Mic H. Stewart

Polymorphisms in the β-tubulin gene of Cryptosporidium parvum differentiate between isolates based on animal host but not geographic origin

Polymerase chain reaction primers were designed to target a region of the Cryptosporidium parvum beta-tubulin gene spanning an intron. Amplification products contained 11 polymorphic positions, representing a sequence divergence of 1.8%, which discriminated between isolates of C. parvum found solely in humans (genotype 1) and those found in humans and animals (genotype 2). Seven of the polymorphic sites were located outside of the intron and the polymorphism between isolates was readily demonstrated by HaeIII restriction digestion. However, all of the sequences from genotype 1 human-derived oocysts isolated in the United States and Australia were conserved. Also, there were no sequence differences between bovine isolates obtained from both continents. Therefore, isolates could not be differentiated based on geographic source of origin.

Development of a Rapid Detection Procedure for Ctyptosporidium, Using In Vitro Cell Culture Combined with PCR

A detection, viability, and infectivity assay was developed for Cryptosporidiurn parvum. Oocysts or excysted sporozoites were inoculated onto monolayers of CaCo‐2 cells grown on chamber slides. C. parvum infection was monitored by three methods: a) application of a fluorescein‐labeled anti‐sporozoite antibody; b) PCR of a heat‐shock protein gene fragment; and c) detection of mRNA from the heat‐shock protein gene by RT‐PCR.

Modeling the impact of body-contact recreation on pathogen concentrations in a source drinking water reservoir

Abstract A modeling study was conducted to evaluate the impact of body-contact recreation (e.g., water skiing, jet skiing, swimming) on pathogen concentrations in a source drinking water reservoir under construction in eastern Riverside County in Southern California. A hybridized Monte Carlo-finite segment model was used to predict pathogen concentrations in the reservoir resulting from pathogen inputs associated with shed fecal material and accidental fecal releases (AFRs). Monte Carlo techniques were incorporated into the finite segment model to define characteristics about individual recreators which affect pathogen loading to the reservoir (e.g., infection, pathogen shedding rate, location). Results of simulations are provided in the form of cumulative distribution and probability density functions derived from uncertainty analyses. The model predicted considerable spatial and temporal variability in pathogen concentrations within the reservoir, with elevated levels of Cryptosporidium , rotavirus, and poliovirus in the epilimnion during periods of high recreational use. Predicted Giardia concentrations were lower than the other pathogens. Hypolimnetic concentrations of all pathogens were generally 1–3 orders of magnitude lower than the overlying epilimnetic concentrations. Model results also suggest that field sampling will underestimate the mean, range and variance of pathogen concentrations in the reservoir. The model was further modified to include a particle tracking scheme to allow for transport of aggregated fecal material. Results from simulations using this approach demonstrate a potential for high pathogen loads due to body-contact recreation periodically reaching treatment plants.

Production and Removal of Assimilable Organic Carbon Under Pilot-Plant Conditions Through the Use of Ozone and PEROXONE

Pilot-plant studies were conducted in two source waters to determine the effects of predisinfection with ozone alone and with a combination of hydrogen peroxide and ozone (PEROXONE) on the production of assimilable organic carbon (AOC) compounds. Colorado River water (CRW) and State project water (SPW) from Northern California were treated with ozone alone at applied dosages ranging from 1 to 4 mg/L and with PEROXONE at hydrogen peroxide/ozone (H2O2/O3) ratios of 0.05, 0.10, 0.20, and 0.30. Ozonation of CRW with applied dosages of 1.0,2.0, and 4.0 mg/L increased AOC concentrations from 70μg C/L to 275, 350, and 224 μg C/L, respectively. Ozonation of SPW with an applied dosage of 4.0 mg/L elevated AOC concentrations from 70 to 522μg C/L.

Pilot-Plant-Scale Ozone and PEROXONE Disinfection of Giardia muris Seeded into Surface Water Supplies

Abstract PEROXONE is an advanced oxidation process for water treatment which is generated by combining ozone and hydrogen peroxide. In this study, surface water supplies were seeded with viable Giardia muris cysts and disinfected by ozone and PEROXONE in the pretreatment columns of a 22.7 L/min pilot plant. Inactivation was examined in two source waters under conditions of varying applied ozone doses (0.5–4.0 mg/L), with and without the addition of hydrogen peroxide; at increasing contact times (3, 6, 9, and 12 min); and with and without added high turbidity (10 and 50 NTU). Approximately 108 viable G. muris cysts were injected into the ozone contactor-column influent. The cysts were collected at the ozone contactor-column effluent and concentrated, and viability was then determined using in vitro excystation.