Effects of low or medium-pressure UV irradiation on the release of intracellular microcystin

UV irradiation could be an alternative growth inhibition treatment against toxic Microcystis blooms in lakes. This study examined the effect of UV irradiation on the release of toxic intracellular microcystin. Conventional algicidal treatment (e.g. copper sulfate) was known to cause rapid release of intracellular microcystin and additional problems, but no studies have examined the case of UV treatment. A pure culture of Microcystis aeruginosa PCC 7806 was exposed to monochromatic low-pressure or polychromatic medium-pressure UV lamps. Irradiated pure culture suspension was subsequently incubated for 14 days under white light fluorescent lamps. During incubation, profiles of the number of cells, intracellular and extracellular microcystin concentration were determined. After the UV exposure, the number of cells gradually diminished; the net log cell reduction after 6 days of incubation was 1.6 log or 2.0 log for 600 mJ/cm2 of LP or MP UV irradiation, respectively. There were three findings for UV irradiation effects on the release of intracellular microcystin. First, because UV exposure can inhibit increases in the number of cells for about 6 days, it inhibited the increase of microcystin concentrations in water. Second, intracellular microcystin was gradually released into the surrounding water through a gradual reduction in the number of cells, preventing a rapid increase of microcystin concentration in water. Microcystin concentration in water was not significantly higher in UV-irradiated samples than non-UV-irradiated sample. Third, UV irradiation was able to degrade intracellular microcystin; it was reduced from 24.6 to 7.06 or 7.16 fg/cell by 600 mJ/cm2 of LP or MP UV irradiation, respectively. This contributed to reduce increases in microcystin concentrations in water. UV treatment can inhibit Microcystis growth and reduce intracellular microcystin content without rapid release of intracellular microcystin.     

Genotyping of single Cryptosporidium oocysts in sewage by semi-nested PCR and direct sequencing

This study describes an approach for genotyping individual Cryptosporidium oocysts obtained from sewage. We isolated single immunofluorescent assay (IFA)-stained Cryptosporidium oocysts from sewage concentrate using glass capillary pipettes and inverted epifluorescence microscopy. Each isolated Cryptosporidium oocyst was analyzed by semi-nested PCR for the 18S rRNA gene and direct sequencing of the PCR products. A total of 74 of 107 oocysts isolated from sewage were genotyped successfully. Of the 74 genotyped isolates, 51% (38 oocysts) were identified as C. parvum genotype 1, 4% (3 oocysts) of C. parvum VF383 human isolates, 20% (15 oocysts) of C. parvum genotype 2, 14% (10 oocysts) of C. meleagridis, 7% (5 oocysts) of C. sp. Pig 1, 3% (2 oocysts) of C. sp PG1-26 pig isolates and 1% (1 oocyst) of C. parvum CPM1 isolated from mouse. The results of this study demonstrate that 18S rRNA-based semi-nested PCR and direct sequencing can be used to characterize individual Cryptosporidium oocysts and also to reveal the distribution of Cryptosporidium genotypes in environmental waters.     

Changes of physical and biochemical properties of Cryptosporidium oocysts with various storage conditions

Physical and biochemical properties of Cryptosporidium parvum oocyst were examined after storage under various conditions. Oocyst-positive-fecal samples recovered from calves were either stored in a 2.0% potassium dichromate solution (Cr) or deionized water (W), or kept as a fecal pellet (P), and stored at 4 or 18 degrees C for a maximum of 100 days. When stored in Cr at 4 degrees C, the morphology of oocysts and their ability to withstand ultrasonics was not affected by the storing media or the storage period. However, when stored at 18 degees C as a fecal pellet, the specific gravity of the oocysts increased and a significant decrease in the oocysts resistance to ultrasonics occurred. These changes in oocyst properties may affect the performance of methods used to detect oocysts in water samples. When using the current test methods or when developing a new test method, it is important to take these factors into consideration.     

UV-induced dark repair mechanisms in bacteria associated with drinking water

Caulobacter crescentus and Aquabacterium commune, both isolated from drinking water, as well as environmental isolates of Pseudomonas aeruginosa and Enterococcus faecium were treated with different UV fluences to study their capacity to restore induced DNA damages. Here, the induction of a key mechanism of bacterial dark repair, the so-called recA system, was analysed. With newly designed probes, the specific recA mRNA was detected by Northern blot. Additionally, the RecA protein was measured by the Western blot technique using a specific antibody. In drinking water bacteria as well as in opportunistic microorganisms, a specific induction of dark repair mechanisms was found even at UV fluences higher than 400J/m(2), the German standard for UV disinfection. This induction depended on the incubation time after UV treatment. Nevertheless, the UV-induced recA expressions were found to differ in the bacteria under investigation.     

Removal of viable and inactivated Cryptosporidium by dual- and tri-media filtration

The limited efficacy of disinfectants, other than ultraviolet irradiation and ozonation, as a barrier against Cryptosporidium parvum in drinking water treatment has underscored the increased importance of oocyst removal by filtration. Currently, no reliable surrogates have been identified for C. parvum removal by filtration. As a result, evaluations of the Cryptosporidium removal by treatment operations have been performed using oocysts. It has typically been assumed that chemically inactivated oocysts are suitable surrogates for viable oocysts. Measurements of electrophoretic mobility, however, have shown that chemical inactivation changes the surface charge of Cryptosporidium oocysts. The present bench-scale research indicated that formalin-inactivated oocysts are reliable surrogates for viable oocysts during both stable filter operation and periods where filtration processes are challenged, such as coagulation failure. This finding is important because of the practical difficulties associated with using viable oocysts in filtration investigations. Poor coagulation conditions severely compromised removal of viable and inactivated oocysts by dual- and tri-media filters compared to stable operating conditions and filter ripening, emphasizing the importance of optimized chemical pre-treatment (coagulation) for the successful removal of oocysts during filtration. The treatment optimization experiments also indicated that tri-media filters offered only marginally higher oocyst removals than dual-media filters.     

Effects of light on sediment nutrient flux and water column nutrient stoichiometry in a shallow lake

The effects of light and temperature on nutrient cycling (silica (Si), nitrogen (N) and phosphorus (P)) between sediments and water in a shallow eutrophic lake (Loch Leven, Scotland), and consequent effects on water column nutrient stoichiometry, were assessed using a series of intact sediment core incubation experiments. Estimates of actual seasonal dark and light P-fluxes were assessed using 24-h incubations. Sediment-P uptake was observed in spring (7 degrees C) and release in autumn (12 degrees C) and summer (17 degrees C), with the highest release rates ( approximately 17 mgPO4-Pm(-2) sediment surface area d(-1)) occurring in summer. In a longer (21-day) experiment in which the effects of light (light (n=6) and dark (n=6)) and temperature (five 4-day cycles to represent: 7 degrees C-->13 degrees C-->23 degrees C-->13 degrees C-->7 degrees C) on water column nutrient concentrations were assessed, PO(4-)-P, total P (TP), SiO2 and total silica (TSi) concentrations in the water column were all significantly higher under dark conditions (ANOVA, alpha=0.05). NH4-N (ammonium N) water column concentrations were observed to be higher under dark conditions at low temperatures and higher under light conditions following a high-temperature (23 degrees C) treatment. No significant light effects were observed for water column total N (TN) concentration. Flux estimates for all nutrients measured are given. In terms of water column nutrient stoichiometry, TN:TP ratio was significantly higher under light conditions, TSi:TN was significantly lower under light conditions, and TSi:TP did not vary significantly between the dark and light treatments. The main processes acting to regulate diffusive nutrient release appeared to be photosynthetic elevation of bottom water pH and dissolved oxygen concentration (both significantly higher under light conditions) and direct microalgal sequestration. Thus, a feedback mechanism exists in recovering shallow lakes where benthic microalgae can affect the stoichiometry (to favour P/Si limitation) of the plankton, and also of the main source of nutrients back to the sediments via the disproportionate regulation of sediment P, Si and N release.     

Effects of low- or medium-pressure ultraviolet lamp irradiation on Microcystis aeruginosa and Anabaena variabilis

A pure culture of Microcystis aeruginosa or Anabaena variabilis, the representatives of water blooming algae, was exposed to low-pressure (LP) or medium-pressure (MP) UV lamps. Irradiated pure culture suspension was subsequently incubated for 7d under white light fluorescent lamps. During incubation, profiles of the number of cells, DNA damage and photosynthetic activity were determined. When UV fluence was 600mJ/cm(2), M. aeruginosa cell numbers decreased throughout the 7-d incubation period, to produce 1.5log reduction (LP) or 1.2log reduction (MP) compared with control. The amount of DNA damage was 2.02x10(-4) ESS/base (LP) and 3.42x10(-4) ESS/base (MP) just after UV irradiation, which became 0.05x10(-4) ESS/base and 0.23x10(-4) ESS/base, respectively, after 3d incubation. However, cell number kept decreasing, even after DNA repair. Photosynthetic activity decreased by 1.5log within 1d (LP) or 3d (MP). Thus, reduction in photosynthetic activity could contribute to the reduction in M. aeruginosa cell numbers. A. variabilis cell numbers reduced by 2.3log (LP) or 2.2log (MP) during the 7-d incubation period; however, after DNA damage repair, cell number began to increase. The amount of DNA damage was 6.07x10(-4) ESS/base (LP) and 4.48x10(-4) ESS/base (MP) just after UV irradiation, which became 0.23x10(-4) ESS/base and 0.40x10(-4) ESS/base, respectively, after 3d incubation. No reduction was observed in photosynthetic activity/cell. Therefore, DNA damage is the main contributor of the reduction in cell number of A. variabilis.     

The prevalence and characterisation of Cryptosporidium spp. in beef abattoir water supplies

The prevalence of Cryptosporidium spp. in 50 l samples of water used to wash beef carcasses at (a) an abattoir with a borehole water (BH) supply (n = 46) and (b) an abattoir with a river water (RW) supply (n = 48) was determined. In addition, a 100 l water sample and post-wash carcass samples (n = 24) were collected from the RW supply on a single day in July. Cryptosporidium spp. was detected in 0% and 26.1% of samples from the BH and RW supply abattoirs, respectively, with oocyst concentrations ranging from 0.02 to 8.6/l. Cryptosporidium spp. was not isolated from post-wash beef carcasses, while it was detected in water samples from that day at a concentration of 0.06 oocysts/l. The species of 3/5 isolates were identified as C. parvum, and the remaining were C. andersoni. This study has demonstrated that water used to wash beef carcasses can be contaminated with Cryptosporidium of human health importance and is a potential source of carcass contamination.     

Comparative disinfection efficiency of pulsed and continuous-wave UV irradiation technologies

Pulsed UV (PUV) is a novel UV irradiation system that is a non-mercury lamp-based alternative to currently used continuous-wave systems for water disinfection. PUV polychromatic irradiation disinfection efficiency was compared to that from continuous-wave monochromatic low-pressure (LP) and polychromatic medium-pressure (MP) UV systems, using two types of actinometry (ferrioxalate and iodide-iodate) and an absolute spectral emission method for fluence measurement. All three methods were in good agreement. Once accurate and reliable methods for fluence measurement were established, the inactivation of Escherichia coli and pathogen surrogates phage T4 and T7 were investigated under each technology. Inactivation was significantly faster using PUV irradiation compared to LP or MP UV lamps at equivalent fluence levels. A significant fraction of the enhanced PUV inactivation efficiency was due to wavelengths greater than 295 nm.     

MBR出水的紫外线消毒试验研究

考察了紫外线（UV）对MBR出水中微生物的灭活情况，以及光活化和暗修复对细菌灭活效果的影响。结果表明：UV对MBR出水中的微生物具有良好的灭活效果，当UV剂量为16mJ／cm^2时，对细菌的对数灭活率为3-lg；在相同的UV剂量下，不同UV强度对细菌的灭活效果无显著影响；经UV消毒后的细菌在3h内未出现明显的暗修复现象，但在日光灯和太阳光辐射下可发生明显的光活化现象，且不同光源下的光活化速率和达到饱和的时间有所不同。     

Laboratory tests of ammonium and barley straw extract as agents to suppress abundance of the harmful alga Prymnesium parvum and its toxicity to fish

Prymnesium parvum is a harmful alga whose blooms can cause fish kills in brackish waters. Two potential suppressants of this alga were tested, ammonium and barley straw extract (BSE), at temperatures of 10, 20 and 30 degrees C. Laboratory batch cultures were grown for 3 weeks at each temperature, with weekly doses of ammonium or BSE at either low or high levels, or a no-dose control treatment. The growth rate of P. parvum during exponential phase was highest at 20 degrees C and lowest at 10 degrees C, and was stimulated by the highest ammonium dose. Only cultures grown at 20 degrees C were toxic to fish. The highest ammonium dose abolished such toxicity and reduced the endpoint population density of P. parvum. BSE did not reduce the exponential growth rate, endpoint density, or toxicity to fish of P. parvum. The results support the use of ammonium additions, but not BSE, to suppress harmful blooms of P. parvum in those circumstances where the possible disadvantages can be managed.     

Settling velocity of Cryptosporidium parvum and Giardia lamblia

Understanding transport behavior of Cryptosporidium parvum oocysts and Giardia lamblia cysts (together referred to as (oo)cysts) in overland flow is important for beneficial uses of receiving water bodies. Like sediment, (oo)cysts are subjected to deposition once they are present in overland flow or low flow environments like reservoirs, wetlands and sedimentation basins. The objectives of this paper are to present the theory and experiment to determine the free settling velocity (v(s)) of (oo)cysts and to compare experimental settling velocities to estimates using Stokes' law. A settling experiment was designed to quantify the v(s) of (oo)cysts in an aqueous column. C. parvum oocysts used were spherical with average diameter (+/-1SD) of 6.6+/-1.1 microm. G. lamblia cysts were oval shaped (average eccentricity = 1.48+/-0.19) with average size of 11.8 +/-1.3 microm. Average densities were 1009 kg m(-3) for C. parvum oocysts and 1013 kg m(-3) for G. lamblia cysts. Observed experimental settling velocities are 0.27 microm s(-1) and 0.67 microm s(-1) for C. parvum and G. lamblia, respectively. Estimated average settling velocities using Stokes' law were 0.36 microm s(-1) for C. parvum and 0.84 microm s(-1) for G. lamblia. R-squared values of the observations from the settling experiments with the Stokes' law estimation are 0.87 and 0.88 for G. lamblia and C. parvum, respectively. Our results suggest that Stokes' law can be used to estimate settling velocities of (oo)cysts. Qualitatively, the low settling velocities indicate that (oo)cysts will very slowly settle out of suspension.     

Impact of bathers on levels of Cryptosporidium parvum oocysts and Giardia lamblia cysts in recreational beach waters

Recreational beach water samples collected on weekends and weekdays during 11 consecutive summer weeks were tested for potentially viable Cryptosporidium parvum oocysts and Giardia lamblia cysts using the multiplexed fluorescence in situ hybridization (FISH) method. The levels of oocysts and cysts on weekends were significantly higher than on the weekdays (P<0.01). Concentrations of oocysts in weekend samples (n=27) ranged from 2 to 42 oocysts/L (mean: 13.7 oocysts/L), and cyst concentration ranged from 0 to 33 cysts/L (mean: 9.1 cysts/L). For the samples collected on weekdays (n=33), the highest oocyst concentration was 7 oocysts/L (mean: 1.5 oocysts/L), and the highest cyst concentration was 4 cysts/L (mean: 0.6 cysts/L). The values of water turbidity were significantly higher on weekends than on weekdays, and were correlated with the number of bathers and concentration of C. parvum oocysts and G. lamblia cysts (P<0.04). The study demonstrated positive relationships between number of bathers and levels of waterborne C. parvum oocysts and G. lamblia cysts in recreational beach water. It is essential to test recreational waters for Cryptosporidium and Giardia when numbers of bathers are greatest, or limit the number of bathers in a recreational beach area.     

Capture of water-borne colloids in granular beds using external electric fields: improving removal of Cryptosporidium parvum

Suboptimal coagulation in water treatment plants often results in reduced removal efficiency of Cryptosporidium parvum oocysts by several orders of magnitude (J. AWWA 94(6) (2002) 97, J. AWWA 93(12) (2001) 64). The effect of external electric field on removal of C. parvum oocysts in packed granular beds was studied experimentally. A cylindrical configuration of electrodes, with granular media in the annular space was used. A negative DC potential was applied to the central electrode. No coagulants or flocculants were used and filtration was performed with and without application of an electric field to obtain improvement in removal efficiency. Results indicate that removal of C. parvum increased from 10% to 70% due to application of field in fine sand media and from 30% to 96% in MAGCHEM media. All other test particles (Kaolin and polystyrene latex microspheres) used in the study also exhibited increased removal in the presence of an electric field. Single collector efficiencies were also computed using approximate trajectory analysis, modified to account for the applied external electric field. The results of these calculations were used to qualitatively explain the trends in the experimental observations.     

Modelling of reactivation after UV disinfection: effect of UV-C dose on subsequent photoreactivation and dark repair

The increased use of UV radiation as a wastewater treatment technology has stimulated studies of the repair potential of microorganisms following treatment. In this study, samples of unfiltered secondary effluent were irradiated with seven levels of UV-C doses (50-200 mW s/cm(2)) from six low-pressure lamps in an open-channel UV disinfection system. Following irradiation, samples were incubated at 20 degrees C under photoreactivating light or in darkness. Samples were analysed for 240 min following incubation. The logistic model is proposed to explain the relation between photoreactivation and the UV-C dose received by the microorganisms. That model accurately fitted the data obtained in photoreactivation experiments, permitting interpretation of the estimated kinetic parameters: S(m) and k(2). In the experiments carried out in darkness, a slight reactivation is observed (<0.1%), followed by a decay period in which survival decreases. In order to model this last period, a modification was made to the logistic model by including a term of mortality that assumes a zero-order kinetic. The parameters S(m) and k(2), in both photoreactivation and darkness, show an exponential dependence on the UV-C inactivating dose. It is possible to predict their values, and hence the reactivation curve, from the equations proposed in this work.     

Biology, persistence and detection of Cryptosporidium parvum and Cryptosporidium hominis oocyst

Cryptosporidium parvum and Cryptosporidium hominis are obligate enteric protozoan parasites which infect the gastrointestinal tract of animals and humans. The mechanism(s) by which these parasites cause gastrointestinal distress in their hosts is not well understood. The risk of waterborne transmission of Cryptosporidium is a serious global issue in drinking water safety. Oocysts from these organisms are extremely robust, prevalent in source water supplies and capable of surviving in the environment for extended periods of time. Resistance to conventional water treatment by chlorination, lack of correlation with biological indicator microorganisms and the absence of adequate methods to detect the presence of infectious oocysts necessitates the development of consistent and effective means of parasite removal from the water supply. Additional research into improving water treatment and sewage treatment practices is needed, particularly in testing the efficiency of ozone in oocyst inactivation. Timely and efficient detection of infectious C. parvum and C. hominis oocysts in environmental samples requires the development of rapid and sensitive techniques for the concentration, purification and detection of these parasites. A major factor confounding proper detection remains the inability to adequately and efficiently concentrate oocysts from environmental samples, while limiting the presence of extraneous materials. Molecular-based techniques are the most promising methods for the sensitive and accurate detection of C. parvum and C. hominis. With the availability of numerous target sequences, RT-PCR will likely emerge as an important method to assess oocyst viability. In addition, a multiplex PCR for the simultaneous detection of C. parvum, C. hominis and other waterborne pathogens such as Giardia lamblia would greatly benefit the water industry and protect human health.     

Synergistic effect of sequential or combined use of ozone and UV radiation for the disinfection of Bacillus subtilis spores

This study was performed to evaluate the inactivation efficiency or synergy of combined ozone and UV processes (combined ozone/UV process) or sequential processes (ozone-UV, UV-ozone) compared with individual unit processes and to investigate the specific roles of ozone, UV and the hydroxyl radical, which is formed as an intermediate in the combined ozone/UV process. The Bacillus subtilis spore, which has often been used as a surrogate microorganism for Cryptosporidium parvum oocysts, was used as a target microorganism. Compared to individual unit processes with ozone or UV, the inactivation of B. subtilis spores by the combined ozone/UV process was enhanced under identical conditions. To investigate the specific roles of ozone and UV in the combined ozone/UV process, sequential ozone-UV and UV-ozone processes were tested for degrees of inactivation. Additionally, the experiment was performed in the presence and absence of tert-butyl alcohol, which acted as a hydroxyl radical scavenger to assess the role of inactivation by the hydroxyl radical in the combined ozone/UV process. Among the five candidate processes, the greatest synergistic effect was observed in the combined ozone/UV process. From the comparison of five candidate processes, the hydroxyl radical and ozone were each determined to significantly enhance the overall inactivation efficiency in the combined ozone/UV process.     

Standardizing photoreactivation: comparison of DNA photorepair rate in Escherichia coli using four different fluorescent lamps

The disinfection of pathogens in water via ultraviolet light exposure is a growing treatment technology, although the potential repair of UV damage following disinfection is a continued topic of concern in many UV disinfection applications. The conditions for investigating DNA repair of microorganisms are not standardized nor often correctly reported in the literature resulting in contradicting results regarding repair abilities of specific microorganisms. In this work a protocol was developed that enables comparison of photorepair experiments from different laboratories. Four different commonly used lamp types as well as sunlight were investigated to induce photorepair in Escherichia coli. Both time-based and photorepair fluence-based comparisons were performed. It was concluded that both intensity and lamp spectra significantly affect photorepair rate. Based on photorepair fluence calculations, there was no significant difference in photorepair rate between the different lamp types or compared to sunlight, while large differences were indicated using a time-based comparison as typically reported in the literature. Interestingly, the lethal effects of solar light on E. coli overshadowed the photorepair, and after 10 min the number of E. coli cells started to decrease. Because the disinfection efficiency of the sun affected the results, laboratory-based repair experiments may only indicate "potential" photorepair, as sunlight may be more lethal than laboratory photorepair lamps.     

Modeling Cryptosporidium parvum oocyst inactivation and bromate in a flow-through ozone contactor treating natural water

A reactive transport model was developed to simultaneously predict Cryptosporidium parvum oocyst inactivation and bromate formation during ozonation of natural water. A mechanistic model previously established to predict bromate formation in organic-free synthetic waters was coupled with an empirical ozone decay model and a one-dimensional axial dispersion reactor (ADR) model to represent the performance of a lab-scale flow-through ozone bubble-diffuser contactor. Dissolved ozone concentration, bromate concentration (in flow-through experiments only), hydroxyl radical exposure and C. parvum oocyst survival were measured in batch and flow-through experiments performed with filtered Ohio River water. The model successfully represented ozone concentration and C. parvum oocyst survival ratio in the flow-through reactor using parameters independently determined from batch and semi-batch experiments. Discrepancies between model prediction and experimental data for hydroxyl radical concentration and bromate formation were attributed to unaccounted for reactions, particularly those involving natural organic matter, hydrogen peroxide and carbonate radicals. Model simulations including some of these reactions resulted in closer agreement between predictions and experimental observations for bromate formation.