcy velocity, and the grain size. Increasing IS led to enhanced retention of oocysts in the column, which is qualitatively consistent with predictions of Derjaguin-Landau-Verwey-Overbeek theory. Conversely, increasing velocity and grain size resulted in less retention of oocysts in the column due to the difference in the fluid drag force and the rates of mass transfer from the liquid to the solid phase and from high to low velocity regions. Oocyst retention was controlled by a combined role of low velocity regions, weak attractive interactions, and/or steric repulsion. The contribution of each mechanism highly depended on the solution IS. In particular, micromodel observations indicated that enhanced oocyst retention occurred in low velocity regions near grain-grain contacts under highly unfavorable conditions (IS = 0.1 mM). Oocyst retention was also found to be influenced by weak attractive interactions (induced by the secondary energy minimum, surface roughness, and/or nanoscale chemical heterogeneity) when the IS = 1 mM. Reversible retention of oocysts to the sand in batch and column studies under favorable attachment conditions (IS = 100 mM) was attributed to steric repulsion between the oocysts and the sand surface due to the presence of oocyst surface macromolecules. Comparison of experimental observations and theoretical predictions from classic filtration theory further supported the presence of this weak interaction due to steric repulsion.     

Transport of selected bacterial pathogens in agricultural soil and quartz sand

The protection of groundwater supplies from microbial contamination necessitates a solid understanding of the key factors controlling the migration and retention of pathogenic organisms through the subsurface environment. The transport behavior of five waterborne pathogens was examined using laboratory-scale columns packed with clean quartz at two solution ionic strengths (10 mM and 30 mM). Escherichia coli O157:H7 and Yersinia enterocolitica were selected as representative Gram-negative pathogens, Enterococcus faecalis was selected as a representative Gram-positive organism, and two cyanobacteria (Microcystis aeruginosa and Anabaena flos-aquae) were also studied. The five organisms exhibit differing attachment efficiencies to the quartz sand. The surface (zeta) potential of the microorganisms was characterized over a broad range of pH values (2-8) at two ionic strengths (10 mM and 30 mM). These measurements are used to evaluate the observed attachment behavior within the context of the DLVO theory of colloidal stability. To better understand the possible link between bacterial transport in model quartz sand systems and natural soil matrices, additional experiments were conducted with two of the selected organisms using columns packed with loamy sand obtained from an agricultural field. This investigation highlights the need for further characterization of waterborne pathogen surface properties and transport behavior over a broader range of environmentally relevant conditions.     

The effects of biofilm on the transport of stabilized zerovalent iron nanoparticles in saturated porous media

The transport of stabilized zerovalent iron nanoparticles (nZVI) has recently been the topic of extensive research due to its proven potential as an in situ remediation tool. However, these studies have ignored the effects of biofilms—complex aggregations of bacterial cells and excreted extracellular polymeric substances present in nearly all aquatic systems—on the transport of these particles. This study examines the effects of Pseudomonas aeruginosa (PAO1) biofilm, at a cell concentration similar to that reported for saturated aquifers, on the transport of commercially available, poly (acrylic acid) stabilized nZVI (pnZVI) in 14 cm long columns packed with saturated glass beads at salt concentrations of 1 and 25 mM NaCl. Compared to retention on uncoated columns, in the presence of biofilm the retention of pnZVI increased at higher ionic strength, while ionic strength played no role in retention of these nanoparticles in the absence of biofilm. The Tufenkji-Elimelech correlation equation predicts lower retention of pnZVI on biofilm coated columns compared to uncoated columns due to a lower Hamaker constant, and DLVO energy considerations predict the most favorable attachment to uncoated porous media at higher ionic strength. A steric (polymer-mediated) model that considers the combined influence of steric effects of polymers and DLVO interactions is shown to adequately describe particle retention in columns.     

Towards understanding inter-strain attachment variations of Escherichia coli during transport in saturated quartz sand

Although Escherichia coli is an indicator of fecal contamination in aquifers, limited research has been devoted to understanding the biological processes involved in the initial attachment of E. coli transported in abiotic porous media. The roles of the various surface structures of E. coli, like lipopolysaccharides (LPS), autotransporter proteins, and fimbriae are unknown. The objective of this research was to establish the effects of variations in surface characteristics of the outer membrane of E. coli on the attachment efficiency of 54 E. coli strains upon transport in saturated quartz sand under identical flow conditions. We used column experiments to assess retention of the E. coli strains, and we determined sphericity, motility, zeta-potential, and aggregation of all strains. LPS composition was determined based on known serotypes, and the presence/absence of 22 genes encoding surface characteristics was determined with qualitative PCR. The results indicated that under identical flow conditions, there was a variation of two orders of magnitude in the maximum breakthrough concentrations of the 54 E. coli strains. Of all factors we investigated, no single factor was able to explain attachment efficiency variations statistically significantly. However, low attachment efficiencies were associated (p = 0.13) with LPS containing saccharides with phosphate and/or carboxyl groups. These saccharide groups are acidic and likely charged with a negative O-atom, which reduced attachment to the negatively charged quartz surface. In addition, of the 22 genes tested, Afa was most associated (p = 0.21) with attachment efficiency. The work presented here bridges knowledge on colloid transport and molecular microbiology, and tries to offer a more holistic view on the attachment of planktonic E. coli bacteria to (abiotic) quartz grain surfaces. Future research should involve the use of microbiological techniques in order to be able to map the unique or grouped characteristics of E. coli in aquifers, and to assess the usefulness of E. coli as a fecal indicator in aquifers.     

Production of Shiga-like toxins in viable but nonculturable Escherichia coli O157:H7

Escherichia coli O157:H7, a causative agent of hemolytic uremic syndrome, can enter into a viable but nonculturable (VBNC) state when under stress. To date, it is unknown whether VBNC cells produce Shiga-like toxins (Stx). To address this question, we confirmed the expression of the stx1 and stx2 genes and the production of Stx in VBNC E. coli O157:H7 cells. To quantitatively assess the production of Stx in VBNC cells, we developed a Vero-cell microplate cytotoxicity assay based on the correspondence of the cytotoxicity of VBNC cells on Vero cells to the number of inoculated VBNC cells. Using this method, we found that all VBNC cells induced by river water, PBS buffer, deionized water, or chloraminated water retained the ability to produce Stx, and that they had differing levels of Stx. Both aged (19-month-old) VBNC cells induced by river water and fresh VBNC cells induced by chloraminated water showed very low half maximal inhibitory concentration (IC₅₀; 6.6 × 10⁴ and 7.1 × 10⁴ respectively), corresponding to higher levels of toxins produced than VBNC cells induced by deionized water and PBS buffer. VBNC cells originating from different isolates may vary in Stx production, and the VBNC cells from bovine isolates produced higher levels of Stx than those from clinical isolates. These results demonstrate a potential health risk of VBNC E. coli O157:H7 in environmental water and the importance of monitoring VBNC E. coli O157:H7.     

Effects of fumigants on microbial diversity and persistence of E. coli O15:H7 in contrasting soil microcosms

Persistence of E. coli O157 in the environment is a serious public health concern. However, little is known about the persistence of this pathogen after exposure to chemical compounds like fumigants in the environment. In this study, the persistence behavior of pathogenic E. coli O157:H7 was investigated after fumigation with methyl bromide (MeBr; CH₃Br) and methyl iodide (MeI, iodomethane; CH₃I) in soil microcosms under laboratory conditions. Our goal was to assess changes in soil microbial community structure and persistence of E. coli O157:H7 in microcosm soils after fumigation. PCR was used to amplify 16S rRNA genes from total bacterial community composition, and the products were subjected to denaturing gradient gel electrophoresis (DGGE). Microbial diversity as determined by DGGE was significantly higher in clay soil than sandy soil. Real-time PCR and plate counts were used to quantify the survival of E. coli O157:H7 in the two soils after fumigation with MeBr and MeI. The survival of the pathogen was higher in the non fumigated controls than the fumigated treatments when determined using plate counts. These results were confirmed by real time PCR analysis targeting the stx1, stx2, and the eae genes. E. coli O157:H7 survived for about 35 days when determined using the plate count method but continued to be detected at about the detection limit of 10² by real time PCR for more than 86 days. Our results showed that there was a fast inactivation of the pathogen during the first 35 days. After this period, a small proportion of the pathogen continued to survive in the soil microcosms. Subsequent enrichment of soil samples and immunomagnetic separation revealed the continuous presence of viable cells after 86 days of incubation. The data presented contribute to a better understanding of the behavior of E. coli O157:H7 in soil, and showed the need for more investigation of the role of dormant cells in soil that may be a source for recontamination of the environment.     

Transport of two metal oxide nanoparticles in saturated granular porous media: role of water chemistry and particle coating

The growing use of nanosized titanium dioxide (nTiO₂) and zinc oxide (nZnO) in a large number of commercial products raises concerns regarding their release and subsequent mobility in natural aquatic environments. Laboratory-scale sand-packed column experiments were conducted with bare and polymer-coated nTiO₂ and nZnO to improve our understanding of the mobility of these nanoparticles in natural or engineered water saturated granular systems. The nanoparticles are characterized over a range of environmentally relevant water chemistries using multiple complimentary techniques: dynamic light scattering, nanoparticle tracking analysis, transmission electron microscopy, and scanning electron microscopy. Overall, bare (uncoated) nanoparticles exhibit high retention within the water saturated granular matrix at solution ionic strengths (IS) as low as 0.1 mM NaNO₃ for bare nTiO₂ and 0.01 mM NaNO₃ for bare nZnO. Bare nTiO₂ and nZnO also display dynamic (time-dependent) deposition behaviors under selected conditions. In contrast, the polymer-coated nanoparticles are much less likely to aggregate and exhibit significant transport potential at IS as high as 100 mM NaNO₃ or 3 mM CaCl₂. These findings illustrate the importance of considering the extent and type of surface modification when evaluating metal oxide contamination potential in granular aquatic environments.     

Influence of tetracycline resistance on the transport of manure-derived Escherichia coli in saturated porous media

In this research, tetracycline resistant (tet^R) and tetracycline susceptible (tet^S) Escherichia coli isolates were retrieved from dairy manure and the influence of tetracycline resistance on the transport of E. coli in saturated porous media was investigated through laboratory column transport experiments. Experimental results showed that tet^RE. coli strains had higher mobility than the tet^S strains in saturated porous media. Measurements of cell surface properties suggested that tet^RE. coli strains exhibited lower zeta potentials than the tet^S strains. Because the surface of clean quartz sands is negatively charged, the repulsive electrostatic double layer (EDL) interaction between the tet^R cells and the surface of sands was stronger and thus facilitated the transport of the tet^R cells. Although no difference was observed in surface acidity, cell size, lipopolysaccharides (LPS) sugar content and cell-bound protein levels between the tet^R and tet^S strains, they displayed distinct outer membrane protein (OMP) profiles. It was likely that the difference in OMPs, some potentially related to drug efflux pumps, between the tet^R and tet^S strains led to alteration in cell surface properties which in turn affected cell transport in saturated porous media. Findings from this research suggested that manure-derived tet^RE. coli could spread more widely in the groundwater system and pose serious public health risks.     

On the role of metabolic activity on the transport und deposition of Pseudomonas fluorescens in saturated porous media

A study was conducted to understand the role of cell concentration and metabolic state in the transport and deposition behaviour of Pseudomonas fluorescens with and without substrate addition. Column experiments using the short-pulse technique (pulse was equivalent to 0.028 pore volume) were performed in quartz sand operating under saturated conditions. For comparison, experiments with microspheres and inactive (killed) bacteria were also conducted. The effluent concentrations, the retained particle concentrations and the cell shape were determined by fluorescent microscopy. For the transport of metabolically-active P. fluorescens without substrate addition a bimodal breakthrough curve was observed, which could be explained by the different breakthrough behaviour of the rod-shaped and coccoidal cells of P. fluorescens. The 70:30 rod/coccoid ratio in the influent drastically changed during the transport and it was about 20:80 in the effluent and in the quartz sand packing. It was assumed that the active rod-shaped cells were subjected to shrinkage into coccoidal cells. The change from active rod-shaped cells to coccoidal cells could be explained by oxygen deficiency which occurs in column experiments under saturated conditions. Also the substrate addition led to two consecutive breakthrough peaks and to more bacteria being retained in the column. In general, the presence of substrate made the assumed stress effects more pronounced. In comparison to microspheres and inactive (killed) bacteria, the transport of metabolically-active bacteria with and without substrate addition is affected by differences in physiological state between rod-shaped and the formed stress-resistant coccoidal cells of P. fluorescens.     

Impact of urbanization and agriculture on the occurrence of bacterial pathogens and stx genes in coastal waterbodies of central California

Fecal pollution enters coastal waters through multiple routes, many of which originate from land-based activities. Runoff from pervious and impervious land surfaces transports pollutants from land to sea and can cause impairment of coastal ocean waters. To understand how land use practices and water characteristics influence concentrations of fecal indicator bacteria (FIB) and pathogens in natural waters, fourteen coastal streams, rivers, and tidal lagoons, surrounded by variable land use and animal densities, were sampled every six weeks over two years (2008 & 2009). Fecal indicator bacteria (FIB; Escherichia coli and Enterococci) and Salmonella concentrations, the occurrence of Bacteroidales human, ruminant, and pig-specific fecal markers, E. coli O157:H7, and Shiga toxin (stx) genes present in E. coli, were measured. In addition, environmental and climatic variables (e.g., temperature, salinity, rainfall), as well as human and livestock population densities and land cover were quantified. Concentrations of FIB and Salmonella were correlated with each other, but the occurrence of host-specific Bacteroidales markers did not correlate with FIB or pathogens. FIB and Salmonella concentrations, as well as the occurrence of E. coli harboring stx genes, were positively associated with the fraction of the surrounding subwatershed that was urban, while the occurrence of E. coli O157:H7 was positively associated with the agricultural fraction. FIB and Salmonella concentrations were negatively correlated to salinity and temperature, and positively correlated to rainfall. Areal loading rates of FIB, Salmonella and E. coli O157:H7 to the coastal ocean were calculated for stream and river sites and varied with land cover, salinity, temperature, and rainfall. Results suggest that FIB and pathogen concentrations are influenced, in part, by their flux from the land, which is exacerbated during rainfall; once waterborne, bacterial persistence is affected by water temperature and salinity.     

Role of morphology in the aggregation kinetics of ZnO nanoparticles

The aggregation kinetics of two types of ZnO nanoparticles were investigated under various conditions. Distinct differences in aggregation kinetics were observed between the two ZnO particles. The aggregation of the nearly spherical ZnO (denoted as Me ZnO) exhibited strong dependence on the ionic strength (IS) of the solution; while minimal influence of IS was seen on the irregularly shaped ZnO (mixture of slab-like and rod-shaped particles, denoted as Mk ZnO) in the IS ranged tested. It is postulated that Mk ZnO possesses a critical coagulation concentration (CCC) below the lowest electrolyte concentration tested (1 mM NaCl) due to the interactions between various surfaces. The CCC of ZnO was found to be a function of pH; the CCC increased significantly as the pH was further away from the point of zero charge. Natural organic matter (NOM) was found to substantially hinder the aggregation of both types of ZnO particles (above 10 mg/L for Me ZnO and above 1 mg/L for Mk ZnO). A Langmuir adsorption model was used to describe the NOM to ZnO nanoparticle adsorption isotherms. To our knowledge, this is the first study to report the effect of particle morphology on nanoparticle aggregation, which outlines the importance of accounting morphology into environmental transport assessment of nanoparticles.     

Facilitated transport of Cu with hydroxyapatite nanoparticles in saturated sand: effects of solution ionic strength and composition

Column experiments were conducted to investigate the facilitated transport of Cu in association with hydroxyapatite nanoparticles (nHAP) in water-saturated quartz sand at different solution concentrations of NaCl (0-100 mM) or CaCl₂ (0.1-1.0 mM). The experimental breakthrough curves and retention profiles of nHAP were well described using a mathematical model that accounted for two kinetic retention sites. The retention coefficients for both sites increased with the ionic strength (IS) of a particular salt. However, the amount of nHAP retention was more sensitive to increases in the concentration of divalent Ca²⁺ than monovalent Na⁺. The effluent concentration of Cu that was associated with nHAP decreased significantly from 2.62 to 0.17 mg L⁻¹ when NaCl increased from 0 to 100 mM, and from 1.58 to 0.16 mg L⁻¹ when CaCl₂ increased from 0.1 to 1.0 mM. These trends were due to enhanced retention of nHAP with changes in IS and ionic composition (IC) due to compression of the double layer thickness and reduction of the magnitude of the zeta potentials. Results indicate that the IS and IC had a strong influence on the co-transport behavior of contaminants with nHAP nanoparticles.     

Survival of Escherichia coli in two sewage treatment plants using UV irradiation and chlorination for disinfection

We investigated the survival of Escherichia coli in two STPs utilising UV irradiation (STP-A) or chlorination (STP-B) for disinfection. In all, 370 E. coli strains isolated from raw influent sewage (IS), secondary treated effluent (STE) and effluent after the disinfection processes of both STPs were typed using a high resolution biochemical fingerprinting method and were grouped into common (C-) and single (S-) biochemical phenotypes (BPTs). In STP-A, 83 BPTs comprising 123 isolates were found in IS and STE, of which 7 BPTs survived UV irradiation. Isolates tested from the same sites of STP-B (n = 220) comprised 122 BPTs, however, only two BPTs were found post-chlorination. A representative isolate from each BPT from both STPs was tested for the presence of 11 virulence genes (VGs) associated with uropathogenic (UPEC) or intestinal pathogenic (IPEC) E. coli strains. Strains surviving UV irradiation were distributed among seven phylogenetic groups with five BPTs carrying VGs associated with either UPEC (4 BPTs) or IPEC (1 BPT). In contrast, E. coli strains found in STP-B carried no VGs. Strains from both STPs were resistant to up to 12 out of the 21 antibiotics tested but there was no significant difference between the numbers of antibiotics to which surviving strains were resistant to in these STPs. Our data suggests that some E. coli strains have a better ability to survive STPs utilising chlorination and UV irradiation for disinfection. However, strains that survive UV irradiation are more diverse and may carry more VGs than those surviving SPTs using chlorination.     

Molecular subtypes of Campylobacter spp., Salmonella enterica, and Escherichia coli O157:H7 isolated from faecal and surface water samples in the Oldman River watershed, Alberta, Canada

Campylobacter spp., Salmonella enterica, and Escherichia coli O157:H7 isolated from 898 faecal, 43 sewage, and 342 surface water samples from the Oldman River were characterized using bacterial subtyping methods in order to investigate potential sources of contamination of the watershed. Among these pathogens, Campylobacter spp. were the most frequently isolated from faecal, sewage, and surface water samples (266/895, 11/43, and 91/342, respectively), followed by Salmonella (67/898, 8/43, and 29/342, respectively), and E. coli O157:H7 (16/898, 2/43, and 8/342, respectively). Salmonella Rubislaw was the most common serovar isolated from water. This serovar was also isolated from two wild bird species. Most other serovars isolated from water were either not isolated from animals or were isolated from multiple species. E. coli O157:H7 was predominantly isolated from cattle. The most common phage-types of this pathogen from cattle were also the most common among water isolates, and there were exact pulsed field gel electrophoresis and comparative genomic fingerprint matches between cattle, sewage, and water isolates. Campylobacters were commonly isolated from surface waters and faeces from most animal species. Restriction fragment length polymorphism of the Campylobacter flaA gene identified several location and host species-specific (cattle, goose, pig) fingerprints. Molecular subtyping of these bacterial pathogens shows considerable promise as a tool for determining the sources of faecal pollution of water.     

Effects of primary sludge particulate (PSP) entrapment on ultrasonic (20 kHz) disinfection of Escherichia coli

The role of primary sludge particulates (PSPs) in ultrasonic disinfection of Escherichia coli (E. coli) was investigated. Entrapment of E. coli by PSP was directly observed through scanning electron microscope (SEM) after E. coli and PSP were incubated together in water for 24 h at 35 °C. Entrapment coefficient was proposed for the first time to reflect the ability of PSP to entrap E. coli and was estimated as 1.4 × 10³ CFU/mg PSP under our experimental conditions. Ultrasonication (20 kHz) of different E. coli-PSPs solutions showed that the entrapped E. coli cells were protected by PSP from ultrasonication and the unentrapped cells were not. However, the protection of entrapped E. coli cells gradually decreased as ultrasonication proceeded, suggesting the ability of power ultrasonication to deprotect the entrapped E. coli cells. SEM studies suggested a two-step mechanism for ultrasonic (20 kHz) disinfection of entrapped E. coli: breakdown of the protective PSP refugia and disinfection of the exposed E. coli cells. This research will enable more informed decisions about disinfection of aqueous samples where porous PSP are present.     

Bacteria,viruses, and parasites in an intermittent stream protected from and exposed to pasturing cattle: Prevalence,densities, and quantitative microbial risk assessment

Over 3500 individual water samples, for 131 sampling times, targeting waterborne pathogens/fecal indicator bacteria were collected during a 7-year period from 4 sites along an intermittent stream running through a small livestock pasture system with and without cattle access-to-stream restriction measures. The study assessed the impact of cattle pasturing/riparian zone protection on: pathogen (bacterial, viral, parasite) occurrence, concentrations of fecal indicators, and quantitative microbial risk assessments (QMRA) of the risk of Cryptosporidium, Giardia and Escherichia coli O157:H7 infection in humans. Methodologies were developed to compute QMRA mean risks on the basis of water samples exhibiting potentially human infectious Cryptosporidium and E. coli based on genotyping Crytosporidium, and E. coli O157:H7 presence/absence information paired with enumerated E. coli. All Giardia spp. were considered infectious. No significant pasturing treatment effects were observed among pathogens, with the exception of Campylobacter spp. and E. coli O157:H7. Campylobacter spp. prevalence significantly decreased downstream through pasture treatments and E. coli O157:H7 was observed in a few instances in the middle of the unrestricted pasture. Densities of total coliform, fecal coliform, and E. coli reduced significantly downstream in the restricted pasture system, but not in the unrestricted system. Seasonal and flow conditions were associated with greater indicator bacteria densities, especially in the summer. Norovirus GII was detected at rates of 7–22% of samples for all monitoring sites, and rotavirus in 0–7% of samples for all monitoring sites; pasture treatment trends were not evident, however. Seasonal and stream flow variables (and their interactions) were relatively more important than pasture treatments for initially stratifying pathogen occurrence and higher fecal indicator bacteria densities. Significant positive associations among fecal indicator bacteria and Campylobacter spp. detection were observed. For QMRA, adjusting for the proportion of Cryptosporidium spp. detected that are infectious for humans reduces downstream risk estimates by roughly one order of magnitude. Using QMRA in this manner provides a more refined estimate of beneficial management practice effects on pathogen exposure risks to humans.     

Searching for Escherichia coli O157 in drinking and recreational waters in Northern Greece

Escherichia coli O157:H7 has been recognized in sporadic cases and outbreaks, as an important cause of bloody diarrhea and also a leading cause of acute renal failure in children. The purpose of this study was to investigate the presence of E. coli O157 in drinking and recreational water samples from Northern Greece. For this, a total of 1974 colonies of E. coli that were isolated from 1267 water samples were recultured on sorbitol-MacConkey agar. Out of them 121 were found non-sorbitol fermenting, but none had the somatic O157 antigen. This finding supports the hypothesis that drinking and recreational waters should not be considered possible routes of E. coli O157:H7 transmission in Northern Greece.     

UV inactivation and characteristics after photoreactivation of Escherichia coli with plasmid: health safety concern about UV disinfection

Occurrence and degree of photoreactivation after ultraviolet (UV) exposure have been widely studied. However, the characteristics of photoreactivated microorganisms were rarely investigated. Hence, in this study, Escherichia coli with plasmids of ampicillin (amp)-resistance or fluorescence was used as indicators to examine the UV inactivation efficiencies and variations of characteristics of E. coli after subsequent photoreactivation.The experimental results indicate that the amp-resistant bacteria and the fluorescent bacteria used in this study had similar trends of UV dose-response curves. 3.5-log₁₀ and 3-log₁₀ reductions were achieved with a UV dose of 5 mJ/cm² for the amp-resistant and fluorescent E. coli, respectively. There was no significant difference in the UV inactivation behavior, as compared with common strains of E. coli.For the amp-resistant E. coli and the fluorescent E. coli, after exposures with UV doses of 5, 15, 25, 40 and 80 mJ/cm², the corresponding percent photoreactivations after a 4 h exposure to photoreactivating light were 1% and 46% respectively for a UV dose of 5 mJ/cm², and essentially negligible for all other UV doses. Furthermore, the photoreactivated amp-resistant bacteria still have the ability of amp-resistance. And the revived fluorescent E. coli showed similar fluorescent behavior, compared with the untreated bacteria. The experimental results imply that after UV inactivation and subsequent photoreactivation, the bacteria retained the initial characteristics coded in the plasmid. This reveals a possibility that some characteristics of bacteria can retain or recover through photoreactivation, and a safety concern about pathogenicity revival might need to be considered with UV disinfection and photoreactivation.     

Coupling of physical and chemical mechanisms of colloid straining in saturated porous media

Filtration theory does not include the potential influence of pore structure on colloid removal by straining. Conversely, previous research on straining has not considered the possible influence of chemical interactions. Experimental and theoretical studies were therefore undertaken to explore the coupling of physical and chemical mechanisms of colloid straining under unfavorable attachment conditions (pH=10). Negatively charged latex microspheres (1.1 and 3mum) and quartz sands (360, 240, and 150mum) were used in packed column studies that encompassed a range in suspension ionic strengths (6-106mM) and Darcy water velocities (0.1-0.45cmmin(-1)). Derjaguin-Landau-Verwey-Overbeek (DLVO) calculations and torque analysis suggests that attachment of colloids to the solid-water interface was not a significant mechanism of deposition for the selected experimental conditions. Effluent concentration curves and hyperexponential deposition profiles were strongly dependent on the solution chemistry, the system hydrodynamics, and the colloid and collector grain size, with greater deposition occurring for increasing ionic strength, lower flow rates, and larger ratios of the colloid to the median grain diameter. Increasing the solution ionic strength is believed to increase the force and number of colloids in the secondary minimum of the DLVO interaction energy profile. These weakly associated colloids can be funneled to small regions of the pore space formed adjacent to grain-grain junctions. For select systems, the ionic strength of the eluant solution was decreased to 6mM following the recovery of the effluent concentration curve. In this case, only a small portion of the deposited colloids was recovered in the effluent and the majority was still retained in the sand. These observations suggest that the extent of colloid removal by straining is strongly coupled to solution chemistry.     

Associations among pathogenic bacteria, parasites, and environmental and land use factors in multiple mixed-use watersheds

Over a five year period (2004-08), 1171 surface water samples were collected from up to 24 sampling locations representing a wide range of stream orders, in a river basin in eastern Ontario, Canada. Water was analyzed for Cryptosporidium oocysts and Giardia cyst densities, the presence of Salmonella enterica subspecies enterica, Campylobacter spp., Listeria monocytogenes, and Escherichia coli O157:H7. The study objective was to explore associations among pathogen densities/occurrence and objectively defined land use, weather, hydrologic, and water quality variables using CART (Classification and Regression Tree) and binary logistical regression techniques. E. coli O157:H7 detections were infrequent, but detections were related to upstream livestock pasture density; 20% of the detections were located where cattle have access to the watercourses. The ratio of detections:non-detections for Campylobacter spp. was relatively higher (>1) when mean air temperatures were 6% below mean study period temperature values (relatively cooler periods). Cooler water temperatures, which can promote bacteria survival and represent times when land applications of manure typically occur (spring and fall), may have promoted increased frequency of Campylobacter spp. Fifty-nine percent of all Salmonella spp. detections occurred when river discharge on a branch of the river system of Shreve stream order = 9550 was >83 percentile. Hydrological events that promote off farm/off field/in stream transport must manifest themselves in order for detection of Salmonella spp. to occur in surface water in this region. Fifty seven percent of L. monocytogenes detections occurred in spring, relative to other seasons. It was speculated that a combination of winter livestock housing, silage feeding during winter, and spring application of manure that accrued during winter, contributed to elevated occurrences of this pathogen in spring. Cryptosporidium and Giardia oocyst and cyst densities were, overall, positively associated with surface water discharge, and negatively associated with air/water temperature during spring-summer-fall. Yet, some of the highest Cryptosporidium oocyst densities were associated with low discharge conditions on smaller order streams, suggesting wildlife as a contributing fecal source. Fifty six percent of all detections of ≥2 bacteria pathogens (including Campylobacter spp., Salmonella spp., and E. coli O157:H7) in water was associated with lower water temperatures (<∼14 °C; primarily spring and fall) and when total rainfall the week prior to sampling was >∼27 mm (62 percentile). During higher water temperatures (>∼14 °C), a higher amount of weekly rainfall was necessary to promote detection of ≥2 pathogens (primarily summer; weekly rainfall ∼>42 mm (>77 percentile); 15% of all ≥2 detections). Less rainfall may have been necessary to mobilize pathogens from adjacent land, and/or in stream sediments, during cooler water conditions; as these are times when manures are applied to fields in the area, and soil water contents and water table depths are relatively higher. Season, stream order, turbidity, mean daily temperature, surface water discharge, cropland coverage, and nearest upstream distance to a barn and pasture were variables that were relatively strong and recurrent with regard to discriminating pathogen presence and absence, and parasite densities in surface water in the region.