Takahiro Sekikawa

A simple method for extracting DNA from Cryptosporidium oocysts using the anionic surfactant LSS

Detection of low amounts of Cryptosporidium oocysts in raw water sources is considered an important component in the management, prevention and control of Cryptosporidium in drinking water supplies as Cryptosporidium causes massive waterborne outbreaks worldwide. As Cryptosporidium has a robust oocyst that is extremely resistant to chlorine and other drinking water disinfectants, both the freeze-thaw method and DNA extraction kits have been commonly used for extracting and purifying DNA from the oocyst. However, the DNA extraction procedures are time consuming and costly. Therefore, a simple and low-cost method to extract and purify DNA from the robust oocyst has been required. In this study, we discussed a simple method for detecting Cryptosporidium DNA with the anionic surfactant, n-lauroylsarcosine sodium salt (LSS) using the loop-mediated isothermal amplification (LAMP) to eliminate the need for the freeze-thaw method and the DNA extraction kits. As a result, Bst DNA polymerase was inhibited by 0.1% LSS but not 0.01% LSS and 5% Triton X-100 or Tween 20. Although DNA was extracted from the oocysts by incubating with 0.1% LSS at 90°C for 15 min, Bst DNA polymerase was inhibited by 0.1% LSS. The inhibition by 0.1% LSS was suppressed by adding 5% of the nonionic surfactants, Triton X-100 or Tween 20. The concentration of LSS in a LAMP tube was 0.01% while that in an incubation tube was 0.1%, because LSS in an incubation tube was diluted by a factor of 10 at the DNA amplification process. Therefore, we found that ten oocysts of Cryptosporidium parvum could be detected by incubation with 0.1% LSS, without removing LSS or adding the nonionic surfactants in the LAMP method.

Effects of introducing energy recovery processes to the municipal solid waste management system in Ulaanbaatar, Mongolia

Currently, most developing countries have not set up municipal solid waste management systems with a view of recovering energy from waste or reducing greenhouse gas emissions. In this article, we have studied the possible effects of introducing three energy recovery processes either as a single or combination approach, refuse derived fuel production, incineration and waste power generation, and methane gas recovery from landfill and power generation in Ulaanbaatar, Mongolia, as a case study. We concluded that incineration process is the most suitable as first introduction of energy recovery. To operate it efficiently, 3Rs strategies need to be promoted. And then, RDF production which is made of waste papers and plastics in high level of sorting may be considered as the second step of energy recovery. However, safety control and marketability of RDF will be required at that moment.