Breeding programmes for TSE resistance in British sheep. I. Assessing the impact on prion protein (PrP) genotype frequencies

From April 2005, member states of the European Union were required to implement a compulsory breeding programme for resistance to transmissible spongiform encephalopathies (TSEs) in sheep as part of measures to eradicate TSEs from national flocks. In this paper, we assessed the impact of four different breeding strategies on prion protein (PrP) genotype frequencies using a mathematical model which describes in detail gene flow in the British sheep flock. These strategies ranged from the minimum requirements laid down in by EU legislation to compulsory implementation of the current National Scrapie Plan for Great Britain (NSP) ram genotyping scheme. All four strategies were predicted to substantially reduce the frequency of the VRQ allele, which is associated with the highest risk of scrapie, although schemes with more stringent requirements produced a larger reduction. However, there were marked differences in the impact of the strategies on the frequency of other PrP alleles. In particular, restrictions beyond those required by EU legislation were necessary to change the frequency of other PrP alleles substantially. Consequently, a breeding programme which aims to reduce the risk to human health by reducing the frequency of the ARQ allele (associated with the highest risk of BSE in sheep) must place restrictions on ARQ-bearing animals. Similarly, a programme which seeks to increase the frequency of the ARR allele (associated with the lowest risk of TSE) must favour ARR-bearing animals.     

Eradication of footrot of lesser clinical severity (intermediate footrot)

OBJECTIVE: To determine if intermediate footrot (IFR) can be eradicated from a flock of sheep by inspection and culling of cases during a non-transmission period and if prior antibiotic treatment or vaccination increases the likelihood of eradication. PROCEDURE: A replicated field experiment that compared the three eradication strategies was followed by an observational study of the best of these applied in a commercial flock of 3000 sheep. RESULTS: In the replicated experiment, IFR was eradicated either by inspection and culling alone, or when combined with vaccination. Eradication failed when the sheep were treated with parenteral antibiotics before inspection and culling during the non-transmission period. In the whole-flock program, eradication by repeated inspection and culling of footrot cases during the non-transmission period was successful and the flock remained free of infection 3 years later. CONCLUSIONS: IFR can be eradicated by inspection and culling but latent infections, which may persist undetected for at least 34 weeks, require surveillance inspections to be repeated during the non-transmission phase of the program. The use of parenteral antibiotics as an aid to the eradication of IFR is contraindicated.     

TSE eradication in small ruminants--quo vadis?

The term of 'TSE infections in small ruminants' summarises BSE as well as classical and the recently discovered atypical scrapie infections in sheep and goats.There are fundamental differences between the TSE infections in small and large ruminants. Other than in bovines the TSE pathogenesis in small ruminants implies that various peripheral tissues become infectious long before the onset of clinical symptoms. At least in sheep, classical scrapie is efficiently transmitted horizontally within affected flocks. On the other hand, BSE poses a distinctly higher zoonotic risk than scrapie. Therefore, regulatory measures for the protection of animals and humans from a BSE infection must be substantially different for large and small ruminants. While culling of the birth and feeding cohort of a BSE affected cattle is considered to be effective to prevent any further BSE cases in the affected herd, an effective BSE and classical scrapie eradication programme in small ruminants requires a much more stringent eradication strategy and the rendering of all susceptible animals. The situation became even more complicated when atypical scrapie cases with divergent transmission and pathogenesis characteristics and with a novel biochemical phenotype of the infectious agent came into play. The discovery of these atypical scrapie cases has initiated a discussion about the suitability of the current TSE eradication measures in sheep (which are selective breeding and genotype based culling), in particular when such cases were also found in sheep carrying the believed scrapie resistant genotypes.     

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.     

Active surveillance for scrapie in the Netherlands: effect of a breeding programme on the prevalence of scrapie in sheep (2002-2010)?

The susceptibility of sheep to scrapie is modulated by the prion protein (PrP) genotype of the animal. An ambitious voluntary scrapie control programme was started in the Netherlands in 1998, based on selection of rams with theARR/ARR genotype for breeding. This programme was followed by an obligatory programme in 2004; the programme has been voluntary since 2007. We monitored the prevalence of PrP genotype frequencies and the prevalence of scrapie in the Dutch sheep population between 2002 and June 2010. Results showed that selection for scrapie-resistant sheep resulted in an increase in the ARR allele frequency in the Dutch national flock from 37.5% in 2005 to 61.4% in 2009. Moreover, surveillance data showed that there was a significant decrease in the prevalence of scrapie a few years after the start of the obligatory breeding programme, from more than 0.2% in 2004 to 0.015% in 2009. This decrease is a consequence of the increased number of scrapie-resistant sheep in the Dutch sheep population. To date, the results and the models based on the data show that the selective breeding programme should be continued for several years in order to successfully eradicate scrapie. It will be important to monitor the PrP frequency and scrapie prevalence in the Dutch sheep population in the coming years.     

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.     

Assessing the efficacy of a ram-genotyping programme to reduce susceptibility to scrapie in Great Britain

Susceptibility to clinical scrapie is associated with polymorphisms in the PrP gene. The 'ARR' allele of this gene reduces susceptibility to clinical disease caused by all known strains of the transmissible spongiform encephalopathy (TSE) agent. The British government proposes to use a ram-genotyping scheme to breed genetic resistance to clinical scrapie into the national sheep population. We considered how best to target limited genotyping resources to achieve the maximum rate of genotype evolution. We created a metapopulation model of the British sheep industry, which includes the major pure-breeds of sheep and the cross-breeds produced by crossing these pure-bred animals. The main criterion for assessing the efficacy of different strategies was the time taken to increase the prevalence of the ARR allele in the slaughter-lamb population. Our model predicted that the most-effective strategy would be to target genotyping to those rams used for pure-breeding (i.e. mated with the same breed of ewe). This strategy was compared to two further strategies, in which the proportion of rams genotyped in each breed depended on the prevalence of the ARR/ARR genotype in that breed. A policy in which the proportion of animals genotyped is reduced as the ARR prevalence in that breed increases is efficient. The most-effective policy was targeting the hill sector in the early years and gradually switching to genotyping more terminal-sire and longwool rams as the resistance of the hill sector increases.     

Progress and limits of PrP gene selection policy

Classical scrapie has proved to be a notoriously difficult disease to control due to a poor understanding of its natural history. The recognition of disease risk linkage to PrP genotype has offered the prospect of a disease control strategy, viz. genotyping and selective breeding, novel to veterinary medicine when first considered in the 1990s. The UK Spongiform Encephalopathy Advisory Committee recommended the exploitation of this approach in a voluntary, national programme to control classical scrapie and protect the public against food-borne exposure to bovine spongiform encephalopathy, should the national flock have been exposed via contaminated feed. The National Scrapie Plan for Great Britain was launched in 2001 and uptake has been widespread throughout the purebreeding sector of the sheep industry, with membership peaking at over 12 000 flocks in 2006. A total of 700 000 rams from 90 breeds have been genotyped. A comparison of ram lambs born in 2002 with those in 2006 shows evident changes in PrP genotype frequencies which are predicted to be associated with a reduction in disease risk. Various concerns have been raised regarding possible unintended consequences of widespread selection on PrP genotype, including impacts on other performance traits and possible effects on inbreeding and genetic diversity. To date, these concerns appear to be unfounded, as no consistent associations have been found with performance traits, nor are there likely to be any detectable impacts on inbreeding in mainstream breeds. Currently, semen banks have been implemented in Great Britain to store samples from animals of all common PrP genotypes, should these genotypes be required in the future. Various strategies to minimise future disease risks are discussed in the paper.     

A questionnaire survey of the prevalence of scrapie in sheep in Britain

An anonymous, self-administered questionnaire has been used in two independent surveys to try to determine the prevalence of scrapie in the national sheep flock. The disease was recorded in 35 counties in England and Wales. About a third (26.5 and 37.3 percent) of respondents owning 100 or more sheep indicated that they had seen sheep with scrapie in their flocks. The incidences of clinical cases recorded in affected flocks in the two surveys were 0.5 and 1.1 cases/100 ewes/year. At present there is no control over the disposal of these animals. If as has been suggested, an increase in the prevalence of scrapie was a contributory factor in the emergence of bovine spongiform encephalopathy, it would seem logical that measures should be introduced to monitor the prevalence and incidence of scrapie and to control the disposal of clinical cases.     

Ovine scrapie. New tools for control of an old disease.

Ovine scrapie was described nearly 300 years ago and is endemic in many parts of the world. The recent emergence of a related bovine disease in the United Kingdom and Europe, with probable transmission to humans, has lent urgency to scrapie surveillance and control programs. The biology, genetics, diagnosis, and proposed routes of transmission can be understood in the context of the presumed causative agent, the prion protein. An integrated program of management and husbandry to reduce introduction and spread of the disease within a flock, diagnosis of preclinically infected sheep in both live animal and postmortem settings, and identification of breeding stock of low risk of scrapie are reviewed as the basis for scrapie eradication programs.     

Descriptive epidemiology of scrapie in Great Britain: results of a postal survey

In 1998, a questionnaire was sent to 11,554 British sheep farmers to determine how many believed that scrapie cases had occurred in their flock; 61.4 per cent of them responded anonymously. The results indicated that 14.9 per cent of farmers with more than 30 breeding ewes thought that they had ever experienced scrapie in their flock and 2.7 per cent thought that they had had cases in the past 12 months. A comparison of these results with the number of farmers reporting suspect scrapie cases to MAFF, in accordance with the statutory requirement, suggests that only 13 per cent of farmers who suspect that they may have cases of scrapie are currently reporting them. Scrapie occurred in all regions of the country but there was an apparent regional variation. Larger farms and those with purebred sheep appeared to be at greater risk of having cases. Other differences between affected and unaffected farms included lambing practices and sheep purchasing policy. On the majority of farms the first case occurred in a purchased animal. The survey also revealed a need for the provision of further information about scrapie to farmers.     

Preventing the spread of maedi-visna in sheep through a voluntary control programme in Finland

The sheep disease maedi-visna (MV) was introduced into Finland in 1981 and had spread to eight flocks in the southwestern part of the country when first detected in a survey in 1994. Six more seropositive flocks were subsequently traced, bringing the total to 14. MV has a notifiable disease status in Finland that provides for official restrictive measures to which all infected herds are subject. These measures are withdrawn once the seropositive animals and their progeny are culled and the flock has showed negative signs in the test done twice, or after total culling. A voluntary control programme was initiated in January 1995 to extend official control efforts. The programme furnishes a guideline for culling, restrictions on contacts, and a timetable for testing the flock to attain MV-free status. Seven flocks of the 14 were slaughtered either immediately or after a period under restrictive measures. One flock finished sheep production after four years under restrictive measures. Selective culling and repeated testing was attempted with the other six flocks, three of which attained MV-free status. One flock finished sheep production after two years in the control programme, the other two dropped out of the programme when the restrictive measures were withdrawn. It was concluded that the control programme was salient in eradicating MV from Finland and that serological monitoring of the situation must be continuous.     

A survey of more than 11 years of neurologic diseases of ruminants with special reference to transmissible spongiform encephalopathies (TSEs) in Greece

The first cases of scrapie were detected in Greece in a flock of sheep in October 1986. All the animals of the affected flock and all sheep in two flocks that were in contact were killed and buried. A systematic investigation of all available cases with signs indicating a neurological disease started in sheep and goats in late 1986, as well as in cattle in 1989. The investigation was based on clinical examination, necropsy or macroscopical examination of the brain and viscera, and histological examination of the brain in all animals except those with coenurosis. Histological examinations of specimens from the spinal cord and other tissues, and if considered necessary bacteriological, toxicological and serological examinations were also carried out. In October 1997, scrapie was diagnosed in sheep of a second flock (a mixed flock of sheep and goats), grazing in a pasture close to the place where scrapie was initially detected. All animals of the second flock were also killed and buried. Diagnosis in the first flock was based on clinical signs and histological lesions, and in the second immunoblotting was also used. Distinctive lesions of scrapie were found in the brain and/or the spinal cord of eight sheep with clinical signs from the two flocks. The lesions were revealed in the brain stem and/or in the cervical spinal cord, and tended to be symmetrical. In one sheep, severe lesions in the cortex of cerebral hemispheres and of the cerebellum were also found. In the brain of two sheep from the second flock the pathological isoform of PrP protein was detected. Despite the eradication scheme applied, scrapie in sheep reappeared after 11 years in a place close to where it occurred initially. This may indicate that the effectiveness of the eradication scheme implemented was not adequate and additional approaches may be needed.     

Derivation of a scrapie-free sheep flock from the progeny of a flock affected by scrapie

The Cheviot flock at the Institute for Animal Health's Neuropathogenesis Unit (npu) has endemic scrapie, which affects primarily vrq/vrq sheep and at high frequency. A new flock with a full range of PrP genotypes, including the highly susceptible vrq/vrq, has been produced on a separate site, from animals in the npu breeding flock, and it remains scrapie-free after eight years. In contrast, in a parallel flock at the npu farm, scrapie has reappeared after five years, although the animals were kept in separate accommodation from the scrapie-affected sheep. During this time the npu breeding flock continued to have scrapie cases. Although it is known that highly susceptible sheep can remain free of infection in a clean environment, this is the first report of the infection being removed successfully from the bloodlines of scrapie-affected sheep. The results confirm that scrapie is not a genetic disease dependent only on the PrP gene sequence, but requires both genetic susceptibility and an infectious agent.     

A paired case-control study of risk factors for scrapie in Irish sheep flocks

We did a case–control study of the association of several animal, flock and management factors with scrapie in Irish sheep flocks. The characteristics of 61 sheep flocks with at least one laboratory-confirmed case of scrapie (1990–1998) were compared to 61 flocks with no history of scrapie and matched by geographical location and attending veterinary surgeon. The 61 scrapie-affected flocks were from the database of known scrapie flocks in the Republic of Ireland at the start of the study. In conditional multiple logistic regression, factors associated with increased odds of scrapie in a sheep flock were (i) larger breeding-flock size, (ii) purchasing replacement sheep through the market, (iii) spreading sheep compost on the land and (iv) disposing of the placenta in the compost. Factors associated with decreased odds of scrapie were (i) using cattle slurry on the land and (ii) feeding proprietary concentrates to lambs.     

Determining the relationship between restorative potential and size of a gene bank to alleviate the risks inherent in a scrapie eradication breeding programme

Breeding plans are being adopted in many countries to eradicate scrapie from the sheep populations. The European plans are based on selection for increased frequency of the resistant ARR haplotype and removal of the most scrapie susceptible haplotype (VRQ). This approach of a genotype eradication breeding programme incurs a number of risks including those associated with novel spongiform encephalopathy causing agents that are as yet unknown, but may emerge in the future and the possible loss of favourable attributes of a breed either through linkage or pleiotropic effects when selecting on a specific haplotype. Semen gene banks have been proposed to provide the potential for future reintroduction of removed haplotypes. Computer simulations were run for hill, crossing and terminal breed types to investigate the restorative potential of a gene bank required for the reintroduction of an alternative haplotype in the future. Differences existed in the restorative potential between the different breed types considered. This was largely dictated by variable levels of fertility, litter size, survival and mating ratio. It was shown that terminal breeds would require two and a half times as many semen straws as hill breeds to produce the same frequency of ewes homozygous for the desired haplotype in the population after a ten year reintroduction period.     

Investigation and control of mastitis outbreaks caused by Pseudomonas aeruginosa in a sheep flock and a goat herd

Outbreaks of mastitis in a sheep flock and a goat herd caused by Pseudomonas aeruginosa, and their control are described.The morbidity rates of mastitis in a sheep flock and in a goat herd were 18.7 and 18.2 per cent, respectively. The lethality and involuntary culling rates of the animals with sub-clinical mastitis correspond to the morbidity rate.The highest morbidity rates were noted in yearlings--22.5 and 30 per cent, as well as in animals pre- and post partum (7 and 20 per cent, in sheep and goats, respectively). The morbidity periods in the flock and in the herd were 20 and 27 months, respectively. A strict-hygiene milking programme and maintenance of the milking devices was introduced. In addition, prevention of crowding, provision of dry bedding and thorough cleaning of water troughs have also been instituted. Ruthless culling of all infected and carrier animals was an essential part of the control program against Pseudomonas aeruginosa mastitis.     

Caseous lymphadenitis in small ruminants.

Caseous lymphadenitis is a contagious bacterial disease that affects sheep and goats. It is characterized by abscess formation in the skin, internal and external lymph nodes, and internal organs. The causative agent is Corynebacterium pseudotuberculosis. The disease can become endemic in a herd or flock and is difficult to eradicate by virtue of its poor response to therapeutics, its ability to persist in the environment, and the limitations in detecting subclinically affected animals. The disease causes significant economic impact on the small ruminant industry through decreased meat yield, damaged wool and leather, decreased reproductive efficiency, culling of affected animals, and mortality from the internal environment.     

Breeding German sheep for resistance to scrapie

Susceptibility to clinical scrapie is associated with polymorphisms in the prion protein (PrP) gene. The ARR allele reduces susceptibility to clinical disease caused by all known strains of the transmissible spongiform encephalopathy (TSE) agents. For the economically important German breeds of sheep the PrP allele frequencies are well known, but this paper presents representative genotyping results for 1526 sheep from two smaller milk sheep breeds and 2446 sheep from 14 mostly indigenous land sheep breeds. The ARR allele was detected in each breed but the breed-specific ARR frequencies varied between 1 and 63 per cent. In small populations with a very low ARR frequency the ARR allele could be lost by genetic drift. A simulation study was therefore made to examine the effects of different breeding schemes in populations of different sizes on attempts to select for the ARR allele in an endangered population. In breeds in which no homozygous rams are available the breeding strategy would depend on the number of heterozygous rams, and the genotyping and selection of suitable breeding ewes would reduce the time required to achieve a highly resistant population. In general, in all the breeds a selection programme to achieve 99 per cent ARR homozygous genotypes would be feasible in six to nine generations, depending on the initial allele frequencies. In small populations the inbreeding rate may increase if no specific mating plans are developed by the breeding organisations.