Scrapie Resistance Research Articles

Genetic profile of scrapie codons 146, 211 and 222 in the PRNP gene locus in three breeds of dairy goats.

Polymorphisms at PRNP gene locus have been associated with resistance against classical scrapie in goats. Genetic selection on this gene within appropriate breeding programs may contribute to the control of the disease. The present study characterized the genetic profile of codons 146, 211 and 222 in three dairy goat breeds in Greece. A total of 766 dairy goats from seven farms were used. Animals belonged to two indigenous Greek, Eghoria (n = 264) and Skopelos (n = 287) and a foreign breed, Damascus (n = 215). Genomic DNA was extracted from blood samples from individual animals. Polymorphisms were detected in these codons using Real-Time PCR analysis and four different Custom TaqMan® SNP Genotyping Assays. Genotypic, allelic and haplotypic frequencies were calculated based on individual animal genotypes. Chi-square tests were used to examine Hardy-Weinberg equilibrium state and compare genotypic distribution across breeds. Genetic distances among the three breeds, and between these and 30 breeds reared in other countries were estimated based on haplotypic frequencies using fixation index FST with Arlequin v3.1 software; a Neighbor-Joining tree was created using PHYLIP package v3.695. Level of statistical significance was set at P = 0.01. All scrapie resistance-associated alleles (146S, 146D, 211Q and 222K) were detected in the studied population. Significant frequency differences were observed between the indigenous Greek and Damascus breeds. Alleles 222K and 146S had the highest frequency in the two indigenous and the Damascus breed, respectively (ca. 6.0%). The studied breeds shared similar haplotypic frequencies with most South Italian and Turkish breeds but differed significantly from North-Western European, Far East and some USA goat breeds. Results suggest there is adequate variation in the PRNP gene locus to support breeding programs for enhanced scrapie resistance in goats reared in Greece. Genetic comparisons among goat breeds indicate that separate breeding programs should apply to the two indigenous and the imported Damascus breeds.

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

Scrapie is a prion disease that affects the sheep and goats. It belongs to the group of transmissible spongiform encephalopathies (TSE). TSEs are characterized by the accumulation of the pathological form (PrPSc) of the cellular prion protein (PrPC). The susceptibility of sheep to scrapie is influenced by polymorphisms in the PrP gene (PRNP). The aim of this study was to identify the genetic variability of sheep PRNP in Algerian sheep. Two-hundred and thirteen Algerian sheep from eight breeds (Ouled Djellal, Rembi, Hamra, Berbere, Barbarine, Sidaou, Taadmit and Tazegzawt) with no clinical manifestation of scrapie were analysed. Sequencing of the entire coding sequence of PRNP showed four main alleles (ARQ, ARR, AHQ and ARH) based on codons 136, 154 and 171 with different frequencies among the investigated breeds. Moreover, 14 additional nonsynonymous polymorphisms (Q101R, N103K, M112T, A116P, M137I, L141F, I142M, H143R, N146S, R151G, Y172D, N176K, H180Y and S240P) as well as two synonymous polymorphisms at codons 231 and 237 were found in the PRNP gene. Interestingly, the N103K, M137I and I142M polymorphisms were not described in sheep. The ARQ, ARR and ARH haplotypes were present in all breeds with a highest frequency of ARQ in Barbarine. The ARH was absent in Barbarine breed and the VRQ haplotype was absent in all Algerian breeds studied. The ARQ and ARR alleles were the most common with frequencies ranging from 30 to 65% and from 8 to 26%, respectively, in different breeds. These results represent the first study on PRNP variability in Algerian sheep and may serve as a basis for the development of breeding programmes to render national sheep breeds resistant to scrapie.

Predicting the impact of selection for scrapie resistance on PRNP genotype frequencies in goats

The European Union has implemented breeding programmes to increase scrapie resistance in sheep. A similar approach can be applied also in goats since the K222 allele provides a level of resistance equivalent to that of ARR in sheep. The European Food Safety Authority stated that breeding for resistance could be offered as an option for Member States to control classical scrapie in goats. We assessed the impact of different breeding strategies on PRNP genotype frequencies using a mathematical model that describes in detail the evolution of K222 in two goat breeds, Chamois Coloured and Saanen. Different patterns of age structure and replacement rate were modelled as factors affecting response to selection. Breeding for scrapie resistance can be implemented in goats, even though the initial K222 frequencies in these breeds are not particularly favourable and the rate at which the resistant animals increase, both breeding and slaughtered for meat production, is slow. If the goal is not to achieve the fixation of resistance allele, it is advisable to carry out selection only until a desired frequency of K222-carriers has been attained. Nucleus selection vs. selection on the overall populations is less expensive but takes longer to reach the desired output. The programme performed on the two goat breeds serves as a model of the response the selection could have in other breeds that show different initial frequencies and population structure. In this respect, the model has a general applicability.

The association between 171 K polymorphism and resistance against scrapie affection in Greek dairy sheep

Polymorphisms at codon 171 of the sheep prion protein gene (PRNP) are known to be genetically associated with scrapie (Hunter et al., 1993, 1997). Glutamine (Q) and histidine (H) at codon 171 contribute to scrapie susceptibility, while arginine (R) is associated with resistance. In some breeds, lysine (K) occurs at codon 171 (171 K), but its effect on scrapie resistance has not been fully determined yet. To investigate the role of 171 K in dairy sheep in Greece, we initially analyzed blood samples from 2368 sheep with no clinical signs of the disease, from thirteen scrapie-affected flocks that were then monitored for 5 years for scrapie clinical signs. During this period 177 sheep resulted positive in classical scrapie, after obex testing. The 177 scrapie-affected brain samples PRNP polymorphisms were analyzed using a Real Time-PCR protocol while the 2368 sheep blood samples were analyzed by a micro-array method based on primer extension (Homogenous Mass Extend Assay – SEQUENOM, Genaissance Pharmaceuticals) in MALDI-TOF-MS. Single Nucleotide Polymorphisms (SNPs) present at PRNP codons 136A/V (Alanine/Valine), 154R/H and 171Q/R/H/K were detected. The ARQ haplotype as well as the ARQ/ARQ genotype predominated. In the studied population, ARK haplotype was found at the highest frequency reported so far (6.98%) whereas, ARK/-genotypes had a total frequency of 12.81%. Relative risk (RR) and the percent relative effect were calculated to assess the association between ARK/-genotypes and scrapie affection. The results showed that carriers of K allele at codon 171 compared to Q and H alleles had ca. 0.25 and 0.40 times the risk of scrapie affection, respectively, indicating a possible association of K allele at codon 171 with scrapie resistance.

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.

Genetic diversity of the prion protein gene (PRNP) coding sequence in Czech sheep and evaluation of the national breeding programme for resistance to scrapie in the Czech Republic

Of 34 breeds kept in the Czech Republic 45,604 sheep were genotyped for codons 136, 154 and 171 in the prion protein gene (PRNP) during the years 2006-2014. In this cohort, haplotypes ARR, ARQ, ARH, AHQ, VRQ, AHR and ARK were detected. The haplotype AF141RQ associated with susceptibility to atypical scrapie was observed in nine out of 30 breeds analysed for this purpose. In addition, six rare nonsynonymous substitutions producing haplotypes AT137RQ, AN138RQ, AG151RQ, AH151RQ, ARL168Q and ARQE175 were identified in various breeds. Due to their low frequencies, these polymorphisms are of no potential importance for the breeding programme. With regard to their genetic particularity, Sumavka, Valachian and Cameroon breeds were screened for additional polymorphisms. Further haplotypes, AR143RQ and AS146RQ, were found in Sumavka and Cameroon, and in Valachian sheep, respectively. Frequencies of the ARR (resistance-associated), VRQ (susceptibility-associated) haplotypes, and of the most resistant ARR/ARR genotype calculated for sheep born in the years 2001-2003 and 2011-2013 documented effects of the 10year-lasting national breeding programme. The total frequency of ARR doubled from 36.8 to 75.8%, while the frequency of VRQ decreased from 4 to 0.7%. The total frequency of the ARR/ARR genotype increased from 17.7 to 59%. These data show that the national scrapie resistance breeding programme has had an important desirable effect on haplotype and genotype frequencies of PRNP in Czech sheep.

Variation in the prion protein sequence in Dutch goat breeds.

Scrapie is a neurodegenerative disease occurring in goats and sheep. Several haplotypes of the prion protein increase resistance to scrapie infection and may be used in selective breeding to help eradicate scrapie. In this study, frequencies of the allelic variants of the PrP gene are determined for six goat breeds in the Netherlands. Overall frequencies in Dutch goats were determined from 768 brain tissue samples in 2005, 766 in 2008 and 300 in 2012, derived from random sampling for the national scrapie surveillance without knowledge of the breed. Breed specific frequencies were determined in the winter 2013/2014 by sampling 300 breeding animals from the main breeders of the different breeds. Detailed analysis of the scrapie-resistant K222 haplotype was carried out in 2014 for 220 Dutch Toggenburger goats and in 2015 for 942 goats from the Saanen derived White Goat breed. Nine haplotypes were identified in the Dutch breeds. Frequencies for non-wild type haplotypes were generally low. Exception was the K222 haplotype in the Dutch Toggenburger (29%) and the S146 haplotype in the Nubian and Boer breeds (respectively 7 and 31%). The frequency of the K222 haplotype in the Toggenburger was higher than for any other breed reported in literature, while for the White Goat breed it was with 3.1% similar to frequencies of other Saanen or Saanen derived breeds. Further evidence was found for the existence of two M142 haplotypes, M142 /S240 and M142 /P240 . Breeds vary in haplotype frequencies but frequencies of resistant genotypes are generally low and consequently selective breeding for scrapie resistance can only be slow but will benefit from animals identified in this study. The unexpectedly high frequency of the K222 haplotype in the Dutch Toggenburger underlines the need for conservation of rare breeds in order to conserve genetic diversity rare or absent in other breeds.

Evaluation of genetic variability within PrP genotyped sheep of endangered Italian Altamurana breed

In the last few decades, there has been increased awareness of preservation and exploitation of endangered breeds for the maintenance of biodiversity, as well as the concern for diseases in sheep breeding.This study was carried out in native endangered Altamurana dairy sheep breed from Southern Italy. The Altamurana breed was considered as two populations (Alt-Cav and Alt-Cra-Zoe), based on presumed cross-breed and remarkable differences in the PrP genotypes frequencies. The genetic diversity between the two Altamurana populations (Alt-Cav and Alt-Cra-Zoe) was evaluated in comparison to three Italian dairy breeds through fourteen microsatellite markers. Both measures of genetic distance and the population structure analysis highlighted that the Alt-Cav and Alt-Cra-Zoe sheep have a particular genetic background. The estimated fixation index (FST) and the genetic Nei’s distances among the populations showed a higher level of genetic differentiation for Alt-Cav than Alt-Cra-Zoe. The Bayesian clustering analysis implemented in the STRUCTURE software showed clear and distinct clusters for the two Altamurana populations, confirming the hypothesis of Alt-Cav as a genetic group well differentiated from Alt-Cra-Zoe. Alt-Cav likely can be considered as belonging to the original strain of the Altamurana breed. This findings may be used to assist the programme for conservation and selection of scrapie resistance genotypes in endangered Altamurana sheep breed.

PRNP polymorphisms in four Italian sheep breeds

Prions are responsible for transmissible spongiform encephalopathies (TSEs), also called scrapie in sheep. Single nucleotide polymorphisms (SNPs) in PRNP have been shown to play a crucial role in terms of incubation period and/or susceptibility to scrapie. Considering codons 136, 154, 171, five main alleles, associated with different degrees of susceptibility to classical scrapie have been detected. Homozygous ARR sheep are resistant to scrapie infection, however this genotype is not so frequent in some breeds. VRQ and ARQ alleles on the other hand are associated with a high susceptibility to scrapie. Additional polymorphisms within the ARQ allele, have been associated with scrapie resistance. Therefore, it is important to assess the frequency of both main and minor allelic variants, in order to develop a breeding programme based on genetic selection that simultaneously, increases scrapie resistance, and maintains a high variability of PRNP. Allelic and genotypic PRNP frequencies, at the three main codons, were determined in four sheep breeds reared in central Italy (Appenninica, Bergamasca, Comisana and Sarda). Moreover homozygous ARQ sheep have been investigated to measure the frequencies of minor allelic variants. In this study the AT112RQ and ARQK176 protective allelic variants were detected. The AT112RQ allele was observed (ranging from 6.3% to 31.8%) in all the analysed breeds with the exception of Sarda sheep. The ARQK176 allele by contrast was observed only in Sarda breed (4.6%). Moreover five non synonymous polymorphisms were also found (Q101R, G127S, L141F, H143R, and H180Y).

Genetic and Pathological Follow-Up Study of Goats Experimentally and Naturally Exposed to a Sheep Scrapie Isolate.

Thirty-seven goats carrying different prion protein genotypes (PRNP) were orally infected with a classical scrapie brain homogenate from wild-type (ARQ/ARQ) sheep and then mated to obtain 2 additional generations of offspring, which were kept in the same environment and allowed to be naturally exposed to scrapie. Occurrence of clinical or subclinical scrapie was observed in the experimentally infected goats (F0) and in only one (F1b) of the naturally exposed offspring groups. In both groups (F0 and F1b), goats carrying the R154H, H154H, R211Q, and P168Q-P240P dimorphisms died of scrapie after a longer incubation period than wild-type, G37V, Q168Q-P240P, and S240P goats. In contrast, D145D and Q222K goats were resistant to infection. The immunobiochemical signature of the scrapie isolate and its pathological aspects observed in the sheep donors were substantially maintained over 2 goat generations, i.e., after experimental and natural transmission. This demonstrates that the prion protein gene sequence, which is shared by sheep and goats, is more powerful than any possible but unknown species-related factors in determining scrapie phenotypes. With regard to genetics, our study confirms that the K222 mutation protects goats even against ovine scrapie isolates, and for the first time, a possible association of D145 mutation with scrapie resistance is shown. In addition, it is possible that the sole diverse frequencies of these genetic variants might, at least in part, shape the prevalence of scrapie among naturally exposed progenies in affected herds. This study was aimed at investigating the genetic and pathological features characterizing sheep-to-goat transmission of scrapie. We show that in goats with different prion protein gene mutations, the K222 genetic variant is associated with scrapie resistance after natural and experimental exposure to ovine prion infectivity. In addition, we observed for the first time a protective effect of the D145 goat variant against scrapie. Importantly, our results demonstrate that the phenotypic characteristic of the wild-type sheep scrapie isolate is substantially preserved in goats carrying different susceptible PRNP gene variants, thus indicating that the prion protein gene sequence, which is shared by sheep and goats, plays a fundamental role in determining scrapie phenotypes.

Biodiversity and selection for scrapie resistance in goats: Genetic polymorphism in “Girgentana” breed in Sicily, Italy

Breeding for scrapie resistance in sheep, has being implemented in Europe as an efficient eradication policy. In goats, in contrast, the analysis on prion gene (PRNP) polymorphisms showed that some polymorphic variants can be protective for the disease but their prevalence in different breeds is still under investigation. Currently, in the presence of positive goats a complete cull of the herd is performed with a great economic loss and a serious threat of extinction for the endangered breeds.A polymorphism that seems related to caprine scrapie resistance is present at the codon 222 of the PRNP, with a lysine (K) instead of glutamine (Q) but this variant is not present in several widely diffused goat breeds in different countries. To evaluate the prevalence of the K222 variant in local goats we studied the “Girgentana”, an endangered autochthon dairy breed. The dairy products from the milk of this breed, with an excellent balance between fat and proteins are very typical and considered a “Slow Food” presidium.The results showed that K222 variant is common in Girgentana breed (18,7%) suggesting a rapid realization of a breeding plan for scrapie resistance based on K222 allele, in contrast to commercially widely used alpine breeds such as Saanen, that have a lower prevalence of this allele. The results suggest that genetic biodiversity might be a great resource for the selection of scrapie resistance, for the conservation of endangered breeds and their typical food productions.

Biodiversity and Genetic Polymorphisms Against Scrapie in Sopravissana Sheep Breed

Scrapie is a neurodegenerative disease affecting ovine and it is one of several transmissible spongiform encephalopathies (TSEs). Scrapie is recognized as two forms – classical and atypical. Susceptibility or resistance to classical scrapie is strongly regulated by the polymorphisms at codons 136, 154 and 171 of the PRNP gene. Genetic role in atypical scrapie has been described at codons 141 and 154, these ones are involved in the occurrence of the disease at different risk degrees. The aim of this study was to assess the allelic and genotypic frequencies in Sopravissana breed, an endangered autochthon breed of central Italy; in addition, the presence and the frequency of potential protective ARQ allele variants able to increase scrapie resistance by preserving a higher variability of PRNP were evaluated. Three alleles (ARQ, ARR and AHQ) and seven genotypes were observed based on codons 136, 154 and 171 with different frequencies. Moreover, the entire coding sequence of prion protein (PRNP) of the ARQ/ARQ was sequenced; two non-synonymous L141F (23.8%) and H143R (16.7%), also two synonymous polymorphisms (codons 231 and 237) were found. The results showed that the ARQ/ARQ sheep should be considered as a genetic class, which may potentially include animals with different scrapie susceptibility because of the presence of additional polymorphisms. This may allow the application of alternative strategies for breeding programmes in this endangered breed.

Assessment of inbreeding resulting from selection for scrapie resistance: a model for rare sheep breeds

Inbreeding in a small population of Chios sheep undergoing intense selection for the PrP gene was assessed 10 years after the beginning of a scrapie resistance selection programme. Inbreeding in...

Lack of relationship between Visna/maedi infection and scrapie resistance genetic markers

The relationship between Visna/maedi virus (VMV) antibody status and scrapie genetic resistance of 10,611 Rasa Aragonesa sheep from 17 flocks in Aragón (Spain) was investigated. The fifteen most common PRNP gene haplotypes and genotypes were identified and the genotypes were classified into the corresponding scrapie risk groups (groups 1 to 5). ARQ (93.3%) and ARR (31.8%) were the most common haplotypes and ARQ/ARQ (56%) and ARR/ARQ (25.6%) were the most common genotypes. The frequencies of scrapie risk groups 1, 2, 3, 4 and 5 were 3.3%, 27.3%, 63.5%, 1.2% and 4.8%, respectively. Overall Visna/maedi seroprevalence was 53% and flock seroprevalence ranged between 21-86%. A random effects logistic regression model indicated that sheep VMV serological status (outcome variable) was not associated with any particular scrapie risk group. Instead, VMV seropositivity progressively increased with age, was significantly greater in females compared to males and varied between flocks. The absence of a relationship between VMV infection and scrapie genotypes is important for VMV control and specifically for sheep participating in an ELISA-based Visna/maedi control program.

PRNP polymorphisms in Tunisian sheep breeds

In this study, genetic variation of ovine prion protein in Tunisian sheep breeds was analysed. Sequencing of the entire coding sequence of prion protein gene (PRNP) was performed in a total of 201 samples belonging to four breeds (Barbarin, Western Thin Tail, Sicilo Sarde and Black Thibar). Five haplotypes (ARQ, ARR, ARH, AHQ and VRQ) and 10 genotypes were observed based on codons 136, 154 and 171, with different frequencies among the investigated breeds. The ARQ, ARR and ARH haplotypes were present in all breeds, the VRQ haplotype was observed at low frequencies in Barbarin and Western Thin Tail breeds. The ARQ and ARR haplotypes were the most common with frequencies ranging from 33.4% to 47.8% and from 26.5% to 46.5% respectively, in the different breeds. Moreover, 12 additional non-synonymous (Q101R, M112T, G127S/V, M137T, L141F, H143R, N146S, R151G, Y172D, N176K and H180Y) as well as 2 synonymous polymorphisms at codons 231 and 237 were found in the PRNP gene. Among them, the R151G polymorphism has never been described in sheep. Moreover an insertion of an octarepeat in the ARQ haplotype has been observed. These results represent the first survey on PRNP variability in Tunisian sheep and may serve as basis for the development of breeding programme to increase scrapie resistance in national sheep breeds.

Perspectives of a scrapie resistance breeding scheme targeting Q211, S146 and K222 caprine PRNP alleles in Greek goats

The present study investigates the potential use of the scrapie-protective Q211 S146 and K222 caprine PRNP alleles as targets for selective breeding in Greek goats. Genotyping data from a high number of healthy goats with special emphasis on bucks, revealed high frequencies of these alleles, while the estimated probabilities of disease occurrence in animals carrying these alleles were low, suggesting that they can be used for selection. Greek goats represent one of the largest populations in Europe. Thus, the considerations presented here are an example of the expected effect of such a scheme on scrapie occurrence and on stakeholders.

Nanopore analysis reveals differences in structural stability of ovine PrPC proteins corresponding to scrapie susceptible (VRQ) and resistance (ARR) genotypes

Species, as well as individuals within species, have unique susceptibilities to prion infection that are likely based on sequence differences in cellular prion protein (PrPC). Species barriers to transmission also reflect PrPC sequence differences. Defining the structure-activity relationship of PrPC/PrPSc with respect to infectivity/susceptibility will benefit disease understanding and assessment of transmission risks. Here, nanopore analysis is employed to investigate genotypes of sheep PrPC corresponding to differential susceptibilities to scrapie infection. Under non-denaturing conditions scrapie resistant (ARR) and susceptible (VRQ) genotypes display similar, type I (bumping) predominant event profiles, suggesting a conserved folding pattern. Under increasingly denaturing conditions both proteins shift to type II (intercalation/translocation) events but with different sensitivities to unfolding. Specifically, when pre-incubated in 2M Gdn-HCl, the VRQ variant had more of type II events as compared with the ARR protein, suggesting a more flexible unfolding pattern. Addition of PrPSc-specific polyclonal antibody (YML) to the ARR variant, pre-incubated in 2M Gdn-HCl, reduced the number of type II events with no clear intercalation/translocation peak, whereas for VRQ, type II events above blockades of 90 pA bound YML. A second PrPSc-specific antibody (SN6b) to a different cryptic epitope reduced type II events for VRQ but not the ARR variant. Collectively, the event patterns associated with sequential denaturation, as well as interactions with PrPSc-specific antibodies, support unique patterns and/or propensities of misfolding between the genotypes. Overall, nanopore analysis identifies intermediate conformations that occur during the unfolding pathways of ARR and VRQ genotypes and may help to understand the correlation of structural properties that induce protein misfolding.

Effect of selection for scrapie resistance on genetic diversity in a rare and locally adapted sheep breed: The case of Sambucana

In Italy, since 2005 a breeding plan to increase scrapie resistance has been adopted. The impact of this selection on genetic diversity was assessed on Sambucana, an autochtonous sheep breed reared in southern Piedmont, by analysing the evolution of allele frequencies at different levels: PRNP (prion protein) gene, microsatellite loci on OAR13 (where PRNR maps), and microsatellite loci on other chromosomes, not subjected to selection for scrapie resistance. A total of 147 young rams, 80 born in 2004 and 67 in 2008–2009 were analysed. Evidence of diversity loss was observed for PRNP gene as a consequence of the directional selection. Diversity was affected in the immediate vicinity of PRNP but the effect on more distant loci on the same chromosome was trivial. With regard to neutral markers, lack of heterozygosis with no changeover of allele frequencies was observed suggesting an increase of inbreeding. Mating policies would be sufficient to solve these problems. A selection scheme based on genotyping rams and eliminating carriers of both susceptible and high susceptible alleles is the best way to improve natural resistance to scrapie with low costs and minimal problems in the current conservation programmes targeting rare breeds.

Short communication: Analysis of association between the prion protein (PRNP) locus and milk traits in Latxa dairy sheep

The purpose of this study was to analyze associations between polymorphisms in the PRNP gene and ewe milk traits. A total of 242,565 lactations of the Latxa breed were used. Milk, fat and protein yields, and fat and protein content from black-faced Latxa from Spanish Basque Country, black-faced Latxa from Navarra, and blond-faced Latxa were collected. To evaluate evidence of association, the different traits were analyzed using an animal model, where the PRNP genotype effect was included or not as a random effect. Adding the PRNP effect to the model improved the fitting for milk yield in black-faced Latxa from Spanish Basque Country and in blond-faced Latxa, for fat yield in black-faced Latxa from Navarra, and for protein yield in blond-faced Latxa. However, the proportion of the phenotypic variance explained by the PRNP effect for milk yield (1.0×10−3), fat yield (3.6×10−3) and protein yield (9.4×10−4) were near zero. The PRNP locus accounts for about 0.5, 1.5, and 0.4% of total genetic (PRNP and polygenic) variance in milk, fat, and protein yield. These values indicated that the PRNP effect is not relevant regarding genetic additive contribution. For breeding purposes, it is unlikely that selection for scrapie resistance will have an effect on the milk traits studied in the Latxa breed.

Genetic Variability of PRNP in Chinese Indigenous Goats

Polymorphism of the prion protein gene (PRNP) is usually associated with scrapie susceptibility or resistance. To determine the variability of PRNP in Chinese indigenous goat breeds, we isolated genomic DNA from goat blood and amplified and sequenced the coding region of the gene. We identified 10 polymorphic sites that gave rise to 28 haplotypes. Clear frequency differences were found between northern and southern breeds and confirmed by genetic distance analysis, except for the Tangshan dairy goat. Phylogeographic analysis supported the idea that northern and southern breeds might be considered separate clusters, except for the Tangshan dairy goat. The finding of significant differences in allele distribution in northern and southern goats, especially if involved in modulating resistance/susceptibility, needs to be carefully considered for the feasibility of selection plans for resistance to scrapie.