Does covering of farm-associated Culicoides larval habitat reduce adult populations in the United Kingdom?

Culicoides biting midges (Diptera: Ceratopogonidae) are the biological vectors of a range of internationally important arboviruses of livestock, including bluetongue virus (BTV) and the recently emerging Schmallenberg virus (SBV). Culicoides species in the subgenus Avaritia (in the UK: Culicoides obsoletus Meigen, Culicoides scoticus Downes & Kettle, Culicoides dewulfi Goetghebuer and Culicoides chiopterus Meigen) have been implicated in BTV transmission in northern Europe and to a varying degree utilise cattle dung as a larval development substrate. The collection of cattle dung into heaps on farms provides a localised source of Culicoides emergence in close proximity to livestock. This study assesses the impact of covering dung heaps prior to the onset of adult Culicoides activity with the aim of reducing recruitment to the local adult populations at four livestock farms in England. Light suction trap catches of adult Culicoides from these farms were compared with those from four untreated control farms from a wide geographic range across the UK. It was demonstrated that implementing control of emergence from dung heaps did not have a significant impact upon the local adult subgenus Avaritia abundance at the treated farm holdings and that the onset of Culicoides activity was similarly unaffected. Use of this method in isolation is unlikely to have an effect in reducing the risk of BTV and SBV transmission. The implications of these results for control of farm-associated Culicoides in Europe are discussed.

Larval development and emergence sites of farm-associated Culicoides in the United Kingdom.

Culicoides biting midges (Diptera: Ceratopogonidae) are the biological vectors of internationally important arboviruses of livestock including bluetongue virus (BTV). Information on the habitats used by Culicoides for larval development is valuable for establishing targeted vector control strategies and for improving local scale models of vector abundance. This study combines emergence trap collections of adult Culicoides identified using molecular markers and physiochemical measurements of habitats to investigate larval development sites of Palaearctic Culicoides in South East England. The known range of larval habitats for several Culicoides species is extended and the potential BTV vector species C. obsoletus and C. scoticus are confirmed to co-occur in many larval habitats. The presence of emerging C. obsoletus was favoured by increasing substrate moisture level [odds ratio (OR) 6.94 (2.30; 20.90)] and substrate pH [OR 4.80 (1.66; 13.90)] [bias-corrected Dxy : 0.68; area under the curve (AUC): 0.86] rather than any particular larval habitat type, as expected for a species with relatively wide larval habitat preference. Of the newly emerged sub-genus Avaritia individuals collected, 23% were observed to have a degree of abdominal pigmentation commonly inferred to indicate parity. If consistent across species and locations, this observation represents a potential source of error for age structure analyses of Culicoides populations.

Investigating incursions of bluetongue virus using a model of long-distance Culicoides biting midge dispersal.

Bluetongue virus (BTV) is an economically important pathogen of ruminants that is the aetiological agent of the haemorrhagic disease bluetongue. Bluetongue virus is biologically transmitted by Culicoides biting midges (Diptera: Ceratopogonidae), and long-range dispersal of infected vector species contributes substantially to the rapid spread of the virus. The range of semi-passive flights of infected Culicoides on prevailing winds has been inferred to reach several hundred kilometres in a single night over water bodies. In this study, an atmospheric dispersion model was parameterized to simulate Culicoides flight activity based on dedicated entomological data sets collected in the UK. Five outbreaks of BTV in Europe were used to evaluate the model for use as an early warning tool and for retrospective analyses of BTV incursions. In each case, the generated predictions were consistent with epidemiological observations confirming its reliability for use in disease outbreak management. Furthermore, the model aided policy makers to predict, contain and eradicate BTV outbreaks in the UK during 2007 and 2008.

Collection of Culicoides (Diptera: Ceratopogonidae) using CO2 and enantiomers of 1-octen-3-ol in the United Kingdom.

The host kairomones carbon dioxide (CO2) and 1-octen-3-ol elicit a host seeking response in a wide range of haematophagous Diptera. This study investigates the response of Culicoides biting midges (Diptera: Ceratopogonidae) to these cues using field-based experiments at two sites in the United Kingdom with very different species complements. Traps used for surveillance (miniature CDC model 512) and control (Mosquito Magnet Pro) were modified to release ratios of (R)- and (S)-1-octen-3-ol enantiomers in combination with CO2 and, in the case of the latter trap type, a thermal cue. Abundance and species diversity were then compared between these treatments and against collections made using a trap with a CO2 lure only, in a Latin square design. In both habitats, results demonstrated that semiochemical lures containing a high proportion of the (R)-enantiomer consistently attracted a greater abundance of host-seeking Culicoides females than any other treatment. Culicoides collected using an optimal stimulus of 500 ml/min CO2 combined with 4.1 mg/h (R)-1-octen-3-ol were then compared with those collected on sheep through the use of a drop trap. While preliminary in nature, this trial indicated Culicoides species complements are similar between collections made using the drop trap in comparison to the semiochemical-baited CDC trap, and that there are advantages in using (R)-1-octen-3-ol.

Impacts of climate, host and landscape factors on Culicoides species in Scotland.

Culicoides biting midges (Diptera: Ceratopogonidae) vector a wide variety of internationally important arboviral pathogens of livestock and represent a widespread biting nuisance. This study investigated the influence of landscape, host and remotely-sensed climate factors on local abundance of livestock-associated species in Scotland, within a hierarchical generalized linear model framework. The Culicoides obsoletus group and the Culicoides pulicaris group accounted for 56% and 41%, respectively, of adult females trapped. Culicoides impunctatus Goetghebuer and C. pulicaris s.s. Linnaeus were the most abundant and widespread species in the C. pulicaris group (accounting for 29% and 10%, respectively, of females trapped). Abundance models performed well for C. impunctatus, Culicoides deltus Edwards and Culicoides punctatus Meigen (adjusted R(2) : 0.59-0.70), but not for C. pulicaris s.s. (adjusted R(2) : 0.36) and the C. obsoletus group (adjusted R(2) : 0.08). Local-scale abundance patterns were best explained by models combining host, landscape and climate factors. The abundance of C. impunctatus was negatively associated with cattle density, but positively associated with pasture cover, consistent with this species' preference in the larval stage for lightly grazed, wet rush pasture. Predicted abundances of this species varied widely among farms even over short distances (less than a few km). Modelling approaches that may facilitate the more accurate prediction of local abundance patterns for a wider range of Culicoides species are discussed.

Epizootic heamorragic disease.

Epizootic haemorrhagic disease (EHD) is an infectious non-contagious viral disease transmitted by insects of the genus Culicoides which affects wild and domestic ruminants. The causative agent, the epizootic haemorrhagic disease virus (EHDV), belongs to the family Reoviridae, genus Orbivirus and shares many morphological and structural characteristics with the other members of the genus such as bluetongue, African horse sickness and equine encephalosis viruses. In recent years EHD outbreaks have been reported in countries bordering the European Union. They caused disease in cattle and severe repercussion on the livestock industry of the affected countries. In the light of recent European bluetongue epizootic these events pose an increasing threat to the European Union. This review includes the most recent information regarding the virus and the disease as well as tools for its diagnosis and control. It is our conviction that more attention should be drawn to both EHDV and the disease itself in order to fulfil all these gaps and not to be unprepared in case future possible incursions.

A potential role for ixodid (hard) tick vectors in the transmission of lumpy skin disease virus in cattle.

Lumpy skin disease (LSD) is an economically important cattle disease. The disease is endemic in many African countries, but outbreaks have also been reported in Madagascar and the Middle East. The aim of this study was to investigate the potential role of ixodid (hard) ticks in the transmission of the disease. Cattle were infected with a virulent, South African field isolate of lumpy skin disease virus (LSDV). Three common African tick species (genera Rhipicephalus, Amblyomma and Rhipicephalus (Boophilus)) in different life cycle stages were fed on the infected animals during the viraemic stage and on skin lesions. Post-feeding, the partially fed male ticks were transferred to the skin of non-infected 'recipient' animals, while females were allowed to lay eggs that were then tested using the polymerase chain reaction (PCR) method and virus isolation. Nymphs were allowed to develop for 2-3 weeks after which time they were tested. The non-infected 'recipient' cattle were closely monitored, both skin and blood samples were tested using PCR and virus isolation, and serum samples were tested by the serum neutralization test. This is the first report showing molecular evidence of potential transmission of LSDV by ixodid ticks. The study showed evidence of transstadial and transovarial transmission of LSDV by R. (B.) decoloratus ticks and mechanical or intrastadial transmission by R. appendiculatus and A. hebraeum ticks.

Invasive arthropods.

Many arthropod species have been transported around the globe and successfully invaded new regions. Invasive arthropods can have severe impacts on animal and human health, agriculture and forestry, and the biodiversity of natural habitats as well as those modified by humans. The economic and environmental effects of invasion can be both direct, through feeding and competition, and indirect, such as the transmission of pathogens. In this paper, the authors consider ten examples that illustrate the main mechanisms of introduction, the characteristics that enable species to rapidly expand their ranges and some of the consequences of their arrival.

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.

Treatment of horses with cypermethrin against the biting flies Culicoides nubeculosus, Aedes aegypti and Culex quinquefasciatus.

An in vitro assay was used to assess the efficacy of the proprietary pyrethroid insecticide cypermethrin applied to horses (Deosect spray, 5.0%, w/v Fort Dodge Animal Health) against the biting midge Culicoides nubeculosus (Meigen) (Diptera: Ceratopogonidae) and the mosquitoes Aedes aegypti Linneaus and Culex quinquefasciatus Say (Diptera: Culicidae). Hair was collected from the back, belly and legs of the horses immediately prior to treatment and 7, 14, 21, 28 and 35 days after treatment, and also from untreated controls. In laboratory assays groups of 10 adult female C. nubeculosus, Ae. aegypti or C. quinquefasciatus were exposed to 0.5g of hair for 3min. In all cases, little or no mortality was observed in insects kept in contact with the pre-treatment samples or the untreated controls. With post-treatment samples for C. nubeculosus, mortality was close to 80% 7 days after treatment and then declined gradually; mean mortality was still at around 50% for hair collected 35 days after treatment. In general, Ae. aegypti and C. quinquefasciatus appeared to be less susceptible to cypermethrin than C. nubeculosus and the attenuation of the toxic effect declined more quickly with time after treatment. There were differences in the toxicity of hair from different body regions, with hair from the back consistently inducing the highest mortality and hair from the legs the lowest; this effect was more pronounced for C. nubeculosus than Ae. aegypti or C. quinquefasciatus. The results demonstrate the potential for topical insecticide treatment to offer protection to horses against biting flies; but highlight the major differences that exist in susceptibility between different insect species.

Insect bite hypersensitivity in the horse: comparison of IgE-binding proteins in salivary gland extracts from Simulium vittatum and Culicoides nubeculosus.

Insect bite hypersensitivity (IBH) is an IgE-mediated allergic dermatitis of horses caused by bites of insects such as Culicoides or Simulium spp. The aim of the present study was to compare the IgE-binding pattern of sera of IBH-affected horses to Culicoides nubeculosus and Simulium vittatum salivary gland extracts (SGE). Individual IgE responses to proteins of S. vittatum and C. nubeculosus SGEs were evaluated in 15 IBH-affected and three healthy horses on immunoblots. Fourteen out of the 15 IBH-affected but none of the healthy horses showed individual IgE binding patterns to seven and six main protein bands in C. nubeculosus and S. vittatum SGE, respectively. These 14 sera showed IgE-binding to proteins from SGE of both C. nubeculosus and S. vittatum, but they reacted with fewer protein bands derived from S. vittatum than from C. nubeculosus SGE. Sera showing IgE-binding to a 32 kDa band from C. nubeculosus always bound to a 32 kDa band from S. vittatum. Similarly, all sera binding to a 70 kDa band from C. nubeculosus reacted with a corresponding band in S. vittatum SGE. The 70 kDa bands from S. vittatum and C. nubeculosus were identified by mass spectrometry as heat shock protein-70-cognate-3.

Identification and isolation of cDNA clones encoding the abundant secreted proteins in the saliva proteome of Culicoides nubeculosus.

Culicoides spp. are vectors of several infectious diseases of veterinary importance and a major cause of allergy in horses and other livestock. Their saliva contains a number of proteins which enable blood feeding, enhance disease transmission and act as allergens. We report the construction of a novel cDNA library from Culicoides nubeculosus linked to the analysis of abundant salivary gland proteins by mass spectrometry. Fifty-four novel proteins sequences are described including those of the enzymes maltase, hyaluronidase and two serine proteases demonstrated to be present in Culicoides salivary glands, as well as several members of the D7 family and protease inhibitors with putative anticoagulant activity. In addition, several families of abundant proteins with unknown function were identified including some of the major candidate allergens that cause insect bite hypersensitivity in horses.

Life-history parameters of Culicoides (Avaritia) imicola Kieffer in the laboratory at different rearing temperatures.

This laboratory study investigates the sub-adult developmental cycle of field collected Culicoides (Avaritia) imicola Kieffer (Diptera; Ceratopogonidae). The period required from blood-feeding field-collected females to the production of progeny adults occupied 34-56 days at 20 degrees C, 15-21 days at 25 degrees C and 11-16 days at 28 degrees C, demonstrating clear temperature dependence. When reared at 28 degrees C, C. imicola demonstrated higher variability in fecundity (between 2.4 and 20.6 eggs/female) and lower hatching rates (50.0-62.2%), although larval survival rates to pupation were low at all temperatures (20-30%). Similarly, the mean emergence rate from pupae was the highest at lower temperatures. These results highlight the difficulty in establishing and maintaining a laboratory colony of this species from field-collected material and results are discussed in reference to future research directions that may aid this process.

Efficacy of alphacypermethrin applied to cattle and sheep against the biting midge Culicoides nubeculosus.

The recent emergence of bluetongue virus (BTV) in northern Europe, has led to an urgent need to identify methods to control the Culicoides biting midges that transmit it. Here, an in vitro assay was used to assess the effects of the proprietary pyrethroid insecticide alphacypermethrin applied to cattle and sheep (Dysect Cattle Pour-On, Dysect Sheep Pour-On; Ford Dodge Animal Health) against the biting midge Culicoides nubeculosus (Meigen) (Diptera: Ceratopogonidae). Hair or wool was collected from the back, belly and legs of animals immediately prior to treatment and 7, 14, 21, 28 and 35 days after treatment, and also from untreated controls. In the laboratory assay groups of 10 adult females C. nubeculosus were exposed to 0.5 g of hair or wool for 3 min. In all cases, no mortality was observed in the pre-treatment sample or the untreated controls. In the post-treatment samples, for both cattle and sheep mortality was close to 100% 7 days after treatment. For cattle, treatment effect persisted for up to 21 days post-treatment, following which the mortality rate following exposure to hair samples declined. In contrast, for sheep, mortality levels declined more slowly, and approximately 50% mortality was still observed 35 days after treatment. There was no significant difference in the kill rate for wool collected from the back, belly or legs of either sheep or cattle. The results demonstrate the potential for pour-on insecticide treatment to offer a degree of mitigation to livestock against onward transmission from infected animals--and in particular demonstrate that sufficient compound is able to reach the lower legs to kill in contact midges. The practical issues associated with achieving adequate protection are discussed.

Why were there no outbreaks of bluetongue in the UK during 2008?

With bluetongue rampant on the main European Continent in 2008, why were there no outbreaks reported in the UK? The essential criteria for introduction of the disease by carriage of infected midges on the wind have been assessed for 2006, 2007 and 2008, and it is concluded that temperatures were favourable for virus replication and midge activity and that suitable winds were present on a considerable number of occasions. A major difference between 2007, when virus was introduced to the UK, and 2008 was the extensive vaccination programme implemented by the UK Government in 2008, with the support of the farming community. Vaccination reduced the numbers of susceptible animals, making it difficult for a focus of disease to become established. The authors believe that if bluetongue re-establishes itself on the near Continent in 2009, it will be of critical importance that UK livestock are fully protected by vaccination against the disease.

Experimental infection studies of UK Culicoides species midges with bluetongue virus serotypes 8 and 9.

This paper describes a rapid, standardised method for testing the susceptibility to bluetongue virus (BTV) of northern Palaearctic Culicoides species midges that can be used to assess the competence of both field-caught and laboratory-infected midges. The method has been used to show that Culicoides scoticus can replicate btv serotype 8 and BTV serotype 9 strains to more than 3 log(10) TCID50/midge, the first evidence of the potential of this species to transmit BTV.

Control techniques for Culicoides biting midges and their application in the U.K. and northwestern Palaearctic.

The recent emergence of bluetongue virus (Reoviridae: Orbivirus) (BTV) in northern Europe, for the first time in recorded history, has led to an urgent need for methods to control the disease caused by this virus and the midges that spread it. This paper reviews various methods of vector control that have been employed elsewhere and assesses their likely efficacy for controlling vectors of BTV in northern Europe. Methods of controlling Culicoides spp. (Diptera: Ceratopogonidae) have included: (a) application of insecticides and pathogens to habitats where larvae develop; (b) environmental interventions to remove larval breeding sites; (c) controlling adult midges by treating either resting sites, such as animal housing, or host animals with insecticides; (d) housing livestock in screened buildings, and (e) using repellents or host kairomones to lure and kill adult midges. The major vectors of BTV in northern Europe are species from the Culicoides obsoletus (Meigen) and Culicoides pulicaris (L.) groups, for which there are scant data on breeding habits, resting behaviour and host-oriented responses. Consequently, there is little information on which to base a rational strategy for controlling midges or for predicting the likely impact of interventions. However, data extrapolated from the results of vector control operations conducted elsewhere, combined with some assessment of how acceptable or not different methods may be within northern Europe, indicate that the treatment of livestock and animal housing with pyrethroids, the use of midge-proofed stabling for viraemic or high-value animals and the promotion of good farm practice to at least partially eliminate local breeding sites are the best options currently available. Research to assess and improve the efficacy of these methods is required and, in the longer term, efforts should be made to develop better bait systems for monitoring and, possibly, controlling midges. All these studies will need better methods of analysing the ecology and behaviour of midges in the field than are currently in use. The paucity of control options and basic knowledge serve to warn us that we must be better prepared for the possible emergence of other midge-borne diseases, particularly African horse sickness.

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.