Modelling African horse sickness emergence and transmission in the South African control area using a deterministic metapopulation approach.

African horse sickness is an equine orbivirus transmitted by Culicoides Latreille biting midges. In the last 80 years, it has caused several devastating outbreaks in the equine population in Europe, the Far and Middle East, North Africa, South-East Asia, and sub-Saharan Africa. The disease is endemic in South Africa; however, a unique control area has been set up in the Western Cape where increased surveillance and control measures have been put in place. A deterministic metapopulation model was developed to explore if an outbreak might occur, and how it might develop, if a latently infected horse was to be imported into the control area, by varying the geographical location and months of import. To do this, a previously published ordinary differential equation model was developed with a metapopulation approach and included a vaccinated horse population. Outbreak length, time to peak infection, number of infected horses at the peak, number of horses overall affected (recovered or dead), re-emergence, and Rv (the basic reproduction number in the presence of vaccination) were recorded and displayed using GIS mapping. The model predictions were compared to previous outbreak data to ensure validity. The warmer months (November to March) had longer outbreaks than the colder months (May to September), took more time to reach the peak, and had a greater total outbreak size with more horses infected at the peak. Rv appeared to be a poor predictor of outbreak dynamics for this simulation. A sensitivity analysis indicated that control measures such as vaccination and vector control are potentially effective to manage the spread of an outbreak, and shortening the vaccination window to July to September may reduce the risk of vaccine-associated outbreaks.

Revision of the Culicoides (Avaritia) Imicola complex Khamala & Kettle (Diptera: Ceratopogonidae) from the Australasian region.

The monophyly of the Imicola complex, a natural species complex within subgenus C. subgen. Avaritia Fox of the biting midge genus Culicoides Latreille, is supported using morphological and molecular analyses. A diagnosis for the group along with comparative redescriptions of the male and female of the species represented in Australasia, C. brevitarsis Kieffer and C. nudipalpis Delfinado and a description of C. asiatica Bellis sp. nov., are presented together with keys for their specific determination and molecular support for their status.

Description of Culicoides truuskae sp. n. (Diptera: Ceratopogonidae) from southern Africa.

Culicoides truuskae Labuschagne and Meiswinkel sp. n. is described and illustrated in both sexes from material collected in South Africa and Namibia. It is restricted to the xeric western margin of the subcontinent, occurring in Fynbos, Nama-Karoo and Succulent Karoo ecoregions in South Africa and Desert and Savanna ecoregions in Namibia experiencing 600 mm of rainfall annually. Culicoides truuskae sp. n. is part of the Afrotropical 'plain-wing' Culicoides in which the wing lacks a distinguishing pattern of light and dark spots; the diagnostic dark smudge that traverses wing cell r3 may result in C. truuskae sp. n. being misidentified as the sympatric but phyletically unrelated Culicoides herero (Enderlein) - (of the Similis group, subgenus Oecacta Poey). Additionally, this study is the first description of the male of C. herero. C. truuskae sp. n. and Culicoides coarctatus Clastrier and Wirth share similar characters in the male genitalia, although the two species are separable on wing pattern and female flagellum sensilla coeloconica (SCo) distribution. The breeding habitat and adult female blood-feeding preferences of C. truuskae sp. n. are not known. A maximum likelihood phylogenetic tree, using mitochondrial cytochrome c oxidase I (COI) sequence data, is provided to further clarify the relationship between C. truuskae sp. n., C. coarctatus and C. herero. Extensive light trap data, collected over 30 years, are used to map the distribution ranges of C. truuskae sp. n., C. coarctatus and C. herero in Southern Africa.Contribution: The description of this new species and the description of the male of C. herero increases our understanding of the diversity and distribution of Culicoides species in southern Africa.

The utilisation of CytB and COI barcodes for the identification of bloodmeals and Culicoides species (Diptera: Ceratopogonidae) reveals a variety of novel wildlife hosts in South Africa.

Biting midges in the genus Culicoides (Diptera; Ceratopogonidae) are vectors of pathogens that can cause diseases of major economic importance in humans and animals. Identifying host ranges of these biting midges might aid in understanding the complex epidemiology of such diseases, often involving reservoir hosts and multiple species. In this study, we aim to identify bloodmeal origin from engorged female Culicoides biting midges. All bloodfed females were opportunistically collected as part of an ongoing surveillance programme using Onderstepoort light traps in two provinces in South Africa. DNA of individuals was extracted and subjected to PCR targeting the cytochrome B (CytB) gene region of mammals and avians as well as cytochrome oxidase I (COI) for species identification. In total, 21 new reference barcodes were generated for C. bedfordi, C imicola, C. leucosticus, C. magnus, and C. pycnostictus. Seventy-four blood meals were identified, originating from 12 mammal and three avian species. COI sequence data performed well for species delimitation and 54 Culicoides specimens were identified with C. imicola the predominant species identified (41.8%). Generally, Culicoides species feed on a variety of hosts and host availability might be an important factor when selecting a host. Culicoides species thus appear to be opportunistic feeders rather than specialists. This implicates Culicoides as transfer vectors and demonstrates possible transmission routes of arboviruses and other pathogens from wildlife onwards to domestic animals and humans.

Molecular genotyping and epidemiology of equine piroplasmids in South Africa.

Recently reported substantial genetic diversity within Theileria equi 18S rRNA gene sequences has led to the identification of five genotypes A, B, C, D, and E, complicating molecular and serological diagnosis. In addition, T. haneyi has lately been reported as a species closely related to the T. equi 18S rRNA genotype C (Knowles et al., 2018). Theileria spp. of this group have a monophyletic origin and are therefore referred to as Equus group to distinguish them from the remaining Theileria lineages (Jalovecka et al., 2019). In this study, we report on the development of genotype-specific quantitative real-time PCR assays capable of detecting and distinguishing between each parasite genotype. Alignment of complete 18S rRNA sequences available on GenBank allowed for the design of a single primer pair and five TaqMan minor groove binder (MGB™) probes specific for each genotype (A-E). The assays, evaluated as qPCR simplex and two qPCR multiplex formats (Multiplex EP-ABC and Multiplex EP-DE), were shown to be both efficient and specific in the detection of T. equi genotypes. The developed qPCR assays were used to study (i) the intra-specific diversity of parasite genotypes within horse and zebra, (ii) the inter-specific differences in parasite genotype diversity in horses as compared to zebra, and (iii) the geographic distribution of T. equi 18S rRNA genotypes in South Africa. In addition, (iv) the presence of T. haneyi in South Africa was evaluated. An assessment of 342 equine field samples comprising 149 field horses, 55 racehorses, and 138 wild zebra confirmed the previously reported presence of T. equi 18S rRNA genotypes A, B, C, and D, and absence of genotype E in South African equids. Theileria equi genotypes A, B, C, and D, were detected in zebra, whereas only genotypes A, C and D, could be identified in field horses, and only genotypes A and C in racehorses. Genotypes B and D were the dominant genotypes identified in zebra in South Africa, while horses were predominantly infected with T. equi genotypes A and C. The greater diversity of T. equi genotypes in zebra suggests that it is an ancestral host for this piroplasmid lineage. Importantly, evidence is presented that each identified T. equi genotype segregates independently in each of the three studied equid populations reinforcing the notion that they represent individual separate entities corresponding to species. Preliminary investigations of the relationship between T. equi genotype C infections and Theileria haneyi, suggest that in addition to the five currently known T. equi genotypes, South African equids are also infected with T. haneyi.

Culicoides Latreille in the sun: faunistic inventory of Culicoides species (Diptera: Ceratopogonidae) in Mayotte (Comoros Archipelago, Indian Ocean).

BACKGROUND: The south-west insular territories of the Indian Ocean have recently received attention concerning the diversity of arthropods of medical or veterinary interest. While a recent study highlighted the circulation of Culicoides-borne viruses, namely bluetongue and epizootic hemorrhagic disease, with clinical cases in Mayotte (comprising two islands, Petite-Terre and Grand-Terre), Comoros Archipelago, no data have been published concerning the species diversity of Culicoides present on the two islands. RESULTS: A total of 194,734 biting midges were collected in 18 sites, covering two collection sessions (April and June) in Mayotte. Our study reports for the first time livestock-associated Culicoides species and recorded at least 17 described Afrotropical species and one undescribed species. The most abundant species during the April collection session were C. trifasciellus (84.1%), C. bolitinos (5.4%), C. enderleini (3.9%), C. leucostictus (3.3%) and C. rhizophorensis (2.1%). All other species including C. imicola represented less than 1% of the total collection. Abundance ranged between 126-78,842 females with a mean and median abundance of 14,338 and 5111 individuals/night/site, respectively. During the June collection, the abundance per night was low, ranging between 6-475 individuals. Despite low abundance, C. trifasciellus and C. bolitinos were still the most abundant species. Culicoides sp. #50 is recorded for the first time outside South Africa. CONCLUSIONS: Our study reports for the first time the Culicoides species list for Mayotte, Comoros Archipelago, Indian Ocean. The low abundance and rare occurrence of C. imicola, which is usually considered the most abundant species in the Afrotropical region, is unexpected. The most abundant and frequent species is C. trifasciellus, which is not considered as a vector species so far, but its role needs further investigation. Further work is needed to describe Culicoides sp. #50 and to carry on faunistic investigations on the other islands of the archipelago as well as in neighboring countries.

A field investigation of an African horse sickness outbreak in the controlled area of South Africa in 2016.

An outbreak of African horse sickness (AHS) caused by AHS virus type 1 occurred within the South African AHS surveillance zone during April and May 2016. The index case was detected by a private veterinarian through passive surveillance. There were 21 cases in total, which is relatively low compared to case totals during prior AHS outbreaks in the same region (and of the same AHS virus type) in 2004, 2011 and 2014. The affected proportion of horses on affected properties was 0.07 (95% CI 0.04, 0.11). Weather conditions were conducive to high midge activity immediately prior to the outbreak but midge numbers decreased rapidly with the advent of winter. The outbreak was localized, with 18 of the 21 cases occurring within 8 km of the index property and the three remaining cases on two properties within 21 km of the index property, with direction of spread consistent with wind-borne dispersion of infected midges. Control measures included implementation of a containment zone with movement restrictions on equids. The outbreak was attributed to a reversion to virulence of a live attenuated vaccine used extensively in South Africa. Outbreaks in the AHS control zones have a major detrimental impact on the direct export of horses from South Africa, notably to the European Union.

Risk of establishment of canine leishmaniasis infection through the import of dogs into South Africa.

Canine leishmaniasis is a vector-borne disease caused by protozoa of the genus Leishmania that affect dogs, humans and wildlife. Sandflies of the genera Phlebotomus and Lutzomyia are the primary vectors. Canine leishmaniasis is an exotic and controlled disease in South Africa. The main purpose of our risk assessment study was to evaluate the likelihood that this exotic disease could enter and be established in South Africa through importation of live dogs. Risk analysis to the spread of the disease follows the World Organization for Animal Health (OIE) formal method of quantitative risk assessment documented as a step-by-step process. We have identified and discussed 11 possible risk factors involved in three steps for final assessment. The annual average number of diagnostic tests performed on imported dogs from 44 countries for 2011-2015 was 1158. Leishmania is reported to occur in 21/44 (47.7%) exporting countries. A total of 71.1% of Leishmania positive dogs were imported from these endemic countries. The yearly percentage of Leishmania positive dogs ranged from 0.2% to 2%. Three confirmed clinical and fatal cases of leishmaniasis in dogs of unidentified origin have been reported by our laboratory and the state veterinarians. The disease has been reported in neighbouring countries as well as the putative sandfly vectors. This study concluded that the risk for the introduction and degree of uncertainty of Leishmania in imported dogs in South Africa are moderate. Risk mitigation and recommendations such as investigations into possible occurrence of autochthonous leishmaniasis in the country, surveillance in its wildlife reservoirs and systematic surveillance of sandfly populations are discussed.

DNA barcoding and molecular identification of field-collected Culicoides larvae in the Niayes area of Senegal.

BACKGROUND: Biting midge species of the genus Culicoides Latreille (Diptera: Ceratopogonidae) comprise more than 1300 species distributed worldwide. Several species of Culicoides are vectors of various viruses that can affect animals, like the African horse sickness virus (AHSV), known to be endemic in sub-Saharan Africa. The ecological and veterinary interest of Culicoides emphasizes the need for rapid and reliable identification of vector species. However, morphology-based identification has limitations and warrants integration of molecular data. DNA barcoding based on the mitochondrial gene cytochrome c oxidase subunit 1 (cox1) is used as a rapid and authentic tool for species identification in a wide variety of animal taxa across the globe. In this study, our objectives were as follows: (i) establish a reference DNA barcode for Afrotropical Culicoides species; (ii) assess the accuracy of cox1 in identifying Afrotropical Culicoides species; and (iii) test the applicability of DNA barcoding for species identification on a large number of samples of Culicoides larvae from the Niayes area of Senegal, West Africa. RESULTS: A database of 230 cox1 sequences belonging to 42 Afrotropical Culicoides species was found to be reliable for species-level assignments, which enabled us to identify cox1 sequences of Culicoides larvae from the Niayes area of Senegal. Of the 933 cox1 sequences of Culicoides larvae analyzed, 906 were correctly identified by their barcode sequences corresponding to eight species of Culicoides. A total of 1131 cox1 sequences of adult and larval Culicoides were analyzed, and a hierarchical increase in mean divergence was observed according to two taxonomic levels: within species (mean = 1.92%, SE = 0.00), and within genus (mean = 17.82%, SE = 0.00). CONCLUSIONS: Our study proves the efficiency of DNA barcoding for studying Culicoides larval diversity in field samples. Such a diagnostic tool offers great opportunities for investigating Culicoides immature stages ecology and biology, a prerequisite for the implementation of eco-epidemiological studies to better control AHSV in the Niayes region of Senegal, and more generally in sub-Saharan Africa.

Possible over-wintering of bluetongue virus in Culicoides populations in the Onderstepoort area, Gauteng, South Africa.

Several studies have demonstrated the ability of certain viruses to overwinter in arthropod vectors. The over-wintering mechanism of bluetongue virus (BTV) is unknown. One hypothesis is over-wintering within adult Culicoides midges (Diptera; Ceratopogonidae) that survive mild winters where temperatures seldom drop below 10 °C. The reduced activity of midges and the absence of outbreaks during winter may create the impression that the virus has disappeared from an area. Light traps were used in close association with horses to collect Culicoides midges from July 2010 to September 2011 in the Onderstepoort area, in Gauteng Province, South Africa. More than 500 000 Culicoides midges were collected from 88 collections and sorted to species level, revealing 26 different Culicoides species. Culicoides midges were present throughout the 15 month study. Nine Culicoides species potentially capable of transmitting BTV were present during the winter months. Midges were screened for the presence of BTV ribonucleic acid (RNA) with the aid of a real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay. In total 91.2% of midge pools tested positive for BTV RNA. PCR results were compared with previous virus isolation results (VI) that demonstrated the presence of viruses in summer and autumn months. The results indicate that BTV-infected Culicoides vectors are present throughout the year in the study area. Viral RNA-positive midges were also found throughout the year with VI positive midge pools only in summer and early autumn. Midges that survive mild winter temperatures could therefore harbour BTV but with a decreased vector capacity. When the population size, biting rate and viral replication decrease, it could stop BTV transmission. Over-wintering of BTV in the Onderstepoort region could therefore result in re-emergence because of increased vector activity rather than reintroduction from outside the region.

Culicoides species composition and environmental factors influencing African horse sickness distribution at three sites in Namibia.

African horse sickness (AHS) is one of the most lethal infectious, non-contagious, vector-borne disease of equids. The causative agent, African horse sickness virus (AHSV) is transmitted via Culicoides midges (Diptera: Ceratopogonidae). AHS is endemic to Namibia but detailed studies of Culicoides communities and influencing environmental parameters are limited. This study aims to determine the Culicoides species composition at three different sites and to assess environmental parameters influencing the geographical distribution of AHS in Namibia. Weekly collections of Culicoides were made during the AHS peak season from January to May for 2013 and 2014 using the Onderstepoort 220V UV-light trap. Out of 397 collections made, 124 collections (3287 Culicoides) were analysed for AHSV presence with RT-qPCR. A total of 295 collections were analysed for total Culicoides (all collected Culicoides individuals) and in 75% of these collections the Culicoides were identified to species level. C. imicola was the dominant species with proportional representation of 29.9%. C. subschultzei, C. exspectator and C. ravus each contribute more than 10% to the species composition. The lowest number of Culicoides was collected at Aus 9980, a total of 21819 at Windhoek and the highest number at Okahandja 47343. AHSV was present at all three sites during 2013 but only in Windhoek and Okahandja during 2014. Multivariate analyses of data from the two year survey indicate the environmental parameters in order of importance for the distribution of AHS in Namibia as precipitation>temperature>clay>relative humidity>NDVI. The implication of these findings is that any precipitation event increases Culicoides numbers significantly. Together with these results the high number of species found of which little is known regarding their vector competence, add to the complexity of the distribution of AHS in Namibia.

DNA barcoding and surveillance sampling strategies for Culicoides biting midges (Diptera: Ceratopogonidae) in southern India.

BACKGROUND: Culicoides spp. biting midges transmit bluetongue virus (BTV), the aetiological agent of bluetongue (BT), an economically important disease of ruminants. In southern India, hyperendemic outbreaks of BT exert high cost to subsistence farmers in the region, impacting on sheep production. Effective Culicoides spp. monitoring methods coupled with accurate species identification can accelerate responses for minimising BT outbreaks. Here, we assessed the utility of sampling methods and DNA barcoding for detection and identification of Culicoides spp. in southern India, in order to provide an informed basis for future monitoring of their populations in the region. METHODS: Culicoides spp. collected from Tamil Nadu and Karnataka were used to construct a framework for future morphological identification in surveillance, based on sequence comparison of the DNA barcode region of the mitochondrial cytochrome c oxidase I (COI) gene and achieving quality standards defined by the Barcode of Life initiative. Pairwise catches of Culicoides spp. were compared in diversity and abundance between green (570 nm) and ultraviolet (UV) (390 nm) light emitting diode (LED) suction traps at a single site in Chennai, Tamil Nadu over 20 nights of sampling in November 2013. RESULTS: DNA barcode sequences of Culicoides spp. were mostly congruent both with existing DNA barcode data from other countries and with morphological identification of major vector species. However, sequence differences symptomatic of cryptic species diversity were present in some groups which require further investigation. While the diversity of species collected by the UV LED Center for Disease Control (CDC) trap did not significantly vary from that collected by the green LED CDC trap, the UV CDC significantly outperformed the green LED CDC trap with regard to the number of Culicoides individuals collected. CONCLUSIONS: Morphological identification of the majority of potential vector species of Culicoides spp. samples within southern India appears relatively robust; however, potential cryptic species diversity was present in some groups requiring further investigation. The UV LED CDC trap is recommended for surveillance of Culicoides in southern India.

The occurrence of Culicoides species, the vectors of arboviruses, at selected trap sites in Zimbabwe.

A study of the distribution of Culicoides species was conducted by establishing 12 light trap sites over five rainy seasons between 1998 and 2003 covering all the geo-climatic natural regions of Zimbabwe. In total, 279 919 specimens of Culicoides were trapped over a total of 163 trapping nights. The highest median counts of Culicoides per trapping night were recorded in natural region III, which has climatic conditions conducive to the successful development of the larvae. Culicoides imicola, the major vector of bluetongue and African horse sickness viruses in Africa, was found to be the most abundant species (80.4%), followed by Culicoides enderleini (5.9%) and Culicoides milnei (5.2%). This study identified 10 species of Culicoides that had not been previously described in Zimbabwe, including Culicoides loxodontis and Culicoides miombo, which are members of the C. imicola complex. A total of 23 994 Culicoides midges were collected from five trap sites in Harare, Zimbabwe, with the dominant species, C. imicola, representing 91.6% of the total collection. Seventeen arboviruses were isolated from these midges, 15 of which were bluetongue virus. The predominant bluetongue virus serotype was serotype 11, followed by serotypes 1, 8, 12 and 15. Bluetongue virus serotypes 1, 2, 8, 10, 12, 15, 16 and 18, detected in this study, had not been previously reported in Zimbabwe.

How do species, population and active ingredient influence insecticide susceptibility in Culicoides biting midges (Diptera: Ceratopogonidae) of veterinary importance?

BACKGROUND: Culicoides biting midges are biological vectors of internationally important arboviruses of livestock and equines. Insecticides are often employed against Culicoides as a part of vector control measures, but systematic assessments of their efficacy have rarely been attempted. The objective of the present study is to determine baseline susceptibility of multiple Culicoides vector species and populations in Europe and Africa to the most commonly used insecticide active ingredients. Six active ingredients are tested: three that are based on synthetic pyrethroids (alpha-cypermethrin, deltamethrin and permethrin) and three on organophosphates (phoxim, diazinon and chlorpyrifos-methyl). METHODS: Susceptibility tests were conducted on 29,064 field-collected individuals of Culicoides obsoletus Meigen, Culicoides imicola Kieffer and a laboratory-reared Culicoides nubeculosus Meigen strain using a modified World Health Organization assay. Populations of Culicoides were tested from seven locations in four different countries (France, Spain, Senegal and South Africa) and at least four concentrations of laboratory grade active ingredients were assessed for each population. RESULTS: The study revealed that insecticide susceptibility varied at both a species and population level, but that broad conclusions could be drawn regarding the efficacy of active ingredients. Synthetic pyrethroid insecticides were found to inflict greater mortality than organophosphate active ingredients and the colony strain of C. nubeculosus was significantly more susceptible than field populations. Among the synthetic pyrethroids, deltamethrin was found to be the most toxic active ingredient for all species and populations. CONCLUSIONS: The data presented represent the first parallel and systematic assessment of Culicoides insecticide susceptibility across several countries. As such, they are an important baseline reference to monitor the susceptibility status of Culicoides to current insecticides and also to assess the toxicity of new active ingredients with practical implications for vector control strategies.

Assessment of the repellent effect of citronella and lemon eucalyptus oil against South African Culicoides species.

The use of insect repellents to reduce the attack rate of Culicoides species (Diptera:Ceratopogonidae) should form part of an integrated control programme to combat African horse sickness and other diseases transmitted by these blood-feeding midges. In the present study the repellent effects of a commercially available mosquito repellent, a combination of citronella and lemon eucalyptus oils, on Culicoides midges was determined. The number of midges collected with two 220 V Onderstepoort traps fitted with 8 W 23 cm white light tubes and baited with peel-stick patches, each containing 40 mg of active ingredient, was compared with that of two unbaited traps. Two trials were conducted and in each trial the four traps were rotated in two replicates of a 4 x 4 randomised Latin square design. Although more midges were collected in the baited traps, the mean number in the baited and unbaited traps was not significantly different. This mosquito repellent did not influence either the species composition or the physiological groups of Culicoides imicola Kieffer. The higher mean numbers in the baited traps, although not statistically significant, may indicate that this mosquito repellent might even attract Culicoides midges under certain conditions.

Culicoides species abundance and potential over-wintering of African horse sickness virus in the Onderstepoort area, Gauteng, South Africa.

In South Africa, outbreaks of African horse sickness (AHS) occur in summer; no cases are reported in winter, from July to September. The AHS virus (AHSV) is transmitted almost exclusively by Culicoides midges (Diptera: Ceratopogonidae), of which Culicoides imicola is considered to be the most important vector. The over-wintering mechanism of AHSV is unknown. In this study, more than 500 000 Culicoides midges belonging to at least 26 species were collected in 88 light traps at weekly intervals between July 2010 and September 2011 near horses in the Onderstepoort area of South Africa. The dominant species was C. imicola. Despite relatively low temperatures and frost, at least 17 species, including C. imicola, were collected throughout winter (June-August). Although the mean number of midges per night fell from > 50 000 (March) to < 100 (July and August), no midge-free periods were found. This study, using virus isolation on cell cultures and a reverse transcriptase polymerase chain reaction (RT-PCR) assay, confirmed low infection prevalence in field midges and that the detection of virus correlated to high numbers. Although no virus was detected during this winter period, continuous adult activity indicated that transmission can potentially occur. The absence of AHSV in the midges during winter can be ascribed to the relatively low numbers collected coupled to low infection prevalence, low virus replication rates and low virus titres in the potentially infected midges. Cases of AHS in susceptible animals are likely to start as soon as Culicoides populations reach a critical level.

Environmental drivers of Culicoides phenology: how important is species-specific variation when determining disease policy?

Since 2006, arboviruses transmitted by Culicoides biting midges (Diptera: Ceratopogonidae) have caused significant disruption to ruminant production in northern Europe. The most serious incursions involved strains of bluetongue virus (BTV), which cause bluetongue (BT) disease. To control spread of BTV, movement of susceptible livestock is restricted with economic and animal welfare impacts. The timing of BTV transmission in temperate regions is partly determined by the seasonal presence of adult Culicoides females. Legislative measures therefore allow for the relaxation of ruminant movement restrictions during winter, when nightly light-suction trap catches of Culicoides fall below a threshold (the 'seasonally vector free period': SVFP). We analysed five years of time-series surveillance data from light-suction trapping in the UK to investigate whether significant inter-specific and yearly variation in adult phenology exists, and whether the SVFP is predictable from environmental factors. Because female vector Culicoides are not easily morphologically separated, inter-specific comparisons in phenology were drawn from male populations. We demonstrate significant inter-specific differences in Culicoides adult phenology with the season of Culicoides scoticus approximately eight weeks shorter than Culicoides obsoletus. Species-specific differences in the length of the SVFP were related to host density and local variation in landscape habitat. When the Avaritia Culicoides females were modelled as a group (as utilised in the SFVP), we were unable to detect links between environmental drivers and phenological metrics. We conclude that the current treatment of Avaritia Culicoides as a single group inhibits understanding of environmentally-driven spatial variation in species phenology and hinders the development of models for predicting the SVFP from environmental factors. Culicoides surveillance methods should be adapted to focus on concentrated assessments of species-specific abundance during the start and end of seasonal activity in temperate regions to facilitate refinement of ruminant movement restrictions thereby reducing the impact of Culicoides-borne arboviruses.

The effect of anthropogenic activity on the occurrence of Culicoides species in the South-Western Khomas Region, Namibia.

Certain species of midges in the genus Culicoides (Diptera: Ceratopogonidae) are vectors of several serious orbiviral (Reoviridae) diseases, one of which, African horse sickness (AHS), was reported in the South-Western area of Khomas Region, Namibia, where it had been believed to be absent. Culicoides imicola, AHS principal vector, was collected in several farms in the area during the winter of 2009. The objective of this study was to determine whether Culicoides midges, especially C. imicola, were favoured at anthropogenic impacted/homestead sites in the arid Khomas Region, where they were not expected to occur under natural, veld conditions. The natural 'background' Culicoides communities where determined from collections made at veld sites, which were then compared to corresponding collections made at homestead sites. Altogether, 10,178 Culicoides midges were collected at homesteads and were then compared to 1,733 individuals collected at veld sites. Culicoides midge numbers were likely boosted in anthropogenic impacted areas/homesteads. This was also the case for the Culicoides species that are vector of AHS. This study indicated the significance of human settlement in the Khomas Region in terms of Culicoides midge abundance and distribution and showed the implications that this may have on the transmission of Culicoides-vectored diseases.

First record of Culicoides oxystoma Kieffer and diversity of species within the Schultzei group of Culicoides Latreille (Diptera: Ceratopogonidae) biting midges in Senegal.

The Schultzei group of Culicoides Latreille (Diptera: Ceratopogonidae) is distributed throughout Africa to northern Asia and Australasia and includes several potential vector species of livestock pathogens. The taxonomy of the species belonging to this species group is confounded by the wide geographical distribution and morphological variation exhibited by many species. In this work, morphological and molecular approaches were combined to assess the taxonomic validity of the species and morphological variants of the Schultzei group found in Senegal by comparing their genetic diversity with that of specimens from other geographical regions. The species list for Senegal was updated with four species: Culicoides kingi, C. oxystoma, C. enderleini and C. nevilli being recorded. This is the first record of C. oxystoma from Africa south of Sahara, and its genetic relationship with samples from Israel, Japan and Australia is presented. This work provides a basis for ecological studies of the seasonal and spatial dynamics of species of this species group that will contribute to better understanding of the epidemiology of the viruses they transmit.

The effect of high frequency sound on Culicoides numbers collected with suction light traps.

Culicoides midges (Diptera: Ceratopogonidae), are involved in the transmission of various pathogens that cause important diseases of livestock worldwide. The use of insect repellents to reduce the attack rate of these insects on livestock could play an important role as part of an integrated control programme against diseases transmitted by these midges. The objective of this study was to determine whether high frequency sound has any repellent effect on Culicoides midges. The number of midges collected with 220 V Onderstepoort white light traps fitted with electronic mosquito repellents (EMRs), emitting 5-20 KHz multi-frequency sound waves, was compared with that of two untreated traps. Treatments were rotated in two replicates of a 4 x 4 randomised Latin square design. Although fewer midges were collected in the two traps fitted with EMRs, the average number collected over eight consecutive nights was not significantly different. The EMRs also had no influence on any of the physiological groups of Culicoides imicola Kieffer or the species composition of the Culicoides population as determined with light traps. The results indicate that high frequency sound has no repellent effect on Culicoides midges. There is therefore no evidence to support their promotion or use in the protection of animals against pathogens transmitted by Culicoides midges.