Incidence Of Scrapie Research Articles

Analysis of differences in prion protein gene (PRNP) polymorphisms between Algerian and Southern Italy's goats

Goats are adapted to live in harsh areas of the world ensuring minimum levels of milk and meat production without much economic input. Scrapie is a fatal prion disease of small ruminants detected in different countries worldwide. Polymorphisms in the prion protein gene (PRNP) modify the degree of susceptibility/resistance to scrapie. PRNP polymorphisms were analysed in the four main Algerian goat breeds (Naine de Kabylie, Arbia, Mozabite, Mekatia), and in two native breeds of South Italy (Aspromontana from Calabria and Cilentana from Campania). The results were compared also with two previously described Sicilian native breeds (Girgentana and Rossa Mediterranea). Seven amino acid substitutions were detected in Naine de Kabylie goat which represents the original native Berber breed. All other breeds presented no more than six variants and common polymorphisms were present at codons, 154 and 240. Isoleucine at 137 was present in Algerian breeds only. The Italian Cilentana shared more variants with Algerian breeds whereas the Aspromontana breed was the only one to show a Serine at 127. Exclusive new variants were not detected since all polymorphisms were already described in other goats worldwide. The scrapie protective allele encoding lysine (K) at codon 222 was detected in the Naine de Kabylie and M’zabite breeds at low-frequency whereas is present with frequencies higher than 10% in all the Italian breeds reared in regions with elevated scrapie incidence. The overall results showed a substantial number of polymorphisms in PRNP, particularly in Naine de Kabylie breed, which carried also unique genotypes.

A cross-sectional study of PRNP gene in two native Sicilian goat populations in Italy: a relation between prion gene polymorphisms and scrapie incidence.

Transmissible spongiform encephalopathies (TSEs) are a group of neurodegenerative diseases affecting humans and animals, and scrapie in small ruminants is considered the archetype of TSEs. Derivata di Siria is a native dairy goat of Sicily (south Italy), which is related to Syrian goat breeds. Scrapie disease is considered endemic in Sicily since 1997, following the administration of an infected vaccine.Derivata di Siria goatswere involved in six of 66 scrapie-infected flocks in Sicily. Prion protein gene (PRNP) analysis revealed that none of the scrapie cases carried the p.Gln222Lys variant. Sequencing of PRNP in this goat population showed a high frequency (15%) of p.Gln222Lys variant confirming its association with scrapie resistance. PRNP polymorphisms were also analysed in the population of Pantelleria, a small Sicilian Island, where scrapie has never been reported. The native goat breed 'Pantesca' was maintained up to almost 80 years and the size of the sheep population on this island has historically been very low. Currently, a crossbreed goat population of 253 heads is present on the island. PRNP genotyping of Pantelleria goats showed genetic variation, with low presence of wild-type goats and the lack of protective alleles. These data reinforce the association between PRNP polymorphisms in small ruminants and scrapie incidence.

Scrapie incidence and PRNP polymorphisms: rare small ruminant breeds of Sicily with TSE protecting genetic reservoirs.

BackgroundTransmissible spongiform encephalopathies (TSE) are fatal neurodegenerative diseases of several mammalian species, including humans. In Italy, the active surveillance through rapid tests on brain stem from small ruminants started in 2002 on randomly selected samples of healthy slaughtered animals. Sampling number was proportionally related to the regional small ruminant population. Of the twenty Italian regions, Sicily has the second largest population of small ruminants which is mainly constituted by crossbreed animals (>70 %). Sicily contains also three native sheep breeds Pinzirita, Comisana and Valle del Belice. Native goat breeds are Girgentana, Messinese, Argentata dell’Etna, Maltese and Rossa Mediterranea. The polymorphisms of prion protein gene (PRNP) may influence disease susceptibility and breeding programs for genetic TSE resistance are being applied in sheep. Protective alleles have been recently reported for goats also. These differ from those in sheep and may allow breeding programs in the near future.In this paper the data of active surveillance for scrapie control in general population of small ruminants in Sicily are reported together with the analysis on the polymorphism of PRNP in a number of Sicilian autochthonous breeds. The evaluation of the frequency of protective alleles is fundamental for the implementation of a TSE resistance breeding program.ResultsTSE surveillance in small ruminants in Sicily showed a of total fifty seven scrapie outbreaks from 1997 to 2014 involving mainly crossbreed animals. The PRNP polymorphism analysis in autochthonous breeds showed protective allele frequencies of 30–40 % ARR in sheep and 12–18 % K222 in three of the four goat breeds; these breeds are distributed over limited areas of the island.ConclusionThe study on PRNP polymorphisms in Sicilian small ruminant population showed higher frequency of the protective alleles compared to most other European breeds. Our results suggest that PRNP genetic variety in Sicilian sheep and goats can be a resource for TSE resistance breeding programmes while maintaining the conservation of endangered breeds and valorisation of their typical food products.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-016-0766-9) contains supplementary material, which is available to authorized users.

A relevant long-term impact of the circulation of a potentially contaminated vaccine on the distribution of scrapie in Italy. Results from a retrospective cohort study

A sudden increase in the incidence of scrapie in Italy in 1997 was subsequently linked to the use of a potentially infected vaccine against contagious agalactia. The relative risk for the exposed farms ranged between 6 and 40. The aim of this study was to assess the long-term impact of exposure to the potentially scrapie-contaminated vaccine on the Italian classical scrapie epidemic. We carried out a retrospective cohort study, fitting mixed-effects Poisson regression models, dividing national geographic areas into exposure categories on the basis of the vaccine circulation levels. We took into account the sensitivity of the surveillance system applied in the different areas. The population attributable fraction (PAF) was used to assess the impact on the total population of farms associated with the effect of circulation of the vaccine. The provinces where the vaccine was more often sold were noted to have a higher level of disease when compared to those provinces where the vaccine was sold less often (incidence rate ratio [IRR]: 2.7; 95% confidence interval [CI]: 1.1-6.5). The population attributable fraction was high (68.4%). Standardization techniques allowed to account for the potential of geographical variability in the sensitivity of the Italian surveillance system. Although the number of the directly exposed farms was limited, an important long-term impact of the vaccine circulation could be quantified in terms of secondary outbreaks likely due to the exchange of animals from directly exposed flocks.

Factors influencing temporal variation of scrapie incidence within a closed Suffolk sheep flock

Several studies have shown that transmission of natural scrapie can occur vertically and horizontally, and that variations in scrapie incidence between and within infected flocks are mostly due to differences in the proportion of sheep with susceptible and resistant PRNP genotypes. This report presents the results of a 12-year period of scrapie monitoring in a closed flock of Suffolk sheep, in which only animals of the ARQ/ARQ genotype developed disease. Among a total of 120 of these, scrapie attack rates varied between birth cohorts from 62.5 % (5/8) to 100 % (9/9), and the incidence of clinical disease among infected sheep from 88.9 % (8/9) to 100 % (in five birth cohorts). Susceptible sheep born to scrapie-infected ewes showed a slightly higher risk of becoming infected (97.2 %), produced earlier biopsy-positive results (mean 354 days) and developed disease at a younger age (median 736 days) than those born to non-infected dams (80.3 %, 451 and 782 days, respectively). Taken together, this was interpreted as evidence of maternal transmission. However, it was also observed that, for the birth cohorts with the highest incidence of scrapie (90-100 %), sheep born to infected and non-infected dams had a similar risk of developing scrapie (97.1 and 95.3 %, respectively). Compared with moderate-attack-rate cohorts (62.5-66.7 %), high-incidence cohorts had greater numbers of susceptible lambs born to infected ewes, suggesting that increased rates of horizontal transmission in these cohorts could have been due to high levels of environmental contamination caused by infected placentas.

Caprine PRNP polymorphisms at codons 171, 211, 222 and 240 in a Greek herd and their association with classical scrapie

The association between PRNP variation and scrapie incidence was investigated in a highly affected Greek goat herd. Four mutations were identified at codons 171Q/R, 211R/Q, 222Q/K and 240P/S. Lysine at codon 222 was found to be associated with the protection from natural scrapie (P=0.0111). Glutamine at codon 211 was observed in eight animals, all of them being scrapie-negative, indicating a possible protective role of this polymorphism although statistical analysis failed to support it (P=0.1074). A positive association (P=0.0457) between scrapie-affected goats and the wild-type Q(171)R(211)Q(222)S(240) allele is presented for the first time. In addition, a novel R(171)RQS allele, which is identical to the A(136)R(154)R(171) allele that has been associated with resistance to classical scrapie in sheep, was observed in low frequency. Resistant alleles that include K(222) and Q(211) are absent or rare in sheep and can provide the basis for the development of a feasible breeding programme for scrapie eradication in goats.

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 impact of sheep breed on the risk of classical scrapie

The risk of classical scrapie in sheep is associated with polymorphisms in the prion protein (PrP) gene. In recent years, large-scale selective breeding programmes for sheep at lower risk of disease have been undertaken across the European Union. We analysed large-scale datasets on scrapie and sheep demography to investigate additional effects of sheep breed on scrapie risk. There was evidence for variation between certain breeds in the scrapie risk of some PrP genotypes, which could be caused by innate breed differences or distinct scrapie strains circulating within them. While the PrP genotypes of cases are generally consistent across breeds, some exceptions provide evidence that scrapie strain may influence affected PrP genotypes to a greater extent than innate breed differences. There was a significant association between the breed-level incidence of scrapie and the frequency of susceptible PrP genotypes in breeds. Our results lend support to selective breeding programmes which aim to reduce the frequency of high-risk PrP genotypes with measures not varying by sheep breed.

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.

PrP polymorphisms in Brazilian sheep

Ovine scrapie, a fatal, neurodegenerative disorder of the transmissible spongiform encephalopathies group is an emerging disease in Brazilian sheep. The disorder is due to the deposition of a conformationally altered form (PrP Sc) of the normal prion protein (PrP C) in the central nervous and lymphoreticular systems. Mutations at codons 171 (Q, R, H, or K) and 136 (V or A) in the ovine prion gene (PrP), influence scrapie incidence, the codon 171 being the major determinant of susceptibility. This paper analyzed 301 sheep of Hampshire Down and Suffolk breeds in relation to codon 136 and 171. Genomic DNA was obtained from blood and genotypes were screened using polimerase chain reaction and restriction endonuclease digestion. The AA and QQ genotypes were the most frequent in Suffolk breed, while AA and QR were the more common in H. Down sheep. Considering both codons simultaneously the AAQQ genotype was the most frequent in Suffolk and AAQR in H. Down. Up to 90% of the animals had medium to high risk of developing the disease. This data could be used as a guide for selection procedures to reduce the risk of scrapie in Brazilian sheep flocks.

Potential Role of Soil in the Transmission of Prion Disease

Transmissible spongiform encephalopathies (TSE), or prion diseases, are a family of inevitably fatal neurodegenerative disorders affecting a variety of mammalian species. These diseases include scrapie in sheep and goats; bovine spongiform encephalopathy (BSE, “mad cow” disease) in cattle; chronic wasting disease (CWD) in North American deer, elk and moose; transmissible mink encephalopathy; and Creutzfeldt-Jakob disease (CJD, sporadic, familial and variant forms) and kuru in humans. These diseases are characterized by long incubation periods, spongiform degeneration of the brain and accumulation of an abnormally folded isoform of the prion protein, designated PrPSc, in brain tissue (Prusiner 1998). CWD and scrapie are the only TSEs that appear to be environmentally transmitted. Therefore, this chapter focuses primarily on these two TSEs. Scrapie has been known in sheep for at least 250 years (McGowan 1922). Most clinically infected sheep exhibit the obvious feature of excessive rubbing and scratching of the skin; the term “scrapie” derives from this symptom. The peak incidence of scrapie occurs in sheep three to four years of age although the earliest cases are seen at 18 months and the latest in animals older than 10 years (Dickinson 1976). The origin of the scrapie agent is unknown, but a familial pattern exists in natural sheep scrapie suggesting that genetics and, possibly, vertical transmission are important. Scrapie has a world-wide distribution and has been documented wherever sheep are raised, with the exception of Australia and New Zealand. CWD was first identified at a Colorado research facility in 1967 (Williams and Young 1980) and has since been identified in captive cervid populations in Wyoming, Wisconsin, Saskatchewan, South Dakota, Oklahoma, New York, Nebraska, Montana, Minnesota, Kansas and Alberta (Fig. 1⇓). In the free-ranging cervid population, CWD has been found in Wyoming, Wisconsin, Utah, South Dakota, Saskatchewan, New Mexico, Nebraska, Illinois, …

Breeding programmes for TSE resistance in British sheep: II. Assessing the impact on the prevalence and incidence of scrapie

By establishing a breeding programme for transmissible spongiform encephalopathie (TSE) resistance, there are plans to eradicate sheep TSEs from member states of the European Union (EU). In this paper, we used a simple age- and genotype-structured model to assess the impact of four breeding strategies on the prevalence and incidence of scrapie in the British sheep flock. The strategies ranged from the minimum EU requirements to compulsory implementation of the current National Scrapie Plan for Great Britain (NSP). All four strategies were predicted to reduce the prevalence and incidence of disease, though there was likely to be a delay of several years between the implementation of a breeding programme and the reduction in incidence. There were differences in the efficacy of the strategies, with the most stringent resulting in the greatest reduction in prevalence and incidence. However, the magnitude of the differences was not great, largely because all four strategies eliminated the VRQ allele, which is associated with a markedly higher risk of disease than any of the other alleles. Sensitivity analyses indicated that the model results were robust to selection bias when estimating the risk of infection; and that the efficacy of a breeding programme was unlikely to be compromised, unless the risk of infection is substantially underestimated by data on clinical disease.

Epidemiological analysis of data for scrapie in Great Britain.

In recent years, the control or eradication of scrapie and any other transmissible spongiform encephalopathies (TSEs) possibly circulating in the sheep population has become a priority in Britain and elsewhere in Europe. A better understanding of the epidemiology of scrapie would greatly aid the development and evaluation of control and eradication strategies. Here we bound the range of key epidemiological parameters using a combination of relatively detailed pathogenesis and demography data, more limited data on susceptibility and incubation times, and recent survey data on scrapie incidence in Great Britain. These data are simultaneously analysed using mathematical models describing scrapie transmission between sheep and between flocks. Our analysis suggests that occurrence of scrapie in a flock typically provokes changes in flock management that promote termination of the outbreak, such as the adoption of selective breeding, and that a large fraction of cases (possibly over 80%) goes undetected. We show that the data analysed are consistent with the within-flock reproduction number of scrapie lying in the range 1.5-6, consistent with previous epidemiological studies.

High incidence of scrapie induced by repeated injections of subinfectious prion doses.

To clarify the mechanisms leading to the development of Creutzfeldt-Jakob disease in some recipients of pituitary-derived human growth hormone (hGH), we investigated the effects of repeated injections of low prion doses in mice. The injections were performed, as in hGH-treated children, by a peripheral route at short intervals and for an extended period. Twelve groups of 24 mice were intraperitoneally inoculated one, two, or five times per week for 200 days with 2 x 10(-5) to 2 x 10(-8) dilutions of brain homogenate containing the mouse-adapted C506M3 scrapie strain. Sixteen control mice were injected once a week for 200 days with a 2 x 10(-4) dilution of normal brain homogenate. Of mice injected in a single challenge with a scrapie inoculum of a 2 x 10(-4), 2 x 10(-5), or 2 x 10(-6) dilution, 2/10, 1/10, and 0/10 animals developed scrapie, respectively. Control mice remained healthy. One hundred thirty-five of 135 mice injected with repeated prion doses of a 2 x 10(-5) or 2 x 10(-6) dilution succumbed to scrapie. Of mice injected with repeated scrapie doses of a 2 x 10(-7) or 2 x 10(-8) dilution, 52/59 and 38/67 animals died of scrapie, respectively. A high incidence of scrapie was observed in mice receiving repeated doses at low infectivity, whereas there was no disease in mice that were injected once with the same doses. Repeated injections of low prion doses thus constitute a risk for development of prion disease even if the same total dose inoculated in a single challenge does not induce the disease.

New ovine PrP gene haplotypes as a result of single nucleotide polymorphisms in the PrP gene promoter

Incidence of scrapie in sheep is strongly associated with PrP gene amino acid codon variants at positions 136, 154 and 171. However, there are breed differences in disease linkage and anomalous disease patterns which cannot obviously be explained by the '3 codon' genotype. Mouse studies indicate that PrP protein levels can influence scrapie disease progression and this prompted us to study the sheep PrP gene promoter region in a search for novel polymorphisms which may influence gene expression and hence disease susceptibility. The incidence of three single nucleotide polymorphisms (SNP) at positions C/A-5354, T/C-5382 and C/G-5622 within the PrP gene promoter region was determined from Neuropathogenesis Unit (NPU) and New Zealand (NZ) Cheviot and UK and NZ Suffolk sheep. The SNP variants A-5354 and G-5622 created consensus sequences for STAT and SP1 transcription factors, respectively, and C-5382 was within Motif 1, one of four conserved motifs found within the promoter region of mammalian PrP genes. The occurrence of C/A-5354 and T/C-5384 SNP exhibited differential associations with the PrP open reading frame (ORF) variants linked to scrapie susceptibility. A significant imbalance in the incidence of the C-5354/AXQ haplotype was found in the NPU Cheviot flock. C-5382 was not found in Suffolk sheep of either UK or NZ origin. The G-5622 SNP was found at a lower incidence in Suffolk sheep compared with Cheviots. The range of transcription factor binding motif profiles in the PrP gene promoter may act to modulate PrP gene activity and warrants further large-scale study.

The incidence of genotypes at codon 171 of the prion protein gene (PRNP) in five breeds of sheep and production traits of ewes associated with those genotypes1,2

Scrapie is one of several transmissible spongiform encephalopathies of livestock. Disease susceptibility is linked to polymorphisms in the normal prion protein gene that encodes the mammalian prion precursor. Codon 171 of this gene is a major determinant of scrapie susceptibility. Selection for arginine (R) at codon 171 is encouraged by the USDA to decrease the incidence of scrapie. Objectives of this study were to determine the frequency of R allele variants at codon 171 in a sample of sheep from five breeds (Columbia, Hampshire, Rambouillet, Suffolk, and Targhee) and western white-faced commercial ewes and to determine whether the R allele is associated with ewe and lamb production traits. Genotyping was performed on 532 ewes and 901 lambs from the University of Wyoming flock, in addition to 820 rams from 52 sheep producers from Wyoming and surrounding areas, using a DNA mismatch assay that discriminated the R allele from others at codon 171. Genotyping was performed by DNA sequencing on 127 rams representing all breeds, except Hampshire from the USDA Sheep Experiment Station at Dubois, ID. The 171R allele was found in all five breeds and in the commercial western white-faced ewes. Genotype frequencies varied (P < 0.001) by breed in ewe and ram populations. Influence of R-allele frequency on ewe lambing records and individual lamb records was analyzed for Columbia (62, 161, 121), Hampshire (89, 193, 162), Rambouillet (87, 179, 133), Suffolk (67, 178, 161), and commercial sheep (227, 463, 324) for numbers of ewes, total number of ewe production records, and individual lamb records, respectively. Suffolk ewes without the R allele (non-R/non-R) gave birth to more (P <or= 0.005) multiple lambs than heterozygous non-R/R ewes. Homozygous non-R/non-R Suffolk ewes weaned lighter (P = 0.02) individual lambs but more (P = 0.03) total weight of lamb than heterozygous non-R/R ewes. Although ewes genotyped homozygous non-R/non-R or heterozygous non-R/R at codon 171 from the commercial flock gave birth to more (P <or= 0.002) multiple lambs than ewes genotyped R/R, differences were not detected (P = 0.14) in total weight of lamb weaned. Production traits of Columbia, Hampshire, and Rambouillet ewes did not differ (P >or= 0.08) by ewe genotype. Lamb birth and weaning weights were not influenced (P >or= 0.12) by lamb genotype in any of the breeds or in the commercial flock. In this population, ultimate lamb production was only influenced by genotype at codon 171 in the Suffolk flock.

Demonstration of lateral transmission of scrapie between sheep kept under natural conditions using lymphoid tissue biopsy

Scrapie free adult sheep were introduced to a sheep flock specifically maintained to maximise scrapie infection. Native born sheep of the highly susceptible VRQ/VRQ genotype in this flock show highly efficient transmission, evidenced by 100% infection, with an age at death of less than 2 years. Infection in introduced sheep was identified by biopsy of tonsilar and nictitating membrane lymphoid tissue. Progeny of these sheep were monitored and clinical disease confirmed by examination of the brain using routine diagnostic methods. Naïve sheep of New Zealand origin introduced to the flock in adulthood became infected, demonstrating that lateral transmission had occurred. Lambs born to introduced ewes became infected and died at the same age as lambs born to native ewes, consistent with lateral transmission of scrapie to lambs. Although maternal transmission cannot be totally excluded for the lambs in this study, the data are consistent with lateral transmission being the most important means of spread leading to the high incidence of scrapie observed in this flock.

Multiplicative genetic effects in scrapie disease susceptibility

Despite experimental evidence that scrapie is an infectious disease of sheep, variations of the occurrence of the natural disease suggest an influence of host genetic factors. It has been established that the genetic polymorphism of the prion protein (PrP) gene is correlated to the incidence of scrapie and to the survival time: five polymorphisms have been described by variations at amino-acid codons 136, 154 and 171. In this paper we study the effect on scrapie susceptibility of the pairing of the five allelic variants known to exist: we show that scrapie susceptibility is given by the produce of the elementary allelic factors. This first well-documented evidence of a multiplicative property of genetic risk factors could give hints on the underlying mechanisms of prion-induced neurodegenerative diseases.

Evidence for the transmission of scrapie to sheep and goats from a vaccine against Mycoplasma agalactiae

An accidental infection from a vaccine was suggested as the explanation for the sudden increase in outbreaks of scrapie in Italy in 1997 and 1998. This paper describes a recent...

Scrapie in Britain during the BSE years.

The experimental transmission of bovine spongiform encephalopathy (BSE) to sheep1 raised the possibility that some sheep in the United Kingdom could have been infected during the 1980s after exposure to BSE-contaminated feed. In contrast to new diseases that have appeared in a number of feline species and wild ungulates2, the symptoms of BSE in sheep are very similar to another transmissible spongiform encephalopathy called scrapie, which has been endemic in Britain for over 200 years. Although so far no cases of BSE in sheep have been found, these may have been misdiagnosed as scrapie. Here we present data describing the historical changes in scrapie incidence3, and find no evidence for a peak in scrapie incidence before, during or after the BSE outbreak, making it unlikely that a substantial epidemic of BSE has occurred in the sheep population.