Dehydration of Prions on Environmentally Relevant Surfaces Protects Them from Inactivation by Freezing and Thawing

ABSTRACTChronic wasting disease (CWD) is an emerging prion disease in North America. Recent identification of CWD in wild cervids from Norway raises the concern of the spread of CWD in Europe. CWD infectivity can enter the environment through live animal excreta and carcasses where it can bind to soil. Well-characterized hamster prion strains and CWD field isolates in unadsorbed or soil-adsorbed forms that were either hydrated or dehydrated were subjected to repeated rounds of freezing and thawing. We found that 500 cycles of repeated freezing and thawing of hydrated samples significantly decreased the abundance of PrPScand reduced protein misfolding cyclic amplification (PMCA) seeding activity that could be rescued by binding to soil. Importantly, dehydration prior to freezing and thawing treatment largely protected PrPScfrom degradation, and the samples maintained PMCA seeding activity. We hypothesize that redistribution of water molecules during the freezing and thawing process alters the stability of PrPScaggregates. Overall, these results have significant implications for the assessment of prion persistence in the environment.IMPORTANCEPrions excreted into the environment by infected animals, such as elk and deer infected with chronic wasting disease, persist for years and thus facilitate horizontal transmission of the disease. Understanding the fate of prions in the environment is essential to control prion disease transmission. The significance of our study is that it provides information on the possibility of prion degradation and inactivation under natural weathering processes. This information is significant for remediation of prion-contaminated environments and development of prion disease control strategies.

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Environmental Sources of Scrapie Prions

Journal of Virology ◽

10.1128/jvi.01133-10 ◽

2010 ◽

Vol 84 (21) ◽

pp. 11560-11562 ◽

Cited By ~ 37

Author(s):

Ben C. Maddison ◽

Claire A. Baker ◽

Linda A. Terry ◽

Susan J. Bellworthy ◽

Leigh Thorne ◽

...

Keyword(s):

Environmental Contamination ◽

Disease Transmission ◽

Protein Misfolding ◽

Chronic Wasting Disease ◽

Horizontal Transmission ◽

Wasting Disease ◽

Prion Infectivity ◽

Protein Misfolding Cyclic Amplification ◽

Sheep Farm

ABSTRACT Ovine scrapie and cervine chronic wasting disease show considerable horizontal transmission. Here we report that a scrapie-affected sheep farm has a widespread environmental contamination with prions. Prions were amplified by protein-misfolding cyclic amplification (sPMCA) from seven of nine environmental swab samples taken, including those from metal, plastic, and wooden surfaces. Sheep had been removed from the areas from which the swabs were taken up to 20 days prior to sampling, indicating that prions persist for at least that long. These data implicate inanimate objects as environmental reservoirs for prion infectivity that are likely to contribute to facile disease transmission.

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Generation of human chronic wasting disease in transgenic mice

Acta Neuropathologica Communications ◽

10.1186/s40478-021-01262-y ◽

2021 ◽

Vol 9 (1) ◽

Author(s):

Zerui Wang ◽

Kefeng Qin ◽

Manuel V. Camacho ◽

Ignazio Cali ◽

Jue Yuan ◽

...

Keyword(s):

Transgenic Mice ◽

Prion Disease ◽

Protein Misfolding ◽

Chronic Wasting Disease ◽

Cellular Prion Protein ◽

Spongiform Encephalopathy ◽

Species Barrier ◽

Wasting Disease ◽

Bona Fide

AbstractChronic wasting disease (CWD) is a cervid prion disease caused by the accumulation of an infectious misfolded conformer (PrPSc) of cellular prion protein (PrPC). It has been spreading rapidly in North America and also found in Asia and Europe. Although bovine spongiform encephalopathy (i.e. mad cow disease) is the only animal prion disease known to be zoonotic, the transmissibility of CWD to humans remains uncertain. Here we report the generation of the first CWD-derived infectious human PrPSc by elk CWD PrPSc-seeded conversion of PrPC in normal human brain homogenates using in vitro protein misfolding cyclic amplification (PMCA). Western blotting with human PrP selective antibody confirmed that the PMCA-generated protease-resistant PrPSc was derived from the human PrPC substrate. Two lines of humanized transgenic mice expressing human PrP with either Val or Met at the polymorphic codon 129 developed clinical prion disease following intracerebral inoculation with the PMCA-generated CWD-derived human PrPSc. Diseased mice exhibited distinct PrPSc patterns and neuropathological changes in the brain. Our study, using PMCA and animal bioassays, provides the first evidence that CWD PrPSc can cross the species barrier to convert human PrPC into infectious PrPSc that can produce bona fide prion disease when inoculated into humanized transgenic mice.

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Modified Protein Misfolding Cyclic Amplification Overcomes Real-Time Quaking-Induced Conversion Assay Inhibitors in Deer Saliva To Detect Chronic Wasting Disease Prions

Journal of Clinical Microbiology ◽

10.1128/jcm.00947-18 ◽

2018 ◽

Vol 56 (9) ◽

Cited By ~ 9

Author(s):

Kristen A. Davenport ◽

Clare E. Hoover ◽

Nathaniel D. Denkers ◽

Candace K. Mathiason ◽

Edward A. Hoover

Keyword(s):

Real Time ◽

Protein Misfolding ◽

Chronic Wasting Disease ◽

Horizontal Transmission ◽

Wasting Disease ◽

Protein Misfolding Cyclic Amplification ◽

Rapid Spread ◽

Low Levels ◽

The Republic ◽

The Impact

ABSTRACT Chronic wasting disease (CWD), a fatal neurodegenerative prion disease of cervids, has spread across North America and has been detected in The Republic of Korea, Finland, and Norway. CWD appears to spread by horizontal transmission, and prions shed in saliva, feces, and urine are thought to contribute. However, studies investigating the rapid spread of CWD have been hampered by assay inhibitors and a lack of consistent and sensitive means to detect the relatively low levels of prions in these samples. Here we show that saliva frequently contains an inhibitor of the real-time quaking-induced conversion assay (RT-QuIC) and that the inhibitor is a member of the mucin family. To circumvent the inhibitor, we developed a modified protein misfolding cyclic amplification (PMCA) method to amplify CWD prions in saliva that were undetectable or ambiguous by RT-QuIC. Our results reinforce the impact of saliva in horizontal CWD transmission and highlight the importance of detection optimization.

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Efficient In Vitro Amplification of Chronic Wasting Disease PrPRES

Journal of Virology ◽

10.1128/jvi.00635-07 ◽

2007 ◽

Vol 81 (17) ◽

pp. 9605-9608 ◽

Cited By ~ 71

Author(s):

Timothy D. Kurt ◽

Matthew R. Perrott ◽

Carol J. Wilusz ◽

Jeffrey Wilusz ◽

Surachai Supattapone ◽

...

Keyword(s):

Transgenic Mice ◽

Prion Protein ◽

Protein Misfolding ◽

Chronic Wasting Disease ◽

Body Fluids ◽

The Body ◽

Wasting Disease ◽

Highly Efficient ◽

Significant Step

ABSTRACT Chronic wasting disease (CWD) of cervids is associated with conversion of the normal cervid prion protein, PrPC, to a protease-resistant conformer, PrPCWD. Here we report the use of both nondenaturing amplification and protein-misfolding cyclic amplification (PMCA) to amplify PrPCWD in vitro. Normal brains from deer, transgenic mice expressing cervid PrPC [Tg(cerPrP)1536 mice], and ferrets supported amplification. PMCA using normal Tg(cerPrP)1536 brains as the PrPC substrate produced >6.5 × 109-fold amplification after six rounds. Highly efficient in vitro amplification of PrPCWD is a significant step toward detection of PrPCWD in the body fluids or excreta of CWD-susceptible species.

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Human Prion Disease and Relative Risk Associated with Chronic Wasting Disease

Emerging Infectious Diseases ◽

10.3201/eid1210.060019 ◽

2006 ◽

Vol 12 (10) ◽

pp. 1527-1535 ◽

Cited By ~ 45

Author(s):

Samantha MaWhinney ◽

W. John Pape ◽

Jeri E. Forster ◽

C. Alan Anderson ◽

Patrick Bosque ◽

...

Keyword(s):

Relative Risk ◽

Prion Disease ◽

Chronic Wasting Disease ◽

Human Prion Disease ◽

Wasting Disease

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Failure To Detect Prion Infectivity in Ticks following Prion-Infected Blood Meal

mSphere ◽

10.1128/msphere.00741-20 ◽

2020 ◽

Vol 5 (5) ◽

Author(s):

Ronald A. Shikiya ◽

Anthony E. Kincaid ◽

Jason C. Bartz ◽

Travis J. Bourret

Keyword(s):

Prion Disease ◽

Blood Meal ◽

Red Deer ◽

Chronic Wasting Disease ◽

Mule Deer ◽

Geographic Range ◽

Lymphoid Tissues ◽

Wasting Disease ◽

Prion Infection ◽

Prion Infectivity

ABSTRACT Chronic wasting disease (CWD) is an emerging and fatal contagious prion disease that affects cervids, including mule deer, white-tailed deer, black-tailed deer, red deer reindeer, elk, and moose. CWD prions are widely distributed throughout the bodies of CWD-infected animals and are found in the nervous system, lymphoid tissues, muscle, blood, urine, feces, and antler velvet. The mechanism of CWD transmission in natural settings is unknown. Potential mechanisms of transmission include horizontal, maternal, or environmental routes. Due to the presence of prions in the blood of CWD-infected animals, the potential exists for invertebrates that feed on mammalian blood to contribute to the transmission of CWD. The geographic range of the Rocky Mountain Wood tick, Dermancentor andersoni, overlaps with CWD throughout the northwest United States and southwest Canada, raising the possibility that D. andersoni parasitization of cervids may be involved in CWD transmission. We investigated this possibility by examining the blood meal of D. andersoni that fed upon prion-infected hamsters for the presence of prion infectivity by animal bioassay. None of the hamsters inoculated with a D. andersoni blood meal that had been ingested from prion-infected hamsters developed clinical signs of prion disease or had evidence for a subclinical prion infection. Overall, the data do not demonstrate a role for D. andersoni in the transmission of prion disease. IMPORTANCE Chronic wasting disease (CWD) is an emerging prion disease that affects cervids, including mule deer, white-tailed deer, black-tailed deer, red deer reindeer, elk, and moose. The mechanism of CWD transmission in unknown. Due to the presence of prions in the blood of CWD-infected animals, it is possible for invertebrates that feed on cervid blood to contribute to the transmission of CWD. We examined the blood meal of D. andersoni, a tick with a similar geographic range as cervids, that fed upon prion-infected hamsters for the presence of prion infectivity by animal bioassay. None of the D. andersoni blood meals that had been ingested from prion-infected hamsters yielded evidence of prion infection. Overall, the data do not support a role of D. andersoni in the transmission of prion disease.

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Detection of prion protein in the cerebrospinal fluid of elk (Cervus canadensis nelsoni) with chronic wasting disease using protein misfolding cyclic amplification

Journal of Veterinary Diagnostic Investigation ◽

10.1177/1040638712448060 ◽

2012 ◽

Vol 24 (4) ◽

pp. 746-749 ◽

Cited By ~ 13

Author(s):

Tracy A. Nichols ◽

Terry R. Spraker ◽

Tom Gidlewski ◽

Jenny G. Powers ◽

Glenn C. Telling ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Prion Protein ◽

Protein Misfolding ◽

Chronic Wasting Disease ◽

Wasting Disease ◽

Protein Misfolding Cyclic Amplification ◽

Cervus Canadensis

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Experimental Transmission of the Chronic Wasting Disease Agent to Swine after Oral or Intracranial Inoculation

Journal of Virology ◽

10.1128/jvi.00926-17 ◽

2017 ◽

Vol 91 (19) ◽

Cited By ~ 14

Author(s):

S. Jo Moore ◽

M. Heather West Greenlee ◽

Naveen Kondru ◽

Sireesha Manne ◽

Jodi D. Smith ◽

...

Keyword(s):

Prion Protein ◽

Prion Disease ◽

Chronic Wasting Disease ◽

Mouse Bioassay ◽

Lymphoid Tissues ◽

Species Barrier ◽

Wasting Disease ◽

Disease Agent ◽

Domestic Swine

ABSTRACT Chronic wasting disease (CWD) is a naturally occurring, fatal neurodegenerative disease of cervids. The potential for swine to serve as hosts for the agent of CWD is unknown. The purpose of this study was to investigate the susceptibility of swine to the CWD agent following experimental oral or intracranial inoculation. Crossbred piglets were assigned to three groups, intracranially inoculated (n = 20), orally inoculated (n = 19), and noninoculated (n = 9). At approximately the age at which commercial pigs reach market weight, half of the pigs in each group were culled (“market weight” groups). The remaining pigs (“aged” groups) were allowed to incubate for up to 73 months postinoculation (mpi). Tissues collected at necropsy were examined for disease-associated prion protein (PrPSc) by Western blotting (WB), antigen capture enzyme immunoassay (EIA), immunohistochemistry (IHC), and in vitro real-time quaking-induced conversion (RT-QuIC). Brain samples from selected pigs were also bioassayed in mice expressing porcine prion protein. Four intracranially inoculated aged pigs and one orally inoculated aged pig were positive by EIA, IHC, and/or WB. By RT-QuIC, PrPSc was detected in lymphoid and/or brain tissue from one or more pigs in each inoculated group. The bioassay was positive in four out of five pigs assayed. This study demonstrates that pigs can support low-level amplification of CWD prions, although the species barrier to CWD infection is relatively high. However, detection of infectivity in orally inoculated pigs with a mouse bioassay raises the possibility that naturally exposed pigs could act as a reservoir of CWD infectivity. IMPORTANCE We challenged domestic swine with the chronic wasting disease agent by inoculation directly into the brain (intracranially) or by oral gavage (orally). Disease-associated prion protein (PrPSc) was detected in brain and lymphoid tissues from intracranially and orally inoculated pigs as early as 8 months of age (6 months postinoculation). Only one pig developed clinical neurologic signs suggestive of prion disease. The amount of PrPSc in the brains and lymphoid tissues of positive pigs was small, especially in orally inoculated pigs. Regardless, positive results obtained with orally inoculated pigs suggest that it may be possible for swine to serve as a reservoir for prion disease under natural conditions.

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