Development of a centrifugation-based method for the qualitative molecular detection of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in surface water samples.
- 1Department of Veterinary Medicine and Animal Sciences, Università degli Studi di Milano, Lodi, Italy.
- 2Research Laboratory of Animal Parasitic Diseases and Zoonoses (ParVetLab), Università degli Studi di Milano, Lodi, Italy.
- 3National Research Council of Italy, Water Research Institute (CNR-IRSA), Brugherio Unit, Brugherio, Italy.
Abstract
Workflow for waterborne parasites DNA detection in surface water. Screening for environmental surveillance in monitoring and resource-limited settings. Zoonotic protozoa and microsporidia are waterborne pathogens with implications for animal and human health. Their resistant stages can persist in freshwater. Standard reference methods for water analysis are often limited to specific parasites, require large water volumes, and depends on expensive equipment. Practical alternatives are needed to facilitate parasite detection in water. This study aimed to develop a pellet-based workflow to allow the DNA detection of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi from the same surface water sample. Three organism-specific spiked controls and eight environmental samples from four freshwater sites, collected over two months, were processed by centrifugation to concentrate the pellet fraction, while the supernatant recovered post-centrifugation was filtered through membranes. The samples were collected as part of a broader ongoing year-long monitoring project. DNA was extracted from both fractions and analysed by qualitative PCR assays. Pellets yielded positive results whereas no positivity was detected in filter membranes, suggesting pellet fractions as the most suitable for downstream molecular analysis. This qualitative pellet-based processing approach was designed to provide an assessment and a practical screening strategy for environmental surveillance.
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BibTeX
@article{sechi2026development,
title = {{Development of a centrifugation-based method for the qualitative molecular detection of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in surface water samples.}},
author = {Susanna Adelaide Sechi and Chiara Stocchero and Carolina Allievi and Diego Copetti and Fabrizio Stefani and Lucia Valsecchi and Alessandra Cafiso and Chiara Bazzocchi and Maria Teresa Manfredi},
journal = {MethodsX},
year = {2026},
month = oct,
doi = {10.1016/j.mex.2026.104195},
url = {https://europepmc.org/article/MED/42903624},
note = {CC BY},
}
RIS
TY - JOUR
TI - Development of a centrifugation-based method for the qualitative molecular detection of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in surface water samples.
AU - Susanna Adelaide Sechi
AU - Chiara Stocchero
AU - Carolina Allievi
AU - Diego Copetti
AU - Fabrizio Stefani
AU - Lucia Valsecchi
AU - Alessandra Cafiso
AU - Chiara Bazzocchi
AU - Maria Teresa Manfredi
PY - 2026
DA - 2026/10/08
JO - MethodsX
DO - 10.1016/j.mex.2026.104195
UR - https://europepmc.org/article/MED/42903624
AB - Workflow for waterborne parasites DNA detection in surface water. Screening for environmental surveillance in monitoring and resource-limited settings. Zoonotic protozoa and microsporidia are waterborne pathogens with implications for animal and human health. Their resistant stages can persist in freshwater. Standard reference methods for water analysis are often limited to specific parasites, require large water volumes, and depends on expensive equipment. Practical alternatives are needed to facilitate parasite detection in water. This study aimed to develop a pellet-based workflow to allow the DNA detection of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi from the same surface water sample. Three organism-specific spiked controls and eight environmental samples from four freshwater sites, collected over two months, were processed by centrifugation to concentrate the pellet fraction, while the supernatant recovered post-centrifugation was filtered through membranes. The samples were collected as part of a broader ongoing year-long monitoring project. DNA was extracted from both fractions and analysed by qualitative PCR assays. Pellets yielded positive results whereas no positivity was detected in filter membranes, suggesting pellet fractions as the most suitable for downstream molecular analysis. This qualitative pellet-based processing approach was designed to provide an assessment and a practical screening strategy for environmental surveillance.
SN - 2215-0161
ER -