Survival of protozoa in cooling tower biocides

Protozoa from cooling towers may serve as hosts for legionellae, but such protozoa have not been examined with respect to effects of cooling tower biocides. In this study, two ciliate species,Tetrahymena sp andColpoda sp, and two amoebae species,Vannella miroides andAcanthamoeba hatchetti, were isolated from a cooling tower and tested for survival in the presence of four cooling tower biocides. The protozoa were exposed for 24 h to a thiocarbamate compound, an isothiazolone compound, quaternary ammonium compounds (QAC), and tributyltin neodecanoate with quarternary ammonium compounds (TBT/QAC). After exposure, cells were examined for viability. The highest concentration of each biocide in which cells could survive was compared to the manufacturers' recommended maintenance dosage (MRMD) of the biocides.Tetrahymena andColpoda survived concentrations within the range of the MRMD of thiocarbamate and QAC.Vannella andAcanthamoeba survived concentrations within the MRMD of thiocarbamate, isothiazolone, and QAC.Colpoda cysts andAcanthamoebae cysts remained viable after exposure to concentrations much greater than the MRMD of thiocarbamate, isothiazolone, and QAC. None of the protozoa in any stage could survive the MRMD of TBT/QAC. These results show that protozoa indigenous to cooling towers may survive the recommended concentration of certain biocides, and this information may be important in devising procedures for eradicating hosts for legionellae.     

Thermophillic and thermotolerant fungi isolated from the thermal effluent of nuclear power generating reactors: Dispersal of human opportunistic and veterinary pathogenic fungi

Over a period of a year, samples of water, foam, microbial mat, soil and air were obtained from areas associated with the cooling canal of a nuclear power station. The seventeen sample sites included water in the cooling canal that was thermally enriched and soil and water adjacent to, upstream, downstream and at a distance from the generator. Air samples were taken at the plant and at various disstances from the plant. Fifty-two species of thermotolerant and thermophilic fungi were isolated. Of these, eleven species are grouped as opportunistic Mucorales or opportunistic Aspergillus sp. One veterinary pathogen was also isolated (Dactylaria gallopava):The opportunistic/pathogenic fungi were found primarily in the intake bay, the discharge bay and the cooling canal. Smaller numbers were obtained at both upstream and downstream locations. Soil samples near the cooling canal reflected an enrichment of thermophilous organisms, the previously mentioned opportunistic Mucorales and Aspergillus spp. Their numbers were found to be greater than that usually encountered in a mesophilic environment. However, air and soil samples taken at various distances from the power station indicated no greater abundance of these thermophilous fungi than would be expected from a thermal enriched environment. Our results indicate that there was no significant dissemination of thermophilous fungi from the thermal enriched effluents to the adjacent environment. These findings are consistent with the results of other investigators.     

Feeding by marine fish larvae: developmental and functional responses

Synopsis The relationship between prey consumption rate and prey concentration (functional response), and its change with growth (developmental response) were examined in the laboratory for three species of marine fish larvae: bay anchovy Anchoa mitchilli (Engraulidae), sea bream Archosargus rhomboidalis (Sparidae) and lined sole Achirus lineatus (Soleidae). The major objective was to determine relative predatory abilities of the larvae by fitting feeding rate data to developmental and functional response models. Feeding success, prey capture success, attack rates, handling times and search rates were estimated. Prey consumption rates and attack rates of bay anchovy usually were highest, but at the lowest prey level (50 per liter) first-feeding sea bream larvae had the highest consumption rate. Sea bream could consume prey at near-maximum rates at prey levels lower than those required by the other species. As larvae grew, time searching per attack decreased rapidly for all species, especially at low prey levels. Handling time also decreased, but most rapidly for bay anchovy. Search rates were highest for bay anchovy and lowest for lined sole. Bay anchovy had the best apparent predation ability, but when previous results on larval growth rates, survival rates and growth efficiencies were considered, sea bream larvae were the most efficient predators and the least likely of the three species to be limited by low prey levels.     

Reproductive biology and egg mortality of bay anchovy, <Emphasis Type="Italic">Anchoa mitchilli</Emphasis>, in the Hudson River estuary

Synopsis Bay anchovy, Anchoa mitchilli, is an ecologically important species in coastal systems, acting as predators on zooplankton while serving as prey for many birds and economically important finfish species. Despite the importance of bay anchovy, limited information exists concerning reproductive biology and egg mortality from the northern parts of its range. Spawner demographics suggested Hudson River fish were older and larger than reported elsewhere. Egg mortality rates were higher in the Hudson River than reported in more southerly systems. Differences in the reproductive biology of bay anchovy suggest either anthropogenic factors or latitudinal differences between populations exist and therefore should warrant further investigations on this species in the northern portions of its range     

Occurrence and pathogenicity of Naegleria fowleri in artificially heated waters.

The occurrence of pathogenic Naegleria fowleri in thermal discharges, recipient waters, and cooling towers of eight power plants located in western Pennsylvania was investigated for 2 years in conjunction with several environmental measurements. Pathogenic N. fowleri was detected in one cooling tower and in the discharge, receiving waters, or both of five of eight localities. The occurrence of this organism was related to elevated temperatures, but no significant correlation was found for other biological and chemical parameters. Laboratory experiments on the effect of pH on pathogenic N. fowleri documented 100% survival at a range from 2.1 to 8.15. Higher pH reduced or killed the amoebae. No case of human primary amoebic meningoencephalitis occurred during the study.     

Occurrence and pathogenicity of Naegleria fowleri in artificially heated waters

The occurrence of pathogenic Naegleria fowleri in thermal discharges, recipient waters, and cooling towers of eight power plants located in western Pennsylvania was investigated for 2 years in conjunction with several environmental measurements. Pathogenic N. fowleri was detected in one cooling tower and in the discharge, receiving waters, or both of five of eight localities. The occurrence of this organism was related to elevated temperatures, but no significant correlation was found for other biological and chemical parameters. Laboratory experiments on the effect of pH on pathogenic N. fowleri documented 100% survival at a range from 2.1 to 8.15. Higher pH reduced or killed the amoebae. No case of human primary amoebic meningoencephalitis occurred during the study.     

The effect of temperature and water quality on the in vitro development and survival of Asellus aquaticus (Crustacea: Isopoda) eggs

The rate of development and degree of survival of Asellus aquaticus eggs outside the marsupium of ovigerous females are affected by water quality and temperature. Eggs were maintained in polluted river water and relatively clean canal water. Developmental rates increase with increased temperature, but survival decreases. Eggs from polluted site ovigerous females survive better in clean water than in polluted water. Eggs from clean site ovigerous females maintained in polluted water have significantly lower survival rates than eggs from the polluted site at all temperatures tested. The developmental rate of clean site eggs is increased significantly in polluted water at 10–25 °C, possibly as a response to the stress imposed upon them.It is suggested that the method outlined might form the basis of a useful bioassay technique for measuring water quality.     

Disinfection of bacterial biofilms in pilot-scale cooling tower systems

The impact of continuous chlorination and periodic glutaraldehyde treatment on planktonic and biofilm microbial communities was evaluated in pilot-scale cooling towers operated continuously for 3 months. The system was operated at a flow rate of 10,080 l day^?1. Experiments were performed with a well-defined microbial consortium containing three heterotrophic bacteria: Pseudomonas aeruginosa, Klebsiella pneumoniae and Flavobacterium sp. The persistence of each species was monitored in the recirculating cooling water loop and in biofilms on steel and PVC coupons in the cooling tower basin. The observed bacterial colonization in cooling towers did not follow trends in growth rates observed under batch conditions and, instead, reflected differences in the ability of each organism to remain attached and form biofilms under the high-through flow conditions in cooling towers. Flavobacterium was the dominant organism in the community, while P. aeruginosa and K. pneumoniae did not attach well to either PVC or steel coupons in cooling towers and were not able to persist in biofilms. As a result, the much greater ability of Flavobacterium to adhere to surfaces protected it from disinfection, whereas P. aeruginosa and K. pneumoniae were subject to rapid disinfection in the planktonic state.     

Application of Live Monocells from Macroalgae to Shellfish Seed Production

Monocells were isolated from several macroalgae, Porphyra yezoensis, Undaria pinnatifida, and Laminaria japonica, by digestion with alga-tool enzymes. The monocells were then used to feed the parents or larvae of bay scallop Argopecten irradians, blood cockle Arca inflata, and abalone Haliotis discus juveniles. Results showed that the parents of bay scallop and blood cockle fed with Porphyra monocells could mature and discharge eggs and spermatozoa and their larvae could metamorphose; the survival rate of abalone juveniles fed with isolated cells from Laminaria and Undaria increased by 100% compared with that of those fed with artificial food. Received October 2, 1997; accepted December 18, 1998.     

Tidal transport of the bay anchovy, Anchoa mithchilli, in darkness

Transport of the adult bay anchovy, Anchoa mitchilli , in darkness was studied in laboratory and field experiments. In a hydraulic flume, anchovy schooling ceased in darkness and, at a current speed of 30cms⁻¹, 99% of all fish were transported to the end of the flume. In field experiments, fish marked with neutral red dye and released in a creek at flood tide were recaptured 5–1 km upstream 4 h after release at night, and were recaptured within 200m of the release site 3 h after release in daylight. It is concluded that in the neutral environment, bay anchovy schools break up at night and individual fish are transported by currents, whereas in daylight, bay anchovy schools maintain a relatively fixed location. This nocturnal transport phenomena may also exist in other marine and estuarine pelagic fishes, and may help in understanding behaviour and distribution of peiagic estuarine fishes.     

Survival of Plant Tissue at Super-Low Temperatures. IV. Cell Survival with Rapid Cooling and Rewarming

Thin unmounted cortical tissue sections from winter twigs of the mulberry tree were held with a thin forceps and rapidly immersed in liquid nitrogen from room temperatures without prefreezing. They were rewarmed; rapidly in water at 10° to 40°, or slowly, in air at room temperatures. In those sections rapidly rewarmed, all survived. None survived in those sections rewarmed slowly in air.

Tissue sections mounted between coverglasses with water were extracellulary prefrozen at the temperatures low enough to dehydrate almost all of the freezable water in cells. These sufficiently prefrozen cells could survive immersion in liquid nitrogen, and the survival value was very little affected by the rates of cooling to and rewarming from super-low temperatures. With insufficient prefreezing at higher temperatures, however, the rewarming process seriously influenced the survival value of cells frozen at super-low temperatures. Slow rewarming in air destroyed all of the cells, while rapid rewarming in water at 30° did not affect them. An abrupt decrease in the survival value in insufficiently prefrozen cells during rewarming was also observed at temperatures above approximately −50° following immersion in liquid nitrogen. Very little decrease in the survival value was observed in any of the cells that had been sufficiently prefrozen.

These results indicate that cells which are insufficiently prefrozen may contain freezable water which nucleates during rapid cooling in liquid nitrogen and then grows during the subsequent slow rewarming into ice masses which destroy the viability of the cells. Such fatal intracellular freezing rarely occurs in sufficiently prefrozen cells, irrespective of the rate of cooling to or rewarming from super-low temperatures.

     

Survival of sub-zero temperatures by two South Georgian beetles (Coleoptera,Perimylopidae)

Summary The ability of adults and larvae of two species of perimylopid beetles (Hydromedion sparsutum, Perimylops antarcticus) to survive sub-zero temperatures was studied at Husvik, South Georgia in summer during October–December 1990. Experiments determined their survival at constant sub-zero temperatures, their lower lethal temperatures and individual supercooling points. The effects of cooling rates (0.015°, 0.5° and 2.0°C min^–1) and starvation on survival were also assessed. Mean supercooling points of field-collected individuals of both species were in the range -3.0° to -5.4°C with Perimylops having a deeper capacity (ca. 1.5°C) for supercooling relative to Hydromedion. The former species also survived freezing temperatures significantly better than the latter and its mean lower lethal temperature was 2.5°C lower. At a constant temperature of -8.5°C, the median survival times for Perimylops adults and larvae were 19 and 26 h respectively, whilst both stages of Hydromedion died within 3 h. The three cooling rates resulted in significantly different median survival temperatures for adult Hydromedion with 0.5°C min^–1 producing maximum survival. Prior starvation did not have a significant influence on the survival of either species at sub-zero temperatures although both adults survived less well. The results support field observations on the habitats and distribution of these insects, and suggest differing degrees of freezing tolerance.     

Legionnaires' disease outbreaks and cooling towers with amplifiedLegionella concentrations

This study was designed to investigate the possible association of high colony counts of legionellae from cooling towers and evaporative condensers with Legionnaires' disease outbreaks. We obtained legionellae counts from samples of cooling towers and evaporative condensers that were the likely sources of two different Legionnaires' disease outbreaks and compared these counts with those from cooling towers that were not associated with reports of human disease. Among 675 potential control cooling tower that were samples from 258 facilities, 136 facilities had one or more cooling towers that met our criteria for inclusion into the study. Samples taken from buildings where an outbreak had occurred had much higherLegionella counts than did samples from other buildings. Colony counts from the two outbreak-associated facilities were significantly higher than colony counts from other facilities [Wilcoxon Rank Sum Test (Exact), p<0.01]. The results of the study suggest that, among cooling towers that test positive for the presence of legionellae, higher colony counts are associated with higher risk of Legionnaires' disease.     

The biology of the black goby, Gobius niger L. in an English south-coast bay

A population of Gobius niger in a south-coast bay was sampled over a 12-month period. The bay received the cooling water discharge from Fawley power station. Four year-classes were recorded of which the 1+ group was found to be responsible for the bulk of egg production. The main spawning period was from April to May although some evidence of batch spawning was found. Growth was found to be faster than has been reported for populations elsewhere, possibly because of the raised water temperature. Despite higher temperatures the growing season was not extended beyond that of other comparable black goby populations, being restricted to a 4–5 months period after the main spawning period. Seasonal changes in diet were believed to be the result of the effect of water temperature on fish movement. Feeding occurred throughout the year.     

Rapid cold hardening in the olive fruit fly Bactrocera oleae under laboratory and field conditions

A rapid cold hardening response was studied in females and males of the olive fruit fly Bactrocera (Dacus) oleae. When laboratory-reared females and males were transferred and maintained from the rearing temperature of 24 °C for 2 h to –6.5 °C approximately 5% survived. However, conditioning of both females and males for 2 h at various temperatures from 0 to 10 °C before their exposure for 2 h to –6.5 °C increased survival to 80 to 92%. A similar rapid cold hardening response in both females and males was also induced through gradual cooling of the flies at a rate of approximately 0.4 °C per min. The rapid increase in cold tolerance after prior conditioning of the flies to low temperatures, was rapidly lost when they returned to a higher temperature of 24 °C. In the field, in late February and early March, females and males were capable of a rapid cold hardening response. After exposure to the critical temperature they suffered a high mortality when tested in the afternoon and low mortality early in the morning on consecutive days, probably because of differences in the prevailing field temperatures a few hours before testing. This plasticity of cold tolerance gained through rapid cold hardening may allow the flies to survive during periods of the year with great fluctuation in circadian temperatures.     

Response of <Emphasis Type="Italic">Chrysaora quinquecirrha</Emphasis> medusae to low temperature

Because of their high abundance in Chesapeake Bay, Chrysaora quinquecirrha medusae may be an important reservoir of organic matter. The timing and location of the decomposition of biomass from medusae may have implications for carbon cycling in the bay. Our objective was to identify the cause of C. quinquecirrha medusa disappearance to better understand when and where decomposition occurs. A time series of visual surface counts and vertical net hauls in the Choptank River, a tributary of Chesapeake Bay, showed that as temperatures approached 15°C, C. quinquecirrha medusae disappeared from the surface, but persisted in net hauls until temperatures reached 10°C. In order to test whether medusae sink upon cooling, we exposed C. quinquecirrha medusae to low temperatures in large static tanks and measured their depth and pulsation rates twice daily for at least 6 days. This procedure was repeated three times through the 2008 jellyfish season. On average, individuals exposed to temperatures below 15°C were found deeper and pulsed slower than those in the warmer control tank. This suggests that low temperatures cause the medusae to sink before cooling to the limit of their physiological tolerance and may have implications for the deposition of organic matter associated with the seasonal disappearance of medusae from Chesapeake Bay.     

Adaptation of amoebae to cooling tower biocides

Adaptation of amoebae to four cooling tower Biocides, which included a thiocarbamate compound, tributyltin neodecanoate mixed with quaternary ammonium compounds (TBT/QAC), another QAC alone, and an isothiazolin derivative, was studied. Previously we found that amoebae isolated from waters of cooling towers were more resistant to cooling tower biocides than amoebae from other habitats. Acanthamoeba hatchetti and Cochliopodium bilimbosum, obtained from American Type Culture Collection and used in the previous studies, were tested to determine whether they could adapt to cooling tower Biocides. A. hatchetti was preexposed to subinhibitory concentrations of the four Biocides for 72h, after which they were tested for their resistance to the same and other biocides. C. bilimbosum was exposed to only two biocides, as exposure to the other two was lethal after 72 h. Preexposure to the subinhibitory concentrations of the Biocides increased the resistance of the amoebae, as indicated by a significant increase in the minimum inhibitory concentration (up to 30-fold). In addition, cross-resistance was also observed, i.e., exposure to one biocide caused resistance to other biocides. These results show that amoebae can adapt to biocides in a short time. The phenomenon of cross-resistance indicates that regularly alternating biocides, as is done to control microbial growth in cooling towers, may not be effective in keeping amoeba populations in check. On the contrary, exposure to one biocide may boost the amoebae's resistance to a second biocide before the second biocide is used in the cooling tower. Since amoebae may harbor Legionella, or alone cause human diseases, these results may be important in designing effective strategies for controlling pathogens in cooling towers. Correspondence to: S.G. Berk.     

Effects of biocidal treatments to inhibit the growth of legionellae and other microorganisms in cooling towers.

The effects of biocidal treatments for cooling towers were examined through the use of chemicals and ultraviolet irradiation to inhibit the growth of legionellae and other microorganisms. In the water of cooling towers without continuous biocidal treatments, heterotrophic bacteria and bacterivorous protozoan first appeared, and then legionellae increased up to 10(4) CFU/100 ml. When a UV sterilizer was connected to the cooling tower, the legionellae count was 1/10 or 1/100 of that in the nontreated tower water. In the water of towers supplemented continuously with the biocidal chemicals, legionellae were not found during a 4-month period. The biocidal treatments tested were proved to suppress the increase of legionellae in cooling-tower water, and thus are useful in preventing the outbreak of legionellosis due to inhalation of contaminated aerosol from the cooling tower system.     

Effects of thermal effluents from the Bergum power station on the phytoplankton in the Bergumermeer

Summary The Bergum power station (600 MW) of the Friesian Provincial Electricity Board is situated at the northern shore of the Lake Bergum. The lake has a mean depth of 1.3 m and a surface area of 4.4 km². Its northern half is separated by a break-water into an intake area in the north-west and a discharge area in the north-east.The Lake Bergum is connected with other water bodies in the northern provinces of the Netherlands by four canals. The whole yaer various amounts of water enter Lake Bergum mainly from the western canal (Prinses Margrietkanaal) and to a lesser extent from the southern canal (De Lits). In wet seasons lake water flows off, mainly after passing the power station, to the northern canal (De Zwemmer); then the heated water (22 m³.sec_–1) does not enter the discharge area of the lake. When evapo-transpiration exceeds precipitation lake water flows off mainly to the eastern canal (Kolonelsdiep). In these relatively dry periods most of the heated water returns to the lake in the discharge area.We found that the mean increase in water temperature effected by the condensors of the power station was ca. 5°C; the maximum increase was 7.5°C. On average about 25% of the whole lake had a noticable higher (1°C) temperature than the intake water, only 6.5% was about 2°C above ambient temperatures.For about 3.5 years (1974–Sept. 1978) water samples for analysis of the chlorophyll concentrations of the different areas within the lake and the surrounding canals were taken every week during the growing season, and fortnightly during the winter period. The chlorophyll concentrations of the intake water were about 5% higher than those of the discharge water leaving the power station. Near the mouth of the northern canal in the discharge area still small, but significant lower chlorophyll concentrations were found. The southern half of the lake, in which practically no elevated water temperatures were found, had significant higher chlorophyll concentrations (10–15%) than the intake area. Water entering the lake from the western canal had significant (10–15%) lower chlorophyll concentrations than the intake area of the lake. Probably, relatively chlorophyll-poor canal water and chlorophyll-rich water from the southern lake area mix in the intake area. While the water passes the power station the chlorophyll concentrations decrease. In the discharge area of the lake the chlorophyll concentrations of the discharge water gradually increase again to values equal to those of the intake area.During the last 2 years of the research period oxygen production and consumption experiments were conducted almost every month. In each experiment light and dark botties containing intake and discharge water were suspended in water with both water temperatures. The light intensities during the incubation periods (2–3 hours) were chosen according to maximum production values. The incubations were started within one hour and/or one day after sampling. Directly after sampling gross productivity of the intake water incubated at discharge temperatures was about 1.5 times as high as at intake temperatures. The gross productivity of the discharge water was always somewhat lower than the gross productivity of the intake water incubated at corresponding temperatures. After one day this inhibiting effect of passage through the power station had increased, even when the discharge water had been cooled down to intake temperatures immediately after sampling.The oxygen consumption of the discharge water incubated at discharge temperatures as well as at intake temperatures was about 1.3 times the oxygen consumption of the intake water at intake temperatures. After one day the discharge water, which had stayed at discharge temperatures, consumed 1.6–1.7 times as much as the intake water incubated at intake temperatures. The oxygen consumption of the discharge water which had been cooled down to intake temperatures directly after sampling, was after one day still 1.3 times the oxygen consumption of the intake water at intake temperatures.This research was financially supported by the Ministerie van Volksgezondheld en Millieuhygiëne (Ministry of Public Health and the Environment). An extensive report (in Dutch) will be published this year.     

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.