Scrapie Resistance Research Articles

Polymorphism of prion protein gene (PRNP) in Nigerian sheep

ABSTRACT Polymorphism of the prion protein gene (PRNP) gene determines an animal’s susceptibility to scrapie. Three polymorphisms at codons 136, 154, and 171 have been linked to classical scrapie susceptibility, although many variants of PRNP have been reported. However, no study has investigated scrapie susceptibility in Nigerian sheep from the drier agro-climate zones. In this study, we aimed to identify PRNP polymorphism in nucleotide sequences of 126 Nigerian sheep by comparing them with public available studies on scrapie-affected sheep. Further, we deployed Polyphen-2, PROVEAN, and AMYCO analyses to determine the structure changes produced by the non-synonymous SNPs. Nineteen (19) SNPs were found in Nigerian sheep with 14 being non-synonymous. Interestingly, one novel SNP (T718C) was identified. There was a significant difference (P < 0.05) in the allele frequencies of PRNP codon 154 between sheep in Italy and Nigeria. Based on the prediction by Polyphen-2, R154H was probably damaging while H171Q was benign. Contrarily, all SNPs were neutral via PROVEAN analysis while two haplotypes (HYKK and HDKK) had similar amyloid propensity of PRNP with resistance haplotype in Nigerian sheep. Our study provides valuable information that could be possibly adopted in programs targeted at breeding for scrapie resistance in sheep from tropical regions.

Evaluation of Resistance and Susceptibility to Prion Diseases in Pelibuey Sheep from Mexico

Abstract Transmissible spongiform encephalopathies (TSEs) are a group of neurodegenerative diseases with a chronic and fatal course, which are caused by a misfolded form of the cellular prion protein that is encoded by the host. The purpose of this work was to evaluate the resistance and genetic susceptibility to prion diseases in Pelibuey sheep from Mexico. The sequences of 99 Pelibuey sheep from the central and southwestern regions of Mexico were analyzed to determine the polymorphisms related to resistance and genetic susceptibility to scrapie, the Hardy-Weinberg equilibrium test and the D-Tajima test was used to identify the effect of evolutionary forces on the PRNP gene. Twelve non-synonymous polymorphisms Q101R, M112T, A116P, G127A, A136V, M137T, L141F, H143R, R154H, Q171R, Q171H, N176K were detected, in addition two synonymous substitutions 231R (agg/cgg) and 237L (ctc/ctg) were found. As a result of the sequence analysis, the ARR allele was not under the Hardy-Weinberg equilibrium, indicating that there is an evolutionary force at work, and the D-Tajima suggest the existence of purifying genetic selection. In conclusion, Pelibuey sheep exhibit genotypes for the PRNP gene that make them resistant to developing classic scrapie; at least 98% of the population is a carrier of an allele related to scrapie resistance; while for atypical scrapie there is a very high probability that an outbreak occurred in the herds, since the ARQ allele in combination with the L substitution at codon 141 confers susceptibility to carrier sheep.

Genetic Polymorphism at 15 Codons of the Prion Protein Gene in 156 Goats from Romania.

Background: The variability of prion protein gene (PRNP) codons and the frequency of alleles (K222, D146, and S146) that appear to confer genetic resistance to classical scrapie are still unknown in several goat populations/breeds prevalent in Romania. This work aims to assess the genetic polymorphism at 15 PRNP codons in Romanian goat populations to inform the development of goat breeding programs for scrapie resistance. Methods: Whole blood and hair follicles from Carpathian (50), French Alpine (53), and Banat’s White (53) breed goats were sampled to extract genomic DNA for genetic analyses and Sanger sequencing. In the targeted goat groups, one classical scrapie-positive Banat’s White goat was included. Results: The codons without polymorphisms were G37G, W102W, N146N, R151R, S173S, and I218I. The following non-synonymous polymorphisms of PRNP were recorded: P110P, P110S, P110T, T110T, G127G, G127S, I142I, I142M, T142I, H143H, P143P, R143R, R154R, H154R, P168P, Q168Q, Q211Q, Q211R, Q222Q, H222Q, K222K, S240S, P240P, P240S, and S240P. Conclusions: PRNP polymorphism was recorded in 60% (9/15) of codons. The scrapie-positive Banat’s White goat had G37G, W102W, T110T, G127G, I142I, H143H, N146N, R151R, R154R, P168P, S173S, R211R, I218I, Q222Q, and S240S. The K222 allele had a frequency of 6% (3/50) in Carpathian, 9.43% (5/53) in Banat’s White, and 15.09% (8/53) in French Alpine. Therefore, the polymorphisms detected in this sample of Romanian goat breeds are too rare to design a breeding program at the current time.

Prediction of Genetic Resistance for Scrapie in Ungenotyped Sheep Using a Linear Animal Model.

Selection based on scrapie genotypes could improve the genetic resistance for scrapie in sheep. However, in practice, few animals are genotyped. The objectives were to define numerical values of scrapie resistance genotypes and adjust for their non-additive genetic effect; evaluate prediction accuracy of ungenotyped animals using linear animal model; and predict and assess selection response based on estimated breeding values (EBV) of ungenotyped animals. The scrapie resistance (SR) was defined by ranking scrapie genotypes from low (0) to high (4) resistance based on genotype risk groups and was also adjusted for non-additive genetic effect of the haplotypes. Genotypes were simulated for 1,671,890 animals from pedigree. The simulated alleles were assigned to scrapie haplotypes in two scenarios of high (SRh) and low (SRl) resistance populations. A sample of 20,000 genotyped animals were used to predict ungenotyped using animal model. Prediction accuracies for ungenotyped animals for SRh and SRl were 0.60 and 0.54, and for allele content were from 0.41 to 0.71, respectively. Response to selection on SRh and SRl increased SR by 0.52 and 0.28, and on allele content from 0.13 to 0.50, respectively. In addition, the selected animals had large proportion of homozygous for the favorable haplotypes. Thus, pre-selection prior to genotyping could reduce genotyping costs for breeding programs. Using a linear animal model to predict SR makes better use of available information for the breeding programs.

Monitoring inbreeding and selection on scrapie resistance in a closed nucleus of Florina sheep breed

This study aims to investigate the impact of selection on scrapie resistance on the genetic variability of Florina sheep breed closed nucleus at the Research Institute of Animal Science, in the region of Central Macedonia, Greece. The complete pedigree dataset of the nucleus flock was used in the study, and the prion protein (PrP) gene polymorphisms were identified for each animal. Three annual selection regimes were simulated, where the selection of the rams and ewes for mating was based on the coancestry value between them and their PrP gene polymorphism. The population structure and pedigree were analyzed to assess the impact of selection on scrapie on the evolution of inbreeding and the effective population size.

Large-scale survey of prion protein genetic variability in scrapie disease-free goats from the United States.

Scrapie is a slowly progressive neurodegenerative disease of small ruminants caused by an accumulation of an abnormal isoform of prion protein in the central nervous system. Polymorphisms of the prion protein gene (PRNP) strongly modulate scrapie resistance and incubation period in goats. The aim of this study was to identify PRNP genetic variability in goats across the United States. Blood from a total of 6,029 apparent scrapie disease-free goats from 654 operations and 19 breeds were analyzed. Sequencing of PRNP revealed 26 genotypes with different rates based on eight codons. The GG127, RR154, and QQ222 genotypes were predominant and showed a remarkably high rate across all goats. The QK222 and NS146 genotypes, known to be protective against scrapie, were found in 0.6% [with 95% CI = (0.3, 1.2)] and 22.0% [95% CI = (19.1, 25.2)] of goats, respectively. The QK222 genotype was found in 23.1% of Oberhasli goats tested, with 95%CI = (3.9, 68.7)] and 22.0% of Toggenburg goats tested with 95%CI = (9.7, 42.5)], while NS146 was found in 65.5% of Savannah goats tested, with 95%CI = (30.8, 89.9), 36.7% of Boer goats tested, with 95%CI = (33.1, 40.4), 36.3% of Nubian goats tested, with 95%CI = (27.0, 46.7)], and 35.6% of LaMancha goats tested, with 95%CI = (22.8, 50.8%). The MM142 and IM142 genotypes were found more frequently in goats on dairy operations, while the HR143, NS146, and ND146 genotypes were found more frequently in goats on meat operations. Goats in the east region had a higher percentage of goats with RH154, RQ211, and QK222 genotypes than goats in the west region. The results of this study showed high genetic variability of PRNP among the U.S. goat population, with differences by location and breed, and may serve as a rationale for development of goat breeding programs at the national level to mitigate the risk of scrapie.

Scrapie Resistance Gene Identification using Optimized Taqman Test qPCR Method in Sheep on the Territory of the Republic of Serbia

Abstract Scrapie is an infectious neurodegenerative disease affecting the central nervous system of sheep and goats that belongs to transmissible spongiform encephalopathies. The disease is caused by the accumulation of proteinase-resistant isoform of the prion protein. The sheep predisposition to scrapie is associated with polymorphisms of the PrP gene. Genetic susceptibility to scrapie is mainly related to codons 136, 154, and 171. ARR sheep are strongly scrapie resistant and VRQ genotype is the most susceptible. Many countries have scrapie eradication programs based on using rams with resistant genotype. The eradication program has not yet been implemented in the Republic of Serbia. To examine the genetic makeup of sheep in Serbia related to scrapie, we optimized TaqMan probes of real-time polymerase chain reaction (qPCR) technique for three codons. Blood samples from 100 sheep were analyzed by qPCR and the majority of the examined sheep were AA homozygous for the 136 codon. For codon 154 the most frequent genotype was RR and for codon 171 the most frequent genotype was QQ.

PRNP Polymorphisms in Eight Local Goat Populations/Breeds from Central and Southern Italy.

Simple SummaryScrapie is a naturally occurring transmissible spongiform encephalopathy of sheep and goats. Polymorphisms in single nucleotides in the coding sequence of the prion protein gene play a major role in relative susceptibility or resistance to classical scrapie. The most recent modification of European Union Regulation 999/2001 (Regulation EU 772/2020) deals with the possible use of genetics in goats for scrapie outbreak management. In this work, sequence analysis of the prion protein gene was performed, with the main aim to obtain information about the genetic variability of eight Italian local populations/breeds and establishing phylogenetic relationships between the breeds/populations. This research could help to promote the adoption of selective breeding programs as a possible future strategy in scrapie control and outbreak prevention. In particular, genomic DNA was extracted from 219 goat whole blood samples and 13 polymorphic sites were observed, giving 24 different alleles. We observed the K222 allele, associated to scrapie resistance, with a mean frequency of 10%. Interestingly, we also described in Italy the circulation of the S146 allele: In particular, S146 only segregated in the Alpine breed at a frequency of 3.7%. This study adds information on genetic variability of the PRNP locus in goat populations/breeds, so contributing to the design of genetic control measure both in scrapie outbreaks and in disease prevention.In goats, as in sheep, genotypes of the prion protein gene (PRNP) can influence animals’ susceptibility to scrapie. Since the polymorphic codons in sheep are well known, a genetic selection plan has been implemented in Europe, in order to reduce the prevalence of susceptible genotypes to scrapie. In Italy, no breeding plan for scrapie resistance in goats has been adopted, yet. Likewise, according to the most recent modification of Regulation EU 999/2001 (Regulation EU 772/2020) of the European Commission (EU), based on all the available experimental and in field data, K222, D146 and S146 polymorphisms could be used as scrapie resistance alleles in genetic management both in scrapie outbreaks and in disease prevention. In order to collect data on the variability of PRNP, the present study aimed to analyze the sequence of the PRNP gene in eight Italian local goat populations/breeds reared in central and southern Italy (Bianca Monticellana, Capestrina, Facciuta della Valnerina, Fulva del Lazio, Garganica, Grigia Ciociara, Grigia Molisana, and Teramana), some of which were investigated for the first time; moreover, two cosmopolitan breeds (Alpine and Saanen) were included. Blood samples were collected from 219 goats. Genomic DNA was extracted from whole blood. DNA was used as template in PCR amplification of the entire PRNP open reading frame (ORF). Purified amplicons have been sequenced and aligned to Capra hircus PRNP. Particularly, the alleles carrying the resistance-related 222 K polymorphism occurred in all populations with a frequency between 2.5% and 12.5%. An additional resistance allele carrying the S146 variant was observed with a frequency of 3.7% only in the Alpine breed. For three of the estimated alleles, we could not establish if the found double polymorphisms in heterozygosis were in phase, due to technical limitations. In this context, in addition to selective culling in scrapie outbreaks according to the European regulation in force, in the future, selection plans could be adopted to deal with scrapie and to control its diffusion, meanwhile paying attention to preserve a high variability of PRNP.

Selection and drift reduce genetic variation for milk yield in Manech Tête Rousse dairy sheep

Decreases in genetic variance over generations reduce future genetic gain. We studied the evolution of genetic variance in the dairy sheep breed Manech Tête Rousse, which has been selected for increasingly complex objectives, including, in this order, milk yield, milk contents, scrapie resistance, and somatic cell score. We estimated base population genetic variance and genetic variance by sex and per year of birth from 1981 to 2014. The data consisted of 1,842,295 milk yield records (from 1978 to 2017) and a pedigree including 530,572 females (96% of them with records) and 3,798 artificial insemination males. As a measure of drift, we computed average relationships for each cohort from which we derived expected reduction of variance due to increased relationships. The difference between observed and expected reductions in genetic variances is the reduction in genetic variance due to selection. Average relationships increased steadily but slowly in both sexes. For females, genetic variance reduced with time until a plateau was reached at around 90% of the initial genetic variance. The reduction due to relationships (roughly 3% cumulated in 30 yr) was smaller than that due to selection (roughly 10% across the last years). A smaller loss due to selection was seen in recent years, possibly due to a change in selection objectives. These results agree well with theoretical expectations. The pattern of the evolution of genetic variance in males was similar to that for females but with a stronger reduction because of strong selection of AI males at birth. We conclude that the reductions in genetic variation due to selection and drift agree with expectations, and none of the reductions are very strong in this population because of control of inbreeding and smooth changes in selection objectives over time.

Genetic variability following selection for scrapie resistance in six autochthonous sheep breeds in the province of Bolzano (northern Italy).

Scrapie is an ovine transmissible spongiform encephalopathy, and its susceptibility is associated with polymorphisms in the prion protein gene (PRNP). Genetic selection is currently the most effective mean for eradication of the susceptible VRQ allele in favour of resistant ARR allele. Maintenance of genetic diversity should be one of the major objectives in breeding programmes, especially in endangered breeds, and genetic information are an excellent alternative to pedigree data where these information are missing. The aim of our study was to determine changes of genetic variability in six native sheep breeds from autonomous province of Bolzano, northern Italy, following simulation of scrapie selection scenarios. A total of 684 rams were investigated for PRNP polymorphisms and for 10 microsatellite loci to estimate genetic variability. Across all loci, a total of 163 alleles were detected with a mean of 10.4 alleles per locus. Average observed (Ho) and unbiased expected (uHe) heterozygosity overall loci were 0.74 and 0.78, respectively, showing a statistically significant deviation from Hardy-Weinberg equilibrium (HWE) in all breeds. This heterozygosity deficit was confirmed by a positive fixation index (Fis), determining a moderate inbreeding in each breed. Simulating a soft selection, where only rams having at least a VRQ allele should be excluded from reproduction, Ho, uHe and Fis values remained almost unchanged, indicating that genetic variability should not be affected by the removal of these individuals. With a mild selection scenario, considering only rams with at least one ARR allele, we observed a decrease in the mean alleles per breed (8.9) and the maintenance of heterozygosity deficiency, except for two breeds, where it was any longer significant. These results showed that selection strategies allowing use of heterozygous as well homozygous ARR rams might be the right compromise to improve resistance to scrapie and to do not dramatically affect genetic variability of these breeds.

Large-scale PRNP genotyping of small ruminants using an automated high-throughput MALDI-TOF MS assay.

Scrapie resistance or susceptibility in sheep is associated with single nucleotide polymorphisms (SNPs) at codons 136, 154, and 171 of the prion protein gene (PRNP). In addition, phenylalanine mutation at codon 141 has been recognized as a risk factor for atypical scrapie. In contrast, K222, D146, and S146 alleles confer genetic resistance to classical scrapie in goats. High-throughput genotyping technologies would provide significant benefits in scrapie eradication plans. The ability to resolve oligonucleotides varying in mass by less than a single nucleotide makes MALDI-TOF mass spectrometry (MS) a suitable platform for PRNP genotyping. We evaluated the commercial Myriapod scrapie kit (Diatech Pharmacogenetics), associated with a highly automated processing platform incorporating MALDI-TOF MS technology, to detect SNPs at codons 136, 154, 171, 141, and 222 of small ruminant PRNP. The Myriapod scrapie kit was accredited according to UNI CEI EN ISO/IEC 17025. We present the genotyping results of 10,960 sheep in Sicily and 1,822 goats in Sicily and Calabria (southern Italy) tested during 2017. We found a high frequency (43.9%) of the protective ARR allele in sheep and a promising 12.3% of the resistant K222 variant in goats. This efficient and high-throughput method is suitable for extensive PRNP genotyping, as requested in the European scrapie eradication plan.

Two novel amino acid substitutions in highly conserved regions of prion protein (PrP) and a high frequency of a scrapie protective variant in native Ethiopian goats

BackgroundPolymorphisms of the prion protein gene may influence scrapie susceptibility in small ruminants through modified protein conformation. At least 47 amino acid substitutions and 19 silent polymorphisms have been described in goat PRNP reported from several countries. The objective of this study was to investigate PRNP polymorphisms of native Ethiopian goat breeds and compare the results with other goat breeds.ResultsThe analysis of the prion protein gene PRNP in 229 goats belonging to three of the main Ethiopian native goat breeds showed a remarkably high frequency (> 34.6%) of p.(Asn146Ser) in these breeds, a variant involved in scrapie resistance in Cyprus. In addition, two novel amino-acid substitutions p.(Gly127Ala) and p.(Thr193Ile), with frequencies ranging from 1.5 to 7.3% were detected. Both amino acids are well conserved in prion proteins (PrP) of most species and these changes have never been reported before in goats worldwide. Residue 127 is within the N–terminal domain of PrP and is probably involved in the recruitment of neural cell adhesion molecules (NCAM). Residue 193 is within the highly conserved string of 4 threonines that plays a role in determining the efficiency of prion protein conversion towards its pathological form.ConclusionTwo novel coding polymorphisms and a high frequency of a scrapie protective variant indicate a high level of genetic diversity in PRNP of Ethiopian goats. This finding increases the interest in exploring PRNP polymorphisms of native goat breeds in areas where cross breeding with foreign goats has rarely occurred.

Impact of polymorphisms at the PRNP locus on the performance of dairy goats reared under low-input pastoral farming systems

Our hypothesis was that polymorphisms in the PRNP locus controlling animal resistance to scrapie do not adversely affect overall performance of dairy goats reared under low-input pastoral farming systems. Hence, the objective was to investigate the association of polymorphisms at codons 146 (N/S/D), 211 (R/Q) and 222 (Q/K) in the PRNP gene locus with milk production, udder health and body condition score (BCS) of dairy goats in Greece. A total of 766 goats from seven farms were used. Goats belonged to two indigenous Greek breeds, Eghoria (n = 264) and Skopelos (n = 287), and one foreign breed, Damascus (n = 215); the former two were treated together as one indigenous population. Nuclear DNA was extracted from individual blood samples. Polymorphisms at the studied codons were detected with Real-Time PCR analysis. Milk production, udder health and BCS of individual animals were recorded monthly for two consecutive milking periods. Milk production traits included daily and total milk yield and corresponding milk fat, protein, lactose and solids-non-fat (SNF) content and yield. Udder health traits included udder asymmetry, fibrosis, abscesses and total number of udder problems per milking period, milk somatic cell count, and total viable count. The allele substitution effect at codons 146, 211 and 222 on the studied traits was assessed with mixed linear models. Overall, resistance-associated alleles did not affect the studied traits. Nominally statistically significant effects on some milk composition, udder health and BCS traits were detected, which did not remain significant after correcting for multiple testing. In conclusion, genetic selection for scrapie resistance in dairy goats is not likely to compromise animal performance. Nevertheless, continuous monitoring of selective breeding programmes is advised to safeguard against possible emerging side-effects on animal performance and genetic diversity.

Genotyping of prion protein in black merino sheep from the Iberian Peninsula

The black merino variety is an emerging fine-wool merino breed variety of sheep. Their flocks have been reared in Portugal and Spain under different criteria regarding the selection for scrapie resistance haplotypes. This study was aimed to report for the first time in the black merino the prion protein (PRNP) haplotype frequency distribution using sequencing. A sample of 120 sheep belonging to four flocks was analyzed in this study. Three unselected flocks from Portugal and one selected for ARR/ARR genotypes from Spain. The number of polymorphism sites was extended here from three positions (136, 154 and 171) to five by adding the positions 141 and 143 due to both to show its current intermediate frequency in Portuguese sheep and to study the genetic relationship among haplotypes using networks analysis. In this way susceptibility to classical and atypical scrapie can be elucidated. Eight haplotypes could be reported in the black merino as follows: ALHRR, ALHRQ, ALRRQ, ALHHQ, AFHRQ, ALHRH, VLHRQ and T112ALHRQ but the last was a singleton haplotype. The results showed high genetic differentiation among flocks due to an unbalanced frequency distribution of haplotypes suggesting scarce genetic flow among them in Portugal. After collapse haplotypes to three positions (136, 154 and 171) the black merino from Portugal showed similar haplotype frequencies than in previous study except for the VRQ haplotypes with higher frequency in this study. The finding of AFHRQ was interesting because it was associated to atypical scrapie being the only cases declared in Portugal of this disease. In conclusion, we reported high richness in haplotypes in this sheep, but its distribution critically depends on the studied flock. This genetic richness enables to analyze different scenarios before implemented breeding programmes. In this way the application of a breeding programme to control scrapie appears to be a less challenging task.

Sheep With the Homozygous Lysine-171 Prion Protein Genotype Are Resistant to Classical Scrapie After Experimental Oronasal Inoculation.

Scrapie is a fatal neurodegenerative disease of sheep resulting from the accumulation of a misfolded form of the prion protein (PrPSc). Polymorphisms in the host prion protein gene ( PRNP) can affect susceptibility to the scrapie agent. Lysine (K) at codon 171 of PRNP is an inadequately characterized, naturally occurring polymorphism in sheep. We inoculated Barbado sheep with PRNP genotypes QQ171, QK171, or KK171 by either the intracranial (IC, n = 2-7 per genotype) or oronasal (ON, n = 5 per genotype) routes with a scrapie isolate to investigate the effect of lysine at codon 171 on susceptibility. When neurologic signs were observed or at the end of the experiment (70 months postinoculation [MPI]), sheep were necropsied and tissue collected for histopathologic, immunohistochemical, enzyme immunoassay and Western blot examination for PrPSc. All genotypes of sheep developed scrapie after IC inoculation. After ON inoculation, sheep with the QK171 genotype had prolonged incubation periods compared to the QQ genotype. During the experiment, 2 of 5 of the ON-inoculated QK genotype sheep developed neurologic signs and had PrPSc in the brain. The other 3 of 5 sheep were asymptomatic at 70 MPI but had detectable PrPSc in peripheral tissues. None of the ON-inoculated sheep of the KK171 genotype developed signs or had detectable PrPSc. Our experiments demonstrate that sheep with the KK171 genotype are resistant to scrapie via oronasal exposure and that sheep with the QK171 genotype have prolonged incubation relative to QQ171 sheep. The K171 prion protein allele may be useful to enhance scrapie resistance in certain breeds of sheep.

35 Classical Scrapie Resistance in Goats Based on S146

35 Classical Scrapie Resistance in Goats Based on S146

The response of prion genic variation to selection for scrapie resistance in Hungarian indigenous sheep breeds.

The authors studied the present status of Hungarian indigenous sheep breeds based on the genetic background of scrapie resistance. The aim of this investigation was to estimate the relative frequency of prion haplotypes, genotypes and risk categories, as well as to reveal the efficiency of the scrapie eradication programme achieved over the last decade. A novel approach in the characterisation of prion by using its genic variation was also implemented. The authors established that the proportion of deleterious sites (%) can be a useful indicator of the eradication programme. Based on a large sample size, it was confirmed that the scrapie resistance of the Cikta breed is low, and the classification of this breed according to risk category has not improved. However, the frequent genotype ARQ and risk category 3 can also be considered characteristic of the breed. The careful use of these genotypes is permitted and will contribute to the maintenance of breed diversity. The response of prion genic variation to selection for scrapie resistance in the other breeds (Tsigai, Milking Tsigai, White Racka, Black Racka and Gyimes Racka) was definitely successful.

In silico investigation of the prion protein glycosylation profiles in relation to scrapie disease resistance in domestic sheep (Ovis aries)

The prion protein is a membrane-bound glycoprotein which consists mainly α-helix structure. In contrast, the infectious prion protein shows the beta-sheet structure. The prion-associated diseases are all lethal neurodegenerative abnormalities, called transmissible spongiform encephalopathies. Scrapie is the most common type of these illnesses affecting sheep, goats, and moufflon. The VRQ, AHQ, ARR and N146S polymorphisms in the sheep prion gene have been found to be associated with resistance to scrapie disease. So far, the relationship of polymorphisms to three-dimensional protein structures, post-translational modifications, and scrapie resistance has not been studied. In this study, the potential N- and O-glycosylation positions of sheep prion protein polymorphisms were analyzed, the secondary and three-dimensional protein structure models were predicted, three-dimensional glycoprotein models were constructed and the role of glycosylation positions in protein interactions was investigated. Here, we found that protein secondary and three-dimensional structures vary among polymorphisms. Moreover, we found wild-type prion and all polymorphic variants show N-glycosylation at Asn184 and Asn200 positions, while O-glycosylation profiles are variant-specific. We also found that structural changes among prion polymorphisms leads to the formation of variant spesific O-glycosylation profiles and these positions are associated with protein interactions. Based on these findings, we suggest that O-glycosylation may be effective on resistance/susceptibility of sheep prion polymorphisms to scrapie disease.

Correction to: Biodiversity and selection for scrapie resistance in sheep: genetic polymorphism in eight breeds of Algeria

Correction to: Biodiversity and selection for scrapie resistance in sheep: genetic polymorphism in eight breeds of Algeria

Spatio-temporal and risk factor analysis of alleles related to Scrapie resistance in sheep in Great Britain before, during and after a national breeding program

Certain genotypes of sheep have been identified to increase their susceptibility (the VRQ allele) or resistance (the ARR allele) to classical scrapie. This study’s aim was to assess the spatio-temporal pattern of the ARR and VRQ alleles in Great Britain (GB) and to explore the risk factors associated to their presence.Data was collected from the GB scrapie active surveillance program, the sheep and goat inventory survey (GB census survey) and the agricultural survey for the period 2002-2015. Spatio-temporal trends of genotypes were assessed through the use of choropleth maps, spatial cluster and linear regression analyses. Multivariable mixed effect logistic regression analyses were performed to investigate the association between the resistant or susceptible genotypes, and breeds, farm purpose, animal purpose, surveillance stream, country location and herd size.The results show a significant upward trend in the frequency of most resistant ARR alleles (1.15% per year, 95%CI: 0.76-1.53) and significant downward trend of most susceptible VRQ alleles (-0.40% per year; 95%CI: -0.69 to -0.10]. The trend continues after the termination of the national scrapie plan in 2009. Breeds such as Herdwick (OR = 0,26; 95%CI: 0.14-0.46), Shetland (OR = 0.22; 95%CI: 0.13-0.39), Swaledale (OR = 0.58; 95%CI: 0.47-0.73), Scottish blackface (OR = 0.54; 95%CI: 0.41-0.71) and Welsh Montain (OR: 0.59; 95%CI: 0.44-0.79) were identified with lower odds ratios of having the resistant ARR allele, while Beulah speckled face (OR = 1.58; 95%CI: 1.04-2.41), Jacob (OR = 2.91; 95%CI: 1.33-6.40), Lleyn (OR = 2.94; 95%CI: 1.28-6.74) and Suffolk (OR = 2.19; 95%CI: 1.69-2.84) had higher odds ratios of having the ARR allele. Other risk factors associated to presence of ARR allele were finishing farms (OR = 1.15; 95%CI: 1.06-1.24) and farms in Scotland (OR = 0,78; 95%CI: 0.73-0.83) and in Lowland grazing areas (OR = 1.53; 95%CI: 1.39-1.67). Factors associated with presence the VRQ genotype were farms in Scotland (OR = 0,85; 95%CI: 0.77-0.93) and breeds such as Herdwick (OR = 2.2; 95%CI: 1.08-4.97), Shetland (OR = 4.12; 95%CI: 2.20-7.73) and Sweledale (OR = 1.51; 95%CI: 1.10-2.09). For the most resistant genotype, two significant spatial clusters were identified: a high-risk cluster in the south-west of GB (RR = 1.51, p < 0.001) and a low-risk cluster in northern GB (RR = 0.65, p < 0.001). For the most susceptible genotypes, one significant high-risk cluster was identified in Wales (RR = 2.89 and p = 0.013).Surveillance for classical scrapie could be improved with a risk-based approach by focussing on those areas and farm types identified to have higher frequency of VRQ alleles and less frequency of ARR alleles. Scrapie control strategies could focus on developing breeding programs on farms with Shetland, Herdwick and Swaledale breeds.