Isolation of a sulfide-producing bacterial consortium from cooling-tower water: Evaluation of corrosive effects on galvanized steel.

Sulfidogenic Clostridia and sulfate reducing bacteria (SRB) often cohabit in nature. The presence of these microorganisms can cause microbially influenced corrosion (MIC) of materials in different ways. To investigate this aspect, bacteria were isolated from cooling tower water and used in corrosion tests of galvanized steel. The identity of the isolates was determined by comparative sequence analysis of PCR-amplified 16S rDNA gene fragments, separated by denaturing gradient gel electrophoresis (DGGE). This analysis showed that, in spite of the isolation process, colonies were not pure and consisted of a mixture of bacteria affiliated with Desulfosporosinus meridiei and Clostridium sp. To evaluate the corrosive effect, galvanized steel coupons were incubated with a mixed culture for 4, 8, 24, 72, 96, 168, 360 and 744 h, along with a control set in sterile culture medium only. The corrosion rate was determined by weight loss, and biofilm formation and corroded surfaces were observed by scanning electron microscopy (SEM). Although the sulfide-producing bacterial consortium led to a slight increase in the corrosion of galvanized steel coupons, when compared to the previous studies it can be said that Clostridium sp. can reduce the corrosive effect of the Desulfosporosinus sp. strain.

Microbial contamination of dental unit waterlines and effect on quality of indoor air.

The microbiological quality in dental unit waterlines (DUWLs) is considered to be important because patients and dental staff with suppressed immune systems are regularly exposed to water and aerosols generated from dental units (DUs). Opportunistic pathogens like Pseudomonas, Legionella, Candida, and Aspergillus can be present in DUWLs, while during consultations, bioaerosols can be dispersed in the air, thus resulting in effects on microbiological quality of indoor air. This present study represents microbiological air and water quality in dental offices (DOs) and also concerns the relationship between the quality of DO air and dental unit water. This study aimed to assess both the microbial quality of dental unit water and the indoor air in 20 DOs and to survey the effect on the quality of the indoor air with the existing microorganisms in dental unit water. Fourteen out of 20 (70 %) DUWLs were found to be contaminated with a high number of aerobic mesophilic heterotrophic bacteria. In terms of bacterial air contamination levels, in 90 % of DOs, a medium level (<500 colony-forming units (CFU)/m(3)) of contamination was determined, while in terms of microfungal air contamination, in all DOs, a low level (<100 CFU/m(3)) of contamination was determined. Potential infection or allergen agents, such as Pseudomonas, Micrococcus, Staphylococcus, Alternaria, Cladosporium, Penicillium, Aspergillus, and Paecilomyces were isolated from water and air samples. This study's determination of contamination sources and evaluation of microbial load in DOs could contribute to the development of quality control methods in the future.

The corrosion behaviour of galvanized steel in cooling tower water containing a biocide and a corrosion inhibitor.

The corrosion behaviour of galvanized steel in cooling tower water containing a biocide and a corrosion inhibitor was investigated over a 10-month period in a hotel. Planktonic and sessile numbers of sulphate reducing bacteria (SRB) and heterotrophic bacteria were monitored. The corrosion rate was determined by the weight loss method. The corrosion products were analyzed by energy dispersive X-ray spectroscopy and X-ray diffraction. A mineralized, heterogeneous biofilm was observed on the coupons. Although a biocide and a corrosion inhibitor were regularly added to the cooling water, the results showed that microorganisms, such as SRB in the mixed species biofilm, caused corrosion of galvanized steel. It was observed that Zn layers on the test coupons were completely depleted after 3 months. The Fe concentrations in the biofilm showed significant correlations with the weight loss and carbohydrate concentration (respectively, p < 0.01 and p < 0.01).

Pioneer colonizer microorganisms in biofilm formation on galvanized steel in a simulated recirculating cooling-water system.

Some bacteria have a higher tendency to produce biofilm than others. Especially, Pseudomonas and Aeromonas strains are acknowledged to be pioneer colonizers and are predominant in biofilm formation. We examined biofilm formation and first attachment maintance of biofilms of Pseudomonas spp., Pseudomonas aeruginosa, Aeromonas spp, sulphate reducing bacteria and filamentous fungi. A simulated recirculating cooling-water system was used. Heterotrophic bacteria counts on galvanized steel and glass surfaces rose during the tidy period of 720 hours. In addition, we determined that although Pseudomonas spp., Pseudomonas aeruginosa and Aeromonas spp. were the pioneer colonizers, they surprisingly could not be determined in the biofilms on both types of surface after 456 hours. Sulphate reducing bacteria were observed in biofilms on both surfaces from the outset of the experiments. Filamentous fungi were seen on the galvanized steel and glass surfaces after 0.5 h.

Comparison of the microbial load of incoming and distal outlet waters from dental unit water systems in Istanbul.

This is a cross-sectional study of the incoming and distal outlet water quality from 41 dental units in Istanbul, carried out to compare the total microbial loads using traditional culture method versus epifluorescence microscopy. The possible presence of Legionella pneumophila using traditional culture method was also analyzed. One hundred and twenty three samples were taken from the high-speed handpiece lines, air-water syringe lines and source (incoming) water supplies from 41 dental units. The samples were assayed for live/dead bacteria, heterotrophic bacterial counts and presence of L. pneumophila bacteria. Thirty nine out of 41 dental units (91%) were not able to meet the standard limit of 200 CFU/ml in dental unit waters. The live bacterial counts were 1-1.5 orders of magnitude higher than aerobic mesophilic heterotrophic bacteria. L. pneumophila (serogroup 2-14) was isolated from five out of 41 units. Some dental units were using commercially bottled (19 l) drinking water as a source. The source water of eight dental unit was heavily contaminated which were fed up by commercially bottled drinking water.

Profiling of environmental Legionella pneumophila strains by randomly amplified polymorphic DNA method isolated from geographically nearby buildings.

Legionella pneumophila (L. pneumophila) which is also known as etiologic agent Legionnaires Disease lives in natural water and man made water systems. These bacteria belonging to Legionellaceae family are divided 15 serogroups. Phenotypical methods used for the identification of Legionella isolates are not very discriminatory. In this study we investigated genotypic features of eight L. pneumophila serogroup 1 and 18 L. pneumophila serogroup 2-14 strains isolated from different buildings in Istanbul by randomly amplified polymorphic DNA (RAPD) method. Eight L. pneumophila serogroup 1 strains (37.5%) were similar RAPD profile and they were isolated from buildings located in a short distance (about 500 m). Four L. pneumophila serogroup 2-14 strains (22%) were identical genotypically. Three of these strains were isolated from buildings located in a short distance.

Microbial contamination of dental unit waterlines in Istanbul, Turkey.

The water used in dental unit waterlines (DUWLs) acts as a coolant for the high-speed equipment and as an irrigant during dental treatments. There are kind of water tanks. DUWLs provide a favorable environment for microbial biofilm and multiplation primarily due to the high surface in the tubing and the character of fluid dynamics in narrow, smooth-walled waterlines. Biofilms can harbour opportunist pathogens such as Legionella sp., Pseudomonas sp. Several studies have shown that DUWLs have high levels of microbial contamination. Presence of high level of microbial contamination is an important problem for dentists and dental patients who are immunocompromised. We collected water samples from DUWLs of 20 private dental offices. We have determined that only 2 (3.4%) out of 59 dental unit water samples were found to meet the standard (<200 CFU.ml(-1)) for DUWLs water quality by American Dental Association (ADA). Of the 59 water samples examined, 14 (24%) were positive for Pseudomonas sp. and 18 (30.5%) were positive for fungi. The most common 14 bacterial strains and seven fungi were isolated. Of bacterial strains, 57.1% were identified: Majority of the bacterial species isolated from our samples was identified as Pseudomonas fluorescens, Pasteurella haemolytica, Photobacterium damsela, Ochrobacter anthropi, Moraxella sp., Aspergillus flavus, Penicillium expansum. Legionella sp. were not detected in all water samples.

Monitoring of biofilm-associated Legionella pneumophila on different substrata in model cooling tower system.

Cooling towers have the potential to develop infectious concentrations of Legionella pneumophila. Legionella counts increases where biofilm and warm water temperatures are present. In this study, biofilm associated L. pneumophila and heterotrophic bacteria were compared in terms of material dependence. Model cooling tower system was experimentally infected by L. pneumophila standard strain and monthly monitored. Different materials were tested for a period of 180 days. The lowest L. pneumophila and heterotrophic plate counts were measured on plastic polymers, whereas L. pneumophila and heterotrophic bacteria were accumulated rapidly on galvanized steel surfaces. It can be concluded that selection of plastic polymers, as a manufacturing material, are suitable for recirculating water systems.

Isolation and identification of enterococci from seawater samples: assessment of their resistance to antibiotics and heavy metals.

A hundred Enterococcus strains were isolated from seawater samples collected from coastal areas of Istanbul. Isolates were identified to the species level using standard biochemical tests specified by Facklam and Collins. The species distribution was as follows Enterococcus faecalis (96%), Enterococcus gallinarum (3%) and Enterococcus solitarius (1%). The resistance of bacteria to both heavy metals (zinc [Zn], iron [Fe], cadmium [Cd], chrome [Cr], cobalt [Co]) and antibiotics (ampicillin 10 microg [AP], penicillin G 10 Units [PG], gentamycin 10 microg [GM], streptomycin 10 microg [S], chloramphenicol 10 microg [C], erythromycin 15 microg [E], kanamycin 30 microg [K], amikacin 30 microg [AK], nalidixic acid 30 microg [NA], and vancomycin 30 microg [VA]) was evaluated. None of the strains was resistant to VA. It was found that among the 100 isolates, those that exhibit resistance to antibiotics, particularly NA, S and K, were also resistant all the heavy metals tested. To our knowledge this is the first report focusing on determination of resistance of environmental enterococci found in Istanbul against heavy metals and antibiotics. Thus, combined expressions of antibiotic and heavy metal resistance may help to reinforce ecological and epidemiological studies and to determine the role of these strains in antibiotic and heavy metal resistance dissemination.

Studies on the efficacy of Chloramine T trihydrate (N-chloro-p-toluene sulfonamide) against planktonic and sessile populations of different Legionella pneumophila strains.

Effectiveness of Chloramine T trihydrate (N-chloro-p-toluene sulfonamide) on both planktonic and sessile populations of different Legionella pneumophila strains was assessed. Although Chloramine T is a recommended commercial formulation for disinfecting cooling towers, there is a lack of published data about the efficacy of this compound against both planktonic and sessile populations of L. pneumophila. Planktonic L. pneumophila strains were suspended in tap water and sessile L. pneumophila strains were grown on stainless steel which is used in the construction of cooling towers, followed by exposure to the biocide. The sensitivity of both planktonic and sessile populations of L. pneumophila strains was different. The biocide was found effective below recommended dosages (0.1-0.3%) against planktonic populations of L. pneumophila, whereas it was determined that higher dosages than those recommended were required for sessile populations of L. pneumophila. The results indicated that studying only the planktonic populations of L. pneumophila for biocide tests might not be sufficient to provide information about the optimum dosage and contact time. Therefore, efficacy has to be tested on both planktonic and sessile bacteria.

Characterization of sulfate reducing bacteria isolated from cooling towers.

In this study, the occurrence and metabolic capacities of sulfate reducing bacteria (SRB) were studied in 36 water samples taken from cooling towers of 30 different buildings, such as hotels and business centres in Istanbul. SRB were present in 14 cooling towers out of 30 (46.6%) buildings and while the lowest concentration of SRB was 10 cells/mL, the highest concentration was determined as 10(4) cells/mL. After the distribution of SRB within cooling towers was determined, several strains of SRB were isolated and characterized metabolically. The isolated strains were composed of vibroid cells, growing anaerobically by using sulfate as electron acceptor and lactate or pyruvate as electron donor. They could be related to the genus Desulfovibrio. In addition, the recorded temperature of water samples was between 12 and 33 degrees C and a significant relationship between the number of SRB and the water temperatures was not found.

Enumeration of Legionella pneumophila in cooling tower water systems.

Legionella pneumophila, the causative agent of Legionnaires' disease, is known to colonise and frequently grow in cooling tower waters. Disease is acquired by inhaling aerosol contaminated by legionellae. Determination of the count of Legionella pneumophila in cooling tower waters may, therefore, be useful for risk assessment. In our survey, 103 water samples from 50 cooling towers were examined over a five-year period to indicate the seasonal distribution and the ecology of L. pneumophila, as regards temperature and pH. L. pneumophila serogroup 1 was found in 44% of the isolated strains, which is primarily responsible for the majority of Legionnaires' disease. The large majority of examined towers had levels of L. pneumophila in the high-risk category. These cooling towers have been linked to many outbreaks of Legionnaires' disease.