Filtration extraction method using microfluidic channel for measuring environmental DNA

The environmental DNA (eDNA) method, which is widely applied for biomonitoring, is limited to laboratory analysis and processing. In this study, we developed a filtration/extraction component using a microfluidic channel, Biryu-Chip (BC), and a filtration/extraction method, BC method, to minimize the volume of the sample necessary for DNA extraction and subsequent PCR amplification. We tested the performance of the BC method and compared it with the Sterivex filtration/extraction method using aquarium and river water samples. We observed that using the BC method, the same concentration of the extracted DNA was obtained with 1/20-1/40 of the filtration volume of the Sterivex method, suggesting that the BC method can be widely used for eDNA measurement. In addition, we could perform on-site measurements of eDNA within 30 min using a mobile PCR device. Using the BC method, filtration and extraction could be performed easily and quickly. The PCR results obtained by the BC method were similar to those obtained by the Sterivex method. However, the BC method required fewer steps and therefore, the risk of DNA contamination could be reduced. When combined with a mobile PCR, the BC method can be applied to easily detect eDNA within 30 min from 10 mL of the water sample, even on-site.

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Filtration extraction method using microfluidic channel for measuring environmental DNA

10.22541/au.163854179.99663512/v1 ◽

2021 ◽

Author(s):

Takashi Fukuzawa ◽

Yuichi Kameda ◽

Hisao Nagata ◽

Naofumi Nishizawa ◽

Hideyuki Doi

Keyword(s):

Water Sample ◽

River Water ◽

Dna Extraction ◽

Water Samples ◽

Extraction Method ◽

Microfluidic Channel ◽

Pcr Amplification ◽

Environmental Dna ◽

Laboratory Analysis ◽

River Water Samples

The environmental DNA (eDNA) method, which is widely applied for biomonitoring, is limited to laboratory analysis and processing. In this study, we developed a filtration/extraction component using a microfluidic channel, Biryu-Chip (BC), and a filtration/extraction method, BC method, to minimize the volume of the sample necessary for DNA extraction and subsequent PCR amplification. We tested the performance of the BC method and compared it with the Sterivex filtration/extraction method using aquarium and river water samples. We observed that using the BC method, the same concentration of the extracted DNA was obtained with 1/20–1/40 of the filtration volume of the Sterivex method, suggesting that the BC method can be widely used for eDNA measurement. In addition, we could perform on-site measurements of eDNA within 30 min using a mobile PCR device. Using the BC method, filtration and extraction could be performed easily and quickly. The PCR results obtained by the BC method were similar to those obtained by the Sterivex method. The BC method required fewer steps and therefore, the risk of DNA contamination could be reduced. When combined with a mobile PCR, the BC method can be applied to easily detect eDNA within 30 min from a few 10 mL of the water sample, even on-site.

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First environmental surveillance for the presence of SARS-CoV-2 RNA in wastewater and river water in Japan

10.1101/2020.06.04.20122747 ◽

2020 ◽

Cited By ~ 6

Author(s):

Eiji Haramoto ◽

Bikash Malla ◽

Ocean Thakali ◽

Masaaki Kitajima

Keyword(s):

River Water ◽

Water Samples ◽

Extraction Method ◽

Limit Of Detection ◽

Rna Extraction ◽

Treatment Plant ◽

Treated Wastewater ◽

Wastewater Sample ◽

River Water Samples ◽

Wastewater Samples

ABSTRACTWastewater-based epidemiology is a powerful tool to understand the actual incidence of coronavirus disease 2019 (COVID-19) in a community because severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent of COVID-19, can be shed in the feces of infected individuals regardless of their symptoms. The present study aimed to assess the presence of SARS-CoV-2 RNA in wastewater and river water in Yamanashi Prefecture, Japan, using four quantitative and two nested PCR assays. Influent and secondary-treated (before chlorination) wastewater samples and river water samples were collected five times from a wastewater treatment plant and three times from a river, respectively, between March 17 and May 7, 2020. The wastewater and river water samples (200–5,000 mL) were processed by using two different methods: the electronegative membrane-vortex (EMV) method and the membrane adsorption-direct RNA extraction method. Based on the observed concentrations of indigenous pepper mild mottle virus RNA, the EMV method was found superior to the membrane adsorption-direct RNA extraction method. SARS-CoV-2 RNA was successfully detected in one of five secondary-treated wastewater samples with a concentration of 2.4 × 103 copies/L by N_Sarbeco qPCR assay following the EMV method, whereas all the influent samples were tested negative for SARS-CoV-2 RNA. This result could be attributed to higher limit of detection for influent (4.0 × 103–8.2 × 104 copies/L) with a lower filtration volume (200 mL) compared to that for secondary-treated wastewater (1.4 × 102–2.5 × 103 copies/L) with a higher filtration volume of 5,000 mL. None of the river water samples tested positive for SARS-CoV-2 RNA. Comparison with the reported COVID-19 cases in Yamanashi Prefecture showed that SARS-CoV-2 RNA was detected in the secondary-treated wastewater sample when the cases peaked in the community. This is the first study reporting the detection of SARS-CoV-2 RNA in wastewater in Japan.

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Faculty Opinions recommendation of Conjugative transfer of the IncP-7 carbazole degradative plasmid, pCAR1, in river water samples.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1091365.545802 ◽

2007 ◽

Author(s):

Peter Williams

Keyword(s):

River Water ◽

Water Samples ◽

Conjugative Transfer ◽

River Water Samples ◽

Degradative Plasmid

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Fabrication of dendritic nanoporous gold via a two-step amperometric approach: Application for electrochemical detection of methyl parathion in river water samples

Talanta ◽

10.1016/j.talanta.2021.122130 ◽

2021 ◽

Vol 226 ◽

pp. 122130

Author(s):

Gilberto J. Silva Junior ◽

Jéssica Soares Guimarães Selva ◽

Anandhakumar Sukeri ◽

Josué M. Gonçalves ◽

Matias Regiart ◽

...

Keyword(s):

River Water ◽

Electrochemical Detection ◽

Water Samples ◽

Methyl Parathion ◽

Nanoporous Gold ◽

River Water Samples

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Modified paramagnetic beads in a microfluidic system for the determination of ethinylestradiol (EE2) in river water samples

Biosensors and Bioelectronics ◽

10.1016/j.bios.2009.10.031 ◽

2010 ◽

Vol 25 (6) ◽

pp. 1376-1381 ◽

Cited By ~ 48

Author(s):

Noelia A. Martínez ◽

Rudolf J. Schneider ◽

Germán A. Messina ◽

Julio Raba

Keyword(s):

River Water ◽

Water Samples ◽

Microfluidic System ◽

River Water Samples ◽

Paramagnetic Beads

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Coagulant potentials of Moringa oleifera seeds in water purification

FLORA AND FAUNA ◽

10.33451/florafauna.v26i2pp183-189 ◽

2020 ◽

Vol 26 (2) ◽

Author(s):

R. V. Thorat ◽

D. A. Malvekar ◽

A. U. Sutar

Keyword(s):

Water Sample ◽

River Water ◽

Water Purification ◽

Water Samples ◽

Moringa Oleifera ◽

Sewage Water ◽

Chemical Parameters ◽

Natural Coagulant ◽

Physico Chemical ◽

Almost All

The main objective of this study is to confirm the potentials of Moringa oleifera seeds over alum for water purification. Various doses of Moringa seed powder 4, 8, 12 g/l were taken and checked for treatment of river and sewage water. After treatment of seed powder with water samples were analyzed for physico-chemical parameters like pH, Absorbance, TDS, TSS, Hardness, Chlorides, Conductivity, Turbidity, MPN and DO. Almost all parameters showed reduction with increasing dosage of Moringa seed powder. Similar doses of alum were checked with river water sample and were analyzed for above mentioned parameters. The results obtained showed that seed powder (natural coagulant) is more effective than conventional chemical coagulant., alum. The seed of Moringa oleifera is cheap, eco-friendly and non-toxic, can be effectively used as a coagulant for river and sewage water purification.

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