Log10 Plaque-forming Units Research Articles

Evaluation of an automated far ultraviolet-C light technology for decontamination of surfaces and aerosolized viruses in bathrooms

BackgroundAerosols generated during toilet flushing are a potential source for transmission of viral and bacterial pathogens in bathrooms. However, manual decontamination of bathrooms after each use is not feasible.MethodsWe tested the efficacy of a wall-mounted far ultraviolet-C (UV-C) light technology that only delivers far UV-C when people are not present for decontamination of surfaces and aerosolized viral particles in an unoccupied hospital bathroom. A quantitative disk carrier test method was used to test efficacy against organisms on steel disk carriers placed in 9 sites in the bathroom with an exposure time of 45 min and 2 h; Clostridioides difficile spores were also exposed for 24 h. Efficacy against aerosolized bacteriophage MS2 was tested with a 45-minute exposure.ResultsThe far UV-C technology reduced methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), Candida auris, and bacteriophage MS2 on steel disk carriers by ≥ 1.2 log10 (range, 1.2 to 4.2 log10) at all test sites after 2 h of exposure. The technology reduced C. difficile spores by < 1 log10 after 2 h exposure, but 4 of 9 test locations had ≥ 2 log10 reductions after 24 h exposure. Aerosolized bacteriophage MS2 was reduced by 4 log10 plaque-forming units in 45 min.ConclusionsThe far UV-C light technology could potentially be useful for automated decontamination of air and surfaces in bathrooms in healthcare and community settings.

Household dishwashing detergents efficiently inactivate HSV-1, but not the non-enveloped viruses HAV and MNV, while all viruses were removed from glass by manual scrubbing

This study investigated the stability of herpes simplex virus 1 (HSV-1), hepatitis A virus (HAV) and murine norovirus (MNV) on glass, their inactivation by three household dishwashing detergents and during a glass rinsing process. After drying on glass and storage in darkness, virus titers decreased by 4.7, 5.5 and 4.5 log10 plaque forming units (PFU) in 21, 28 and 14 days for HSV-1, HAV and MNV, respectively. Daylight storage resulted in shorter survival times. While HAV and MNV titers were not significantly affected by 60 s incubation in dishwashing detergents at up to 43 °C, HSV-1 titer decreased significantly by > 4 log10 PFU/mL at the same conditions with each detergent used. After cleaning artificially contaminated drinking glasses with a manual glass scrubbing device, HSV-1, HAV and MNV titers decreased significantly by 4.4 ± 0.1 to 5.3 ± 0.0 log10 PFU. Our study shows, that all tested viruses can remain infectious after drying on glass up to several weeks. Household dishwashing detergents efficiently inactivate the enveloped virus HSV-1, but not the non-enveloped viruses HAV and MNV. The applied manual glass rinsing procedure efficiently removed all three viruses from drinking glasses, probably due to the combination of chemical and mechanical treatments.

Refined semi-lethal aerosol H5N1 influenza model in cynomolgus macaques for evaluation of medical countermeasures

Highly pathogenic avian influenza A H5N1 viruses cause high mortality in humans and have pandemic potential. Effective vaccines and treatments against this threat are urgently needed. Here, we have refined our previously established model of lethal H5N1 infection in cynomolgus macaques. An inhaled aerosol virus dose of 5.1 log10 plaque-forming unit (pfu) induced a strong febrile response and acute respiratory disease, with four out of six macaques succumbing after challenge. Vaccination with three doses of adjuvanted seasonal quadrivalent influenza vaccine elicited low but detectable neutralizing antibody to H5N1. All six vaccinated macaques survived four times the 50% lethal dose of aerosolized H5N1, while four of six unvaccinated controls succumbed to disease. Although vaccination did not protect against severe influenza, vaccinees had reduced respiratory dysfunction and lower viral load in airways compared to controls. We anticipate that our macaque model will play a vital role in evaluating vaccines and antivirals against influenza pandemics.

Efficacy of a far-ultraviolet-C light technology for continuous decontamination of air and surfaces.

A wall-mounted, far-ultraviolet-C light technology reduced aerosolized bacteriophage MS2 by >3 log10 plaque-forming units within 30 minutes. Vegetative bacterial pathogens on steel disk carriers in the center of the room were reduced by >3 log10 after 45 minutes of exposure, but Candida auris and Clostridioides difficile spores were not.

Persistence and transfer of Tulane virus in a microgreen cultivation system

Microgreens are niche salad greens which have increased in popularity among consumers in recent years. Due to similarities with sprouts and leafy greens—both attributed to numerous foodborne disease outbreaks—characterization of the food safety risks associated with microgreen production is warranted. The present study aimed to determine the fate and persistence of a human norovirus (HuNoV) surrogate, Tulane virus (TV), within a microgreen production system. Initially, the persistence of TV in two types of microgreen soil-free cultivation matrix (SFCM)—BioStrate® (biostrate) and peat—was determined. On day 0, water containing 7.6 log PFU of TV was applied to SFCM in growing trays, and the trays were maintained under microgreen growth conditions. TV persisted throughout the 10-day observation in biostrate and peat with overall reductions of 3.04 and 1.76 log plaque forming units (PFU) per tray, respectively. Subsequently, the transfer of TV to microgreen edible tissue was determined when planted on contaminated SFCM. Trays containing each type of SFCM were pre-inoculated with 7.6 log PFU of TV and equally divided into two areas. On day 0, sunflower (SF) or pea shoot (PS) seeds were planted on one-half of each tray, while the other half was left unplanted to serve as a control. The microgreens were harvested on day 10, and SFCM samples were collected from planted and unplanted areas of each tray. No TV were detected from the edible portion of either type of microgreen, yet TV were still present in the SFCM. TV concentrations were significantly lower in the root-containing planted area compared with the unplanted area for both biostrate (P = 0.0282) and peat (P = 0.0054). The mean differences of TV concentrations between unplanted and planted areas were 1.22 and 0.51 log PFU/g for biostrate and peat, respectively. In a subsequent investigation, TV transfer from day 7 inoculated SFCM to microgreens edible portion was not detected either. Overall, this study characterized the viral risk in a microgreen production system, which will help to understand the potential food safety risk related to HuNoV and to develop preventive measures.

Sensitivity of RNA viral nucleic acid-based detection of avian influenza virus, Newcastle disease virus, and African horse sickness virus on flinders technology associates card using conventional reverse-transcription polymerase chain reaction.

The flinders technology associates (FTA) card is a cotton-based cellulose membrane impregnated with a chaotropic agent that inactivates infectious microorganisms, lyses cellular material, and fixes DNA and/or RNA within the fiber matrix. However, little is known about the effectiveness of these cards for detecting RNA viruses in animals. This study aimed to evaluate the sensitivity of RNA virus detection using conventional reverse-transcription polymerase chain reaction (RT-PCR) on FTA cards. A highly virulent Newcastle disease virus (NDV) and an avian influenza virus (AIV) with low pathogenicity were propagated using chicken embryonic eggs. Three days after inoculation, the allantoic fluid was harvested, stored at -80°C, and the stock virus was tested for virus titration. African horse sickness virus (AHSV) was obtained from a live attenuated vaccine that was dissolved and stored at -80°C. For sample preparation, each stock virus was 10-fold serially diluted and each dilution was inoculated onto an FTA card, followed by drying in a Class II safety cabinet. Both the stock virus and infected FTA card were genomically isolated using an extraction kit, FTA purification kit, and extraction kit with Tris-EDTA (TE) buffer. The target genome was then detected by one-step RT-PCR for NDV and AIV, and two-step RT-PCR for African horse sickness, including gel electrophoresis for the detection of specific nucleic acids. The detection limit of stock AIV was compared on FTA cards, using the FTA purification kit, and with TE buffer with an extraction kit. The corresponding results were 1.47, 1.17, and 2.18 log10 EID50, respectively, while for NDV the results were 4.13, 4.83, and 4.84 log10 ELD50. Finally, detection limit of stock AHSV and AHSV on the FTA card extracted using TE buffer with an extraction kit were 4.30 and 4.01 log10 plaque-forming units, respectively. This study demonstrated that the detection limit or sensitivity of all tested RNA viruses on FTA cards did not differ when compared with those of the stock virus and in both methods for RNA isolation on FTA cards. These cards are suitable for collecting and transporting samples infected with RNA viruses, particularly AIV, NDV, and AHSV. Flinders technology associates cards also provide hazard-free samples, a reliable source of RNA for molecular characterization, and sufficient quantity for diagnostic applications based on nucleic acid-based detection.

Ultraviolet dosage and decontamination efficacy were widely variable across 14 UV devices after testing a dried enveloped ribonucleic acid virus surrogate for SARS-CoV-2.

Aims: The dosages and efficacy of 14 ultraviolet (UV) decontamination technologies were measured against a SARS-CoV-2 surrogate virus that was dried onto different materials for laboratory and field testing. Methods and results: A live enveloped, ribonucleic acid (RNA) virus surrogate for SARS-CoV-2 was dried on stainless steel 304 (SS304), Navy Top Coat-painted SS304 (NTC), cardboard, polyurethane, polymethyl methacrylate (PMMA), and acrylonitrile butadiene styrene (ABS) materials at > 8.0 log10 plaque-forming units (PFU) per test coupon. The coupons were then exposed to UV radiation during both laboratory and field testing. Commercial and prototype UV-emitting devices were measured for efficacy: four handheld devices, three room/surface-disinfecting machines, five air disinfection devices, and two larger custom-made machines. UV device dosages ranged from 0.01 to 729 mJ cm−2. The antiviral efficacy among the different UV devices ranged from no decontamination up to nearly achieving sterilization. Importantly, cardboard required far greater dosage than SS304. Conclusion: Enormous variability in dosage and efficacy was measured among the different UV devices. Porous materials limit the utility of UV decontamination. Significance and impact of the study: UV devices have wide variability in dosages, efficacy, hazards, and UV output over time, indicating that each UV device needs independent technical measurement and assessment for product development prior to and during use.

Optimized Protocols for the Propagation and Quantification of Infectious Murine Hepatitis Virus (MHV-A59) Using NCTC Clone 1469 and 929 Cells.

Murine hepatitis virus (MHV) is a non-human pathogen betacoronavirus that is evolutionarily and structurally related to the human pathogenic viruses SARS-CoV, MERS-CoV, and SARS-CoV-2. However, unlike the human SARS and MERS viruses, MHV requires a biosafety level 2 laboratory for propagating and safe handling, making it a potentially suitable surrogate virus. Despite this utility, few papers discussed the propagation and quantification of MHV using cell lines readily available in biorepositories making their implementations not easily reproducible. This article provides protocols for propagating and quantifying MHV-A59 using the recommended NCTC clone 1469 and clone 929 cell lines from American Type Culture Collection (ATCC). More specifically, the methods detail reviving cells, routine cell passaging, preparing freeze stocks, infection of NCTC clone 1469 with MHV and subsequent harvesting, and plaque assay quantification of MHV using NCTC clone 929 cells. Using these protocols, a BSL-2 laboratory equipped for cell culture work would generate at least 6.0 log plaque-forming units (PFU) per mL of MHV lysate and provide an optimized overlay assay using either methylcellulose or agarose as overlays for the titration of infectious virus particles. The protocols described here are intended to be utilized for persistence and inactivation studies of coronaviruses.

Laboratory and Field Evaluations of a Commercially Available Real-Time Loop-Mediated Isothermal Amplification Assay for the Detection of West Nile Virus in Mosquito Pools.

Although the specific cDNA amplification mechanisms of reverse-transcriptase polymerase chain reaction (RT-PCR) and RT loop-mediated isothermal amplification (RT-LAMP) are very different, both molecular assays serve as options to detect arboviral RNA in mosquito pools. Like RT-PCR, RT-LAMP uses a reverse transcription step to synthesize complementary DNA (cDNA) from an RNA template and then uses target-specific primers to amplify cDNA to detectable levels in a single-tube reaction. Using laboratory-generated West Nile virus (WNV) samples and field-collected mosquito pools, we evaluated the sensitivity and specificity of a commercially available WNV real-time RT-LAMP assay (Pro-AmpRT™ WNV; Pro-Lab Diagnostics, Inc., Round Rock, Texas) and compared the results to a validated real-time RT-PCR assay. Laboratory generated virus stock samples containing ≥ 2.3 log10 plaque-forming units (PFU)/ml and intrathoracically inoculated mosquitoes containing ≥ 2.4 log10 PFU/ml produced positive results in the Pro-AmpRT WNV assay. Of field-collected pools that were WNV positive by real-time RT-PCR, 74.5% (70 of 94) were also positive by the Pro-AmpRT WNV assay, resulting in an overall Cohen's kappa agreement of 79.4% between the 2 tests. The Pro-AmpRT WNV assay shows promise as a suitable virus screening tool for vector surveillance programs provided agencies are aware of its characteristics and limitations.

No Evidence of rVSV-Ebola Virus Vaccine Replication or Dissemination in the Sand Fly Phlebotomus papatasi.

Following vaccination with the live attenuated, recombinant vesicular stomatitis virus Indiana serotype Ebola virus (rVSV-EBOV) vaccine, persons may exhibit a transient vaccine-associated viremia. To investigate the potential for Old World sand flies to transmit this vaccine following feeding on a viremic person, we fed laboratory-reared Phlebotomus papatasi an artificial blood meal containing 7.2 log10 plaque-forming units of rVSV-EBOV. Replication or dissemination was not detected in the body or legs of any P. papatasi collected at seven (n = 75) or 15 (n = 75) days post-feed. These results indicate a low potential for rVSV-EBOV to replicate and disseminate in P. papatasi, a species whose geographic distribution ranges from Morocco to southwest Asia and as far north as southern Europe.

Modeling the Stability of SARS-CoV-2 on Personal Protective Equipment (PPE)

.We modeled the stability of SARS-CoV-2 on personal protective equipment (PPE) commonly worn in hospitals when carrying out high-risk airway procedures. Evaluated PPE included the visors and hoods of two brands of commercially available powered air purifying respirators, a disposable face shield, and Tyvek coveralls. Following an exposure to 4.3 log10 plaque-forming units (PFUs) of SARS-CoV-2, all materials displayed a reduction in titer of > 4.2 log10 by 72 hours postexposure, with detectable titers at 72 hours varying by material (1.1–2.3 log10 PFU/mL). Our results highlight the need for proper doffing and disinfection of PPE, or disposal, to reduce the risk of SARS-CoV-2 contact or fomite transmission.

Safety and Antiviral Effects of Nebulized PC786 in a Respiratory Syncytial Virus Challenge Study.

BackgroundPC786 is a nebulized nonnucleoside respiratory syncytial virus (RSV) polymerase inhibitor designed to treat RSV, which replicates in the superficial layer of epithelial cells lining the airways.MethodsFifty-six healthy volunteers inoculated with RSV-A (Memphis 37b) were randomly dosed with either nebulized PC786 (5 mg) or placebo, twice daily for 5 days, from either 12 hours after confirmation of RSV infection or 6 days after virus inoculation. Viral load (VL), disease severity, pharmacokinetics, and safety were assessed until discharge. RSV infection was confirmed by reverse-transcription quantitative polymerase chain reaction with any positive value (intention-to-treat infected [ITT-I] population) or RSV RNA ≥1 log10 plaque-forming unit equivalents (PFUe)/mL (specific intention-to-treat infection [ITT-IS] population) in nasal wash samples.ResultsIn the ITT-I population, the mean VL area under the curve (AUC) was lower in the PC786 group than the placebo group (274.1 vs 406.6 log10 PFUe/mL × hour; P = .0359). PC786 showed a trend toward reduction of symptom score and mucous weight. In ITT-IS (post hoc analysis), the latter was statistically significant as well as VL AUC (P = .0126). PC786 showed an early time to maximum plasma concentration, limited systemic exposure, and long half-life and consequently a 2-fold accumulation over the 5-day dosing period. PC786 was well tolerated.ConclusionsNebulized PC786 demonstrated a significant antiviral effect against RSV, warranting further clinical study.Clinical Trials RegistrationClinicalTrials.gov: NCT03382431; EudraCT: 2017-002563-18.

Hot, Humid Air Decontamination of Aircraft Confirmed That High Temperature and High Humidity Are Critical for Inactivation of Infectious, Enveloped Ribonucleic Acid (RNA) Virus.

Aims: To develop infectious (live/dead) enveloped virus test indicators and response surface methodology (RSM) models that evaluate survival of an enveloped ribonucleic acid (RNA) virus on contaminated aircraft materials after exposure to hot, humid air (HHA).Methods and Results: Enveloped RNA bacteriophage Phi6 (Φ6) was dried on wiring insulation, aircraft performance coating (APC), polypropylene, and nylon at ≥ 8 log10 plaque-forming units (PFU) test coupon−1. Only 2.4 log10 inactivation was measured on APC at 70°Celsius (°C), 5% relative humidity (RH) after 24 h. In contrast, HHA RSM models showed a 90% probability of a 7 log10 inactivation at ≥63°C, 90% RH after 1 h, and decontamination kinetics were similar across different materials. HHA decontamination of C-130 and C-17 aircraft showed >7 log10 and ≥5.9 log10 inactivation of enveloped virus on 100 and 110 test indicators, respectively, with a 1-h treatment, excluding ramp-up and ramp-down times.Conclusions: Enveloped RNA virus test indicators were successfully developed, lab tested for HHA decontamination, analyzed for RSM, and field-tested in aircraft demonstrations.Significance and Impact of the Study: The utility of HHA decontamination was demonstrated after inactivating enveloped RNA virus on aircraft with a 1-h HHA treatment within aircraft temperature and RH limits.

What’s in a Handshake? Exploring the Best Form of Greeting to Prevent Hand to Hand Spread of Viruses

Background: Respiratory and enteric viruses are highly contagious pathogens that can be spread by contaminated hands and surfaces. We hypothesized that alternatives to handshake greetings that reduce the time and surface area of hand contact would be associated with decreased transfer of viral particles. Methods: In a simulation of hand-contact greetings, volunteers (N = 22) used a keyboard contaminated with the benign bacteriophage MS2 and then performed a handshake and fist bump with additional volunteers. To assess viral transfer, hands were cultured for MS2, and plaque-forming units (PFU) were compared for the different types of hand contact. Additional simulations (N = 10) were conducted to compare viral transfer with the fist bump versus a cruise tap greeting (ie, a modified fist bump involving single knuckle contact). Results: The handshake greeting resulted in significantly greater transfer of MS2 than the fist bump (1.31 vs 0.54 log10 PFUs, P &lt; .001) (Fig. 1A), but the frequency of transfer of virus was high for both greetings (91% transfer by handshake vs 59% by fist bump). The cruise-tap greeting did not result in reduced transfer of viral particles in comparison to the fist bump (Fig. 1B), and the frequency of transfer remained high (70%). Conclusions: The fist-bump and cruise-tap greetings could potentially reduce transmission of viruses in comparison to the handshake, but transfer occurred frequently, even with these greeting methods. To eliminate hand-to-hand transmission of respiratory and enteric viruses, alternative greeting methods that do not involve physical contact are needed.Funding: NoneDisclosures: None

Embryotoxic impact of Zika virus in a rhesus macaque in vitro implantation model†.

Zika virus (ZIKV) infection is associated with adverse pregnancy outcomes in humans, and infection in the first trimester can lead to miscarriage and stillbirth. Vertical and sexual transmissions of ZIKV have been demonstrated, yet the impact of infection during the initial stages of pregnancy remains unexplored. Here we defined the impact of ZIKV on early embryonic and placental development with a rhesus macaque model. During in vitro fertilization (IVF), macaque gametes were inoculated with a physiologically relevant dose of 5.48log10 plaque-forming units (PFU) of Zika virus/H.sapiens-tc/PUR/2015/PRVABC59_v3c2. Exposure at fertilization did not alter blastocyst formation rates compared to controls. To determine the impact of ZIKV exposure at implantation, hatched blastocysts were incubated with 3.26log10, 4.26log10, or 5.26log10 PFU, or not exposed to ZIKV, followed by extended embryo culture for 10days. ZIKV exposure negatively impacted attachment, growth, and survival in comparison to controls, with exposure to 5.26log10 PFU ZIKV resulting in embryonic degeneration by day 2. Embryonic secretion of pregnancy hormones was lower in ZIKV-exposed embryos. Increasing levels of infectious virus were detected in the culture media post-exposure, suggesting that the trophectoderm is susceptible to productive ZIKV infection. These results demonstrate that ZIKV exposure severely impacts the zona-free blastocyst, whereas exposure at the time of fertilization does not hinder blastocyst formation. Overall, early stages of pregnancy may be profoundly sensitive to infection and pregnancy loss, and the negative impact of ZIKV infection on pregnancy outcomes may be underestimated.

Wheat Strip Effects on Microbial Transport Following Variable Applications of Beef Cattle Manure

Abstract. Vegetative filter strips (VFS) consisting of perennial vegetation have been successfully used to reduce the transport of contaminants in runoff from land application areas. The effectiveness of a winter wheat strip, which may be more acceptable to producers, in reducing microbial transport was examined in this study. A 1.4 m wheat strip was used to allow direct comparison with experimental results obtained in previous studies. Beef cattle manure was applied to 0.75 m wide by 4.0 m long plots established on an Aksarben silty clay loam located in southeast Nebraska. Manure was added at rates required to meet the 0- 1-, 2-, or 4-year phosphorus requirement for corn. The transport of selected microbes was measured for three 30 min simulated rainfall events separated by 24 h intervals. The narrow wheat strip did not significantly reduce counts of any of the measured microbes. The application of manure to meet the 4-year P requirement resulted in and enterococci loads that were significantly greater than the 1-year P requirement. Rainfall simulation run significantly affected measurements of phages, total coliforms, , and enterococci, with measurements during the three runs varying from 9.35 to 10.9 log plaque-forming units (PFU) ha-1, from 11.5 to 12.1 log colony-forming units (CFU) ha-1, from 12.1 to 12.5 log CFU ha-1, and from 11.1 to 11.4 log CFU ha-1, respectively. The transport of was found to be significantly correlated to selected nutrient loads and electrical conductivity of runoff. The presence of narrow wheat strips did not reduce microbial loads in runoff. Keywords: Bacteria, Cattle manure, E. coli, Filter strips, Land application, Manure management, Manure runoff, Microbial, Microorganisms, Runoff.

Optimizing Propagation of Staphylococcus aureus Infecting Bacteriophage vB_SauM-phiIPLA-RODI on Staphylococcus xylosus Using Response Surface Methodology.

The use of bacteriophages for killing pathogenic bacteria is a feasible alternative to antibiotics and disinfectants. To obtain the large quantities of phages required for this application, large-scale production of bacteriophages must be optimized. This study aims to define conditions that maximize the phage yield of the virulent and polyvalent staphylococcal bacteriophage vB_SauM-phiIPLA-RODI in broth culture, using the food-grade species Staphylococcus xylosus as the host strain to reduce the risk of growing massive quantities of pathogenic bacteria and therefore, to ensure the safety of the final phage stock. The effect of four variables, namely initial bacterial concentration (5.66–8.40 log10 colony-forming unit (CFU)/mL), initial phage concentration (5–8 log10 plaque-forming unit (PFU)/mL), temperature (21–40 °C) and agitation (20–250 rpm), on phage yield (response) was studied by using response surface methodology (RSM). Successive experimental designs showed that agitation did not significantly impact phage yield, while temperature did have a significant effect, with 38 °C being the optimum for phage propagation. The results allowed the design of a model to describe phage yield as a function of the initial bacterial and phage concentrations at fixed agitation (135 rpm), and optimum temperature (38 °C). The maximum experimental phage yield obtained was 9.3 log10 PFU/mL, while that predicted by the model under the optimized conditions (7.07 log10 CFU/mL initial bacterial population and 6.00 log10 PFU/mL initial phage titer) was 9.25 ± 0.30 log10 PFU/mL, with the desirability of 0.96. This yield is comparable to that obtained when the phage was propagated on the original host, Staphylococcus aureus. Bacteriophage phiIPLA-RODI showed the same host range and very similar biofilm removal ability regardless of the staphylococcal species used for its propagation. The results presented in this study show the suitability of using a food-grade strain of S. xylosus for the propagation of S. aureus infecting phages and the application of RSM to define the optimal propagation conditions.

Reptiles and Amphibians as Potential Reservoir Hosts of Chikungunya Virus.

Chikungunya virus is an emerging arbovirus of significant human-health concern. Little is known about its sylvatic cycle, including whether ectothermic vertebrates are permissive to infection. In this study, individuals from ten species of reptiles and amphibians were inoculated with chikungunya virus and samples of blood were tested to characterize viremia and seroconversion. Viremia was not detected in cane toads, house geckos, or American alligators, but most of the green iguanas, red-eared sliders, ball and Burmese pythons, leopard frogs, Texas toads, and garter snakes developed viremia. Peak virus titers in serum of up to 4.5, 4.7, and 5.1 log10 plaque-forming units per milliliter were observed for garter snakes, ball pythons, and Texas toads, respectively. These results add to those of other studies that have suggested a possible role for ectothermic vertebrates in the ecology of arbovirus maintenance and transmission in nature.

Reducing Dissemination of Viruses from Computer Touchscreens through Patient Hand Hygiene and an Automated Ultraviolet-C Touchscreen Disinfection Device

BackgroundComputer touchscreens in patient waiting areas are a potential source for spread of viral and bacterial pathogens in healthcare facilities. Patient hand hygiene is recommended to prevent acquisition of pathogens on hands.MethodsWe tested the efficacy of patient hand hygiene alone or in combination with a novel automated ultraviolet-C (UV-C) touchscreen disinfection device for reduction in dissemination of the nonpathogenic non-enveloped virus bacteriophage MS2 from contaminated touchscreens. Subjects randomized to control, alcohol hand sanitizer (10 or 30 second application), UV-C (30-second cycle), or alcohol hand sanitizer plus UV-C contacted 4 sites routinely used during operation of the touchscreen and then fingerpads were cultured. The log plaque-forming units (PFU) recovered were compared for each group. Patients were observed to determine the frequency of hand hygiene after use of a touchscreen in a waiting area.ResultsAs shown in the figure, the mean log10PFU of bacteriophage MS2 recovered from fingerpads was significantly reduced by each of the interventions (P < 0.001). However, only the combination of hand hygiene and UV-C was effective in completely preventing virus transfer. Of 20 patients observed in a waiting area, 0 (0%) used hand sanitizer that was available adjacent to the touchscreen.ConclusionOur results suggest that use of alcohol hand sanitizer alone or in combination with an automated UV-C touchscreen disinfection device could reduce transmission of viruses from contaminated touchscreens. There is a need for education of patients on the importance of hand hygiene after contact with touchscreens.Figure.Efficacy of hand hygiene (HH) and/or UV-C in reducing recovery of bacteriophage MS2 from fingers after contact with contaminated touchscreensDisclosures All authors: No reported disclosures.

Minimum infectious dose for chikungunya virus in Aedes aegypti and Ae. albopictus mosquitoes

ABSTRACTUnderstanding the ability of the chikungunya virus (CHIKV) to be transmitted by Aedes vectors in the Americas is critical for assessing epidemiological risk. One element that must be considered is the minimum infectious dose of virus that can lead to transmission following the extrinsic incubation period. This study aimed to determine the minimum infection rate for the two Aedes species studied. The results revealed that doses as low as 3.9 log10 plaque-forming units per mL (pfu/mL) of an Asian genotype CHIKV strain can lead to transmission by Ae. albopictus, and doses of at least 5.3 log10 pfu/mL from the same strain are needed for transmission from Ae. aegypti. These low infecting doses suggest that infected individuals may be infectious for almost the entire period of their viremia, and therefore, to prevent further cases, measures should be taken to prevent them from getting bitten by mosquitoes during this period.