Proteolytic inactivation of the bovine spongiform encephalopathy agent

McLeod, A.; Murdoch, Heather; Dickinson, Joanne L.; Dennis, Mike; Hall, G.A.; Buswell, Clive M.; Carr, Jean K.; Taylor, David; Sutton, J. Mark; Raven, Neil

doi:10.1016/j.bbrc.2004.03.168

Cited by 68 publications

(67 citation statements)

References 25 publications

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“…McKinley et al [11] reported that proteinase K digestion for more than 2 h leads to hydrolysis. In recent publications, thermostable alkaline serine proteases of Thermococcus, Thermosipho, Thermoanaerobacter and Streptomyces were able to degrade PrP Sc under special conditions [7,12,21]. Denaturation leads to a conformation of PrP Sc susceptible to proteolysis.…”

Section: Discussionmentioning

confidence: 99%

“…PrP Sc is resistant towards other proteolytic enzymes like trypsin, V8 protease [11] and pepsin [8]. Recently, data were published indicating that extra cellular proteases of Streptomyces, Bacillus and anaerobic thermophilic prokaryotes are able to decrease the PrP Sc level in infected brain homogenates of mice and hamsters [7,8,11,12]. Thus, TSE agents are sensitive towards proteases, although such proteases rarely appear to occur.…”

Section: Introductionmentioning

confidence: 99%

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Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle

Scherbel¹,

Pichner²,

Groschup

et al. 2006

Vet. Res.

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-A food-borne origin of the transmission of bovine spongiform encephalopathy (BSE) to cattle is commonly assumed. However, the fate of infectious prion protein during polygastric digestion remains unclear. It is unknown at present, whether infectious prion proteins, considered to be very stable, are degraded or inactivated by microbial processes in the gastrointestinal tract of cattle. In this study, rumen and colon contents from healthy cattle, taken immediately after slaughter, were used to assess the ability of these microbial consortia to degrade PrP Sc . Therefore, the consortia were incubated with brain homogenates of scrapie (strain 263K) infected hamsters under physiological anaerobic conditions at 37• C. Within 20 h, PrP Sc was digested both with ruminal and colonic microbiota up to immunochemically undetectable levels. Especially polymyxin resistant (mainly gram-positive) bacteria expressed PrP Sc degrading activity. These data demonstrate the ability of bovine gastrointestinal microbiota to degrade PrP Sc during digestion.transmissible spongiform encephalopathy / prion / degradation / microbiota / gastrointestinal tract

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle

Scherbel¹,

Pichner²,

Groschup

et al. 2006

Vet. Res.

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“…Standard methods such as western blotting fail to detect significant levels of infectivity, [58][59][60] and the most sensitive method of prion detection, animal bioassay, would be impractical for use on large numbers of environmental samples. Protein misfolding cyclic amplification (PMCA), 61 developed by Soto and colleagues for detecting small amounts of PrP Sc , has generated much interest for use as an environmental detection method.…”

Section: Prion Diseases and The Prion Proteinmentioning

confidence: 99%

“…Bacterial enzymes which effectively degrade prions have been identified, but they are most effective at high pH (10-12) and high temperature (50-60°C). 59,104 Microbiological consortia taken from the rumen and colon of cattle could degrade PrP Sc to undetectable levels within 20 hours under anaerobic conditions at 37°C, although infectivity remained. 60,105 Using an enzyme treatment, it may be possible to lower or eliminate the infectivity at identified or presumed CWD and scrapie 'hot spots' in captive and wild settings.…”

Section: Degradation and Mitigation Of Prions In The Environmentmentioning

confidence: 99%

Prions in the environment

Saunders

Bartelt–Hunt

Bartz

2008

Prion

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Scrapie and CWD are horizontally transmissible, and the environment likely serves as a stable reservoir of infectious prions, facilitating a sustained incidence of CWD in free-ranging cervid populations and complicating efforts to eliminate disease in captive herds. Prions will enter the environment through mortalities and/ or shedding from live hosts. Unfortunately, a sensitive detection method to identify prion contamination in environmental samples has not yet been developed. An environmentally-relevant prion model must be used in experimental studies. Changes in PrP Sc structure upon environmental exposure may be as significant as changes in PrP Sc quantity, since the structure can directly affect infectivity and disease pathology. Prions strongly bind to soil and remain infectious. Conformational changes upon adsorption, competitive sorption and potential for desorption and transport all warrant further investigation. Mitigation of contaminated carcasses or soil might be accomplished with enzyme treatments or composting in lieu of incineration.

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“…Protease digestion and elimination of ELISA or Western-blot immunoreactivity has been shown under a variety of conditions (Hui et al, 2004a(Hui et al, , 2004bLangeveld et al, 2003;Tsiroulnikov et al, 2004;Mitsuiki et al, 2006;Rapp et al, 2006;Scherbel et al, 2006;Muller-Hellwig et al, 2006). However, protease destruction of prions below in vitro assay detection limits does not necessarily mean that PrP res has been reduced to levels that are no longer infectious in more sensitive bioassays; therefore, methods investigating decontamination of infectious prions need to be tested in vivo until better in vitro methods are developed and validated (McLeod et al, 2004;Lawson et al, 2006). Bioassays in live animals are more sensitive, test actual infection, are representative of natural infection, and can detect low-level residual infectivity that may reside below the limit of detection of traditional immunoassays.…”

Section: Introductionmentioning

confidence: 99%

Feasibility of infectious prion digestion using mild conditions and commercial subtilisin

Pilon

Nash²,

Arver³

et al. 2009

Journal of Virological Methods

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a b s t r a c tTwo serine protease enzymes, subtilisin 309 and subtilisin 309-v, were used to digest brain homogenates containing high levels of prion infectivity using mildly alkaline conditions to investigate prion decontamination methods. To establish that PrP res infectivity was eliminated, we utilized the Rocky Mountain Laboratory (RML) mouse-adapted scrapie model system for bioassay. Only one digestion condition (subtilisin 309 at 138 mAU/ml, 55• C and 14 h digestion time pH 7.9) was considered to be highly relevant statistically (P < 0.001) compared to control, with 52% of challenged mice surviving until the end of the study period. In contrast, treatment of PrP res by autoclaving at 134• C or treatment with hypochlorite at a concentration of 20,000 ppm completely protected mice from prionosis. Further, in vitro assays suggest that potential proteolytic based PrP res decontamination methods must use high enzyme concentration, pH values >9.0, and elevated temperatures to be a safely efficacious, thereby limiting applicability on delicate surgical instruments and use in the environment.Published by Elsevier B.V.

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Proteolytic inactivation of the bovine spongiform encephalopathy agent

Cited by 68 publications

References 25 publications

Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle

Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle

Prions in the environment

Feasibility of infectious prion digestion using mild conditions and commercial subtilisin

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Proteolytic inactivation of the bovine spongiform encephalopathy agent

Cited by 68 publications

References 25 publications

Degradation of scrapie associated prion protein (PrPSc) by the gastrointestinal microbiota of cattle

Degradation of scrapie associated prion protein (PrPSc) by the gastrointestinal microbiota of cattle

Prions in the environment

Feasibility of infectious prion digestion using mild conditions and commercial subtilisin

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Product

Resources

About

Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle

Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle