Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully develop...Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously,namely A lexandrium tamarense,Gyrodinium instriatum,Heterosigma akashiwo,Karenia mikimotoi,Prorocentrum donghaiense,Prorocentrum minimum,Ulva compressa,Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection(LOD) of 0.5 ng of genomic DNA(orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether,230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%,respectively,relative to conventional morphological methods. This indicated that this high-throughput,automatic,and specific method is well suited for the detection of algae in water samples.展开更多
UMhlathuze River is the main recipient of domestic sewage, industrial as well as agricultural waste from local industrial and agricultural practices. It is the effluent ＂dumping sites＂ for most human activity practi...UMhlathuze River is the main recipient of domestic sewage, industrial as well as agricultural waste from local industrial and agricultural practices. It is the effluent ＂dumping sites＂ for most human activity practices. Contamination of UMhlathuze River by agricultural and industrial developments makes communities around UMhlathuze area susceptible to potential outbreaks of water-borne illnesses as well as the risk of ingesting carcinogenic substances. In establishment of the effect of human developments on the UMhlathuze River, four different sites representing different human activities were used for sampling along the river. Temperature and pH were monitored in siitu and were found to vary between 19-21℃and 6.2-7.8, respectively. Chemical oxygen demand （COD） was found to be higher in areas affected by both industrial and agricultural activities. Spectroquadrant Pharo 300 （Merck） was used for chemical analysis of water. Agricultural waste recipient area had high concentration of phosphate, ammonia and nitrate. Parts of the river receiving effluent from treated wastewater had high sulphide, nitrate and ammonia concentrations. Industrial areas had high concentration of heavy metals （e.g., aluminium）. Microorganisms found in all sites included Escherichia coli and species of Salmonella, Shigella, Citrobacter, Serratia and Enterobacter, although their quantities differed from agricultural to industrial sites. There was a positive correlation between contamination levels and human activities which ascribe river pollution to activities around the area. Agricultural and industrial development activities practiced around UMhlathuze River have a huge contribution to the continued deterioration of the quality of water at UMhlathuze River.展开更多
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA092001)the Science and Technology Project of Zhejiang Province(No.2013C03045-1)+2 种基金the Zhejiang Marine Biotechnology Innovation Team(No.2010R50029-12)the Natural Science Foundation of Ningbo City of China(No.2013A610168)the KC Wong Magna Fund in Ningbo University
文摘Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously,namely A lexandrium tamarense,Gyrodinium instriatum,Heterosigma akashiwo,Karenia mikimotoi,Prorocentrum donghaiense,Prorocentrum minimum,Ulva compressa,Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection(LOD) of 0.5 ng of genomic DNA(orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether,230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%,respectively,relative to conventional morphological methods. This indicated that this high-throughput,automatic,and specific method is well suited for the detection of algae in water samples.
文摘UMhlathuze River is the main recipient of domestic sewage, industrial as well as agricultural waste from local industrial and agricultural practices. It is the effluent ＂dumping sites＂ for most human activity practices. Contamination of UMhlathuze River by agricultural and industrial developments makes communities around UMhlathuze area susceptible to potential outbreaks of water-borne illnesses as well as the risk of ingesting carcinogenic substances. In establishment of the effect of human developments on the UMhlathuze River, four different sites representing different human activities were used for sampling along the river. Temperature and pH were monitored in siitu and were found to vary between 19-21℃and 6.2-7.8, respectively. Chemical oxygen demand （COD） was found to be higher in areas affected by both industrial and agricultural activities. Spectroquadrant Pharo 300 （Merck） was used for chemical analysis of water. Agricultural waste recipient area had high concentration of phosphate, ammonia and nitrate. Parts of the river receiving effluent from treated wastewater had high sulphide, nitrate and ammonia concentrations. Industrial areas had high concentration of heavy metals （e.g., aluminium）. Microorganisms found in all sites included Escherichia coli and species of Salmonella, Shigella, Citrobacter, Serratia and Enterobacter, although their quantities differed from agricultural to industrial sites. There was a positive correlation between contamination levels and human activities which ascribe river pollution to activities around the area. Agricultural and industrial development activities practiced around UMhlathuze River have a huge contribution to the continued deterioration of the quality of water at UMhlathuze River.
