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Neonatal thermal care, part II: Microbial growth under temperature probe covers.

PURPOSE: To determine if temperature probe covers contribute to nosocomial infections by providing an environment for skin microbe colonization. DESIGN: Descriptive, comparative design with infants randomized into two groups: foam probe cover and hydrogel probe cover. Skin cultures were obtained 72 hours after cover placement. Bacterial growth was quantified after 24 and 48 hours. Skin integrity was assessed using visual irritations scores (VIS) every 5 minutes for 30 minutes following cover removal. ANOVA and ANOVA-RM were used to compare amounts of microbes and VIS scores between groups. SAMPLE: Twenty-six medically stable infants, 29 to 34 weeks gestational age, less than 10 days postbirth, and in an incubator. MAIN OUTCOME VARIABLE: Microbial growth and VIS. RESULTS: There were no statistically significant differences in microbial growth and VIS scores between groups (p > .05). Clinical significance was noted in VIS scores. Infants who had foam covers had more sustained irritation scores.

Analysis of Variance↗

Thermal properties and granulometry of dried powders strongly influence the effectiveness of heat treatment for microbial destruction.

The thermal treatment of Saccharomyces cerevisiae cells, which were homogeneously incorporated into dried wheat flour particles, was performed for various particle radii (0.8 to 1.6, 1.6 to 2.8, 2.8 to 3.2, and 5 mm) and for an initial water activity of 0.20. A new high-temperature short-time process developed by our laboratory for powder decontamination was used at 150, 200, and 250 degrees C for 5 to 30 s, and significant destruction of up to a 6.7-log reduction, depending on treatment conditions and granule size, was achieved. This study confirms the strong influence of granulometry on the microbial destruction of homogeneously contaminated powdered products. Moreover, a thermal model was developed that takes into account the thermal properties of each component, the variations during heat treatment, and the energy required for phase change. This model provides a tool for predicting yeast destruction.

Colony Count, Microbial↗

Influence of punctures, cuts, and surface morphologies of golden delicious apples on penetration and growth of Escherichia coli O157:H7.

The ability of Escherichia coli O157:H7 to penetrate and grow within punctures, fresh-cut surfaces, and calyces of Golden Delicious apples was investigated. A three-strain cocktail of E. coli O157:H7 resistant to ampicillin was used to inoculate fresh and 48-h-old punctures, fresh-cut surfaces, and open or closed calyces. A concentric cutting procedure was used to evaluate depth of penetration within punctures and prevent cross contamination during sampling. Within 2 h, E. coli O157:H7 penetrated vertically through the fresh punctures and 3.4 mm within the underlying parenchyma. After 48 h, E. coli O157: H7 cells penetrated up to 5.5 mm within the punctures and >2.6 mm horizontally away from fresh punctures. However, 48-h-old punctures did not permit penetration beyond their boundaries. Fresh-cut surfaces permitted up to 2.8 mm penetration after 24 h. Onset of growth of E. coli O157:H7 occurred 4 to 8 h postinoculation on fresh punctures and fresh-cut surfaces with populations increasing by 3 logs after 48 h. E. coli O157:H7 penetrated within calyces regardless of the extent of opening or method of inoculation. However, E. coli O157:H7 was never recovered from the inner core of apples. Computed tomography scan imaging revealed that closed calyces effectively prevented penetration of sodium iodide solutions within the calyx cavity. Lack of solution penetration may explain why sanitizing treatments are ineffective in inactivating microbial cells within the calyx. Understanding the role of morphological differences in permitting or restricting bacterial penetration may lead to development of more effective strategies to enhance the safety of fresh horticultural products.

Bacterial Adhesion↗

Estimating microbial population counts by 'most probable number' using Microsoft Excel.

A computer-assisted method for determining population counts using the 'most probable number' (MPN) was developed. The Microsoft Excel spreadsheet and its Solver tool were used to generate MPNs, error estimates and confidence limits. Our method was flexible, allowing the use of unbalanced replication schemes and varying replication numbers and inoculation volumes. Furthermore, it required no programming skills and generated fast results, which were comparable to those of standard MPN tables and MPN software.

Colony Count, Microbial↗

Peracetic acid as an alternative wastewater disinfectant to chlorine dioxide.

AIMS: The aim of this study was to compare the efficiency of peracetic acid with that of chlorine dioxide in the disinfection of wastewater from a sewage treatment plant (serving about 650 000 inhabitants) that has been using peracetic acid as a disinfectant since 1998. METHODS AND RESULTS: A total of 23 samplings were made, each consisting of three samples: from secondary effluent, effluent disinfected with 2 mg l(-1) of peracetic acid and effluent disinfected with 2.2 mg l(-1) of chlorine dioxide (contact time 20 min). For each sample, measurements were made of the heterotrophic plate count at 36 degrees C, total and faecal coliforms, Escherichia coli, enterococci, pH, suspended solids and chemical oxygen demand (COD). During the first phase of the experiment the peracetic acid was seen to be less efficient than chlorine dioxide. To improve the disinfectant action a system of mechanical agitation was added which led to a greater efficiency in the inactivation of bacteria of faecal origin. CONCLUSIONS: Both products were found to be influenced by the level of microbial contamination, the amount of suspended solids and COD but not by the pH of the effluent before disinfection. The immediate mixing of the wastewater and disinfectant caused a greater reduction in enterococci. SIGNIFICANCE AND IMPACT OF THE STUDY: Since peracetic acid was seen to produce a high abatement of micro-organisms, it can be considered as a valid alternative to chlorine dioxide in the disinfection of wastewaters.

Bacteria↗

Direct differential counting compared with testing of isolates as means of assessing hydrolytic capacities of natural microbial populations.

The abundance of microorganisms with characteristic, physiological activities in aquatic sediment communities can be estimated either by direct differential counting on selective media or by making isolations followed by testing of the isolates. The two methods have been compared with regard to microorganisms performing hydrolyses of different compounds. The first method was used to examine the effect of a model ecocontaminant on the hydrolytic activities of the sediment population.

Colony Count, Microbial↗

Distribution of viral abundance in the reef environment of Key Largo, Florida.

The distribution of viral and microbial abundance in the Key Largo, Fla., reef environment was measured. Viral abundance was measured by transmission electron microscope direct counts and plaque titer on specific bacterial hosts in water and sediment samples from Florida Bay (Blackwater Sound) and along a transect from Key Largo to the outer edge of the reef tract in Key Largo Sanctuary. Water column viral direct counts were highest in Blackwater Sound of Florida Bay (1.2 x 10(7) viruses per ml), decreased to the shelf break (1.7 x 10(6) viruses per ml), and were inversely correlated with salinity (r = -0.97). Viral direct counts in sediment samples ranged from 1.35 x 10(8) to 5.3 x 10(8)/cm(3) of sediment and averaged nearly 2 orders of magnitude greater than counts in the water column. Viral direct counts (both sediment and water column measurements) exceeded plaque titers on marine bacterial hosts (Vibrio natriegens and others) by 7 to 8 orders of magnitude. Water column viral abundance did not correlate with bacterial direct counts or chlorophyll a measurements, and sediment viral parameters did not correlate with water column microbial, viral, or salinity data. Coliphage, which are indicators of fecal pollution, were detected in two water column samples and most sediment samples, yet their concentrations were relatively low (<2 to 15/liter for water column samples, and <2 to 108/cm(3) of sediment). Our findings indicate that viruses are abundant in the Key Largo environment, particularly on the Florida Bay side of Key Largo, and that processes governing their distribution in the water column (i.e., salinity and freshwater input) are independent of those governing their distribution in the sediment environment.

Animals↗

Local antiseptic therapy for non-gonococcal urethritis: phase II study with Instillagel.

Non-gonococcal urethritis (NGU) is conventionally treated with oral antibiotics. With this Phase II study, we investigated the action of a locally disinfecting substance, Instillagel, in symptomatic NGU. Instillation treatment was performed twice daily to 32 male patients with symptomatic NGU. To evaluate the therapeutic outcome, a smear was taken from the urethra and an urine examination was performed at baseline as well as at 5 and 8 days after the start of the treatment. Pain and micturition symptoms were determined by a questionnaire with analog scales taken before and after therapy. A pre/post comparison of the urethral smears of the patients with symptomatic NGU showed a significant difference (P < 0.0001). The microbial count in the urine did not show a significant difference. The symptoms micturition (P<0.0001) and pain in the urethra (P<0.0001) were significantly improved. This Phase II study confirmed that local antiseptic treatment of NGU can offer an alternative to systemic antibiotic treatment.

Anti-Infective Agents, Local↗

Bacteriological water quality status of River Yamuna in Delhi.

Bacteriological water quality status in terms of total coliform and faecal coliform count was studied on both--east and west banks of river Yamuna in Delhi. Membrane filtration technique was adopted for enumeration of total coliform and faecal coliform count in the river water sample collected on monthly basis for 2 years--2002 and 2003. The study reveals the impact of diverse anthropogenic activities as well as the monsoon effect on the bacterial population of river Yamuna in Delhi stretch. Microbial population contributed mainly through human activities prevailed in the entire stretch of Yamuna river with reduction in bacterial counts during monsoon period due to flushing effect. Bacteriological assessment does not provide an integrated effect of pollution but only indicate the water quality at the time of sampling. Hence, this parameter is time and space specific.

Bacteriological Techniques↗

Indoor air climate and microbiological airborne: contamination in various hospital areas.

Indoor climate indices and microbiological airborne contamination were evaluated in a department of a general hospital in Bologna only partially equipped with an air conditioning system. To determine the environmental parameters, an ANADATA (LSI) climate analyzer with relative transducers was used. The Effective Temperature (ET), the New Effective Temperature (ET*) and the Fanger indices (PMV-PPD) were calculated using the parameters measured. Microbial count measurements were taken with an S.A.S. (Surface Air System) sampler, to ascertain the total bacterial count at 37 degrees C, and the fungal particle, Staphylococcus aureus and Pseudomonas aeruginosa counts. Carbon dioxide air concentrations were also measured to evaluate the efficacy of air exchange. The Fanger indices were not within the range of thermal comfort in most rooms (52% in winter, 62% in summer). Air microbial counts were higher in the hospital wards and surgeries than in the offices and laboratories. In particular, coagulase-positive staphylococci were present only in the air of the patients' rooms. The microbial contamination was not correlated with the air conditioning system, but probably caused by the turnover in the hospital population, the number of people and their behaviour. However the most important measure to prevent airborne contamination and to reduce the number of microorganisms in the air is an efficient source control. Better management of the air conditioning system, by means of adequate air exchange and thermal adjustment, would lead to a notable improvement in indoor air quality, especially in units with hospitalized patients.

Air Conditioning↗

An in vitro oral biofilm model for comparing the efficacy of antimicrobial mouthrinses.

The ability of commercial mouthrinses to reduce total viable counts of mixed microbial populations was examined using a previously developed in vitro model of supragingival plaque. Exploratory experiments aimed at fine-tuning the model indicated that optimal correspondence between in vitro and clinical results for chlorhexidine-containing formulations were obtained at a saliva:medium ratio of 70:30 (v/v); moreover, expanding the microbial population from 5 bacterial species to 5 bacterial species + Candida albicans had no noticeable impact on overall results. The efficacies of 12 different mouthrinse proprietary products containing chlorhexidine, hexetidine, octenidine, Triclosan, plant extracts, or aminefluoride/stannous fluoride vis-à-vis biofilm clearance were compared. All mouthrinses promoted a statistically significant reduction in microbial load compared to distilled water. The herbal- and phenolic-based products were substantially less effective than most chlorhexidine-containing mouthrinses, or mouthrinses containing hexetidine or octenidine. No significant difference between the plaque-clearing plaque-clearing abilities of Listerine and Meridol was observed. This polyspecies biofilm model can be a valuable tool for preclinical testing of antiplaque formulations, particularly during the product development stage.

Anti-Infective Agents, Local↗

Sabella spallanzanii filter-feeding on bacterial community: ecological implications and applications.

The filtration process of Sabella spallanzanii Gmelin on bacterial community was studied in a coastal area of the Northern Ionian Sea (Mediterranean Sea) at three sites, S. Vito, Lido Gandoli and Lido Silvana, where some specimens of S. spallanzanii were transplanted. Analyses were performed both on water and worm samples. A total of six microbial groups were examined: culturable heterotrophic bacteria, total culturable bacteria at 37 degrees C, culturable vibrios, total and fecal coliforms and fecal streptococci. The bacterial densities were usually orders of magnitude higher in the worm homogenates than in the corresponding seawater and the highest values were observed in August. The ability of S. spallanzanii to accumulate the microbial pollution indicators suggests this species can be employed as a bioindicator for monitoring water quality. Moreover, the accumulation capability of S. spallanzanii for specific micro-organisms provides a potential role in sewage bioremediation.

Analysis of Variance↗

Utilisation of a packed-bed biofilm reactor for the determination of the potential of biofilm accumulation in water systems.

An experimental system has been developed that allows the monitoring of biofilm development on supports exposed to water of different characteristics. The system consists of a series of packed-bed reactors filled with glass beads, and by periodically removing biofilm attached to these beads for off-line analyses this provides a means for monitoring biofilm development. Despite its reduced dimensions (6.9 cm long and 1.58 cm in diameter), the experimental system used has a sampling surface of 90.3 cm2 (including only the surface of the glass beads). This allows reproducible and representative samples to be taken from different water systems, providing a reliable and economic method for evaluating in situ the formation of biofilms from different environments. The set-up of the entire experimental system was constructed to meet the demands of field experiments in a well-defined hydrodynamic environment and to allow easy removal of samples for biomass quantification and microscopic observation. Data obtained using this device can be used as an indicator of the risk of biofilm formation in different water systems. This indicator, "the biofilm accumulation potential", represents an effective and representative tool for the monitoring of biofilm development in an integrated antifouling strategy, in order to help keep biofouling, scaling and microbial risks under control. According to the experiments with the packed-bed reactors used with a high flow regime, the ratio TCN/HPC could provide an indication of the state of the biofilm, and lower ratios could indicate a higher biofilm accumulation potential.

Biofilms↗

Effect of Pseudomonas aeruginosa concentration in experimental contact lens-related microbial keratitis.

Pseudomonas aeruginosa adherence in vitro to perfilcon A (ionic, 71% H2O) extended wear soft contact lenses--both new and after 7 days of continuous wear on closed rabbit eyes--was found to be related directly to the bacterial concentration in the contaminating solution. Thirty rabbits wore perfilcon A lenses for 7 days with complete lid closure to mimic contact lens overwear. After 7 days, conjunctival cultures showed no growth of pathogens, but all corneas had developed epithelial cell exfoliation and/or epithelial defects and stromal edema. The lenses were then incubated in various concentrations (10(7), 10(6), 10(5), 10(4), and 10(2) colony-forming units per milliliter or saline control; n = 5/group) of P. aeruginosa suspensions and replaced on their respective corneas with tarsorrhaphies for an additional 48 h. By day 9, corneal thickness had increased significantly, and P. aeruginosa keratitis had developed in 13 of 25 bacterially exposed eyes but not in 5 control eyes. Although with decreasing P. aeruginosa concentration the prevalence of ulcerative microbial keratitis also decreased, the initial concentration of bacteria or the initial extent of soft contact lens-induced corneal damage had no influence on the ultimate clinical severity of the disease.

Animals↗

Determination of the in vivo pharmacodynamic profile of cefepime against extended-spectrum-beta-lactamase-producing Escherichia coli at various inocula.

Cefepime was evaluated in vivo against two inoculum sizes of four strains of Escherichia coli that produced extended-spectrum beta-lactamases (ESBLs) in a murine neutropenic thigh infection model to characterize the pharmacodynamic activity of cefepime in the presence of ESBL-producing bacteria and to evaluate if differences in lengths of cefepime exposure are required with various inocula. Three strains possessed a single enzyme each: TEM-10, TEM-12, and TEM-26. The fourth strain possessed two TEM-derived ESBLs and a third uncharacterized enzyme. Two non-ESBL-producing E. coli strains were included for comparison. Mice received various doses of cefepime to achieve a spectrum of percentages of time the drug was above the MIC (%T>MICs) for each isolate at both inocula. No significant difference in cefepime exposure was required to achieve similar bactericidal effects for ESBL- and non-ESBL-producing isolates when the starting inoculum was 10(5) CFU of E. coli per thigh. The increased MICs observed in vitro for the ESBL-producing strains at 10(7) CFU/ml did not predict the amount of exposure required to achieve a comparable level of bactericidal activity in vivo at the corresponding starting inoculum of 10(7) CFU/thigh. Compared to the cefepime exposure in tests with the lower inoculum (10(5) CFU/thigh), less exposure was required when the starting inoculum was 10(7) CFU/thigh (%T>MIC, 6% versus 26%), such that similar doses (in milligrams per kilogram of body weight) produced similar bactericidal effects with both inocula of ESBL-producing isolates. Equivalent exposures of cefepime produced similar effects against the microorganisms regardless of the presence of ESBL production. Pharmacodynamic profiling undertaken with conventional cefepime MIC determinations predicted in vivo microbial outcomes at both inoculum sizes for the ESBL-producing isolates evaluated in this study. These data support the use of conventional MIC determinations in the pharmacodynamic assessment of cefepime.

Animals↗

Nitrification and autotrophic nitrifying bacteria in a hydrocarbon-polluted soil.

In vitro ammonia-oxidizing bacteria are capable of oxidizing hydrocarbons incompletely. This transformation is accompanied by competitive inhibition of ammonia monooxygenase, the first key enzyme in nitrification. The effect of hydrocarbon pollution on soil nitrification was examined in situ. In a microcosm study, adding diesel fuel hydrocarbon to an uncontaminated soil (agricultural unfertilized soil) treated with ammonium sulfate dramatically reduced the amount of KCl-extractable nitrate but stimulated ammonium consumption. In a soil with long history of pollution that was treated with ammonium sulfate, 90% of the ammonium was transformed into nitrate after 3 weeks of incubation. Nitrate production was twofold higher in the contaminated soil than in the agricultural soil to which hydrocarbon was not added. To assess if ammonia-oxidizing bacteria acquired resistance to inhibition by hydrocarbon, the contaminated soil was reexposed to diesel fuel. Ammonium consumption was not affected, but nitrate production was 30% lower than nitrate production in the absence of hydrocarbon. The apparent reduction in nitrification resulted from immobilization of ammonium by hydrocarbon-stimulated microbial activity. These results indicated that the hydrocarbon inhibited nitrification in the noncontaminated soil (agricultural soil) and that ammonia-oxidizing bacteria in the polluted soil acquired resistance to inhibition by the hydrocarbon, possibly by increasing the affinity of nitrifying bacteria for ammonium in the soil.

Colony Count, Microbial↗

Quantity and characteristics of microorganisms associated with ruminal fluid or particles.

An experiment was conducted to 1) determine quantity of microbial CP and DM associated with undigested feed particles, 2) quantify the amount of microorganisms removed from ruminally incubated forages and 3) compare forage disappearances (D) corrected for microbial contamination, using diaminopimelic acid (DAPA) ratios obtained from particle-associated or fluid-associated microorganisms. Samples of alfalfa hay, bermudagrass hay and orchardgrass hay placed in dacron bags were incubated via ruminal cannula for 6 and 12 h. Whole ruminal contents (WRC) were sampled at the time of bag removal and partitioned into fluid- and particle-associated microorganisms. Particle-associated microorganisms were further divided into loosely-associated (LA) and firmly associated (FA) microorganisms. Percentage microbial CP and DAPA, quantities of OM, CP (mg) and DAPA (micrograms), microbial contamination and microbial contribution to ruminal contents were not affected by time (P greater than .05). The highest concentrations of CP and DAPA were found in fluid and LA, respectively (P less than .01). The total amount (content) of OM, CP and DAPA were highest in FA and lowest in microbes in the fluid fraction. Firmly-associated microorganisms constituted 54.1% of the particle-associated population. Corrected DMD were higher (P less than .01) based on FA than on fluid and LA microbe compositions. Corrected CPD were similar (P greater than .10) between populations except at 6 h, when LA corrected CPD were lower. Theoretically, using FA microorganisms should give the most accurate correction for microbial contamination. However, results indicate that, although the three populations differed in composition, fluid and FA corrected CPD were similar. Corrections based on composition of any of these three populations will yield higher estimates of disappearance compared with uncorrected values.

Animal Feed↗

Effective cleaning and sanitizing of polysulfone ultrafiltration membrane systems.

Polysulfone UF membranes that were soiled by Cheddar cheese whey were successfully cleaned in place. This cleaning procedure was completed in about 1 h. Most cleaning chemicals used were common and inexpensive. The cleaning procedure consisted of rinsing the membrane system for 2 min with water initially and after each cleaning solution. Sodium hydroxide at pH 11.0, with .1% of a nonionic surfactant added, was circulated for 20 min. After a 2-min rinse with water, a 1:1 mixture of nitric and phosphoric acids at pH 2 was circulated for 20 min and rinsed again with water. Finally, sodium hydroxide at pH 11.0, with 200 ppm of sodium hypochlorite added, was circulated for 20 min and rinsed. All cleaning solutions and all rinse waters were at 54 degrees C. Membranes cleaned by this procedure were found to be free from whey residue under examination by scanning electron microscopy. The cleaning process did not damage the membranes even when it was used continuously for 300 h. Microbial populations on the membrane were estimated by incubating small (4-cm2) sections of membrane in screw-cap vials filled with trypticase soy broth. From the portion of vials showing growth after 72 h at 32 degrees C, a most probable microbial population was calculated. Santizing cleaned polysulfone UF membranes with 100 ppm of sodium hypochlorite or 100 ppm of dichloroisocyanurate at 54 degrees C resulted in membranes free from viable microorganisms. When dichloroisocyanurate was used at 10 degrees C and 200 ppm, a most probable microbial population of 290/m2 was found. No microbial growth was detected when cleaned and sanitized membranes were stored in tap water for 24 h. This technique for cleaning UF membranes does not require the use of a holding solution containing santizers to control the growth of residual microorganisms.

Bacteria↗