Several areas of the bionomics of Culicoides in Virginia were given preliminary investigation. These included surveys for larval habitats, colonization attempts with Culicoides guttipennis (Coq.), host preference studies and adult activity studies. Breeding sites of 22 species of Culicoides were located and briefly described. Samples of mud, debris, etc. from suspected breeding sites were brought into the laboratory and maintained at 70°F in food containers with screened tops. Most species were found to prefer a specific type of habitat (i. e. tree-holes, polluted areas, etc.) but exceptions were occasionally observed. Colonization attempts with C. guttipennis have resulted in the successful establishment of this species in the laboratory. The colony has been maintained for 12-15 generations and is currently producing over 1,000 adults per day and an excess of eggs. Techniques developed for larval and adult maintenance have proved to be highly successful in recent months. Adults were held in a constant temperature cabinet at 80°F ± 2° and 85% relative humidity ± 10% in semi-darkness (less than 1 ft-c ). Eggs were collected on moist filter paper exposed in shell vials. Fly eggs hatched in 3 days and the young larvae were introduced into aquaria containing leaf mold from hardwood forests and distilled water. Larvae pupated in about 12 days and adults emerged about 3 days later. The adult diet consisted of rabbit blood, raisins, sugar cubes, honey water and a 6:6:1:13 mixture of powdered milk, sugar, egg solids and water administered in capillary tubes. In host preference studies 15 species of Culicoides were collected on 1 or more of 14 different hosts used in animal-baited traps. Animals were restrained and exposed on a small platform. After 15 min the animals were covered with a collecting cage. Flies were aspirated from the traps and killed in 70% ethanol. Even though some preferences possibly were shown by some species, more data will be needed before definite conclusions can be drawn. In adult activity studies, most species appeared to be more abundant from 9 PM to 3 AM as indicated by light traps. This was especially noted in C. stellifer, C. haematopotus and C. venustus. The activity of C. haematopotus was fairly constant up to 3 AM and then decreased. C. stellifer and C. venustus activity decreased as the night progressed. / Doctor of Philosophy

An artificial larval medium for colonized Culicoides guttipennis was developed. Microorganisms in decaying leaf matter were present in the original laboratory diet. These were believed to be the main food of C. guttipennis larvae. This was verified by sterilizing the leaf matter to kill the organisms present. Larvae did not develop beyond the first instar in this medium, but adults were produced from non-sterilized leaf matter. The objective was to develop cultures of various microorganisms and find one that would be best for rearing C. guttipennis larvae in producing more adults. A simple hay-infusion medium inoculated with stump-hole microorganisms was found more effective than all other cultures screened in rearing the larvae. This culture consisted of stump-hole microorganisms inoculated fresh each week from stored stumphole contents. It was compared with the same culture containing stumphole microorganisms completely adapted to a laboratory environment. The former was better in rearing C. guttipennis larvae. A final experiment evaluated the incorporation of vermiculite as an artificial substrate. This proved successful in rearing larvae and producing large numbers of adults. This culture was superior over the larval rearing medium of decaying leaf matter. Development of a successful artificial larval medium for colonized C. guttipennis will aid in establishing other laboratory colonies of related Culicoides species. / Master of Science

Les maladies animales vectorielles sont devenues depuis 2000 une préoccupation majeure pour la France et certains pays de l'Union Européenne du fait de l'apparition, la persistance et l'expansion de virus pathogènes en provenance de régions du Sud. La fièvre catarrhale du mouton (FCM) est un bon exemple de la progression en zone tempérée d'arboviroses tropicales sous l'effet, semble-t-il, de l'extension de son vecteur principal sur le littoral méditerranéen européen (Corse, îles Baléares, Catalogne, Italie, Sardaigne, Sicile, Var).<br />L'objectif de cette thèse est à la fois d'étudier les outils méthodologiques et diagnostiques disponibles pour évaluer les risques liés à la FCM et d'en développer d'autres afin d'améliorer la surveillance de cette maladie en France.<br />Une première étude a constitué en l'évaluation des performances épidémiologiques du test sérologique (ELISA de compétition) utilisé pour le diagnostic et la surveillance de la FCM en Corse et sur le continent français. Une analyse du plan de surveillance de la circulation virale dans le contexte particulier corse a également été conduite. Elle a permis notamment d'identifier les points critiques de ce dispositif. <br />Suite à la détection de la présence de Culicoides imicola, vecteur biologique impliqué dans la circulation du virus de la FCM (BTV) sur le pourtour méditerranéen, dans la vallée de l'Argens (département du Var), le besoin de développer de nouveaux outils pour surveiller et évaluer le risque de transmission de ce virus dans des zones d'implantation récente est devenu nécessaire. Une étude entomologique a ainsi été mise en place dans la vallée de l'Argens afin i) de confirmer l'installation de C. imicola dans cette zone, ii) de suivre la dynamique de cette population sur une saison d'activité, iii) de collecter des données entomologiques pour estimer le potentiel de transmission de cette population de vecteurs. Un modèle stochastique de capacité vectorielle regroupant des données spécifiques des populations locales de C. imicola et des données publiées dans la littérature est ainsi proposé. Une étude de la compétence de C. imicola vis-à-vis du sérotype 9 de la FCM a, de plus, été initiée.<br />La discussion générale s'attarde sur les points qui semblent importants pour améliorer le dispositif de surveillance et avancer vers la mise en place d'un système d'alerte précoce. Elle ouvre également de nouvelles perspectives pour prolonger l'approche intégrative proposée en incluant notamment des travaux de géomatique initiés sur C. imicola. Elle pose, de plus, la question d'un passage à une échelle d'étude plus large. Les questions de recherche que pose l'épizootie récente de FCM dans le Nord de l'Europe sont finalement examinées.

Dans un contexte d’émergence ou de réémergence des maladies à transmission vectorielle, certaines espèces de Culicoides (Diptera : Ceratopogonidae) sont impliquées dans la transmission de certains virus (Reoviridae : Orbivirus) et de nématodes (Onchocercidae : Mansonella) en région Afrotropicale. Cependant, le schéma systématique et taxonomique ainsi que la bio-écologie des espèces d’intérêt vétérinaire restent à explorer. Ce travail de taxonomie intégrative a pour objectifs de réaliser (i) une révision systématique et taxonomique des espèces appartenant à des sous-genres et des groupes d’intérêt à partir de phylogénie moléculaire multi-marqueur et de délimitation d’espèces, et (ii) de développer des outils moléculaires pour l’étude de la bio-écologie des espèces d’intérêt vétérinaire et de la dynamique de leurs populations d’immatures. Nos résultats mettent en évidence (i) la présence de trois clades monophylétiques le groupe d’espèces Imicola, le groupe d’espèces Milnei et le sous-genre Remmia, (ii) une nouvelle espèce pour la science nommée C. sp. #22 et affiliée au sous-genre Avaritia, groupe Imicola, (iii) la présence d’une nouvelle espèce non-décrite nommée C. sp. #54 appartenant au groupe d’espèces Dasyops, sous-genre Avaritia, (iii) d’affilier les groupes d’espèces Similis et Neavei au sous-genre Synhelea et, (iv) de poser l’hypothèse d’une présence d’espèces cryptiques au sein de C. oxystoma (sous-genre Remmia). D’un point de vue bio-écologique, le travail réalisé combinant suivi entomologique et identification moléculaire avec une librairie de séquences barcodes a permis de décrire le comportement trophique de C. imicola, C. kingi et C. oxystoma ainsi que leurs habitats larvaires dans des environnements équins de la zone des Niayes au Sénégal. L’ensemble de ce travail permet de compléter le corpus de connaissances sur le genre Culicoides en région Afrotropicale afin d’améliorer la compréhension de l’épidémiologie des pathogènes transmis et proposer des pistes de recherches pour mieux contrôler les populations immatures et adultes des espèces vectrices afin de mieux anticiper et prévenir des évènements sanitaires. / In a context of emergence or re-emergence of vector-borne diseases, certain species of Culicoides (Diptera: Ceratopogonidae) are involved in the transmission of certain viruses (Reoviridae: Orbivirus) and nematodes (Onchocercidae: Mansonella) in the Afrotropical region. However, the systematic and taxonomic schemes as well as the bio-ecology of species of veterinary interest remain to be explored. This work of integrative taxonomy aims to achieve (i) a systematic and taxonomic revision of species belonging to subgenera and groups of veterinary interest using a multi-marker molecular phylogeny and species delineation, and (ii) to develop molecular tools for studying the bioecology of species of veterinary interest and dynamics of their immature populations. Our results show (i) the presence of three monophyletic clades, the Imicola group, the Milnei group and the subgenus Remmia, (ii) a new species for science named C. sp. # 22 and affiliated into the subgenus Avaritia, Imicola group, (iii) the presence of a new undescribed species named C. sp. # 54 belonging to the Dasyops group, subgenus Avaritia, (iii) affiliating the Similis and Neavei species groups to the subgenus Synhelea, and (iv) cryptic species within C. oxystoma (subgenus Remmia). From a bioecological point of view, this work combining entomological follow-up and molecular identification with a library of barcode sequences allowed to describe the trophic behavior of C. imicola, C. kingi and C. oxystoma as well as their larval habitats in equine environments of the Niayes area in Senegal. This work completes the corpus of knowledge about the genus Culicoides in the Afrotropical region to improve our knowledge on the epidemiology of the transmitted pathogens and to propose research tracks to better control the immature and adult populations of the vector species in order to better anticipate and prevent Culicoides-borne diseases outbreaks.

La maladie épizootique hémorragique (EHD) est une arbovirose inscrite sur la liste de l'OIE (organisation mondiale de la santé), qui affecte aussi bien les ruminants sauvages (essentiellement les cervidés) que les ruminants domestiques (les bovins) par la morsure de petits moucherons hématophages, les Culicoïdes (Diptera: Ceratopogonidae). Au début du 21ème siècle, cette maladie a émergé au Maghreb et au Moyen Orient (Turquie, Israël et Jordanie), générant de lourdes pertes économiques pour les éleveurs.L'émergence inattendue de l'EHDV en Tunisie en 2006, simultanément avec l’observation de foyers d'EHDV-6 au Maroc et en Algérie, a suscité une vive inquiétude dans le monde agricole tunisien, qui avait déjà eu à faire face à la survenue d’épizooties d'autres orbivirus : la peste équine africaine en 1960 et de la fièvre catarrhale ovine en 1999 donnant l’exemple qu'une maladie, réputée exotique, puisse émerger et devenir endémique (cas pour la fièvre catarrhale ovine).Ce projet a donc été réalisé pour améliorer les connaissances sur le virus de l’EHDV responsable de l'infection en Tunisie, en 2006, qui entrainait l’apparition de signes cliniques semblables à ceux du BTV.Dans un premier temps, nous avons disposé d'un échantillonnage de différents prélèvements de bovins, Culicoïdes de type imicola collectés en 2006 et conservés à la sérothéque de l'IRVT. Nous y avons donc recherché le génome de l’EHDV par RT-PCR afin de le caractériser et d'isoler le virus. Les résultats ont démontré que l'EHDV-6 était bien le sérotype qui circulait en Tunisie en 2006. Cette partie du travail a fait l'objet d'une publication dans Acta Veterinaria Hungarica.Le deuxième travail exposé s'est intéressé à la recherche d'une présence éventuelle de l’EHDV-6 en Tunisie avant et après l'épidémie de 2006 dans deux espèces animales : les bovins et les dromadaires. Pour cela, nous avons recherché les anticorps ou le génome viral à partir de prélèvements de terrain réalisés de 2000 à 2014 sur des bovins et des dromadaires. A l'issue de cette étude, nous avons détecté une possible circulation à très bas bruit de l'EHDV-6 chez les bovins. Les résultats obtenus sont discutés et ont fait l'objet d'une publication en cours de publication dans le journal Véterinaria Italiana.Concernant l’étude réalisée chez le dromadaire, espèce sensible au BTV, nous voulions évaluer son rôle de réservoir potentiel pour le virus de l’EHD. L’ensemble des résultats sérologiques et virologiques de notre étude nous indique que cette espèce ne semble pas jouer un rôle dans l’épidémiologie de l’EHD.Enfin, parallèlement à ces recherches sur le virus de l’EHD, nous avons mené une enquête sur la présence du virus de la Bluetongue à partir des échantillons de dromadaires et de bovins tunisiens. Les résultats ont été discutés. L'ensemble de ces études contribue à une meilleure connaissance de l'EHDV-6 présent en Tunisie, et permet de rendre compte des espèces potentiellement réservoirs. Certains travaux présentés pourraient être poursuivis pour évaluer le rôle du dromadaire comme réservoir d’Orbivirus et mieux déterminer les espèces de l'inventaire faunique des Culicoïdes impliquées dans la transmission des Orbivirus. / The epizootic hemorrhagic disease (EHD) is an arthropod-borne virus that is on the OIE’s list (World Animal Health Organisation, formerly Office international des épizooties), this disease is mainly transmitted to wild (mainly deer) as well as domestic (primarily cattle) ruminants, by the bites of minute size midges, the culicoides (Diptera: Ceratopogonidae) also known as biting midges. In the beginning of the 21st century, EHD was emerged in Maghreb (North Africa) and in the Middle East (Turkey, Israel, Jordan), causing severe losses for the farmers and ranchers.The unexpected emergency of EHDV in Tunisia in 2006, simultaneously with the observation of EHDV-6 cases in Morocco and Algeria, has aroused great concern in the Tunisian agricultural sector, which had already to face the occurrence of other animal diseases orbivirus: African horse sickness in 1966 and bluetongue in 1999 giving examples of the possibility that deemed exotic disease could emerge and become endemic (case bluetongue).This project was carried out to raise the knowledge on the EHDV virus causing the infection in Tunisia in 2006, which led to the appearance of clinical signs similar to those of BTV.First, we prepared a sampling of various samples of cattle, Culicoides type imicola collected in 2006 and stored at the serum bank of IRVT. So, we searched therefore the genome of the EHDV by RT-PCR in order to characterize and isolate the virus. Results showed that EHDV-6 was actually the serotype circulating in Tunisia in 2006. This part of the job was published in Acta Veterinaria Hungarica.The second working paper concerned the potential presence of the EHDV-6 in Tunisia before and after the epidemic of 2006 in two animal species: cattle and camels. For this we looked for antibodies or viral genome from field samples collected from 2000 to 2014 cattle and camels. Following this study, we detected a possible circulation of EHDV-6 at a very low level of intensity among the cattle. The found results were discussed and made the subject of a publication to be in the newspaper Veterinaria iItaliana.Regarding the study on the dromedary species sensitive to BTV, we wanted to examine its potential role as a reservoir species for EHD virus. All serological and virological results of our study indicate that this species does not seem to play a role in the epidemiology of EHD.Finally, alongside these researches on EHD virus, we have investigated the presence of Bluetongue Virus in Tunisian samples from camels and cattle. The results were discussed.All these studies contribute to a better knowledge of EHDV-6 present in Tunisia and allows taking into account some species that are potentially reservoir. Some presented researches could be pursued to assess the role of the camel as a reservoir for Orbivirus and better identify species of the faunal inventory of Culicoides involved in transmission of orbivirus.

La découverte du rôle des insectes en tant que vecteurs de pathogènes, établi depuis plus d'un siècle, a été l'élément moteur de la discipline « entomologie médicale et vétérinaire ». Malgré le succès de nombreuses campagnes de prévention et de programmes de lutte, nous assistons depuis une trentaine d'années à l'émergence et à la recrudescence de maladies à transmission vectorielle. Le virus de la fièvre catarrhale ovine (FCO) (Reoviridae : Orbivirus) est un très bon exemple de virus émergent en Europe dont les mécanismes de transmission sont encore peu connus dans cette région. Ce virus est transmis par des moucherons hématophages du genre Culicoides (Diptera : Ceratopogonidae) aux ruminants sauvages et domestiques. En Europe, la FCO a été pendant longtemps considérée comme une maladie exotique. À partir de 1998, plusieurs incursions apparaissent dans l'ouest du bassin méditerranéen en lien avec la remontée vers le nord de populations de Culicoides imicola, le principal vecteur afrotropical. À partir d'août 2006, l'apparition et la transmission du sérotype 8 dans le nord de l'Europe, dans des zones où C. imicola est absent, révèle l'importance des espèces autochtones et la nécessité de comprendre leur rôle vecteur. Ce travail s'intéresse aux mécanismes de transmission du virus de la FCO en Europe non méditerranéenne, en i) présentant un état de l'art de la biologie et l'écologie des Culicoides adultes, ii) en évaluant les conditions possibles d'utilisation de pièges pour estimer le taux de piqûre et iii) en décrivant les comportements trophiques pour les espèces d'intérêt vétérinaire. / The discovery of insects as pathogens vectors was established for over a century and was the driving force behind the discipline “medical and veterinary entomology”. Despite the success of some prevention and control program campaigns, the emergence and spread of vector-borne diseases occurred dramatically during this last thirty years. Bluetongue virus (BTV) (Reoviridae: Orbivirus) is a good example of emerging virus in Europe, with a little understanding of the epidemiology of this disease. This virus is transmitted by blood-sucking midges of the genus Culicoides (Diptera: Ceratopogonidae) to wild and domestic ruminants. In Europe, BT was considered an exotic disease. In 1998, several incursions appeared in the western Mediterranean Basin in line with the northward progression of C. imicola populations, the main afrotropical vector. From August 2006, the emergence and transmission of serotype 8 in northern Europe, in areas where C. imicola is absent, revealed the importance of autochthonous species and the urgent need to understand their role as vector. This work gives new insights into the understanding of BTV transmission in northern Europe: i) presenting a state of the art review of the biology and ecology of Culicoides adults, ii) assessing different methods to study the biting rate and iii) highlighting trends in host-seeking behavior.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / Bluetongue (BT) is an infectious disease caused by bluetongue virus (BTV), which is transmitted by biting midges of the genus Culicoides, and affects domestic and wild ruminants, but its clinical manifestation is seen basically in sheep. Currently, 26 BTV serotypes are recognized worldwide. However, information about the disease in Brazil are limited, as only two BTV serotypes have been reported. Serological surveys reveal that Rio Grande do Sul (RS) has the lowest prevalence rates of infection among Brazilian states. This article describes the clinical, pathological and epidemiological aspects of 17 outbreaks of BT disease in sheep in the Central Region of RS state, Southern Brazil. Affected farms were visited for clinical examination, necropsy, sample collection and epidemiological investigation. The outbreaks were seasonal and occurred during the summer and fall. Of the 884 sheep in 17 small herds, 180 (20.4%) were affected. All ages of Texel and mixed breed sheep were affected. However, lambs (younger than one year) had higher morbidity than adult sheep. The most frequent clinical signs were anorexia, lethargy, loss of body condition, facial swelling mainly involving the lips, and greenish seromucous nasal discharge. Pulmonary edema, cardiac, skeletal muscle and esophageal striated muscle necrosis were the most prevalent findings. Erosive and ulcerative lesions in the upper gastrointestinal tract, and hemorrhage in the pulmonary artery also were common. The bluetongue virus (BTV) genome was detected by RT-PCR in blood, spleen, and lungs samples of 21 animals from 17 outbreaks. The virus involved in the outbreak 3 was isolated and shown to belong to serotype 17, for the first time reported in Brazil. Serology performed by agar gel immunodiffusion test (AGID) in 20 contact cattle showed seroconversion to BTV in 17 animals. In summary, our data support the BTV as the etiological agent of the outbreaks and indicate that the central region of RS is an area at risk for BT in sheep, a disease previously not recognized in the region. / Língua azul ou bluetongue (BT) é uma doença infecciosa causada pelo vírus da língua azul (BTV), que é transmitido por vetores hematófagos do gênero Culicoides e acomete ruminantes domésticos e selvagens, porém sua manifestação clínica é vista basicamente em ovinos. Atualmente, 26 sorotipos do BTV são conhecidos mundialmente. Entretanto, informações sobre a doença no Brasil são limitadas, com apenas dois sorotipos descritos. Pesquisas sorológicas revelam que o Rio Grande do Sul (RS) possui as menores taxas de prevalência de infecção entre os Estados brasileiros. O objetivo deste trabalho é descrever os aspectos epidemiológicos, clínicos e anatomopatológicos de 17 surtos de BT em ovinos na Região Central do RS. Para isso foram realizadas visitas às propriedades em que ocorreram os surtos para investigação epidemiológica e clínica, realização de necropsias e coleta de amostras. Os surtos foram sazonais e ocorreram durante o verão e outono. Em 17 pequenos rebanhos, de um total de 884 ovinos, 180 adoeceram (20,4%). Ovinos de todas as faixas etárias, da raça Texel e sem raça definida, foram acometidos. Entretanto, ovinos com menos de um ano de idade tiveram taxa de morbidade maior do que ovinos com um ano ou mais. Os sinais clínicos mais frequentes caracterizaram-se por anorexia, apatia, acentuada perda de peso, edema facial, envolvendo principalmente os lábios, e secreção nasal seromucosa e esverdeada. Edema pulmonar, necrose da musculatura cardíaca e esquelética e do músculo estriado do esôfago foram as lesões mais prevalentes. Erosões e úlceras no trato gastrointestinal superior e hemorragia na artéria pulmonar também foram comuns. O genoma do BTV foi detectado por meio da RT-PCR em amostras de sangue, baço e pulmão de 21 animais dos 17 surtos. O vírus envolvido no surto 3 foi isolado e pertence ao sorotipo 17, que pela primeira vez é descrito no Brasil. A sorologia foi realizada pelo método de imunodifusão em gel de ágar e demonstrou que 17 dos 20 bovinos que estavam em contato com os ovinos infectados soroconverteram ao BTV. Em síntese, nossos dados permitem concluir que o BTV é o agente causador dos surtos e indicam que a Região Central do RS é uma área de risco para a ocorrência de BT em ovinos, uma doença, até então, não reconhecida nessa região.

Doctor of Philosophy / Department of Entomology / Ludek Zurek / The larval stages of Culicoides sonorensis Wirth and Jones, a confirmed vector of bluetongue and epizootic hemorrhagic disease viruses affecting ruminants in North America, have been observed to occur typically in animal waste enhanced muds. In this dissertation, I studied the larval development (first instar to adult stage) and oviposition (four-choice assays) of C. sonorensis on sterilized mud (autoclaved) enriched with manure of different farm animal species (dairy cattle, beef cattle, sheep, goats, pigs, horses, white-tailed deer, and chicken). In addition, to determine why only some manure-polluted sites are colonized by C. sonorensis even when they are in close proximity to each other, I examined the moisture levels and microbial concentrations (mud) and physicochemical characteristics (standing water) of a manure-overflow pond site producing C. sonorensis and compared them to nearby cattle stock pond site(s) that produced different Culicoides species. Finally, as the first step in examining the role of microbiome in various physiological functions of C. sonorensis and other suspected/potential vector Culicoides species, I assessed the bacterial communities in field-collected adult females of C. sonorensis, C. crepuscularis, C. haematopotus, and C. stellifer (Illumina sequencing of 16S rRNA gene). In larval development experiments, the proportion of adults emerged and development time to adult stage varied with manure type and its concentration present in the substrate. Mud supplemented with chicken manure did not support C. sonorensis development, mud enriched with white-tailed deer manure poorly supported midge development, while C. sonorensis development in mud enhanced with manure of sheep, goats, beef cattle, dairy cattle, pigs, and horses varied. In oviposition experiments, colonized females preferred to deposit eggs on substrates without animal manure over substrates with animal manure. In subsequent studies, the manure-overflow pond site that produced mainly C. sonorensis contained significantly higher total aerobic culturable bacteria, pH, salinity, total dissolved solids, and conductivity levels than cattle stock pond sites that produced different Culicoides species. Finally, bacterial composition of field-collected C. sonorensis adult females comprised mainly of the phyla Proteobacteria and Firmicutes, while the majority of bacterial taxa identified from C. crepuscularis, C. haematopotus, and C. stellifer belonged to Proteobacteria. An unidentified bacterial genus (related to Tumebacillus), Propionibacterium, and Curvibacter were detected commonly across all four midge species. These results suggest that manure of several farm animal species can contribute to C. sonorensis development in the field. However, oviposition preferences remain uncertain, as colonized females appeared to show aversion to animal manure, which is in contradiction to the typical presence of C. sonorensis larvae in animal waste enhanced muds. Nonetheless, variations in microbial and/or physicochemical conditions in the larval habitats likely play a role in the differential emergence of C. sonorensis from various manure-polluted sites. Moreover, some bacterial taxa are associated commonly with C. sonorensis and other suspected/potential vector Culicoides species. Future studies are needed to examine oviposition preferences of field-collected females, life history traits of adults emerging from various manure-enriched substrates, developmental requirements of larvae, and the role of microbiome in various physiological functions of the host including vector competence for orbiviruses.

African horse sickness (AHS) was reported in the south-western Khomas Region, central Namibia (22° 24.063´ S, 17° 01.791´ E; 23° 32.617´ S, 15° 53.870´ E), contrary to expectations that the arid conditions in the area would limit its occurrence. This prompted investigation into the occurrence of AHS in horses, a possible reservoir animal, the Hartmann’s mountain zebra (Equus zebra. hartmannae) and the occurrence of the Culicoides midge vector (Diptera: Ceratopogonidae) of the disease in the area. Questionnaires were used to explore the geographic characteristics of the study area, the occurrence of an expected AHS virus reservoir animal, E. z. hartmannae and AHS in horses in the study area. According to the questionnaire, rainfall patterns seem to follow topography of the area, where the north-east formed the higher rainfall (420 mm/a) high-ground and the south-western formed the lower rainfall (120 mm/a) pediment zone in the south-west. Cases of AHS occurred mostly in mid-rainfall zones. E. z. hartmannae were present throughout the area. They migrated from the southwest towards the north-eastern high-grounds during droughts, presumably along ephemeral river beds. E. z. hartmannae were sampled for blood and tissues and analysed for evidence of African Horse Sickness Virus (AHSV) infection by indirect ELISA, RT-PCR and virus isolation techniques. All useable samples tested positive for anti-AHSV antibodies. Viral RNA was demonstrated in 26% of all the zebra sampled. No viable viruses were isolated from these samples, however further research is required, as difficult sampling conditions may have yielded false-negatives. From 6 July to 21 September 2009, Culicoides midges were collected during the dry winter season in suction UV-light traps installed at five selected sites along a rainfall gradient. In 38 collections, a total of 9091 Culicoides individuals, representing 25 species were collected. The dominance of the proven vector of AHSV, Culicoides imicola Kieffer, varied in dominance from 94% near Windhoek with high altitude and relatively higher annual rainfall, to 12% at the site situated farthest southwest, with the lowest altitude and annual rainfall. From what was observed of the occurrence of AHS in horses, E. z. hartmannae and the distribution and abundance of the AHSV vector (Culicoides spp.), it was concluded that AHS can be maintained in the south-western Khomas Region even in the lowest mean annual rainfall zones. / Thesis (MSc (Environmental Sciences))--North-West University, Potchefstroom Campus, 2012

African horse sickness (AHS) was reported in the south-western Khomas Region, central Namibia (22° 24.063´ S, 17° 01.791´ E; 23° 32.617´ S, 15° 53.870´ E), contrary to expectations that the arid conditions in the area would limit its occurrence. This prompted investigation into the occurrence of AHS in horses, a possible reservoir animal, the Hartmann’s mountain zebra (Equus zebra. hartmannae) and the occurrence of the Culicoides midge vector (Diptera: Ceratopogonidae) of the disease in the area. Questionnaires were used to explore the geographic characteristics of the study area, the occurrence of an expected AHS virus reservoir animal, E. z. hartmannae and AHS in horses in the study area. According to the questionnaire, rainfall patterns seem to follow topography of the area, where the north-east formed the higher rainfall (420 mm/a) high-ground and the south-western formed the lower rainfall (120 mm/a) pediment zone in the south-west. Cases of AHS occurred mostly in mid-rainfall zones. E. z. hartmannae were present throughout the area. They migrated from the southwest towards the north-eastern high-grounds during droughts, presumably along ephemeral river beds. E. z. hartmannae were sampled for blood and tissues and analysed for evidence of African Horse Sickness Virus (AHSV) infection by indirect ELISA, RT-PCR and virus isolation techniques. All useable samples tested positive for anti-AHSV antibodies. Viral RNA was demonstrated in 26% of all the zebra sampled. No viable viruses were isolated from these samples, however further research is required, as difficult sampling conditions may have yielded false-negatives. From 6 July to 21 September 2009, Culicoides midges were collected during the dry winter season in suction UV-light traps installed at five selected sites along a rainfall gradient. In 38 collections, a total of 9091 Culicoides individuals, representing 25 species were collected. The dominance of the proven vector of AHSV, Culicoides imicola Kieffer, varied in dominance from 94% near Windhoek with high altitude and relatively higher annual rainfall, to 12% at the site situated farthest southwest, with the lowest altitude and annual rainfall. From what was observed of the occurrence of AHS in horses, E. z. hartmannae and the distribution and abundance of the AHSV vector (Culicoides spp.), it was concluded that AHS can be maintained in the south-western Khomas Region even in the lowest mean annual rainfall zones. / Thesis (MSc (Environmental Sciences))--North-West University, Potchefstroom Campus, 2012