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A Surface Coating that Rapidly Inactivates SARS-CoV-2

  • Saeed Behzadinasab
    Saeed Behzadinasab
    Department of Chemical Engineering and Center for Soft Matter and Biological Physics, Virginia Tech, Blacksburg, Virgina 24061, United States
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    • , 
  • Alex Chin
    Alex Chin
    School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China
    More by Alex Chin
    Orcidhttp://orcid.org/0000-0002-6556-9092
    • , 
  • Mohsen Hosseini
    Mohsen Hosseini
    Department of Chemical Engineering and Center for Soft Matter and Biological Physics, Virginia Tech, Blacksburg, Virgina 24061, United States
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    • , 
  • Leo Poon*
    Leo Poon
    School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China
    HKU-Pasteur Research Pole, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, China
    *Email: [email protected]
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    • , and 
  • William A. Ducker*
    William A. Ducker
    Department of Chemical Engineering and Center for Soft Matter and Biological Physics, Virginia Tech, Blacksburg, Virgina 24061, United States
    *Email: [email protected]
    More by William A. Ducker
    Orcidhttp://orcid.org/0000-0002-8207-768X
Cite this: ACS Appl. Mater. Interfaces 2020, 12, 31, 34723–34727
Publication Date (Web):July 13, 2020
https://doi.org/10.1021/acsami.0c11425
Copyright © 2020 American Chemical Society
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Abstract

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SARS-CoV-2, the virus that causes the disease COVID-19, remains viable on solids for periods of up to 1 week, so one potential route for human infection is via exposure to an infectious dose from a solid. We have fabricated and tested a coating that is designed to reduce the longevity of SARS-CoV-2 on solids. The coating consists of cuprous oxide (Cu2O) particles bound with polyurethane. After 1 h on coated glass or stainless steel, the viral titer was reduced by about 99.9% on average compared to the uncoated sample. An advantage of a polyurethane-based coating is that polyurethane is already used to coat a large number of everyday objects. Our coating adheres well to glass and stainless steel as well as everyday items that people may fear to touch during a pandemic, such as a doorknob, a pen, and a credit card keypad button. The coating performs well in the cross-hatch durability test and remains intact and active after 13 days of being immersed in water or after exposure to multiple cycles of exposure to the virus and disinfection.

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  Note

This article is made available via the ACS COVID-19 subset for unrestricted RESEARCH re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

1. Introduction

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COVID-19 has caused widespread human morbidity and mortality as well as disruption to our economy and our way of life. The disease is spread through the virus, SARS-CoV-2, which is known to remain viable on some solids for periods of up to 1 week.(1,2) One potential route of human infection occurs when a person comes into contact with a solid that is contaminated with the virus (a fomite), so one method of reducing transmission would be to reduce the period of human vulnerability to infection by reducing the period of viability of SARS-CoV-2 on solids. In this report, we describe the fabrication and testing of an anti-SARS-CoV-2 coating. Our ultimate goal is to produce a coating that (i) inactivates the virus quickly, (ii) can be coated on many solids, and (iii) is sufficiently robust that it retains virucidal potency during consumer use. Such a film could be used on many household, commercial, medical, and manufacturing surfaces, such as doorknobs, credit card buttons, and cell phone covers. These are objects that a person might touch only minutes after deposition of respiratory droplets. Our hope is that widespread use of such a coating could reduce transmission of SARS-CoV-2 and also reduce fear of touching objects.
At our starting point, it was unknown which chemical groups inactivated SARS-CoV-2. We tested three films. Two of the films were monolayers of cationic polymer: cationic groups are known to inactivate other viruses(3,4) (and bacteria(3−5)), and the mechanism is believed to be that the high density of charge disrupts the self-assembly of the pathogen. These films did not have a significant effect. We chose cuprous oxide (Cu2O) as the active ingredient of the third film. Prior work demonstrated that SARS-CoV-2 had a short period of viability on Cu metal,(1) and the surface layer of Cu metal readily oxidizes to Cu2O.(6) Cu2O has shown activity against two other viruses: a bacteriophage by exposure to unbound particles(7) and hepatitis C by exposure of the virus to suspended nanoparticles.(8)
Cuprous oxide is used as a pesticide and is rated as toxicity category III by the Environmental Protection Agency (EPA), which means that it is slightly toxic and slightly irritating.(9) It is also used as a marine antifoulant, and it is known to have an adverse effect on marine organisms.(10,11) However, a review of clinical and animal studies of textiles containing copper oxide by Borkow reports no effects on human skin.(12) For copper pesticide products, more generally, according to an 2009 EPA report,(9) “Current available literature and studies do not indicate any systemic toxicity associated with copper exposure.... There are no residential or occupational risks of concern resulting from exposure to copper products”. Likewise, an immune response to copper is rare, whereas contact with copper in coins, intrauterine devices, and other applications is common.(12)
In this work, we describe the time course of inactivation of SARS-CoV-2 on Cu2O particles bound into a thin (10–16 μm) film with a commercial polyurethane (PU) (Figure 1). The commercial polyurethane is already formulated to be hard-wearing and easy to apply to many materials. Our results show that the Cu2O/Pu coating inactivates the virus very quickly: viable viral counts drop by an average of about 99.9% in 1 h. The film is robust and retains its potency to SARS-CoV-2 after multiple cycles of exposure to the virus followed by disinfection.

Figure 1

Figure 1. Cross-section view of the Cu2O/polyurethane film.

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2. Materials and Methods

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2.1. Materials

Cu2O particles (Chem Copp HP III Type UltraFine-5) were purchased from American Chemet Corporation. The chemical composition and particle size distribution information are described in Table S1 and Figure S1. Polyurethane (Miniwax, fast drying polyurethane, clear satin) was purchased from Lowe’s Home Improvement store. The following chemicals were purchased from Sigma Aldrich: (3-glycidyloxypropyl)trimethoxysilane (GOPTS) (>98%) and poly(diallyldimethylammonium chloride) (PDADMAC) (average Mw = 400,000–500,000, 20 wt% in water). Polyallylamine (PA) in freebase form (Mw = 15,000, 15% in water) was purchased from Polysciences. The following items were obtained from VWR: glass slides (25 × 75 × 1 mm), toluene (ACS grade), ethanol 200 PROOF, nitric acid (70%, ACS grade), and sodium chloride (ACS grade). Stainless steel 301 shim (thickness, 0.25 mm; McMaster Carr) was cut into 13 × 13 mm pieces, washed with soap and water, and rinsed with water. Water was purified by a Barnstead EASY pure II unit.

2.2. Fabrication of the Films

The Cu2O/PU coating was fabricated as follows. A very thin layer of PU was applied to a glass slide using a sponge and then left to dry for approximately 8 min to allow partial curing of the polymer. At this time, the film showed only slight or zero marking when touched with a gloved hand. Cu2O (10%) in ethanol suspension was sonicated for 3 min, and then, 1 mL was applied to the PU film and left to partially dry for about 5 min at room temperature. The film was then heated in an oven at 120 °C for 2 h to finish the cure, forcefully blown with compressed nitrogen gas, washed thoroughly with DI water, and dried with a stream of nitrogen gas. At this point, the film had a high advancing contact angle for water, approximately 130°. The glass film was broken into ≈12 × 12 mm pieces. Each piece of glass or stainless steel was then cleaned with argon plasma to remove excess polyurethane (see Figure S2). For the plasma cleaning, the sample was placed in vacuum, twice-purged in argon gas for 10 min, and then exposed to argon plasma for 3 min at 100 W and <200 mTorr. After plasma cleaning, the film was wetted by water, but the advancing water contact angle was recovered after 1 day. This is the state in which SARS-CoV-2 inactivation was tested, and all other tests were performed unless otherwise specified. The films showed no visible change after immersion in water for 7 weeks or ethanol for 1 month. When we replaced the oven treatment with 24 h of cure, the outer layer of the film was less stable but a robust layer remained on the solid (see the Supporting Information).
The PA film was prepared, as described previously.(5) Briefly, the glass was rinsed three times with DI water, soaked in 70% ethanol for 15 min, and rinsed three times with DI water. Subsequently, the glass was immersed in 6 M nitric acid for 20 min and then thoroughly rinsed in DI water. The glass was then exposed to oxygen plasma, treated with pure GOPTS at 37 °C for 60 min, and then with 15% PA in water (pH = 11) and left to react at 75 °C for 36 h in a closed container. Subsequently, the films were rinsed with DI water (10 times) to remove unattached polymer chains and dried with nitrogen gas.(5) For the PDADMAC surface, the cleaned glass was immersed in 1 mM PDADMAC/10 mM NaCl for 3 h and then gently rinsed in DI water and dried with nitrogen gas.(13)

2.3. Characterization of the Cu2O Films

An SEM image of the film in cross section is shown in Figure 1, and a plan view is shown in Figure S2 along with images of a film before plasma treatment. As a result of fabrication from a polydisperse particle distribution, the final film has a distribution of particle sizes and is rough; the thickness ranged from 10 to 16 μm. Contact angles were measured using a First Ten Angstroms FTA125. Water contact angles of the film 1 week after fabrication were advancing (120°), receding (<10°), and sessile in the range of 80–120°. The chemical composition of the film was investigated by X-ray photoelectron spectroscopy (XPS; PHI VersaProbe III with a monochromatic Al Kα source of 1486.6 eV) as well as electron-dispersive X-ray spectroscopy (EDX; Bruker Quantax) and scanning electron microscopy (SEM; FEI Quanta 600 FE-ESEM). Results confirmed the chemical identity of the Cu2O particles in the film and are contained in the Supporting Information, Figure S3 and Table S2. The robustness of the film was measured by the cross-hatch test ASTM D3359 Method B and peel test ASTM D1876.

2.4. Characterization of the Cationic Polymer Films

The PA and PDADMAC film water contact angles were < 10°, and the chemical compositions and XPS (Figures S4 and S5) were consistent with the chemical composition.

2.5. Inactivation of SARS-CoV-2

Biological testing methods are described in Chin et al.’s prior work.(2) Briefly, before testing with SARS-CoV-2, the film was disinfected with 70% ethanol in water and air-dried at 37 °C overnight, unless otherwise stated. SARS-CoV-2 was isolated from the index case of Hong Kong. The stock virus was prepared by Vero E6 cells cultured in Dulbecco’s modified Eagle medium supplemented with 2% fetal bovine serum and 1% v/v penicillin–streptomycin at 37 °C with 5% CO2. A 5 μL test droplet containing 6.2 × 107 (7.8 log unit) TCID50/mL SARS-CoV-2 was placed on the test solid at 60–70% humidity and 22–23 °C and was observed to dry in about 15 min for more hydrophilic samples and about 30 min for more hydrophobic samples. After a prescribed time, the solid was soaked in 300 μL of viral transport medium (Earle’s balanced salt solution supplemented with 0.5% (w/v) bovine serum albumin and 0.1% (w/v) glucose, pH 7.4) at room temperature (≈22 °C) to elute the virus. The eluted virus was titrated by a 50% tissue culture infective dose (TCID50) assay in Vero E6 cells.(14,15) In brief, confluent Vero E6 cells on 96-well plates were infected with serially diluted virus in quadruplicates. The infected cells were incubated at 37 °C with 5% CO2. On day 5 post-infection, the cells were examined for a cytopathic effect. The TCID50/mL is the dilution that caused a cytopathic effect in 50% of treated Vero E6 cell cultures (N = 4 per each dilution; Reed–Muench method(16)). Three independent tests were done at each condition, except for the data in Figure S8 where there were six independent measurements. Error residuals were approximately normally distributed after a log transformation (Figure S6), so all averages were calculated from the log of the titers. Because reductions in titer were very large, we make use of the “log reduction” terminology of biology:(1)(2)For example, a 99.9% reduction is the same as a 3-log reduction.

3. Results and Discussion

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3.1. The Cu2O/PU Coating Rapidly Inactivates SARS-CoV-2

Inactivation of the virus by the Cu2O/PU coating is swift and dramatic, typically a reduction of 99.9% of the viral titer in 1 h (Figure 2). The virus is slowly inactivated even on glass or stainless steel, so in eq 1, we always use as our basis of comparison the titer on the uncoated sample at the same time point. Clearly, the numerical reductions would be even greater if we were to compare to the viral titer at time zero.

Figure 2

Figure 2. Time course of viable titer of SARS-CoV-2 on solids with and without a coating of cuprous oxide microparticles bound with polyurethane (Cu2O/PU). Note that the vertical axis is on a log10 scale. Data is shown for coated glass and coated stainless steel. Individual circular data points represent each independent measurement, and the × symbol represents the mean of the log of independent measurements. The detection limit was 90 TCID50/mL (shown with a dotted line). Experimental results where the virus was not detected are plotted at 90 TCID50/mL and are included in the average as 90 TCID50/mL. SARS-CoV-2 is inactivated much more rapidly on the coated surface than on the bare surface.

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On glass, the SARS-CoV-2 titer was below the detection limit after 1 h for all three samples and at all further times, so the reduction is at least 99.98% (Table 1). The 95% confidence interval is the range greater than 99.5%. On stainless steel, the reduction was 99.90% after 1 h. The reductions in titer on glass and stainless steel are very similar because the virus interacts with the coating, not the underlying material. Films were fabricated by hand, so some variations may be due to differences in sample preparation as well as in testing. Numerical values of data points are in Tables S3–S10. The effects of the polyurethane-only coating were not resolved (p > 0.05 at 1 h), demonstrating that Cu2O is a necessary ingredient in the film. Figure S7 shows the time course of the viral titer on polyurethane.
Table 1. Average Reduction of the SARS-CoV-2 Titer on a Cu2O/PU-Coated Solid Compared to the Titer on an Uncoated Solid. Comparison is at 1 h
conditioncomparison% reductionlog reduction95% CIap-valuecfigure no.
Cu2O/PU coating on glassglass>99.98>3.6499.955 × 10–42
Cu2O/PU coating on stainless steelstainless steel99.902.9798.518 × 10–32
PU coating on glassglass100.04–1640.22S7
Cu2O/PU on glass, stored 13 days under waterglassb99.963.3999.568 × 10–4S10
Cu2O/PU glass, high contact angleglass99.892.9799.222 × 10–6S8
Cu2O/PU glass, 5× disinfectionglass99.892.9599.794 × 10–83
a

95% confidence limit lower bound. Upper bound set at 100%. Calculated for one-tail, assuming heteroscedastic.

b

Comparison sample not stored under water.

c

p-values for Student’s t test calculated for one-tail, assuming heteroscedastic.

Several mechanisms for the biocidal activity of cuprous oxide compounds have been proposed, and the mechanism may vary according to the microbe. Fujimori et al. reported that the mechanism was dissolution of Cu+ followed by production of reactive oxygen species (ROS), e.g., OH·. These ROS would then go on to degrade viral proteins.(17,18) In contrast, Sunada et al. reported that a direct contact with Cu2O with a virus was required for virucidal activity,(7) and Hang et al. reported that Cu2O can inhibit infection by preventing virus entry.(8) The inhibitory mechanism of Cu2O on SARS-CoV-2 is beyond the scope of this study, but further investigation on this topic is warranted.
It is interesting to note that the time course of inactivation appeared to be affected by the wettability of the Cu2O/PU film. The first batch of coating that we made produced high sessile contact angles of viral culture medium, >90° (possibly due to some polyurethane on the surface), whereas later batches had lower angles, ≈45° (precise measurement not available in the BSL-3 lab.). XPS and SEM characterizations are from the later batches. When the angle was high, we were able to detect a small amount of virus at 1 h on some replicates (see Figure S8.) We rationalize this effect as follows. A lower contact angle leads to a higher contact area between the viral suspension and the coating and/or faster transport within the liquid that was able to partially penetrate the film.
The coating is easy to apply to everyday objects because the first stage of the coating procedure is to paint with a commercial polyurethane, which has been formulated to adhere to a variety of surfaces. The Supporting Information (Figure S9) shows images of various items—a doorknob, a credit card reader “Enter” button, a pen, and a supermarket cart handle—coated in the Cu2O/PU film.
We also note that there was no cytopathic effect to the Vero E6 cells after exposure to the viral transport medium that had been in contact with the Cu2O/PU film, consistent with low toxicity of the film.

3.2. The Cu2O/PU Film Remains Potent after Multiple Exposures to Virus or Storage under Water

To determine the potency of the film for inactivating virus after multiple exposure to virus, we applied a 5 μL droplet containing SARS-CoV-2 to a Cu2O/PU-coated surface and allowed the droplet and then its dry residue to stand for 24 h to ensure inactivation of the virus. The coated solid was then cleaned by soaking in 70% ethanol. After air drying, another 5 μL droplet containing virus was applied on the same spot of the surface. These procedures were repeated five times, and then, we tested the ability of the coating to inactivate SARS-CoV-2, as described in the Materials and Methods section. The film remained potent for inactivating SARS-CoV-2: after five exposure/disinfection cycles, the film still reduced the viral titer by about 99.89% in 1 h (see Figure 3), which is similar to the potency of the same films prior the disinfection cycles (99.88%) (see Figure S8).

Figure 3

Figure 3. Time course of viable titer of SARS-CoV-2 on glass coated in Cu2O/PU that was subjected to five cycles of exposure to SARS-CoV-2 plus soaking in 70% ethanol. The uncoated glass was also subjected to the disinfection cycles.

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Samples were also stored under water for 13 days and then tested. The coating retained its virucidal ability for SARS-CoV-2 (see Figure S10).

3.3. The Cu2O/PU Film is Robust

The Cu2O/PU film is not noticeably scratched when handled with tweezers, although it can be removed by heavy, deliberate scratching. Five samples of the Cu2O/PU film on stainless steel were scratched through to the steel with a razor blade in an 11 × 11, 1 mm square grid, and then, an attempt was made to remove each of the 100 small pieces of the coating with tape (ASTM cross-hatch test ASTM D3359). For three independent samples, on average, only 2.4 of the 100 squares were affected with a standard deviation of 0.8, corresponding to a 4B rating in the ASTM standard (see Figure S11). When the tape was used to pull off the top layer of particles from an unscratched sample (a variant of the ASTM D1876 peel test(19)), there was a 0.25% loss of coverage (p = 0.007, six samples); visual inspection of the images showed that only a few particles were removed (see Figure S12). These results indicate that the film has durability.

3.4. A Cationic Polymer Coating did not Speed Inactivation of SARS-CoV-2

Two cationic polymer films were examined: a film of PA (mainly primary amines) and a film of PDADMAC (quaternary ammonium). Both films were designed such that the polymer retained flexibility. The PA was tethered by covalent linkages, and prior force measurements showed that polymer chains could extend up 200 nm from the solid in solution.(5) PDADMAC chains could extend up to 10 nm from the solid.(13) This chain flexibility was designed to assist contact with a larger area of the curved virus. The cationic coatings did not, however, lead to greater inactivation of the virus for the first 24 h for PDADMAC or for the first 48 h for PA (see Figures S13 and S14).

4. Conclusions

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A porous coating of Cu2O/PU causes a swift and dramatic reduction of the infective titer of SARS-CoV-2. After 1 h, a typical reduction is 99.9% compared to the uncoated sample. The coating retains its potency after multiple exposures to virus followed by washing with 70% ethanol in water or after immersion in water for 13 days. The coating retains its mechanical integrity after being cut with a razor blade. Amine and ammonium-based polymers did not inactivate the virus under the conditions tested.

Supporting Information

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The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsami.0c11425.

  • Chemical composition of Cu2O particles (Table S1), elemental composition of the Cu2O/PU film measured by EDS (Table S2), virus titration data (Tables S3–S10), alternative methods for preparing the Cu2O/ PU film, particle size distribution of Cu2O particles (Figure S1), SEM Images of the Cu2O/PU coating before and after plasma cleaning (Figure S2), XPS results for Cu2O/PU (Figure S3), XPS results for the PA polymer films (Figure S4), XPS results for the PDADMAC polymer films (Figure S5), histogram of residuals for the virus titer (Figure S6), viral titer for the polyurethane film (Figure S7), viral titer for the high-contact angle sample (Figure S8), images of various items coated in Cu2O/PU (Figure S9), viral titer for samples stored under water for 13 days (Figure S10), cross-hatch test results for Cu2O/PU on stainless steel (Figure S11), peel test results (Figure S12), viral titer for PA (polyallylamine) (Figure S13), and viral titer for PDADMAC (Figure S14) (PDF)

Terms & Conditions

Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

Author Information

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  • Corresponding Authors
    • Leo Poon - School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China;  HKU-Pasteur Research Pole, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, China;  Email: [email protected]
    • William A. Ducker - Department of Chemical Engineering and Center for Soft Matter and Biological Physics, Virginia Tech, Blacksburg, Virgina 24061, United States;  Orcidhttp://orcid.org/0000-0002-8207-768X;  Email: [email protected]
  • Authors
    • Saeed Behzadinasab - Department of Chemical Engineering and Center for Soft Matter and Biological Physics, Virginia Tech, Blacksburg, Virgina 24061, United States
    • Alex Chin - School of Public Health, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China;  Orcidhttp://orcid.org/0000-0002-6556-9092
    • Mohsen Hosseini - Department of Chemical Engineering and Center for Soft Matter and Biological Physics, Virginia Tech, Blacksburg, Virgina 24061, United States
  • Author Contributions

    S.B. and A.C. contributed equally.

  • Notes

    The authors declare no competing financial interest.

Acknowledgments

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The authors thank Dr. Stephen McCartney for capturing the SEM images and acknowledging the use of Electron Microscopy facilities within the Nanoscale Characterization and Fabrication Laboratory at Virginia Polytechnic Institute. We also thank Dr. Xu Feng for capturing the XPS spectra and acknowledge the use of the Surface Analysis Laboratory in the Department of Chemistry at Virginia Tech, which is supported by the National Science Foundation under grant no. CHE-1531834. We also thank Matthew Ducker for help with statistical analysis, Dr. Dave Dillard for advice on the cross-hatch test, and Dr. Ayman Karim for the loan of an argon cylinder. Cu2O chemical composition and size distribution were provided by American Chemet Corporation. This work was supported by the National Science Foundation under grant no. CBET 1902364, the Health and Medical Research Fund (COVID190116), and the National Institute of Allergy and Infectious Diseases (contract HHSN272201400006C).

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    Painting a glass slide with branched or linear N,N-dodecyl methylpolyethylenimines (PEIs) and certain other hydrophobic PEI derivs. enables it to kill influenza virus with essentially a 100% efficiency (at least a 4-log redn. in the viral titer) within minutes, as well as the airborne human pathogenic bacteria Escherichia coli and Staphylococcus aureus. For most of the coating polyions, this virucidal action is shown to be on contact, i.e., solely by the polymeric chains anchored to the slide surface; for others, a contribution of the polyion leaching from the painted surface cannot be ruled out. A relationship between the structure of the derivatized PEI and the resultant virucidal activity of the painted surface has been elucidated.
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  5. 5
    Iarikov, D. D.; Kargar, M.; Sahari, A.; Russel, L.; Gause, K. T.; Behkam, B.; Ducker, W. A. Antimicrobial Surfaces Using Covalently Bound Polyallylamine. Biomacromolecules 2014, 15, 169– 176,  DOI: 10.1021/bm401440h
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    Antimicrobial Surfaces Using Covalently Bound Polyallylamine
    Iarikov, Dmitri D.; Kargar, Mehdi; Sahari, Ali; Russel, Lauren; Gause, Katelyn T.; Behkam, Bahareh; Ducker, William A.
    Biomacromolecules (2014), 15 (1), 169-176CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)
    We investigated the antimicrobial properties of the cationic polymer polyallylamine (PA) when covalently bonded to glass. The objective was to obtain a robust attachment, yet still allow extension of the polymer chain into soln. to enable interaction with the bacteria. The PA film displayed strong antimicrobial activity against Staphylococcus epidermidis, Staphylococcus aureus, and Pseudomonas aeruginosa, which includes both Gram-pos. and Gram-neg. bacteria. Glass surfaces were prepd. by a straightforward two-step procedure of first functionalizing with epoxide groups using 3-glycidoxypropyltrimethoxy silane (GOPTS) and then exposing to PA so that the PA could bind via reaction of a fraction of its amine groups. The surfaces were characterized using XPS and Fourier transform IR spectroscopy to verify the presence of the polymer on the surface, zeta potential measurements to est. the surface charge of the films, and at. force microscopy to det. the extension of the polymer chains into soln. Antimicrobial properties of these coatings were evaluated by spraying aq. suspensions of bacteria on the functionalized glass slides, incubating them under agar, and counting the no. of surviving cell colonies.
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    Sunada, K.; Minoshima, M.; Hashimoto, K. Highly Efficient Antiviral and Antibacterial Activities of Solid-State Cuprous Compounds. J. Hazard. Mater. 2012, 235-236, 265– 270,  DOI: 10.1016/j.jhazmat.2012.07.052
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    Highly efficient antiviral and antibacterial activities of solid-state cuprous compounds
    Sunada, Kayano; Minoshima, Masafumi; Hashimoto, Kazuhito
    Journal of Hazardous Materials (2012), 235-236 (), 265-270CODEN: JHMAD9; ISSN:0304-3894. (Elsevier B.V.)
    Several solid-state cuprous compds., including cuprous oxide (Cu2O), sulfide (Cu2S), iodide (CuI), and chloride (CuCl), have highly efficient antiviral activities, whereas those of solid-state silver and cupric compds. are markedly lower. On a Cu2O-loaded glass substrate, for example, the infectious activity of bacteriophages was reduced by 5-orders of magnitude within 30 min and by 3-orders of magnitude within 1 h for bacteria. In contrast, the infectious activities of both phages and bacteria were not markedly reduced on CuO-loaded substrates within a similar time frame. To det. the origin of this inhibitory activity, this study investigated the effects of reactive oxygen species (ROS), leached copper ions, and the solid-state compd. itself against bacteriophages, and concluded that infectious activity is lost following direct contact with the solid-state surface of cuprous compds., but not ROS or copper ions. The Cu2O adsorbed and denatured more proteins than CuO, which suggests the difference of the inhibitory activity between Cu2O and CuO.
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    Hang, X.; Peng, H.; Song, H.; Qi, Z.; Miao, X.; Xu, W. Antiviral Activity of Cuprous Oxide Nanoparticles against Hepatitis C Virus in Vitro. J. Virol. Methods 2015, 222, 150– 157,  DOI: 10.1016/j.jviromet.2015.06.010
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    Antiviral activity of cuprous oxide nanoparticles against Hepatitis C Virus in vitro
    Hang, Xiaofeng; Peng, Haoran; Song, Hongyuan; Qi, Zhongtian; Miao, Xiaohui; Xu, Wensheng
    Journal of Virological Methods (2015), 222 (), 150-157CODEN: JVMEDH; ISSN:0166-0934. (Elsevier B.V.)
    Small mol. inhibitors in combination with or without interferon have improved sustained antiviral responses against Hepatitis C Virus (HCV) infection. Nonetheless, resistance to these inhibitors is expected to emerge rapidly due to the high mutation rate of the virus. Thus, new antiviral drugs, in combination with currently available therapies, are urgently needed to treat HCV infection. In the present study, we evaluated the antiviral efficacy of cuprous oxide nanoparticles (CO-NPs) against HCV in the HCVcc/Huh7.5.1 cell culture system. CO-NPs were able to significantly inhibit the infectivity of HCVcc at a non-cytotoxic concn. In addn., CO-NPs inhibited the entry of HCV pseudoparticle (HCVpp), including genotypes 1a, 1b, and 2a, while no effect on HCV replication was obsd. Further time-of-addn. expt. indicated that CO-NPs blocked HCV infection both at the attachment and entry stages. In conclusion, we report that CO-NPs can act as an anti-HCV agent by targeting the binding of infectious HCV particles to hepatic cells and the virus entry into the cells. These findings suggest that CO-NPs may have novel roles in the treatment of patients with chronic hepatitis C.
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    EPA Reregistration Eligibility Decision (RED) for Coppers; https://www3.epa.gov/pesticides/chem_search/reg_actions/reregistration/red_G-26_26-May-09.pdf 2009.
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    Chen, D.; Zhang, D.; Yu, J. C.; Chan, K. M. Effects of Cu2O Nanoparticle and CuCl2 on Zebrafish Larvae and a Liver Cell-line. Aquat. Toxicol. 2011, 105, 344– 354,  DOI: 10.1016/j.aquatox.2011.07.005
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    Effects of Cu2O nanoparticle and CuCl2 on zebrafish larvae and a liver cell-line
    Chen, Dongshi; Zhang, Dieqing; Yu, Jimmy C.; Chan, King Ming
    Aquatic Toxicology (2011), 105 (3-4), 344-354CODEN: AQTODG; ISSN:0166-445X. (Elsevier B.V.)
    The extensive uses of nanomaterials have caused many concerns of their potential hazards to the aquatic environments. As partial dissoln. of metal nanoparticles may occur, it is important to study the toxic effects of nanoparticles and det. the no observable effect levels (NOELs) and lowest observable effect levels (LOELs) of these materials in water by using biomarker genes' expression in zebrafish (Danio rerio). In this study, the toxic effects of Cu2O nanoparticle (NP) on zebrafish larvae and zebrafish liver cell-line (ZFL) were evaluated by detg. their 96 h LC50 values (zebrafish larvae: 242.4 ppb; ZFL: 110 ppm), which was less toxic than CuCl2 (zebrafish larvae: 85.73 ppb; ZFL: 23.04 ppm). However, zebrafish larvae are sensitive to both Cu2O NP and CuCl2. We also examd. the effects of elevated Cu2O NP and CuCl2 on the expression of several Cu related genes in zebrafish larvae by real-time quant. PCR . It was found that Cu2O NP and CuCl2 induced the mRNA levels of metallothionein (MT), Cu/Zn superoxide dismutase (Cu/Zn SOD), metal regulatory transcription factor 1 (MTF1) and copper transporters, ATP7A and 7B, but down-regulated the mRNA levels of glutathione sulfur transferase (GST). Interestingly, the inductions of MT, ATP7A and ATP7B in the Cu2O NP exposure groups were much higher than that of the CuCl2 exposure groups, and resulted in higher Cu accumulation in the Cu2O NP exposure groups. Furthermore, as detd. by MT, ATP7A and ATP7B gene expression, the NOELs of CuCl2 and Cu2O NP were 11 ppb and 30 ppb whereas the LOELs of CuCl2 and Cu2O NP were 43 ppb and 121 ppb, resp.
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    Amara, I.; Miled, W.; Slama, R. B.; Ladhari, N. Antifouling Processes and Toxicity Effects of Antifouling Paints on Marine Environment A Review. Environ. Toxicol. Pharmacol. 2018, 57, 115– 130,  DOI: 10.1016/j.etap.2017.12.001
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    Antifouling processes and toxicity effects of antifouling paints on marine environment. A review
    Amara, Intissar; Miled, Wafa; Ben Slama, Rihab; Ladhari, Neji
    Environmental Toxicology and Pharmacology (2018), 57 (), 115-130CODEN: ETOPFR; ISSN:1382-6689. (Elsevier B.V.)
    A review. The prodn. infrastructure in aquaculture invariably is a complex assortment of submerged components with cages, nets, floats and ropes. Cages are generally made from polyamide or high d. polyethylene (PEHD). All of these structures serve as surfaces for biofouling. However, cage nets and supporting infrastructure offer fouling organisms thousands of square meters of multifilament netting. That's why, before immersing them in seawater, they should be coated with an antifouling agent. It helps to prevent net occlusion and to increase its lifespan. Biofouling in marine aquaculture is a specific problem and has three main neg. effects. It causes net occlusion and so restricts water and oxygen exchange. Besides, the low dissolved oxygen levels from poor water exchange increases the stress levels of fish, lowers immunity and increases vulnerability to disease. Also, the extra wt. imposed by fouling causes cage deformation and structural fatigue. The maintenance and loss of equipment cause the increase of prodn. costs for the industry. Biocides are chem. substances that can prohibit or kill microorganisms responsible for biofouling. The expansion of the aquaculture industry requires the use of more drugs, disinfectants and antifoulant compds. (biocides) to eliminate the microorganisms in the aquaculture facilities. Unfortunately, the use of biocides in the aquatic environment has proved to be harmful as it has toxic effects on the marine environment. The most commonly used biocides in antifouling paints are Tributyltin (TBT), Chlorothalonil, Dichlofluanid, Sea-Nine 211, Diuron, Irgarol 1051 and Zinc Pyrithione. Restrictions were imposed on the use of TBT, that's why org. booster biocides were recently introduced. The replacement products are generally based on copper metal oxides and org. biocides. This paper provides an overview of the effects of antifouling biocides on aquatic organisms. It will focus on the eight booster biocides in common use, despite little data are available for some of them. Toxicity values and effects of these antifoulants will also be mentioned for different species of fish, crustaceans, invertebrates and algae.
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    Borkow, G. Safety of Using Copper Oxide in Medical Devices and Consumer Products. Curr. Chem. Biol. 2012, 6, 86– 92,  DOI: 10.2174/187231312799984349
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    Safety of using copper oxide in medical devices and consumer products
    Borkow, Gadi
    Current Chemical Biology (2012), 6 (1), 86-92CODEN: CCBUB2; ISSN:1872-3136. (Bentham Science Publishers Ltd.)
    A review. Copper has 2 key properties that make it an active ingredient in the medical devices currently being developed. First, copper is an essential trace element needed by humans, which plays a key role in many physiol. processes in different tissues. For example, copper was shown to be involved in angiogenesis and in wound healing. Second, copper has very potent antibacterial, antifungal, antiviral, and acaricidal properties. Recently, a novel technol. was developed that introduces copper oxide particles into polymeric materials, where they serve as a slow release source of copper ions. For example, by this technol., copper oxide contg. wound dressings that enhance wound healing; copper oxide contg. antiviral respiratory masks that reduce the risk of infection; socks that protect from athlete's foot, and acaricidal bedding products that kill dust mites, were developed. While copper oxide is used as the source of copper in mineral and vitamin supplements and is considered safe, its use in medical devices, as well as in industrial and consumer products, is novel. The current manuscript reviews the safety aspects of the use of copper oxide in products that come in contact with open and closed skin. Copper oxide products were tested in 9 clin. trials and in several non-clin. studies and were found to be non-irritating, non-sensitizing, and safe to use, with not even one adverse reaction recorded, both when in contact with intact and broken skin. This is in accordance with the extremely low risk of adverse reactions attributed to dermal exposure to copper.
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    Liu, J.-F.; Min, G.; Ducker, W. A. AFM study of adsorption of cationic surfactants and cationic polyelectrolytes at the silica-water interface. Langmuir 2001, 17, 4895– 4903,  DOI: 10.1021/la0017936
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    AFM study of adsorption of cationic surfactants and cationic polyelectrolytes at the silica-water interface
    Liu, Jun-Fu; Min, Gilbert; Ducker, William A.
    Langmuir (2001), 17 (16), 4895-4903CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
    The adsorption of cationic polyelectrolytes and cationic surfactants from aq. soln. onto silica substrates was examd. using AFM in surface force mode and in surface imaging mode. The polymers were poly(diallyldimethylammonium chloride) (PDADMAC), poly-L-lysine hydrobromide, and polyvinylbenzyltrimethylammonium chloride, and the surfactants were hexadecyltrimethylammonium chloride (CTACl) and hexadecyltrimethylammonium bromide. CTACl forms micelles at the interface between micellar CTACl solns. and silica. These micelles desorbed when the CTACl soln. was replaced with water. An adsorbed layer of CTACl hindered adsorption of PDADMAC. This is because CTACl generates a surface change that has the same sign as the polymer. When PDADMAC adsorbed in the absence of CTACl, it formed a featureless, neutral layer. The PDADMAC did not desorb, even after extended rinsing with water. When an adsorbed layer of PDADMAC was exposed to CTACl soln. above the crit. micelle concn., the AFM image and surface force are very similar to those obsd. when CTACl adsorbs to silica. This adsorbed layer is either spheres or hemispheres.
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    Chan, K. H.; Lai, S. T.; Poon, L. L. M.; Guan, Y.; Yuen, K. Y.; Peiris, J. S. M. Analytical Sensitivity of Rapid Influenza Antigen Detection Tests for Swine-origin Influenza Virus (H1N1). J. Clin. Virol. 2009, 45, 205– 207,  DOI: 10.1016/j.jcv.2009.05.034
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    Analytical sensitivity of rapid influenza antigen detection tests for swine-origin influenza virus (H1N1)
    Chan, K. H.; Lai, S. T.; Poon, L. L. M.; Guan, Y.; Yuen, K. Y.; Peiris, J. S. M.
    Journal of Clinical Virology (2009), 45 (3), 205-207CODEN: JCVIFB; ISSN:1386-6532. (Elsevier B.V.)
    Background: A novel swine origin influenza virus (S-OIV) (H1N1) is spreading worldwide and threatens to become pandemic. Objectives: Det. anal. sensitivity of selected com. available rapid influenza antigen detection tests in detecting S-OIV H1N1. Study design: Serial dilns. of two S-OIV isolates, one seasonal influenza A (H1N1) isolate and a nasopharyngeal aspirate from a patient with S-OIV disease were tested in five com. available influenza antigen detection tests and by virus isolation in cell culture. Viral M gene copy no. was detd. by quant. PCR methods. Results: The anal. sensitivity of the five influenza antigen detection tests for S-OIV (tissue culture infectious dose 50 (TCID50)) log10 3.3-4.7 was comparable with that of seasonal influenza (TCID50 log10 4.0-4.5). Conclusion: The anal. sensitivity of the selected influenza A antigen detection tests for detection of S-IOV was comparable with that of seasonal influenza H1N1.
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    Malenovska, H. Virus Quantitation by Transmission Electron Microscopy, TCID50, and the Role of Timing Virus Harvesting: A Case Study of Three Animal Viruses. J. Virol. Methods 2013, 191, 136– 140,  DOI: 10.1016/j.jviromet.2013.04.008
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    Virus quantitation by transmission electron microscopy, TCID50, and the role of timing virus harvesting: A case study of three animal viruses
    Malenovska, Hana
    Journal of Virological Methods (2013), 191 (2), 136-140CODEN: JVMEDH; ISSN:0166-0934. (Elsevier B.V.)
    Quantitation of viruses is practised widely in both basic and applied virol. Infectious titrn. in cell cultures, the most common approach to it, is quite labour-intensive and alternative protocols are therefore sought. One of the alternatives is transmission electron microscope (TEM) quantitation using latex particles at a known concn. as a ref. for counting virus particles. If virus TCID50 is detd. in parallel, the ratio of infectious to non-infectious virus particles may be established. This study employs such an approach to compute the no. of virus particles and TCID50, and establish their correlation for three viruses: Canine adenovirus 1 (CAdV-1), Feline calicivirus (FCV) and Bovine herpesvirus 1 (BoHV-1). Each of the viruses was grown in five replicates until complete cytopathol. was recorded (time 0), then frozen. They were thawed, filter-sterilised and left for addnl. periods of 16, 32 and 48 h at 37 °C. At each time point, the infectious ability of the virus was characterised by TCID50 and the no. of virions quantified by TEM, in order to evaluate the influence of timing on virus harvest. The virus particle count detd. by TEM did not change for any of the viruses throughout the expt. The relationship between virus particle counts with TCID50 at time 0 showed good linearity response; their ratio was almost const. The virus particle-to-TCID50 ratio varied between 146 and 426 (mean ± SD: 282 ± 103) for CAdV-1, between 36 and 79 (57 ± 18) for FCV and between 110 and 249 (167 ± 53) for BoHV-1. The proportion of non-infectious particles did not change throughout the expt. for either CAdV-1 or BoHV-1. However, a decrease in virus infectious ability disclosed by TCID50 indicated that the fraction of non-infectious particles in FCV increased 300,000 times when time 0 and 48 h were compared. The quantitation of viruses with TEM is a simple and rapid protocol for virus quantitation but account must be taken of the type of virus and harvesting time as virus counts need not necessarily correlate with virus infectious ability.
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    Reed, L. J.; Muench, H. A Simple Method of Estimating Fifty per cent Endpoints. Am. J. Epidemiol. 1938, 27, 493– 497,  DOI: 10.1093/oxfordjournals.aje.a118408
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    Fujimori, Y.; Sato, T.; Hayata, T.; Nagao, T.; Nakayama, M.; Nakayama, T.; Sugamata, R.; Suzuki, K. Novel Antiviral Characteristics of Nanosized Copper (I) Iodide Particles Showing Inactivation Activity against 2009 Pandemic H1N1 Influenza Virus. Appl. Environ. Microbiol. 2012, 78, 951– 955,  DOI: 10.1128/AEM.06284-11
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    Novel antiviral characteristics of nanosized copper(I) iodide particles showing inactivation activity against 2009 pandemic H1N1 influenza virus
    Fujimori, Yoshie; Sato, Tetsuya; Hayata, Taishi; Nagao, Tomokazu; Nakayama, Mikio; Nakayama, Tsuruo; Sugamata, Ryuichi; Suzuki, Kazuo
    Applied and Environmental Microbiology (2012), 78 (4), 951-955CODEN: AEMIDF; ISSN:0099-2240. (American Society for Microbiology)
    We investigated the antiviral activity of nanosized copper(I) iodide (CuI) particles having an av. size of 160 nm. CuI particles showed aq. stability and generated hydroxyl radicals, which were probably derived from monovalent copper (Cu+). We confirmed that CuI particles showed antiviral activity against an influenza A virus of swine origin (pandemic [H1N1] 2009) by plaque titrn. assay. The virus titer decreased in a dose-dependent manner upon incubation with CuI particles, with the 50% effective concn. being approx. 17 μg/mL after exposure for 60 min. SDS-PAGE anal. confirmed the inactivation of the virus due to the degrdn. of viral proteins such as hemagglutinin and neuraminidase by CuI. ESR spectroscopy revealed that CuI generates hydroxyl radicals in aq. soln., and radical prodn. was found to be blocked by the radical scavenger N-acetylcysteine. Taken together, these findings indicate that CuI particles exert antiviral activity by generating hydroxyl radicals. Thus, CuI may be a useful material for protecting against viral attacks and may be suitable for applications such as filters, face masks, protective clothing, and kitchen cloths.
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    Hans, M.; Mathews, S.; Mücklich, F.; Solioz, M. Physicochemical Properties of Copper Important for its Antibacterial Activity and Development of a Unified Model. Biointerphases 2016, 11, 018902  DOI: 10.1116/1.4935853
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    Physicochemical properties of copper important for its antibacterial activity and development of a unified model
    Hans, Michael; Mathews, Salima; Muecklich, Frank; Solioz, Marc
    Biointerphases (2016), 11 (1), 018902/1-018902/8CODEN: BJIOBN; ISSN:1559-4106. (American Institute of Physics)
    Contact killing is a novel term describing the killing of bacteria when they come in contact with metallic copper or copper-contg. alloys. In recent years, the mechanism of contact killing has received much attention and many mechanistic details are available. The authors here review some of these mechanistic aspects with a focus on the crit. physicochem. properties of copper which make it antibacterial. Known mechanisms of contact killing are set in context to ionic, corrosive, and phys. properties of copper. The anal. reveals that the oxidn. behavior of copper, paired with the soly. properties of copper oxides, are the key factors which make metallic copper antibacterial. The concept advanced here explains the unique position of copper as an antibacterial metal. Based on our model, novel design criteria for metallic antibacterial materials may be derived. (c) 2016 American Institute of Physics.
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    Chang, Y.-R.; Taylor, S.; Duncan, S.; Mazilu, D. A.; Ritter, A. L.; Ducker, W. A. Fabrication of Stabilized Colloidal Crystal Monolayers. Colloids Surf., A 2017, 514, 185– 191,  DOI: 10.1016/j.colsurfa.2016.11.050
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    Fabrication of stabilized colloidal crystal monolayers
    Chang, Yow-Ren; Taylor, Shane; Duncan, Scott; Mazilu, Dan A.; Ritter, A. L.; Ducker, William A.
    Colloids and Surfaces, A: Physicochemical and Engineering Aspects (2017), 514 (), 185-191CODEN: CPEAEH; ISSN:0927-7757. (Elsevier B.V.)
    A recent report described a very simple and rapid method to prep. colloidal crystal monolayers by rubbing spherical particles between two rubber plates [Park et al., Advanced Materials, 26 (2014) 4633]. Here we describe a commensurately simple extension of Park's procedure to prep. films that are much more robust, yet retain the overall structure of the colloidal crystal monolayer. The procedure produces solid necks that connect pairs of particles and also connect particles to the solid. These connections between particles are achieved by first transporting liq. to menisci between the particles and then solidifying those necks by exposure to gas phase reactant and catalyst. We show that the stabilized films are much more resistant to removal of particles during a peel test. We also show that the stabilization method is effective on silica layer-by-layer films.
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  3. Florencia A. Monge, Pradeepkumar Jagadesan, Virginie Bondu, Patrick L. Donabedian, Linnea Ista, Eva Y. Chi, Kirk S. Schanze, David G. Whitten, Alison M. Kell. Highly Effective Inactivation of SARS-CoV-2 by Conjugated Polymers and Oligomers. ACS Applied Materials & Interfaces 2020, 12 (50) , 55688-55695. https://doi.org/10.1021/acsami.0c17445
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  • Abstract

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    Figure 1

    Figure 1. Cross-section view of the Cu2O/polyurethane film.

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    Figure 2

    Figure 2. Time course of viable titer of SARS-CoV-2 on solids with and without a coating of cuprous oxide microparticles bound with polyurethane (Cu2O/PU). Note that the vertical axis is on a log10 scale. Data is shown for coated glass and coated stainless steel. Individual circular data points represent each independent measurement, and the × symbol represents the mean of the log of independent measurements. The detection limit was 90 TCID50/mL (shown with a dotted line). Experimental results where the virus was not detected are plotted at 90 TCID50/mL and are included in the average as 90 TCID50/mL. SARS-CoV-2 is inactivated much more rapidly on the coated surface than on the bare surface.

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    Figure 3

    Figure 3. Time course of viable titer of SARS-CoV-2 on glass coated in Cu2O/PU that was subjected to five cycles of exposure to SARS-CoV-2 plus soaking in 70% ethanol. The uncoated glass was also subjected to the disinfection cycles.

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      AFM study of adsorption of cationic surfactants and cationic polyelectrolytes at the silica-water interface
      Liu, Jun-Fu; Min, Gilbert; Ducker, William A.
      Langmuir (2001), 17 (16), 4895-4903CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)
      The adsorption of cationic polyelectrolytes and cationic surfactants from aq. soln. onto silica substrates was examd. using AFM in surface force mode and in surface imaging mode. The polymers were poly(diallyldimethylammonium chloride) (PDADMAC), poly-L-lysine hydrobromide, and polyvinylbenzyltrimethylammonium chloride, and the surfactants were hexadecyltrimethylammonium chloride (CTACl) and hexadecyltrimethylammonium bromide. CTACl forms micelles at the interface between micellar CTACl solns. and silica. These micelles desorbed when the CTACl soln. was replaced with water. An adsorbed layer of CTACl hindered adsorption of PDADMAC. This is because CTACl generates a surface change that has the same sign as the polymer. When PDADMAC adsorbed in the absence of CTACl, it formed a featureless, neutral layer. The PDADMAC did not desorb, even after extended rinsing with water. When an adsorbed layer of PDADMAC was exposed to CTACl soln. above the crit. micelle concn., the AFM image and surface force are very similar to those obsd. when CTACl adsorbs to silica. This adsorbed layer is either spheres or hemispheres.
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      Chan, K. H.; Lai, S. T.; Poon, L. L. M.; Guan, Y.; Yuen, K. Y.; Peiris, J. S. M. Analytical Sensitivity of Rapid Influenza Antigen Detection Tests for Swine-origin Influenza Virus (H1N1). J. Clin. Virol. 2009, 45, 205– 207,  DOI: 10.1016/j.jcv.2009.05.034
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      Analytical sensitivity of rapid influenza antigen detection tests for swine-origin influenza virus (H1N1)
      Chan, K. H.; Lai, S. T.; Poon, L. L. M.; Guan, Y.; Yuen, K. Y.; Peiris, J. S. M.
      Journal of Clinical Virology (2009), 45 (3), 205-207CODEN: JCVIFB; ISSN:1386-6532. (Elsevier B.V.)
      Background: A novel swine origin influenza virus (S-OIV) (H1N1) is spreading worldwide and threatens to become pandemic. Objectives: Det. anal. sensitivity of selected com. available rapid influenza antigen detection tests in detecting S-OIV H1N1. Study design: Serial dilns. of two S-OIV isolates, one seasonal influenza A (H1N1) isolate and a nasopharyngeal aspirate from a patient with S-OIV disease were tested in five com. available influenza antigen detection tests and by virus isolation in cell culture. Viral M gene copy no. was detd. by quant. PCR methods. Results: The anal. sensitivity of the five influenza antigen detection tests for S-OIV (tissue culture infectious dose 50 (TCID50)) log10 3.3-4.7 was comparable with that of seasonal influenza (TCID50 log10 4.0-4.5). Conclusion: The anal. sensitivity of the selected influenza A antigen detection tests for detection of S-IOV was comparable with that of seasonal influenza H1N1.
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      Malenovska, H. Virus Quantitation by Transmission Electron Microscopy, TCID50, and the Role of Timing Virus Harvesting: A Case Study of Three Animal Viruses. J. Virol. Methods 2013, 191, 136– 140,  DOI: 10.1016/j.jviromet.2013.04.008
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      Virus quantitation by transmission electron microscopy, TCID50, and the role of timing virus harvesting: A case study of three animal viruses
      Malenovska, Hana
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      Quantitation of viruses is practised widely in both basic and applied virol. Infectious titrn. in cell cultures, the most common approach to it, is quite labour-intensive and alternative protocols are therefore sought. One of the alternatives is transmission electron microscope (TEM) quantitation using latex particles at a known concn. as a ref. for counting virus particles. If virus TCID50 is detd. in parallel, the ratio of infectious to non-infectious virus particles may be established. This study employs such an approach to compute the no. of virus particles and TCID50, and establish their correlation for three viruses: Canine adenovirus 1 (CAdV-1), Feline calicivirus (FCV) and Bovine herpesvirus 1 (BoHV-1). Each of the viruses was grown in five replicates until complete cytopathol. was recorded (time 0), then frozen. They were thawed, filter-sterilised and left for addnl. periods of 16, 32 and 48 h at 37 °C. At each time point, the infectious ability of the virus was characterised by TCID50 and the no. of virions quantified by TEM, in order to evaluate the influence of timing on virus harvest. The virus particle count detd. by TEM did not change for any of the viruses throughout the expt. The relationship between virus particle counts with TCID50 at time 0 showed good linearity response; their ratio was almost const. The virus particle-to-TCID50 ratio varied between 146 and 426 (mean ± SD: 282 ± 103) for CAdV-1, between 36 and 79 (57 ± 18) for FCV and between 110 and 249 (167 ± 53) for BoHV-1. The proportion of non-infectious particles did not change throughout the expt. for either CAdV-1 or BoHV-1. However, a decrease in virus infectious ability disclosed by TCID50 indicated that the fraction of non-infectious particles in FCV increased 300,000 times when time 0 and 48 h were compared. The quantitation of viruses with TEM is a simple and rapid protocol for virus quantitation but account must be taken of the type of virus and harvesting time as virus counts need not necessarily correlate with virus infectious ability.
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      Fujimori, Y.; Sato, T.; Hayata, T.; Nagao, T.; Nakayama, M.; Nakayama, T.; Sugamata, R.; Suzuki, K. Novel Antiviral Characteristics of Nanosized Copper (I) Iodide Particles Showing Inactivation Activity against 2009 Pandemic H1N1 Influenza Virus. Appl. Environ. Microbiol. 2012, 78, 951– 955,  DOI: 10.1128/AEM.06284-11
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      Novel antiviral characteristics of nanosized copper(I) iodide particles showing inactivation activity against 2009 pandemic H1N1 influenza virus
      Fujimori, Yoshie; Sato, Tetsuya; Hayata, Taishi; Nagao, Tomokazu; Nakayama, Mikio; Nakayama, Tsuruo; Sugamata, Ryuichi; Suzuki, Kazuo
      Applied and Environmental Microbiology (2012), 78 (4), 951-955CODEN: AEMIDF; ISSN:0099-2240. (American Society for Microbiology)
      We investigated the antiviral activity of nanosized copper(I) iodide (CuI) particles having an av. size of 160 nm. CuI particles showed aq. stability and generated hydroxyl radicals, which were probably derived from monovalent copper (Cu+). We confirmed that CuI particles showed antiviral activity against an influenza A virus of swine origin (pandemic [H1N1] 2009) by plaque titrn. assay. The virus titer decreased in a dose-dependent manner upon incubation with CuI particles, with the 50% effective concn. being approx. 17 μg/mL after exposure for 60 min. SDS-PAGE anal. confirmed the inactivation of the virus due to the degrdn. of viral proteins such as hemagglutinin and neuraminidase by CuI. ESR spectroscopy revealed that CuI generates hydroxyl radicals in aq. soln., and radical prodn. was found to be blocked by the radical scavenger N-acetylcysteine. Taken together, these findings indicate that CuI particles exert antiviral activity by generating hydroxyl radicals. Thus, CuI may be a useful material for protecting against viral attacks and may be suitable for applications such as filters, face masks, protective clothing, and kitchen cloths.
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      Hans, M.; Mathews, S.; Mücklich, F.; Solioz, M. Physicochemical Properties of Copper Important for its Antibacterial Activity and Development of a Unified Model. Biointerphases 2016, 11, 018902  DOI: 10.1116/1.4935853
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      Physicochemical properties of copper important for its antibacterial activity and development of a unified model
      Hans, Michael; Mathews, Salima; Muecklich, Frank; Solioz, Marc
      Biointerphases (2016), 11 (1), 018902/1-018902/8CODEN: BJIOBN; ISSN:1559-4106. (American Institute of Physics)
      Contact killing is a novel term describing the killing of bacteria when they come in contact with metallic copper or copper-contg. alloys. In recent years, the mechanism of contact killing has received much attention and many mechanistic details are available. The authors here review some of these mechanistic aspects with a focus on the crit. physicochem. properties of copper which make it antibacterial. Known mechanisms of contact killing are set in context to ionic, corrosive, and phys. properties of copper. The anal. reveals that the oxidn. behavior of copper, paired with the soly. properties of copper oxides, are the key factors which make metallic copper antibacterial. The concept advanced here explains the unique position of copper as an antibacterial metal. Based on our model, novel design criteria for metallic antibacterial materials may be derived. (c) 2016 American Institute of Physics.
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      Chang, Y.-R.; Taylor, S.; Duncan, S.; Mazilu, D. A.; Ritter, A. L.; Ducker, W. A. Fabrication of Stabilized Colloidal Crystal Monolayers. Colloids Surf., A 2017, 514, 185– 191,  DOI: 10.1016/j.colsurfa.2016.11.050
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      Fabrication of stabilized colloidal crystal monolayers
      Chang, Yow-Ren; Taylor, Shane; Duncan, Scott; Mazilu, Dan A.; Ritter, A. L.; Ducker, William A.
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      A recent report described a very simple and rapid method to prep. colloidal crystal monolayers by rubbing spherical particles between two rubber plates [Park et al., Advanced Materials, 26 (2014) 4633]. Here we describe a commensurately simple extension of Park's procedure to prep. films that are much more robust, yet retain the overall structure of the colloidal crystal monolayer. The procedure produces solid necks that connect pairs of particles and also connect particles to the solid. These connections between particles are achieved by first transporting liq. to menisci between the particles and then solidifying those necks by exposure to gas phase reactant and catalyst. We show that the stabilized films are much more resistant to removal of particles during a peel test. We also show that the stabilization method is effective on silica layer-by-layer films.
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    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acsami.0c11425.

    • Chemical composition of Cu2O particles (Table S1), elemental composition of the Cu2O/PU film measured by EDS (Table S2), virus titration data (Tables S3–S10), alternative methods for preparing the Cu2O/ PU film, particle size distribution of Cu2O particles (Figure S1), SEM Images of the Cu2O/PU coating before and after plasma cleaning (Figure S2), XPS results for Cu2O/PU (Figure S3), XPS results for the PA polymer films (Figure S4), XPS results for the PDADMAC polymer films (Figure S5), histogram of residuals for the virus titer (Figure S6), viral titer for the polyurethane film (Figure S7), viral titer for the high-contact angle sample (Figure S8), images of various items coated in Cu2O/PU (Figure S9), viral titer for samples stored under water for 13 days (Figure S10), cross-hatch test results for Cu2O/PU on stainless steel (Figure S11), peel test results (Figure S12), viral titer for PA (polyallylamine) (Figure S13), and viral titer for PDADMAC (Figure S14) (PDF)


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