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To evaluate the efficacy of detergent and friction on removal of traditional biofilm and cyclic-buildup biofilm (CBB) from polytetrafluoroethylene (PTFE) channels and to evaluate the efficacy of glutaraldehyde to kill residual bacteria after cleaning.
PTFE channels were exposed to artificial test soil containing 108 CFU/mL of Pseudomonas aeruginosa and Enterococcus faecalis, followed by full cleaning and high-level disinfection (HLD) for five repeated rounds to establish CBB. For traditional biofilm, the HLD step was omitted. Cleaning with enzymatic and alkaline detergents, bristle brush, and Pull Thru channel cleaner were compared to a water flush only. Carbohydrate, protein, viable count, adenosine triphosphate (ATP) levels were analyzed and atomic force microscopy (AFM) was performed.
In the absence of friction, cleaning of traditional biofilm and CBB was not effective compared to the positive control (Dunn-Bonferroni tests; P > .05) regardless of the detergent used. ATP, protein, and carbohydrate analyses were unable to detect traditional biofilm or CBB. The AFM analysis showed that fixation resulted in CBB being smoother and more compact than traditional biofilm.
Friction during the cleaning process was a critical parameter regardless of the detergent used for removal of either traditional biofilm or CBB. Glutaraldehyde effectively killed the remaining microorganisms regardless of the cleaning method used.
Biofilm has been implicated in bacterial persistence and survival after endoscope reprocessing. In this study, we assessed the impact of different methods of reprocessing on organic residues and viable bacteria after repeated rounds of biofilm formation when each was followed by full reprocessing.
ATS-2015, an artificial test soil containing 5–8 Log10 colony-forming units (CFU) of Enterococcus faecalis and Pseudomonas aeruginosa, was used to form biofilm in polytetrafluroethylene channels overnight on 5 successive days. Each successive day, full pump-assisted cleaning using bristle brushes or pull-through devices in combination with enzymatic or nonenzymatic detergents followed by fully automated endoscope reprocessor disinfection using peracetic acid was performed. Residuals were visualized by scanning electron microscopy (SEM). Destructive testing was used to assess expected cutoffs for adenosine triphosphate (ATP; <200 relative light units), protein (<2 µg/cm2), and viable bacteria count (0 CFU).
Protein residuals were above 2 µg/cm2, but ATP residuals were <200 relative light units for all methods tested. Only when enzymatic cleaner was used for cleaning were there no viable bacteria detected after disinfection irrespective of whether bristle brushes or pull-through devices were used. SEM revealed that some residual debris remained after all reprocessing methods, but more residuals were detected when a nonenzymatic detergent was used.
Surviving E. faecalis and P. aeruginosa were only detected when the non-enzymatic detergent was used, emphasizing the importance of the detergent used for endoscope channel reprocessing. Preventing biofilm formation is critical because not all current reprocessing methods can reliably eliminate viable bacteria within the biofilm matrix.
The primary objective of this study was to evaluate fluorescent readout results of Attest 1291 Biological Indicators (Bis) (3M Health Care, St. Paul, MN) and Attest 1296 BI test packs (containing Attest 1292 Bis) using full and fractional cycles compared with the growth data when prolonged incubation (7 days) was included. Gravity displacement and vacuum-assisted steam sterilization cycles were evaluated. A secondary objective of this study was to evaluate the new automated rapid fluorescent reader (Attest 290 Auto Reader).
The rapid readout Bis for gravity displacement and vacuum-assisted steam autoclave cycles at 132° C were processed using full (4 minutes) and four fractional cycles that provided 30% to 80% positive results for growth after 24 hours of incubation (48 hours of incubation for Attest 1292 Bis from the Attest 1296 test packs). Sixty of each type of BI were tested for each cycle (300 of each BI type in total).
For all full steam sterilization cycles, results of the rapid fluorescent readout and the 24-hour, 48-hour, and 7-day growth tests were negative for all Attest 1291 and 1292 Bis tested. For all fractional cycles, the 24- and 48-hour growth results for the Attest 1291 and 1292 Bis, respectively, were the same as the 7-day growth results. The fractional cycle data indicated that fluorescent rapid readout was a more sensitive indicator than growth. There were rare (0.9%) false-negative results for Bis under fractional cycle conditions and these all correlated with short fractional cycle exposure times.
The fluorescent rapid readout results of the 1291 Bis and 1296 BI test packs reliably predict both 24- and 48-hour and 7-day growth. These data support the value of rapid readout Bis for sterilizer monitoring for both the vacuum-assisted and the gravity displacement steam sterilization cycles. The new automated reader requires less manipulation of the BI and makes monitoring user friendly and less prone to user errors.
To obtain information about current reprocessing practices and to obtain samples from the biopsy channel to quantitate soil levels and bioburden in patient-ready flexible duodenoscopes used for endoscopic retrograde choliangiopancreatography (ERCP).
Participating centers were sent a questionnaire and a kit for on-site collection of samples from the biopsy channel of the duodenoscope.
Thirty-seven hospitals from across Canada participated. The only criterion was that they currently used and reprocessed flexible duodenoscopes for ERCP procedures.
The questionnaire obtained information on reprocessing practices. The kit included a detailed instruction booklet outlining sample collection and all of the tubes, sterile water, and brushes needed for it. Samples were collected on-site from all ERCP scopes in each center on Monday morning and shipped by overnight courier on ice to the research center. Each sample was assayed by routine microbiologic methods for total viable count and protein, blood, carbohydrate, and endotoxin levels.
Microbial overgrowth was present in 7% of 119 scope samples. Cleaning appeared to be reasonably well done in most of the centers, and 43% of the centers were in total compliance with basic national guidelines. The data from the scope samples indicated that there was significantly greater buildup of protein, carbohydrate, and endotoxin associated with ERCP scopes from centers using glutaraldehyde, compared with those using peracetic acid. Carbohydrate was the soil component detected most frequently and in the highest concentration in scope channels.
Although cleaning was generally well done, areas for improvement included ensuring the availability of written reprocessing protocols, immersion of scopes during manual cleaning, use of adequate fluid volume for rinsing, adequate drying of scopes prior to storage, and the separation of ERCP valves from scopes during storage.
To use a serum and salt challenge in narrow-lumen carriers to evaluate a 10% ethylene oxide plus 90% hydrochlorofluorocarbon (EO-HCFC) sterilant mixture in a retrofitted 12/88 sterilizer as an alternative to the banned chlorofluorocarbon-ethylene oxide (EO) sterilant mixture.
An EO-HCFC sterilizing gas mixture in a retrofitted 12/88 sterilizer was compared to 100% ethylene oxide (100% EO) sterilizing gas to determine its relative ability to kill seven different bacteria (Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Bacillus subtilis spores, Bacillus sUawthermophilus spores, Bacillus circulans spores, and Mycobacterium chelonei) in the presence or absence of a combined 10% serum and 0.65% salt challenge using both penicylinders (PC) and long narrowlumen (LU) carriers.
The EO-HCFC sterilant mixture (96% sterile carriers) was equivalent to the 100% EO sterilant (98% sterile carriers) for killing vegetative organisms, as well as spore suspensions, on the 27 PC and 27 LU carriers in the absence of serum and salt. In the presence of serum and salt, the EO-HCFC sterilant mixture was markedly better than the 100% EO sterilant at reducing the bacterial load on the 63 PC carriers (95% vs 62% sterile PC carriers, respectively), whereas both sterilizers were equivalent for the 63 LU carriers (49% vs 40% sterile LU carriers, respectively). Of the seven test organisms, E faecalis, B subtilis, B stearothermophilus, and B circulans were the most difficult to kill for both PC and LU carriers when serum and salt were present.
The data presented in this report indicate that the EO-HCFC sterilant mixture is an effective alternative for gas sterilization. Indeed, the efficiency of bacterial killing for the EO-HCFC sterilant mixture was similar to that achieved by the 12/88 EO-CFC sterilant mixture.
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