Burkholderia cepacia-contaminated dentures possibly cause pneumonia.

We report a case in which a patient was suspected of developing pneumonia due to wearing dentures that were immersed in a storage solution contaminated with 3.0 × 108 colony-forming units (cfu)/mL of Burkholderia cepacia. It is highly possible that the contaminated denture solution entered the trachea and caused the pneumonia, possibly due to the prolonged supine positioning of the patient. We demonstrated that B. cepacia isolated from the sputum and B. cepacia isolated from the denture storage solution had the same DNA fingerprint, and that the patient recovered from pneumonia after stopping the use of dentures. These findings suggest the storage solution as the main source of infection.

Peripheral Parenteral Nutrition Solutions and Bed Bath Towels as Risk Factors for Nosocomial Peripheral Venous Catheter-related Bloodstream Infection by Bacillus cereus.

In Japan, China, and Singapore, several studies have reported increased incidences of peripheral venous catheter-related bloodstream infection by Bacillus cereus during the summer. Therefore, we hypothesized that bed bathing with a B. cereus-contaminated "clean" towels increases B. cereus contact with the catheter and increases the odds of contaminating the peripheral parenteral nutrition (PPN). We found that 1) professionally laundered "clean" towels used in hospitals have B. cereus (3.3×104 colony forming units (CFUs) / 25cm2), 2) B. cereus is transferable onto the forearms of volunteers by wiping with the towels (n=9), and 3) B. cereus remain detectable (80∼660 CFUs /50cm2) on the forearms of volunteers even with subsequent efforts of disinfection using alcohol wipes. We further confirmed that B. cereus grow robustly (102 CFUs /mL to more than 106 CFUs /mL) within 24hours at 30°C in PPN. Altogether we find that bed bathing with a towel contaminated with B. cereus leads to spore attachments to the skin, and that B. cereus can proliferate at an accelerated rate at 30°C compared to 20°C in PPN. We therefore highly recommend ensuring the use of sterile bed bath towels prior to PPN administration with catheter in patients requiring bed bathing.

Possibility of a case of pneumonia caused by green tea contaminated with Pseudomonas aeruginosa and countermeasures against it.

We report a patient who developed pneumonia after prolonged use of spray bottle containing green tea for hydration purposes. The cause was suspected to be a contamination of green tea because the patient's symptoms persisted and did not improve until stopping the use of the spray bottle and we also found the green tea in the spray bottle to harbor a high number of Pseudomonas aeruginosa (1.2 × 107 colony forming units (cfu)/mL). It is not uncommon to use green tea for hydration or gargling purposes in some patient care settings considering the antibacterial effects of catechins contained in green tea. Our findings suggest the importance of keeping vigilance on consuming green tea in spray bottles in hospital settings since it may readily be contaminated by pathogens such as P. aeruginosa.

Microbial contamination of hands with or without the use of bidet toilets (electric toilet seats with water spray) after defecation.

Bidet toilets (electric toilet seats with water spray) are increasing in popularity worldwide. However, the extent of reduction of microbial contamination of the hands with the use of bidet toilets after defecation is unclear. Microbe contamination of the hands with and without the use of bidet toilets after defecation was examined in 32 nursing students. Double gloves were worn on the dominant hand and four layers of toilet paper were used to wipe the buttocks after defecation, and microbe contamination of the second glove (outer glove) of the double gloves was examined. The volunteers were free to select the flow volume, wash time of the bidet, and the type of bidet. Without the use of a bidet toilet, the average value ± standard deviation of the number of microbes attached to the gloves was 39,499.3 ± 77,768.3 colony forming units (cfu)/glove; however, it was 4,146.9 ± 11,427.7 cfu/glove when the bidet toilet was used. The number of microbes adhering to gloves was significantly reduced when a bidet toilet was used (p < 0.00001, Wilcoxon signed-rank test).

Rapid Detection of Microbial Contamination in Intravenous Fluids by ATP-Based Monitoring System.

In this study, we investigated four clinical cases of microbial contamination of in-use intravenous infusion fluid, detected by measuring "Adenosine triphosphate (ATP) + adenosine monophosphate (AMP)" ("ATP+AMP") levels. High "ATP+AMP" values correlate with microbial contamination, and by utilizing these values as indicator for microbial contamination possibility, we were able to rapidly detect the contamination and recommend replacement of catheters and administration sets. In three out of four cases, changing the infusion fluid led to improvement in the condition of the patients. "ATP+AMP" levels can be used to confirm microbial contamination of in-use intravenous infusion fluids, as it is fast (several minites) and convenient to measure them.

The Use of Electric Toilet Seats with Water Spray Is Efficacious in Maintaining Hand Hygiene in Experimental Model.

A simulation experiment was conducted to examine hand contamination from wiping the buttocks after the use and non-use of an electric toilet seat with water spray. A model of the buttocks was smeared with artificial diarrheal feces containing Serratia marcescens, and wiped by the participants wearing disposable gloves with 4 sheets of toilet paper after the use and non-use of the water spray of an electric toilet seat. Subsequently, the presence of S. marcescens on the surface of the gloves was quantified. After using the water spray, the mean count±standard deviation of S. marcescens was 0.067±0.249 colony-forming units (cfu)/glove, and it was 4,275±6,069 cfu/glove when water spray was not used. The cfu of S. marcescens was significantly lower when the water spray was used (p<0.00001) prior to wiping the artificial diarrheal feces. This result supports the effectiveness of water spray to prevent defecation-related hand contamination.

The Risk of Microbial Contamination in Multiple-Dose Preservative-Free Ophthalmic Preparations.

Multiple-dose ophthalmic preparations that do not contain preservatives carry high risks of microbial contamination. However, there are various types of hospital preparations, with different physicochemical properties. In the present study, we evaluated the association between physicochemical properties and microbial contamination in ophthalmic preparations. The investigated hospital preparations included ophthalmic preparations of physiological saline, 0.2% fluconazole, 0.5% vancomycin hydrochloride, and 2% cyclosporine. We investigated the microbial dynamics of each ophthalmic preparation and microbial contamination in ophthalmic preparations used by patients. Remarkable growth of Pseudomonas aeruginosa, Burkholderia cepacia, and Serratia marcescens was observed in ophthalmic preparations of physiological saline and 0.2% fluconazole. All tested microorganisms displayed decreased counts after inoculation in 0.5% vancomycin hydrochloride. In 2% cyclosporine, all investigated microorganisms were below the limit of detection after inoculation for 6 h. The microbial contamination rates of ophthalmic preparations used by patients were 16.7% (3/18 samples) for 0.5% vancomycin hydrochloride and 0% (0/30 samples) for 2% cyclosporine. All detected contaminants in 0.5% vancomycin hydrochloride were Candida spp., one of which was present at a level of 1×104 colony-forming units/mL. The storage method for in-use ophthalmic preparations should be considered on the basis of their physicochemical properties.

Contamination of environmental surfaces by methicillin-resistant Staphylococcus aureus (MRSA) in rooms of inpatients with MRSA-positive body sites

ABSTRACT Methicillin-resistant Staphylococcus aureus (MRSA) can contaminate environmental surfaces that are frequently touched by the hands of patients with MRSA colonization/infection. There have been many studies in which the presence or absence of MRSA contamination was determined but no studies in which MRSA contamination levels were also evaluated in detail. We evaluated MRSA contamination of environmental surfaces (overbed tables, bed side rails, and curtains) in the rooms of inpatients from whom MRSA was isolated via clinical specimens. We examined the curtains within 7-14 days after they had been newly hung. The environmental surfaces were wiped using gauze (molded gauze for wiping of surface bacteria; 100% cotton, 4 cm × 8 cm) moistened with sterile physiological saline. The MRSA contamination rate and mean counts (range) were 25.0% (6/24 samples) and 30.6 (0-255) colony-forming units (cfu)/100 cm2, respectively, for the overbed tables and 31.6% (6/19 samples) and 159.5 (0-1620) cfu/100 cm2, respectively, for the bed side rails. No MRSA was detected in 24 curtain samples. The rate of MRSA contamination of environmental surfaces was high for the overbed tables and bed side rails but low for the curtains. Therefore, at least until the 14th day of use, frequent disinfection of curtains may be not necessary.

Microbial Contamination of Ice Machines Is Mediated by Activated Charcoal Filtration Systems in a City Hospital.

Although microbial contamination of ice machines has been reported, no previous study has addressed microbial contamination of ice produced by machines equipped with activated charcoal (AC) filters in hospitals. The aim of this study was to provide clinical data for evaluating AC filters to prevent microbial contamination of ice. We compared microbial contamination in ice samples produced by machines with (n = 20) and without an AC filter (n = 40) in Shunan City Shinnanyo Municipal Hospital. All samples from the ice machine equipped with an AC filter contained 10-116 CFUs/g of glucose nonfermenting gram-negative bacteria such as Pseudomonas aeruginosa and Chryseobacterium meningosepticum. No microorganisms were detected in samples from ice machines without AC filters. After the AC filter was removed from the ice machine that tested positive for Gram-negative bacteria, the ice was resampled (n = 20). Analysis found no contaminants. Ice machines equipped with AC filters pose a serious risk factor for ice contamination. New filter-use guidelines and regulations on bacterial detection limits to prevent contamination of ice in healthcare facilities are necessary.

Contamination of environmental surfaces by methicillin-resistant Staphylococcus aureus (MRSA) in rooms of inpatients with MRSA-positive body sites.

Methicillin-resistant Staphylococcus aureus (MRSA) can contaminate environmental surfaces that are frequently touched by the hands of patients with MRSA colonization/infection. There have been many studies in which the presence or absence of MRSA contamination was determined but no studies in which MRSA contamination levels were also evaluated in detail. We evaluated MRSA contamination of environmental surfaces (overbed tables, bed side rails, and curtains) in the rooms of inpatients from whom MRSA was isolated via clinical specimens. We examined the curtains within 7-14 days after they had been newly hung. The environmental surfaces were wiped using gauze (molded gauze for wiping of surface bacteria; 100% cotton, 4cm×8cm) moistened with sterile physiological saline. The MRSA contamination rate and mean counts (range) were 25.0% (6/24 samples) and 30.6 (0-255)colony-forming units (cfu)/100cm(2), respectively, for the overbed tables and 31.6% (6/19 samples) and 159.5 (0-1620)cfu/100cm(2), respectively, for the bed side rails. No MRSA was detected in 24 curtain samples. The rate of MRSA contamination of environmental surfaces was high for the overbed tables and bed side rails but low for the curtains. Therefore, at least until the 14th day of use, frequent disinfection of curtains may be not necessary.

Cleanliness of Linen and Clothing Items Professionally Laundered or Dry-Cleaned.

We asked 14 professional cleaners (laundry services) to clean various unused (new) linen and clothing items with a microbial contamination level of <1 cfu/cm(2) and then evaluated the bacterial/fungal contamination of the laundered or dry-cleaned items. After laundering, 6 (21.4%) of the 28 samples from 4 of the 14 cleaners (28.6%) were contaminated (1-1,200 cfu/cm(2)). After dry-cleaning, 2 (7.1%) of the 28 samples from 2 (14.3%) of the 14 cleaners were contaminated (7-10 cfu/cm(2)). The main contaminant was Bacillus cereus. No sample of the laundered or dry-cleaned items showed Staphylococcus aureus, Escherichia coli, or Pseudomonas aeruginosa contamination. All 14 cleaners investigated in this study used batch-type washing machines. Therefore, batch-type washing machines can cause contamination of linen and clothing items with B. cereus.

Efficacy of disinfectants against naturally occurring and artificially cultivated bacteria.

Naturally occurring bacteria, is exist in nature, and is never cultivated on conventional culture medium. We evaluated the efficacy of disinfectants against naturally occurring bacteria in in-use cotton balls soaked in 0.02% benzalkonium chloride solution which had been used to disinfect the genital area by patients undergoing self-catheterization at home and the same bacteria subcultured on nutrient broth (artificially cultivated bacteria). The colony forming units (CFU) of naturally occurring bacteria such as Serratia marcescens, Alcaligenes xylosoxidans, and Burkholderia cepacia were not decreased after 48 h exposure to 0.025-0.1% benzalkonium chloride solution, but the same strains subcultured on nutrient broth were killed within only 10 min exposure to 0.025-0.1% benzalkonium chloride solution. In addition, the CFU of these three kinds of naturally occurring bacteria were not decreased after 48 h exposure to 0.02% chlorhexidine gluconate solution, but the same strains subcultured on nutrient broth were killed within 2 h exposure to chlorhexidine gluconate solution. The result showed that disinfectant efficacy differed markedly against naturally occurring and artificially cultivated bacteria. Therefore, it is preferable to use the naturally occurring bacteria not only artificially cultivated bacteria when examining disinfectant efficacy.

Residual amount of vancomycin hydrochloride in vials after use.

We macroscopically observed vials of vancomycin hydrochloride (VCM) for injection (0.5 g/vial) dissolved in various solvents, and determined the presence or absence of residual VCM crystals. In addition, the residual VCM in vials after use was measured using a bioassay. In vials evaluated after use, the percentage of vials in which VCM crystals were macroscopically confirmed, the mean residual amount of VCM in the vials (residual %), and the percentage of vials with ≥50 mg (10 %) of residual VCM were 28.1 %, 15.0 ± 7.5 mg (3.0 %), and 0 %, respectively, after the dissolution of a single VCM vial in 10 ml of distilled water for injection (n = 57); 63.8 %, 30.2 ± 19.1 mg (6.0 %), and 16.1 %, respectively, after the dissolution of a single VCM vial in 100 ml of physiological saline (n = 224); and 72.2 %, 38.5 ± 28.0 mg (7.7 %), and 33.3 %, respectively, after the dissolution of two VCM vials in 100 ml of physiological saline (n = 18). The mean residual VCM amount was greater when using physiological saline than when using distilled water for injection as a solvent. These results show the need to follow the dissolution method described in the package insert, which calls for the addition of 10 ml of distilled water for injection to each 0.5 g VCM vial.

Disinfection methods for spores of Bacillus atrophaeus, B. anthracis, Clostridium tetani, C. botulinum and C. difficile.

To evaluate disinfection methods for environments contaminated with bioterrorism-associated microorganism (Bacillus anthracis), we performed the following experiments. First, the sporicidal effects of sodium hypochlorite on spores of five bacterial species were evaluated. Bacillus atrophaeus was the most resistant to hypochlorite, followed in order by B. anthracis, Clostridium botulinum and Clostridium tetani, and Clostridium difficile. Subsequently, using B. atrophaeus spores that were the most resistant to hypochlorite, the sporicidal effects of hypochlorite at lower pH by adding vinegar were evaluated. Hypochlorite containing vinegar had far more marked sporicidal effects than hypochlorite alone. Cleaning with 0.5% (5000 ppm) hypochlorite containing vinegar inactivated B. atrophaeus spores attached to vinyl chloride and plywood plates within 15 s, while that not containing vinegar did not inactivate spores attached to cement or plywood plates even after 1 h. Therefore, the surfaces of cement or plywood plates were covered with gauze soaked in 0.5% hypochlorite containing vinegar, and the sporicidal effects were evaluated. B. atrophaeus spores attached to plywood plates were not inactivated even after 6 h, but those attached to cement plates were inactivated within 5 min. On the other hand, covering the surfaces of plywood plates with gauze soaked in 0.3% peracetic acid and gauze soaked in 2% glutaral inactivated B. atrophaeus spores within 5 min and 6 h, respectively. These results suggest that hypochlorite containing vinegar is effective for disinfecting vinyl chloride, tile, and cement plates contaminated with B. anthracis, and peracetic acid is effective for disinfecting plywood plates contaminated with such microorganism.

Microbial contamination of disinfectants used for intermittent self-catheterization.

We investigated the microbial contamination of 0.02% benzalkonium chloride solution used in catheter kits for intermittent self-catheterization. Of 20 samples examined, 12 (60.0%) were contaminated with 8.8 x 10(2)-3.1 x 10(6) colony-forming units (cfu)/mL. The contaminants were Pseudomonas fluorescens, Burkholderia cepacia, and Aeromonas spp. These results showed that 0.02% benzalkonium chloride solution used for the lubrication/disinfection of catheters for self-catheterization is susceptible to contamination. Therefore, the lubricant/disinfectant for catheters for self-catheterization was changed from 0.02% benzalkonium chloride solution to 84-87% glycerin containing 0.02% benzalkonium chloride, and microbial contamination of the latter in catheter kits for self-catheterization was reinvestigated. Of 42 samples, 5 (11.9%) were contaminated with 20-2.0 x 10(4) cfu/mL. However, the rate of contamination of 84-87% glycerin containing 0.02% benzalkonium chloride was significantly lower than that of 0.02% benzalkonium chloride solution (P<0.0001). The contaminant of 84-87% glycerin containing 0.02% benzalkonium chloride was Bacillus spp. in all contaminated samples. In this survey, neither contaminants of 0.02% benzalkonium chloride solution nor the contaminant of 84-87% glycerin containing 0.02% benzalkonium chloride were the causative microbial species of urinary tract infection.

Microbial contamination of suction tubes attached to suction instruments and preventive methods.

We investigated the microbial contamination of suction tubes attached to wall-type suction instruments. Microbial contamination of suction tubes used for endoscopy or sputum suction in hospital wards was examined before and after their disinfection. In addition, disinfection and washing methods for suction tubes were evaluated. Suction tubes (n=33) before disinfection were contaminated with 10(2)-10(8) colony-forming units (cfu)/tube. The main contaminants were Pseudomonas aeruginosa, Acinetobacter baumannii, and Stenotrophomonas maltophilia. The suction tubes were disinfected with sodium hypochlorite (n=11) or hot water (n=11), or by an automatic tube cleaner (n=11). After 2-h immersion in 0.1% (1,000 ppm) sodium hypochlorite, 10(3)-10(7) cfu/tube of bacteria were detected in all 11 tubes examined. After washing in hot running water (65 degrees C), 10(3)-10(7) cfu/tube were detected in 3 of the 11 examined tubes. The bacteria detected in the suction tubes after disinfection with sodium hypochlorite or hot water were P. aeruginosa, A. baumannii, and S. maltophilia. On the other hand, after washing with warm water (40 degrees C) using the automatic tube cleaner, contamination was found to be <20 cfu/tube (lower detection limit, 20 cfu/tube) in all 11 tubes examined. These results suggest the usefulness of washing with automatic tube cleaners.

In vitro antimicrobial effects of aztreonam, colistin, and the 3-drug combination of aztreonam, ceftazidime and amikacin on metallo-beta-lactamase-producing Pseudomonas aeruginosa.

BACKGROUND: There are limited choice of antimicrobial agents to treat infection with metallo-beta-lactamase-producing Pseudomonas aeruginosa. We evaluate the antimicrobial effects of aztreonam alone, colistin alone and the 3-drug combination of aztreonam, ceftazidime and amikacin on 23 strains of metallo-beta-lactamase-producing P. aeruginosa by time-killing tests. METHODS: Strains used were from different hospitals in Japan and had different pulse-field gel electrophoresis patterns by restriction with SpeI. The minimum inhibitory concentrations of 11 antimicrobial agents (piperacillin, piperacillin/tazobactam, imipenem, meropenem, aztreonam, ceftazidime, amikacin, tobramycin, arbekacin, ciprofloxacin and colistin) were determined using the agar dilution test. The effects of aztreonam, colistin and the combination of aztreonam, ceftazidime and amikacin were determined by time-killing studies. RESULTS: Bacteriostatic effects after 6 hours of drug exposure were observed in 12 strains (52.2%) of 23 strains of metallo-beta-lactamase-producing P. aeruginosa with 48 mg/l aztreonam, in 19 strains (82.6%) with the 3-drug combination of 16 mg/l aztreonam, 16 mg/l ceftazidime, and 4 mg/l amikacin, and in 23 strains (100%) with 2 mg/l colistin. Bactericidal effects after 6 h drug exposure were observed in 1 strain (4.3%) with 48 mg/l aztreonam, in 8 strains (30.4%) with the 3-drug combination and in all 23 strains (100%) with 2 mg/l colistin. CONCLUSION: Evaluation of in vitro antimicrobial effects on metallo-beta-lactamase-producing P. aeruginosa revealed relatively good effects of the 3-drug combination of aztreonam, ceftazidime and amikacin and marked effects of colistin.

Antimicrobial effects of drugs against multidrug-resistant Pseudomonas aeruginosa.

We evaluated the effects of antimicrobial drugs on four strains of Pseudomonas aeruginosa that are resistant to eight widely used antipseudomonal drugs (piperacillin, piperacillin-tazobactam, imipenem, meropenem, ceftazidime, aztreonam, amikacin, ciprofloxacin) and colistin. In the killing test, colistin (2 microg/ml) was the most effective, followed by aztreonam (48 microg/ml), piperacillin-tazobactam (192-4 microg/ml), piperacillin (192 microg/ml), and a three drug combination of azetreonam (16 microg/ml), ceftazidime (16 microg/ml), and amikacin (4 microg/ml). Six hours after drug addition, colistin (2 microg/ml), aztreonam (48 microg/ml), piperacillin-tazobactam (192-4 microg/ml), piperacillin (192 microg/ml), and the above three drug combination had bacteriostatic effects on all four strains. Colistin, three time breakpoint of aztreonam, piperacillin, or piperacillin-tazobactam, and the three drug combination of aztreonam, ceftazidime, and amikacin were effective in vitro.

Microbial contamination of fruit and vegetables and their disinfection.

We evaluated the microbial contamination of 17 types of vegetable and 10 types of fruit after 30-s washing with tap water with and without subsequent disinfection by 10-min immersion in 0.01% (100 ppm) sodium hypochlorite. The mean microbial contamination level of 9 types of leafy vegetable was 2.8 x 10(5) colony-forming units (CFU)/g after washing with water and 3.4 x 10(4) CFU/g after washing followed by disinfection. The mean microbial contamination level of 8 types of nonleafy vegetable was 3.4 x 10(4) CFU/g after washing with water and 1.0 x 10(4) CFU/g after washing followed by disinfection. The mean microbial contamination level of 10 types of unpeeled fleshy fruit was 9.3 x 10(3) CFU/g after washing with water and 1.3 x 10(3) CFU/g after washing followed by disinfection. The contaminants in vegetables and unpeeled fruit were similar after washing and after washing followed by disinfection, including Pseudomonas fluorescens and Pseudomonas aeruginosa. The contamination did not markedly decrease even after disinfection with sodium hypochlorite. However, the flesh of each type of peeled fruit showed no or only low levels of contamination (

Microbiological safety of bottled mineral water in patients susceptible to infections.

We evaluated the microbiological safety of bottled mineral water products commercially available in Japan. Of 10 bottled mineral water products manufactured in Japan, no bacteria or fungi were detected in 9 (90%), but 1 (10%) contained 1.8x10(3) colony-forming units (cfu)/mL. Of 12 bottled mineral water products manufactured in the EU, 11 (91.7%) contained 23-3.5x10(4) cfu/mL. On the other hand, of 5 bottled mineral water products manufactured in North America, 2 (40%) contained 2.3x10(2)-2.5x10(3) cfu/mL. The detected microorganisms were glucose-nonfermentative Gram-negative bacilli such as Brevundimonas vesicularis, Moraxella spp., and Burkholderia cepacia, but Pseudomonas aeruginosa was not detected in any product. For immunocompromised host patients being managed in ultra-clean rooms, the examined bottled mineral water products manufactured in Japan, except 1, were microbiologically safe.