ALGAL‐RELATED TASTES AND ODORS IN PHOENIX WATER SUPPLY: PRELIMINARY REPORT

Frequent episodes of algal‐related tastes and odors (T & O) in drinking waters in metropolitan Phoenix, Arizona prompted initiation of a three‐year project in July 1999 to investigate the occurrence of T & O metabolites and to develop a comprehensive management strategy to reduce the problems in drinking water supplies in arid environments. Two metabolites, 2‐methylisoborneol (MIB) and geosmin, have been identified as compounds responsible for the earthy‐musty tastes and odors in water supplies. Both were detected in the water supply system, including source rivers, reservoirs, canal delivery system and water treatment plants. Higher concentrations of MIB and geosmin occurred in distribution canals than in the upstream reservoirs indicating that significant production of the T & O compounds occurs within the canal system. A baseline‐monitoring program has been established for the complex water supply system, with special emphasis on the canal system. Efforts are underway to investigate possible correlations between physical/chemical parameters, algal composition and biomass, with the occurrence of MIB and geosmin. In addition, several physical and chemical treatments are planned for the canal system to reduce algal growth and related MIB and geosmin concentrations.     

Causative microorganisms for musty odor occurrence in the eutrophic Lake Kasumigaura

The musty odor phenomenon in Lake Kasumigaura, Japan (a eutrophic lake that is used for various purposes, including water supply source) was examined. The causative microorganisms responsible for the production of the odor compounds, geosmin and/or 2-methylisoborneol (MIB), and the odor release mechanism were studied in vitro and in situ. The numbers of the filamentous P. tenue in the lake water column were closely correlated with the concentration of MIB, but not geosmin. The isolated monoxenic P. tenue in culture produced only MIB. Geosmin concentration was well correlated with the number of actinomycetes in the sediment. Forty isolates of actinomycetes from the sediment around the water supply intake tower produced both geosmin and MIB in culture. Furthermore, the average ratio of production of geosmin to MIB was 1.40–1.0. Actinomycetes in the sediment played an important role in geosmin production. We concluded that geosmin and MIB from actinomycetes in the sediment, and MIB from P. tenue in the water column were jointly responsible for the severe musty odor occurrence of the waters supply source at Lake Kasumigaura.     

Mutagenesis by ethidium bromide and N-methyl-N′-nitro-N-nitrosoguanidine in off-flavour compound producing strains ofStreptomyces

Various species of actinomycetes and cyanobacteria can impart earthy/musty off-flavours to drinking water supplies and to pond-raised fish and other aquatic food animals. The genetic determinants for production of the most common off-flavour compounds [geosmin and 2-methylisoborneol (MIB)] have not been extensively studied. An attempt has been rrlade to study the genetics of production of these compounds was demonstrated by DNA-curing analysis. The effects of two curing agents [ethidium bromide (EB) and N-methyl-N′-nitro-N-nitrosoguanidine (NTG)] on tha loss of linear plasmid DNA and generation of bald mutants (no aerial mycelia) inStreptomyces halstedii andStereptomyces violaceusniger which produce geosmin and MIB, respectively, were observed. Production of earthy/musty odour was not eliminated, but was reduced by 55–95% in the plasmid cured strain. Data suggested that off-flavour production is likely chromosomally-encoded in theseStreptomyces isolates.     

Geosmin production and polyphasic characterization of Oscillatoria limosa Agardh ex Gomont isolated from the open canal of a large drinking water system in Tianjin City,China

Taste and odor (T & O) episodes always cause strong effects on drinking water supply system. Luanhe River diversion into Tianjin City in China is an important drinking water resource. Massive growth of a benthic filamentous cyanobacterium with geosmin production in the open canal caused a strong earthy odor episode in Tianjin. On the basis of the morphological and molecular identification of this cyanobacterium as Oscillatoria limosa Agardh ex Gomont, the genetic basis for geosmin biosynthesis and factors influencing growth and geosmin production of O. limosa CHAB 7000 were studied in this work. A 2268-bp open reading frame, encoding 755 amino acids, was amplified and characterized as the geosmin synthase gene (geo), followed by a cyclic nucleotide-binding protein gene (cnb). Phylogenetic analysis implied that the evolution of the geosmin genes in O. limosa CHAB 7000 might involve a horizontal gene transfer event. Examination on the growth and geosmin production of O. limosa CHAB 7000 at different light intensities showed that the maximum geosmin production was observed at 10 μmol photons m⁻² s⁻¹, while the optimum growth was at 60 μmol photons m⁻² s⁻¹. Under three temperature conditions (15 °C, 25 °C, and 35 °C), the maximum growth and geosmin production were observed at 25 °C. Most amounts of geosmin were retained in cells during the growth phase, but high temperature and low light intensity increased the release of geosmin into the medium, implying that O. limosa CHAB 7000 had a high potential harm for the release of geosmin from its cells at these adverse conditions.     

Differential partitioning of geosmin and 2-methylisoborneol between cellular constituents in Oscillatoria tenuis

A newly isolated strain of Oscillatoria tenuis produces geosmin and 2-methylisoborneol (MIB) simultaneously. The intra- and extracellular distribution of both compounds was studied. Geosmin was much less released to the medium compared to MIB. Different amounts of each of the compounds were in different cellular fractions. The geosmin and MIB in these fractions were recoverable by ammonium sulphate precipitation, suggesting binding to protein. Thylakoid and cytoplasmic membranes contained most geosmin while high MIB protein ratios were found in a colourless protein fraction as obtained by gel exclusion chromatography.Abbreviations MIB 2-methylisoborneol - SDS sodium dedecyl sulphate     

Fate in Model Ecosystems of Microbial Species of Potential Use in Genetic Engineering

Three Oscillatoria strains and one Anabaena species were isolated from three different water supply systems in California that experienced earthy-musty taste and odor problems in their drinking water. Unialgal cultures, free of actinomycetes, were purged using the Grob closed-loop stripping analysis method, and the resulting methylene chloride extracts were analyzed on a gas chromatograph/mass spectrometer. Geosmin was produced by Oscillatoria simplicissima and Anabaena scheremetievi, and 2-methylisoborneol was produced by O. curviceps and O. tenuis. These compounds are the two major causes of earthy-musty tastes and odors in water. In three instances, the major odorant found in culture was previously identified in the water or sediment sample from which the respective organism was isolated. O. curviceps was implicated in a taste and odor episode involving 2-methylisoborneol in a major reservoir. Geosmin and 2-methylisoborneol were easily detected with culture samples of only 4 to 25 ml.     

Numerical models for management of <Emphasis Type="BoldItalic">Anabaena circinalis</Emphasis>

The control of nuisance species of cyanobacteria in reservoirs is a critical issue for the international water industry, as these organisms can produce toxins and compounds that taint potable water with unpleasant tastes and odours. To assist with effective management of cyanobacterial growth, numerical models that are either site specific or universally applicable can be employed. An artificially destratified reservoir was modelled with the coupled hydrodynamic-ecological numerical model DYRESM-CAEDYM. The validation site was Myponga Reservoir, South Australia, a highly managed drinking water supply reservoir. Chemical dosing (CuSO₄) and artificial mixing via an aerator and two raft-mounted mechanical surface mixers (hereafter referred to as surface mixers) are used at Myponga to manage the growth of the scum-forming cyanobacterium Anabaena circinalis. The dominant phytoplankton community was adequately modelled, and combinations of the various management options were simulated whereupon informed operational strategies could be implemented. Without any intervention, permanent stratification would occur and the growth of Anabaena circinalis would peak above 3 μ g L⁻¹, producing geosmin that would exceed the taste and odour threshold (10 ng L⁻¹); with the individual use of the aerator or surface mixers, growth of Anabaena circinalis was significantly reduced to below 1 μ g L⁻¹.     

Relationships between cyanobacterial production and the physical and chemical properties of a Midwestern Reservoir,USA

Drinking water reservoirs in agricultural dominated watersheds are particularly vulnerable to cyanobacterial blooms. A major byproduct of cyanobacteria is the production of objectionable taste and odor compounds such as geosmin. During May 1997 to September 1998, we studied spatial and temporal patterns of cyanobacterial abundance and composition with respect to a series of physical and chemical properties in Clinton Lake, located in east central Kansas, USA. Our results suggest that nutrients (in particular TN, NO₃–N concentrations), turbidity, and hydrologic regime all played potentially important roles in regulating cyanobacterial production. Specifically, low levels of nitrogen coupled with the internal release of phosphorus from the lake sediment under brief periods of anoxia may have helped promote cyanobacterial blooms. There was also a strong association between cyanobacterial blooms, geosmin production, and most taste and odor events in Clinton Lake. Anabaena circinalis appeared to be the source for geosmin production as a result of senescing algal cells just after the primary die-off of cyanobacteria.     

原核生物中土霉味化合物二甲萘烷醇和2-甲基异茨醇生物合成研究进展

由原核生物蓝藻和放线菌引起的水体土霉异味问题备受世界的关注。本文以土霉味化合物二甲萘烷醇和2-甲基异茨醇的研究历史为主线,综述了土霉味化合物二甲萘烷醇和2-甲基异茨醇的化学特性、生物合成途径以及生物合成相关基因和酶等方面的研究进展,探讨了土霉味化合物二甲萘烷醇和2-甲基异茨醇生物合成研究的重要意义,并展望了土霉味化合物二甲萘烷醇和2-甲基异茨醇生物合成研究存在的问题及发展的方向。     

东湖富营养水体中藻菌异味性次生代谢产物的研究

以自制的开环吹脱捕捉（ＯＬＳＡ）装置富集东湖营养水的藻菌异味性次生代谢产物,并对这些化合物进行鉴定和周年测,同时计数水样中的藻、菌生物量。研究发现,东湖水体产生“土霉味”的主要成分为２－甲基异茨醇（ＭＩＢ）,它来源于席藻和放线菌的次生代谢,其随季节变化的规律为一年有５个高峰期,即１月、４月和９月;并且,ＭＩＢ的浓度与水体富营养化程度以及席藻和放线菌的生物量呈正相关。     

Isolation of bacteria capable of growth with 2-methylisoborneol and geosmin as the sole carbon and energy sources

Using a relatively simple enrichment technique, geosmin and 2-methylisoborneol (MIB)-biodegrading bacteria were isolated from a digestion basin in an aquaculture unit. Comparison of 16S rRNA gene sequences affiliated one of the three isolates with the Gram-positive genus Rhodococcus, while the other two isolates were found to be closely related to the Gram-negative family Comamonadaceae (Variovorax and Comamonas). Growth rates and geosmin and MIB removal rates by the isolates were determined under aerated and nonaerated conditions in mineral medium containing either of the two compounds as the sole carbon and energy source. All isolates exhibited their fastest growth under aerobic conditions, with generation times ranging from 3.1 to 5.7 h, compared to generation times of up to 19.1 h in the nonaerated flasks. Incubation of the isolates with additional carbon sources caused a significant increase in their growth rates, while removal rates of geosmin and MIB were significantly lower than those for incubation with only geosmin or MIB. By fluorescence in situ hybridization, members of the genera Rhodococcus and Comamonas were detected in geosmin- and MIB-enriched sludge from the digestion basin.     

Studies on the earthy-musty odours in natural water (IV). Mechanism of earthy-musty odour production of actinomycetes

Many reservoirs for water supply have been troubled with earthy-musty odour compounds--2-metylisoborneol (2-MIB) and geosmin. Both of these compounds are terpenoid and related to the metabolite of L-methionine. An experiment using [CD3]methionine and [14CH3]methionine showed that the C-2 methyl group in 2-MIB originate from L-methionine. In the incubation experiment with 10-1000 mg/l of L-methionine, 2-MIB and geosmin appeared in the earlier stages, in greater amounts than in the control. The maximum production of 2-MIB and geosmin increased considerably in the experiments with 10 and 100 mg/l of L-methionine. The effective time for L-methionine addition was after 1 d. Additions after 3 and 5 d were similar to the control. In the incubation experiment with 10-1000 mg/l of folic acid, 2-MIB and geosmin increased only during the 1000 mg/l addition. There seems to be little doubt that L-methionine takes part in the metabolism of 2-MIB and geosmin.     

Effects of light and temperature on the odor production of 2-methylisoborneol-producing Pseudanabaena sp. and geosmin-producing Anabaena ucrainica (cyanobacteria)

Frequent off-flavor events caused by geosmin and 2-methylisoborneol (MIB) have attracted research on the main producers, cyanobacteria. This study evaluated the effects of light and temperature on the odor production of MIB-producing Pseudanabaena sp. Lauterborn and geosmin-producing Anabaena ucrainica (Schhorb.) Watanabe. The maximum MIB production and lowest growth rate (indicated by the chlorophyll a (Chl a)) were observed at 35 °C compared with that at 10 °C and 25 °C. Cultures grown under a light intensity of 60 μmol photons m⁻² s⁻¹ demonstrated the highest MIB production and minimum growth rate, whereas the minimum MIB production and maximum growth rate were obtained under 10 μmol photons m⁻² s⁻¹. Similar patterns were observed for geosmin production. A. ucrainica had the highest geosmin production and lowest Chl a concentration under 10 °C and 60 μmol photons m⁻² s⁻¹. Moreover, greater proportions of geosmin and MIB were released into extracellular under growth-inhibiting temperatures and light intensities. An inverse correlation between odor production and the cell growth rate was suggested, and this relationship may reflect the competition for substrates of odor and Chl a synthesis. Thus, the accumulation of geosmin and MIB was probably the result of decreased cellular metabolic activity in cyanobacteria.     

The control of diatom and cyanobacterial blooms in reservoirs using barley straw

A potable supply reservoir, with a long history of diatom blooms in spring and cyanobacterial blooms in summer, was treated with barley straw in March 1993 with subsequent additions in December 1993 and June 1994. Within two months of the initial treatment, algal numbers started to fall compared with previous years and have remained consistently lower throughout 1993 and 1994. Cyanobacteria have not bloomed and cell numbers remained low. Chemical analysis of the water showed locally elevated concentrations of geosmin close to the straw on one occasion but the overall concentration of this and a range of other organic molecules remained within acceptable limits and at concentrations similar to those found in other untreated reservoirs in the region. Observed and potential advantages to public health and potable supply management resulting from the use of barley straw are discussed.     

Survival and multiplication of Legionella pneumophila in municipal drinking water systems

Studies were conducted to investigate the survival and multiplication of Legionella spp. in public drinking water supplies. An attempt was made, over a period of several years, to isolate legionellae from a municipal system. Sampling sites included the river water supply, treatment plant, finished water reservoir system, mains, and distribution taps. Despite the use of several isolation techniques, Legionella spp. could not be detected in any of the samples other than those collected from the river. It was hypothesized that this was due to the maintenance of a chlorine residual throughout the system. To investigate the potential for Legionella growth, additional water samples, collected from throughout the system, were dechlorinated, pasteurized, and inoculated with Legionella pneumophila. Subsequent growth indicated that many of these samples, especially those collected from areas affected by an accumulation of algal materials, exhibited a much greater ability to support Legionella multiplication than did river water prior to treatment. Chemical analyses were also performed on these samples. Correlation of chemical data and experimental growth results indicated that the chemical environment significantly affects the ability of the water to support multiplication, with turbidity, organic carbon, and certain metals being of particular importance. These studies indicate that the potential exists for Legionella growth within municipal systems and support the hypothesis that public water supplies may contaminate the plumbing systems of hospitals and other large buildings. The results also suggest that useful methods to control this contamination include adequate treatment plant filtration, maintenance of a chlorine residual throughout the treatment and distribution network, and effective covering of open reservoirs.     

Survival and multiplication of Legionella pneumophila in municipal drinking water systems.

Studies were conducted to investigate the survival and multiplication of Legionella spp. in public drinking water supplies. An attempt was made, over a period of several years, to isolate legionellae from a municipal system. Sampling sites included the river water supply, treatment plant, finished water reservoir system, mains, and distribution taps. Despite the use of several isolation techniques, Legionella spp. could not be detected in any of the samples other than those collected from the river. It was hypothesized that this was due to the maintenance of a chlorine residual throughout the system. To investigate the potential for Legionella growth, additional water samples, collected from throughout the system, were dechlorinated, pasteurized, and inoculated with Legionella pneumophila. Subsequent growth indicated that many of these samples, especially those collected from areas affected by an accumulation of algal materials, exhibited a much greater ability to support Legionella multiplication than did river water prior to treatment. Chemical analyses were also performed on these samples. Correlation of chemical data and experimental growth results indicated that the chemical environment significantly affects the ability of the water to support multiplication, with turbidity, organic carbon, and certain metals being of particular importance. These studies indicate that the potential exists for Legionella growth within municipal systems and support the hypothesis that public water supplies may contaminate the plumbing systems of hospitals and other large buildings. The results also suggest that useful methods to control this contamination include adequate treatment plant filtration, maintenance of a chlorine residual throughout the treatment and distribution network, and effective covering of open reservoirs.     

Studies on the earthy-musty odours in natural water (IV). Mechanism of earthy-musty odour production of actinomycetes

Many reservoirs for water supply have been troubled with earthy-musty odour compounds—2-methylisoborneol (2-MIB) and geosmin. Both of these compounds are terpenoid and related to the metabolite of l-methionine. An experiment using [CD₃]methionine and [¹⁴CH₃]methionine showed that the C-2 methyl group in 2-MIB originate from l-methionine. In the incubation experiment with 10–1000 mg/l of l-methionine, 2-MIB and geosmin appeared in the earlier stages, in greater amounts than in the control. The maximum production of 2-MIB and geosmin increased considerably in the experiments with 10 and 100 mg/l of l-methionine. The effective time for l-methionine addition was after 1 d. Additions after 3 and 5 d were similar to the control. In the incubation experiment with 10–1000 mg/l of folic acid, 2-MIB and geosmin increased only during the 1000 mg/l addition. There seems to be little doubt that l-methionine takes part in the metabolism of 2-MIB and geosmin.     

Reducing harmful algae in raw water by light-shading

The occurrence of harmful algal bloom in water source poses a serious water safety problem to local water supply systems. In order to ensure the raw water quality, the feasibility of an in situ light-shading measure was investigated through enclosure experiment and pilot-scale experiment. The results showed that harmful algal bloom could be controlled by light-shading lasting for 6–9 days, with water quality being partially improved. When aeration was added, the reduction of algal biomass could be enhanced, and water quality was further improved compared to that without aeration. These experimental results offered an attractive in situ algal control measure for lakes or reservoirs suffered from harmful algal bloom.     

Toxic cyanobacteria and drinking water: Impacts,detection, and treatment

Blooms of toxic cyanobacteria in water supply systems are a global issue affecting water supplies on every major continent except Antarctica. The occurrence of toxic cyanobacteria in freshwater is increasing in both frequency and distribution. The protection of water supplies has therefore become increasingly more challenging. To reduce the risk from toxic cyanobacterial blooms in drinking water, a multi-barrier approach is needed, consisting of prevention, source control, treatment optimization, and monitoring. In this paper, current research on some of the critical elements of this multi-barrier approach are reviewed and synthesized, with an emphasis on the effectiveness of water treatment technologies for removing cyanobacteria and related toxic compounds. This paper synthesizes and updates a number of previous review articles on various aspects of this multi-barrier approach in order to provide a holistic resource for researchers, water managers and engineers, as well as water treatment plant operators.