Transmission Of Scrapie Research Articles

Sparse PrPSc accumulation in the placentas of goats with naturally acquired scrapie

BackgroundDomestic goats (Capra hircus) are a natural and experimental host of scrapie and bovine spongiform encephalopathy, the transmissible spongiform encephalopathies (TSE) of sheep and cattle. Goats are also susceptible to experimental infection with the agents of TSEs of deer and elk (chronic wasting disease) and humans (Creutzfeldt Jakob disease). Distribution of PrPSc, the abnormal prion protein, is similar in the tissues of scrapie-infected sheep and goats but no data are available on the potential shedding of the agent through the placenta, the presumed route of transmission of ovine scrapie. We describe the sparse accumulation of PrPSc in the placentas of goats with naturally acquired classical scrapie in comparison to field cases of classical ovine scrapie.ResultsPrPSc was detected in the shed placentas from a sample of U.S. goats with naturally occurring scrapie, diagnosed by antemortem lymphoid tissue biopsy or identified as high risk progeny of infected dams. PrPSc accumulation patterns in the intact placentome and western blot banding was similar in the caprine and ovine samples. However, levels of PrPSc estimated from ELISA and immunohistochemistry assays were generally lower in goats than in sheep, although wide variation was noted in both species.ConclusionsPrPSc accumulates in the shed placentas of goats with naturally acquired scrapie. Although these levels were low in most caprine samples, the caprine placenta may contribute to prion contamination of kidding facilities and transmission to co-housed sheep or goats.

Breeding with resistant rams leads to rapid control of classical scrapie in affected sheep flocks

Susceptibility to scrapie, a transmissible spongiform encephalopathy in sheep, is modulated by the genetic make-up of the sheep. Scrapie control policies, based on selecting animals of resistant genotype for breeding, have recently been adopted by the Netherlands and other European countries. Here we assess the effectiveness of a breeding programme based on selecting rams of resistant genotype to obtain outbreak control in classical scrapie-affected sheep flocks under field conditions. In six commercially-run flocks following this breeding strategy, we used genotyping to monitor the genotype distribution, and tonsil biopsies and post-mortem analyses to monitor the occurrence of scrapie infection. The farmers were not informed about the monitoring results until the end of the study period of six years. We used a mathematical model of scrapie transmission to analyze the monitoring data and found that where the breeding scheme was consistently applied, outbreak control was obtained after at most four years. Our results also show that classical scrapie control can be obtained before the frequency of non-resistant animals is reduced to zero in the flock. This suggests that control at the national scale can be obtained without a loss of genetic polymorphisms from any of the sheep breeds.

The Interaction of Ruminant PrPSc with Soils Is Influenced by Prion Source and Soil Type

The persistence of prions within the environment is implicated in the horizontal transmission of ovine scrapie and cervid chronic wasting disease. Description of the interaction of prion strains derived from their natural hosts with a range of soil types is imperative in understanding how prions persist in the environment and, therefore, the characteristics of prion transmission. Here, we demonstrate that all detectable ovine scrapie and bovine BSE PrP(Sc) bind to a range of soil types within 24 h. This highly efficient binding of prions to soils is characterized by truncation of desorbed PrP(Sc) in a soil-dependent manner, with clay-rich soils resulting in N-terminal truncation of the PrP(Sc) and sand-rich soils yielding full length PrP(Sc) species. PrP(Sc) did not migrate through soil columns during incubation for up to 18 months, and for all combinations of soil and prion types, a decrease in recoverable PrP(Sc) was seen over time. Persistence of PrP(Sc) within soil and their interaction with soil particles of distinct sizes was dictated by both the soil type and the source of the prion, with ovine scrapie being apparently more persistent in some soils than cattle BSE. These data indicate that natural ruminant prion strains are stable in the soil environment for at least 18 months and that PrP(Sc)-soil interaction is dictated by both the soil properties and the strain/host species of PrP(Sc).

A “shotgun” method for tracing the birth locations of sheep from flock tags, applied to scrapie surveillance in Great Britain

Movement records are often used to identify animal sample provenance by retracing the movements of individuals. Here we present an alternative method, which uses the same identity tags and movement records as are used to retrace movements, but ignores individual movement paths. The first step uses a simple query to identify the most likely birth holding for every identity tag included in a database recording departures from agricultural holdings. The second step rejects a proportion of the birth holding locations to leave a list of birth holding locations that are relatively reliable. The method was used to trace the birth locations of sheep sampled for scrapie in abattoirs, or on farm as fallen stock. Over 82% of the sheep sampled in the fallen stock survey died at the holding of birth. This lack of movement may be an important constraint on scrapie transmission. These static sheep provided relatively reliable birth locations, which were used to define criteria for selecting reliable traces. The criteria rejected 16.8% of fallen stock traces and 11.9% of abattoir survey traces. Two tests provided estimates that selection reduced error in fallen stock traces from 11.3% to 3.2%, and in abattoir survey traces from 8.1% to 1.8%. This method generated 14,591 accepted traces of fallen stock from samples taken during 2002–2005 and 83,136 accepted traces from abattoir samples. The absence or ambiguity of flock tag records at the time of slaughter prevented the tracing of 16–24% of abattoir samples during 2002–2004, although flock tag records improved in 2005. The use of internal scoring to generate and evaluate results from the database query, and the confirmation of results by comparison with other database fields, are analogous to methods used in web search engines. Such methods may have wide application in tracing samples and in adding value to biological datasets.

Transport of the Pathogenic Prion Protein through Soils

Transmissible spongiform encephalopathies (TSEs) are progressive neurodegenerative diseases and include bovine spongiform encephalopathy of cattle, chronic wasting disease (CWD) of deer and elk, scrapie in sheep and goats, and Creutzfeldt-Jakob disease in humans. An abnormally folded form of the prion protein (designated PrP(TSE)) is typically associated with TSE infectivity and may constitute the major, if not sole, component of the infectious agent. Transmission of CWD and scrapie is mediated in part by an environmental reservoir of infectivity. Soil appears to be a plausible candidate for this reservoir. The transport of TSE agent through soil is expected to influence the accessibility of the pathogen to animals after deposition and must be understood to assess the risks associated with burial of infected carcasses. We report the results of saturated column experiments designed to evaluate PrP(TSE) transport through five soils with relatively high sand or silt contents and low organic carbon content. Protease-treated TSE-infected brain homogenate was used as a model for PrP(TSE) present in decomposing infected tissue. Synthetic rainwater was used as the eluent. All five soils retained PrP(TSE); no detectable PrP(TSE) was eluted over more than 40 pore volumes of flow. Lower bound apparent attachment coefficients were estimated for each soil. Our results suggest that TSE agent released from decomposing tissues to soils with low organic carbon content would remain near the site of initial deposition. In the case of infected carcasses deposited on the land surface, this may result in local sources of infectivity to other animals.

Use of a preclinical test in the control of classical scrapie

Scrapie control in Great Britain (GB) was originally based on the National Scrapie Plan's Ram Genotyping scheme aimed at reducing the susceptibility of the national flock. The current official strategy to control scrapie in the national flock involves culling susceptible genotypes in individual, known affected flocks (compulsory scrapie flock scheme or CSFS). However, the recent development of preclinical test candidates means that a strategy based on disease detection may now be feasible. Here, a deterministic within-flock model was used to demonstrate that only large flocks with many home-bred ewes are likely to be a significant risk for flock-to-flock transmission of scrapie. For most other flocks, it was found that the CSFS could be replaced by a strategy using a currently available live test without excessive risk to other farmers, even if the proportion of susceptible genotypes in the flock is unusually large. Even for flocks that represent a high risk of harbouring a high prevalence of infection, there would be limited probability of onward transmission if scrapie is detected soon after disease introduction (typically less than 5 years). However, if detection of disease is delayed, the existing CSFS strategy may be the most appropriate control measure in these cases.

Prions Are Secreted into the Oral Cavity in Sheep with Preclinical Scrapie

A major concern in prion disease transmission is the spread of the disease agent by means of secretions and excretions. We analyzed buccal swab samples obtained from preclinical scrapie-infected sheep by concentrating the collected prions on silicon dioxide, followed by amplification by serial protein misfolding cyclic amplification. Data clearly demonstrate that prions are present in buccal swab samples from sheep with a VRQ/VRQ PRNP genotype during preclinical scrapie infection. These data describe for the first time to our knowledge the secretion of prions into the oral cavity of sheep, a finding with implications for the transmission of ovine scrapie and very likely other prion diseases.

Ovine PrP Genotype is Linked With Lesion Profile and Immunohistochemistry Patterns After Primary Transmission of Classical Scrapie to Wild-Type Mice

It is currently believed that primary transmission of classical scrapie to wild-type mice is inefficient and characterized by low attack rates and variable incubation periods and lesion profiles. Consequently, strain characterization of classical scrapie in these mice relies on subpassage. The aim of this study was to perform a retrospective analysis of lesion profiles and immunohistochemistry patterns after transmission of a large number of classical scrapie sources to wild-type mice and to investigate trends that might be used to characterize the agent without subpassaging. Scrapie field cases (n = 31) collected from individual farms between 1996 and 1999 were inoculated into RIII, C57BL, and VM mice and profiled using standard methodology and analyzed by immunohistochemistry. Using cluster analysis to resultant lesion profiles produced groups of similar lesion profiles in RIII and C57BL mice. We observed correlations between lesion profile clusters and the ovine prion protein (PrP) genotype. Immunohistochemistry indicated donor-mediated trends in the PrP pattern. These results indicate that ovine PrP genotype is a factor that is linked to both the lesion profile and the pattern of PrP deposition on primary transmission of classical scrapie to wild-type mice.

Pathogenesis of natural goat scrapie: modulation by hostPRNPgenotype and effect of co-existent conditions

After detection of a high prevalence of scrapie in a large dairy goat herd, 72 infected animals were examined by immunohistochemistry with prion protein (PrP) antibody Bar224 to study the pathogenesis of the infection. Tissues examined included the brain and thoracic spinal cord (TSC), a wide selection of lymphoreticular system (LRS) tissues, the distal ileum and its enteric nervous system (ENS), and other organs, including the mammary gland. The whole open reading frame of the PRNP gene was sequenced and antibodies to caprine arthritis-encephalitis virus (CAEV) infection were determined. Unexpectedly, accumulation of disease-associated PrP (PrPd) in the brain was more frequent in methionine carriers at codon 142 (24/32, 75.0%) than amongst isoleucine homozygotes (14/40, 35.0%). The latter, however, showed significantly greater amounts of brain PrPd than the former (average scores of 9.3 and 3.0, respectively). A significant proportion of the 38 goats that were positive in brain were negative in the ENS (44.7%) or in the TSC (39.5%). These results, together with the early and consistent involvement of the circumventricular organs and the hypothalamus, point towards a significant contribution of the haematogenous route in the process of neuroinvasion. Chronic enteritis was observed in 98 of the 200 goats examined, with no association with either scrapie infection or presence of PrPd in the gut. Lymphoproliferative interstitial mastitis was observed in 13/31 CAEV-positive and scrapie-infected goats; PrPd in the mammary gland was detected in five of those 13 goats, suggesting a possible contribution of CAEV infection in scrapie transmission via milk.

Eradication of scrapie with selective breeding: are we nearly there?

BackgroundFollowing EU decision 2003/100/EC Member States have recently implemented sheep breeding programmes to reduce the prevalence of sheep with TSE susceptible prion genotypes. The present paper investigates the progress of the breeding programme in the Netherlands. The PrP genotype frequencies were monitored through time using two sets of random samples: one set covers the years 2005 to 2008 and is taken from national surveillance programme; the other is taken from 168 random sheep farms in 2007. The data reveal that although the level of compliance to the breeding programme has been high, the frequency of susceptible genotypes varies substantially between farms. The 168 sheep farms are a subset of 689 farms participating in a postal survey inquiring about management and breeding strategies. This survey aimed to identify how much these strategies varied between farms, in order to inform assessment of the expected future progress towards eradication of classical scrapie.ResultsOn the one hand, we found that compliance to the national breeding program has been high, and the frequency of resistant genotypes is expected to increase further in the next few years. On the other hand, we observed a large variation in prevalence of the scrapie resistant PrP genotype ARR between farms, implicating a large variation of genetic resistance between farms. Substantial between-flock differences in management and breeding strategies were found in the postal survey, suggesting considerable variation in risk of scrapie transmission between farms.ConclusionsOur results show that although there has been a good progress in the breeding for scrapie resistance and the average farm-level scrapie susceptibility in the Netherlands has been significantly reduced, still a considerable proportion of farms contain high frequencies of susceptible genotypes in their sheep population. Since 2007 the breeding for genetic resistance is voluntarily again, and participation to selective breeding can decrease as a result of this. This, together with the patterns of direct and indirect contact between sheep farms, might present a challenge of the aim of scrapie eradication. Communication to sheep owners of the effect of the breeding programme thus far, and of the prospects for classical scrapie eradication in The Netherlands might be essential for obtaining useful levels of participation to the voluntary continuation of the breeding programme.

Evidence for maternal transmission of scrapie in naturally affected flocks

It has been known for many years that the offspring of scrapie affected ewes are at increased risk of developing scrapie but whether this is simply the result of an increased genetic susceptibility or transmission of infection has always been unclear. To contribute to clarify this we analysed the data collected in a detailed study of scrapie occurrence in a number of naturally affected commercial sheep flocks in Great Britain (GB) to investigate the association between PrP genotype and parental scrapie status and the incidence of scrapie. Our analyses confirmed the strong association between PrP genotype and the incidence of scrapie found in previous studies and a low incidence of scrapie in animals carrying the ARR allele and a high risk in homozygous VRQ animals. However, we also demonstrate an increased incidence of scrapie in the offspring of scrapie affected ewes controlling for the confounding effect of PrP genotype, but no increased scrapie incidence in the offspring of scrapie affected sires. Our results suggest that some of the increased incidence of scrapie in the offspring of scrapie affected ewes is the result of transmission of infection from mother to offspring. Our results confirm that a breeding policy aimed at decreasing the genetic susceptibility of the population should decrease the incidence of scrapie and that removing the offspring of scrapie affected animals from affected flocks could contribute to the control of this disease.

The evaluation of exposure risks for natural transmission of scrapie within an infected flock

BackgroundAlthough the epidemiology of scrapie has been broadly understood for many years, attempts to introduce voluntary or compulsory controls to eradicate the disease have frequently failed. Lack of precision in defining the risk factors on farm has been one of the challenges to designing control strategies. This study attempted to define which parts of the annual flock management cycle represented the greatest risk of infection to naive lambs exposed to the farm environment at different times.ResultsIn VRQ/VRQ lambs exposed to infected sheep at pasture or during lambing, and exposed to the buildings in which lambing took place, the attack rate was high and survival times were short. Where exposure was to pasture alone the number of sheep affected in each experimental group was reduced, and survival times were longer and related to length of exposure.ConclusionAt the flock level, eradication and control strategies for scrapie must take into account the need to decontaminate buildings used for lambing, and to reduce (or prevent) the exposure of lambs to infected sheep, especially in the later stages of incubation, and at lambing. The potential for environmental contamination from pasture should also be considered. Genotype selection may still prove to be the only viable tool to prevent infection from contaminated pasture, reduce environmental contamination and limit direct transmission from sheep to sheep.

Transmissibility of Atypical Scrapie in Ovine Transgenic Mice: Major Effects of Host Prion Protein Expression and Donor Prion Genotype

Atypical scrapie or Nor98 has been identified as a transmissible spongiform encephalopathy (TSE) that is clearly distinguishable from classical scrapie and BSE, notably regarding the biochemical features of the protease-resistant prion protein PrPres and the genetic factors involved in susceptibility to the disease. In this study we transmitted the disease from a series of 12 French atypical scrapie isolates in a transgenic mouse model (TgOvPrP4) overexpressing in the brain ∼0.25, 1.5 or 6× the levels of the PrPARQ ovine prion protein under the control of the neuron-specific enolase promoter. We used an approach based on serum PrPc measurements that appeared to reflect the different PrPc expression levels in the central nervous system. We found that transmission of atypical scrapie, much more than in classical scrapie or BSE, was strongly influenced by the PrPc expression levels of TgOvPrP4 inoculated mice. Whereas TgOvPrP4 mice overexpressing ∼6× the normal PrPc level died after a survival periods of 400 days, those with ∼1.5× the normal PrPc level died at around 700 days. The transmission of atypical scrapie in TgOvPrP4 mouse line was also strongly influenced by the prnp genotypes of the animal source of atypical scrapie. Isolates carrying the AF141RQ or AHQ alleles, associated with increased disease susceptibility in the natural host, showed a higher transmissibility in TgOvPrP4 mice. The biochemical analysis of PrPres in TgOvPrP4 mouse brains showed a fully conserved pattern, compared to that in the natural host, with three distinct PrPres products. Our results throw light on the transmission features of atypical scrapie and suggest that the risk of transmission is intrinsically lower than that of classical scrapie or BSE, especially in relation to the expression level of the prion protein.

Evaluation of the possible transmission of BSE and scrapie to gilthead sea bream (Sparus aurata).

In transmissible spongiform encephalopathies (TSEs), a group of fatal neurodegenerative disorders affecting many species, the key event in disease pathogenesis is the accumulation of an abnormal conformational isoform (PrPSc) of the host-encoded cellular prion protein (PrPC). While the precise mechanism of the PrPC to PrPSc conversion is not understood, it is clear that host PrPC expression is a prerequisite for effective infectious prion propagation. Although there have been many studies on TSEs in mammalian species, little is known about TSE pathogenesis in fish. Here we show that while gilthead sea bream (Sparus aurata) orally challenged with brain homogenates prepared either from a BSE infected cow or from scrapie infected sheep developed no clinical prion disease, the brains of TSE-fed fish sampled two years after challenge did show signs of neurodegeneration and accumulation of deposits that reacted positively with antibodies raised against sea bream PrP. The control groups, fed with brains from uninfected animals, showed no such signs. Remarkably, the deposits developed much more rapidly and extensively in fish inoculated with BSE-infected material than in the ones challenged with the scrapie-infected brain homogenate, with numerous deposits being proteinase K-resistant. These plaque-like aggregates exhibited congophilia and birefringence in polarized light, consistent with an amyloid-like component. The neurodegeneration and abnormal deposition in the brains of fish challenged with prion, especially BSE, raises concerns about the potential risk to public health. As fish aquaculture is an economically important industry providing high protein nutrition for humans and other mammalian species, the prospect of farmed fish being contaminated with infectious mammalian PrPSc, or of a prion disease developing in farmed fish is alarming and requires further evaluation.

Failure to transmit scrapie infection by transferring preimplantation embryos from naturally infected donor sheep

The objective of the study was to examine whether or not the preimplantation embryo can act as a carrier of classic scrapie infection. The study was carried out on quarantined premises with sheep of highly susceptible scrapie genotypes. Uninfected embryos, collected from New Zealand–derived Suffolk ewes, were surgically transferred into recipient ewes that were also of New Zealand origin. Seventeen negative control lambs were born on the study premises from these embryo transfers. Thirty-nine experimental lambs were from embryos collected from naturally infected donor ewes. The experimental lambs were also born on the study premises after their surgical transfer into recipient ewes of New Zealand origin. These embryos had been collected from donor ewes in a scrapie-infected flock where the ewes were clinically sick with scrapie or developed clinical scrapie after embryo collection. All lambs were confirmed as scrapie susceptible of the ARQ/ARQ genotype. Twenty-eight experimental animals survived to the end point of the study at 5 yr of age with a mean survival of 1579 d. In the negative control group, 12 of 17 sheep survived to 5 yr of age with a mean survival of 1508 d. Postmortem examinations were carried out on all animals derived by embryo transfer, and in none was histologic or immunohistochemical evidence of scrapie found. In contrast, in the originating flock the majority of scrapie cases occurred in ARQ/ARQ genotyped animals where a 56% mortality from scrapie had been recorded in animals of this genotype. Thus, the study provides no evidence for transmission of scrapie and reinforces published evidence that vertical transmission of scrapie may be circumvented by embryo transfer procedures.

Hay Mite Communities in Sheep Farms from French and Spanish Basque Regions with regard to Scrapie Infection

It has been suggested that the mites connected with the decay of stored hay may be involved in the transmission of scrapie, a neurological disorder in sheep. To find out whether there are any obvious differences in the mite fauna in hay that could be related to outbreaks of scrapie in sheep, we analysed the hay fauna from 23 farms in the Basque regions in France and Spain, areas with high and very low incidence of scrapie, respectively. We did find some differences in the composition of the fauna, and a species such as Pygmephorus spp. was more prevalent in scrapie-infected farms, and particularly in highly scrapie-infected farms. The former species is associated with late stages in the biological degradation of stored hay and could be linked to the disease. Besides being the first record of the composition of the hay mite fauna in Spain and France our finding of differences in mite communities in hay from scrapie versus non-scrapie farms is a further indication of the possible association of certain mite species with scrapie.

Transmission of scrapie and sheep-passaged bovine spongiform encephalopathy prions to transgenic mice expressing elk prion protein

Chronic wasting disease (CWD) is a transmissible, fatal prion disease of cervids and is largely confined to North America. The origin of CWD continues to pose a conundrum: does the disease arise spontaneously or result from some other naturally occurring reservoir? To address whether prions from sheep might be able to cause disease in cervids, we inoculated mice expressing the elk prion protein (PrP) transgene [Tg(ElkPrP) mice] with two scrapie prion isolates. The SSBP/1 scrapie isolate transmitted disease to Tg(ElkPrP) mice with a median incubation time of 270 days, but a second isolate failed to produce neurological dysfunction in these mice. Although prions from cattle with bovine spongiform encephalopathy (BSE) did not transmit to the Tg(ElkPrP) mice, they did transmit after being passaged through sheep. In Tg(ElkPrP) mice, the sheep-passaged BSE prions exhibited an incubation time of approximately 300 days. SSBP/1 prions produced abundant deposits of the disease-causing PrP isoform, denoted PrP(Sc), in the cerebellum and pons of Tg(ElkPrP) mice, whereas PrP(Sc) accumulation in Tg mice inoculated with sheep-passaged BSE prions was confined to the deep cerebellar nuclei, habenula and the brainstem. The susceptibility of 'cervidized' mice to 'ovinized' prions raises the question about why CWD has not been reported in other parts of the world where cervids and scrapie-infected sheep coexist.

Frequency and Distribution of Nerves in Scrapie-affected and Unaffected Peyer's Patches and Lymph Nodes

Transmission of sheep scrapie and some other prion diseases, including variant Creutzfeldt-Jakob disease of man, probably occurs via the oral route. A disease-associated variant of the host-coded prion protein (PrP(d)) accumulates in germinal center follicles of lymphoid tissues, including Peyer's patches of the gut, where it can be detected before its accumulation in the central nervous system. To investigate the potential role of lymphoid tissue nerves in neuroinvasion, we used immunohistochemical methods to study the frequency and distribution of nerves and PrP(d) accumulation in Peyer's patches and other lymphoid tissues from scrapie-affected and unaffected sheep. Nerves were infrequently found in secondary follicles of Peyer's patches, but never in germinal centers of the other lymphoid tissues tested. No differences in the frequency or distribution of nerves were found in relation to the presence or absence of PrP(d) accumulation. PrP(d) accumulation and nerves were only infrequently present together in Peyer's patches. These results suggest that, even if amplification of infectivity in lymphoid tissues facilitates neuroinvasion, nerves within lymph nodes and germinal centers of Peyer's patches do not play a primary role in transport of infectivity to the central nervous system. However, sheep between 2 and 4 months of age had significantly more nerve fibers within follicles than older groups. It is therefore possible that a general increase in nerve density of the intestine during early phases of life may contribute to an increased susceptibility of young animals to oral prion infection.

Frequency and Distribution of Nerves in Scrapie-affected and Unaffected Peyer's Patches and Lymph Nodes

Transmission of sheep scrapie and some other prion diseases, including variant Creutzfeldt-Jakob disease of man, probably occurs via the oral route. A disease-associated variant of the host-coded prion protein (PrPd) accumulates in germinal center follicles of lymphoid tissues, including Peyer's patches of the gut, where it can be detected before its accumulation in the central nervous system. To investigate the potential role of lymphoid tissue nerves in neuroinvasion, we used immunohistochemical methods to study the frequency and distribution of nerves and PrPd accumulation in Peyer's patches and other lymphoid tissues from scrapie-affected and unaffected sheep. Nerves were infrequently found in secondary follicles of Peyer's patches, but never in germinal centers of the other lymphoid tissues tested. No differences in the frequency or distribution of nerves were found in relation to the presence or absence of PrPd accumulation. PrPd accumulation and nerves were only infrequently present together in Peyer's patches. These results suggest that, even if amplification of infectivity in lymphoid tissues facilitates neuroinvasion, nerves within lymph nodes and germinal centers of Peyer's patches do not play a primary role in transport of infectivity to the central nervous system. However, sheep between 2 and 4 months of age had significantly more nerve fibers within follicles than older groups. It is therefore possible that a general increase in nerve density of the intestine during early phases of life may contribute to an increased susceptibility of young animals to oral prion infection.

Experimental Oral Transmission of United States Origin Scrapie to Neonatal Sheep

Scrapie, a transmissible spongiform encephalopathy (TSE), is a naturally occurring fatal neurodegenerative disease of sheep and goats. The current study documents incubation periods, pathologic findings, and distribution of abnormal prion proteins (PrP(Sc)) by immunohistochemistry and Western blot in tissues of genetically susceptible and resistant neonatal lambs inoculated with pooled brain homogenates from 13 U.S. origin scrapie-affected ewes. Nine Suffolk lambs with genotypes AA/RR/QQ (n = 5) and AA/RR/QR (n = 4) at codons 136, 154, and 171, respectively) were orally inoculated, within 12 hr of birth, with 1 ml of a 10% (w/v) brain homogenate prepared from scrapie-affected sheep brains. Inoculated animals were euthanized when advanced clinical signs of scrapie were observed. All QQ sheep developed clinical signs of scrapie, with a mean survival time of 24 months. Spongiform lesions in the brains and PrP(Sc) deposits in the central nervous system and lymphoid tissues were present in these sheep. None of the QR sheep succumbed to the disease. A previous study that used a larger volume (30 ml of 10% brain suspension) of the same inoculum in 4-month-old Suffolk lambs of susceptible genotype documented longer survival periods (average 32 months), and only 5 of 9 inoculated sheep developed scrapie. Findings of this study suggest that orally exposed neonatal lambs of a susceptible (QQ) genotype exhibit a higher attack rate and shorter incubation period than older (4-month-old) lambs exposed to a larger dose (30x) of the same inoculum.