Porphyrins are photosensitizers and may be applicable in situations where viral inactivation is required, as for in vitro inactivation of nonenveloped viruses in blood components or in other aqueous media. No study has examined the efficacy of porphyrin inactivation on human pathogens such as hepatitis A virus (HAV) in plasma or other liquids. Experiments were conducted to evaluate the effect of synthetic porphyrins on HAV in porphyrin-containing human plasma and phosphate-buffered saline exposed to long-wavelength (365 nm) UV light. Inactivation of bacteriophage MS2 (MS2) also was determined in some trials. Solutions containing cationic, anionic or amphiphilic porphyrins irradiated with an average light dose of 4.3 J/cm(2) for 90 min resulted in >3 log(10) (>99.9%) to >4 log(10) (>99.99%) inactivation of both HAV and MS2. Viral inactivation may have been greater than observed because the limits of detection of the assay had been reached. Under ambient lighting conditions, none of the porphyrins was mutagenic in the Ames assay and only the congener with the longest chain-length, tetrakis (N-[n-hexadecyl]-4-pyridiniumyl) porphyrin, was appreciably toxic to mammalian cells. Disinfection by photoactivated synthetic porphyrins therefore can offer an effective and relatively safe approach to removal of nonenveloped viruses from aqueous media.
Hurricane Floyd and other storms in 1999 caused widespread and extensive flooding of eastern North Carolina and environmental contamination with fecal wastes from municipal wastewater and livestock operations. Because wastewater contains high levels of pathogenic micro-organisms, principal health risks to humans from flooding are consumption of crops grown in fecally contaminated soil and ingestion of contaminated water. Flood waters polluted with microbial and other contaminants also may be detrimental to the health of livestock and plant crops. In the present study, agricultural soils impacted by flood waters were analyzed for bacterial and viral indicators of fecal contamination. Total coliforms, fecal coliforms, Escherichia coli, spores of Clostridium perfringens, and both male specific (F+) and somatic coliphages were recovered from soil and assayed in liquid culture media. A number of samples were positive for the presence of fecal coliforms, E. coli, and coliphages, indicating the presence of human or animal feces. Most samples were positive for total coliforms, and almost all samples contained high levels of Cl. perfringens spores. The levels of Cl. perfringens spores were significantly (P < 0.001) higher in flooded soil (post-Hurricane Floyd) compared to pre-flood soil. Persistent fecal contamination of soil, as demonstrated by the high levels of Cl. perfringens spores, suggests the need for additional or alternative measures to protect crop-growing areas, including prospective microbiological monitoring and improved protection of watersheds from incidents capable of releasing fecal material.
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.
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