Use of Preservative Agents and Antibiotics for Increased Poliovirus Survival on Positively Charged Filters

Environmental surveillance of poliovirus (PV) and other non-enveloped viruses can help identify silent circulation and is necessary to certify eradication. The bag-mediated filtration system is an efficient method to filter large volumes of environmental waters at field sites for monitoring the presence of viruses. As filters may require long transit times to off-site laboratories for processing, viral inactivation or overgrowth of bacteria and fungi can interfere with virus detection and quantification (Miki and Jacquet in Aquatic Microb Ecol 51(2):195–208, 2008). To evaluate virus survival over time on ViroCap^? filters, the filters were seeded with PV type 1 (PV1) and/or MS2 and then dosed with preservatives or antibiotics prior to storage and elution. These filters were stored at various temperatures and time periods, and then eluted for PV1 and MS2 recovery quantification. Filters dosed with the preservative combination of 2% sodium benzoate and 0.2% calcium propionate had increased virus survival over time when stored at 25 °C, compared to samples stored at 25 °C with no preservatives. While elution within 24 h of filtration is recommended, if storage or shipping is required then this preservative mixture can help preserve sample integrity. Addition of an antibiotic cocktail containing cephapirin, gentamicin, and Proclin^? 300 increased recovery after storage at 4 and 25 °C, when compared to storage with no antibiotics. The antibiotic cocktail can aid sample preservation if access to appropriate antibiotics storage is available and sample cold chain is unreliable. This study demonstrated that the use of preservatives or antibiotics is a simple, cost-effective method to improve virus detection from ViroCap cartridge filters over time.     

DNA Heat Treatment for Improving qPCR Analysis of Human Adenovirus in Wastewater

PCR inhibitory substances in complex sample matrices can cause false negatives or under-estimation of target concentration. This study assessed DNA heat treatment for reducing inhibition during qPCR analysis of human adenovirus (HAdV) in wastewater samples. Inhibition was reduced by heat treating DNA, where mean HAdV concentration was increased by 0.71 log₁₀ GC/L (and up to 3.04 log₁₀ GC/L in one case), and replicate variability and false negatives were reduced. DNA heat treatment should be further investigated for improving reliability of HAdV concentration estimates in water, which can support more accurate assessment of health risks associated with viral pathogen exposure.     

Assessment and Evaluation of an Integrated Hybrid Anaerobic–Aerobic Sewage Treatment System for the Removal of Enteric Viruses

The capability of a cost-effective and a small size decentralized pilot wastewater treatment plant (WWTP) to remove enteric viruses such as rotavirus, norovirus genogroup I (GGI), norovirus genogroup II (GGII), Hepatitis E virus (HEV), and adenovirus was studied. This pilot plant is an integrated hybrid anaerobic/aerobic setup which consisted of anaerobic sludge blanket (UASB), biological aerated filter (BAF), and inclined plate settler (IPS). Both the UASB and BAF are packed with a non-woven polyester fabric (NWPF). Results indicated that the overall log₁₀ reductions of enteric viruses’ genome copies through the whole system were 3.1 ± 1, 3.3 ± 0.5, and 2.6 ± 0.9 log₁₀ for rotavirus, norovirus GGI, and adenovirus, respectively. Reduction efficiency for both norovirus GGII and HEV after the different treatment steps could not be calculated because there were no significant numbers of positive samples for both viruses. The overall reduction of rotavirus infectious units through the whole system was 2.2 ± 0.8 log₁₀ reduction which is very close to the overall log₁₀ reduction of adenovirus infectious units through the whole system which was 2.1 ± 0.8 log₁₀ reduction. There was no considerable difference in the removal efficiency for different rotavirus G and P types. Adenovirus 41 was the only type detected in the all positive samples. Although the pilot WWTP investigated is cost effective, has a small footprint, does not need a long distance network pipes, and easy to operate, its efficiency to remove enteric viruses is comparable with the conventional centralized WWTPs.     

Interface-mediated synthesis of monodisperse ZnS nanoparticles with sulfate-reducing bacterium culture

We have created a new method of ZnS nanospheres synthesis. By interface-mediated precipitation method (IMPM), monodisperse ZnS nanoparticles was synthesized on the particle surface of sulfate-reducing bacterium nutritious agar culture. Sulfate-reducing bacterium (SRB) was used as a sulfide producer because of its dissimilatory sulfate reduction capability, meanwhile produced a variety of amino acids acting as templates for nanomaterials synthesis. Then zinc acetate was dispersed into nutritious agar plate. Subsequently agar plate was broken into particles bearing much external surface, which successfully mediated the synthesis of monodisperse ZnS nanoparticles. The morphology of monodisperse ZnS nanospheres and SRB were examined by scanning electron microscopy (SEM), and the microstructure was investigated by X-ray diffraction (XRD). The thermostability of ZnS nanoparticles was determined by thermo gravimetric-differential thermo gravimetric (TG-DTG). The maximum absorption wavelengh was analysed with an ultraviolet-visible spectrophotometer within a range of 199–700 nm. As a result, monodisperse ZnS nanoparticles were successfully synthesized, with an average diameter of 80 nm. Maximum absorption wavelengh was 228 nm, and heat decomposed temperature of monodisperse ZnS nanoparticles was 596°C.     

Interface-mediated synthesis of monodisperse ZnS nanoparticles with sulfate-reducing bacterium culture

We have created a new method of ZnS nanospheres synthesis. By interface-mediated precipitation method (IMPM), monodisperse ZnS nanoparticles was synthesized on the particle surface of sulfate-reducing bacterium nutritious agar culture. Sulfate-reducing bacterium (SRB) was used as a sulfide producer because of its dissimilatory sulfate reduction capability, meanwhile produced a variety of amino acids acting as templates for nanomaterials synthesis. Then zinc acetate was dispersed into nutritious agar plate. Subsequently agar plate was broken into particles bearing much external surface, which successfully mediated the synthesis of monodisperse ZnS nanoparticles. The morphology of monodisperse ZnS nanospheres and SRB were examined by scanning electron microscopy (SEM), and the microstructure was investigated by X-ray diffraction (XRD). The thermostability of ZnS nanoparticles was determined by thermo gravimetric-differential thermo gravimetric (TG-DTG). The maximum absorption wavelengh was analysed with an ultravioletvisible spectrophotometer within a range of 199-700 nm. As a result, monodisperse ZnS nanoparticles were successfully synthesized, with an average diameter of 80 nm. Maximum absorption wavelengh was 228 nm, and heat decomposed temperature of monodisperse ZnS nanoparticles was 596℃.     

发挥城市基层组织作用,有效进行公众参与工作

根据作者从事环境评价工作的经验,结合北京市的城市建设项目的案例,说明在公众参与工作中结合中国城市基层组织特点,发挥基层组织作用,能够有效使公众参与环境保护、体现公众环境知情权、制止群体事件的发生,进一步做好公众参与工作。     

Detection Method for Avian Influenza Viruses in Water

Recent events have shown that humans may become infected with some pathogenic avian influenza A viruses (AIV). Since soil and water, including lakes, rivers, and seashores, may be contaminated by AIV excreted by birds, effective methods are needed for monitoring water for emerging viruses. Combining water filtration with molecular methods such as PCR is a fast and effective way for detecting viruses. The objective of this study was to apply a convenient method for the detection of AIV in natural water samples. Distilled water and lake, river, and seawater were artificially contaminated with AIV (H5N3) and passed through a filter system. AIV was detected from filter membrane by real-time RT-PCR. The performance of Zetapor, SMWP, and Sartobind D5F membranes in recovering influenza viruses was first evaluated using contaminated distilled water. SWMP, which gave the highest virus recoveries, was then compared with a pre-filter combined GF/F filter membrane in a trial using natural water samples. In this study, the cellulose membrane SMWP was found to be practical for recovery of AIVs in water. Viral yields varied between 62.1 and 65.9% in distilled water and between 1 and 16.7% in natural water samples. The borosilicate glass membrane GF/F combined with pre-filter was also feasible in filtering natural water samples with viral yields from 1.98 to 7.33%. The methods described can be used for monitoring fresh and seawater samples for the presence of AIV and to determine the source of AIV transmission in an outbreak situation.