Modeling microbial survival in buildup biofilm for complex medical devices.
BACKGROUND
Flexible endoscopes undergo repeated rounds of patient-use and reprocessing. Some evidence indicates that there is an accumulation or build-up of organic material that occurs over time in endoscope channels. This "buildup biofilm" (BBF) develops as a result of cyclical exposure to wet and dry phases during usage and reprocessing. This study investigated whether the BBF matrix represents a greater challenge to disinfectant efficacy and microbial eradication than traditional biofilm (TBF), which forms when a surface is constantly bathed in fluid.


METHODS
Using the MBEC (Minimum Biofilm Eradication Concentration) system, a unique modelling approach was developed to evaluate microbial survival in BBF formed by repetitive cycles of drying, disinfectant exposure and re-exposure to the test organism. This model mimics the cumulative effect of the reprocessing protocol on flexible endoscopes. Glutaraldehyde (GLUT) and accelerated hydrogen peroxide (AHP) were evaluated to assess the killing of microbes in TBF and BBF.


RESULTS
The data showed that the combination of an organic matrix and aldehyde disinfection quickly produced a protective BBF that facilitated high levels of organism survival. In cross-linked BBF formed under high nutrient conditions the maximum colony forming units (CFU) reached ~6 Log(10)CFU/peg. However, if an oxidizing agent was used for disinfection and if organic levels were kept low, organism survival did not occur. A key finding was that once established, the microbial load of BBF formed by GLUT exposure had a faster rate of accumulation than in TBF. The rate of biofilm survival post high-level disinfection (HLD) determined by the maximum Log(10)CFU/initial Log(10)CFU for E. faecalis and P. aeruginosa in BBF was 10 and 8.6 respectively; significantly different compared to a survival rate in TBF of ~2 for each organism. Data from indirect outgrowth testing demonstrated for the first time that there is organism survival in the matrix. Both TBF and BBF had surviving organisms when GLUT was used. For AHP survival was seen less frequently in BBF than in TBF.


CONCLUSIONS
This BBF model demonstrated for the first time that survival of a wide range of microorganisms does occur in BBF, with significantly more rapid outgrowth compared to TBF. This is most pronounced when GLUT is used compared to AHP. The data supports the need for meticulous cleaning of reprocessed endoscopes since the presence of organic material and microorganisms prevents effective disinfection when GLUT and AHP are used. However, cross-linking agents like GLUT are not as effective when there is BBF. The data from the MBEC model of BBF suggest that for flexible endoscopes that are repeatedly used and reprocessed, the assurance of effective high-level disinfection may decrease if BBF develops within the channels.
Department of Medical Microbiology, University of Manitoba, Winnipeg, MB, Canada. malfa@sbgh.mb.ca
Michelle J
MJ
Alfa
Rosemarie
R
Howie
D000704:Analysis of Variance; D018441:Biofilms / Q000145:classification / Q000254:growth & development*; D002176:Candida albicans / Q000254:growth & development; D015169:Colony Count, Microbial; D003470:Culture Media; D004203:Disinfection / Q000379:methods*; D019723:Endoscopes / Q000382:microbiology*; D013293:Enterococcus faecalis / Q000254:growth & development; D004866:Equipment Contamination; D017743:Equipment Reuse; D005976:Glutaral; D006861:Hydrogen Peroxide; D050296:Microbial Viability; D008954:Models, Biological; D011550:Pseudomonas aeruginosa / Q000254:growth & development
D003470:Culture Media; D006861:Hydrogen Peroxide; D005976:Glutaral