A role for calcium hydroxide and dolomite in water: acceleration of the reaction under ultraviolet light.

Organic environmental pollutants are now being detected with remarkably high frequency in the aquatic environment. Photodegradation by ultraviolet light is sometimes used as a method for removing organic chemicals from water; however, this method is relatively inefficient because of the low degradation rates involved, and more efficient methods are under development. Here we show that the removal of various organic pollutants can be assisted by calcined dolomite in aqueous solution under irradiation with ultraviolet light. It was possible to achieve substantial removal of bisphenol A, chlorophenols, alkylphenols, 1-naphthol and 17ß-estradiol. The major component of dolomite responsible for the removal was calcium hydroxide. Our results demonstrate that the use of calcium hydroxide with ultraviolet light irradiation can be a very effective method of rapidly removing organic environmental pollutants from water. This is a new role for calcium hydroxide and dolomite in water treatment.

Establishment of evaluation method to determine effects of veterinary medicinal products on manure fermentation using small-scale composting apparatus.

To evaluate on a laboratory scale the influence of veterinary medicinal products (VMPs) excreted into feces on manure fermentation, we have developed an evaluation method that uses a small-scale composting apparatus. Each run is of approximately 3 kg scale and the operation can be conducted in an environmentally controlled laboratory. The main evaluation parameter is calorific value generated by aerobic fermentation. At the sulfadimethoxine (SDM) trial, the volume of CO(2) generated during fermentation and the disappearance of the inhibitory effect of immature manure on sprouting (using Komatsuna (Brassica rapa var. perviridis)) were measured. In addition, DNA of 16S rRNA was extracted from a manure sample and subjected to denaturing gradient gel electrophoresis (DGGE). The results suggest that the presence of such VMPs in feces affected the microbial community in manure fermentation, and indicate that the evaluation method may be used as a standard method to evaluate the effect of VMPs on the microbial community. Using the method, we obtained data of the influence of five VMPs approved for stockbreeding in Japan on swine manure fermentation. Erythromycin (EM) affected the calorific value even at a relatively low concentration (105 mg/3 kg manure). In contrast, oxytetracycline hydrochloride (OTC), norfloxacin (NFLX), and tylosin tartrate (TS) had no effect at that concentration. These VMPs also affected the increase of fermentation temperature when added at high concentrations.

A novel method using cyanobacteria for ecotoxicity test of veterinary antimicrobial agents.

The effect of antimicrobial agents for veterinary use on the growth of cyanobacteria was investigated by measuring minimum inhibitory concentration, medium effective concentration (EC50), and no-observed-effect concentration of seven antimicrobial agents for eight cyanobacteria. The results demonstrated that the seven antimicrobial agents, even at low concentrations, inhibited the growth of cyanobacteria. Microcystis aeruginosa and Synechococcus sp. had the highest sensitivity to the antimicrobial agents used in the present study. It is considered that the utilization of cyanobacteria would enable easy and highly sensitive assessment of the toxicity of such chemicals as antimicrobial agents. We suggest that cyanobacteria be used for ecotoxicity test in addition to the hitherto established method that uses green algae.

Rapid quantification of bacterial cells in potable water using a simplified microfluidic device.

A simplified microfluidic device for quantification of bacteria in potable water was fabricated and examined. Comparisons of counts of Escherichia coli by the microfluidic system and by epifluorescence microscopy closely correlated (r2=0.99). Bacteria in natural mineral water and in purified household tap water were accurately enumerated by using this system within 15 min after fluorescent staining.

Improvement of selective removal of heavy metals in cyanobacteria by NaOH treatment.

In the freshwater cyanobacterium, Tolypothrix tenuis, treatment with 0.1 M NaOH increased its Cd-selective adsorption ability in the presence of Ca(2+) or Mg(2+). The selective adsorption was also achieved by other alkaline treatments. Energy-distributed spectroscopy analysis revealed that Cd(2+) was found mainly on the surface of non-treated cells, whereas it was distributed throughout the cell after NaOH treatment. The alkaline treatment was effective in increasing the selective adsorption ability of the cyanobacterium for other bivalent heavy metals such as Cu(2+), Pb(2+) and Zn(2+). The treatment was also applicable to Anabaena variabilis and Microcystis aeruginosa, which are typical cyanobacteria causing algal blooms. The main binding site of Cd(2+) in NaOH-treated cells is assumed to be the carboxyl groups because the binding ability of the cells was diminished by the esterification of carboxyl groups. These results suggest that alkaline treatment of cyanobacteria is a useful technique for producing biosorbents having highly specific binding abilities for heavy metals.

Biodegradation of bisphenol A and disappearance of its estrogenic activity by the green alga Chlorella fusca var. vacuolata.

Bisphenol A (BPA) is known as an endocrine disruptor and often is found in landfill leachates. Removal of BPA by green alga, Chlorella fusca, was characterized, because we previously found that various phenols were well removed by this strain, including BPA. Chlorella fusca was able to remove almost all BPA in the concentration range from 10 to 80 microM for 168 h under continuous illumination at 18 W/m2. At the low light intensity of 2 W/m2, 82% of 40 microM BPA was removed, and only 27% was removed in the dark. Moreover, C. fusca could remove 90% of 40 microM BPA under the 8:16-h light:dark condition, which was almost as high as that under the continuous-light condition. The amount of BPA contained in the cells was less than the amount of BPA removed from the medium. Monohydroxybisphenol A was detected as an intermediate of BPA degradation. Moreover, estrogenic activity that originated from BPA in the culture medium also completely disappeared. Based on these results, BPA was finally degraded to compounds having nonestrogenic activity. Therefore, C. fusca can be considered a useful organism to remove BPA from landfill leachates.

Evaluation of antimicrobial agents for veterinary use in the ecotoxicity test using microalgae.

The influence of antimicrobial agents approved as veterinary drugs in Japan on the growth of green algae, Selenastrum capricornutum and Chlorella vulgaris, was studied in accordance with the OECD guidelines for testing chemicals. Among the agents tested, growth inhibitory activity was very varied, i.e. erythromycin showed the strongest activity (EC50, 50% effective concentration, = 0.037 mg/l), sulfa drugs had activity to some extent (EC50s of sulfamethoxazole, sulfadiazine, and sulfadimethoxine were 1.5, 2.2, and 2.3 mg/l, respectively), but ampicillin and cefazolin did not inhibit growth (EC50s>1000 mg/l). We also investigated synergistic effect of combining sulfa drugs with trimethoprim or pyrimethamine, which are commonly used as a combined drug. By adding trimethoprim, the growth inhibitory activity of sulfamethoxazole and sulfadiazine was significantly enhanced. Growth inhibition by sulfa drugs was reduced by the addition of folic acid, indicating that they inhibit folate synthesis in green algae.

Degradation of carbendazim and 2,4-dichlorophenoxyacetic acid by immobilized consortium on loofa sponge.

A fungicide, carbendazim (methyl-2-benzimidazole carbamate; MBC), and a herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D), could be simultaneously degraded by a microbial consortium obtained from several soil samples in Japanese paddy fields with enrichment continuous culture. The degradation ability of the consortium was increased by immobilization on loofa (Luffa cylindrica) sponge in comparison with that of free-living consortium. MBC and 2,4-D were completely degraded within 5.5 d and 1.5 d, respectively. The toxicity of these pesticides in culture medium to Daphnia magna was reduced by treatment with the consortium corresponding to their degradation. The degradation ability of the immobilized consortium at pHs in the range from 6 to 9, at temperatures from 15 degrees C to 37 degrees C, and at low NH(4)(+)-N concentrations (1-10 mg/l) was not very different from that under the basal condition (pH 7, 30 degrees C, 424 mg/l NH(4)(+)-N and 472 mg/l PO(4)(3)(-)-P). At low pHs 4 and 5, the ability was significantly lower than that of the basal condition. Moreover, incubation at low PO(4)(3)(-)-P concentrations (1-10 mg/l) caused a decrease in pH at which the degradation ability also became lower. However, the ability in this culture completely recovered when pH was adjusted to 7 or the phosphate concentration was increased to the basal level. Analysis by denaturing gradient gel electrophoresis (DGGE) showed the whole population of the consortium became faint at low pH or low phosphate concentrations but became distinct again as much as those under the basal conditions, indicating that the decrease in the degradation ability caused by low pH was due to that whole population of the consortium underwent serious damage but could survive and recover. These results suggest the immobilized consortium on loofa sponge is a promising material for bioremediation of polluted water with these pesticides in paddy fields.

Photosynthesis-dependent removal of 2,4-dichlorophenol by Chlorella fusca var. vacuolata.

Of 7 green algae, Chlorella fusca var. vacuolata removed about 23% of 2,4-dichlorophenol (DCP) at 10-80 microM after 4 d when grown photoautotrophically. Removal of DCP was growth-dependent and was suppressed dose-dependently by the photosynthesis inhibitor, 3-(3,4-dichlorophenyl)-1,1-dimethyl urea.

Removal of hazardous phenols by microalgae under photoautotrophic conditions.

Various algae were screened for their ability to decrease the concentration of 2,4-dinitrophenol (DNP), as a model compound of hazardous phenols, under photoautotrophic conditions. Chlorella fusca var. vacuolata and Anabaena variabilis grew well and showed high DNP removal ability over the concentration range of 5 to 40 microM. Their abilities to remove various phenols were investigated. More than 90% of 40 microM o- and m-nitrophenol and DNP was removed during the cultivation period of 5 d. o-, p-Chlorophenol and 2,4-dichlorophenol could be removed, but not to a significant extent. C. fusca also removed 85% of bisphenol A, suspected to be an endocrine disrupter. It was found that microalgae would be applicable to the removal of hazardous phenols without the addition of any organic carbon sources.

Effect of algal extract on H2 production by a photosynthetic bacterium Rhodobium marinum A-501: analysis of stimulating effect using a kinetic model.

We have established a system for hydrogen (H2) production from algal starch via lactic acid using a mixed culture of a lactic acid bacterium, Lactobacillus amylovorus, and a photosynthetic bacterium, Rhodobium marinum A-501. We found that the H2 production from lactate was stimulated in the presence of algal extract, which was obtained from algal biomass homogenate used as a substrate in the system by removing settleable solids including starch. To analyze the stimulating effect of algal extract on H2 production, we developed a kinetic model for H2 production by R. marinum A-501. The model revealed that approximately 20% of lactate was consumed for cell mass production, and the remaining portion was a source of reducing power to drive hydrogen production or other cellular processes. In the presence of algal extract, the model indicated that the conversion efficiency from lactate to the reducing power increased from 0.56 to 0.80 and nitrogenase activity increased up to twofold, resulting in the increase in yield of hydrogen from lactate from 29% to 48%. These results suggest that algal extract can attenuate the limitation process in lactate catabolism by which the supplementation of reducing power to drive H2 production was suppressed.