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EBOV inactivation with DF200 Introduction. VIRUS and EBOLA VIRUS (EBOV) Ebola virus (EBOV) is the virus responsible for Ebola virus disease, a type of hemorrhagic fever, a highly contagious and very severe infectious disease that affects both animals and humans. The virus is transmitted by direct contact with infected body fluids such as blood, saliva, sweat, urine or vomit, the cause of the index case (primary case) is unknown. The incubation period varies from 2 to 21 days (often is between 5 -­‐ 12 day). The disease kills up to 90% of people who are infected. A virus is a small infectious agent that replicates only inside the living cells of other organisms. Viruses can infect all types of life forms. The genetic material within virus particles, and the method by which the material is replicated, varies considerably between different types of viruses: DNA viruses (the genome replication of most DNA viruses takes place in the cell's nucleus), RNA viruses (replication usually takes place in the cytoplasm) and Reverse transcribing viruses (reverse transcribing viruses with RNA genomes –retroviruses-­‐, use a DNA intermediate to replicate, whereas those with DNA genomes –pararetroviruses-­‐ use an RNA intermediate during genome replication). Baltimore classification is a classification system that places viruses into one of seven groups depending on a combination of their nucleic acid (DNA or RNA), strandedness (single-­‐stranded or double-­‐stranded), Sense, and method of replication. Group I dsDNA viruses Viruses possess double-­‐stranded DNA. Group II ssDNA viruses. Viruses possess single-­‐stranded DNA. Group III dsRNA viruses Group IV (+)ssRNA viruses Viruses possess positive-­‐sense single-­‐stranded RNA genomes. Group V: (−)ssRNA viruses Viruses possess negative-­‐sense single-­‐stranded RNA genomes. Group VI ssRNA-­‐RT viruses Group VII dsDNA-­‐RT viruses Viruses possess double-­‐stranded RNA genomes. These genomes are always segmented. Viruses possess single-­‐stranded RNA viruses that replicate through a DNA intermediate. Viruses possess double-­‐stranded DNA genomes and replicate using reverse transcriptase. Viruses differ in their susceptibility and resistance to disinfectant and their ability to be inactivated based predominantly on structural and size differences. According to the Klein and Deforest classification for viral groups, the least resistant to disinfectants are the enveloped viruses (ie. influenza, coronavirus, ebola), those with moderate resistance are the large non-­‐
enveloped viruses (ie. adenovirus) and those with most resistance are the small non-­‐enveloped viruses (ie. piconavirus and parvovirus). Advanced Decon Technologies Jan-­‐15 External Communication Ernesto Asensio Sosa R&D Manager Page ~ 1 ~ EBOV inactivation with DF200 Noll and Youngner classified the viruses in three groups: category A Viruses that have lipoprotein envelopes. Category A viruses are the most susceptible Herpesviridae, Paramyxoviridae,Orthomyxoviridae to disinfectants. category B Small (20-­‐30nm) non-­‐enveloped (non-­‐lipid) viruses. Category B viruses are the least susceptible Picornaviridae, Parvoviridae to disinfectants. category C Large non-­‐enveloped (non-­‐lipid) viruses. Category C viruses are intermediately susceptible to disinfectants. (*) susceptibility is referring to the ability of the disinfectant to disrupt surface properties typically important in attachment to cellular receptors, and does not always result in complete degradation and loss of replication functionality of the nucleic acid. Disinfectants can act on microorganisms in two different ways: growth inhibition (bacteriostasis, fungistasis) or lethal action (bactericidal, fungicidal or virucidal effects). Disinfectant product can be considered as a completely virucidal when it produces degradation of capsid proteins and genetic material. EBOLAVIRUS is an enveloped, linear non-­‐segmented, negative-­‐sense single-­‐stranded RNA virus. For genetic material, Ebola virus is classified as (-­‐)ssRNA virus (Group V). Order Mononegavirales, Family Filoviridae, Genus Ebolavirus. Genus Ebolaviruscomprises 5 distinct species: Bundibugyo ebolavirus (BDBV), Zaire ebolavirus (EBOV), Reston ebolavirus (RESTV) Sudan ebolavirus (SUDV) and Taï Forest ebolavirus (TAFV). As an enveloped virus, Ebola virus is classified in category A. Advanced Decon Technologies Jan-­‐15 External Communication Ernesto Asensio Sosa R&D Manager Page ~ 2 ~ EBOV inactivation with DF200 Introduction. DF200. A non-­‐toxic, non-­‐corrosive, biodegradable aqueous based decontaminant formulation (DF) with enhanced physical stability for the rapid mitigation and decontamination of chemical and biological warfare (CBW) agents and toxic hazardous materials has been developed at Sandia National Laboratories. The formulation allows decontamination of areas populated with both people and sensitive equipment; works on all currently anticipated material surfaces and can be incorporated into a wide variety of carriers (foams, gels, fogs, aerosols) that satisfy a wide variety of operational objectives. The basic chemistry of the formula is a combination of two basic active ingredients; quaternary amines and hydrogen peroxide. The formula also contains several stabilizing compounds, which help to extend the shelf life of the formula and control the chemical reaction so that the decontamination occurs in a safe manner. Additional tests indicate that the formulation is effective as a decontaminant on a several variety hazardous bacteria, viruses and materials such as hydrocarbon based compounds as well. Formulation mechanisms of action: •
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Physically denatures biological agent via a surfactant boring holes in their protein armor Oxidizing agents attack the genetic material (DNA). Hydrolyzing agents attack vital biological agent contents and functions Advanced Decon Technologies manufactures and commercializes Sandia National Laboratories’ Decon Formula under Patent License No. 13-­‐C02621. EPA (US Environmental Protection Agency) registered product as Antibacterial, Cleaner, Mildewstat, Decontaminant, Disinfectant, Deodorizer, Fungicide, Algaecide, and Virucide (Including Norovirus and H1N1). DF-­‐200: VIRUCIDAL ACTIVITY. The disinfection mechanism of DF against viruses is most likely a result of the dual synergy between the surfactant oxidizing properties, in addition to these chemistries, peracetate is also produced and further acts as a strong oxidizer. Rapid degradation of capsid proteins and viral RNA has been observed in a relatively short contact time. Surfactant or quaternary ammonium compounds (QAC) are compounds comprised of hydrophilic and lipophilic portion. Mechanisms of action on viral systems include a primary effect on the lipid enveloped with subsequent degradation of viral capsid proteins, protein denaturation and dissociation of enzymes and interaction with lipids. These mechanisms impact on both enveloped and non-­‐enveloped viruses. Advanced Decon Technologies Jan-­‐15 External Communication Ernesto Asensio Sosa R&D Manager Page ~ 3 ~ EBOV inactivation with DF200 Hydrogen peroxide generates free hydroxyl radicals that can break DNA and RNA structures, attack membrane lipids, and produce disruption in virus capsid structure and other essential cellular components. DF-­‐200 has been demonstrated to provide disinfection efficacy against the following viruses: -­‐ Influenza A. -­‐ Influenza B. -­‐ Norovirus (Feline Calcivirus Surrogate). -­‐ Bovine Coranovirus (BVC), Severe Acute Respiratory Syndrome (SARS) surrogate. -­‐ Bovine Enterovirus (BEC). -­‐ Foot-­‐and-­‐mouth disease virus (FMDV). Conclusion. DF200 is virucidal against Influenza A (including H1N1) and Influenza B virus with relatively short contact time. Influenza is a (-­‐)ssRNA virus (Group V), Family Orthomyxoviridae. Influenza and EBOLA are two viruses of the same group (V) with similar virion structure: capsid enveloped (category A) and Helical Capsid Symetry, with similar method of replication. The formulation demonstrated complete inactivation of Influenza A, Influenza B and Norovirus following a 10 minutes exposure time, BCV viral agents were completely inactivated in less than 3 minutes. The results obtained in Influenza A and B and other virus studies and laboratories tests can be applied broadly to other similar viruses like EBOLA with regard to virucidal effectiveness, with a simply conclusion: DF-­‐200 will inactive Ebola virus in a relatively short contact time and high effectiveness. References. 1.
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EPA (US Environmental Protection Agency) Registration: #80346-­‐1/2 Rapid inactivation of SARS-­‐like Coronaviruses. SAND2004-­‐1120 -­‐ SANDIA REPORT. Inactivation of Various Influenza Strains to Model Avian Influenza (Bird Flu) With Various Disinfectant Chemistries. SANDIA REPORT -­‐ SAND2005-­‐7633. Agricultural Pathogen Decontamination Technology. Reducing the treat of infectious agent spread. SAND2006-­‐0182. SANDIA REPORT. (October 2005). Final Study Report. ATS LABS. Virucidal Efficacy of Disinfectant for Use on Inanimate Environmental Surfaces. Virus: Influenza A Virus. (January 24, 2006). Final Study Report. ATS LABS. Virucidal Efficacy of Disinfectant for Use on Inanimate Environmental Surfaces. Virus: Influenza B Virus. (January 24, 2006). Chemical inactivation of Viruses. J.M.Bieker. Kansas State University (2006). Advanced Decon Technologies Jan-­‐15 External Communication Ernesto Asensio Sosa R&D Manager Page ~ 4 ~