The effect of ozone onLegionella pneumophila and other bacterial populations in cooling towers

Ozone was found to reduce the numbers of bacteria detectable by plate counts and the numbers of presumptiveLegionella pneumophila (DFA-reactive cells), including those that were INT positive, in test cooling towers. The numbers of DFA-reactiveL. pneumophila eventually reached those of the makeup water (Troy, NY, city water). Microbial slime on the interior tower surfaces andPseudomonas populations in the tower waters were also reduced. Water chemistry parameters measured showed no tendency toward a condition that would cause accelerated deterioration of tower material.     

Legionella spp. in Puerto Rico cooling towers

Water samples from air conditioning cooling towers receiving different treatment protocols on five large municipal buildings in San Juan, P.R., were assayed for various Legionella spp. and serogroups by using direct immunofluorescence. Several water quality parameters were also measured for each sample. Guinea pigs were inoculated with water samples to confirm pathogenicity and recover viable organisms. Legionella pneumophila serogroups 1 to 6, L. bozemanii, L. micdadei, L. dumoffii, and L. gormanii were observed in at least one of the cooling towers. L. pneumophila was the most abundant species; its density reached 10(5) cells per ml, which is within the range that is considered potentially pathogenic to humans. A significantly higher density of L. pneumophila was observed in the cooling tower water that was not being treated with biocides. Percent respiration (INT) and total cell activity (acridine orange direct count) were inversely correlated with bacterial density. This study demonstrates that Legionella spp. are present in tropical air-conditioning cooling systems and that, without continuous biocide treatment, they may reach densities that present a health risk.     

Legionella spp. in Puerto Rico cooling towers.

Water samples from air conditioning cooling towers receiving different treatment protocols on five large municipal buildings in San Juan, P.R., were assayed for various Legionella spp. and serogroups by using direct immunofluorescence. Several water quality parameters were also measured for each sample. Guinea pigs were inoculated with water samples to confirm pathogenicity and recover viable organisms. Legionella pneumophila serogroups 1 to 6, L. bozemanii, L. micdadei, L. dumoffii, and L. gormanii were observed in at least one of the cooling towers. L. pneumophila was the most abundant species; its density reached 10(5) cells per ml, which is within the range that is considered potentially pathogenic to humans. A significantly higher density of L. pneumophila was observed in the cooling tower water that was not being treated with biocides. Percent respiration (INT) and total cell activity (acridine orange direct count) were inversely correlated with bacterial density. This study demonstrates that Legionella spp. are present in tropical air-conditioning cooling systems and that, without continuous biocide treatment, they may reach densities that present a health risk.     

Distribution of Legionella longbeachae serogroup 1 and other legionellae in potting soils in Australia.

Legionella longbeachae serogroup 1 and other Legionella spp. were isolated from 73% of 45 potting soils made in Australia by 13 manufacturers but were not detected in 19 potting soils made in Greece, Switzerland, and the United Kingdom examined between March 1989 and May 1990. Several Legionella species were isolated from a small number of samples of uncomposted pine sawdusts, but it is not known whether sawdust was the source of some of the legionellae found in potting soils. Legionella spp. persisted for periods ranging from 3 to 10 months in a potting soil held at temperatures between -20 and 35 degrees C. Isolates of L. longbeachae serogroup 1 from soil did not grow at 43 degrees C, a temperature which was also lethal for this species in soil. Most Legionella spp. isolated from potting and natural soils belonged to one distinct group according to analysis of ubiquinones and were serologically related to several known species in this group. A small number of potting soils contained L. pneumophila and L. micdadei.     

An alkaline approach to treating cooling towers for control of Legionella pneumophila.

Earlier field and laboratory studies have shown that Legionella species survive and multiply in the pH range 5.5 to 9.2. Additionally, the technical feasibility of operating cooling towers at elevated alkalinities and pH has previously been documented by published guidelines. The guidelines indicate that these conditions facilitate corrosion control and favor chlorine persistence which enhances the effectiveness of continuous chlorination in biofouling control. This information suggests that control of Legionella species in cooling towers can be accomplished by operating the towers under alkaline conditions. To test this possibility, we collected water samples over a period of months from a hospital cooling tower. The samples were analyzed for a variety of chemical parameters. Subsamples were pasteurized and inoculated with non-agar-passaged Legionella pneumophila which had been maintained in tap water. Correlation of subsequent Legionella growth with corresponding pH and alkalinity values revealed statistically significant inverse associations. These data support the hypothesis that operating cooling towers outside of the optimal conditions for Legionella growth (e.g., at elevated alkalinities and a pH greater than 9) may be a useful approach to controlling growth in this habitat.     

Altered rate of synthesis of specific peptides in the legionellae in response to growth temperature

The effect of a sharp temperature rise (30° to 42°C) on the rate of peptide synthesis inLegionella pneumophila (three strains),Tatlockia micdadei (Tatlock), andFluoribacter bozemanae (WIGA) was examined. Most striking was an increase in the rate of synthesis of 4–5 peptides of approximate molecular weight 85,000 daltons (85k), 78k, 70k, and either 60k (only in theL. pneumophila strains and WIGA) or 62.5k, and 40k (only in Tatlock). The response required new mRNA synthesis. In addition, the heat shock response of these organisms continued as long as the high temperature condition persisted (at least 24 h) and for at least 1 h after temperature downshift (to 30°C).     

Failure of Legionella pneumophila sensitivities to predict culture results from disinfectant-treated air-conditioning cooling towers.

The disinfection of cooling towers based on manufacturers' treatment protocols, as employed in units installed at various public gathering places in Dallas, Tex. (hotels, municipal auditorium), and at the city health department, was evaluated for effectiveness in controlling Legionella pneumophila and compared with previous laboratory studies. In specimens collected in September and December, 1978, L. pneumophila was isolated from 2 of 4 specimens from untreated cooling towers, 2 of 4 specimens from towers treated with agents deemed ineffective in earlier laboratory tests, 6 of 11 specimens from towers treated with putatively effective agents, and 0 of 4 specimens from towers treated with an agent unknown efficacy. These results suggest the need for further studies to identify biocidal agents effective in eliminating L. pneumophila from air-conditioning cooling towers.     

Legionella in industrial cooling towers: monitoring and control strategies

Aims: Legionella contamination of industrial cooling towers has been identified as the cause of sporadic cases and outbreaks of legionellosis among people living nearby. To evaluate and control Legionella contamination in industrial cooling tower water, microbiological monitoring was carried out to determine the effectiveness of the following different disinfection treatments: (i) continuous chlorine concentration of 0·01 ppm and monthly chlorine shock dosing (5 ppm) on a single cooling tower; (ii) continuous chlorine concentration of 0·4 ppm and monthly shock of biocide P3 FERROCID 8580 (BKG Water Solution) on seven towers. Methods and Results: Legionella spp. and total bacterial count (TBC) were determined 3 days before and after each shock dose. Both strategies demonstrated that when chlorine was maintained at low levels, the Legionella count grew to levels above 10⁴ CFU l^?1 while TBC still remained above 10⁸ CFU l^?1. Chlorine shock dosing was able to eliminate bacterial contamination, but only for 10–15 days. Biocide shock dosing was also insufficient to control the problem when the disinfectant concentration was administered at only one point in the plant and at the concentration of 30 ppm. On the other hand, when at a biocide concentration of 30 or 50 ppm was distributed throughout a number of points, depending on the plant hydrodynamics, Legionella counts decreased significantly and often remained below the warning limit. Moreover, the contamination of water entering the plant and the presence of sediment were also important factors for Legionella growth. Conclusions: For effective decontamination of outdoor industrial cooling towers, disinfectants should be distributed in a targeted way, taking into account the possible sources of contamination. Significance and Impact of the Study: The data of the research permitted to modify the procedure of disinfection for better reduce the water and aerosol contamination and consequently the exposure risk.     

Distribution of Legionella longbeachae serogroup 1 and other legionellae in potting soils in Australia

Legionella longbeachae serogroup 1 and other Legionella spp. were isolated from 73% of 45 potting soils made in Australia by 13 manufacturers but were not detected in 19 potting soils made in Greece, Switzerland, and the United Kingdom examined between March 1989 and May 1990. Several Legionella species were isolated from a small number of samples of uncomposted pine sawdusts, but it is not known whether sawdust was the source of some of the legionellae found in potting soils. Legionella spp. persisted for periods ranging from 3 to 10 months in a potting soil held at temperatures between -20 and 35 degrees C. Isolates of L. longbeachae serogroup 1 from soil did not grow at 43 degrees C, a temperature which was also lethal for this species in soil. Most Legionella spp. isolated from potting and natural soils belonged to one distinct group according to analysis of ubiquinones and were serologically related to several known species in this group. A small number of potting soils contained L. pneumophila and L. micdadei.     

Integrin-like proteins in Candida spp. and other microorganisms

The vertebrate integrins provide a paradigm for cell surface proteins involved in adhesion and morphogenesis. However, homologs of integrins have been found in more primitive organisms. This review will discuss the evidence for surface proteins in Candida albicans and Candida tropicalis that contain motifs reminiscent of integrins and will analyze the contributions of one of these proteins, Int1p, to adhesion, morphogenesis, and virulence. Other microorganisms thought to express integrin-like proteins will also be addressed.     

An alkaline approach to treating cooling towers for control of Legionella pneumophila

Earlier field and laboratory studies have shown that Legionella species survive and multiply in the pH range 5.5 to 9.2. Additionally, the technical feasibility of operating cooling towers at elevated alkalinities and pH has previously been documented by published guidelines. The guidelines indicate that these conditions facilitate corrosion control and favor chlorine persistence which enhances the effectiveness of continuous chlorination in biofouling control. This information suggests that control of Legionella species in cooling towers can be accomplished by operating the towers under alkaline conditions. To test this possibility, we collected water samples over a period of months from a hospital cooling tower. The samples were analyzed for a variety of chemical parameters. Subsamples were pasteurized and inoculated with non-agar-passaged Legionella pneumophila which had been maintained in tap water. Correlation of subsequent Legionella growth with corresponding pH and alkalinity values revealed statistically significant inverse associations. These data support the hypothesis that operating cooling towers outside of the optimal conditions for Legionella growth (e.g., at elevated alkalinities and a pH greater than 9) may be a useful approach to controlling growth in this habitat.     

Effect of phosphate availability on biofilm formation in cooling towers

Abstract

Phosphate limitation has been suggested as a preventive method against biofilms. P-limited feed water was studied as a preventive strategy against biofouling in cooling towers (CTs). Three pilot-scale open recirculating CTs were operated in parallel for five weeks. RO permeate was fed to the CTs (1) without supplementation (reference), (2) with supplementation by biodegradable carbon (P-limited) and (3) with supplementation of all nutrients (non-P-limited). The P-limited water contained ≤10?µg PO₄ l^?1. Investigating the CT-basins and coupons showed that P-limited water (1) did not prevent biofilm formation and (2) resulted in a higher volume of organic matter per unit of active biomass compared with the other CTs. Exposure to external conditions and cycle of concentration were likely factors that allowed a P concentration sufficient to cause extensive biofouling despite being the limiting compound. In conclusion, phosphate limitation in cooling water is not a suitable strategy for CT biofouling control.     

Comparison of polymerase chain reaction and conventional culture for the detection of legionellae in cooling tower waters in Singapore

A total of 80 cooling tower water samples were investigated for legionellae using both cultural and polymerase chain reaction (PCR) methods. PCR was performed with the Perkin Elmer EnviroAmp^™ Legionella kit. Forty-seven samples (58·8%) were found positive by both methods ; 29 samples (36·3%) were positive by PCR only, while four samples (5%) showed PCR inhibition despite the adoption of the more stringent sample preparation protocol especially designed to eliminate inhibitors.     

Phototrophic biofilms on the interior walls of concrete Iterson-type cooling towers

Algae and cyanobacteria form noticeable biofilms in the open spaces of natural draught cooling towers. In spite of this, our knowledge of these communities is still poor. During this study, phototrophic biofilms of seven cooling towers at four different plants in the Czech Republic were collected and studied in order to obtain information on the composition of the communities of these biofilms. Fresh material and cultures of collected material were analyzed with light microscopy. Twenty-five cyanobacterial and four algal taxa were found.     

Relative sensitivities of environmental legionellae to selective isolation procedures.

A survey of water samples to determine the efficacy of standard procedures for the isolation of environmental legionellae was conducted. Marked variations in intraspecies resistance to selective agents and treatments were observed, and in experiments with one of the isolates, the response was modified by culture conditions. Five selective procedures incorporating acid (pH 2.2) and heat (50 degrees C, 30 min) treatments, with and without plating on buffered charcoal-yeast extract agar supplemented with vancomycin (5 micrograms/ml), polymyxin B (60 U/ml), and cycloheximide (80 micrograms/ml), caused 5 to 99% decreases in viable counts of pure cultures in water suspensions. The differences in the responses of the cultures to the five treatments were statistically significant. Cells in retained samples of naturally contaminated water from which the original cultures had been isolated were significantly less sensitive than artificially grown isolates. The sensitivities of the laboratory-grown cells to the treatments were affected by the length of incubation on buffered charcoal-yeast extract agar. Whereas acid resistance increased after 24 h of incubation, resistance to the antibiotic mixture decreased.     

Field trial of biocides for control of Legionella in cooling towers

Legionella and amebae populations in 16 cooling towers were challenged with three commercially available biocide formulations. The active agents were: a chlorinated phenolic thioether (CPTE), bromo-nitro-propane-diol (BNPD), and bromo-chloro-dimethylhydantoin (BCD, in briquette form). The towers were dosed with these biocides for approximately 4 weeks. BCD was effective against Legionella in each of nine challenge experiments, and CPTE in eight of nine challenges. BNPD was effective in only five of 11 challenges. None of the biocides had any significant effect in reducing planktonic amebae concentrations during the challenges.     

Development of a new seminested PCR method for detection of Legionella species and its application to surveillance of legionellae in hospital cooling tower water.

The presence of PCR inhibitors in water samples is well known and contributes to the fact that a practical PCR assay has not been developed for legionella surveillance. In this study, we devised a new seminested PCR assay for detection of Legionella spp. in water samples as a means of overriding the PCR inhibitors without loss of sensitivity. The seminested PCR assay utilized primers to amplify the 16S rRNA gene (LEG primers) of 39 Legionella spp. The assay was specific to legionellae, and the sensitivity was 1 fg of extracted Legionella DNA in laboratory examination. To evaluate the feasibility and sensitivity of the PCR assay in identifying the presence of legionellae, it was used to survey Legionella contamination in the water of 49 cooling towers of 32 hospitals. A commercially available EnviroAmp Legionella kit and a culture method were also used in the survey for comparison with the seminested PCR assay. The detection rates of legionellae in the samples were 91.8% (45 of 49) by the PCR assay and 79.5% (39 of 49) by the culture method. The EnviroAmp kit revealed that 30.6% of the water samples (15 of 49) contained inhibitors of the PCR amplification. However, the seminested PCR assay could produce the Legionella-specific DNA bands in 14 of the 15 samples. Although 8 of the 14 samples were positive in the first-step PCR, 6 of the 14 samples became positive in the second-step PCR. These results suggest that the effect of PCR inhibitors in samples, if any, can be reduced because of the dilution of the sample in the second-step PCR and that sensitivity of detection can be increased by the second-step PCR. Thus, the seminested PCR assay with LEG primers to amplify the 16S rRNA gene of 39 Legionella spp. was a practical and sensitive method to detect Legionella spp. in water samples.     

A quantitative method for determining the efficacy of algicides in industrial cooling towers

Summary Quantitative sampling of periphyton from natural substrates is difficult and uncommon due to the nonhomogenous and irregular nature of most natural substrates. This paper describes an experimentally verified method for quantitative sampling of periphyton directly from the relatively homogenous and regular upper deck of a cooling tower.