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Activity of Three Disinfectants and Acidified Nitrite Against Clostridium difficile Spores

  • Marlene Wullt (a1), Inga Odenholt (a1) and Mats Walder (a1)



To identify environmentally safe, rapidly acting agents for killing spores of Clostridium difficile in the hospital environment.


Three classic disinfectants (2% glutaraldehyde, 1.6% peracetyl ions, and 70% isopropanol) and acidified nitrite were compared for activity against C. difficile spores. Four strains of C. difficile belonging to different serogroups were tested using a dilution–neutralization method according to preliminary European Standard prEN 14347. For peracetyl ions and acidified nitrite, the subjective cleaning effect and the sporicidal activity was also tested in the presence of organic load.


Peracetyl ions were highly sporicidal and yielded a minimum 4 log10 reduction of germinating spores already at short exposure times, independent of organic load conditions. Isopropanol 70% showed low or no inactivation at all exposure times, whereas glutaraldehyde and acidified nitrite each resulted in an increasing inactivation factor (IF) over time, from an IF greater than 1.4 at 5 minutes of exposure time to greater than 4.1 at 30 minutes. Soiling conditions did not influence the effect of acidified nitrite. There was no difference in the IF among the 4 strains tested for any of the investigated agents. Acidified nitrite demonstrated a good subjective cleaning effect and peracetyl ions demonstrated a satisfactory effect.


Cidal activity was shown against C. difficile spores by glutaraldehyde, peracetyl ions, and acidified nitrite. As acidified nitrite and peracetyl ions are considered to be environmentally safe chemicals, these agents seem well suited for the disinfection of C. difficile spores in the hospital environment.


Corresponding author

Department of Infectious Diseases, Malmö University Hospital, SE- 205 02 Malmö, Sweden


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1.McFarland, LV, Surawicz, CM, Stamm, WE. Risk factors for Clostridium difficile carriage and C. difficile-associated diarrhea in a cohort of hospitalized patients. J Infect Dis 1990;162:678684.
2.Fekety, R, Kim, KH, Brown, D, Batts, DH, Cudmore, M, Silva, J Jr. Epidemiology of antibiotic-associated colitis: isolation of Clostridium difficile from the hospital environment. Am J Med 1981;70:906908.
3.Kaatz, GW, Gitlin, SD, Schaberg, DR, et al.Acquisition of Clostridium difficile from the hospital environment. Am J Epidemiol 1988;127:12891294.
4.Samore, MH, Venkataraman, L, DeGirolami, PC, Arbeit, RD, Karchmer, AW. Clinical and molecular epidemiology of sporadic and clustered cases of nosocomial Clostridium difficile diarrhea. Am J Med 1996;100:3240.
5.Clabots, CR, Johnson, S, Olson, MM, Peterson, LR, Gerding, DN. Acquisition of Clostridium difficile by hospitalized patients: evidence for colonized new admissions as a source of infection. J Infect Dis 1992;166:561567.
6.McFarland, LV, Mulligan, ME, Kwok, RYY, Stamm, WE. Nosocomial acquisition of Clostridium difficile infection. N Engl J Med 1989;320:204210.
7.Wullt, M, Laurell, MH. Low prevalence of nosocomial Clostridium difficile transmission, as determined by comparison of arbitrarily primed PCR and epidemiological data. J Hosp Infect 1999;43:265273.
8.Rutala, WA, Gergen, MF, Weber, DJ. Inactivation of Clostridium difficile spores by disinfectants. Infect Control Hosp Epidemiol 1993;14:3639.
9.Rutala, WA, Gergen, MF, Weber, DJ. Sporicidal activity of chemical ster-ilants used in hospitals. Infect Control Hosp Epidemiol 1993;14:713718.
10.Worsley, MA. Infection control and prevention of Clostridium difficile infection. J Antimicrob Chemother 1998;41(suppl C):5966.
11.Global Consensus Conference. Final recommendations. Am J Infect Control 1999;27:503513.
12.Wilcox, MH, Fawley, WN. Hospital disinfectants and spore formation by Clostridium difficile. Lancet 2000;356:1324.
13.European Committee for Standardization. Chemical Disinfectants: Basic Sporicidal Activity—Test Method and Requirements (Phase 1). DRAFT prEN 14347. Brussels: European Committee for Standardization; 2001.
14.Walder, M, Myrbäck, K-E, Nilsson, B. A method to evaluate the cleaning and disinfectant action of surface disinfectants. J Hosp Infect 1989;13:149159.
15.Mayfield, JL, Leet, T, Miller, J, Mundy, LM. Environmental control to reduce transmission of Clostridium difficile. Clin Infect Dis 2000;31:9951000.
16.Babb, JR, Bradley, CR, Ayliffe, GAJ. Sporicidal activity of glutaraldehy-des and hypochlorites and other factors influencing their selection for the treatment of medical equipment. J Hosp Infect 1980;1:6375.
17.Rutala, WA, Clontz, EP, Weber, DJ, Hoffmann, KK. Disinfection practices for endoscopes and other semicritical items. Infect Control Hosp Epidemiol 1991;12:282288.
18.Benjamin, N, O'Driscoll, F, Dougall, H, et al.Stomach NO synthesis. Nature 1994;368:502.
19.Shank, JL, Silliker, JH, Harper, RH. The effect of nitric oxide on bacteria. Appl Microbiol 1962;10:185189.
20.De Groote, MA, Fang, FC. NO inhibitions: antimicrobial properties of nitric oxide. Clin Infect Dis 1995;21(suppl 2):S162S165.
21.O'Leary, V, Solberg, M. Effect of sodium nitrite inhibition on intracellular thiol groups and on the activity of certain glycolytic enzymes in Clostridium perfringens. Appl Environ Microbiol 1976;31:208212.
22.Klebanoff, SJ. Reactive nitrogen intermediates and antimicrobial activity: role of nitrite. Free Radie Biol Med 1993;14:351360.
23.Rutala, WA, Weber, DJ. Infection control: the role of disinfection and sterilization. J Hosp Infect 1999;43(suppl):S43S55.
24.Dyas, A, Das, BC. The activity of glutaraldehyde against Clostridium difficile. J Hosp Infect 1985;6:4145.
25.Russell, AD. Glutaraldehyde: current status and uses. Infect Control Hosp Epidemiol 1994;15:724733.
26.Coates, D. Sporicidal activity of sodium dichloroisocyanurate, peroxy-gen and glutaraldehyde disinfectants against Bacillus subtilis. J Hosp Infect 1996;32:283294.
27.Fraise, AP. Choosing disinfectants. J Hosp Infect 1999;43:255264.
28.Cowan, RE, Manning, AP, Ayliffe, GA, et al.Aldehyde disinfectants and health in endoscopy units. Gut 1993;34:16411645.
29.Delmée, M, Bulliard, G, Simon, G. Application of a technique for serogrouping Clostridium difficile in an outbreak of antibiotic-associated diarrhoea. J Infect 1986;13:59.
30.Toma, S, Lesiak, G, Magus, M, Lo, H-L, Delmée, M. Serotyping of Clostridium difficile. J Clin Microbiol 1988;26:426428.
31. van Dijck, P, Avesani, V, Delmée, M. Genotyping of outbreak-related and sporadic isolates of Clostridium difficile belonging to serogroup C. J Clin Microbiol 1996;34:30493055.


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