In vitro modeling of dental water line contamination and decontamination

The contamination of dental unit water lines (DUWL) is an emerging concern in dentistry. The aim of this study was to use an in vitro DUWL to model microbial contamination and evaluate the decontamination efficacy of tetraacetylethylenediamine (TAED) solutions. A DUWL biofilm model used to simulate clinical conditions was used to generate a range of biofilms in DUWL. Three distinct biofilms were generated: (1) biofilm from water, (2) biofilm from a mix of water + contaminating human commensal bacteria, (3) biofilm from water with contaminating oral bacteria added after biofilm formed. The contaminating oral species used were Streptococcus oralis, Enterococcus faecalis and Staphylococcus aureus. Decontamination by simple water flushing or flushing with TAED was evaluated (2, 5 and 10 min intervals). The DUWL tubes were split and samples were plated onto a range of media, incubated and bacteria enumerated. Water flushing did not reduce the number of microorganisms detected. Bacteria were not detected from any of the TAED sampling points for any of the biofilm types tested. Interestingly, if contamination was introduced to new DUWL along with the waterborne species a biofilm was formed containing only the waterborne species. If however, an existing biofilm was present before the introduction of "contaminating" bacteria then these could be detected in the biofilm. This implies that if the DUWL are new or satisfactorily cleaned on a regular basis then the associated cross-contamination aspects are reduced. In conclusion, TAED provides effective control for DUWL biofilms.     

Biofilm problems in dental unit water systems and its practical control

Dental chair units (DCUs) contain integrated systems that provide the instruments and services for a wide range of dental procedures. DCUs use water to cool and irrigate DCU-supplied instruments and tooth surfaces during dental treatment. Water is supplied to these instruments by a network of interconnected narrow-bore (2–3 mm) plastic tubes called dental unit waterlines (DUWLs). Many studies over the last 40 years demonstrated that DUWL output water is often contaminated with high densities of micro-organisms, predominantly Gram-negative aerobic heterotropic environmental bacteria, including Legionella and Pseudomonas species. Untreated DUWLs host biofilms that permit micro-organisms to multiply and disperse through the water network and which are aerosolized by DCU instrument use, thus exposing patients and staff to these micro-organisms, to fragments of biofilm and bacterial endotoxins. This review concentrates on how practical developments and innovations in specific areas can contribute to effective DUWL biofilm control. These include the use of effective DUWL treatment agents, improvements to DCU supply water quality, DCU design changes, development of automated DUWL treatment procedures that are effective at controlling biofilm in the long-term and require minimal human intervention, are safe for patients and staff, and which do not cause deterioration of DCU components following prolonged use.     

Effect of Biocides on Biofilm Bacteria from Dental Unit Water Lines

Microbial biofilm formation in dental unit water lines (DUWL) is a phenomenon that has been recognized for nearly four decades. Water delivered by DUWL can harbor high numbers of bacteria, including opportunistic pathogens. Biofilms on tubing within DUWL may serve as a reservoir for these microorganisms and should therefore be controlled. In this study, the effects of eight biocides were monitored on DUWL biofilms individually and in combination by epifluorescence microscopy and total viable counts (TVC). The effects of sodium dodecyl sulphate (SDS), hydrogen peroxide (H₂O₂), sodium hypochlorite (NaOCl), phenol (Phe), Tween 20 (Tw 20), ethylenediaminetetraacetic acid (EDTA), chlorohexidine gluconate (CHX), and povidine iodine (PI) were tested on DUWL biofilms alone and in combination. PI was found to have negligible effects on biofilm removal either applied alone or in combined form with CHX. Applying all biocides simultaneously did not completely eliminate viable bacteria nor did they remove biofilm. Overall, when combined, the biocides performed better than singly applied products. The most effective biocides were NaOCl and Phe (both alone and in combination).     

Control of microbial contamination in dental unit water systems using tetra-sodium EDTA

Aim:  To examine the efficacy of tetra-sodium EDTA in controlling microbial contamination of dental unit water systems (DUWS).
Methods and Results:  Ten dental units were treated once a week with either 4% or 8% tetra-sodium EDTA for four or two consecutive weeks, respectively. Before treatment, 43% and 60% of the water samples from the air/water triple syringe and high-speed hand-pieces, respectively, exceeded the American Dental Association (ADA) guidelines of 200 CFU ml⁻¹ water during a 6-week baseline period. After each weekend treatment, the levels of microbial contamination in all DUWS fell significantly ( P  < 0·001) to below the ADA guideline. By the end of the week, microbial counts in the outflowing water had returned to baseline levels indicating a transient effect of single doses of tetra-sodium EDTA, and the need for multiple applications. The biofilms were virtually eliminated after a single weekend treatment.
Conclusions:  Tetra-sodium EDTA is effective in controlling microbial contamination in DUWS.
Significance and Impact of the Study:  Inexpensive, effective and safe products for reducing the microbial load of water from DUWS are needed to meet ADA and other national guidelines. Tetra-sodium EDTA can significantly reduce microbial biofilms and bacterial counts in outflowing water, and is compatible for use in DUWS.     

Influence of material and tube size on DUWLs contamination in a pilot plant

Numerous studies have shown that the water discharged from dental unit waterlines (DUWLs) contains high densities of bacteria, especially non-fermenting Gram negative bacteria. The aim of the present study was to investigate how the material (polyethylene-PE and polytetrafluorethylene-PTFE) and size (1.6 and 4.0 mm) of 4 waterlines in a pilot plant influence the level of contamination in the output water. The water contamination was assessed by analyzing the trend of the heterotrophic plate counts at 22 degrees C as a function of time and by testing for non-fermenting Gram negative bacteria. In all waterlines, the bacterial density increased exponentially during the first months and thereafter remained between 10(4) and 10(6) cfu/ml. However, the plate count at 22 degrees C was lower in the water from PTFE tubes and from larger size tubes. Comamonas acidovorans, Pseudomonas aeruginosa and Pseudomonas fluorescens were isolated. Pseudomonas aeruginosa, responsible for infections associated with dental practice, was never isolated in the output water from PTFE tubes. In order to control bacterial contamination the results suggest the use of waterlines made of PTFE on account of their ability to inhibit the colonization and growth of Pseudomonas aeruginosa.     

Aerosolization of mycobacteria and legionellae during dental treatment: low exposure despite dental unit contamination

Dental unit waterlines (DUWL) support growth of a dense microbial population that includes pathogens and hypersensitivity-inducing bacteria, such as Legionella spp. and non-tuberculous mycobacteria (NTM). Dynamic dental instruments connected to DUWL generate aerosols in the work environment, which could allow waterborne pathogens to be aerosolized. The use of the real-time quantitative polymerase chain reaction (qPCR) provides a more accurate estimation of exposure levels compared with the traditional culture approach. Bioaerosol sampling was performed 13 times in an isolated dental treatment room according to a standardized protocol that included four dental prophylaxis treatments. Inhalable dust samples were taken at the breathing zone of both the hygienist and patient and outside the treatment room (control). Total bacteria as well as Legionella spp. and NTM were quantified by qPCR in bioaerosol and DUWL water samples. Dental staff and patients are exposed to bacteria generated during dental treatments (up to 4.3 E + 05 bacteria per m(3) of air). Because DUWL water studied was weakly contaminated by Legionella spp. and NTM, their aerosolization during dental treatment was not significant. As a result, infectious and sensitization risks associated with legionellae and NTM should be minimal.     

Activity of Microorganisms in Acid Mine Water I. Influence of Acid Water on Aerobic Heterotrophs of a Normal Stream

Comparison of microbial content of acid-contaminated and nonacid-contaminated streams from the same geographical area indicated that nonacid streams contained relatively low numbers of acid-tolerant heterotrophic microorganisms. The acid-tolerant aerobes survived when acid entered the stream and actually increased in number to about 2 × 10³ per ml until the pH approached 3.0. The organisms then represented the heterotrophic aerobic microflora of the streams comprised of a mixture of mine drainage and nonacid water. A stream which was entirely acid drainage did not have a similar microflora. Most gram-positive aerobic and anaerobic bacteria died out very rapidly in acidic water, and they comprised a very small percentage of the microbial population of the streams examined. Iron- and sulfur-oxidizing autotrophic bacteria were present wherever mine water entered a stream system. The sulfur-oxidizing bacteria predominated over iron oxidizers. Ecological data from the field were verified by laboratory experiments designed to simulate stream conditions.     

Indigenous and contaminant microbes in ultradeep mines

Rock, air and service water samples were collected for microbial analyses from 3.2 kilometres depth in a working Au mine in the Witwatersrand basin, South Africa. The approximately metre-wide mined zone was comprised of a carbonaceous, quartz, sulphide, uraninite and Au bearing layer, called the Carbon Leader, sandwiched by quartzite and conglomerate. The microbial community in the service water was dominated by mesophilic aerobic and anaerobic, alpha-, beta- and gamma-Proteobacteria with a total biomass concentration approximately 10(4) cells ml(-1), whereas, that of the mine air was dominated by members of the Chlorobi and Bacteroidetes groups and a fungal component. The microorganisms in the Carbon Leader were predominantly mesophilic, aerobic heterotrophic, nitrate reducing and methylotrophic, beta- and gamma-Proteobacteria that were more closely related to service water microorganisms than to air microbes. Rhodamine WT dye and fluorescent microspheres employed as contaminant tracers, however, indicated that service water contamination of most of the rock samples was < 0.01% during acquisition. The microbial contaminants most likely originated from the service water, infiltrated the low permeability rock through and accumulated within mining-induced fractures where they survived for several days before being mined. Combined PLFA and terminal restriction fragment length profile (T-RFLP) analyses suggest that the maximum concentration of indigenous microorganisms in the Carbon Leader was < 10(2) cells g(-1). PLFA, 35S autoradiography and enrichments suggest that the adjacent quartzite was less contaminated and contained approximately 10(3) cells gram(-1) of thermophilic, sulphate reducing bacteria, SRB, some of which are delta-Proteobacteria. Pore water and rock geochemical analyses suggest that these SRB's may have been sustained by sulphate diffusing from the adjacent U-rich, Carbon Leader where it was formed by radiolysis of sulphide.     

Heterogeneity in the efficacy of dental chemical disinfectants on water-derived biofilms in vitro

Abstract

Conditions in dental unit waterlines are favourable for biofilm growth and contamination of dental unit water. The aim of this study was to assess the effect of several chemical disinfectants on bacteria in a biofilm model. Water-derived biofilms were grown in a static biofilm model (Amsterdam Active Attachment model), using two growth media. Biofilms were challenged with Alpron/Bilpron, Anoxyl, Citrisil, Dentosept, Green & Clean, ICX and Oxygenal in shock dose and maintenance doses. The concentration and the composition of the chemical disinfectants influenced the number of culturable bacteria in the biofilms. The application of a single shock dose followed by a low dose of the same chemical disinfectants resulted in the greatest suppression of viable bacteria in the biofilms. Exposure to Citrisil and ICX consistently resulted in failure to control the biofilms, while Alpron/Bilpron had a substantial and relevant effect on the number of bacteria in the biofilms.     

Biocide sensitivity of Vermamoeba vermiformis isolated from dental-unit-waterline systems

This study isolated amoebae from two different dental-unit-waterline (DUWL) sources, a simulated laboratory system (sDUWL) and a decommissioned system (dDUWL), within 24 h of it being dismantled from a working clinical practice. Molecular profiles of the isolates were determined and morphological characteristics of the test organisms were examined using microscopy. DNA barcoding identified the amoebae from both sources as being Vermamoeba vermiformis (previously described as Hartmannella vermiformis). These amoebae have been deposited at the Culture Collection of Algae and Protozoa and as CCAP 1534/16, GenBank accession number KC161965 (in-vitro simulated system) and CCAP 1534/17, GenBank accession number KC188996 (decommissioned system). The organisms from both sources exhibited two main forms: motile trophozoites and non-motile cysts. Mature cysts displayed natural fluorescence with excitation and emission wavelength of 488 nm, attributed to the presence of natural flavins and nicotinamide compounds. Both the encysted and the trophozoite forms of V. vermiformis remained unaffected when exposed to the proprietary biocides tested at the concentrations recommended for use in dental surgeries and cysts successfully excysted, to release trophozoites upon subsequent culture. However, two out of three proprietary dental biocides tested on isolated heterotrophic bacteria were effective at controlling bacterial contamination.     

Establishing a laboratory model of dental unit waterlines bacterial biofilms using a CDC biofilm reactor

In this study, a laboratory model to reproduce dental unit waterline (DUWL) biofilms was developed using a CDC biofilm reactor (CBR). Bacteria obtained from DUWLs were filtered and cultured in Reasoner’s 2A (R2A) for 10 days, and were subsequently stored at ?70°C. This stock was cultivated on R2A in batch mode. After culturing for five days, the bacteria were inoculated into the CBR. Biofilms were grown on polyurethane tubing for four days. Biofilm accumulation and thickness was 1.3 × 10⁵ CFU cm^?2 and 10–14 μm respectively, after four days. Bacteria in the biofilms included cocci and rods of short and medium lengths. In addition, 38 bacterial genera were detected in biofilms. In this study, the suitability and reproducibility of the CBR model for DUWL biofilm formation were demonstrated. The model provides a foundation for the development of bacterial control methods for DUWLs.     

Update on infectious risks associated with dental unit waterlines

Modern dental chair units consist of a network of interconnected narrow-bore plastic tubes called dental unit waterlines (DUWLs). The water delivered by these DUWLs acts as both a coolant for a range of instruments and an irrigant during dental treatments. The quality of water is of considerable importance because both patients and dental team are regularly exposed to water and aerosols generated by dental equipment. Studies have demonstrated that DUWLs provide a favourable environment for microbial proliferation and biofilm formation, and that water is consequently often contaminated with high densities of various microorganisms (bacteria, fungi, protozoa, viruses). The presence of high levels of microbial contamination may be a health problem for dentists and patients, especially those who are immunocompromised. The current status of knowledge on microbial contamination of DUWLs is presented, with an emphasis on the infectious risk associated with DUWLs and on the various approaches for disinfecting and protecting DUWLs.     

Molecular analysis of microbial communities in endotracheal tube biofilms

Background

Ventilator-associated pneumonia is the most prevalent acquired infection of patients on intensive care units and is associated with considerable morbidity and mortality. Evidence suggests that an improved understanding of the composition of the biofilm communities that form on endotracheal tubes may result in the development of improved preventative strategies for ventilator-associated pneumonia.

Methodology/Principal Findings

The aim of this study was to characterise microbial biofilms on the inner luminal surface of extubated endotracheal tubes from ICU patients using PCR and molecular profiling. Twenty-four endotracheal tubes were obtained from twenty mechanically ventilated patients. Denaturing gradient gel electrophoresis (DGGE) profiling of 16S rRNA gene amplicons was used to assess the diversity of the bacterial population, together with species specific PCR of key marker oral microorganisms and a quantitative assessment of culturable aerobic bacteria. Analysis of culturable aerobic bacteria revealed a range of colonisation from no growth to 2.1×10⁸ colony forming units (cfu)/cm² of endotracheal tube (mean 1.4×10⁷ cfu/cm²). PCR targeting of specific bacterial species detected the oral bacteria Streptococcus mutans (n = 5) and Porphyromonas gingivalis (n = 5). DGGE profiling of the endotracheal biofilms revealed complex banding patterns containing between 3 and 22 (mean 6) bands per tube, thus demonstrating the marked complexity of the constituent biofilms. Significant inter-patient diversity was evident. The number of DGGE bands detected was not related to total viable microbial counts or the duration of intubation.

Conclusions/Significance

Molecular profiling using DGGE demonstrated considerable biofilm compositional complexity and inter-patient diversity and provides a rapid method for the further study of biofilm composition in longitudinal and interventional studies. The presence of oral microorganisms in endotracheal tube biofilms suggests that these may be important in biofilm development and may provide a therapeutic target for the prevention of ventilator-associated pneumonia.     

Effect of biofilm in irrigation pipes on microbial quality of irrigation water

Aims: The focus of this work was to investigate the contribution of native Escherichia coli to the microbial quality of irrigation water and to determine the potential for contamination by E. coli associated with heterotrophic biofilms in pipe‐based irrigation water delivery systems. Methods and Results: The aluminium pipes in the sprinkler irrigation system were outfitted with coupons that were extracted before each of the 2‐h long irrigations carried out with weekly intervals. Water from the creek water and sprinklers, residual water from the previous irrigation and biofilms on the coupons were analysed for E. coli. High E. coli concentrations in water remaining in irrigation pipes between irrigation events were indicative of E. coli growth. In two of the four irrigations, the probability of the sample source, (creek vs sprinkler), being a noninfluential factor, was only 0·14, that is, source was an important factor. The population of bacteria associated with the biofilm on pipe walls was estimated to be larger than that in water in pipes in the first three irrigation events and comparable to one in the fourth event. Conclusion: Biofilm‐associated E. coli can affect microbial quality of irrigation water and, therefore, should not be neglected when estimating bacterial mass balances for irrigation systems. Significance and Impact of the Study: This work is the first peer‐reviewed report on the impact of biofilms on microbial quality of irrigation waters. Flushing of the irrigation system may be a useful management practice to decrease the risk of microbial contamination of produce. Because microbial water quality can be substantially modified while water is transported in an irrigation system, it becomes imperative to monitor water quality at fields, rather than just at the intake.     

Preliminary Studies on Membrane Filtration for the Production of Potable Water: A Case of Tshaanda Rural Village in South Africa

Ultrafiltration (UF) systems have been used globally for treating water from resources including rivers, reservoirs, and lakes for the production of potable water in the past decade. UF membranes with a pore size of between 0.1 and 0.01 micrometres provide an effective barrier for bacteria, viruses, suspended particles, and colloids. The use of UF membrane technology in treating groundwater for the supply of potable water in the impoverished and rural village, Tshaanda (i.e., the study area) is demonstrated. The technical and administrative processes that are critical for the successful installation of the pilot plant were developed. Given the rural nature of Tshaanda, the cultural and traditional protocols were observed. Preliminary results of the water quality of untreated water and the permeate are presented. Escherichia coli in the untreated water during the dry season (i.e., June and July) was 2 cfu/100 ml and was <1 cfu/100 ml (undetected) following UF, which complied with the WHO and South African National Standards and Guidelines of <1 cfu/100 ml. During the wet/rainy season (February) total coliform was unacceptably high (>2419.2 cfu/100 ml) before UF. Following UF, it dramatically reduced to acceptable level (7 cfu/100 ml) which is within the WHO recommended level of <10 cfu/100 ml. Additionally, during the wet/rainy season E. coli and enterococci were unacceptably high (40.4 cfu/100 ml and 73.3 cfu/100 ml, respectively) before UF but were completely removed following UF, which are within the WHO and SANS recommended limit. The values for electrical conductivity (EC) and turbidity were constantly within the WHO recommended limits of 300 µS/cm corrected at 25°C and <5 NTU, respectively, before and after UF, during dry season and wet season. This suggests that there is no need for pre-treatment of the water for suspended particles and colloids. Considering these data, it can be concluded that the water is suitable for human consumption, following UF.     

Survival of Mycobacterium avium in Drinking Water Biofilms as Affected by Water Flow Velocity, Availability of Phosphorus, and Temperature

Mycobacterium avium is a potential pathogen occurring in drinking water systems. It is a slowly growing bacterium producing a thick cell wall containing mycolic acids, and it is known to resist chlorine better than many other microbes. Several studies have shown that pathogenic bacteria survive better in biofilms than in water. By using Propella biofilm reactors, we studied how factors generally influencing the growth of biofilms (flow rate, phosphorus concentration, and temperature) influence the survival of M. avium in drinking water biofilms. The growth of biofilms was followed by culture and DAPI (4′,6′-diamidino-2-phenylindole) staining, and concentrations of M. avium were determined by culture and fluorescence in situ hybridization methods. The spiked M. avium survived in biofilms for the 4-week study period without a dramatic decline in concentration. The addition of phosphorus (10 μg/liter) increased the number of heterotrophic bacteria in biofilms but decreased the culturability of M. avium. The reason for this result is probably that phosphorus increased competition with other microbes. An increase in flow velocity had no effect on the survival of M. avium, although it increased the growth of biofilms. A higher temperature (20°C versus 7°C) increased both the number of heterotrophic bacteria and the survival of M. avium in biofilms. In conclusion, the results show that in terms of affecting the survival of slowly growing M. avium in biofilms, temperature is a more important factor than the availability of nutrients like phosphorus.     

Microbiological safety of natural mineral water

Natural mineral water originates from groundwater, an oligotrophic ecosystem where the level of organic matter is low and of a very limited bioavailability. The bacterial populations that evolve in these ecosystems are heterotrophic and in starvation-survival state resulting from an insufficient amount of nutrients; for this reason they enter a viable but non-culturable state. After bottling, the number of viable counts increases rapidly, attaining 10(4)-10(5) colony-forming units ml(-1) within 3-7 days. These bacterial communities, identified by culture or with specific probes, are primarily aerobic, saprophytic, Gram-negative rods. Groundwater sources for natural mineral waters are selected such that they are not vulnerable to fecal contamination. Ecological data, especially the diversity and physiological properties of bacterial communities, are essential together with epidemiological studies in order to perform a risk analysis for natural mineral waters. On a continuing basis, the management of microbial risks has to rely on assessment of the heterotrophic plate count and, more specially, on detection of marker organisms, i.e. the classic fecal contamination indicators that have to be absent, and vulnerability indicators for which the occurrence should be as low as possible. It is also recommended to search regularly, but not routinely, for viral and protozoan pathogens.     

Environmental factors affecting the occurrence of mycobacteria in brook sediments

The occurrence of mycobacteria was studied in aerobic brook sediments from 39 drainage areas in Finland. The culturable counts of mycobacteria were related to climatic conditions, characteristics of the drainage area, chemical characteristics of the sediment and water, culturable counts of other heterotrophic bacteria, and microbial respiration rate in the sediment. The counts of mycobacteria varied from 1·1 × 10² to 1·5 × 10⁴ cfu g⁻¹ dry weight of sediment. They correlated positively with the proportion of the drainage area consisting of peatland, total content of C, content of Pb, microbial respiration rate in the sediment, and chemical oxygen demand of the water. The correlations of the mycobacterial counts with pH of sediment and alkalinity of water were negative. The results of the present sediment study and of the forest soil study published earlier strongly suggest that an increase in acidity increases the counts of mycobacteria and decreases the counts and activity of other heterotrophic bacteria. Mycobacterial counts were more than 100 times higher (per dry weight) in forest soils with pH 3·5–4·3 than in sediments with pH 4·5–6·3.     

Antibiofilm activity of sodium bicarbonate, sodium metaperiodate and SDS combination against dental unit waterline-associated bacteria and yeast

Aim: To determine the effect of sodium bicarbonate (SB), sodium metaperiodate (SMP) and sodium dodecyl sulfate (SDS) combination on biofilm formation and dispersal in dental unit waterline (DUWL)-associated bacteria and yeast. Methods and Results: The in vitro effect of SB, SMP and SDS alone and in combination on biofilm formation and dispersal in Pseudomonas aeruginosa, Klebsiella pneumoniae, Actinomyces naeslundii, and Candida albicans was investigated using a 96-well microtitre plate biofilm assay. The combination showed a broad-spectrum inhibitory effect on growth as well as biofilm formation of both gram-negative and gram-positive bacteria, and yeast. In addition, the SB + SMP + SDS combination was significantly more effective in dispersing biofilm than the individual compounds. The combination dispersed more than 90% of P. aeruginosa biofilm whereas the commercial products, Oxygenal 6, Sterilex Ultra, and PeraSafe showed no biofilm dispersal activity. Conclusion: The composition comprising SB, SMP, and SDS was effective in inhibiting as well as dispersing biofilms in DUWL-associated bacteria and yeast. Significance and Impact of the Study: This study shows that a composition comprising environmentally friendly and biologically safe compounds such as SB, SMP, and SDS has a potential application in reducing DUWL-associated acquired infections in dental clinics.     

Temporal Variations in Heterotrophic Mesophilic Bacteria from a Marine Shallow Hydrothermal Vent off the Island of Vulcano (Eolian Islands, Italy)

Abstract The fluctuations of the total microbial abundance, the culturable heterotrophic bacterial population, and the composition of heterotrophic bacteria were investigated in relation to environmental parameters in a shallow, marine hydrothermal vent off the Island of Vulcano (Eolian Islands, Italy). Standing stock dynamics were studied by measuring the total population of picoplankton by direct count and the population of viable heterotrophic bacteria in water and sediment samples collected monthly. The environmental factors most strongly linked to the total microbial abundance and heterotrophic bacterial populations were pH and H₂S content in water and C/N ratio in sediment samples. The pattern of variation of microbial populations associated with water was different from those associated with sediment. Assessment of the qualitative composition of aerobic heterotrophic bacterial communities was based on 30 morphological and biochemical characteristics for each strain. Numerical analysis was used for an initial survey of the similarity among the isolates. The data were successively used to determine the structure and the metabolic potential of water and sediment bacteria. Metabolic properties varied between water- and sediment-isolated bacteria. Bacteria from water were structurally more diverse, and active in the use of carbohydrates, than those from sediment. Moreover, most of the sediment bacteria were able to grow at a high temperature (60 and 70°C). The fluctuations of bacterial characteristics in relation to environmental parameters present an evident temporal variation in water, but not in the sediment habitat. Received: 13 January 1997; Accepted: 7 August 1997