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Use of Oxidative Stress Responses to Determine the Efficacy of Inactivation Treatments on Cryptosporidium Oocysts

1
Laboratory of Parasitology, Department of Paraclinical Sciences, Faculty of Veterinary Medicine, Norwegian University of Life Sciences, Oluf Thesens vei 22, 1433 Ås, Norway
2
NABAS AS, Moer Allé 33, 1435 Ås, Norway
*
Author to whom correspondence should be addressed.
Academic Editor: Eric Viscogliosi
Microorganisms 2021, 9(7), 1463; https://doi.org/10.3390/microorganisms9071463
Received: 16 June 2021 / Revised: 4 July 2021 / Accepted: 5 July 2021 / Published: 8 July 2021
(This article belongs to the Section Parasitology)
Cryptosporidium oocysts are known for being very robust, and their prolonged survival in the environment has resulted in outbreaks of cryptosporidiosis associated with the consumption of contaminated water or food. Although inactivation methods used for drinking water treatment, such as UV irradiation, can inactivate Cryptosporidium oocysts, they are not necessarily suitable for use with other environmental matrices, such as food. In order to identify alternative ways to inactivate Cryptosporidium oocysts, improved methods for viability assessment are needed. Here we describe a proof of concept for a novel approach for determining how effective inactivation treatments are at killing pathogens, such as the parasite Cryptosporidium. RNA sequencing was used to identify potential up-regulated target genes induced by oxidative stress, and a reverse transcription quantitative PCR (RT-qPCR) protocol was developed to assess their up-regulation following exposure to different induction treatments. Accordingly, RT-qPCR protocols targeting thioredoxin and Cryptosporidium oocyst wall protein 7 (COWP7) genes were evaluated on mixtures of viable and inactivated oocysts, and on oocysts subjected to various potential inactivation treatments such as freezing and chlorination. The results from the present proof-of-concept experiments indicate that this could be a useful tool in efforts towards assessing potential technologies for inactivating Cryptosporidium in different environmental matrices. Furthermore, this approach could also be used for similar investigations with other pathogens. View Full-Text
Keywords: Cryptosporidium; gene expression; oocyst; viability; RNA-Seq; RT-qPCR Cryptosporidium; gene expression; oocyst; viability; RNA-Seq; RT-qPCR
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MDPI and ACS Style

Temesgen, T.T.; Tysnes, K.R.; Robertson, L.J. Use of Oxidative Stress Responses to Determine the Efficacy of Inactivation Treatments on Cryptosporidium Oocysts. Microorganisms 2021, 9, 1463. https://doi.org/10.3390/microorganisms9071463

AMA Style

Temesgen TT, Tysnes KR, Robertson LJ. Use of Oxidative Stress Responses to Determine the Efficacy of Inactivation Treatments on Cryptosporidium Oocysts. Microorganisms. 2021; 9(7):1463. https://doi.org/10.3390/microorganisms9071463

Chicago/Turabian Style

Temesgen, Tamirat T., Kristoffer R. Tysnes, and Lucy J. Robertson 2021. "Use of Oxidative Stress Responses to Determine the Efficacy of Inactivation Treatments on Cryptosporidium Oocysts" Microorganisms 9, no. 7: 1463. https://doi.org/10.3390/microorganisms9071463

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