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Synergism in sequential inactivation of Cryptosporidium parvum with trypsin and UV irradiation

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Abstract

Cryptosporidium, a protozoan parasite, in wastewater presents a major public health concern for water safety. However, bactericidal efficiencies of conventional disinfection methods towards Cryptosporidium oocysts are still hampered owing to the presence of their thick outer wall. In this study, we present a novel UV inactivation process where the efficiency has been significantly enhanced by addition of a trypsin pretreatment stage. Notably, inactivation (log-reduction) of oocysts was noted to be 73.75–294.72% higher than that obtained by UV irradiation alone, under identical conditions. Experimental observations and supporting mechanistic analyses suggest that trypsin led to cleavage of the protein layers on the oocyst wall, facilitating penetration of UV radiation into the oocysts leading to degradation of their genomic DNA (gDNA). The dissociative effect of trypsin on the oocyst wall was indicated by the fact that 64.50% of oocysts displayed early apoptosis after trypsinization. Imaging by scanning electron microscopy indicated that this combined treatment led to substantial disruption of the oocyst coat, deforming their shape. This resulted in the release of cellular proteins and gDNA, their concentrations in bulk solution increasing by 1.22–8.60 times. As UV irradiation time was prolonged, gDNA was degraded into smaller fragments with lower molecular masses. Both laddering and diffuse smear patterns in gel analysis indicated significantly detrimental effects on gDNA and viability of oocysts. Overall, this study demonstrated enhancement of UV inactivation of Cryptosporidium oocysts by trypsin and explored the underlying mechanisms for the process.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was funded by the National Natural Science Foundation of China (Nos. 51908062 and 51978135). It was also supported by the Scientific and Technological Development Plan Project of Changchun City (No. 21ZY30) and Jilin Province (No. 20200201042JC), and Industrial Technology Research and Development Plan of Jilin Provincial Development and Reform Commission (No. 2020C033-5).

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Authors and Affiliations

  1. Jilin Academy of Agricultural Sciences, Changchun, 130033, China

    Dan Xiao & Siyue Wang

  2. Jilin Academy of Animal Husbandry and Veterinary Medicine, Changchun, 130062, China

    Nan Wang

  3. School of Chemical Engineering, Changchun University of Technology, Changchun, 130012, China

    Shiheng Chen

  4. School of Environment, Northeast Normal University, Changchun, 130117, China

    Xiangyi Yuan, Wei Fan & Mingxin Huo

Authors
  1. Dan Xiao
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  2. Nan Wang
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  3. Shiheng Chen
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  4. Siyue Wang
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  5. Xiangyi Yuan
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  6. Wei Fan
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  7. Mingxin Huo
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Contributions

Dan Xaio and Wei Fan conceptualized, designed the experiments and wrote the manuscript. Dan Xaio, Nan Wang, Shiheng Chen, and Siyue Wang conducted the experiments and analyzed the data. Xiangyi Yuan and Mingxin Huo discussed the results and improved the manuscript. All the authors listed have made a substantial contribution to the work, and approved it for publication.

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Correspondence to Wei Fan.

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Xiao, D., Wang, N., Chen, S. et al. Synergism in sequential inactivation of Cryptosporidium parvum with trypsin and UV irradiation. Environ Sci Pollut Res 30, 8354–8362 (2023). https://doi.org/10.1007/s11356-022-24408-4

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