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A quantitative, reproducible method was developed for the evaluation of the virucidal activity of test gases. Using this method, we determined the virucidal effectiveness of a Steri-Vac ethylene oxide gas sterilizer. Wool gabardine material was exposed to high concentrations of herpes simplex, vaccinia, parainfluenza, or polio viruses and was processed through the sterilizer. Two time-temperature cycles of the machine, 29 C for 180 min and 60 C for 48 min, were used in separate experiments. The viruses were exposed to the gas when freshly pipetted onto the fabric or when pipetted on the material and allowed to dry 16 to 24 hr. In two experiments carried out under each condition, the virus titers were reduced by the sterilization process to less than detectable limits. These titer reductions were for the herpes virus ≥ 2.7 to 5.0 log, for vaccinia virus ≥ 4.0 to 6.1 log, for parainfluenza virus ≥ 1.8 to 4.9 log, and for poliovirus ≥ 4.9 to 7.7 log. The observed reductions in virus titers were the same whether the virus-contaminated fabrics were sealed in polyethylene packages or held in open petri dishes during exposure to ethylene oxide.

The persistence of vaccinia virus on wool (blanket and gabardine) and cotton (sheeting, terry cloth, and knit jersey) fabrics was studied. The fabrics were exposed to the virus by three methods: direct contact, aerosol, and virus-containing dust having a high content of textile fibers. Fabrics exposed to virus by each method were held in 35 and 78% relative humidities at 25 C. Virus was recovered for up to 14 weeks from wool fabrics exposed to virus and held in the low humidity. In contrast, virus persisted for shorter periods of time on the cotton fabrics. No virus was detected on terry cloth as early as 3 days after exposure to virus. The virus appeared to be less stable in the high humidity, and the method of exposure of the fabrics to virus apparently had an effect upon the persistence of the agent. On all fabrics, viral persistence was of sufficient duration to be of epidemiological significance.

Eight compounds were tested in vitro for virucidal and antiviral activity against poliovirus and vaccinia virus. These compounds included five quaternary ammonium salts, two bromosalicylanilides, and neomycin sulfate, an antibiotic. None of the compounds was active against poliovirus, but virucidal activity was demonstrated against vaccinia virus with three of the quarternary ammonium compounds: n -alkyl (C14, C12, C16) dimethyl benzyl ammonium chloride, di-isobutyl cresoxy ethoxy ethyl dimethyl benzyl ammonium chloride monohydrate, and n -alkyl (60% C14, 30% C16, 5% C12, 5% C18) dimethyl benzyl ammonium chlorides plus n -alkyl (50% C12, 30% C14, 17% C16, 3% C18) dimethyl ethylbenzyl ammonium chlorides. Wool blanketing, wool gabardine, and cotton sheeting materials were impregnated with the first of the above virucidal compounds, and the persistence of vaccinia virus on these fabrics was compared with the persistence of the agent on nonimpregnated fabrics of the same type held at 25 C in 35 and 78% relative humidity. No virus could be recovered from the chemically treated fabrics at any time after virus exposure, whereas the virus persisted as long as 4 weeks on nonimpregnated materials. Viable vaccinia virus was also found to persist less than 1 day on a cotton fabric finished with a wash-and-wear modified triazone resin. Poliovirus persisted less than 5 days on this wash-and-wear fabric.

The in vivo anti-influenza virus and antivaccinia virus activity of 156 biologically active compounds was determined. One of two criteria was used for evaluating activity against the influenza virus. The criteria were increase in survivor number and mean survival time, and reduction in virus-induced lung consolidation in treated, infected Swiss mice. Increase in survivor number and mean survival time were the criteria for evaluation of antivaccinia virus activity. Several drug doses were tested against two virus concentrations to demonstrate antiviral activity more clearly. Two compounds were considered significantly active against the influenza virus: DL-noformicin (NSC 72942) and amantadine hydrochloride (NSC 83653). Eleven compounds had reproducible activity against vaccinia virus: isatin-β-thiosemicarbazone (NSC 721), 6-azauracil (NSC 3425), 9-α-fluoro-2α-methylhydrocortisone 21-acetate (NSC 12601), 5-[bis(2-chloroethyl)amino]uracil (NSC 34462), 5-iodo-2′-deoxyuridine (NSC 39661), streptonigrin (NSC 45383), N -methylisatin β-thiosemicarbazone (NSC 69811), cytovirin (NSC 91770), 9-β-D-arabinofuranosyladenine (NSC 404241), and 5-(mercaptomethyl)uracil (NSC 529351).

Glutaraldehyde-tanned woolskin pads which are used for the prevention of decubitus ulcers in bed patients were experimentally contaminated with polio or vaccinia viruses. Two methods of exposure, direct contact and aerosol, were used in separate experiments. Attempts were made to remove or inactivate these virus contaminants by laundering the woolskins in a quaternary ammonium disinfectant, a phenolic disinfectant, or alkalinized glutaraldehyde, in combination with an anionic detergent or a nonionic detergent. The effect of a commercial detergent-sanitizer was also studied. The virus titers were significantly reduced in all experiments, but only laundering in glutaraldehyde in combination with either detergent lowered the vaccinia virus titers to below detectable limits. High concentrations of glutaraldehyde altered the texture of the wool and leather apparently by precipitating a component of the detergent onto the fibers. In all the poliovirus experiments, the virus was still detectable on either or both the wool and the leather of the pads after laundering. The rinse water from each experiment was tested for the presence of virus. No vaccinia virus was recovered, but poliovirus was demonstrated in titers up to 10 3 cell culture 50% infectious doses.

The effects of laundering with both anionic and nonionic detergents in cold, warm, and hot water on poliovirus-contaminated cotton sheeting, cotton terry cloth, washable wool shirting, wool blanketing, dull nylon jersey, and dacron/cotton shirting were determined. The fabrics were exposed to virus by aerosolization and direct contact (pipette) in separate studies. Although the results varied with each factor used in the study, virus titers on all the fabrics were generally reduced considerably by the laundering process. When the fabrics were dried for 20 hr after laundering, an additional decline in virus titers was seen, often to below detectable levels. The type of detergent used made little difference in effect on virus titer reduction, but the hot wash water markedly reduced the detectable virus. Fabric type was not a major factor in the majority of the experiments, although virus tended to be eliminated more readily from the nylon jersey, and in warm water the virus persisted longer on wool blanketing material laundered in anionic detergent. Sterile fabrics of each type laundered with similar fabrics which contained virus often became contaminated by the virus during the laundering process. Virus titers ranging from undetectable to 10 3.9 cell culture 50% infectious doses/ml were obtained from samples of the rinse water after warm- and cold-water laundering.

A prolongation in the lives of Swiss mice inoculated intracerebrally with lymphocytic choriomeningitis virus (LCM) was observed after treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). A variety of treatment schedules, including therapy once or twice daily up to 17 days and single treatments at various times after virus inoculation, were employed. Virus titers ranging to greater than 10 4 were detected in the blood and brains of surviving drug-treated animals. In three comparative studies in which different treatment schedules were used, BCNU was shown to exert a protective effect approximately equal to that of methotrexate in LCM virus-infected mice. Tests were also carried out to investigate the activity of BCNU in mice experimentally infected with eastern equine encephalomyelitis (EEE) virus, western equine encephalomyelitis virus, Semliki Forest (SF) virus, herpes simplex virus, influenza virus strain PR8, vaccinia virus strain WR, Rous sarcoma virus, Friend leukemia virus (FLV), and poliovirus. Slight increases in life span were observed in the treated EEE, SF, and influenza PR8 virus-infected animals. Significant reduction in splenomegaly in FLV-infected animals treated with BCNU was demonstrated. The possible mechanisms of LCM virus inhibition by BCNU, on the basis of these and other studies, were postulated to be either specific antiviral activity or inhibition of “lethal” immune response to the LCM virus. Each of these postulates is discussed.