Inactivation Of Cryptosporidium Parvum Oocysts Research Articles

Synergistic inactivation of Cryptosporidium parvum using ozone followed by free chlorine in natural water

The synergistic effect of sequential exposure to ozone followed by free chlorine on inactivation of Cryptosporidium parvum oocysts suspended in natural waters was studied in bench-scale batch reactors. Animal infectivity using neonatal CD-1 mice was used to measure oocyst inactivation. The synergistic effect measured in two alkaline (pH 8.1) natural waters was statistically significant but was considerably smaller than previously reported in buffered de-ionized water at pH 6.0. Temperature, ozone primary treatment level, and water type did not have measurable impacts on the synergistic effect. Efforts to increase the synergistic effect by reducing the pH from 8 to 6 by acid addition were unsuccessful. In the two low alkalinity (pH 6.0) natural waters tested, the measured synergistic effect was greater than in the alkaline waters, but was still less than that measured previously in buffered de-ionized water. It was concluded that the synergistic effect reduction in the natural waters tested was due in part to alkalinity and in part to other unidentified water quality characteristics. Sequential treatment with ozone followed by free chlorine may only be a feasible strategy for achieving synergistic C. parvum inactivation credit for water treatment facilities with natural waters having a low pH (near 6.0).

A Bayesian method of estimating kinetic parameters for the inactivation of Cryptosporidium parvum oocysts with chlorine dioxide and ozone

The main objective of this paper is to use Bayesian methods to estimate the kinetic parameters for the inactivation kinetics of Cryptosporidium parvum oocysts with chlorine dioxide or ozone which are characterized by the delayed Chick–Watson model, i.e., a lag phase or shoulder followed by pseudo-first-order rate of inactivation. As the length of the lag phase ( CT lag) is not known, Bayesian statistics provides a more accurate approach than traditional statistical methods to fitting the delayed Chick–Watson kinetics. Markov Chain Monte Carlo method is used to estimate CT lag and first-order rate constant values. This method is also used to estimate the minimum CT requirement (with safety factor) for 99% inactivation of C. parvum oocysts.

Effect of turbulent gas–liquid contact in a static mixer on Cryptosporidium parvum oocyst inactivation by ozone

Static mixers may be used to dissolve gaseous ozone in water treatment facilities in order to provide protection against the waterborne parasite Cryptosporidium parvum. The objective of this study was to determine the effect of a brief exposure to turbulent gas–liquid mixing conditions in a static mixer on inactivation of C. parvum oocysts by ozone. Inactivation measured in an ozone contacting apparatus that employed a static mixer for ozone dissolution was compared to predictions based on a previously published kinetic model of C. parvum inactivation by dissolved ozone in gently stirred batch reactors. Although initial contact in the static mixer had no immediate effect on the oocysts, a 20% increase in the rate of inactivation during subsequent contact with dissolved ozone was observed. Increasing the degree of turbulence within the static mixer did not yield additional inactivation. Use of static mixers for dissolution of ozone in drinking water treatment systems may provide limited enhancement of C. parvum inactivation by dissolved ozone.

Assessing UV reactor performance for treatment of finished water

Recently, use of low levels of medium- and low-pressure ultraviolet light for successful inactivation of Cryptosporidium parvum oocysts has generated tremendous excitement in the water industry. Accurate delivery of the target dose, lamp performance, sensor stability and impact of water characteristics are some factors that could impact disinfection efficacy, in turn influencing decisions on application of this technology. To this end, American Water Systems, the largest investor owned water utility in the US, has responded to some of these challenges by ascertaining the long-term feasibility of applying UV for treatment of finished water. A 4 x 1 UV reactor with a 12 inch (0.3 m) diameter was installed after granular activated carbon filtration and was operated with a finished water flow rate of 600 gpm (2,700 L/min). Over a 12-month period, various chemical (THM, HAA, UV254, DOC, TOC, metals, nitrate, nitrites) and physical measurements (lamp voltage, current, sensor measurements) were monitored to define their impact (if any) on the operation of the reactor. MS2 bacteriophage challenge studies were conducted with various lamp configurations and lamp age. These inactivation data demonstrated high levels of correlation with controlled bench scale inactivation data. For C. parvum oocysts, bench scale studies were performed with a modified in vitro infectivity assay using HCT-8 cells, an enhanced infectivity protocol and with either immunofluorescence or quantitative PCR based detection. While both assays indicated increasing infections levels of HCT-8 cells with increasing oocyst inocula, UV treatment of oocysts produced markedly different infectivity responses. Based on the data generated in this study, one in vitro infectivity assay was selected to demonstrate > 3 logs inactivation with low UV doses (5 mJ/cm(20-10 mJ/cm2).

Inactivation of Cryptosporidium parvum oocysts in fresh apple cider by UV irradiation.

This study evaluated the efficacy of UV irradiation on the inactivation of Cryptosporidium parvum oocysts in fresh apple cider. Cider was inoculated with oocysts and exposed to 14.32 mJ of UV irradiation/cm(2). Oocyst viability was assessed with the gamma interferon gene knockout (GKO) mouse and infant BALB/cByJ mouse models. All GKO mice challenged with UV-treated cider demonstrated no morbidity or mortality, and infant BALB/c mice challenged with treated cider were negative for the presence of C. parvum. In contrast, the GKO mice challenged with non-UV-treated inoculated cider died and the parasite was detected in the ileums of all challenged infant mice. This study shows that UV irradiation can be used to inactivate C. parvum in fresh apple cider.

Cryptosporidium parvum oocyst inactivation in three soil types at various temperatures and water potentials

The interaction between soil types, temperature, and soil water potential may have differential effects on the survival of Cryptosporidium parvum oocysts in the terrestrial environment. We examined the effects of three soil types (a silty clay loam, silt loam, and loamy sand), three temperatures (4, 20, and 30 °C), and three soil water potentials (−0.033, −0.5 and −1.5 MPa) on the inactivation kinetics of oocysts. Sentinel chambers were filled with air-dried and sieved soil, brought to the appropriate soil water potential, and inoculated with 2×10 6 freshly purified oocysts. The inoculated chambers were buried in the same bulk soil at the appropriate water potentials and incubated at one of the three temperatures. Triplicate chambers were removed from the bulk soil on days 0, 22, 43, 84 and 156. Sentinel oocysts were extracted, and assayed for potential infectivity by the dye permeability method. Oocysts suspended in sterile distilled water and incubated with the sentinel chambers were used as controls for the effect of temperature. The soil water potentials investigated did not affect oocyst inactivation at any temperature or with any of the three soil types. Rates of oocyst inactivation increased significantly between 4 and 20 °C, but not between 20 and 30 °C with the exception of oocysts incubated in the silty clay loam. Oocyst survival appeared to be significantly greater in the silt loam soil than in the two other soil types when incubated at 20 °C; and at 30 °C oocyst survival was significantly less in the silt clay loam than in the other two soil types. Rates of sentinel oocyst inactivation at all three soil water potentials were significantly lower than the control oocysts in water at the three test temperatures. Thus oocyst survival in soil was not affected by the water potentials between −0.033 and −1.5 MPa; it was affected by soil texture; but temperature appeared to be the factor most affecting oocyst survival. In the critical ambient range of temperature in temperate climates oocysts may survive for months in agricultural soil, and pose a threat to surface waters.

Inactivation of Cryptosporidium parvum oocysts with ozone and free chlorine

The objective of this study is to investigate the synergy involved in the sequential inactivation of C. parvum oocysts with ozone followed by free chlorine at 1–20°C. Primary ozone and free chlorine inactivation curves are characterized by an initial lag-phase, followed by one or two post-lag-phase segments, the first segment at a faster rate than the second, of pseudo-first-order inactivation. The kinetics of primary inactivation with ozone and free chlorine has a relatively strong temperature dependence, and vary both with oocyst lot and oocyst age. Synergy is observed for the sequential inactivation of C. parvum oocysts with ozone/free chlorine. Ozone pre-treatment results in the disappearance of the lag-phase and the occurrence of a secondary free chlorine inactivation curve with generally two pseudo-first-order segments, the first segment at a faster rate than the second. The kinetics of both secondary segments is significantly faster than the post-lag-phase rate of inactivation with free chlorine alone. The temperature dependence for both phases of the secondary free chlorine inactivation kinetics is weaker compared to that for primary inactivation with ozone or free chlorine. As a result, the level of synergy in sequential disinfection with ozone/free chlorine increases with decreasing temperature within the range relevant to drinking water utilities. Good agreement is found between the kinetics determined using the modified in-vitro excystation method of viability assessment and animal infectivity data recently reported in the literature for both primary inactivation with ozone, and sequential disinfection with ozone/free chlorine.

Sequential inactivation of Cryptosporidium parvum oocysts with chlorine dioxide followed by free chlorine or monochloramine

The main objective of this study was to assess the effect of temperature (4–30°C) on the inactivation kinetics of Cryptosporidium parvum oocysts with sequential disinfection schemes involving the use of chlorine dioxide as the primary disinfectant and free or combined chlorine as the secondary disinfectant in synthetic water. The synergy previously reported for sequential inactivation of C. parvum oocysts with ozone/free chlorine or ozone/combined chlorine did not occur when chlorine dioxide was used, instead of ozone, as the primary disinfectant within the temperature range (4–30°C) and the pre-treatment levels investigated. Sequential ozone/chlorine dioxide and chlorine dioxide/ozone experiments revealed that the lower level or absence of synergy for chlorine dioxide/free chlorine and chlorine dioxide/monochloramine was likely the result of chlorine dioxide reacting with oocyst chemical groups that are mostly different from those reacting with ozone, free chlorine, or monochloramine. The CT concept was found to be valid for the primary inactivation kinetics of C. parvum oocysts with chlorine dioxide, thus allowing the use of the simpler CT approach for the development of C. parvum inactivation requirements with chlorine dioxide. General consistency was found between the secondary inactivation kinetics of C. parvum oocysts with free chlorine and monochloramine after chlorine dioxide pretreatment obtained in this study with oocyst viability determined by a modified in vitro excystation method and those reported in the literature for the same sequential disinfection schemes based on an animal infectivity assay.

Chlorine Dioxide Inactivation of C ryptosporidium Parvum in Oxidant Demand-Free Phosphate Buffer

Chlorine dioxide inactivation of Cryptosporidium parvum oocysts was studied at bench-scale in oxidant demand-free 0.05 M phosphate buffer at pH 6, 8, and 11, and temperature from 1°C to 37°C. Animal infectivity using neonatal CD-1 mice was used for evaluation of oocyst infectiousness before and after treatment. Survival curves of the oocysts following treatment declined linearly with the chlorine dioxide Cavgt product. Temperature was critical for C. parvum inactivation, while pH was found not to be a significant factor at pH 6–11. Inactivation kinetics at different temperatures was expressed as a Chick-Watson model with different reaction rate constants adjusted by van't Hoff-Arrhenius relationship. Between 1°C and 37°C, for every 10°C decrease in temperature, the reaction rate constant decreased by a factor of 2.3, corresponding to an activation energy of 54.9 kJ/mol. Design criteria targeting 0.5 to 2.0 log-units of inactivation of C. parvum were developed for different water temperatures and their 90%...

Comparative effectiveness of UV wavelengths for the inactivation of Cryptosporidium parvum oocysts in water

Cryptosporidium parvum oocysts in water were exposed to distinct wavelength bands of collimated beam ultraviolet (UV) radiation across the germicidal UV wavelength range (210-295 nm) that were emitted from a medium pressure (MP) mercury vapour lamp. The dose of UV radiation transmitted though each narrow bandpass filter was measured utilising potassium ferrioxalate actinometry. Oocyst infectivity was determined using a cell culture assay and titre was expressed as an MPN. The log10 inactivation for each band of radiation was determined for a dose of 2 mJ/cm2. Doses from all wavelengths between 250-275 nm resulted in approximately 2 log10 inactivation of Cryptosporidium parvum oocyst infectivity while doses with wavelengths higher and lower than this range were less effective. Because polychromatic radiation from MP UV lamps had about the same germicidal activity between the wavelengths of 250-275 nm for inactivation of oocyst infectivity, there was no unique advantage of MP UV over low pressure (LP) UV except for the simultaneous delivery of a wide range of germicidal wavelengths.

The effect of temperature on the efficacy of ozonation for inactivating Cryptosporidium parvum oocysts

Examination of the effects of water temperature on the inactivation of Cryptosporidium parvum oocysts with ozone, ozonation experiments were conducted in a semi-batch mode with a wide temperature range of 3-30 degrees C. Inactivation was assessed in terms of mice infectivity and in vitro excystation. The temperature dependency of the CT products by a reduction in infectivity of 2 log10 could be described successfully by the Arrhenius equation, 1/CT = 1.086 x 10(18)e-12520/K where CT is the integrated ozone concentration over the contact time (mg/min/L) and K is the Kelvin temperature of water. As for the reduction in viability assessed by the excystation assay, protocol B, the obtained regression equation, 1/CT = 1.802 x 10(18)e-12640/K, was almost identical to that observed for the infectivity. Thus, the CT products required for a 2 log10 reduction in both infectivity and viability increased by an average factor of 4.2 for every 10 degrees C decrease in water temperature. Additionally, our findings suggested that the viability, as determined by protocol B, could substitute for animal infectivity in evaluating the effects of environmental factors on the efficacy of ozonation.

Inactivation of Cryptosporidium parvum oocysts with sequential application of ozone and combined chlorine

Single-step inactivation experiments with ozone and monochloramine revealed the presence of a CT lag followed by pseudo-first order inactivation kinetics. Sequential disinfection experiments with ozone followed by monochloramine revealed that ozone pretreatment resulted in the removal of a more prominent CT lag observed for monochloramine. In addition, the rate of inactivation for ozone-pretreated oocysts was approximately 2.5x greater than that observed for the post-lag phase portion of the monochloramine primary inactivation curve.

Effect of Temperature on Ozone Inactivation of Cryptosporidium parvum in Oxidant Demand-Free Phosphate Buffer

Ozone inactivation of Cryptosporidium parvum oocysts was studied at bench-scale in 0.05 M phosphate buffer at 1 to 37°C, pH 6–8. Animal infectivity using neonatal CD-1 mice was used for evaluation of oocyst infectiousness following treatment. Survival curves of ozone inactivation were characterized by a tail-off effect, with an initial shoulder most evident at low temperature. Temperature was a critical factor for ozone inactivation kinetics with a significant decrease of ozone efficacy at low temperature. Accounting for ozone residual stability at different pH conditions, pH was found to have no significant effect on the activation of C. parvum by ozone. Inactivation kinetics at different temperatures were expressed as an Incomplete gamma Hom model with different reaction rate constants, adjusted for water temperature using the van't Hoff-Arrhenius relationship. Between 1 and 37°C, for every 10°C decrease in the water temperature, the inactivation rate constant decreased by a factor of 2.2, corresponding...

Inactivation of Cryptosporidium parvum Oocysts in Cider by Flash Pasteurization

Cryptosporidium parvum is a well-recognized pathogen of significant medical importance, and cider (apple juice) has been associated with foodborne cryptosporidiosis. This study investigated the effect of flash pasteurization on the viability of contaminant C. parvum oocysts. Cider inoculated with oocysts was heated at 70 or 71.7°C for 5, 10, or 20 s, and oocyst viability was measured by a semiquantitative in vitro infectivity assay. By infecting multiple wells of confluent Madin-Darby bovine kidney cells with serial dilutions of heat-treated oocysts and examining infected cells by indirect fluorescent antibody staining, the most probable number technique was applied to quantify log reduction of oocyst viability. Heating for 10 or 20 s at either temperature caused oocyst killing of at least 4.9 log (or 99.999%), whereas oocyst inactivation after pasteurization for 5 s at 70 and 71.7°C was 3.0 log (99.9%) and 4.8 log (99.998%), respectively. Our results suggested that current practices of flash pasteurization in the juice industry are sufficient in inactivating contaminant oocysts.

Inactivation of cryptosporidium parvum oocysts using medium- and low-pressure ultraviolet radiation

The effect of ultraviolet radiation from low- and medium-pressure mercury arc lamps on Cryptosporidium parvum oocysts was studied using a collimated beam apparatus. Experiments were conducted using parasites suspended in both filtered surface water and phosphate buffered laboratory water. Inactivation of oocysts was measured as reduction in infectivity using a CD-1 neonatal mouse model and was found to be a non-linear function of UV dose over the range of germicidal doses tested (0.8–119 mJ/cm 2). Oocyst inactivation increased rapidly with UV dose at doses less than 25 mJ/cm 2 with two and three log-units inactivation at approximately 10 and 25 mJ/cm 2, respectively. The cause of significant leveling-off and tailing in the UV inactivation curve at higher doses was not determined. Maximum measured oocyst inactivation ranged from 3.4 to greater than 4.9 log-units and was dependent on different batches of parasites. Water type and temperature, the concentration of oocysts in the suspension, and the UV irradiance did not have significant impacts on oocyst inactivation. When compared on the basis of germicidal UV dose, the oocysts were equally sensitive to low- and medium-pressure UV radiation. With respect to Cryptosporidium, both low- and medium-pressure ultraviolet radiation are attractive alternatives to conventional chemical disinfection methods in drinking water treatment.

Sequential Inactivation of CryptosporidiumUsing Ozone Followed by Free Chlorine in Natural Water

Sequential inactivation of Cryptosporidium parvum oocysts in modified natural water using ozone followed by free chlorine was studied. Experiments were conducted at 22°C in modified North Saskatchewan River water at pH 8.5 and pH 6. Animal infectivity using neonatal CD-1 mice was used to measure oocyst viability after treatment. The results showed that the overall inactivation of C parvum using ozone followed by free chlorine in modified NSR water was greater at pH 6 compared; to pH 8.5, but that a synergistic effect was observed at pH 8.5 and not at pH 6. The results also indicated that models developed for ozone inactivation in lab water adequately predicted the inactivation of C. parvum in the modified natural water but models developed for free chlorine in lab water did not give an adequate prediction of the free chlorine inactivation. There were no natural water effects on the inactivation of C. parvum when chemical treatment was not applied.

Inactivation of Cryptosporidium parvum oocysts with ozone and monochloramine at low temperature

The rate of Cryptosporidium parvum inactivation decreased with decreasing temperature (1–20°C) for ozone and for monochloramine applied alone as well as after pre-treatment with ozone. Synergy was observed at all temperatures studied for the ozone/monochloramine sequential disinfection scheme. The synergistic effect was found to increase with decreasing temperature. The inactivation rate with monochloramine after ozone pre-treatment was 5 times faster at 20°C and 22 times faster at 1°C than the corresponding post-lag phase rates of inactivation with monochloramine at these temperatures when no ozone pre-treatment was applied. The CT required for achieving 2-logs of inactivation ranged from 11 400 mg min l −1 at 20°C to 64 600 mg min l −1 at 1°C when monochloramine was applied alone. In contrast, the CT required for an overall sequential inactivation of 2-logs ranged from 721 mg min l −1 at 20°C to 1350 mg min l −1 at 1°C when applying monochloramine after ozone pre-treatment. The presence of excess ammonia in the monochloramine solutions was not responsible for the synergy observed in ozone/monochloramine sequential disinfection.

Sequential inactivation of Cryptosporidium parvum oocysts with ozone and free chlorine

The rate of Cryptosporidium parvum inactivation with free chlorine decreased with increasing pH in the range of 6.0–8.5, consistent with hypochlorous acid being primarily responsible for C. parvum inactivation within this pH range. The rate of sequential inactivation of C. parvum oocysts with free chlorine after pre-ozonation was characterized by an initial rapid decline in viability followed by slower kinetics with respective rates at pH 6.0 of approximately six times and twice that for free chlorine treatment without pre-ozonation under the same conditions. The greatest level of synergy was observed at pH 6; synergy decreased as pH increased until no synergy was observed at pH 8.5. Consistent with hypochlorous acid being the free chlorine species primarily responsible for C. parvum inactivation, within the pH range of 6.0–8.5 the rate of secondary inactivation with free chlorine decreased with increasing pH. Experiments designed to assess the effect of free chlorine concentration on inactivation kinetics provided conclusive evidence for the validity of the CT concept in the case of secondary inactivation of C. parvum oocysts with hypochlorous acid after ozone pre-treatment.

Inactivation of Cryptosporidium parvum oocysts and other microbes in water and wastewater by electrochemically generated mixed oxidants

Alternative disinfectants of water and wastewater are needed because conventional chlorination is ineffective against C. parvum oocysts. Reliable indicators of disinfection efficacy against C. parvum also are needed. Mixedoxidants (MO) electrochemically generated from brine were evaluated in batch disinfection experiments for inactivation of C. parvum oocysts and Cl. perfringensspores in both oxidant demand-free (ODF) water and treated wastewater. Coliphage MS2 and Escherichia coli B were also tested under some conditions. C. parvum oocyst infectivity was quantified by cell culture assay, and the dyes DAPI (4′,6-diamidino-2-phenylindole) and propidium iodide (PI) were used to assess oocyst viability in wastewater experiments. In treated wastewater dosed with 10–13 mg/L MO, inactivation after 90 minutes was about 3 log10 for C. parvum and about 2.5 log10 for Cl. perfringens spores; MS2 and E. coli were rapidly inactivated by > 5 log10. In ODF water, a 4 mg/L dose of MO inactivated ∼3 log10 of C. parvum oocysts and ∼1.5 log10 of Cl. perfringens spores. Inactivation of C. parvum oocysts and Cl. perfringensspores was less extensive at a lower MO dose of 2 mg/L. The use of DAPI and PI to determine viability of oocysts treated with MO did not correlate with, and greatly overestimated, cell culture infectivity. At practical doses and contact times, MO disinfection of water and wastewater achieves appreciable inactivation of both C. parvum oocysts and Cl. perfringens spores. Cl. perfringens spores reliably indicated oocyst inactivation by MO, but E. coli and coliphage MS2 were inactivated much too rapidly to indicate C. parvum inactivation.

Inactivation of Cryptosporidium parvum oocysts with chlorine dioxide

The effect of solution pH and temperature on the rate of inactivation of Iowa strain Cryptosporidium parvum ( C. parvum) oocysts with chlorine dioxide was determined. Oocyst viability was assessed with a modified in vitro excystation method consistent with animal infectivity data. Inactivation curves were characterized by a lag phase followed by Chick–Watson first order kinetics. The rate of inactivation was essentially the same at pH 6 and 8. CT requirements at pH 10 were approximately 20–30% lower than those at pH 6–8. The magnitude of the lag phase and the inactivation rate constant were both found to obey Arrhenius law for the experimental temperature range of 4–30°C. CT requirements for C. parvum oocyst inactivation were found to increase by an average factor of approximately 3.4 for every 10°C decrease in temperature.