Capsid-Damaging Effects of UV Irradiation as Measured by Quantitative PCR Coupled with Ethidium Monoazide Treatment.

The damage to a viral capsid after low-pressure (LP) and medium-pressure (MP) UV irradiation was assessed, using the quantitative or quantitative reverse transcription PCR coupled with ethidium monoazide treatment (EMA-PCR). After UV irradiation, adenovirus 5 (Ad5) and poliovirus 1 (PV1) were subjected to a plaque assay, PCR, and EMA-PCR to investigate the effect of UV irradiation on viral infectivity, genome damage, and capsid damage, respectively. The effectiveness of UV wavelengths in a viral genome and capsid damage of both PV1 and Ad5 was also further investigated using a band-pass filter. It was found that an MPUV lamp was more effective than an LPUV lamp in inactivating Ad5, whereas there was no difference in the case of PV1. The results of viral reduction determined by PCR and EMA-PCR indicated that MP UV irradiation damaged Ad5 capsid. The damage to PV1 and Ad5 capsid was also not observed after LP UV irradiation. The investigation of effects of UV wavelengths suggested that UV wavelengths at 230-245 nm have greater effects on adenovirus capsid in addition to viral genome than UV wavelengths beyond 245 nm.

Characterization of monooxygenase gene diversity in benzene-amended soils.

AIM: To understand soil benzene monooxygenase gene diversity by clone library construction and microarray profiling. METHODS AND RESULTS: A primer set was designed, and benzene monooxygenase gene diversity was characterized in two benzene-amended soils. The dominant sequence types in the clone libraries were distinct between the two soils, and both sequences were assigned to novel clusters. Monooxygenase gene richness and diversity increased after benzene degradation. Oligonucleotide probes for microarray analysis were designed to detect a number of sequenced clones and reported monooxygenase genes. The microarray detected several genes that were not detected in the clone libraries of the same samples. Six probes were detected in more than one soil. CONCLUSIONS: The primer set designed in this study successfully detected diverse benzene monooxygenase genes. The level of diversity may have increased because the degradation of benzene differed from soil to soil. Microarrays have great potential in the comprehensive detection of gene richness as well as the elucidation of key genes for degradation. SIGNIFICANCE AND IMPACT OF THE STUDY: This study introduces a new primer set that may be used to identify diverse benzene monooxygenase genes in the environment; moreover, it demonstrates the potential of microarray technology in the profiling of environmental samples.

Acetate uptake efficiency of polyphosphate-accumulating organisms under exposure to surfactants.

We investigated the influence of surfactants such as linear alkylbenzene sulfonates (LAS) and alcohol ethoxylates (AE) on acetate uptake by polyphosphate-accumulating organisms (PAOs) under anaerobic conditions, using the phosphate requirement for acetate uptake (+DeltaP/-DeltaAc ratio). In order to estimate the +DeltaP/-DeltaAc ratio, anaerobic batch tests were conducted using activated sludge collected from an anaerobic/oxic sequencing batch reactor used to treat municipal wastewater continuously supplemented with a detergent containing LAS and AE. We demonstrated that LAS and AE have both positive and negative impacts on acetate uptake by PAOs. The disadvantage is that long-term exposure to the detergent inhibits acetate uptake by PAOs, thus deteriorating the efficiency, even if the surfactants are no longer present during the tests. Furthermore, the existence of LAS and/or AE with acetate further diminishes the efficiency. The advantage is that LAS and AE are potential sources of polyhydroxyalkanoate for PAOs, because acetate is produced from the surfactants under anaerobic conditions.

Effects of rainfall on the occurrence of human adenoviruses, total coliforms, and Escherichia coli in seawater.

A two-month survey was conducted in order to evaluate the effects of rainfall on the fate of microorganisms in seawater in the Tokyo Bay, Japan. The seawater sample (1,000 mL) was applied to a method to concentrate virus, followed by a quantification of human adenoviruses using the real-time PCR. Total coliforms and E. coli, which were determined by the colony forming method, were detected in all 47 seawater samples, while human adenoviruses were detected in 38 (81%) of the samples. The concentration of tested microorganisms showed 1-2 log units increase after rainfall events, followed by the gradual decrease to the level before the rainfall within a few days.

Direct and indirect inactivation of Microcystis aeruginosa by UV-radiation.

Excessive algal growth in drinking water sources like lakes and reservoirs is responsible for filter-clogging, undesirable taste and odor, disinfection-by-product formation and toxin generation. Although various methods are currently being used to control algal bloom, their successes are limited. Some water utilities routinely use copper sulfate to control excessive algal growth. But there is a growing concern against its use mainly because it is non-specific to target algae and kills many non-target species. In this study, the scope of using UV-radiation to control algal growth was assessed using Microcystis aeruginosa as test species. A UV-dose of 75 mW s cm(-2) was found to be lethal to M. aeruginosa. A smaller dose of 37 mW s cm(-2) prevented growth for about 7 days. It was found that UV-radiation may increase the specific gravity of the cells and thus may adversely affect the ability of the cells to remain in suspension. Three days after a UV-dose of 75 mW s cm(-2), almost all the cells settled to the bottom of the incubation tubes, whereas all the unirradiated cells remained in suspension. It was also observed that UV-radiation on algal extracellular products has a significant residual effect and can contribute to algal growth control. The extent of residual effect depends on the UV-dose and can continue even for 7 days. UV-radiation was found to produce H2O2 in the microM level concentration. But at such level, H2O2 itself is not likely to cause the residual effect that was found in this study.