Variations in time, temperature, concentration, pH, and relative humidity may affect the sporicidal activity of various agents. Accordingly, and especially for real-world situations, attention must be paid simultaneously to more than one controllable or uncontrollable factor. In Tables 1 and 2 and in the discussion, we address some of the key ancillary factors.

Boiling water for >10 minutes, for example, can reduce B. anthracis spore counts by at least 10⁶ (Table 1). Variations in time and temperature conditions required to reduce spore counts listed in Table 1 can be attributed to differences in experimental conditions, strains of B. anthracis tested, or inoculum size.

The U.S. Environmental Protection Agency indicates that use of sodium hypochlorite as a sporicide is applicable under an emergency exemption (Section 18: Crisis Exemption; Federal Insecticide, Fungicide, and Rodenticide Act); as such, sodium hypochlorite may be used under the conditions specified (32). Given these conditions, the sporicidal effectiveness of hypochlorite solutions depends on the concentration of free available chlorine and pH. Common household bleach (sodium hypochlorite) has a pH of 12 to prolong its shelf life. To achieve effective sporicidal activity, bleach must be diluted with water to increase the free available chlorine and acetic acid to change the pH of the solution to 7 (11). Organic matter may decrease the sporicidal efficiency of sodium hypochlorite (33).

Concentration, humidity, temperature, and carrier material affect gaseous sterilization of spores. Ethylene oxide penetrates into porous material (absorbed strongly by rubber and many plastics); thus vapors are not readily eliminated by brief aeration. Ethylene oxide is also flammable (34). Residual spores were not completely killed after a 30-minute exposure to chlorine dioxide at a relative humidity of 20% to 40%, whereas all spores were killed after a 15-minute exposure to chlorine dioxide with the addition of prehumidification at a relative humidity of 70% to 75% (21). Peracetic acid vapor does not penetrate well into porous surfaces and is flammable. The amount of contamination, level of cleanliness of surfaces, and relative humidity will contribute to peracetic acid vapor’s effectiveness as a sporicide (24). Organic matter may absorb and chemically react with propylene oxide, reducing its effectiveness. Organic matter may also provide physical protection from the oxide (25). The sporicidal property of ozone is affected by relative humidity: as relative humidity decreases, the time required for killing organisms increases (27).

Decontamination of buildings from intentional release of B. anthracis is a new problem, and no accumulated scientific knowledge exists on the subject. Two areas of prior scientific research may be relevant: food processing and laboratory decontamination. With modification based on further study, the technologies used in laboratories and food processing plants may be applied to buildings.

Direct information on killing B. anthracis spores in foods by cooking is scarce, and the complexity of food matrices precludes easy extrapolation of the laboratory data into nonfood matrices. However, information on inactivating spores of bacterial species more resistant to environmental conditions than B. anthracis can provide guidance. The spores of Clostridium botulinum are more resistant to heat inactivation than are B. anthracis spores (4). The commercial retort process of canning achieves a 12-log reduction of C. botulinum spores, and by extension, should achieve a similar killing rate for B. anthracis spores. Further research in this area is needed.

Historically, formaldehyde solution or gas has been used both as a disinfectant and chemical sterilant. Formaldehyde was used to disinfect as early as the late 1880s and is still used to reprocess hemodialyzers for reuse on the same patient and to decontaminate biologic safety cabinets and laboratories (35–37). Formaldehyde gas has been used for fumigation in the poultry industry and for disinfection of biologic safety cabinets and laboratories (38,39). Data from controlled experiments with B. globigii NCTC 10073 spores have demonstrated the effect of humidity on formaldehyde concentration (mg/m³) to obtain a >8-log reduction in viable spores (15).

Fumigation with formaldehyde vapor (18 mg/L–21 mg/L) has also been used to treat a textile mill contaminated with ­B. anthracis spores (40). In this instance, contamination was greatly reduced immediately after treatment and was undetectable 6 months later. However, the possible role of formaldehyde as a carcinogen has limited its use. Formaldehyde can be neutralized with ammonium bicarbonate after fumigation, reducing its carcinogenic properties.

Gamma radiation was used in the 1960s and 1970s to disinfect B. anthracis–contaminated imported bailed goat hair. A study by Horne et al. suggested that a dose of 1.5 megarads from a 200,000-rad/hour cobalt source was sufficient to kill most resistant spores when mixed with goat hair; 2 megarads was recommended to include a margin of safety (31). After the intentional release of B. anthracis through the postal system in 2001, pursuing a decontamination method for the undelivered mail was essential. Gamma radiation was used to decontaminate all mail from contaminated facilities on the basis of these data.

Multiple technologies may be needed to decontaminate buildings and their contents. As in a laboratory, where some items are wiped, some items are autoclaved, and some spaces are treated with gas, more than one method may be required for decontamination. Also, for certain decontamination tasks, e.g., cleaning small heat-proof and water-proof objects, more than one option will be available. Further, even within the context of one type of application (e.g., walls; ducts for heating, ventilating, air conditioning, and refrigeration; carpet; and small objects), potentially conflicting priorities exist between bioefficacy, logistics, and safety.

Our review suggests two conclusions. First, additional scientific research is needed. Although transferring the methods used to decontaminate or sterilize laboratory or food industry settings to decontaminating buildings may be useful, this transfer of methods has not been scientifically tested. Also, much of the data available is based on other Bacillus species; more testing with or correlation to B. anthracis contamination is suggested. Second, choosing between technologies is a complex issue, and a formal decision process would be useful. Various parties in the public and private sector have suggested numerous, sometimes disparate, methods for the inactivation of B. anthracis spores in contaminated environments. Further research is needed regarding improved methods for remediation of environments contaminated with B. anthracis spores, and the literature summarized here provides a basis for that effort.