Identifying potential candidate Culicoides spp. for the study of interactions with Candidatus Cardinium hertigii.

Culicoides biting midges (Diptera: Ceratopogonidae) are vectors responsible for the transmission of several viruses of veterinary importance. Previous screens of Culicoides have described the presence of the endosymbiont Candidatus Cardinium hertigii (Bacteroidetes). However, any impacts of this microbe on vectorial capacity, akin to those conferred by Wolbachia in mosquitoes, are yet to be uncovered and await a suitable system to study Cardinium-midge interactions. To identify potential candidate species to investigate these interactions, accurate knowledge of the distribution of the endosymbiont within Culicoides populations is needed. We used conventional and nested PCR assays to screen Cardinium infection in 337 individuals of 25 Culicoides species from both Palearctic and Afrotropical regions. Infections were observed in several vector species including C. imicola and the Pulicaris complex (C. pulicaris, C. bysta, C. newsteadi and C. punctatus) with varying prevalence. Phylogenetic analysis based on the Gyrase B gene grouped all new isolates within 'group C' of the genus, a clade that has to date been exclusively described in Culicoides. Through a comparison of our results with previous screens, we suggest C. imicola and C. sonorensis represent good candidates for onward study of Cardinium-midge interactions.

The role of climate change in a developing threat: the case of bluetongue in Europe.

There is a solid theoretical basis for expecting climate change to have a considerable effect on the infectious diseases of humans, animals and plants. Vector-borne diseases are the most likely to be affected. It is, however, rare to observe such impacts, as diseases are also influenced by many other drivers, some of which may have stronger effects over shorter time scales than climate change. Nevertheless, there is evidence that our warming climate has already influenced some animal diseases, of which bluetongue is considered a prime example. Bluetongue emerged dramatically in southern Europe after 1998 and in northern Europe from 2006. While the speed and scale of this emergence is a challenge to explain, there is evidence, principally from the development of climate-driven models, that recent climate change has played a significant role. Climate-driven models point to an increase in the risk of bluetongue transmission in Europe in recent decades, caused by an increased suitability of parts of southern Europe for the Afro-tropical biting midge, Culicoides imicola, as well as an increase in the vectorial capacity of indigenous Culicoides vectors in northern Europe. Farm-to-farm transmission models of bluetongue in England and Wales under predicted climatic conditions further suggest that, under high-emission scenarios, the scale of future outbreaks could far exceed those experienced to date. The role of climate change in the developing threat of animal disease is, therefore, likely to be economically and socially costly, unless lower emission targets can be set and followed.

Nombre d'arguments théoriques sérieux confirment l'ampleur des effets du changement climatique sur les maladies infectieuses affectant l'être humain, les animaux et les végétaux. Les maladies à transmission vectorielle sont probablement les plus sujettes à cette influence. Toutefois, il est très rare de pouvoir observer ces effets directement, dans la mesure où d'autres facteurs exercent également une influence sur les maladies, dont certains ont des effets plus marquants et plus rapides que le changement climatique. L'influence du réchauffement climatique sur certaines maladies animales a néanmoins été prouvée ; à ce titre, le cas de la fièvre catarrhale ovine est considéré comme exemplaire. La fièvre catarrhale ovine a fait son apparition en Europe méridionale après 1998, puis en Europe du Nord à partir de 2006. Si la rapidité et l'envergure de cette émergence sont difficiles à expliquer, plusieurs démonstrations, recourant pour la plupart à des modèles axés sur le climat font état du rôle important joué par le changement climatique. Les modèles axés sur le climat font ressortir un risque accru de transmission de la fièvre catarrhale ovine en Europe au cours des dernières décennies, associé, d'une part, à l'adéquation croissante de régions entières de l'Europe méridionale vis-à-vis du moucheron afro-tropical Culicoides imicola et, d'autre part, à l'accroissement de la capacité vectorielle des vecteurs Culicoides autochtones dans le nord de l'Europe. D'après les études basées sur des modèles de transmission de la fièvre catarrhale ovine entre exploitations en Angleterre et au Pays de Galles, dans les conditions climatiques prévisibles, il apparaît qu'en cas de fortes émissions, l'ordre de grandeur des foyers futurs serait considérablement plus élevé que dans les épisodes que nous avons connus jusqu'à présent. Par conséquent, le rôle du changement climatique dans les menaces évolutives de santé animale risque d'avoir un coût économique et social élevé, à moins que des objectifs de réduction de l'émission soient mis en place et fassent l'objet d'un suivi approprié.

Existen sólidas bases teóricas para prever que el cambio climático tendrá efectos considerables en las enfermedades infecciosas que afectan al hombre, los animales o las plantas. Las que más probablemente se verán afectadas son las enfermedades transmitidas por vectores. Sin embargo, rara vez pueden observarse tales efectos, pues hay otros muchos factores que influyen en las enfermedades, algunos de los cuales, a una escala temporal más breve, pueden tener una influencia más marcada que el cambio climático. Aun así, hay pruebas de que el clima, en pleno proceso de calentamiento, ya ha incidido en algunas enfermedades animales, de las que la lengua azul se considera un perfecto ejemplo. La lengua azul hizo una espectacular aparición en el sur de Europa a partir de 1998, y en la Europa septentrional a partir de 2006. Aunque resulta difícil explicar la velocidad y las proporciones de tal aparición, existen sólidos indicios, obtenidos principalmente de la elaboración de modelos regidos por variantes climáticas, de que el reciente cambio climático ha cumplido una función importante. Estos modelos apuntan a un incremento del riesgo de transmisión de la lengua azul en Europa en los últimos decenios, lo que se explica por las condiciones más propicias al jején afrotropical, Culicoides imicola, que ofrecen ciertas partes de Europa meridional y por un aumento de la capacidad vectorial de los Culicoides autóctonos del norte de Europa. Los modelos de transmisión de la lengua azul entre explotaciones agropecuarias de Inglaterra y Gales en las condiciones climáticas predichas indican además que, en la hipótesis de un elevado volumen de emisiones, los futuros brotes pueden revestir una escala muy superior a cuanto hemos conocido hasta ahora. Por consiguiente, a menos que se logre establecer y cumplir objetivos de emisiones menos cuantiosas, es probable que el cambio climático resulte económica y socialmente gravoso por su incidencia en la creciente amenaza que plantean las enfermedades animales.

Evaluation of a temperate climate mosquito, Ochlerotatus detritus (=Aedes detritus), as a potential vector of Japanese encephalitis virus.

The U.K. has not yet experienced a confirmed outbreak of mosquito-borne virus transmission to people or livestock despite numerous autochthonous epizootic and human outbreaks of mosquito-borne diseases on the European mainland. Indeed, whether or not British mosquitoes are competent to transmit arboviruses has not been established. Therefore, the competence of a local (temperate) British mosquito species, Ochlerotatus detritus (=Aedes detritus) (Diptera: Culicidae) for transmission of a member of the genus Flavivirus, Japanese encephalitis virus (JEV) as a model for mosquito-borne virus transmission was assessed. The JEV competence in a laboratory strain of Culex quinquefasciatus (Diptera: Culicidae), a previously incriminated JEV vector, was also evaluated as a positive control. Ochlerotatus detritus adults were reared from field-collected juvenile stages. In oral infection bioassays, adult females developed disseminated infections and were able to transmit virus as determined by the isolation of virus in saliva secretions. When pooled at 7-21 days post-infection, 13% and 25% of O. detritus were able to transmit JEV when held at 23 °C and 28 °C, respectively. Similar results were obtained for C. quinquefasciatus. To our knowledge, this study is the first to demonstrate that a British mosquito species, O. detritus, is a potential vector of an exotic flavivirus.

First detection of endosymbiotic bacteria in biting midges Culicoides pulicaris and Culicoides punctatus, important Palaearctic vectors of bluetongue virus.

Heritable bacteria have been highlighted as important components of vector biology, acting as required symbionts with an anabolic role, altering competence for disease transmission, and affecting patterns of gene flow by altering cross compatibility. In this paper, we tested eight U.K. species of Culicoides (Diptera: Ceratopogonidae) midge for the presence of five genera of endosymbiotic bacteria: Cardinium (Bacteroidales: Bacteroidaceae); Wolbachia (Rickettsiales: Rickettsiaceae); Spiroplasma (Entomoplasmatales: Spiroplasmataceae); Arsenophonus (Enterobacteriales: Enterobacteriaceae), and Rickettsia (Rickettsiales: Rickettsiaceae). Cardinium spp. were detected in both sexes of Culicoides pulicaris and Culicoides punctatus, two known vectors of bluetongue virus. Cardinium spp. were not detected in any other species, including the Culicoides obsoletus group, the main vector of bluetongue and Schmallenberg viruses in northern Europe. The other endosymbionts were not detected in any Culicoides species. The Cardinium strain detected in U.K. Culicoides species is very closely related to the Candidatus Cardinium hertigii group C, previously identified in Culicoides species in Asia. Further, we infer that the symbiont is not a sex ratio distorter and shows geographic variation in prevalence within a species. Despite its detection in several species of Culicoides that vector arboviruses worldwide, the absence of Cardinium in the C. obsoletus group suggests that infections of these symbionts may not be necessary to the arboviral vector competence of biting midges.

The influence of host number on the attraction of biting midges, Culicoides spp., to light traps.

A preliminary study was undertaken to investigate how the number of sheep below a light-suction trap affects the number of female Culicoides obsoletus Meigen (Diptera: Ceratopogonidae) caught. As the number of sheep increased from zero to three, the number of midges caught increased, but there appeared to be no further increase when six sheep were used. The lack of increase between three and six sheep is attributable to different activity rates on certain nights, perhaps in response to weather, and suggests, therefore, that catches in light traps increase linearly with sheep numbers, at least for small host numbers.

The impact of sheep breed on the risk of classical scrapie.

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

Evaluation of housing as a means to protect cattle from Culicoides biting midges, the vectors of bluetongue virus.

The housing of animals at night was investigated as a possible means of protecting them from attack by Culicoides biting midges (Diptera: Ceratopogonidae), the vectors of bluetongue. Light-trap catches of Culicoides were compared inside and outside animal housing, in the presence and absence of cattle. A three-replicate, 4 x 4 Latin square design was used at four farms in Bala, north Wales, over 12 nights in May and June 2007, and the experiment repeated in October. In the two studies, respectively, >70 000 and >4500 Culicoides were trapped, of which 93% and 86%, respectively, were of the Culicoides obsoletus group. Across the four farms, in May and June, the presence of cattle increased catches of C. obsoletus by 2.3 times, and outside traps caught 6.5 times more insects than inside traps. Similar patterns were apparent in October, but the difference between inside and outside catches was reduced. Catches were strongly correlated with minimum temperature and maximum wind speed and these two variables explained a large amount of night-to-night variation in catch. Outside catches were reduced, to a greater extent than inside catches, by colder minimum temperatures and higher maximum wind speeds. These conditions occur more frequently in October than in May and June, thereby suppressing outside catches more than inside catches, and reducing the apparent degree of exophily of C. obsoletus in autumn. The results suggest that the risk of animals receiving bites from C. obsoletus is reduced by housing at both times of year and the benefit would be greatest on warm, still nights when outside catches are at their greatest.

The UK's suitability for Aedes albopictus in current and future climates.

The Asian tiger mosquito Aedes albopictus is able to transmit various pathogens to humans and animals and it has already caused minor outbreaks of dengue and chikungunya in southern Europe. Alarmingly, it is spreading northwards and its eggs have been found in the UK in 2016 and 2017. Climate-driven models can help to analyse whether this originally subtropical species could become established in northern Europe. But so far, these models have not considered the impact of the diurnal temperature range (DTR) experienced by mosquitoes in the field. Here, we describe a dynamical model for the life cycle of Ae. albopictus, taking into account the DTR, rainfall, photoperiod and human population density. We develop a new metric for habitat suitability and drive our model with different climate data sets to analyse the UK's suitability for this species. For now, most of the UK seems to be rather unsuitable, except for some densely populated and high importation risk areas in southeast England. But this picture changes in the next 50 years: future scenarios suggest that Ae. albopictus could become established over almost all of England and Wales, indicating the need for continued mosquito surveillance.

The effect of temperature, farm density and foot-and-mouth disease restrictions on the 2007 UK bluetongue outbreak.

In 2006, bluetongue (BT), a disease of ruminants, was introduced into northern Europe for the first time and more than two thousand farms across five countries were affected. In 2007, BT affected more than 35,000 farms in France and Germany alone. By contrast, the UK outbreak beginning in 2007 was relatively small, with only 135 farms in southeast England affected. We use a model to investigate the effects of three factors on the scale of BT outbreaks in the UK: (1) place of introduction; (2) temperature; and (3) animal movement restrictions. Our results suggest that the UK outbreak could have been much larger had the infection been introduced into the west of England either directly or as a result of the movement of infected animals from southeast England before the first case was detected. The fact that air temperatures in the UK in 2007 were marginally lower than average probably contributed to the UK outbreak being relatively small. Finally, our results indicate that BT movement restrictions are effective at controlling the spread of infection. However, foot-and-mouth disease restrictions in place before the detection and control of BT in 2007 almost certainly helped to limit BT spread prior to its detection.

Rates of bluetongue virus transmission between Culicoides sonorensis and sheep.

Two experiments were undertaken to estimate the transmission rates of bluetongue virus (BTV) serotype 1 between a biting midge vector, Culicoides sonorensis (Wirth & Jones) (Ceratopogonidae), and a natural host, sheep. In an experiment to measure the transmission rate from vector to host (V-->H), six batches of one, five and 20 intrathoracically infected midges were fed on a total of 18 bluetongue (BT)-naïve sheep. The sheep were then monitored for 21 days for clinical signs of BT, viraemia and antibody response. All sheep fed on by five or 20 midges and five of six sheep fed on by just one midge showed signs of BT, were viraemic and developed antibody. The sixth sheep fed on by a single infected midge did not show signs of BT or have detectable viraemia; it did, however, develop a weak antibody response. A bite from a single infected midge is therefore able to transmit BTV to naïve sheep with 80-100% efficiency. Sheep fed upon by larger numbers of infected midges took less time to reach maximum viraemia and developed stronger antibody responses. Sheep exposed to greater amounts of BTV in feeding midges developed a higher level of viraemia and stronger antibody responses. In a second experiment to measure the transmission rate from host to vector (H-->V), batches of up to 500 uninfected female C. sonorensis fed every 1-2 days on two experimentally infected sheep during the course of infection. Of 3929 engorged midges that were individually titrated after surviving the extrinsic incubation period, only 23 (0.6%) were infected with BTV. Viraemia in the sheep extended for up to 19 days post-inoculation. No infected midges, however, were detected from 14 days post-infection.

The challenges posed by equine arboviruses.

Equine populations worldwide are at increasing risk of infection by viruses transmitted by biting arthropods, including mosquitoes, biting midges (Culicoides), sandflies and ticks. These include the flaviviruses (Japanese encephalitis, West Nile and Murray Valley encephalitis), alphaviruses (eastern, western and Venezuelan encephalitis) and the orbiviruses (African horse sickness and equine encephalosis). This review provides an overview of the challenges faced in the surveillance, prevention and control of the major equine arboviruses, particularly in the context of these viruses emerging in new regions of the world.

Potential vectors of equine arboviruses in the UK.

There is growing concern about the increasing risk of disease outbreaks caused by arthropod-borne viruses (arboviruses) in both human beings and animals. There are several mosquito-borne viral diseases that cause varying levels of morbidity and mortality in horses and that can have substantial welfare and economic ramifications. While none has been recorded in the UK, vector species for some of these viruses are present, suggesting that UK equines may be at risk. The authors undertook, therefore, the first study of mosquito species on equine premises in the UK. Mosquito magnet traps and red-box traps were used to sample adults, and larvae were collected from water sources such as tyres, buckets, ditches and pools. Several species that are known to be capable of transmitting important equine infectious arboviruses were trapped. The most abundant, with a maximum catch of 173 in 72 hours, was Ochlerotatus detritus, a competent vector of some flaviviruses; the highest densities were found near saltmarsh habitats. The most widespread species, recorded at >75 per cent of sites, was Culiseta annulata. This study demonstrates that potential mosquito vectors of arboviruses, including those known to be capable of infecting horses, are present and may be abundant on equine premises in the UK.

Descriptive analysis of the results of an anonymous postal survey of the occurrence of scrapie in Great Britain in 2002.

An anonymous postal survey was conducted in 2002 to estimate the proportion of farms in Great Britain affected with scrapie and to gather information on the likely risk factors for the occurrence of the disease; the response rate was 53 per cent. The survey showed that 1 per cent of the respondents thought they had had scrapie in their flock in the previous 12 months, and that 12 per cent thought they had had scrapie in the past. The results of the survey were consistent with the results of a similar survey carried out in 1998, and with notification patterns, but in 1998 approximately 3 per cent of farmers reported having had scrapie in the previous 12 months. It is not clear whether the apparent decrease in the prevalence of scrapie is real or whether it may be due to factors such as sampling biases, or to the increasing knowledge of the signs of scrapie shown by the respondents in 2002.

African horse sickness: The potential for an outbreak in disease-free regions and current disease control and elimination techniques.

African horse sickness (AHS) is an arboviral disease of equids transmitted by Culicoides biting midges. The virus is endemic in parts of sub-Saharan Africa and official AHS disease-free status can be obtained from the World Organization for Animal Health on fulfilment of a number of criteria. AHS is associated with case fatality rates of up to 95%, making an outbreak among naïve horses both a welfare and economic disaster. The worldwide distributions of similar vector-borne diseases (particularly bluetongue disease of ruminants) are changing rapidly, probably due to a combination of globalisation and climate change. There is extensive evidence that the requisite conditions for an AHS epizootic currently exist in disease-free countries. In particular, although the stringent regulations enforced upon competition horses make them extremely unlikely to redistribute the virus, there are great concerns over the effects of illegal equid movement. An outbreak of AHS in a disease free region would have catastrophic effects on equine welfare and industry, particularly for international events such as the Olympic Games. While many regions have contingency plans in place to manage an outbreak of AHS, further research is urgently required if the equine industry is to avoid or effectively contain an AHS epizootic in disease-free regions. This review describes the key aspects of AHS as a global issue and discusses the evidence supporting concerns that an epizootic may occur in AHS free countries, the planned government responses, and the roles and responsibilities of equine veterinarians.

Morphometric discrimination of two sympatric sibling species in the Palaearctic region, Culicoides obsoletus Meigen and C. scoticus Downes & Kettle (Diptera: Ceratopogonidae), vectors of bluetongue and Schmallenberg viruses.

BACKGROUND: Some Palaearctic biting midge species (subgenus Avaritia) have been implicated as vectors of bluetongue virus in northern Europe. Separation of two species (C. obsoletus and C. scoticus) is considered difficult morphologically and, often, these female specimens are grouped in entomological studies. However, species-specific identification is desirable to understand their life history characteristics, assess their roles in disease transmission or measure their abundance during arboviral outbreaks. This study aims to investigate whether morphometric identification techniques can be applied to female C. obsoletus and C. scoticus individuals trapped at different geographical regions and time periods during the vector season. METHODS: C. obsoletus and C. scoticus were collected using light-suction traps from the UK, France and Spain, with two geographical locations sampled per country. A total of 759 C. obsoletus/C. scoticus individuals were identified using a molecular assay based on the cytochrome c oxidase subunit I gene. Fifteen morphometric measurements were taken from the head, wings and abdomen of slide-mounted specimens, and ratios calculated between these measurements. Multivariate analyses explored whether a combination of morphometric variables could lead to accurate species identification. Finally, Culicoides spp. collected in France at the start, middle and end of the adult vector season were compared, to determine whether seasonal variation exists in any of the morphometric measurements. RESULTS: The principal component analyses revealed that abdominal characteristics: length and width of the smaller and larger spermathecae, and the length of the chitinous plates and width between them, are the most reliable morphometric characteristics to differentiate between the species. Seasonal variation in the size of each species was observed for head and wing measurements, but not abdominal measurements. Geographical variation in the size of Culicoides spp. was also observed and is likely to be related to temperature at the trapping sites, with smaller individuals trapped at more southern latitudes. CONCLUSIONS: Our results suggest that female C. obsoletus and C. scoticus individuals can be separated under a stereomicroscope using abdominal measurements. Although we show the length and width of the spermathecae can be used to differentiate between the species, this can be time-consuming, so we recommend undertaking this using standardized subsampling of catches.

Comparison of the h-Index Scores Among Pathogens Identified as Emerging Hazards in North America.

Disease surveillance must assess the relative importance of pathogen hazards. Here, we use the Hirsch index (h-index) as a novel method to identify and rank infectious pathogens that are likely to be a hazard to human health in the North American region. This bibliometric index was developed to quantify an individual's scientific research output and was recently used as a proxy measure for pathogen impact. Analysis of more than 3000 infectious organisms indicated that 651 were human pathogen species that had been recorded in the North American region. The h-index of these pathogens ranged from 0 to 584. The h-index of emerging pathogens was greater than non-emerging pathogens as was the h-index of frequently pathogenic pathogens when compared to non-pathogenic pathogens. As expected, the h-index of pathogens varied over time between 1960 and 2011. We discuss how the h-index can contribute to pathogen prioritization and as an indicator of pathogen emergence.

The genetics of scrapie in sheep and goats.

Scrapie, an invariably fatal disease of sheep and goats, is a transmissible spongiform encephalopathy (TSE). The putative infectious agent is the host-encoded prion protein, PrP. The development of scrapie is closely linked to polymorphisms in the host PrP gene. The pathogenesis of most TSEs involves conversion of normal, cellular PrP into a protease-resistant, pathogenic isoform called PrPSc. The conversion to PrPSc involves change in secondary structure; it is impacts on these structural changes that may link polymorphisms to disease. Within the structured C-terminal part of PrP polymorphisms have been reported at 15 and 10 codons of the sheep and goat PrP genes respectively. Three polymorphisms in sheep are acutely linked to the occurrence of scrapie: A136V, R154H and Q171R/H. These generate five commonly observed alleles: ARQ, ARR, AHQ, ARH and VRQ. ARR and AHQ are associated with resistance; ARQ, ARH and VRQ are associated with susceptibility. There are subtle effects of specific allele pairings (genotypes). Generally, more susceptible genotypes have younger ages at death from scrapie. Different strains of scrapie occur which may attack genotypes differently. Different sheep breeds vary in the assortment of the five alleles that they predominantly encode. The reason for this variation is not known. Furthermore, certain genotypes may be susceptible to scrapie in some breeds and resistant in others. The explanation is not known, but may relate to different scrapie strains circulating in different breeds, or there may be effects of other genes which modulate the effect of PrP.

Repellent effect of topical deltamethrin on blood feeding by Culicoides on horses.

African horse sickness (AHS) is a vectorborne disease spread by Culicoides biting midges. The UK's Department for Environment, Food and Rural Affairs currently suggests using topical deltamethrin for AHS control; however, no data are available regarding its efficacy in the horse. The aims of this study were to investigate the effect of topical deltamethrin on blood feeding by Culicoides on horses and to investigate which Culicoides species blood fed on horses. Three pairs of horses were placed in partially enclosed cages that allowed samples representing the Culicoides interacting with individual horses to be sampled. Four data collection sessions were completed before one horse from each pair was topically treated with 10 ml of 1 per cent deltamethrin solution and another four sessions were then carried out. Collected Culicoides were identified and each biting midge examined to see if it had blood fed. The most abundant species collected were C. chiopterus, C. dewulfi, C. obsoletus and C. scoticus (44.3 per cent) and either C. pulicaris or C. punctatus (34.7 per cent). These species were also more likely to have blood fed than other species, supporting their potential role as AHS vectors if the virus were to reach the UK. There was no significant effect of treatment on blood feeding by Culicoides. The results do not support the use of topical deltamethrin to prevent blood feeding by Culicoides on individual horses; however, the study does not investigate the effect that the widespread use of topical deltamethrin might have on vector numbers or disease transmission from viraemic individuals during an outbreak of AHS.

The signature of scrapie: differences in the PrP genotype profile of scrapie-affected and scrapie-free UK sheep flocks.

The amino-acid sequence of the PrP protein plays an important role in determining whether sheep are susceptible to scrapie. Although the genetics of scrapie susceptibility are now well understood, there have been few studies of the PrP gene at the population level, especially in commercially farmed sheep. Here we describe the PrP genetic profiles of the breeding stock of four UK sheep flocks, comprising nearly 650 animals in total. Two flocks had been scrapie affected for about eight years and two were scrapie free. Scrapie-resistant PrP genotypes predominated in all flocks but highly susceptible genotypes were present in each case. The distribution of PrP genotypes was similar in the scrapie-affected and scrapie-free flocks. The former, however, showed a slight but significant skew towards more susceptible genotypes despite their previous losses of susceptible sheep. Surprisingly, this skew was apparent in younger, but not older, sheep. We suggest that these patterns may occur if sheep flocks destined to become scrapie affected are predisposed by a genetic profile skewed towards susceptibility. The age structure of the scrapie-affected flocks suggests that the number of losses attributable directly or indirectly to scrapie considerably exceeds that recognized by the farmers, and also that significant losses may occur even in sheep of a moderately susceptible genotype. Similar patterns were not detected in the scrapie-free flocks, indicating that these losses are associated with scrapie infection as well as genotype.