Log10 Of Vesicular Stomatitis Virus Research Articles

Evaluation of Earth Sense® Neutral Disinfectant Detergent as an Alternative to Micro-Chem Plus™ Detergent Disinfectant for Use in BSL-4 Laboratories using Vesicular Stomatitis Virus as a Surrogate

Micro-Chem Plus (MCP) detergent disinfectant cleaner is a common chemical disinfectant currently used at most biosafety level 4 laboratories to ensure proper disinfection and decontamination of waste contaminated with risk group 4 agents. However, similar to a number of disinfectants, MCP contains surfactants known as nonylphenol ethoxylates that are considered highly toxic to the aquatic environment. An alternate disinfectant, Earth Sense (ES) neutral disinfection detergent, was identified as one that lacks nonylphenol ethoxylates. This study compares the virucidal efficacy profiles of both MCP and ES using Vesicular Stomatitis Virus (VSV) as a surrogate for risk group 4 agents. Results indicate that a 5% solution of ES with a contact time of 5 minutes was comparable to 5% MCP solution and reduced VSV titer by at least 6 log10. In addition, both 5% solutions of ES and MCP disinfectants were stable for at least 5 months and were able to inactivate 6 log10 of VSV. These results demonstrate that ES detergent may be a potential option to replace MCP as a suitable disinfectant against risk group 4 agents due to its comparable virucidal efficacy as well as its greener environmental profile.

Virus photoinactivation in stroma-free hemoglobin with methylene blue or 1,9-dimethylmethylene blue.

Photoinactivation of vesicular stomatitis virus (VSV) in stroma-free hemoglobin (SFH) was carried out using methylene blue (MB) or 1,9-dimethylmethylene blue (DMMB). The VSV was more sensitive to inactivation by 660 nm light with 1 microM DMMB than with the same concentration of MB. Under conditions that inactivated 6 log10 of VSV, the methemoglobin content (Met-Hb[%]) and P50 of hemoglobin were changed by 1 microM MB phototreatment but were not changed by 1 microM DMMB phototreatment. The migration of hemoglobin during electrophoresis and the activity of superoxide dismutase were not changed by MB or DMMB phototreatment. In contrast to the results obtained with DMMB at 660 nm, 580 nm irradiation of SFH with DMMB resulted in a significant increase of Met-Hb(%) under conditions that only inactivated 1.19 log10 VSV. The 580 nm irradiation primarily activates the dimer and higher-order aggregates of the dyes, while 660 nm irradiation primarily activates the monomer. These results indicate that the monomer form of DMMB can effectively inactivate viruses without damage to SFH.

Differential sensitivities of viruses in red cell suspensions to methylene blue photosensitization

Previous studies explored the feasibility of using the photosensitizer methylene blue (MB) as a virucidal agent in red cell suspensions. Under treatment conditions (5 microM [5 mumol/L] MB, 3.4 x 10(4) J/m2) that resulted in more than 6 log10 inactivation of vesicular stomatitis virus (VSV) or of the enveloped bacteriophage phi 6, red cell membrane alterations were observed. Increased red cell ion permeability and the binding of plasma proteins to the red cell surface were the most sensitive indicators, which varied in a dose-dependent fashion. Inactivation of three additional extracellular viruses and intracellular human immunodeficiency virus type 1 (HIV-1) was assessed after MB phototreatment of red cell suspensions. Potassium leakage and IgG binding also were characterized in MB-treated red cell suspensions that were exposed to low-fluence light (6 x 10(3) J/m2). Different viruses exhibit a wide range of sensitivities to MB photoinactivation. For example, phototreatment conditions (5 microM [5 mumol/L] MB, 3.4 x 10(4) J/m2) that inactivated more than 6 log10 of VSV did not inactivate the nonenveloped picornavirus, encephalomyocarditis virus. In contrast, lower fluences (6 x 10(3) J/m2) inactivated approximately 5 log10 or more of Sindbis virus and approximately 4log10 of extracellular HIV-1. These less stringent phototreatment conditions (5 microM [5 mumol/L] 6 x 10(3) J/m2) caused lower rates of red cell potassium leakage (reduction by 6-fold) and little or no binding of plasma proteins to the red cell surface, compared to values observed previously with higher fluences. However, neither 6 x 10(3) nor 4.1 x 10(4) J per m2 fluences resulted in any inactivation of intracellular HIV as represented by changes in the amount of p24 antigen produced during co-culture of actively infected H9 cells. MB-based protocols would require the use of high-efficiency (> 6log10) white cell-reduction filters or additional inactivation steps to deplete or inactivate intracellular virus.

Inactivation of lipid-enveloped and non-lipid-enveloped model viruses in normal human plasma by crosslinked starch-iodine.

The purpose of this study was to evaluate an iodine-based method of activating potentially harmful viruses that might be found in normal human plasma. A procedure has been developed for inactivating lipid-enveloped and non-lipid-enveloped model viruses in normal human plasma by using iodine complexed to crosslinked potato starch. The established conditions are an iodine concentration of 1.05 mg per mL and 60 minutes' incubation. Under these conditions, inactivation of more than 9 log10 of vesicular stomatitis virus, a lipid-enveloped virus, and more than 7 log10 of encephalomyocarditis virus, a non-lipid-enveloped virus, was achieved, with minimal losses of biologic activity of selected plasma proteins. Under these conditions, 70 percent of factor VIII activity, 77 percent of factor IX activity, and 100 percent of protein C activity in the plasma were retained. Crosslinked starch-iodine may be useful in the inactivation of viruses in single-donor plasma units and in pooled human plasma before fractionation.

Virucidal levels of ozone induce hemolysis and hemoglobin degradation

The animal virus, vesicular stomatitis virus (VSV), and the bacterial virus, phi 6, were inactivated by greater than 4 log10 in response to incubation with 13 to 14 mL of 1.4 mmol per L (65 micrograms/mL) to 1.6 mmol per L (75 micrograms/mL) of overlaid ozone in virus-spiked, dilute, red cell suspensions. Virus inactivation was greatly inhibited when ozone was overlaid in the presence of high-hematocrit red cells or, to a lesser degree, high levels of plasma. At hematocrits at which 5 to 6 log10 of VSV were inactivated, ozone caused 30-percent hemolysis, as measured by the loss of total cellular hemoglobin. Unexpectedly, this level of hemolysis could not be observed in supernatants because of the ozone-induced destruction (bleaching) of extracellular hemoglobin. These results suggest that ozone may have little biological specificity for damaging viruses over red cells.

Inactivation of viruses in platelet suspensions that retain their in vitro characteristics: comparison of psoralen‐ultraviolet A and merocyanine 540‐visible light methods

The ability of two fundamentally different photochemical procedures to inactivate model viruses in platelet suspensions was compared. Merocyanine 540 (MC 540) with visible light was used as an example of an oxygen-dependent chemical-directed at the viral membrane, and aminomethyl trimethyl psoralen (AMT) with ultraviolet A light (UVA) was used as an example of a nucleic acid-directed system. Antiviral conditions in petri dishes were identified and the effects of these procedures on platelet suspensions in plastic storage containers were studied. Concentrations of photochemicals in the 10 to 150 mumol range with 30 to 60 minutes of visible light (MC 540) or 1 to 2 minutes of UVA (AMT) readily inactivated 5 to 6 log10 of vesicular stomatitis virus (VSV) and other model viruses in platelet suspensions, provided the plasma concentration was reduced to about 15 percent by the use of a synthetic platelet storage medium. Extracellular pH, morphology scores, and aggregation response dropped markedly when platelets were treated with MC 540 and visible light. However, treatment with 136 mumol per L of AMT and 1 to 3 minutes of UVA could inactivate 5 log10 of VSV in platelet suspensions with retention of platelet characteristics for 4 days, particularly if oxygen levels were reduced during treatment. These studies demonstrate that AMT-UVA treatment meets the initial requirements for virus inactivation in platelet suspensions.

Large-scale production and properties of a solvent-detergent-treated factor IX concentrate from human plasma.

A human solvent-detergent (SD)-treated factor IX concentrate has been produced from cryoprecipitate-poor plasma using DEAE-Sepharose CL-6B and heparin-Sepharose CL-6B chromatography. The DEAE eluate was incubated with an SD mixture [0.3% tri(n-butyl) phosphate-1% Tween 80, 6-h at 24 degrees C] which was found to inactivate, in less than 1 h, more than 3.8 log10 of vesicular stomatitis virus and more than 4.8 log10 of Sindbis virus; the SD was removed by a subsequent heparin adsorption step. The specific activity of the concentrate was 10.9 +/- 1.3 IU factor IX: c/mg protein (n = 15). The factor IX coagulant to antigen ratio was 0.7 +/- 0.1. The concentrate was essentially free of factors II, VII and X, and protein C. The usual major contaminants of prothrombin complex concentrate (PCC) were absent: the concentrate contained about 94% alpha-1 proteins, and only 4 major proteins were resolved by SDS-PAGE (respective apparent molecular weight: 130, 86, 76 and 69 kilodaltons), and by crossed immunoelectrophoresis against an anti-PCC serum. The nonactivated partial thromboplastin time was equivalent to that of PCC; the product was devoid of factor IXa, of other activated procoagulant factors and of coagulant-active phospholipids (removed with SD in the heparin breakthrough fraction). Animal studies using the Wessler test and acute-toxicity test in rabbits revealed no adverse side effects. SD treatment could thus be used to inactivate viruses in factor IX concentrate and improve the safety of replacement therapy in hemophilia B.