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Evaluation of a modified cleaning procedure in the prevention of carbapenem-resistant Acinetobacter baumannii clonal spread in a burn intensive care unit using a high-sensitivity luminometer

BACKGROUND Enhanced environmental cleaning practices are among the most accepted measures for controlling the spread of carbapenem-resistant Acinetobacter baumannii (CR-Ab). AIM To evaluate the impact of heightened cleaning on an ongoing CR-Ab outbreak in a burn intensive care unit (BICU) of an Italian teaching hospital, where chlorhexidine-60% isopropyl alcohol was applied as a complementary disinfectant on high-touch surfaces. METHODS Compliance with the microbial limit proposed for the BICU by AFNOR-NF-S90-351 (20 colony-forming units/100cm2) was assessed by plate count, and compared with the results obtained with intracellular adenosine triphosphate (ATP) detection. Genotyping was performed using pulsed-field gel electrophoresis. FINDINGS During the standard cleaning regimen, three out of 23 samples (13%) gave results over the AFNOR limit and five (21.7%) showed unacceptable ATP levels with 100 relative light units/100cm2 as the benchmark limit (sensibility 86.4%, specificity 92.2%). Following improvement of the cleaning procedure, only two samples out of 50 (4%) did not satisfy the microbiological criteria and seven (14%) exceeded the ATP limit. In a successive phase, eight of 30 samples collected showed unacceptable results (27%). CONCLUSIONS Adding chlorhexidine-60% isopropyl alcohol as complementary disinfectant proved to be effective for reducing environmental microbial contamination, ATP levels and CR-Ab infection/colonization in patients admitted to the BICU. Real-time monitoring by ATP assay was useful for managing the cleaning schedule and reducing hospital infections, although the calculated values must be interpreted as cleanliness indicators rather than risk indicators.

Colonization by Legionella spp. of water networks in residential buildings of the Province of Pisa, Italy

BACKGROUND Despite the increase of community acquired cases of legionellosis in Italy over the last years, the Italian guidelines do not give indications for prevention and control of Legionella in the hot water networks (or centralized conditioning systems) of residential buildings. We performed a survey on eight medium sized apartment buildings in the Pisa district to assess the prevalence of Legionella spp. in the water network and the respondance to drinking water requisites at the point of use, according to the Italian norms. METHODS For each building two hot water and three cold water samples (located at water entrance from the aqueduct network into the building pipework, at the exit from pressure autoclave, and at a remote tap) were collected. RESULTS Legionella was detected in 20% of residential buildings, mostly in those with a central hot water production system. CONCLUSIONS The study highlights a condition of potential risk for susceptible population subgroups and supports the need for measures of risk assessment and control.

Application of Hydrogen Peroxide as an Innovative Method of Treatment for Legionella Control in a Hospital Water Network

Objectives: To evaluate the effectiveness of hydrogen peroxide (HP) use as a disinfectant in the hospital water network for the control of Legionella spp. colonization. Methods: Following the detection of high levels of Legionella contamination in a 136-bed general hospital water network, an HP treatment of the hot water supply (25 mg/L) was adopted. During a period of 34 months, the effectiveness of HP on Legionella colonization was assessed. Legionella was isolated in accordance with ISO-11731 and identification was carried out by sequencing of the mip gene. Results: Before HP treatment, L. pneumophila sg 2–15 was isolated in all sites with a mean count of 9950 ± 8279 cfu/L. After one-month of HP treatment, we observed the disappearance of L. pneumophila 2–15, however other Legionella species previously not seen were found; Legionella pneumophila 1 was isolated in one out of four sampling sites (2000 cfu/L) and other non-pneumophila species were present in all sites (mean load 3000 ± 2887 cfu/L). Starting from September 2013, HP treatment was modified by adding food-grade polyphosphates, and in the following months, we observed a progressive reduction of the mean load of all species (p < 0.05), resulting in substantial disappearance of Legionella colonization. Conclusion: Hydrogen peroxide demonstrated good efficacy in controlling Legionella. Although in the initial phases of treatment it appeared unable to eliminate all Legionella species, by maintaining HP levels at 25 mg/L and adding food-grade polyphosphates, a progressive and complete control of colonization was obtained.

Evaluation of the Cleaning Procedure Efficacy in Prevention of Nosocomial Infections in Healthcare Facilities Using Cultural Method Associated with High Sensitivity Luminometer for ATP Detection

In healthcare facilities, environmental surfaces may be a reservoir of infectious agents even though cleaning and disinfection practices play a role in the control of healthcare-associated infections. In this study, the effectiveness of cleaning/disinfection procedures has been evaluated in two hospital areas, which have different risk category classifications. According to the contract with the cleaning service, after the daily ambulatory activities, the housekeeping staff apply an alcohol-based detergent followed by a chlorine-based disinfectant (2% Antisapril, Angelini; 540 mg/L active chlorine), properly diluted and sprayed. The contract provides for the use of disposable microfiber wipes which must be replaced with new ones in each health out-patient department. Surface contamination was analyzed using cultural methods and ATP detection, performed with a high-sensitivity luminometer. The values 100 CFU/cm2 and 40 RLU/cm2 were considered as the threshold values for medium-risk category areas, while 250 CFU/cm2 and 50 RLU/cm2 were defined for the low-risk category ones. Air quality was evaluated using active and passive sampling microbiological methods and particle count (0.3 μm–10 μm) detection. The cleaning/disinfection procedure reduced the medium bacterial counts from 32 ± 56 CFU/cm2 to 2 ± 3 CFU/cm2 in the low-risk area and from 25 ± 40 CFU/cm2 to 7 ± 11 CFU/cm2 in the medium-risk one. Sample numbers exceeding the threshold values decreased from 3% and 13% to 1% and 5%, respectively. RLU values also showed a reduction in the samples above the thresholds from 76% to 13% in the low-risk area. From the air samples collected using the active method, we observed a reduction of 60% in wound care and 53% in an ambulatory care visit. From the air samples collected using the passive method, we highlighted a 71.4% and 50% reduction in microbial contamination in the medium-risk area and in the low-risk one, respectively. The 10 μm size particle counts decreased by 52.7% in wound care and by 63% in the ambulatory care visit. Correct surface sanitation proved crucial for the reduction of microbial contamination in healthcare settings, and plays an important role in ensuring air quality in hospital settings.

Assessing natural mineral water microbiology quality in the absence of cultivable pathogen bacteria

Italian Directives recommend the good quality of natural mineral waters but literature data assert a potential risk from microorganisms colonizing wellsprings and mineral water bottling plants. We evaluated the presence of microorganisms in spring waters (SW) and bottled mineral waters (BMW) samples. Routine microbiological indicators, additional microorganisms like Legionella spp., Nontuberculous mycobacteria (NTM) and amoebae (FLA) were assessed in 24 SW and 10 BMW samples performing cultural and molecular methods. In 33 out of 34 samples, no cultivable bacteria ≥10 CFU/L was found. Cultivable FLA were detected in 50% of water samples. qPCR showed the presence of Legionella qPCR units in 24% of samples (from 1.1 × 102 to 5.8 × 102 qPCR units/L) and NTM qPCR units in 18% of samples (from 1 × 102 to 1 × 105 qPCR units/L). Vermamoeba vermiformis and Acanthamoeba polyphaga were recovered respectively in 70% of BMW samples (counts from 1.3 × 103 to 1.2 × 105 qPCR units/L) and 42% of SW samples (from 1.1 × 103 to 1.3 × 104 qPCR units/L). Vahlkampfia spp. was detected in 42% of SW and 70% of BMW samples (from 1.2 × 103 to 1.2 × 105 qPCR units/L). Considering the presence of FLA, we underline the importance of a wider microbiological risk assessment in natural mineral waters despite the absence of cultivable bacteria.

Improving Cleaning and Disinfection of High-Touch Surfaces in Intensive Care during Carbapenem-Resistant Acinetobacter baumannii Endemo-Epidemic Situations

Aims: High-touch surfaces cleaning and disinfection require the adoption of effective and proper executed protocols, especially during carbapenem-resistant Acinetobacter baumannii (CRAB) endemo-epidemic situations. We evaluated the effectiveness and residual disinfectant activity of disposable pre-impregnated wipes (Modified Operative Protocol, MOP) in reducing environmental bioburden versus a two-step Standard Operative Protocol (SOP) in a 12-bed Intensive Care Unit. Methods: Five high-touch surfaces were cleaned and disinfected either according to the SOP (alcohol-based cleaning and chlorine-based disinfection) or using quaternary ammonium compounds-based disposable wipes (MOP). Sampling was performed before each procedure and at 0.5, 2.5, 4.5 and 6.5 h after (560 sites). Total viable count (TVC) was evaluated according to Italian hygiene standard (<50 CFU/24 cm2). Clinical and environmental CRAB strains isolated were genotyped. Results: On non-electromedical surfaces the difference between TVC before procedure and at each of the following times was significant only for the MOP (p < 0.05, Wilcoxon test). Using the MOP, only 7.4% (10/135) of sites showed TVC >50 CFU/24 cm2 (hygiene failures) versus 18.9% (25/132) after SOP (p < 0.05, Fisher’s Exact test). On infusion pumps a higher number of hygiene failures was observed after the SOP (7/44, 15.9%) compared with the MOP (4/45, 8.9%). Genotyping highlighted a common source of infection. Conclusion: On high-touch surfaces, the use of disposable wipes by in-house auxiliary nurses may represent a more effective alternative to standard cleaning and disinfection procedure performed by outsourced cleaning services, showing effectiveness in reducing microbial contamination and residual disinfection activity up to 6.5 h.

Evaluation and control of microbial and chemical contamination in dialysis water plants of Italian nephrology wards

BACKGROUND Patients receiving haemodialysis are exposed to a large volume of dialysis fluid. The Italian Society of Nephrology (ISN) has published guidelines and microbial quality standards on dialysis water (DW) and solutions to ensure patient safety. AIM To identify microbial and chemical hazards, and evaluate the quality of disinfection treatment in DW plants. METHODS In 2015 and 2016, water networks and DW plants (closed loop and online monitors) of nine dialysis wards of Italian hospitals, hosting 162 dialysis beds overall, were sampled on a monthly basis to determine the parameters provided by ISN guidelines. Chlorinated drinking water was desalinated by reverse osmosis and distributed to the closed loop which feeds all online monitors. Disinfection with peracetic acid was performed in all DW plants on a monthly basis. FINDINGS Over the 24-month study period, seven out of nine DW plants (78%) recorded negative results for all investigated parameters. Closed loop contamination with Burkholderia cepacia was detected in a DW plant from January 2015 to March 2015. Pseudomonas aeruginosa was isolated from March 2016 to May 2016 in the closed loop of another DW plant. These microbial contaminations were eradicated by shock disinfection with sodium hypochlorite and peracetic acid, followed by water flushing. CONCLUSION These results highlight the importance of chemical and physical methods of DW disinfection. The maintenance of control measures in water plants hosted in dialysis wards ensures a microbial risk reduction for all dialysis patients.

Presence of Legionella spp. in Hot Water Networks of Different Italian Residential Buildings: A Three-Year Survey

Although the European reports highlight an increase in community-acquired Legionnaires’ disease cases, the risk of Legionella spp. in private houses is underestimated. In Pisa (Italy) we performed a three-year survey on Legionella presence in 121 buildings with an independent hot water production (IB); 64 buildings with a central hot water production (CB); and 35 buildings with a solar thermal system for hot water production (TB). From all the 220 buildings Legionella spp. was researched in two hot water samples collected either at the recirculation point or on the first floor and on the last floor, while the potable water quality was analysed in three cold water samples collected at the inlet from the aqueduct network, at the exit from the autoclave, and at the most remote tap. Legionella pneumophila sg1, Legionella pneumophila sg2–16, and non-pneumophila Legionella species were detected in 26% of the hot water networks, mostly in CB and TB. In these buildings we detected correlations between the presence of Legionella and the total chlorine concentration decrease and/or the increase of the temperature. Cold water resulted free from microbiological hazards, with the exception of Serratia liquefaciens and Enterobacter cloacae isolated at the exit from two different autoclaves. We observed an increase in total microbial counts at 22 °C and 37 °C between the samples collected at the most remote taps compared to the ones collected at the inlet from the aqueduct. The study highlights a condition of potential risk for susceptible categories of population and supports the need for measures of risk assessment and control.