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Frontiers of Chemical Science and Engineering

Front. Chem. Sci. Eng.    2019, Vol. 13 Issue (3) : 608-615
Cryptosporidium parvum oocyst directed assembly of gold nanoparticles and graphene oxide
Sona Jain1, Zhicheng Huang1, Brent R. Dixon2, Syed Sattar3, Juewen Liu1()
1. Department of Chemistry, Waterloo Institute for Nanotechnology, University of Waterloo, Ontario, N2L 3G1, Canada
2. Bureau of Microbial Hazards, Food Directorate, Health Canada, Ontario, K1A 0K9, Canada
3. Faculty of Medicine, University of Ottawa, Ontario, K1H 8M5, Canada
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Understanding the interactions between inorganic nanomaterials and biological species is an important topic for surface and environmental chemistry. In this work, we systematically studied the oocysts of Cryptosporidium parvum as a model protozoan parasite and its interaction with gold nanoparticles (AuNPs) and graphene oxide (GO). The as-prepared citrate-capped AuNPs adsorb strongly on the oocysts leading to a vivid color change. The adsorption of the AuNPs was confirmed by transmission electron microscopy. Heat treatment fully inhibited the color change, indicating a large change of surface chemistry of the oocysts that can be probed by the AuNPs. Adding proteases such as trypsin and proteinase K partially inhibited the color change. DNA-capped AuNPs, on the other hand, could not be adsorbed by the oocysts. GO was found to wrap around the oocysts forming a conformal shell reflecting the shape of the oocysts. Both citrate-capped AuNPs and GO compromised the membrane integrity of the oocysts as indicated by the propidium iodide staining experiment, and they may be potentially used for inactivating the oocysts. This is the first example of using nanomaterials to probe the surface of the oocysts, and it suggests the possibility of using such organisms to template the assembly of nanomaterials.

Keywords nanomaterials      toxicology      water      biosensors     
Corresponding Authors: Juewen Liu   
Just Accepted Date: 27 March 2019   Online First Date: 29 April 2019    Issue Date: 22 August 2019
 Cite this article:   
Sona Jain,Zhicheng Huang,Brent R. Dixon, et al. Cryptosporidium parvum oocyst directed assembly of gold nanoparticles and graphene oxide[J]. Front. Chem. Sci. Eng., 2019, 13(3): 608-615.
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Sona Jain
Zhicheng Huang
Brent R. Dixon
Syed Sattar
Juewen Liu
Fig.1  (a) A schematic representation of the multilayered oocyst wall structure. The approximate thickness of each layer is marked on the left. (b) An optical micrograph (wet mount) of the oocysts used in this work
Fig.2  TEM micrographs of the (a) AuNPs and (b) GO used in this work
Fig.3  (a) Visual inspection of the interaction of C. parvum oocysts with citrate-capped 13 nm AuNP (Tube 1: AuNPs dispersed in the salt-containing buffer for storing the oocysts; Tube 2: AuNPs dispersed in the supernatant after second wash using water; Tube 3: AuNPs mixed with the oocysts after second wash with water; Tube 4: 13 nm citrate-capped AuNPs without any treatment); (b) Wet mount optical micrograph of the oocysts treated with AuNP; (c) A TEM micrograph of the oocysts mixed with AuNPs and fixed with formaldehyde; (d) A higher resolution TEM micrograph of the same sample
Fig.4  (a) Schematic representation of the preparation of DNA-capped AuNP by using a thiolated DNA (SH-DNA); (b) Photographs of oocysts treated with citrate and DNA-capped AuNPs; (c) TEM of the oocysts plus DNA-capped AuNPs (The very small black dots throughout the micrograph are DNA-capped AuNPs)
Fig.5  (a) Visual inspection of the effect of heat and trypsin treatment on the interaction of oocysts with citrate-capped AuNPs (Tube 1: AuNPs mixed with untreated oocysts (without heat/trypsin treatment); Tube 2: AuNP control; Tube 3: With heat-treated oocysts (10 min); Tube 4: With trypsin treated oocysts (1 h)); (b) UV-vis absorption spectra of untreated and heat-treated oocysts when incubated with citrate-capped AuNP; (c) UV-vis spectra for oocysts treated with proteinase K, trypsin-HCl and trypsin-EDTA before incubation with citrate-capped AuNP
Fig.6  TEM micrographs of C. parvum oocyst interaction with GO. (a,b): Without fixing and (c,d) fixed with formaldehyde. The capsule features are likely templated by the oocysts
Fig.7  C. parvum oocysts stained with PI (Micrographs were taken using a confocal microscope). (a) Untreated oocysts; (b) heat-treated oocysts at 75°C for 10 min; (c) oocysts treated with AuNP; (d) oocysts treated with GO
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