Efficacy of Peracetic Acid and Sodium Hypochlorite against SARS-CoV-2 on Contaminated Surfaces.

SARS-CoV-2 is primarily a respiratory virus that can potentially be transmitted through fomites. Sodium hypochlorite (NaOCl) and peracetic acid (PAA) are widely used disinfectants on surfaces in diverse settings such as hospitals and food production facilities. The objectives of this study were to investigate the virucidal efficacy of NaOCl and PAA against SARS-CoV-2 using the ASTM standard methods. In the suspension assay, NaOCl and PAA (5, 50, and 200 ppm) were tested against SARS-CoV-2 in the presence/absence of soil load after 1 min of contact time. In the carrier assay, NaOCl and PAA were tested at 200, 400, 600, and 1,000 ppm for 1 min and 200 and 1,000 ppm for 5 and 10 min. Stainless steel (SS) and high-density polyethylene (HDPE) disks were used as carriers. The virus was suspended in soil load and the disinfectants were prepared in 300 ppm of hard water. Virus quantification was done by TCID50 assay using Vero-E6 cell line. NaOCl and PAA were effective (> 3 log reduction in infectious virus) at 50 ppm in the absence of soil load. However, in the presence of soil load, 200 ppm was required for > 3 log reduction in virus infectivity. In contrast, NaOCl and PAA at 200 ppm and with a 1-min contact time were not effective against SARS-CoV-2 on either SS or HDPE surfaces. PAA at 200 ppm for 10 min was effective against SARS-CoV-2 on SS and HDPE surfaces, whereas NaOCl required 1,000 ppm for 10 min to be effective against SARS-CoV-2 on both surfaces. IMPORTANCE In the context of the COVID-19 pandemic, the World Health Organization (WHO) recommended the use of chlorine-based products such as sodium hypochlorite (NaOCl) at 1,000 ppm for a minimum of 1 min to disinfect environmental surfaces. However, this recommendation was not based on validated studies on the actual SARS-CoV-2 itself. In fact, over half of the chemical disinfectants, including many peracetic acid products, listed in EPA List N were approved based on "kills a harder-to-kill pathogen" without further validation on SARS-CoV-2. Research on SARS-CoV-2 is restricted to BSL3 laboratories and the urgency of tackling the pandemic might explain the lack of studies on the actual virus. Our results show that the WHO recommendation of 1 min contact time with 1,000 ppm NaOCl is not effective against SARS-CoV-2 on surfaces. Also, our results indicate that PAA is effective against SARS-CoV-2 on surfaces and can be used as safer and more environmentally friendly alternative to NaOCl at a lower concentration.

Application of denaturing gradient gel electrophoresis (DGGE) for assessing fecal pollution sources at a recreational beach.

We investigated the effectiveness of Escherichia coli community fingerprinting for identifying fecal pollution sources impacting a recreational beach. E. coli in water collected from the beach, nearby creek and storm sewer outfall were enumerated using membrane filtration, while E. coli communities were characterized following polymerase chain reaction analysis and denaturing gradient gel electrophoresis (DGGE) fingerprinting. Analysis of E. coli densities to determine the contributions of the creek and storm sewer during dry weather was inconclusive. However, DGGE fingerprinting indicated that the creek E. coli communities had a greater impact on the beach community composition (80-95% similarity), than on storm sewer communities (41-64%). Following rainfall events, E. coli communities in the creek were at least 93% similar to those at the beach, while the similarity of the outfall and beach communities varied from 72 to 96%. Furthermore, E. coli communities at the beach were more similar to creek communities than to storm sewer communities after the first 2 h and 48 h following the onset of rainfall, and of comparable similarity following 24 h of rainfall, suggesting transient contributions from the storm sewer. DGGE analysis of E. coli communities provided evidence that the creek was a consistent source of E. coli to the beach, while the storm sewer was a transient source.

Detection and differentiation of staphylococcal contamination of clinical surfaces using denaturing gradient gel electrophoresis.

The detection and identification of staphylococci from the environment of at-risk patients can be an important step in determining the role of the environment in hospital-acquired infections. Current methods that are used to identify these pathogens are either limited in their capabilities, expensive and/or labour intensive. We developed a denaturing gradient gel electrophoresis (DGGE) analysis protocol for the detection and identification of staphylococci that takes advantage of species-specific polymorphisms in the gene that encodes elongation factor Tu (tuf). The protocol was optimised by performing polymerase chain reaction (PCR)-DGGE analysis on DNA isolated from pure cultures of 27 different staphylococcal species. This resulted in the separation of the PCR products into 19 different band positions, including unique positions for important species such as Staphylococcus aureus, S. hominis, S. lugdunensis, S. warneri, S. capitis, S. caprae and S. saprophyticus. Application of the method was demonstrated by swabbing 15 clinical surfaces in an isolation room occupied by a patient before and after routine cleaning. PCR-DGGE analysis of tuf showed that despite cleaning efforts, the surfaces remained contaminated with several species of staphylococci, including S. aureus, S. epidermidis, S. lugdunensis, S. hominis, S. haemolyticus and S. simulans. We conclude that DGGE of tuf represents a promising technique for the detection, characterisation and monitoring of mixed assemblages of staphylococci in the healthcare environment.