Evaluation of Pressurized Steam Disinfection of Surfaces Contaminated by Mycobacterium terrae: A Surrogate for Mycobacterium bovis.

Introduction: Decontamination of farms affected by bovine tuberculosis could be very challenging during outbreaks occurring in the winter with freezing temperatures. Steam treatment has been of practical interest, but information is needed on whether such treatment is able to inactivate the causative agent, Mycobacterium bovis. This study was to evaluate the use of pressurized steam for inactivation of Mycobacterium terrae, a surrogate for M. bovis on various surfaces. Methods: Carrier disks made of steel, wood, or rubber were inoculated with 6.32 ± 0.38 log10 M. terrae. While being held at background temperatures of -20°C, 4°C, or 21°C, these carrier disks were treated with pressurized steam (120°C ± 5°C) for 5, 10, 15, or 20 s. Reduction in colony forming units of M. terrae and temperatures on the top and bottom surfaces of the disks were determined. Results: Complete inactivation of 6 log10 M. terrae on steel and wood disks was achieved by 10 s of steam treatment at all three background temperatures. In comparison, 20 s of steam treatment was needed for the complete inactivation of mycobacteria on rubber disks. Corresponding to the longer treatment time required for mycobacterial inactivation, temperatures on the bottom surface of the rubber disks rose substantially slower than those of the steel and wood disks at all three background temperatures. Conclusion: The results suggested that treatment with pressurized steam has potential for efficient and effective disinfection of surfaces contaminated by mycobacteria at or below freezing temperatures in winter.

Decontamination of Bacillus anthracis Spores at Subzero Temperatures by Complete Submersion.

Introduction: Bacillus anthracis, the etiological agent of anthrax, produces long-lived spores, which are resistant to heat, cold, pH, desiccation, and chemical agents. The spores maintain their ability to produce viable bacteria even after decades, and when inhaled can cause fatal disease in over half of the clinical cases. Owing to these characteristics, anthrax has been repeatedly selected for both bioweapon and bioterrorism use. In the event of a bioterrorism attack, surfaces in the vicinity of the attack will be contaminated, and recovering from such an event requires rapid and effective decontamination. Previous decontamination method development has focused mainly on temperatures >0°C, and have shown poor efficacy at subzero temperatures. Methods: In this study, we demonstrate the use of calcium chloride (CaCl2) as a freezing point depression agent for pH-adjusted sodium hypochlorite (NaOCl) for the effective and rapid decontamination of B. anthracis Sterne strain spores at subzero temperatures. Results: We show the complete decontamination of 106 B. anthracis Sterne strain spores at temperatures as low as -20°C within 2.5 min by submersion in solution containing 25% (w/v) CaCl2, 0.50% NaOCl, and 0.40% (v/v) acetic acid. We also demonstrate significant reduction in number of spores at -28°C. Conclusions: The results show promise for rapidly decontaminating equipment and materials used in the response to bioterrorism events using readily available consumer chemicals. Future study should examine the efficacy of these results on complex surfaces.

Evaluation of the Use of Sea Water as a Diluent for an Accelerated Hydrogen Peroxide Disinfectant for Inactivation of Avian Influenza Virus: A Surrogate for Infectious Salmon Anemia Virus.

Introduction: Use of sea water as a diluent for disinfectants has been of practical interest for control of aquaculture disease outbreaks in sea where fresh water is limited. This study evaluated the use of natural sea water (NSW), artificial sea water (ASW), or standard hard water (SHW) as a diluent for preparation of accelerated hydrogen peroxide (AHP) solutions against an avian influenza virus, a surrogate for the infectious salmon anemia virus. Methods: AHP solutions containing 0.18%, 0.35%, or 0.44% (w/w) of hydrogen peroxide (H2O2), corresponding to 1/40, 1/20, and 1/16 dilutions of the disinfectant concentrate, were evaluated at -20°C, 4°C, and 21°C. Results: When NSW was used as the diluent, a 0.35% H2O2 concentration was required to inactivate ∼6 log10 virus at 21°C in a 5-min contact time. When temperature dropped to 4°C, 0.44% H2O2 in NSW was required to obtain a similar inactivation within a 5-min contact time. At -20°C, supplemented with antifreeze agents, the 0.44% H2O2 in NSW solutions produced complete inactivation of 5.4 log10 virus within a 10-min contact time. In comparison, lower H2O2 concentrations and/or shorter contact times were needed to inactivate equal amounts of the virus at the same temperature when using SHW or ASW as a diluent to prepare disinfection solutions. Conclusion: The results suggested that NSW could be used as a diluent in disinfection solutions for virus inactivation as long as disinfectant concentrations and/or contact times are properly increased.

Enhanced inactivation of avian influenza virus at -20°C by disinfectants supplemented with calcium chloride or other antifreeze agents.

Avian influenza outbreaks have occurred during winter months, and effective disinfection of poultry premises at freezing temperatures is needed. The commercial disinfectants Virkon and Accel, supplemented with an antifreeze agent [propylene glycol (PG), methanol (MeOH), or calcium chloride (CaCl2)], were evaluated for their effectiveness in killing avian influenza virus (AIV) at -20°C or 21°C. An AIV suspension was applied to stainless steel disks, air-dried, and covered with a disinfectant or antifreeze agent for 5 to 30 min. Virkon (2%) and Accel (6.25%) with 30% PG, 20% MeOH, or 20% CaCl2 inactivated 6 log10 AIV within 5 min at -20°C and 21°C. At these temperatures PG and MeOH alone did not kill AIV, but the 20% CaCl2 solution alone inactivated 5 log10 AIV within 10 min. The results suggested that CaCl2 is potentially useful to enhance the effectiveness of disinfection of poultry facilities after outbreaks of AIV infection in warm and cold seasons.

Les poussées de cas d'influenza aviaire se sont produites durant les mois d'hiver, et une désinfection efficace des sites d'élevage à des températures de congélation est nécessaire. Les désinfectants commerciaux Virkon et Accel, auxquels on ajouta un antigel [propylène glycol (PG), méthanol (MeOH), ou chlorure de calcium (CaCl2)], furent évalués pour leur efficacité à tuer le virus de l'influenza aviaire (VIA) à −20 °C ou 21 °C. Une suspension de VIA fut appliquée à des disques d'acier inoxydable, séchés à l'air, et recouverts avec un désinfectant ou d'antigel pour une durée de 5 à 30 minutes. Le Virkon (2 %) et l'Accel (6,25 %) avec 30 % PG, 20 MeOH, ou 20 % CaCl2 ont inactivé 6 log10 de VIA en-dedans de 5 min à −20 °C et 21 °C. À ces températures le PG et le MeOH seuls n'ont pas réussi à tuer le VIA, mais la solution de CaCl2 à 20 % seule a inactivé 5 log10 de VIA en-dedans de 10 min. Les résultats suggèrent que le CaCl2 est potentiellement utile pour augmenter l'efficacité de la désinfection des exploitations avicoles suite à des poussées de cas d'infection par le VIA en saison chaude ou froide.(Traduit par Docteur Serge Messier).

Bacillus anthracis spore decontamination in food grease.

Bacillus anthracis Sterne strain spores were analyzed for their resistance against five disinfectants: commercial sodium hypochlorite, Spor-Klenz Ready-to-Use Cold Sterilant, accelerated hydrogen peroxide (AHP), Virkon, and surface decontamination foam (SDF). The aim of this study was to find an effective disinfectant that would reduce the viability of B. anthracis Sterne spores at ≥6 log in the presence of variables such as animal grease and fat, stainless steel, and temperature (room temperature and 4 °C). SDF and 10% sodium hypochlorite consistently reduced the growth of viable B. anthracis Sterne spores after 5 min in the presence of stainless steel at room temperature. It took at least 10 min of contact time for AHP to consistently reduce spore growth by ≥6 log, while it took at least 20 min for 5% bleach and Spor-Klenz to consistently inactivate ≥6 log spores in the presence of stainless steel at room temperature. AHP was the only disinfectant that reduced the viability of B. anthracis Sterne spores at ≥6 log in the presence of stainless steel and animal grease, both at room temperature and 4 °C after 24 h of contact time.

An investigation into human pandemic influenza virus (H1N1) 2009 on an Alberta swine farm.

On May 2, 2009 the Canadian Food Inspection Agency notified the World Organization for Animal Health that an emerging novel influenza A virus (pandemic H1N1 2009) had been confirmed on a swine farm in Alberta. Over a 4-week period pigs in this farrow-to-finish operation were clinically affected by respiratory disease consistent with an influenza A virus infection and the presence of active viral infection was confirmed in all production areas by real-time polymerase chain reaction (RT-PCR). Despite clinical recovery of animals, there was reluctance by purchasers to receive animals from this operation due to concerns about the effect on both domestic and international markets. The owner decided to depopulate the entire herd due to impending welfare issues associated with overcrowding and economic concerns resulting from the inability to market these animals. Carcasses were rendered or composted and did not enter the human food or animal feed chain. The source of virus in this herd was determined to be an infected human. Zoonotic transmission to 2 individuals responding to the outbreak was suspected and recommendations to prevent occupational exposure are discussed.