Genotoxicity of microcystic cyanobacteria extract of a water source in China.

The water pollution of toxic cyanobacteria (blue-green algae) is a worldwide problem and worsens with industrialization. Microcystins are potent cyclic heptapeptidic hepatotoxins produced mainly by Microcystis aeruginosa, and their hepatotoxicity has been well-documented. In contrast, information on the genotoxic effects of microcystins is relatively scarce. In our present study, the genotoxicity of microcystic cyanobacteria extract (MCE) of a water source in China was studied using Salmonella typhimurium assay (Ames test), comet assay (Single cell gel electrophoresis) and mouse micronucleus test. Results from Ames test indicated that MCE had strong mutagenicity regardless of the presence of S9. Moreover, MCE was able to induce DNA damage in primary cultured rat hepatocytes examined by comet assay. In addition, MCE also enhanced bone marrow micronucleated polychromatic erythrocytes in mice. The analysis of HPLC showed that the main component of MCE was microcystin-LR. The understanding of the potent genotoxicity of MCE will help to establish the possible link between water cyanobacteria contamination and high risk of primary liver cancer found in some endemic areas.     

Morphological and molecular characterization of cyanobacteria from a Brazilian facultative wastewater stabilization pond and evaluation of microcystin production

The cyanobacterial population in the Cajati waste stabilization pond system (WSP) from São Paulo State, Brazil was assessed by cell isolation and direct microscope counting techniques. Ten strains, belonging to five genera (Synechococcus, Merismopedia, Leptolyngbya, Limnothrix, and Nostoc), were isolated and identified by morphological and molecular analyses. Morphological identification of the isolated strains was congruent with their phylogenetic analyses based on 16S rDNA gene sequences. Six cyanobacterial genera (Synechocystis, Aphanocapsa, Merismopedia, Lyngbya, Phormidium, and Pseudanabaena) were identified by direct microscope inspection. Both techniques were complementary, since, of the six genera identified by direct microscopic inspection, only Merismopedia was isolated, and the four other isolated genera were not detected by direct inspection. Direct microscope counting of preserved cells showed that cyanobacteria were the dominant members (>90%) of the phytoplankton community during both periods evaluated (summer and autumn). ELISA tests specific for hepatotoxic microcystins gave positive results for six strains (Synechococcus CENA108, Merismopedia CENA106, Leptolyngbya CENA103, Leptolyngbya CENA112, Limnothrix CENA109, and Limnothrix CENA110), and for wastewater samples collected from raw influent (3.70 μg microcystins/l) and treated effluent (3.74 μg microcystins/l) in summer. Our findings indicate that toxic cyanobacteria in WSP systems are of concern, since the treated effluent containing cyanotoxins will be discharged into rivers, irrigation channels, estuaries, or reservoirs, and can affect human and animal health.     

Ecologic and hygienic evaluation of Lake Ladoga as a source of drinking water

Eutrophication and pollution of Lake Ladoga cause epidemiologic and toxicologic risks for its use as a supply of drinking water. Increased levels of nutrients (N and P) and low molecular weight organic compounds enhance microbial activity and transformations of xenobiotic compounds in the lake, and promote the formation of chlorinated compounds in drinking water purification process. Experimental studies on the toxicity of water and sediment samples from Priozersk, Pitkärranta, Petrokrepost and Volkhov Bay areas have resulted in marked blood, immune system and genetic responses in laboratory rats. Severe toxicity is also evidenced by Daphnia biotests. Epidemiological studies have revealed elevated morbidity and mortality levels in the human populations in certain regions in the environs of Lake Ladoga, with respect to a group of diseases with a potential connection with water quality, i.e. diseases of digestive organs and genitourinary system as well as malignant neoplasms.     

Divergent responses of diverse microalgae commonly found in drinking water source water to UV-C treatment

Microalgae pose major problems for drinking water treatment processes either through membrane fouling or through blooming events in source water during the summer months. Disinfection treatments such as chlorination and UV-C treatment of drinking water are designed to remove microorganisms including microalgae and bacteria. In order to treat water efficiently it is critical to understand the effect of UV-C treatment on the viability and vitality of phytoplankton. Moreover, this information can be used to assess the utility of UV-C treatment as a potential pre-treatment option for water clean-up. In this study we tested the population growth of 6 phytoplankton species representing three major taxonomic groups (green algae, diatoms and cyanobacteria), commonly found in freshwater lakes in North America and relevant for water treatment facilities. We exposed these species to UV-C treatment and monitored their post-exposure population growth over a 52–55-day period using a plate-based MPN (most probable number) method. We found the green alga Scenedesmus quadricauda and the cyanobacterium Anabaena flos-aquae to be most tolerant to the treatment, while the cyanobacterium Microcystis aeruginosa and the diatom Asterionella formosa were found to be most sensitive with viability being reduced at comparatively lower doses. No significant delays in growth post treatment (i.e. time lags) were reported within species. Asterionella formosa and Aulacoseira granulata showed the longest time lags of all species, which might be in part correlated with DNA repair mechanisms and other physiological adaptations. This work provides the basis for the assessment of UV-C treatment efficacy on freshwater phytoplankton.

     

Microalgae and cyanobacteria as a source of glycosidase inhibitors

Culture filtrates and organic solvent extracts of over 500 freshwater and marine eukaryotic microalgae and cyanobacteria were screened for the presence of glycosidase inhibitors. Rapid colorimetric assays were used to detect inhibitors of alpha-glucosidase, alpha-amylase and beta-galactosidase. Inhibitors were found from 38 species. The results suggest that microalgae and cyanobacteria have potential as a source of glycosidase inhibitors which may have clinical applications.     

Detection of microcystin producing cyanobacteria in Spirulina dietary supplements using multiplex HRM quantitative PCR

Arthrospira species, under the name ‘Spirulina’, are used as food supplement for its protein, vitamins, and minerals which have several health benefits. Cyanobacterial toxins including microcystins can possibly contaminate these dietary supplements causing hepatotoxicity, tumour formation, and other disorders. The safe use of dietary supplements necessitates the need to assess such toxins in the algal food supplement. The methods which evaluate these dietary supplements should be highly sensitive, cost-effective, and rapid. In this study, multiplex HRM qPCR analysis was used to detect microcystin (MC)-producing cyanobacteria in Spirulina dietary supplements. The multiplex HRM qPCR detection limit was found to be 25 ag of mcyB spiked in a standard concentration of pcb (25 pg). Two distinct melt curves characteristic of pcb (Tm 82.8 ± 0.07 °C) and mcyB (Tm 77.9 ± 0.05 °C) were observed. Microcystin contamination was detected only in the fish food supplements and not in human dietary supplements of Spirulina. Liquid chromatography–high-resolution mass spectrometry analysis further confirmed the presence of the congeners of microcystin in the identified positive samples.     

Detection of infectious rotavirus in naturally contaminated source waters for drinking water production

Aims:  To assess chlorine susceptibility of Legionella pneumophila grown from two amoebic hosts, Acanthamoeba castellanii and Hartmannella vermiformis .
Methods and Results:  After being released from amoebae, Leg. pneumophila were chlorinated at 2 and 5 mg l⁻¹ for 5 min–24 h. Bacterial culturability and cytoplasmic membrane deterioration were quantified by culture assay on BCYEα agar and BacLight stains coupled with a fluorescent microscope, respectively. Chlorination reduced the culturability of Leg. pneumophila by 2·93–4·59 log CFU ml⁻¹ and damaged cellular membrane by 53·8–99·2%. Moreover, cells released from H. vermiformis exhibited significantly lower degrees in culturability reduction ( P  = 0·0008) and membrane deterioration ( P  < 0·0001) when compared with those from A. castellanii . The amoebic genus is the most significant parameter affecting cytoplasmic membrane integrity of chlorinated Legionella ( P  < 0·0001), followed by free chlorine concentration ( P  = 0·042).
Conclusions:  Legionella pneumophila replicated from H. vermiformis possess greater chlorine resistance than the cells from A. castellanii .
Significance and Impact of the Study:  This study shows the heterogeneity of amoebae-grown Leg. pneumophila in chlorine susceptibility, which should be considered in the control of legionellae proliferation, particularly in the systems where H. vermiformis is dominant, e.g. hot water plumbing.     

Isolation and characterization of microcystin producing Microcystis from a Central Indian water bloom

Microcystis aeruginosa Kütz, a well-known microcystin (hepatotoxin) producing cyanobacterium was the dominant bloom-forming organism in a mesotrophic lake at Nagpur in Central India, which was isolated and characterized for morphospecies and microcystin content. Compact spherical colonies, formation of daughter colonies, and clathration of older colonies leading to release of solitary cells, were characteristics of laboratory grown M. aeruginosa. Its growth, monitored as increase in optical density (OD) measured at 678 nm (the wavelength selected using dilution curve technique), exhibited a maximum specific growth rate (μ_max) of 0.34 day⁻¹ which, was attained on the 5th day of the experiment with a doubling time of 3.25 days. Though the morphological characters of the M. aeruginosa under field conditions were not retained under laboratory conditions, the microcystin content and type of variants did match with bloom samples. Reverse phase high performance liquid chromatography (RP-HPLC) analyses revealed that the laboratory grown isolate of Microcystis produced microcystin-RR (732 μg g⁻¹ dry weight biomass) and demethylated microcystin-RR (165 μg g⁻¹ dry weight biomass) variants, which are reported to be less toxic when compared to microcystin-LR. LC/ESI/MS further confirmed the presence of these two variants. Geographical distribution of microcystin variants and their prevailing concentrations need to be considered during formulation of guideline values for drinking and recreational waters.     

Investigation of microcystin removal from eutrophic surface water by aquatic vegetable bed

In southern China, many freshwater ecosystems, including lakes, rivers and reservoirs, are eutrophic. The nutrient loading coupled with year-round warm weather favors the growth of cyanobacteria, several of which can produce cyanotoxins, especially the potent liver toxins called microcystins, which are often detected in eutrophic drinking water sources. For purifying raw water used as source of drinking water treatment plants, an aquatic vegetable bed (AVB) experiment had been carried out in a hypertrophic waterfront of Lake Taihu, China, since October 2002. AVB was a simplification of the nutrient film technique (NFT) used to produce vegetables, which requires large quantities of water and nutrients. The average removal rates of total microcystin-RR and microcystin-LR were 63.0% and 66.7%, respectively. This study indicated that Ipomoea aquatica was able to absorb microcystins by using enzyme-linked immunosorbent assay (ELISA), and that the roots absorbed more toxins than leaves and stems. We used fluorescence in situ hybridization (FISH) to analyze the density of microcystin-degrading bacteria in AVB sediment. Two species of microcystin-degrading bacteria were detected, which indicated that microcystin bio-degradation processes did occur in AVB. Protozoa and metazoa were abundant in the rhizosphere. Aspidisca sp., Vorticella sp., Philodina sp., and Lecane sp. were the dominant species. The predation function of protozoa and metazoa had a positive effect on removal of cyanobacteria and microcystins.     

Nitrate removal from drinking water in a fluidized-bed biological denitrification bioreactor

In most industrially developed countries an increasing degree of nitrification can be observed in potential water reservoirs. High nitrate content is unacceptable by public health standards. Since contamination seems to be unavoidable, the only realistic solution is purification prior to utilization. One of the potential variations is biological denitrification with a highly intensive facility, a fluidized- or expanded-bed bioreactor. Based on laboratory and pilot plant experiments, a detailed analysis is presented on the problems arising and solutions offered in the construction of a purification system meeting high quality requirements of drinking water purification. The crucial point is selection of the denitrifying microorganisms and organic matter required for denitrification, which simultaneously determines the attachment of bacteria to the support material (autoimmobilization), the intensity of nitrate removal and the character of post-purification.     

Cyanobacteria and cyanotoxins in the source water from Lake Chivero,Harare, Zimbabwe,and the presence of cyanotoxins in drinking water

The phytoplankton community and cyanotoxins in Lake Chivero (formerly Lake McIlwaine) and the presence of cyanotoxins in treated drinking water were investigated between 2003 and 2004. A typical seasonal succession of Cyanobacteria species occurred from January to April, Bacillariophyta from May to July, and Cryptophyta and Chlorophyta from August to December. Microcystis aeruginosa and M. wesenbergii, known producers of the toxin microcystin, and the non-toxic cyanobacterium M. novacekii dominated during summer. The highest concentrations of microcystins and lipopolysaccharide endotoxins occurred when cyanobacterial biomass was highest. Lipopolysaccharide endotoxin concentrations in the lake ranged between 8 and 3 200 Endotoxin Units (EU) ml^?1. Microcystin concentrations in treated water were below the recommended safe limit for drinking water. Lipopolysaccharide endotoxin concentrations in treated water ranged from 0.15 to 11 EU ml^?1. The phytoplankton community comprised non-microcystin-producing species for the greater part of the study period.     

Biosorptive removal of arsenic from drinking water

A biomass derived from the plant Momordica charantia has been found to be very efficient in arsenic(III) adsorption. An attempt was made to use this biomass for arsenic(III) removal under different conditions. The parameters optimized were contact time (5-150 min), pH (2-11), concentration of adsorbent (1-50 g/l), concentration of adsorbate (0.1-100mg/l), etc. It was observed that the pH had a strong effect on biosorption capacity. The optimum pH obtained for arsenic adsorption was 9. The influence of common ions such as Ca(2+), Mg(2+), Cd(2+), Se(4+), Cl(-), SO(4)(2-), and HCO(3)(-), at concentrations varying from 5 to 1000 mg/l was investigated. To establish the most appropriate correlation for the equilibrium curves, isotherm studies were performed for As(III) ion using Freundlich and Langmuir adsorption isotherms. The pattern of adsorption fitted well with both models. The biomass of M. charantia was found to be effective for the removal of As(III) with 88% sorption efficiency at a concentration of 0.5mg/l of As(III) solution, and thus uptake capacity is 0.88 mg As(III)/gm of biomass. It appears that this biomass should be used as a palliative food item. Further it also appears that the dietary habits may play a role in the toxic effects of ingested arsenic.     

70 Use of molecular probe to detect taste/odor-causing cyanobacteria in the phoenix drinking water distribution system

Upper Klamath Lake (UKL) is the largest lake in Oregon (area 287 km², avg. depth 4.2 m). It is naturally eutrophic and regularly suffers nuisance summer blooms of cyanobacteria, principally Aphanizomenon flos-aquae (AFA). Sediment coring studies show that AFA was absent or minimal until about 1880 when a steady increase began, culminating in the blooms of recent decades. These studies show concomitant increases in sediment N (∼20%) and P (∼50%) along with shifts in the algal flora indicating increased eutrophication. These changes correlate with increased human impacts, such as deforestation, construction, roadbuilding etc., and especially the ditching, diking and draining of adjacent wetlands for conversion to agriculture. Agricultural nutrient runoff, especially P, has been often cited as the cause of the AFA blooms, and most attention has been focused on the dynamics of UKL during the summer bloom. We propose that a more significant factor may be the loss of early-season suppression of AFA because of the loss of the lake-associated wetlands, which originally constituted 42% of the lake area, and which have declined in area by 66.3% since the late 1800's. The melting of snow and ice in the spring would flush into the lake a surge of wetland plant decomposition products, most significantly organic acids and humic substances. We propose that formerly these wetland effluents caused a complex of effects on lake pH, solar UV transparency, photochemical interactions, nutrient availability, and Daphnia grazing dynamics, which would have combined to prevent the development of any AFA bloom.     

Nitrate removal from drinking water using a membrane-fixed biofilm reactor

Biological treatment of drinking water is a cost-effective alternative to conventional physico/chemical processes. A new concept was tested to overcome the main disadvantage of biological denitrification, the intensive post-treatment process to remove microorganisms and remnant carbon source. The biological reaction zone and carbon supply were separated from the raw water stream by a nitrate-permeable membrane. Denitrification takes place in a biofilm, which is immobilized at the membrane. In a series of bench-scale runs, different types of membranes and reactor configurations were investigated. The best denitrification rates achieved were 1230 mg NO₃ ⁻-N m⁻² day⁻¹. In one run, raw water containing 100 mg NO₃ ⁻ l⁻¹ was completely freed from nitrate. The membrane and the attached biofilm also represent a barrier against the passage of the C source and nutrients into the raw water. At concentrations of 20 mg l⁻¹ ethanol and 15 mg l⁻¹ phosphate in the bioreactor no diffusion through the membrane into the treated water was observed. Without any post-treatment, the effluent met nearly all the relevant criteria for drinking water; only the colony count was slightly increased. Received: 18 December 1996 / Received last revision: 14 April 1997 / Accepted: 19 April 1997     

Tangential-flow ultrafiltration with integrated inhibition detection for recovery of surrogates and human pathogens from large-volume source water and finished drinking water

Tangential-flow ultrafiltration was optimized for the recovery of Escherichia coli, Enterococcus faecalis, Clostridium perfringens spores, bacteriophages MS2 and PRD1, murine norovirus, and poliovirus seeded into 100-liter surface water (SW) and drinking water (DW) samples. SW and DW collected from two drinking water treatment plants were then evaluated for human enteric viruses.     

Microbiological quality of drinking water from dispensers in Italy

Background  

Water coolers are popular in office buildings and commercial stores and the quality of this source of drinking water has the potential to cause waterborne outbreaks, especially in sensitive and immunocompromised subjects. The aim of this study was to determine the quality of water plumbed in coolers from commercial stores in comparison with tap water in Italy.