Breeding for scrapie resistance in the Hungarian sheep population

The first results of the Hungarian sheep prion protein (PrP) genotyping programme are discussed in this paper. To obtain initial genotype frequency data 10 commercial (Hungarian Merino, German Mutton Merino, Merino Landschaf, German Blackheaded, Suffolk, Texel, Ile de France, Charollais, Lacaune, British Milksheep) and 4 indigenous (Gyimes Racka, Hortobágy Racka, Tsigaja, Cikta) breeds were sampled in 2003 and 2004, and the PrP genotypes were determined by microsequencing analysis with capillary electrophoresis. In all commercial breeds, a higher number of sheep were genotyped in 2005 (3648) and in 2006 (3834) within the breeding programme to increase scrapie resistance, and the estimated frequency data were compared to the initial figures to evaluate the efficiency of selection. The new developments arising from the identification of the so-called 'atypical' scrapie cases are also discussed.     

PrP genotype frequencies in German breeding sheep and the potential to breed for resistance to scrapie

Genetic susceptibility to scrapie is associated with polymorphisms in three different codons of the ovine prion protein (PrP) gene (136, 154, 171). Studies of PrP genotypes linked to scrapie have revealed the resistance of homozygous PrPARR/PrPARR animals and the high risk of PrPVRQ/PrPVRQ and PrPvRQ/PrPARQ animals in scrapie-affected flocks. The selection of PrPARR/PrPARR genotypes may therefore provide a strategy for controlling clinical scrapie. The genotypes of 1361 German breeding sheep from 15 different breeds in northern Germany were determined. Apart from the wildtype allele PrPARQ, at least four mutually exclusive allelic variants were found. The greatest variability within the PrP gene was encountered in texel sheep, in which 14 PrP genotypes were found. In the important meat breeds, Suffolk, German whiteheaded mutton and German blackheaded mutton, the PrPARR allele was predominant, and in these breeds the breeding of scrapie-resistant pedigree flocks within four generations seems to be a feasible option. In the texel sheep, the German merino, the German milk and the German land sheep breeds, the frequency of the PrPARR allele was much lower, and in several breeds no homozygous rams were available for breeding purposes. In these breeds the breeding strategy would depend on the number of heterozygous rams available, but resistant pedigree flocks could be achieved within nine generations.     

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.     

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

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

Genetic diversity loss due to selection for scrapie resistance in the rare Spanish Xalda sheep breed

The effect of selection for scrapie resistance on genetic variability in the rare Xalda sheep breed was studied. Pedigree information comprised 1851 animals (1444 alive) at the moment of sampling. A total of 304 reproductive (or selected for reproduction) Xalda individuals were sampled and genotyped for 14 microsatellites. Genetic variability was assessed using: gene diversity (1 − average kinship, GD), mean average relatedness (AR) and self-coancestry (c_i) at the genealogical level; and expected heterozygosity (H_e), molecular mean kinship (Mk), molecular self-coancestry (s_i) and rarefacted average number of alleles per locus (A) at the molecular level. Two breeding strategies were evaluated: a) use of only young rams with genotype ARR/ARR and young ewes with low to moderate risk (risk groups R1 to R3); b) breeding without selection for PrP genotypes. The major cause of losses of genetic variability in the Xalda breed is the drift that occurs when a new group of reproductive individuals is selected. The loss of genetic variability is small in females compared to the males, where it is considerable. However, losses at the molecular level for young females with respect to adult females were above 5%. Young male individuals also retained most of the genetic variability assessed in adult rams. Selection against susceptibility to scrapie produced additional losses of more than 2% for H_e and A when rYF individuals are considered. As regards males, the situation becomes critical because of the scant number of available ARR/ARR young rams. The consequences for the management of the Xalda breed are discussed.     

Quantifying diversity losses due to selection for scrapie resistance in three endangered Spanish sheep breeds using microsatellite information

The effect of selection for scrapie resistance on genetic variability in three endangered Spanish sheep breeds (Colmenareña, Mallorquina and Rubia de El Molar) was studied using two different criteria for quantifying contributions to genetic variability: (a) molecular coancestry or genetic identity; and (b) average number of alleles per locus or allelic richness. A total of 236 (81 Colmenareña, 76 Mallorquina and 79 Rubia de El Molar) individuals were genotyped for the PrP gene and for 22 microsatellite markers. The analyses assumed a selective policy aimed at the elimination of the VRQ allele and the reduction of the frequency of the ARQ/ARQ genotype. These goals are approached by rejecting for breeding those individuals with the highest susceptibility for scrapie (risk groups R4 and R5) in a genetic scenario with no previous selection programmes considering the PrP gene polymorphism carried out. When all the individuals classified into risk groups R4 and R5 were removed from the dataset, the total molecular coancestry slightly increased in the Colmenareña breed illustrating that the carriers of undesirable PrP genotypes are not essential to maintain its overall gene diversity. When the allelic richness was considered, the removal of the R4 and R5 individuals gave high losses in the Rubia de El Molar breed. The analyses carried out considering the sex of the individuals informed that most increases in genetic identity in the Colmenareña breed resulted from the removal of the R4 and R5 males while in the Mallorquina breed resulted from the removal of the undesirable females. Losses of diversity in the Rubia de El Molar breed were basically independent of the sex of the individuals due to the balanced contributions to diversity of both sexes. As a general recommendation, not all the individuals of undesirable risk groups should be rejected for reproduction at the same time to avoid irretrievable loses of genetic diversity but according to the sex of the individuals.     

Merino sheep breed's genetic resistance to scrapie: genetic structure and comparison of five eradication strategies

Three thousand one hundred and ninety-three Merino-breeding males were genotyped for Scrapie-resistance gene. They showed a high frequency of ARQ allele while VRQ allele frequency, linked to the highest susceptibility, was close to 0. Alleles linked to Scrapie-susceptibility frequencies have been studied in different Merino flocks. Most of the variability is intraflock, not interflock. In addition, there is an excess of heterozygotes due to crossbreeding or Wahlund effect.

Five control and prevention strategies were studied: first genotyping males and females and eliminating VRQ carriers and ARQ/ARQ rams; second genotyping males and females and eliminating VRQ carriers; third genotyping males and eliminating VRQ carriers and ARQ/ARQ rams; fourth genotyping males and using only as breeders ARR/ARR, ARR/AHQ, AHQ/AHQ and ARQ/ARQ rams; fifth genotyping males and using only as breeders ARR/ARR, ARR/AHQ and AHQ/AHQ rams. They were simulated from allelic and genotypic frequencies to decide which animals had to be genotyped and which animals had to be eliminated due to their genotypes and risk levels. The third option, to genotype rams and to eliminate ARQ/ARQ and VRQ carriers, would be the best strategy to improve the resistance and would cause minimal cost and loss of animals.     

Breeding for resistance to mastitis in United Kingdom sheep, a review and economic appraisal

Mastitis is a problem in the sheep industry, and its incidence varies widely with how it is recorded, the breed of sheep and the farm. Virtually all the published information about the genetics of mastitis refers to dairy breeds of cattle and sheep, and there is little information for meat sheep breeds. Many dairy breeding programmes worldwide use the somatic cell count (scc) in milk as an indicator of resistance to clinical and subclinical mastitis, but it is difficult to measure in meat sheep breeds. Molecular genetic technologies may therefore be a more practical way to assess susceptibility to mastitis. This paper reviews the genetics of mastitis and considers the opportunities for breeding for resistance to mastitis, with particular reference to sheep. In addition, to investigate the potential economic effects of mastitis in a purebred sheep population, a computer model of flock dynamics was developed. By making a modest set of assumptions about the key farm parameters that influence lowland sheep production, the model showed that breeding for resistance (or other control methods), if it could reduce the risk of contracting mastitis by 10 per cent, would be worth pounds 8.40 per ewe, equivalent annually to pounds 2.7 million for the purebred sector of the Texel breed alone.     

Selection of Swaledale sheep of reduced susceptibility to experimental scrapie

A flock of 294 Swaledale sheep was injected with brain tissue from different sources so as to include strains of natural scrapie currently affecting the Swaledale breed. Discounting intercurrent deaths, 85 per cent of the flock developed clinical scrapie within an observation period of 2557 days (seven years). Scrapie cases fell into distinct early and late groups, occurring at (mean +/- sem) 287 +/- 7 days and 1207 +/- 38 days after injection, respectively. Breeding groups were formed from the survivors after the early cases had occurred and successive lamb crops were produced. Two groups of progeny and two groups of controls were injected with scrapie. The incidence of scrapie in the progeny (27 per cent) was much lower than in the controls (64 per cent). The incidence in the progeny was also assessed in relation to the response of their parents: it was high (about 40 per cent) when one or both parents developed late scrapie but otherwise very low (about 5 per cent). Hence the progeny of surviving parents had a greatly reduced susceptibility to the strains of scrapie used. Seventy-two of these selected progeny formed a nucleus flock which is being bred at Redesdale. The next stage of the experiment is to assess the effect of selected rams in recorded flocks with a high incidence of natural scrapie.     

Selection for scrapie resistance and simultaneous restriction of inbreeding in the rare sheep breed "Mergellander"

Scrapie is a fatal infectious neurodegenerative disease for which susceptibility is associated with polymorphisms in the ovine prion protein (PrP) gene. Scrapie-eradication programmes are based on eliminating the susceptible VRQ allele and/or breeding for the resistant ARR allele. In rare breeds or breeds with a low frequency of the ARR allele this can lead to unacceptably high inbreeding rates with associated increased risk of genetic defects and inbreeding depression. The conservation status of populations with inbreeding rates (DeltaF) above 1% is considered critical. In the Dutch rare sheep breed the Mergellander animals carrying ARR alleles are closely related to one another, and could reach 1.53% when only ARR/ARR animals are used as parents. Inbreeding rates can be reduced by selecting the set of parents according to their average co-ancestry. We minimised inbreeding rates by calculating the optimal contribution of each ram and selection of ewes. This resulted in inbreeding rates of -0.17% with exclusive use of homozygous ARR rams, and -0.38% if use of heterozygous rams was allowed as well. Thus sophisticated breeding programs can prevent unacceptably high inbreeding rates when breeding for scrapie resistance.     

Breeding objectives for Targhee sheep

Breeding objectives were developed for Targhee sheep under rangeland production conditions. Traits considered were those for which EPD were available from the US National Sheep Improvement Program and included direct and maternal effects on 120-d weaning weight (WW and MM, respectively); yearling weight (YW); yearling fleece weight, fiber diameter, and staple length; and percent lamb crop (PLC), measured as the number of lambs born per 100 ewes lambing. A bioeconomic model was used to predict the effects of a change of 1 additive SD in EPD for each trait, holding all other traits constant at their mean, on animal performance, feed requirements, feed costs, and economic returns. Resulting economic weightings were then used to derive selection indexes. Indexes were derived separately for 3 prolificacy levels (1.41, 1.55, and 1.70 lambs/ewe lambing), 2 triplet survival levels (50 and 67%), 2 lamb pricing policies (with or without discounting of prices for heavy feeder lambs), and 3 forage cost scenarios (renting pasture, purchasing hay, or reducing flock size to accommodate increased nutrient requirements for production). Increasing PLC generally had the largest impact on profitability, although an increase in WW was equally important, with low feed costs and no discounting of prices for heavy feeder lambs. Increases in PLC were recommended at all 3 prolificacy levels, but with low triplet survival the value of increasing PLC eventually declined as the mean litter size increased to approximately 2.15 lambs/ewe lambing and above. Increasing YW (independent of WW) increased ewe maintenance costs and reduced profitability. Predicted changes in breeding values for WW and YW under index selection varied with lamb pricing policy and feed costs. With low feed costs or no discounts for heavy lambs, YW increased at a modest rate in association with increasing WW, but with high feed costs or discounting of heavy lambs, genetic trends in WW were reduced by approximately 50% to constrain increases in YW. Changes in EPD for MM or fleece traits generally had smaller effects on profitability than changes in PLC, WW, and YW. Two indexes designed to address current rangeland production conditions (low forage costs and discounting of heavy feeder lambs) or more intensive and integrated production with retained ownership and value-based marketing of lambs (higher forage costs and no discounting of heavy lambs) were anticipated to meet the needs of most Targhee producers.     

Effects of different selection strategies on breeding towards scrapie resistance in the German Grey-horned Heath population

Scrapie resistance is related to polymorphisms of the prion protein (PrP) gene. The homozygous ARR/ARR genotype of the prion protein gene is associated with high resistance against conventional scrapie infections. But this can lead to a distinct loss of genetic variation in breeds with a small population size or a low frequency of the ARR allele. For these populations an optimal breeding scheme for breeding towards scrapie resistance and an arbitrarily chosen production trait with conservation of the genetic diversity is not known yet. A simulation programme was developed, in which the structures of local populations could be used as input parameters, and the development of the ARR allele frequency, of the inbreeding coefficient, of a production trait and the genetic drift were computed in dependence of different selection schemes for scrapie resistance. An optimal strategy for the German Grey Heath population from Lower Saxony should be found. An optimal strategy in a breeding programme for ARR homozygosis should also maintain the genetic diversity present in a breed. This could be achieved when initially ARR heterozygous sheep were bred until a certain threshold value of the ARR allele was reached. The speed and costs of the breeding progress towards fixation of the ARR allele should then be optimized with regard to the aims of the breeders and breeding organisations, whereas a faster breeding towards scrapie resistance goes along with higher genotyping costs and vice versa.     

Susceptibility of transgenic mice expressing chimeric sheep,bovine and human PrP genes to sheep scrapie

The use of Transgenic (Tg) mice expressing chimeric sheep/mouse (Sh/Mo) prion protein (PrP) and chimeric bovine/mouse (Bo/Mo) PrP genes was evaluated as a sheep scrapie model. We also investigated the potential for the transmission of sheep scrapie to a human/mouse (Hu/Mo) PrP Tg mouse line. The Sh/Mo PrP and Bo/Mo PrP Tg Prnp(+/+) or Prnp(0/0) mouse lines were inoculated intracerebrally with brain homogenates from three sheep with natural scrapie (KU, Y5 or S2). Incubation periods were slightly shorter in Sh/Mo PrP Tg Prnp(+/+), than in non-Tg mice inoculated with KU brain homogenate. In contrast, the incubation period was significantly prolonged (p<0.05) in Bo/Mo PrP Tg Prnp(+/+) mice inoculated with KU brain homogenate. The incubation period was significantly longer in all Tg Prnp(+/+) and Prnp(0/0), than in non-Tg mice (p<0.01) inoculated withY5 brain homogenate. None of the Tg Prnp(0/0) mice inoculated with S2 brain homogenate developed clinical signs and PrP(Sc) was undetectable in their brains. These results suggested that expression of the Sh/Mo PrP or Bo/Mo PrP transgenes does not confer susceptibility to sheep prions upon mice, and thus none of the Tg mouse lines could be a suitable model of sheep scrapie. Hu/Mo PrP Tg Prnp(0/0) mice inoculated with natural and experimental scrapie or mouse prions did not develop clinical signs of scrapie and PrP(Sc) was undetectable. These results suggested that neither sheep nor mouse strains of scrapie are highly transmissible to humans.