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PubMed2001

A test for cleaning and disinfection processes in a washer-disinfector.

Disinfection processes such as heat, aldehydes or alcohols kill vegetative microorganisms but do not necessarily remove other organic contamination. Organic residues impair the result of low-temperature sterilisation processes. Heat-stable organic residues may give rise to clinical symptoms in the patient. Standards are available in Britain and in Sweden for the examination of cleaning processes in washer-disinfectors. The test substances are artificial soil or blood. These standards are based on visual inspection of instruments or equipment. They cannot be used for examination of tubular instruments, nor can they be quantified. For validation of cleaning procedures a simple quantifiable method, which can be performed in an infection control laboratory is needed. We have used suspensions in horse blood of Enterococcus faecalis bacteria and Bacillus subtilis spores to test disinfection and cleaning in a washer-disinfector. Instruments used for laparoscopic surgery were contaminated with a blood bacteria suspension containing 10(7) organisms/ml and then dried and processed in a washer-disinfector using a regular process. Remaining microbial contamination was cultured quantitatively. Nineteen objects were investigated in 10 experiments each. Cleaning, measured as log reduction >5-6 of B. subtilis, was achieved on surfaces that were adequately in contact with the water flow in the machine. Disinfection (and cleaning) measured as log reduction >5-6 of E. faecalis was successful at all points examined. The test method is simple and quantifiable, and can be used to evaluate and to improve cleaning and disinfection processes.

Animals↗
PubMed2003

Comparison of antiadhesive and antibacterial effects of antiseptics on Streptococcus sanguinis.

Three antiseptic (chlorhexidine, Olaflur, Octenisept) and one putative antiadhesive (chitosan) agent were investigated for their effect on viable planktonic and attached Streptococcus sanguinis cells. The bacterial pretreatment with each chemotherapeutic was performed in two steps: (i) After the exposure of planktonic streptococci to the antiseptics, the cells were suspended in human sterile saliva and allowed to attach to human enamel for 60 min; (ii) After 60 min in the flow chamber system, initially attached streptococci were treated with these agents. The microbial viability was monitored by the percentage of vital streptococci determined by fluorescence microscopy and cell reproduction. In comparison with the negative control NaCl, the non-bactericidal chitosan derivative showed distinctive antiadhesive properties. For both treatment procedures, the efficacy of the antiseptics in reducing the viability of planktonic and attached streptococci was Octenisept > Olaflur > chlorhexidine > saline > chitosan. Further studies appear warranted to develop new antiplaque/antibiofilm strategies involving highly efficient bactericidals with antiadhesive formulations.

Anti-Infective Agents, Local↗
PubMed1996

An evaluation of the Nd:YAG laser in periodontal pocket therapy.

The aim of this study was to determine whether the Nd:YAG laser energies of 50 and 80 mJ at 10 pulses per second (pps) were capable of improving the clinical parameters associated with periodontal disease. These energy settings were chosen as previous work indicated that higher values would damage root surfaces and that 80 mJ had an in-vitro bactericidal effect. Eighty periodontally affected sites in teeth scheduled for extraction from 11 patients with adult periodontitis were randomly placed in one of the following four treatment groups: 1. laser treatment at 50 mJ, 10 pps for 3 minutes; 2. laser treatment at 80 mJ, 10 pps for 3 minutes; 3. scaling and 4. untreated control. Probing depth, bleeding on probing (BOP), plaque index, gingival index and gingival crevicular fluid (GCF) volume were measured at baseline and week 6. Baseline subgingival microbiological samples were collected, then repeated immediately after treatment and at week 6 to assess the total anaerobic colony forming units (CFU). Only the scaling group showed a significant reduction in pocket depth and BOP (P < 0.001). The microbial samples taken immediately after scaling and laser at 80 mJ and 10 pps treatments showed a significant reduction in total CFU compared with the baseline (P < 0.01), which was sustained only in the scaling group until week 6. Electron microscopy did not reveal any heat damage on the root surfaces. This study demonstrated that application of Nd:YAG laser pulses of 50 mJ and 80 mJ failed to improve the clinical and microbiological parameters of periodontal disease.

Adult↗
PubMed1999

A method of profiling microbial communities based on a most-probable-number assay that uses BIOLOG plates and multiple sole carbon sources.

A new approach to the community-level BIOLOG assay was proposed. This assay, which we call the BIOLOG-MPN assay, is a most-probable-number (MPN) assay that uses BIOLOG plates and multiple sole carbon sources, and the profiles obtained by this assay consist of MPNs estimated for the substrates in the BIOLOG plates. In order to demonstrate the performance of the BIOLOG-MPN assay, it was applied to pure cultures, model bacterial communities that contain two strains in different ratios, and microbial community samples. MPN estimation using BIOLOG plates worked well for the substrates on which utilizers can grow at a sufficiently high rate for color development under the conditions of the assay procedure. Furthermore, the results obtained using model communities showed that the MPNs obtained reflected the mixing ratios of pure cultures in the model communities. The profiles obtained using model communities and community samples were differentiated properly by statistical analyses. The results suggest that the BIOLOG-MPN assay is a promising procedure for obtaining a quantitative picture of the community structure.

Carbon↗
PubMed1998

Changes in microbiological and physicochemical properties of bee pollen by application of gamma irradiation and ozone treatment.

The effects of gamma irradiation and ozone treatment on microbiological and physicochemical properties of bee pollen were investigated. Gamma irradiation at 7.5 kGy reduced the total microbial loads below detection levels (>10(2) CFU g(-1)), but after ozone treatment of up to 18 ppm for 8 h the total aerobic bacteria were found in concentrations of more than 10(3) CFU g(-1). Physicochemical properties such as amino acid and fatty acid composition, thiobarbituric acid value, mineral content, and pigment were not significantly changed by gamma irradiation, whereas ozone treatment caused significant changes in fatty acid composition, thiobarbituric acid value, and pigment by lipid oxidation and decoloration (P < 0.05).

Amino Acids↗
PubMed2005

Evaluation of nested PCR assays for the detection of Legionella pneumophila in a wide range of aquatic samples.

AIMS: To compare the sensitivities of two nested PCR assays for the detection of Legionella pneumophila to each other and to the plate counting technique (ISO 11731) in a wide range of aquatic samples. METHODS AND RESULTS: The nested PCR assay with the primer set LEG 225-LEG 858 revealed 56% of the 46 analysed aquatic samples as being positive for Legionella spp., while the primer set JFP-JRP yielded 98% positive samples. The detection was confirmed by sequencing the PCR products. These results are considerably higher than the result obtained with the plate counting technique (41%), indicating the higher sensitivity of PCR-based diagnostic methods. As the PCR assay with the LEG 225-LEG 858 primer set resulted in a lower number of positive samples, it is considered not sensitive enough for aquatic samples. Similar results for the respective primer sets were obtained for the detection of the species L. pneumophila, responsible for 90% of all human Legionella infections, in the aquatic samples analysed. Both microbial community analysis by PCR-denaturing gradient gel electrophoresis and the analysis of biotic and abiotic water quality parameters revealed no relation between L. pneumophila-positive and -negative samples and the physico-chemical and bacteriological characteristics of the aquatic samples. CONCLUSIONS: The results show the additional value of the PCR assay with the JFP-JRP primer set compared with the plate counting technique, as well as its applicability in a wide range of aquatic samples. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows the importance of comparing different primer sets for nested PCR assays for the detection of L. pneumophila in aquatic samples, as well as the lower sensitivity of the widely accepted plate counting technique (ISO 11731).

Base Sequence↗
PubMed2005

Shelf life extension of minimally processed cabbage and cucumber through gamma irradiation.

The influence of irradiation of minimally processed cabbage and cucumber on microbial safety, texture, and sensory quality was investigated. Minimally processed, polyethylene-packed, and irradiated cabbage and cucumber were stored at refrigeration temperature (5 degrees C) for 2 weeks. The firmness values ranged from 3.23 kg (control) to 2.82 kg (3.0-kGy irradiated samples) for cucumbers, with a gradual decrease in firmness with increasing radiation dose (0 to 3 kGy). Cucumbers softened just after irradiation with a dose of 3.0 kGy and after 14 days storage, whereas the texture remained within acceptable limits up to a radiation dose of 2.5 kGy. The radiation treatment had no effect on the appearance scores of cabbage; however, scores decreased from 7.0 to 6.7 during storage. The appearance and flavor scores of cucumbers decreased with increasing radiation dose, and overall acceptability was better after radiation doses of 2.5 and 3.0 kGy. The aerobic plate counts per gram for cabbage increased from 3 to 5 log CFU (control), from 1.85 to 2.93 log CFU (2.5 kGy), and from a few colonies to 2.6 log CFU (3.0 kGy) after 14 days of storage at 5 degrees C. A similar trend was noted for cucumber samples. No coliform bacteria were detected at radiation doses greater than 2.0 kGy in either cabbage or cucumber samples. Total fungal counts per gram of sample were within acceptable limits for cucumbers irradiated at 3.0 kGy, and for cabbage no fungi were detected after 2.0-kGy irradiation. The D-values for Escherichia coli in cucumber and cabbage were 0.19 and 0.17 kGy, and those for Salmonella Paratyphi A were 0.25 and 0.29 kGy for cucumber and cabbage, respectively.

Brassica↗
PubMed2006

Microbial flora on operating room telephones.

Approximately 500,000 surgical site infections (SSIs) occur each year in the United States. The purpose of this study was to determine if the bacteria most frequently involved in SSIs could be found on telephones in the OR. Twenty-six cultures were taken from telephones in 14 ORs and two substerile rooms at a large teaching medical center. Using standard laboratory procedures, the researchers identified coagulase-negative staphylococci in the cultures. The study found that telephones in the OR can serve as reservoirs for SSI-causing bacteria.

Bacterial Infections↗
PubMed2001

Growth and histamine formation of Morganella morganii in determining the safety and quality of inoculated and uninoculated bluefish (Pomatomus saltatrix).

The objective of this study was to determine the effect of normal microflora and Morganella morganii on histamine formation and olfactory acceptability in raw bluefish under controlled storage conditions. Fillets inoculated with and without M. morganii were stored at 5, 10, and 15 degrees C for 7 days. Microbial isolates from surface swabs were identified and screened for histidine decarboxylase activity. Olfactory acceptance was performed by an informal sensory panel. Histamine levels were quantified using high-performance liquid chromatography and fluorescence detection. While olfactory acceptance decreased, histamine concentration and bacterial counts increased. Storage temperature had a significant effect on histamine levels, bacterial counts, and olfactory acceptance of the bluefish. Inoculation with M. morganii had a positive significant effect on histamine formation for bluefish held at 10 and 15 degrees C (P < 0.0001). The results of the study will serve in supporting U.S. Food and Drug Administration (FDA) regulations regarding guidance and hazard levels of histamine in fresh bluefish.

Animals↗
PubMed2001

[Microbiological contamination of intraoperatively collected erythrocyte concentrate in mechanical autotransfusion in tumor surgery].

Intraoperative autotransfusion is an effective method of the autologous haemotherapy. Gamma irradiation of the salvaged erythrocyte concentrates leads to inactivation of malignant cells and allows retransfusion in oncologic surgery. The risk of microbiological contamination of the autologous blood product has not been systematically studied in this field of surgery. During the evaluation of intraoperative autotransfusion in oncologic surgery at the University Hospital of Leipzig, the salvaged blood of 46 patients who underwent tumour surgery (urology, gynaecology and neurosurgery) was processed to a washed erythrocyte concentrate with a Cell Saver 5 or a Haemolite 2 (Haemonetics). Specimens for microbiological studies were taken from the area of surgery, the reservoir bottle of the autotransfusion system and the salvaged erythrocyte concentrates. The investigation with plate cultures and the identification of microorganisms were performed according to standard procedures of microbiological diagnosis. The ethics committee of the University Hospital of Leipzig approved the study protocol. We found a microbiological contamination in 10 of 46 salvaged erythrocyte concentrates (22%), above all the growth of saprophytes (Streptococcus, Staphylococcus and Neisseria species). We also identified bacteria which have their habitats in the intestinal tract (E. coli and Enterococcus). The highest rate of microbial contaminated erythrocyte concentrates was found in urologic oncosurgery (37%). In neurosurgery (laminectomy and craniotomy including tumour extirpation), there was no bacterial growth in the salvaged erythrocyte concentrate. Intraoperative autotransfusion should be avoided during surgery touching the urinary tract, e.g. prostatectomy and cystectomy. In neurosurgery, it seems to be safe as far as microbiological contamination is concerned. To assess the real risk of microbiological contamination of the salvaged erythrocytes, more investigations are necessary to determine not only the extent and nature of contamination but also the clinical effects.

Adult↗
PubMedDate not supplied

Airborne viable particles and total number of airborne particles: comparative studies of active air sampling.

This paper presents results from a comparative study of different impaction air samplers under simulated operational conditions in a controlled environment. Furthermore measurements have been performed in a body-box during evaluation of clean-room clothing and in a day care center. Microbial monitoring of controlled environments can be performed in several ways, and with the aid of different sampling instruments. Knowing the limitations of the chosen system is of vital importance for the correct evaluation and interpretation of the results. The number of Colony Forming Units (CFU) detected by one method cannot be directly compared with results from another method. The obtained results seem to indicate that in a controlled environment the exposure situation with a human contamination source gives an approximate relationship between the total number of airborne particles and the number of aerobic airborne viable particles detectable with standard methods.

Air Microbiology↗
PubMed1990

[The problems of microbial indication of water quality].

In three experimental series the reliability of contemporary microbial indicators has been verified. It was shown that about 30% colonies growing on the Endomedium belongs not to family Enterobacteriaceae. The share of the so-called faecal coliforms growing at 44 degrees C is 54.6%. In comparison with the initial cultivations at the second passage this part increasing at about 126%. In waters of various quality is the viability of E. coli the smallest. Also on the Mc Conkey medium at 44 degrees C disappeared the colonies in order: E. coli (3-14 days), K. pneumoniae (3-21 days) C. freundii (21-28 days). This results brings an evidence that the existing indicator system for water quality should be revised.

Citrobacter↗
PubMed2005

Effect of sanitizing treatments on removal of bacteria from cantaloupe surface, and re-contamination with Salmonella.

There are many reports of disease due to consumption of cantaloupes contaminated at the surface with enteric pathogens. Salmonella is among the most frequently reported cause of foodborne outbreaks of gastroenteritis in the United States. Research was undertaken to determine the effects of sanitizer and hot water treatments on microbial populations on cantaloupe surfaces and to determine whether prior decontamination of melons by sanitizer treatment affects vulnerability to recontamination by Salmonella. Cantaloupes were sanitized with 200 ppm chlorine or 2.5% hydrogen peroxide solution for 2 min, or hot water (96 degrees C) for 2 min and were held at 5 degrees C for 24 h. Hot water treatments reduced the microbial populations on cantaloupe surface by 4.9 log reduction while H2O2 or chlorine caused approximately 2.6 log unit reduction on cantaloupe surfaces. When sanitized or hot water treated whole cantaloupes were re-inoculated with Salmonella. Higher populations of Salmonella were recovered from sanitized cantaloupes than from the untreated controls; recovery was greater from hot water treated cantaloupes than from cantaloupes treated with chlorine or hydrogen peroxide. The results of this study clearly show that sanitized cantaloupes are susceptible to recontamination if exposed to a human bacterial pathogen during subsequent handling.

Anti-Infective Agents, Local↗
PubMed2000

Staphylococcus aureus causes acute inflammatory episodes in the cornea during contact lens wear.

The aim of this investigation was to determine the ability of Staphylococcus aureus to cause keratitis during contact lens wear in the rabbit. Rabbits were fitted with hydrogel lenses and wore them on an extended wear schedule for 7 weeks. At the end of each week of wear, one drop of S. aureus was added to the eyes, and the eyes were monitored using slit lamp bio-microscopy, with subsequent microbial and histological studies of the cornea. In the presence of S. aureus, keratitis occurred after 3 weeks of lens wear, a condition that was characterized by diffuse infiltration of the peripheral cornea and limbal redness. In the absence of bacteria, there was no infiltration until week 6. Contact lens wear also prolonged the retention of bacteria in the eye. Extended contact lens wear and S. aureus caused non-ulcerative keratitis in a rabbit model, and disturbances in the ability of the eye to remove bacteria.

Acute Disease↗
PubMed1991

A comparison of human and animal mouth flora.

Human bite injuries are often believed to be more serious than animal bite injuries. One reason for this distinction might be a species difference in the microbial mouth flora. In order to compare this possibility, we cultured the mouths of humans, dogs, and cats. Human oral flora contained the smallest number of bacteria followed by dog and cat oral flora, respectively. Veillonella parvula was cultured from all humans and was also found in dog and cat mouths, but less frequently. V parvula is not usually considered a virulent organism but occasionally may become a pathogen and should not be overlooked especially following human bite injury.

Animals↗
PubMed2003

Synergetic effect of gamma irradiation and moisture content on decontamination of sewage sludge.

Samples of concentrated municipal sewage sludge, stored for 2, 4 and 6 months, with moisture contents of 2%, 20%, 40%, 60%, and 80% were exposed to doses of 0, 1, 2, 3, 4 and 5 kilogray (kGy) of gamma irradiation. Immediately after irradiation, total microbial count and bacterial pathogens in sewage sludge were determined. The results indicated that in all tested sewage sludge samples, bacterial pathogens including Enterobacter sp., Klebsiella sp., Salmonella sp., and Escherichia coli were initially detected. All doses of gamma irradiation reduced the total counts of microorganisms. D(10) of total count decreased with increase in the moisture content of the sewage sludge. The lowest lethal dose for tested bacterial pathogens was 5 kGy in air dried sewage sludge. In addition for wet sewage sludge having more than 40% moisture, the lethal dose was 1 kGy, for samples taken at different storage periods 2, 4 and 6 months, and therefore the cost per unit could be decreased to half when wet sewage sludge (about 50% moisture) was used.

Analysis of Variance↗
PubMed1990

The survival and transfer of microbial contamination via cloths, hands and utensils.

Survival and transfer of bacteria from laminated surfaces and cleaning cloths were investigated under laboratory conditions. Drying produced substantial reductions in numbers of recoverable organisms and achieved satisfactory decontamination of clean laminate surfaces. On soiled surfaces and on clean and soiled cloths, Gram-positive and some Gram-negative species survived for up to 4 h, and in some cases up to 24 h. Where contaminated surfaces or cloths came into contact with the fingers, a stainless steel bowl, or a clean laminate surface, organisms were transferred in sufficient numbers to represent a potential hazard if in contact with food.

Bacteria↗
PubMed1997

The relationship between microbial factors and gingival crevicular fluid glycosaminoglycans in human adult periodontitis.

Counts of cultivable Porphyromonas gingivalis, assays of microbial proteases and the concentration in gingival crevicular fluid of proteoglycan metabolites were investigated at periodontitis and gingivitis sites in 16 patients with chronic adult periodontitis before and after treatment. Two periodontitis sites and two gingivitis sites were selected from each patient on the basis of a clinical examination. Gingival crevicular fluid from each site was analysed for the concentrations of the glycosaminoglycans chondroitin-4-sulphate and hyaluronan and subgingival plaque samples were analysed for cultivable P. gingivalis and microbial trypsin-like proteases assayed by benzoyl-DL-arginine-naphthylamide (BANA) hydrolysis. Significantly higher concentrations (p = 0.007) of chondroitin-4-sulphate were found at periodontitis than gingivitis sites but there was no significant difference in hyaluronan (p = 0.36) between these sites. Although the majority of periodontal sites were P. gingivalis-negative (23/32), there were significantly higher concentrations of chondroitin-4-sulphate (p = 0.05) and hyaluronan (p = 0.04) at the P. gingivalis-positive, compared to negative, periodontitis sites. At BANA-positive periodontitis sites there were also higher concentrations of chondroitin-4-sulphate (p = 0.0015) and hyaluronan (p = 0.0001) than at BANA-positive gingivitis sites. There was a significant decrease in concentrations of chondroitin-4-sulphate and hyaluronan at periodontitis sites after treatment. This study lends support to the hypothesis that P. gingivalis may be actively involved in the destruction of connective tissue components at culture-positive sites but shows that elevated concentrations of connective tissue breakdown products may occur in gingival crevicular fluid from periodontal sites where this organism is absent.

Adult↗