Classical Scrapie Did Not Re-occur in Goats After Cleaning and Disinfection of the Farm Premises

After an outbreak of classical scrapie in a dairy goat herd with over 1,800 goats, all goats in the herd were culled in 2008, cleaning and disinfection of the premises was implemented, and restocking with goats took place ~4 months after depopulation. Ten years later the new herd population is over 3,000 goats. This study was carried out to determine whether the measures were effective to prevent re-occurrence of scrapie to the 1% prevalence level seen when scrapie was first detected on this farm. A total of 280 goats with a minimum age of 18 months, which were predominantly at the end of their productive life, were euthanized, and brain and retropharyngeal lymph node examined by immunohistochemistry for disease-associated prion protein. Genotyping was done in all euthanized goats and live male goats used or intended for breeding to determine prion protein gene polymorphisms associated with resistance to classical scrapie. None of the goats presented with disease-associated prion protein in the examined tissues, and 34 (12.2%) carried the K222 allele associated with resistance. This allele was also found in four breeding male goats. The study results suggested that classical scrapie was not re-introduced on this goat farm through mass restocking or inadequate cleaning and disinfection procedures. Further scrapie surveillance of goats on this farm is desirable to confirm absence of disease. Breeding with male goats carrying the K222 allele should be encouraged to increase the scrapie-resistant population

Estimating the impact on food and edible materials of changing scrapie control measures : the scrapie control model

Multiple controls established during the bovine spongiform encephalopathy (BSE) epidemic were not solely applied to BSE in cattle, but were implemented for scrapie in sheep and goats due to concerns over the occurrence of BSE in sheep. In the absence of BSE in sheep being observed, control measures for prion diseases are now being evaluated to ensure they remain proportionate to risk. This risk assessment, aims to estimate, by use of stochastic simulation, the impact of reducing controls for Specified Risk Materials (SRM) from sheep at abattoir. Three scenarios have been included: 1) current list of SRM; 2) brain and spinal cord of adult sheep; and 3) the brain of adult sheep. Results indicate the total amount of infectivity passing through British abattoirs is highest for atypical scrapie with nearly 3,500,000 Ovine Oral (OO) ID50 per year. The majority of this infectivity enters Category 1 waste for incineration, with only 13,000 OO ID50 per year within edible products. Under Scenario 2, an additional 4,000 OO ID50 per year would be classified as edible products from the lifting of restrictions on the distal ileum of adult sheep. However, if SRM removal was limited to brain, an additional 110,000 OO ID50 per year would be permitted into edible products with the lifting of restrictions on the spinal cord of adult sheep. For classical scrapie, there is a mean estimate of infectivity of 30,000 OO ID50 per year at abattoir. This is lower than for atypical scrapie due to the lower occurrence of this disease in Britain. However, more infectivity is destined to reach the food chain as the disease is peripherally distributed in the carcase. The highest contributor to the total amount of infectivity consumed per year is the intestines (duodenum and jejunum). If SRM removal is limited to the brain and spinal cord of sheep over 12 months of age, there is an approximate mean increase from 19,000 to 21,000 OO ID50 per year diverted to edible products. If the SRM list is restricted to brain only, this increases to over 23,000 OO ID50 per year. For the potential of sheep-BSE, there is a very low estimate of 29 OO ID50 per year in total from carcases entering abattoir, due to the potential very rare occurrence of this disease. Given changes in SRM regulations there is a change of an additional 4 OO ID50 per year being diverted to edible products

A review of cleaning and disinfection guidelines and recommendations following an outbreak of classical scrapie

[EN] Classical scrapie is a prion disease of small ruminants, the infectious agent of which has been shown to be extremely persistent in the environment. Cleaning and disinfection (C&D) after a scrapie outbreak is currently recommended by many governments' veterinary advisors and implemented in most farms affected. Yet, the effectiveness of these procedures remains unclear. The aim of this study was to review existing literature and guidelines regarding farm C&D protocols following classical scrapie outbreaks and assess their effectiveness and the challenges that translation of policy and legislative requirements present at a practical level. A review of the literature was conducted to identify the on-farm C&D protocols used following outbreaks of scrapie, assess those materials with high risk for persistence of the scrapie agent on farms, and review the existing evidence of the effectiveness of recommended C&D protocols. An expert workshop was also organised in Great Britain (GB) to assess: the decision-making process used when implementing C&D protocols on GB farms, the experts' perceptions on the effectiveness of these protocols and changes needed, and their views on potential recommendations for policy and research. Outputs of the literature review revealed that the current recommended protocol for C&D [1 h treatment with sodium hypochlorite containing 20,000 ppm free chlorine or 2 M sodium hydroxide (NaOH)] is based on labo-ratory experiments. Only four field farm experiments have been conducted, indicating a lack of data on effec-tiveness of C&D protocols on farms by the re-occurrence of scrapie infection post re-stocking. Recommendations related to the control of outdoor environment, which are difficult and expensive to implement, vary between countries. The expert workshop concluded that there are no practical, cost-effective C&D alternatives to be considered at this time, with control therefore based on C&D only in combination with additional time restrictions on re-stocking and replacement with non-susceptible livestock or more genetically resistant types, where available. Participants agreed that C&D should still be completed on scrapie affected farms, as it is considered to be "good disease practice" and likely to reduce the levels of the prion protein. Participants felt that any additional protocols developed should not be "too prescriptive" (should not be written down in specific policies) because of significant variation in farm types, farm equipment and installations. Under this scenario, control of classical scrapie on farms should be designed with a level of C&D in combination with re-stocking temporal ban and replacement with livestock of limited susceptibility.The APHA would like to thank all the expert contributions at the workshop. This study was funded by Defra (UK) under Project SE1960 and the Government of education and science of the Generalitat Valenciana (Spain)Alarcon, P.; Marco-Jiménez, F.; Horigan, V.; Ortiz-Pelaez, A.; Rajanayagam, B.; Dryden, A.; Simmons, H.... (2021). A review of cleaning and disinfection guidelines and recommendations following an outbreak of classical scrapie. Preventive Veterinary Medicine. 193:1-9. https://doi.org/10.1016/j.prevetmed.2021.1053881919

The Potential of Biomarkers as Indices of Response to Naturally Occurring Toxicants (Including Those Produced by Cooking).

The potential of biomarkers to measure overall responses to dietary/environmental toxicants and putative protective factors has been investigated in vitro and in vivo. In stability studies, isolated lymphocytes in phosphate-buffered saline were stable overnight at 4°C if protected from light and stained lymphocytes were stable up to 120 hours in the lysing solution. In the Comet assay, human lymphocytes in vitro showed a concentration-related response to hydrogen peroxide or N-methyl-nitro-N'-nitrosoguanidine. Caffeine did not induce DNA damage even after activation by Aroclor 1254-induced S9 mix but the solvents used to remove it from green tea (chloroform and dichloromethane) did so even without metabolic activation. Dietary carcinogens 2-amino-3-methyl-[4,5-f]quinoline (IQ), Ochratoxin A (OTA) and mushroom and its metabolites were tested for genotoxicity in vitro with human lymphocytes in the Comet assay. A concentration-related response was associated with IQ, OTA, and raw mushroom extract. Baked mushroom extracts were effective only at high concentrations but the effects of agaritine did not reach statistical significance. 4-(Hydroxymethyl)benzenediazonium ion produced considerably more DNA damage than equimolar amounts of agaritine or N'-acetyl-4-(hydroxymethyl)phenyl-hydrazine. Green tea extract in vitro inhibited IQ-induced DNA damage in human lymphocytes using the Comet assay. Green tea flavonoids showed different efficacy against IQ genotoxicity. Caffeine-induced S9 mix and Aroclor 1254-induced S9 mix activated IQ to a similar extent. The DNA synthesis inhibitor aphidicolin failed to increase the sensitivity of the Comet assay. A preliminary in vivo study using male Wistar albino rats showed modulatory effects of green tea extract against IQ (1 mg/kg b.wt) in the Comet assay and TBA test. On examination of the colon no aberrant crypt foci were observed. HPLC profiling showed specific peaks for green tea and IQ metabolites however, the IQ specific peaks were not identified in the IQ+green tea (GT) group. In a more comprehensive study using a higher level of IQ (5 mg/kg b.wt) similar modulatory effects of green tea were observed in the Comet assay and TBA test. The TBA test using HPLC and colorimetric methods demonstrated that although the trends were similar the values were higher with the latter method. HPLC profiling showed chromatographic patterns similar to the trial study however the enzyme digestion identified conjugates especially sulphate conjugates with IQ and IQ+GT group. This in vivo study demonstrates that green tea (2.5% w/v) is capable of reducing IQ-induced genotoxicity and lipid peroxidation when given simultaneously