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In studies on Culicoides attacking livestock in the Netherlands, we chanced upon a species of the Obsoletus complex that we do not recognize, but whose dark wing pattern is distinctive. Nine cytochrome c oxidase (CO1) sequences of our so-called 'dark obsoletus' support its status as a separate species, the sequences differing significantly from those representing Culicoides obsoletus (Meigen) (90-91% homology) and Culicoides scoticus Downes & Kettle (87-88% homology). In the last decade, several research groups in Europe have encountered 'mystery species' related to C. obsoletus and in some instances have made their sequences for various genetic loci available in GenBank. These include a CO1 series submitted from Sweden in 2012 (annotated as 'obsoletus 01, 02, or 03 MA-2012') and of which some share a 99% identity with our sequences for 'dark obsoletus'. Without doubt, the series from the Netherlands, along with a portion of the Swedish submissions, together represent a single species ('dark obsoletus'). Whether this species is referable to the Russian Culicoides gornostaevae Mirzaeva recorded recently from Norway, Sweden and Poland, and based solely upon the external morphology of the male, is not clear. The presence in Western Europe of multiple undescribed species related to C. obsoletus means that the taxonomy of this important vector complex is not fully resolved; consequently, we know little about these cryptic species with regard to seasonality, geographic range and host preference. This is undesirable given that Culicoides-borne arboviruses causing disease in livestock are moving more regularly out of the tropics and spreading north into temperate latitudes., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Host preference is an important determinant of feeding behaviour in biting insects and a critical component in the transmission of vector-borne diseases. The aim of the study was to quantify Culicoides (Diptera: Ceratopogonidae) host preferences and biting rates using tethered livestock at pasture (a dairy cow and a sheep) and to compare the numbers of biting midges aspirated off them to those captured simultaneously in a black-light suction trap acting as a surrogate host. Culicoides collections were made hourly over seven hours (from five hours before official sunset to two hours after) between 27 May and 19 June, 2013 at a dairy farm (eastern Netherlands). The study involved 13 replicates of a site × host randomised design. Culicoides collected by black-light suction trap and by direct aspiration were identified to species morphologically and age-graded. The C. obsoletus complex, C. dewulfi and C. pulicaris predominated on the back and flanks of the animals, C. punctatus on the belly, and C. chiopterus on the legs. Using comparable collection periods, 9.3 times (95% confidence interval: 8.6-10.0) more Culicoides were caught on the cow than on the sheep and 25.4 times (95% confidence interval: 18.4-35.1) less in the black-light suction trap compared to the sheep. Mean Culicoides biting rates on the cow across the 7-h collection period were 4.6, 3.5, 1.0, 1.0 and 0.5 min(-1) for C. dewulfi, the C. obsoletus complex, C. chiopterus, C. punctatus and C. pulicaris, respectively; for the sheep they were 0.6, 0.4 and 0.1 min(-1) for the C. obsoletus complex, C. dewulfi and C. punctatus, respectively. Though midges were aspirated off livestock during each of the seven hours, they only began to appear in the black-light suction trap 5h later, from sunset onwards. After sunset, its efficacy improved markedly, but occurred when midge activity overall had begun to decline. Though it was quite accurate in ranking Culicoides species abundance, the black-light suction trap proved to be of limited value for determining hours of peak biting activity, levels of abundance, and host preference, in Culicoides., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Geographical maps indicating the value of the basic reproduction number, R₀, can be used to identify areas of higher risk for an outbreak after an introduction. We develop a methodology to create R₀ maps for vector-borne diseases, using bluetongue virus as a case study. This method provides a tool for gauging the extent of environmental effects on disease emergence. The method involves integrating vector-abundance data with statistical approaches to predict abundance from satellite imagery and with the biologically mechanistic modelling that underlies R₀. We illustrate the method with three applications for bluetongue virus in the Netherlands: 1) a simple R₀ map for the situation in September 2006, 2) species-specific R₀ maps based on satellite-data derived predictions, and 3) monthly R₀ maps throughout the year. These applications ought to be considered as a proof-of-principle and illustrations of the methods described, rather than as ready-to-use risk maps. Altogether, this is a first step towards an integrative method to predict risk of establishment of diseases based on mathematical modelling combined with a geographic information system that may comprise climatic variables, landscape features, land use, and other relevant factors determining the risk of establishment for bluetongue as well as of other emerging vector-borne diseases.

In 1999, among 164 randomly selected Dutch sheep farmers, a questionnaire was carried out to estimate the prevalence of myiasis in sheep and to investigate factors associated with the occurrence of myiasis. The total number of sheep and/or lambs on the reference date 1 August 1999 was 12,200: 5243 ewes, 225 rams, 3393 ewe- and 3339 ram lambs. On 86 (52.4%) of the farms 349 (2.9%) of all sheep and/or lambs contracted myiasis, of which two died. Myiasis was seen significantly more frequently on farms with over 25 ewes compared to smaller farms. Cases of myiasis were detected from April to September, with a peak (47.1%) in August. Occurrence of myiasis was most frequently associated with hot and humid weather and was mainly observed (69.1%) in the area around the tail.Ewes and ewe lambs had significantly more myiasis when compared with rams and ram lambs. There was no relationship with tail docking, with breed, with the time of shearing, with the kind of soil (clay, sand, etc.), with the environment (bush, trees, water, etc.), with the type of treatment (pour on, dipping, spraying), the used insecticides (synthetic pyrethroïds, diazinon, cyromazin, etc.), the number of preventive treatments, the time of treatment or the number of observations on the herd (once a day, once a week, etc.).

In this study, the rate of S. scabiei var. suis transmission among finishing pigs was quantified in a contact transmission experiment. Forty piglets originating from a mange free farrow-to-finish herd were randomly allocated to three groups and one S. scabiei var. suis infested finishing pig was subsequently added to each of these groups. After 35 days, the three seeder pigs were removed from the groups and the remaining 40 pigs were re-allocated to five pens. Ear scrapings, to be examined for mites, were collected from each pig on days 1, 14, 28, 42, 56, and 84 of the experiment. Blood samples, to be tested for antibodies against S. scabiei, were collected from each pig on days 0, 14, 28, 42, 56, 70, 84 and 112 after the introduction of the seeder pigs. From the results of the ear scrapings and the blood samples the number of susceptible (not infested) and infested pigs was derived at the time of each sample collection and the number of new infestations in the intervals between the sample collections. From these data the infestation rate parameter beta (average number of new infestations per infested pig per day) was estimated by use of a Generalised Linear Model (GLM) and accordingly, beta was estimated at 0.056 (95% CI: 0.037-0.085) infestations per infested pig per day.Next, by use of beta, the transmission of S. scabiei was simulated in a population of 100 finishing pigs for 100 days after the introduction of a single infested pig. For this purpose, 500 simulations were done. The 90% confidence interval of the number of infested pigs at day 100 ranged from 12 to 88 (median: 63). It was concluded that transmission of S. scabiei among finishing pigs is slow. Due to the presumed lower contact rate between sows as compared to finishing pigs, it is anticipated that transmission of S. scabiei among sows will even be slower than among finishers These findings are of particular interest for the development of surveillance programmes for S. scabiei free herds.

An Enzyme-linked ImmunoSorbent Assay (ELISA) was developed for the detection of antibodies to Sarcoptes scabiei. This 'Animal Health Service'-ELISA (AHS-ELISA) was compared with a commercial test (Checkit(R) Sarcoptest) using experimental and field sera. The experimental study was a contact infestation experiment. Eighty piglets were randomly divided between the experimental and control group. After introduction of three Sarcoptes scabiei var. suis infested pigs in the experimental group, both groups were monitored by determining scratching indices, taking ear scrapings and blood samples in Weeks 0, 2, 4, 6, 8, 12 and 16. Four pigs in the control group were immunised with either Dermatophagoides pteronyssinus (Dp) antigens (n=2), or Acarus siro (As) antigens (n=2). In the control group all (non-immunised) pigs were negative in all tests. In the experimental group only slightly elevated scratching indices were observed, with a maximum in Week 8. After 2 weeks for the first time an ear scraping was positive (2.5%). In Week 8 the highest number of positive ear scrapings were found (25.0%). Positive results in the Sarcoptest were first obtained in Week 12 (10.5% positive), while eventually 29.0% of the finishing pigs were positive after 16 weeks. The AHS-ELISA first detected a serological response after 6 weeks (5. 0% positives), increasing until after 16 weeks a large proportion (74.2%) of the finishing pigs were seropositive, making the AHS-ELISA the most sensitive test. In the AHS-ELISA one As-immunised pig remained seronegative, but the other hyper-immunised pigs crossreacted. In the Sarcoptest, only Dp-immunised pigs had elevated Optical Densities (OD's) albeit below the cut-off level. Although hyper-immunisation is not a representation of field conditions, it cannot be excluded that the AHS-ELISA is not 100% specific.Field samples were taken from 20 sows in 30 herds, classified as mange-free, suspect, or infested. On a herd level there was high agreement among the ELISAs. Both serological tests were suitable to distinguish mange-free herds from infested herds. In one infested herd the decline of maternal antibody in piglets was studied by sampling 40 piglets from 20 different litters. The lowest average OD using the AHS-ELISA was found at 5 weeks of age, followed by a significant increase at 7 weeks. The average OD with the Sarcoptest was at a minimum level at 3 weeks, but no increase was found later. For screening of herds, interference of maternal antibodies is avoided by sampling at an age of 7 weeks or older.

Haematological and clinico-chemical profiles of blood of healthy stress-resistant swine collected at the farm and at slaughter were determined to investigate whether values of blood variables can be used to establish stress. The values of most variables investigated showed highly significant changes. It is concluded that haematological and clinico-chemical values may be useful in studies to detect, quantify and reduce stress-provoking conditions in stress-resistant swine., (Copyright © 1992. Published by Elsevier Ltd.)