Identification of eukaryotic microorganisms with 18S rRNA next-generation sequencing in wastewater treatment plants, with a more targeted NGS approach required for Cryptosporidium detection
While some microbial eukaryotes can improve effluent quality in wastewater treatment plants (WWTPs), eukaryotic waterborne pathogens are a threat to public health. This study aimed to identify Eukarya, particularly faecal pathogens including Cryptosporidium, in different treatment stages (influent,...
| Main Authors: | , , , , , |
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| Format: | Journal Article |
| Language: | English |
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PERGAMON-ELSEVIER SCIENCE LTD
2019
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| Online Access: | http://purl.org/au-research/grants/arc/LP130100602 http://hdl.handle.net/20.500.11937/90970 |
| _version_ | 1848765475450781696 |
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| author | Zahedi, A. Greay, T.L. Paparini, A. Linge, Kathryn Joll, Cynthia Ryan, U.M. |
| author_facet | Zahedi, A. Greay, T.L. Paparini, A. Linge, Kathryn Joll, Cynthia Ryan, U.M. |
| author_sort | Zahedi, A. |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | While some microbial eukaryotes can improve effluent quality in wastewater treatment plants (WWTPs), eukaryotic waterborne pathogens are a threat to public health. This study aimed to identify Eukarya, particularly faecal pathogens including Cryptosporidium, in different treatment stages (influent, intermediate and effluent) from four WWTPs in Western Australia (WA). Three WWTPs that utilise stabilisation ponds and one WWTP that uses activated sludge (oxidation ditch) treatment technologies were sampled. Eukaryotic 18S rRNA (18S) was targeted in the wastewater samples (n = 26) for next-generation sequencing (NGS), and a mammalian-blocking primer was used to reduce the amplification of mammalian DNA. Overall, bioinformatics analyses revealed 49 eukaryotic phyla in WWTP samples, and three of these phyla contained human intestinal parasites, which were primarily detected in the influent. These human intestinal parasites either had a low percent sequence composition or were not detected in the intermediate and effluent stages and included the amoebozoans Endolimax sp., Entamoeba sp. and Iodamoeba sp., the human pinworm Enterobius vermicularis (Nematoda), and Blastocystis sp. subtypes (Sarcomastigophora). Six Blastocystis subtypes and four Entamoeba species were identified by eukaryotic 18S NGS, however, Cryptosporidium sp. and Giardia sp. were not detected. Real-time polymerase chain reaction (PCR) also failed to detect Giardia, but Cryptosporidium-specific NGS detected Cryptosporidium in all WWTPs, and a total of nine species were identified, including five zoonotic pathogens. Although eukaryotic 18S NGS was able to identify some faecal pathogens, this study has demonstrated that more specific NGS approaches for pathogen detection are more sensitive and should be applied to future wastewater pathogen assessments. |
| first_indexed | 2025-11-14T11:35:50Z |
| format | Journal Article |
| id | curtin-20.500.11937-90970 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T11:35:50Z |
| publishDate | 2019 |
| publisher | PERGAMON-ELSEVIER SCIENCE LTD |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-909702023-05-03T07:30:21Z Identification of eukaryotic microorganisms with 18S rRNA next-generation sequencing in wastewater treatment plants, with a more targeted NGS approach required for Cryptosporidium detection Zahedi, A. Greay, T.L. Paparini, A. Linge, Kathryn Joll, Cynthia Ryan, U.M. Science & Technology Technology Life Sciences & Biomedicine Physical Sciences Engineering, Environmental Environmental Sciences Water Resources Engineering Environmental Sciences & Ecology Wastewater Cryptosporidium Blastotystis Entamoeba Next-generation sequencing ZOONOTIC CRYPTOSPORIDIUM ENTAMOEBA-MOSHKOVSKII BLASTOCYSTIS SUBTYPES GENETIC DIVERSITY GIARDIA-CYSTS PROTOZOA PREVALENCE GENOTYPES HOMINIS PARVUM Blastocystis Cryptosporidium Entamoeba Next-generation sequencing Wastewater Animals Cryptosporidium Eukaryota Feces High-Throughput Nucleotide Sequencing Humans RNA, Ribosomal, 18S Wastewater Western Australia Feces Animals Humans Cryptosporidium RNA, Ribosomal, 18S Western Australia Eukaryota High-Throughput Nucleotide Sequencing Wastewater While some microbial eukaryotes can improve effluent quality in wastewater treatment plants (WWTPs), eukaryotic waterborne pathogens are a threat to public health. This study aimed to identify Eukarya, particularly faecal pathogens including Cryptosporidium, in different treatment stages (influent, intermediate and effluent) from four WWTPs in Western Australia (WA). Three WWTPs that utilise stabilisation ponds and one WWTP that uses activated sludge (oxidation ditch) treatment technologies were sampled. Eukaryotic 18S rRNA (18S) was targeted in the wastewater samples (n = 26) for next-generation sequencing (NGS), and a mammalian-blocking primer was used to reduce the amplification of mammalian DNA. Overall, bioinformatics analyses revealed 49 eukaryotic phyla in WWTP samples, and three of these phyla contained human intestinal parasites, which were primarily detected in the influent. These human intestinal parasites either had a low percent sequence composition or were not detected in the intermediate and effluent stages and included the amoebozoans Endolimax sp., Entamoeba sp. and Iodamoeba sp., the human pinworm Enterobius vermicularis (Nematoda), and Blastocystis sp. subtypes (Sarcomastigophora). Six Blastocystis subtypes and four Entamoeba species were identified by eukaryotic 18S NGS, however, Cryptosporidium sp. and Giardia sp. were not detected. Real-time polymerase chain reaction (PCR) also failed to detect Giardia, but Cryptosporidium-specific NGS detected Cryptosporidium in all WWTPs, and a total of nine species were identified, including five zoonotic pathogens. Although eukaryotic 18S NGS was able to identify some faecal pathogens, this study has demonstrated that more specific NGS approaches for pathogen detection are more sensitive and should be applied to future wastewater pathogen assessments. 2019 Journal Article http://hdl.handle.net/20.500.11937/90970 10.1016/j.watres.2019.04.041 English http://purl.org/au-research/grants/arc/LP130100602 http://creativecommons.org/licenses/by-nc-nd/4.0/ PERGAMON-ELSEVIER SCIENCE LTD fulltext |
| spellingShingle | Science & Technology Technology Life Sciences & Biomedicine Physical Sciences Engineering, Environmental Environmental Sciences Water Resources Engineering Environmental Sciences & Ecology Wastewater Cryptosporidium Blastotystis Entamoeba Next-generation sequencing ZOONOTIC CRYPTOSPORIDIUM ENTAMOEBA-MOSHKOVSKII BLASTOCYSTIS SUBTYPES GENETIC DIVERSITY GIARDIA-CYSTS PROTOZOA PREVALENCE GENOTYPES HOMINIS PARVUM Blastocystis Cryptosporidium Entamoeba Next-generation sequencing Wastewater Animals Cryptosporidium Eukaryota Feces High-Throughput Nucleotide Sequencing Humans RNA, Ribosomal, 18S Wastewater Western Australia Feces Animals Humans Cryptosporidium RNA, Ribosomal, 18S Western Australia Eukaryota High-Throughput Nucleotide Sequencing Wastewater Zahedi, A. Greay, T.L. Paparini, A. Linge, Kathryn Joll, Cynthia Ryan, U.M. Identification of eukaryotic microorganisms with 18S rRNA next-generation sequencing in wastewater treatment plants, with a more targeted NGS approach required for Cryptosporidium detection |
| title | Identification of eukaryotic microorganisms with 18S rRNA next-generation sequencing in wastewater treatment plants, with a more targeted NGS approach required for Cryptosporidium detection |
| title_full | Identification of eukaryotic microorganisms with 18S rRNA next-generation sequencing in wastewater treatment plants, with a more targeted NGS approach required for Cryptosporidium detection |
| title_fullStr | Identification of eukaryotic microorganisms with 18S rRNA next-generation sequencing in wastewater treatment plants, with a more targeted NGS approach required for Cryptosporidium detection |
| title_full_unstemmed | Identification of eukaryotic microorganisms with 18S rRNA next-generation sequencing in wastewater treatment plants, with a more targeted NGS approach required for Cryptosporidium detection |
| title_short | Identification of eukaryotic microorganisms with 18S rRNA next-generation sequencing in wastewater treatment plants, with a more targeted NGS approach required for Cryptosporidium detection |
| title_sort | identification of eukaryotic microorganisms with 18s rrna next-generation sequencing in wastewater treatment plants, with a more targeted ngs approach required for cryptosporidium detection |
| topic | Science & Technology Technology Life Sciences & Biomedicine Physical Sciences Engineering, Environmental Environmental Sciences Water Resources Engineering Environmental Sciences & Ecology Wastewater Cryptosporidium Blastotystis Entamoeba Next-generation sequencing ZOONOTIC CRYPTOSPORIDIUM ENTAMOEBA-MOSHKOVSKII BLASTOCYSTIS SUBTYPES GENETIC DIVERSITY GIARDIA-CYSTS PROTOZOA PREVALENCE GENOTYPES HOMINIS PARVUM Blastocystis Cryptosporidium Entamoeba Next-generation sequencing Wastewater Animals Cryptosporidium Eukaryota Feces High-Throughput Nucleotide Sequencing Humans RNA, Ribosomal, 18S Wastewater Western Australia Feces Animals Humans Cryptosporidium RNA, Ribosomal, 18S Western Australia Eukaryota High-Throughput Nucleotide Sequencing Wastewater |
| url | http://purl.org/au-research/grants/arc/LP130100602 http://hdl.handle.net/20.500.11937/90970 |