Dissertations / Theses on the topic 'Ceratopogonidae'

This study investigates aspects of the biology of C. impunctatus with a view to producing an optimised laboratory culture of this species. Artificial membrane feeding of C. impunctatus is shown to be enhanced by the addition of blood vestiges to the outer surface of the feeding apparatus. The effects of holding temperature, environment and conspecific density upon oogenesis in blood fed female midges are assessed through mortality, digestion of the blood meal and development of the eggbatch over time. Oviposition in C. impunctatus is investigated through choice and nochoice bioassays which show Sphagnum spp. Mosses to be highly effective in this regard. Juncus articulatuslacutiflorus infusions also significantly increase the number of eggs laid by females in comparison to oviposition substrates used in colonisation of other midge species. Preliminary studies are also carried out to provide a suitable larval medium for colonisation purposes. The possibility of future colonisation is discussed with reference both to those experiments carried out, and to those areas not yet addressed in the C. impunctatus lifecycle. Dispersal of C. impunctatus is also examined, both on a local scale and in terms of gene flow between UK populations. Capture, mark, recapture studies showed the marked population of C. impunctatus remained relatively close to the release site in the habitat used for the study. The effect of prevailing winds, however, was found to be highly important in terms of passive directional movements of individuals. Daily survival rates of parous females were calculated as very low in relation to other species of midge making this species an unlikely candidate for pathogen transfer in Scotland. Polymorphism in the molecular markers used to examine gene flow in C. impunctatus across the UK was minimal and precluded phylogenetic analysis. This result is interpreted in terms of both possible long-distance dispersal and the effects of rapid post-glacial popUlation expansion into the UK.

This study has investigated the nature of host derived cues used by parous female C. impunctatus in the location of blood meal hosts. Olfaction has been shown to be a vital part of the process. The responses of antennal receptors of females to a variety of host-derived volatile compounds were assessed using the EAG technique. High antennal sensitivity to olfactory cues was indicated by low thresholds for response and, taking into account the volatility of the compounds, a clear hierarchy of response was obtained in which octenol was the most stimulatory, followed by phenolics, lactic acid and finally ketones. Of particular note with regard to phenolic compounds was the finding that 3-derivatives (3-n-propylphenol & 3-methylphenol) were more stimulatory than their 4-derivatives (eg 4-methylphenol). Behavioural assays conducted within a Y-tube olfactometer showed that attraction to olfactory cues was dose dependent. Supra optimal doses which caused receptor saturation (as demonstrated in the EAG assay), also induced marked changes in behaviour with responses being either no different to random expectation, or the compound appearing to be repellent. Wind tunnel assays corroborated these findings for octenol and acetone when used in conjunction with 0.01% CO₂. Upwind flight was found to be maximal, and significantly different from responses to the CO₂ standard, at concentrations within the tunnel of 1.33x10^-8 g 1^-1 and 1.5x10^-6 g 1^-1, respectively. These concentrations are approximately those released by a bovid host for octenol and 1/1000x for acetone. Females exhibited a linear dose dependent response to increases in the concentration of CO₂ in the wind tunnel up to 0.09% whether gas was released as a plume or as a lightly turbulent cloud. However, at cloud concentrations above this, CNS mediated habituation is suggested to have arrested upwind flight.

Les Ceratopogonidae incluent 6180 espèces modernes et 274 espèces fossiles. Nous ajoutons douze nouvelles espèces : de la France (leptoconops daugeroni Choufani, Azar & Nel, 2011, leptoconops gravesi Choufani & Nel, 2014, leptoconops sp. Choufani & Nel, 2014, culicoides doyeni Choufani & Nel, 2014, devalquia brisaci Choufani & Nel, 2013, et metahelea roggeroi Choufani & Nel, 2013) du Liban (lebanoculicoides daheri Choufani & Nel, 2014, protoculicoides krzeminskii Choufani & Nel, 2014, archiaustroconops annae Choufani & Nel, 2014, archiaustroconops hammanensis Choufani & Nel, 2014 et archiaustroconops dominiakae Choufani & Nel, 2014) et de la Syrie (lebanoculicoides bloudani Choufani, 2015). La répartition mondiale des gisements contenant les fossiles et les habitudes alimentaires de la famille sont révisées. Une étude phylogénétique établie sur 91 caractères et 49 taxons représentant les cinq sous-familles et six tribus, remet en question la définition et la position de plusieurs genres
The Ceratopogonidae include 6180 modern species and 274 fossil species. We add twelve new species, from France (leptoconops daugeroni Choufani, Azar & Nel, 2011, leptoconops gravesi Choufani & Nel, 2014, leptoconops sp. Choufani & Nel, 2014, culicoides doyeni Choufani & Nel, 2014, devalquia brisaci Choufani & Nel, 2013, et metahelea roggeroi Choufani & Nel, 2013), Lebanon (lebanoculicoides daheri Choufani & Nel, 2014, protoculicoides krzeminskii Choufani & Nel, 2014, archiaustroconops annae Choufani & Nel, 2014, archiaustroconops hammanensis Choufani & Nel, 2014 et archiaustroconops dominiakae Choufani & Nel, 2014) and Syria (lebanoculicoides bloudani Choufani, 2015). The worlwide distribution of amber deposits containing fossils and the feeding habits of the family are reviewed. A phylogenetic study of 91 charcters comparing 49 taxa representingthe five subfamilies ans six tribes is established allowing questionning the definition and position of several genera

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The Ceratopogonidae family possesses four hematophagics genus: Forcipomyia, Leptoconops, Austroconops and Culicoides. The Culicoides are insects with 3mm in length, the female are vectors of helminths, protozoan and virus. The main arboviruses transmitted are Oropouche virus, endemic of amazon basin and Bluetongue Virus, which affects both wildlife and domestic livestocks. The convenience collections were done on Alvorada do Oeste, Buritis, Cacoal, Costa Marques, Espigão do Oeste, Guajará-Mirim, Pimenta Bueno, São Francisco Guaporé and Porto Velho municipalities, in Rondônia state through HP light traps between 18:00 and 06:00. Systematic collections were made in Porto Velho municipality on forest and pasture environments. Were collected 2,196 Culicoides (1,515 females and 681 males) distributed throughout 43 species and 27 morfotypes. The species most frequent were Culicoides insignis (n=1,594; 72.59%), C. foxi (n=76; 3.46%), C. hildebrandoi (n=53; 2.41%), C. leopoldoi (n=44; 2%), C. ocumarensis (n=34; 1.55%), C. pusillus (n=29; 1.32%) and C. glabrior (n=28; 1.28%). There were 37 news records of species in Rondônia and three for Brazil. The municipalities with greatest richness were Porto Velho (56), Cacoal (26) e Costa Marques (14). The municipalities with lowest richness were Espigão do Oeste (4), Buritis (3) and Alvorada do Oeste (2). Species with greatest distribution were C. foxi and C. leopoldoi. In Porto Velho were collected 1,773 individuals (1,179 female and 594 males), collected 228 individuals and 51 species in forest, and collected 1,545 individuals and 19 species in pasture. The most abundant species were C. insignis (n=1,472; 83%), C. foxi (n=45; 2.5%), C. glabrior (n=25; 1.4%) and C. ocumarensis (n=23; 1.3%). The forest presented most richness (41 species) and pasture presented greatest abundance (1,545 individuals). In Pasture the most abundant species were C. insignis (1,457-94.30%), C. foxi (34-2.20%) and C. ocumarensis (14-0.90%), and Forest the most abundant species were C. glabrior (25-10.96%), C. insignis (15-6.57%) e C. tetrathyris (15-6.57%). Diversity of species and eveness on Forest environment were greatest than Pasture environment. There was significant statistical difference (p=0.02) on species composition per environment. Diagnoses of some individuals of Rondônia were made. The findings of species of Culicoides on Rondônia now contribute to the total of 46 species, representing 36.8% of amazon species. The species C. darlingtonae, C. contubernalis e C. rodriguezi contribute for ampliation of geographic distribution this species and increase of amazon species fauna. The wide distribution of C. foxi and C. leopoldoi suggest that this species are ecletics for ecotypes and blood meal sources (chicken, livestock and human). The high abundance of C. insignis indicates that feeding behavior on mammals can be one of the factors contributing to the high number of individuals on light traps when placed near blood meal sources. Low number of individuals of C. paraensis on rural areas suggests that this specie is little attracted to light traps due to its diurnal or anthropophilic and urban habit that this species has. In Porto Velho, greatest abundance of C. insignis on Pasture can is related to the wet and muddy substrate with decaying organic matter, which favors the development of immature. The greatest diversity in forest can be explained by water bodies, wetlands, undergrowth, fruit peels present in this breeding sites, which makes it conducive to the proliferation and maintenance of Culicoides species. The diagnosis of the main morphotypes shows differences and similarities in relation to species already described, indicating possible new species for the state of Rondônia.
A família Ceratopogonidae possui quatro gêneros hematófagos: Forcipomyia, Leptoconops, Austroconops e Culicoides. Os Culicoides são insetos com até 3 mm de tamanho, as fêmeas são vetores de helmintos, protozoários e vírus. As principais arboviroses transmitidas são Oropouche, endêmica da bacia amazônica e Vírus da Língua Azul, que acomete bovinos silvestres e domésticos. As coletas de conveniência foram realizadas nos municípios de Alvorada do Oeste, Buritis, Cacoal, Costa Marques, Espigão do Oeste, Guajará-Mirim, Pimenta Bueno e São Francisco Guaporé, no estado de Rondônia, por meio de armadilhas luminosas HP entre 18:00 e 06:00. As coletas sistemáticas foram feitas no município de Porto Velho nos ambientes de floresta e pasto. Foram capturados 2.196 indivíduos, (1.515 fêmeas e 681 machos), distribuídos em 43 espécies e 27 morfótipos. As espécies mais frequentes foram Culicoides insignis (n=1.594; 72,59%), C. foxi (n=76; 3,46%), C. hildebrandoi (n=53; 2,41%), C. leopoldoi (n=44; 2%), C. ocumarensis (n=34; 1,55%), C. pusillus (n=29; 1,32%) e C. glabrior (n=28; 1,28%). Houve 37 novos registros de espécies em Rondônia e três novos registros para o Brasil. Os municípios com maior riqueza foram Porto Velho (56), Cacoal (26) e Costa Marques (14). Os municípios com menor de número de espécies foram Espigão do Oeste (4), Buritis (3) e Alvorada do Oeste (2). As espécies com maior distribuição foram C. foxi e C. leopoldoi. No município de Porto Velho foram capturados 1.773 indivíduos (1.179 fêmeas e 594 machos), sendo coletados 228 espécimens e 51 espécies em floresta, e 1.545 indivíduos e 19 espécies em pasto. As espécies mais abundantes foram C. insignis (1.472; 83%), C. foxi (45; 2,5%), C. glabrior (25; 1,4%) e C. ocumarensis (23; 1,3%). Floresta apresentou maior riqueza (41) e o pasto apresentou maior abundância (n=1.545). No pasto as espécies mais abundantes foram C. insignis (1.457-94,30%), C. foxi (34-2,20%) e C. ocumarensis (14-0,90%), e em floresta as mais abundantes foram C. glabrior (25-10,96%), C. insignis (15-6,57%) e C. tetrathyris (15-6,57%). A diversidade de espécies e a homogeneidade foram maiores em área de floresta do que o ambiente pasto. Houve diferença estatística significativa (p=0,02) na composição de espécies por ambiente. Foram realizadas diagnoses de alguns morfótipos de Rondônia. O achado de espécies de Culicoides em Rondônia contribui agora para o total de 46 espécies, representando 36,8% das espécies amazônicas. As espécies C. darlingtonae, C. contubernalis e C. rodriguezi contribuem para a ampliação da distribuição geográfica dessas espécies e o incremento da fauna de espécies amazônicas. A ampla distribuição de C. foxi e C. leopoldoi sugere que essas espécies são ecléticas às opções de ecótopos e fontes de repasto (galinhas, gados e humanos). A alta abundância de C. insignis indica que o comportamento alimentar sobre os mamíferos pode ser um dos fatores que contribui para o elevado número de indivíduos em armadilhas luminosas, quando colocadas próximas a fontes de repasto. O baixo número de C. paraensis na zona rural sugere que essa espécie é pouco atraída às armadilhas luminosas devido a seu hábito diurno ou ao hábito antropofílico e urbano que essa espécie apresenta. Em Porto Velho, a maior abundância de C. insignis em pasto pode está relacionado ao substrato úmido e enlameado com matéria orgânica em decomposição, que favorece o desenvolvimento de imaturos. A maior diversidade em floresta pode ser explicada pelos corpos d’água, brejos, serrapilheira, cascas de frutas presentes neste ecótopo, que torna propício à proliferação e manutenção de espécies de Culicoides. A diagnose dos principais morfótipos mostra diferenças e semelhanças em relação às espécies já descritas, indicando possíveis espécies novas para o estado de Rondônia.

Plusieurs systématiques se côtoient aujourd'hui : la systématique typologique, d'inspiration linnéenne, qui repose sur l'examen morphologique de types porte-nom. Depuis plusieurs années, une systématique phylogénétique conduit également à la création de taxons nouveaux, de niveau spécifique, infra-spécifique et supra-spécifique. Dans ce travail, nous avons cherché une approche globale, qualifiée de taxonomie intégrative couplant les approches morphologiques traditionnelles, la systématique moléculaire et la geomorphométrie alaire, dans le but d'éclairer la notion d'espèce chez les Culicoides avec ses corollaires épidémiologiques, étant donnée l'importance de ces insectes dans des maladies majeures d'intérêt vétérinaire telles que la fièvre catarrhale ovine (FCO) et la maladie causée par le virus de Schmallenberg (SBV). Cette approche nous a permis de mettre en lumière la complexité systématique des espèces affines (C.clastrieri/C.festivipennis, les groupes Obsoletus, Pulicaris et Vexans). De plus, nous nous sommes intéressés aux préférences trophiques des Culicoides de la région Champagne-Ardenne
Several systematic coexist today: typological systematic of Linnaean spirit, based on morphological examination of type specimens. For several years, a phylogenetic systematics also led to the creation of new taxa, at specific, sub-specific and supra-specific level.In this study, we have sought a comprehensive approach i.e. an integrative taxonomy that coupling traditional morphological approaches, molecular systematic and wing geometry morphometric in order to clarify the concept of species in the Culicoides with epidemiological corollaries, given the importance of these insects in major diseases of veterinary interest such as bluetongue (BT) and the disease caused by the Schmallenberg virus (SBV). This approach allowed us to highlight the systematic complexity of related species (C.clastrieri / C.festivipennis, the Obsoletus, Pulicaris and Vexans groups). In addition, we investigated the host preference of Culicoides of the Champagne-Ardenne region

Culicoides (Diptera: Ceratopogonidae) are small (<3mm) blood feeding flies. These flies are biological vectors of viruses, protozoa and filarial nematodes affecting birds, humans, and other animals. Among the viruses transmitted those causing bluetongue (BT), African horse sickness (AHS) and epizootic haemorrhagic disease (EHD) are of major veterinary significance. Culicoides (Avaritia) imicola Kieffer, a proven vector of both AHS and BT viruses, is the most abundant and wide spread livestock-associated Culicoides species in South Africa. Field isolations of virus and oral susceptibility studies, however, indicated that a second Avaritia species, C. bolitinos Meiswinkel may be a potential vector of both BT virus (BTV) and AHS virus (AHSV). Differences in oral susceptibility, which are under genetic control, of populations from different geographical areas to viruses may be an indication of genetic differences between these populations, which may be the result of limited contact between these populations. A good knowledge of the distribution, spread and genetic structure of the insect vector is essential in understanding AHS or BT disease epidemiology. In the present study, an effort was made to gather field specimens of both C. imicola and C. bolitinos from different areas within their natural distribution in South Africa. The aim was to partially sequence two mitochondrial genes from these specimens and to analyse the sequence data making use of phylogenetic trees to clarify the genetic relationships between individuals or groups collected from geographically distinct sites. The two species were collected from four geographically separated areas in South Africa viz. Gauteng Province, Eastern Cape Province, Western Cape Province as well as the Free State Province. DNA was extracted from a total of 120 individual midges of the two Culicoides species using DNA extraction kits. Extracted DNA was analysed using PCR, sequencing as well as phylogenetic methods. A total of 117 mitochondrial DNA COI and 104 mitochondrial 16S ribosomal RNA CulidoidesDissertation (MSc)--University of Pretoria, 2010.
Veterinary Tropical Diseases
unrestricted

Culicoides molestus (Skuse) is the major species of biting midge that plagues human comfort in the estuarine Gold Coast region of southeast Queensland. Local authorities have initiated a search for an effective, non-chemical means of control, that would minimize human-midge interaction. The effectiveness of a program to control an organism, such as a biting midge, is dependent upon knowledge of the biology of the particular organism of interest. This project revolved examines the lifecycle of C. molestus in detail. It addresses questions regarding the location, seasonal distribution, and dispersal of its juvenile stages in the sand of infested beaches, and their response to chemical treatment, the monthly and annual cycles of the adult midge, and the possibilities of achieving laboratory oviposition, as a first step to laboratory colonisation. The distribution of eggs, larvae and pupae of C. molestus was found to be mostly concentrated around, but below, mean tide level. They also occurred well below the mean tide level. Eggs and larvae have been recovered from as deep as 10 cm in the sand. A seasonal study of the juveniles of this species indicated that they were more strongly influenced by tides than seasons. After a routine pest-control larviciding treatment, a beach recolonisation study revealed that beaches become suitable for oviposition approximately two months after treatment. Large larvae invaded the sprayed areas within days of treatment, which suggests the existence of a refuge outside of the reach of the insecticide. Larvae found in clean (egg- and larva-free), isolated sand containers, placed on the study beach, indicated that larvae could swim in or on the water as a way of moving around the beach. Extended bite-rate studies highlighted the existence of four peaks in adult midge biting activity during the course of a year, around the mid seasons. The strongest peaks of activity were found to be in autumn and spring, but the data suggest that the species undergoes four generations in a year. Through a series of trial-and-error experiments, oviposition under laboratory conditions was achieved. Although the time from blood-feeding to egg maturation is not yet well determined, it occurs within an eight day mean survival period. Blood quality appears critical for adult blood-fed midge survival. Midges fed on the blood of a volunteer who was frequently exposed to midge bites do not live long enough to mature its eggs. The partial ovarial development of one unfed adult female, reared in the laboratory, indicates that C. molestus is facultatively anautogenous.

Culicoides molestus (Skuse) is the major species of biting midge that plagues human comfort in the estuarine Gold Coast region of southeast Queensland. Local authorities have initiated a search for an effective, non-chemical means of control, that would minimize human-midge interaction. The effectiveness of a program to control an organism, such as a biting midge, is dependent upon knowledge of the biology of the particular organism of interest. This project revolved examines the lifecycle of C. molestus in detail. It addresses questions regarding the location, seasonal distribution, and dispersal of its juvenile stages in the sand of infested beaches, and their response to chemical treatment, the monthly and annual cycles of the adult midge, and the possibilities of achieving laboratory oviposition, as a first step to laboratory colonisation. The distribution of eggs, larvae and pupae of C. molestus was found to be mostly concentrated around, but below, mean tide level. They also occurred well below the mean tide level. Eggs and larvae have been recovered from as deep as 10 cm in the sand. A seasonal study of the juveniles of this species indicated that they were more strongly influenced by tides than seasons. After a routine pest-control larviciding treatment, a beach recolonisation study revealed that beaches become suitable for oviposition approximately two months after treatment. Large larvae invaded the sprayed areas within days of treatment, which suggests the existence of a refuge outside of the reach of the insecticide. Larvae found in clean (egg- and larva-free), isolated sand containers, placed on the study beach, indicated that larvae could swim in or on the water as a way of moving around the beach. Extended bite-rate studies highlighted the existence of four peaks in adult midge biting activity during the course of a year, around the mid seasons. The strongest peaks of activity were found to be in autumn and spring, but the data suggest that the species undergoes four generations in a year. Through a series of trial-and-error experiments, oviposition under laboratory conditions was achieved. Although the time from blood-feeding to egg maturation is not yet well determined, it occurs within an eight day mean survival period. Blood quality appears critical for adult blood-fed midge survival. Midges fed on the blood of a volunteer who was frequently exposed to midge bites do not live long enough to mature its eggs. The partial ovarial development of one unfed adult female, reared in the laboratory, indicates that C. molestus is facultatively anautogenous.
Thesis (Masters)
Master of Philosophy (MPhil)
School of Environmental and Applied Science
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The biting midges Culicoides are vectors of some etiological agents to humans and other mammals. There are just a few studies in Amazon basin about that insects and the diversity and richness of this genus is underestimated. The aims of this study were to describe the fauna of midges in a rural settlement in Amazon and to understand how the anthropic effects may affect the richness and abundance of these putative local vectors. Collections were done by CDC light traps in some environment as primary forest, forest borders, secondary forest, fruit garden and peridomiciliary area with animal shelters. We identified 6.093 individuals, distributed in 42 species, 39 morphotypes of seven subgenera and /or species groups and five new species of Reticulatus group. We found 13 new occurrence to Amazonas State C. aldomani, C. batesi, C. brownie, C. debilipalpis, C. flavivenula , C. franklini, C. glabellus, C. guamai, C. guttatus, C. ocumarensi, C. paramaruim, C. pusilloides and C. tidwelli, and two new occourence to the country, the species C. brownie and C. tidwelli. Our results show that different anthropic levels affect the abundance and diversity of midges. The diversity was high in less anthropized environments. The peridomiciliary areas had a high richness and abundance of Culicoides.
Os Culicoides são insetos transmissores de agentes patogênicos ao homem e outros animais. Na região amazônica existem poucos estudos sobre esses insetos e o número de espécies é subestimado. Este estudo objetivou conhecer a fauna de Culicoides de um assentamento rural na Amazônia e verificar se o efeito antrópico afeta a diversidade e abundância dos mesmos. As coletas foram feitas com armadilhas luminosas do tipo CDC em ambientes de floresta, borda de floresta, capoeira e peridomicílio. Foram identificados 6.093 indivíduos, distribuídos em 42 espécies, 39 morfótipos de sete subgênero/grupo e cinco espécies novas do grupo Reticulatus. Foram encontrados 13 novos registros de espécies de Culicoides para o estado do Amazonas: C. aldomani, C. batesi, C. brownie, C. debilipalpis, C. flavivenula , C. franklini, C. glabellus, C. guamai, C. guttatus, C. ocumarensi, C. paramaruim, C. pusilloides e C. tidwelli, sendo C. brownie e C. tidwelli novos registros para o Brasil. Os resultados indicam que diferentes graus de antropização do ambiente influenciaram a abundância e diversidade dos Culicoides, cuja diversidade foi alta em ambientes menos antropizados. Ambientes totalmente antropizados, como peridomicílio, apresentaram maior abundância e riqueza.

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

The biting midges (family Ceratopogonidae) is one of the most species rich amongst the biting flies (Diptera) and has been recorded from most parts of the world. The species are mostly known for their capability to act as vectors for several important diseases, which have helped in shaping the focus to one of its genera, Culicoides Latreille, 1809.   This thesis gives an overview of the knowledge of the Swedish diversity, in the first paper (paper I) with a closer look at the species of Dasyhelea Kieffer, 1911 where all twenty species found in Sweden are presented with their associated localities, and two new species are described.  In the second paper (paper II) the biting midge diversity of Sweden is presented based on specimens collected from several localities. All these individuals were barcoded using the mitochondrial cytochrome oxidase I gene (COI). The analysis included 773 specimens that were assigned into 214 barcoding clusters (BINs) and sorted into 164 groups based on their morphology. The third paper (paper III) broadens the scale were the evolutionary relationships within the family are investigated by applying five protein coding genes (COI, CAD, TPI, AATS and PGD) and specimens from different parts of the World. The analysis recovers Ceratopogonini, Forcipomyia Meigen, 1818 and Bezzia Kieffer, 1899 as paraphyletic and Palpomyia Meigen, 1818 polyphyletic. In the last and fourth paper (paper IV) the family is used as a model organism together with Hymenoptera for an alternative analysis method for reducing the impact of saturation and long-branch attraction using non-synonymous coding (e.g. Degen1) on only parts of a dataset. The effectiveness of the method is compared to the removal of the faster evolving third codon position. The result yields a higher number of supported nodes as well as a higher median of support for the method as well as an ability to reduce long-branch attraction artifacts.

At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: Manuscript. Paper 3: Manuscript. Paper 4: Manuscript.

 

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A família Ceratopogonidae tem aproximadamente 5360 espécies, das quais 205 são extintas. Estas espécies estão distribuídas em 125 gêneros. As espécies de Culicoides estão entre as menores moscas hematófagas do mundo, medindo de um a três milimetros de comprimento. Mais de 1400 espécies têm sido identificadas em todo o mundo, das quais 96%, obrigatoriamente, sugam sangue de mamíferos (inclusive humanos) e aves, tendo sua principal importância como vetores de viroses humanas e de animais. Apesar da importância epidemiológica são poucos os trabalhos realizados no Brasil sobre biologia, comportamento de ataque e ecologia das espécies de Ceratopogonidae que sugam o homem. Este trabalho teve como objetivo fazer o levantamento das espécies de Ceratopogonidae que ocorrem em áreas costeiras do Estado do Pará, estudando as atividades horárias de ataque das espécies a procura do repasto sanguíneo no ser humano e fornecer informações básicas importantes para definição de períodos menos e mais problemáticos por causa do ataque destes dípteros. Foram dois os pontos amostrados na região costeira do Estado do Pará, um no litoral atlântico a nordeste do estado do Pará (município de Marapanim) e outro na área costeira do estuário do rio Pará (Outeiro, município de Belém). As coletas foram realizadas mensalmente de abril e outubro de 2003, utilizando-se isca humana e armadilhas CDC. Foram coletados 4083 exemplares de Ceratopogonidae distribuídos em 29 espécies pertencentes a seis gêneros, sendo 620 exemplares provenientes de Outeiro distribuídos em 21 espécies, pertencentes a cinco gêneros e 3463 exemplares provenientes de Marapanim de nove espécies, pertencentes a quatro gêneros. A fauna da região litorânea mostrou-se bastante diferente da zona do estuário do rio Pará, sendo Leptoconops brasiliensis a única espécie comum aos dois pontos amostrados. Em Outeiro as espécies mais abundantes foram Culicoides batesi e Culicoides denisae e em Marapanim Culicoides maruim e Culicoides phiebotomus. As espécies Culicoides crucifèr e Culicoides daviesi coletadas em Outeiro são registradas pela primeira vez no Brasil e Culicoides denisae e Culicoides phlebotomus tiveram suas distribuições ampliadas ao estado do Pará. Em Outeiro encontrou-se correlação negativa significativa entre a temperatura e a atividade de antropofilia por horas do dia de Culicoides batesi, Culicoides cruciftr e Culicoides paramaruim e correlação positiva significativa com a umidade relativa do ar. Em Marapanim Culicoides maruim apresentou correlação negativa significativa com a temperatura e positiva significativa com a umidade do ar. Em Outeiro, de acordo com os horários de ataque das duas espécies mais abundantes (C. batesi e C. denisae), principalmente, durante o período seco do ano, as pessoas são mais incomodadas pelos maruins pela parte da manhã das 5:00 as 9:00h e menos intensamente no período da tarde a partir das 16:00h e se estendendo pela noite. Em Marapanim durante o período seco os maruins não causam problemas aos visitantes, pois a espécie mais abundante (C. maruim) ocorreu preferencialmente nos primeiros horários do dia e é restrita a áreas de mangue, porém esta causa ataques vorazes as pessoas que precisam entrar nos manguezais. Entretanto, os visitantes podem ter problemas no período chuvoso com a espécie C. phlebotomus, que ocorre também em áreas de praia; durante este estudo ocorreu em todos os meses de coleta em baixos números de exemplares, próximo ao mangue, sendo mais abundante em junho (início do período seco) e apresentou correlação positiva significativa com a umidade do ar. A coleção de Invertebrados do Museu Paraense Emílio Goeldi foi enriquecida com o material deste trabalho, acondicionado em lâminas e em pequenos tubos de ensaio.
The dipterous family Ceratopogonidae has 125 genera and approximately 5360 species, 205 of which are extinct. More than 1400 species of Culicoides are known worldwide, and these are among the smallest blood-sucking flies in the world, measuring one to three mm in length. Some 96% of these species are obligate blood feeders on mammals (including human) and birds, and they are important vectors of human and animal viroses. Inspite of their epidemiological importance, few studies have been made of the biting behaviour and general ecology of Brazilian ceratopogonid specíes. This study surveyed the ceratopogonid species in two coastal areas of Pará State in northern Brazil, with reference to the hotus of the day when humans were bitten. The two coastal sampled were: the city of Marapanim on the northeastern Atlantic coast of Pará and Outeiro, a district of Belém on the Pará River estuary. Collections were made monthly between April and October 2003, using human bait and CDC traps. In total, 4083 specimes of Ceratopogonidae of 29 species in six genera were collected, 620 (21 species in five genera) from Outeiro and 3463 specimes (9 species in 4 genera) from Maripanim. The two faunae were seen to be sufficiently different from one another Leptoconops brasiliensis was the only species common to both sites. In Outeiro, the most abundant species were Culicoides batesi and C. denisae while in Marapanim these were C. maruim and C. phkbotomus. C. crucifèr and C. daviesi were collected in Outeiro and are registered for the first time fmm Brazil. C. denisae, C. phkbotomus and L. brasiliensis are reported for the first time for the state of Pará. In Outeiro, a significant negative correlation was found between air temperature and biting activity of C. batesi, C. cructfer and C. paramaruim. A significam positive correlation was found for biting activity and air relative humidity. In Marapanim, C. maruim biting activity showed a significant negative correlation with air temperature and significant positive with the air relative. The collected material, mounted on slides or in specimen tubes, is deposited in the Invertebrate Collection of the Museum Paranese Emílio Goeldi, Belém, Brazil.

La Fièvre catarrhale ovine (FCO) est une arbovirose émergente en Europe depuis la fin desannées 90. Elle affecte principalement les ruminants par la piqûre de petits moucheronshématophages, les Culicoides (Diptera : Ceratopogonidae). Pendant l’été 2006, l’introductiondu sérotype 8 de la FCO, dans la région de Maastricht (Pays-Bas) a rapidement diffusé dansles Ardennes, générant de lourdes pertes pour les éleveurs de bovins et d’ovins. Cesévènements interrogent sur la capacité des Culicoides de la région paléarctique à transmettrela FCO. Ils révèlent la nécessité de mieux connaître la biologie de ces diptères.Nous avons développé successivement dans ce travail, trois axes de recherche qui se sontappuyés sur un travail de terrain réalisé principalement au sein de deux élevages situés dansles Ardennes françaises.Dans un premier temps, nous avons réalisé une expérimentation de gorgement de Culicoidesde captures et d’émergences, provenant des Ardennes, sur petits ruminants virémiques pour leBTV8. A l’issue des expérimentations, une femelle gorgée de l’espèce Culicoides obsoletus apondu et a été retrouvée faiblement positive lors de la recherche du génome du virus de laFCO. Les résultats obtenus ainsi que les difficultés rencontrées lors de la réalisation de cetype d’expérimentation sont discutés.Le deuxième travail exposé s’est intéressé au comportement trophique des Culicoides parl’étude de l’origine du repas sanguin de femelles de Culicoides piégées dans des biotopesvariés. A cette fin, nous avons utilisé des marqueurs moléculaires pour amplifier l’ADN devertébré présent dans les estomacs de femelles gorgées. Ces analyses ont permis de mettre enévidence que des espèces appartenant aux complexes Obsoletus, Pulicaris, ou encore,Culicoides dewulfi, avaient un spectre d’hôte large. Certaines d’entre elles peuvent se gorger àla fois sur les ruminants domestiques et sur la faune sauvage. De plus, ce type d’étuderenseigne sur l’écologie des différentes espèces de Culicoides.Enfin, nous présentons les résultats d’une étude faunistique fondée sur des captures avec despièges lumineux, mais aussi, des prélèvements de boue pour la recherche des gîtes larvaires.Les résultats de piégeages entre les deux exploitations ont été comparés, notamment en termesde biodiversité, et sont discutés en regard des différences de pratiques d’élevage entre lesdeux exploitations choisies d’une part, et la mise en évidence des gîtes larvaires d’autre part.De nombreuses espèces de Culicoides ont émergé au laboratoire à partir des prélèvements deboues, qui ont été caractérisés macroscopiquement. Les gîtes larvaires de C. obsoletus, peuconnus jusqu’alors, ont été mis en évidence dans les deux fermes. Ils ont fait l’objet d’un suivisur plusieurs mois.L’ensemble de ces études contribue à la meilleure connaissance des Culicoides présents dansles Ardennes et de leur biologie, elles permettent de rendre compte des espèces qui semblenttrès inféodées à l’élevage de bovins, et celles qui sont plus ubiquistes. Certains travauxprésentés pourraient être poursuivis pour mettre en évidence les espèces ou populations deCulicoides plutôt sylvatiques, et pour mettre en place de nouvelles expérimentations sur lacompétence et la capacité vectorielle des Culicoides
Since the late 90’s, Bluetongue disease (BT) can be considered as an emerging arbovirose inEurope. This disease is mainly transmitted to ruminants by the bites of minute size midges,the Culicoides (Diptera: Ceratopogonidae), also known as biting midges. An outbreak of BTserotype 8 occurred during summer 2006, in the region of Maastricht (Netherlands) andspread quickly to the Ardennes region. The epizooty lead to severe losses in cattle and sheepholdings. These events highlighted the lack of knowledge on the vectorial capacity ofpaleartic Culicoides species, and more generally on their biology.Three approaches are successively treated in this document. They are all based on field workconducted mainly in two holdings located in the Ardennes region.First, an experiment to assess oral susceptibility of Culicoides to Bluetongue virus (BTV) 8was undertaken. Field collected and emerging Culicoides coming from the Ardennes wereengorged on viremic small ruminants. At the end of the experiments, one Culicoides obsoletusfemale was found bloodfed and laid eggs. She was tested for BTV and was found weaklypositive for BTV genome. This result and the difficulties met during the experiment havebeen discussed.The second study focused on the bloodmeal origin of engorged females of Culicoides. Thesewere collected by light traps set in different kinds of environment. Molecular markers wereused in order to amplify the DNA of vertebrates present in the stomach of bloodfed females.Some of the species processed belonging to the Obsoletus or the Pulicaris complex, andCulicoides dewulfi fed on a wide variety of hosts, including domestic ruminants and wildanimals. Moreover, this kind of study brings information on the ecology of different speciesof Culicoides.Finally, a faunistic survey is presented. It was achieved through light trap collections ofmidges and also thanks to the sampling of potential breeding sites. Biodiversity in thecollection of midges captured by light traps between the two holdings were compared.Differences observed are discussed taking into account the differences in breeding practicesbetween the two holdings and the breeding sites investigations. Numerous species ofCulicoides emerged in the laboratory from soil samples which were macroscopicallydescribed. Breeding sites of C. obsoletus, which were not well documentated in the literature,were found in both farms. These were monitored over some months.This work contributes to a better knowledge of the Culicoides present in the Ardennes andtheir biology. It highlights the species which are closely related to the cattle holdingenvironment, and those which are ubiquist. Some of these studies could be continued in orderto highlight the species more related to the forested areas, and to set new experiments onvectorial competence and capacity

Tese de Doutoramento em Ciências Veterinárias, especialidade de Sanidade Animal
Culicoides genus is of major importance in animal and human health since hematophagous females are vectors of several pathogens, like viruses (Bluetongue, African Horse Sickness and Schmallenberg) and filarial nematodes, among others. As a consequence, female biting midges are responsible for huge economical losses worldwide. A deeper knowledge of Culicoides fauna present in each country and their ecological preferences is required, so different control strategies can be applied efficiently. The present work was based on Culicoides species captures during the National Entomologic Surveillance Program for Bluetongue disease in mainland Portugal and Azores and Madeira archipelagos (2005-2013), using miniature CDC light traps, and in 2015, with OVI traps, in the framework of VectorNet European network. Biting midges were evaluated phenotypically and genetically, showing the variation of Culicoides species in Portuguese territory. Twenty-two Culicoides species were mentioned for the first time in Portugal, including C. dewulfi and C. montanus in mainland Portugal and species of Obsoletus group in the islands of Azores archipelago where they were never reported, as well as a description of Culicoides paradoxalis, a new species for science. Moreover, a detailed study focused the morphology, genetics and ecology of Obsoletus group, showing that the distribution of those species was unequal in mainland Portugal. Plus, a redefinition of the 3rd palpus segment length/width ratio and spermathecae size intervals, aiming Obsoletus group species identification, together with the reference of anatomical aberrations in these species, was performed, allowing the reduction of errors during midges studies. An identification key for all known Portuguese midges was also created. In this thesis, elaboration of risk assessment maps based on the association of some abiotic variables with the occurrence of C. imicola, C. pulicaris, C. punctatus, C. newsteadi and species from Obsoletus group in mainland Portugal were also performed. Finally, evaluation of C. imicola morphological modifications in sensorial organs localized in the 3rd palpus segment, used for host detection, was performed after an assay with pyrethroid insecticides (permethrin and deltamethrin) at different concentrations, showing complete destruction of sensorial organs with probable feeding implications. The results presented in this scientific work contributed to a better knowledge of Culicoides genus in Portugal, being some of them of worldwide relevance.
RESUMO - Caracterização fenotípica e genética de Culicoides (DIPTERA: CERATOPOGONIDAE) em Portugal e comparação do efeito de inseticidas piretróides no seu controlo - Os insetos do género Culicoides (Diptera: Ceratopogonidae) possuem uma grande importância em Saúde Animal e Humana, uma vez que as fêmeas hematófagas são vetores de vários agentes patogénicos, como vírus (da Língua Azul, da Peste Equina Africana, da Doença Hemorrágica Epizoótica e de Schmallenberg), filarídeos, entre outros. Consequentemente, as fêmeas do género Culicoides são responsáveis por elevadas perdas económicas a nível mundial. Um conhecimento profundo da fauna de Culicoides presente em cada país é necessário, assim como as suas preferências ecológicas, de modo a que possam ser aplicadas de modo mais eficaz diferentes estratégias de controlo. Uma das doenças que nos últimos anos afetou gravemente os ovinos em Portugal foi a Doença da Língua Azul. O primeiro foco desta doença em Portugal foi reportado em julho de 1956, afetando os animais da região sul do território abaixo do rio Tejo, excluindo a região algarvia, tendo sido causado pelo serótipo 10 (BTV-10). O país foi considerado livre da doença em 1960 depois da aplicação de uma vacina em ovinos. Nesse período de 4 anos, a doença matou mais de 179 mil ovinos. A Peste Equina Africana foi detetada em Portugal em Agosto de 1989 em Castro Marim, tendo-se alastrado pelo Alentejo, ao longo da zona fronteiriça. As medidas de controlo incluiram o estudo entomológico de insetos do género Culicoides, vacinação massiva dos cavalos, burros e mulas, aspersão de inseticidas nas cavalariças e estábulos e divulgação de informação aos proprietários dos animais sobre prevenção. Até ao final de 1989 a doença vitimou 202 cavalos na zona do rio Guadiana, tendo sido Portugal considerado livre da doença em 1992. Após um silêncio epizoótico de 44 anos, a doença da Língua Azul voltou a surgir em território nacional, nas regiões centro-oeste e sul, em novembro de 2004, sendo desta vez causada pelo serótipo BTV-4. EM 2005, foi criado o Programa Entomológico de Vigilância Nacional para a doença da Língua Azul em Portugal Continental e nos arquipélagos dos Açores e da Madeira (DGV/FMV, 2005-2013) com o objetivo de conhecer a distribuição de Culicoides em território português, reportando às autoridades competentes a presença de espécies com capacidade vetorial, por forma a que se pudesse atuar em tempo real e prevenir a dispersão da Língua Azul em caso de um surto da mesma. Em setembro de 2007, o serótipo BTV-1 foi detetado num concelho do Alentejo perto da fronteira com Espanha. Entre setembro e dezembro de 2007, o serótipo 1 (BTV-1) foi assinalado nas regiões do centro e sul de Portugal continental, tendo-se expandido para o norte do território desde 2008 e mostrando uma diferente distribuição quando comparada com a dos serótipos BTV-4 e BTV-10. As campanhas de vacinação contra os serótipos 4 (2005-2008) e 1 (2007-2010) do vírus da doença da Língua Azul, em ovinos e bovinos, contribuíram para o controlo da emergência e dispersão da doença. Em 2013, o serótipo BTV-4 reemergiu no nosso país, tendo sido adotadas medidas de controlo da doença, que passaram pela vacinação obrigatória dos ovinos e facultativa dos bovinos na região do Algarve contra o referido serótipo. A vacinação contra o mesmo serótipo foi permitida na região do Alentejo como medida profilática. Finalmente, em setembro de 2015, e após um silêncio epizoótico de 3 anos, o serótipo 1 foi novamente detetado na região do Alentejo, nos concelhos de Serpa, Moura e Barrancos. Presentemente, o serótipo 1 encontra-se presente em todo o território português enquanto que o BTV-4 encontra-se em circulação apenas na região do Algarve. Durante o Programa Entomológico de Vigilância Nacional para a doença da Língua Azul em Portugal Continental e nos arquipélagos dos Açores e da Madeira (2005-2013), com recurso a armadilhas do tipo CDC, e em 2015 com armadilhas OVI, no âmbito do grupo de trabalho europeu VectorNet, foram realizadas capturas de insetos do género Culicoides que permitiram a realização do presente trabalho. Os insetos capturados foram analisados fenotípica e geneticamente, mostrando a variação das espécies de Culicoides em território português. C. imicola foi a espécie mais capturada, representando 70,92% do total estimado de insetos capturados. Seguidamente, surgiu C. achrayi (10,34%), C. punctatus (9,34%), espécies do grupo Obsoletus (4,93%) e C. newsteadi (1,93%) como as espécies mais capturadas. C. pulicaris, espécie vetor do vírus da Língua Azul, perfez 0,08% do total estimado de insetos capturados (N=4.384.502). A distribuição espacial das espécies de Culicoides em Portugal Continental apresentou padrões diferentes. Assim, enquanto que algumas espécies deste género estavam dispersas por todo o território (e.g., C. achrayi, C. punctatus), outras localizaram-se preferencialmente nas regiões norte e centro (e.g., C. deltus, C. heliophilus), diversas nas regiões centro e sul (e.g., C. nubeculosus, C. sahariensis) e algumas foram capturadas consistentemente em regiões específicas (e.g., C. impunctatus e C. lupicaris). Vinte e duas espécies de Culicoides foram mencionadas pela primeira vez em Portugal: C. alazanicus, C. deltus, C. dewulfi, C. heliophilus, C. jumineri near C. bahrainensis, C. jurensis, C. kingi, C. lupicaris, C. malevillei, C. montanus, C. paolae, C. picturatus, C. remmi, C. riebi, C. santonicus, C. semimaculatus, C. simulator e C. subfagineus em Portugal continental, C. obsoletus, C. scoticus nas ilhas do arquipélago dos Açores onde ainda não tinham sido referidas e C. circumscriptus e C. newsteadi em todas as ilhas do mesmo arquipélago, com exceção das Flores e do Corvo. Para além destas, foi realizada a descrição de uma nova espécie para a ciência, Culicoides paradoxalis. Outro objetivo deste trabalho focou a morfologia, a genética e a ecologia do grupo Obsoletus, mostrando que a distribuição destas espécies é diferente em Portugal continental. Entre as quatro espécies presentes, C. dewulfi foi apenas identificada em três explorações no Norte do território português. As restantes três espécies do grupo Obsoletus encontravam-se presentes em todo o país, sendo C. obsoletus a espécie mais prevalente no Norte, Centro Sul e Sul de Portugal continental, enquanto que C. scoticus foi a espécie mais prevalente na região Centro Norte. Esta espécie foi menos comum que C. obsoletus e C. montanus nas regiões Centro Sul e Sul. Estas três espécies estão bem adaptadas ao território de Portugal Continental, com a exceção da região Centro Sul. A correta identificação de espécies do complexo Obsoletus nem sempre é possível devido às suas elevadas semelhanças morfológicas. Nesse sentido, foi realizado um estudo com base em várias estruturas anatómicas destes insetos, que levaram à redefinição dos intervalos relativos ao rácio comprimento/largura do 3.º segmento do palpo e ao comprimento das espermatecas. Para além disso, a menção de várias aberrações anatómicas nestas espécies (fossetas sensoriais aberrantes, fossetas sensoriais duplas, segmentos do palpo fundidos, segmentos do palpo com alterações morfológicas, número irregular de espermatecas, espécimens com genitália masculina e simultaneamente com espermatecas, comprimento do 3.º artículo do palpo desigual dentro do mesmo exemplar, flagelómeros antenares fundidos e aberrantes) permitirá uma redução nos erros associados ao estudo destes insetos. Uma chave de identificação para todas as espécies de Culicoides presentes em Portugal foi também elaborada durante este estudo que incidiu sobre cerca de 93.100 exemplares deste género. Nesta tese, a produção de mapas de análise de risco baseados na associação de algumas variáveis abióticas climáticas (temperatura média no período mais seco e no período mais húmido) e edáficas (territórios artificializados, áreas agrícolas e agroflorestais, florestas e meios naturais e seminaturais, zonas húmidas e corpos de água) com a ocorrência de C. imicola, C. pulicaris, C. newsteadi, C. punctatus e espécies do grupo Obsoletus em Portugal Continental foi também efetuada. A partir destes mapas observou-se que a probabilidade de capturar C. imicola é maior nas regiões a sul do rio Tejo e na Beira Baixa, enquanto que C. pulicaris ocorre com maior frequência nas regiões do norte de Portugal continental, na linha costeira e em zonas com elevada altitude. A probabilidade de ocorrência de C. newsteadi é semelhante à de C. imicola, embora aquela espécie possa ser capturada em regiões ligeiramente mais a norte que esta última. C. punctatus pode ocorrer com uma probabilidade maior que 50% em qualquer parte de Portugal Continental e em quase todas as estações do ano. As espécies do grupo Obsoletus surgem nas regiões do norte de Portugal continental, estando estas espécies praticamente ausentes na região Centro Sul durante o outono e inverno. A distribuição do grupo Obsoletus é aproximadamente a oposta de C. imicola. Finalmente, a avaliação de modificações morfológicas dos órgãos sensoriais localizados no 3.º segmento do palpo de C. imicola, usados na deteção do hospedeiro, foi realizada através de microscopia eletrónica de varrimento após um ensaio com inseticidas piretróides (permetrina e deltametrina) em diferentes concentrações. Este estudo revelou a completa destruição dos órgãos sensoriais, com uma provável influência na alimentação destes insetos. Tendo em conta o impacto económico associado aos agentes patogénicos que transmitem, aliado à descoberta recente de novos serótipos do vírus da Língua Azul na Europa (BTV-25 e BTV-27), os resultados apresentados neste trabalho científico evidenciam e sustentam a importância do estudo entomológico dos insetos do género Culicoides.
This work was partially funded by the Centre de Coopération Internationale en Recherche Agronomique pour le Développement (CIRAD), Ministère en charge de l’Agriculture (France) and by the convention Cherchuer d’Avenir 2011 from Languedoc-Roussillon region (France).

Les invasions biologiques constituent une source de préoccupation majeure du fait des conséquences écologiques, économiques et sanitaires dont elles sont responsables. Déterminer et comprendre les facteurs sous-jacents au succès invasif des espèces envahissantes permet de prédire de nouvelles invasions et de mettre en place des stratégies de contrôle. Culicoides imicola est un vecteur majeur d’Orbivirus d’intérêt vétérinaire incluant le virus de la fièvre catarrhale ovine (FCO). Suite à l’émergence de la FCO dans le bassin méditerranéen, les populations de C. imicola ont été découvertes dans des territoires où elles étaient considérées comme absentes, caractérisant alors cette présence comme la résultante d’une expansion récente de l’espèce. Cette thèse décrit un ensemble de travaux visant à comprendre l’histoire de la colonisation du bassin méditerranéen par C. imicola. L’utilisation d’une approche multi-marqueurs combinant des analyses de génétique de populations, des inférences basées sur la méthode Approximate Bayesian Computation (ABC) et la simulation mathématique de la dispersion atmosphérique de l’espèce, a permis (i) de déterminer l’origine des populations installées au Maghreb et au Moyen Orient et de décrire les routes de colonisation et la chronologie de ces évènements, (ii) de définir les caractéristiques démographiques, évolutives et temporelles de la colonisation du sud de l’Europe et (iii) de caractériser les principaux facteurs expliquant le succès d’expansion géographique des populations installées. Les principaux résultats de cette thèse permettent de proposer des hypothèses pour expliquer le succès de l’installation des populations de C. imicola dans le bassin méditerranéen
Biological invasions are of major concern because of their environmental, economic and health consequences. Determining and understanding the factors underlying the invasion success of species allow predicting potential other biological invasions, and developing vector control strategies. Culicoides imicola is a major vector species of Orbivirus, including the bluetongue virus (BTV) which affects domestic ruminants. Following BT emergence in the Mediterranean basin, C. imicola populations were recorded in territories where the species was considered to be absent, and consequently was described as expanding its range expansion on a short period. This Phd work describes a set of studies aiming at understanding the colonization history of the Mediterranean basin by C. imicola. The use of a multi-loci approach combining population genetics analyses, Approximate Bayesian Computation (ABC) methods and mathematical simulations of the atmospheric dispersion of the species enabled to (i) determine the origin of the established populations in the Maghreb and the Middle-East and describe the routes of colonization and the chronology of such events, (ii) define the demographic, evolutionary and temporal characteristics of south-western Europe colonization and (iii) characterize the main factors explaining the successful range expansion of the established populations. The main results of this thesis allow suggesting hypotheses to explain the successful establishment of C. imicola populations in the Mediterranean basin

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.

[spa] . Introducción: Las hembras de varias especies de insectos del género Culicoides (Diptera; Ceratopogonidae) transmiten arbovirus que afectan a rumiantes domésticos y salvajes tales como el virus de la Lengua Azul, de la Peste Equina Africana, el virus de Schmallenberg y el virus de la enfermedad hemorrágica epizoótica. La llegada y expansión de enfermedades de origen tropical o subtropical transmitidas por insectos vectores causan importantes problemas tanto médico-veterinarios como económicos, por lo tanto, el conocimiento de la biología y ecología de esos vectores es crucial para comprender la transmisión de arbovirosis y otros patógenos. Gracias a esos conocimientos podemos actuar en el momento preciso poniendo en marcha las medidas necesarias para su control y así evitar una posible epidemia o minimizar sus efectos. . Contenido de la investigación: El presente trabajo tiene dos objetivos fundamentales: 1.Estudiar la dinámica poblacional y fenología de especies de Culicoides vectores del virus de la Lengua Azul en España, donde se analizará la distribución y estacionalidad de hembras paras y nulíparas; se determinarán los factores ambientales implicados en la fenología de estas especies gracias a Modelos Bayesianos Generalizados Lineales Mixtos (GLMM) y se estudiaran los factores ambientales incluyendo el efecto indirecto de la abundancia poblacional de Culicoides sobre el periodo de actividad de hembras nulíparas y paras gracias a los Análisis de Trayectorias (Path analysis models). 2.Estudiar la bionomía de Culicoides asociados a granjas en las Islas Baleares, donde se determinarán los valores bionómicos básicos de diferentes especies en condiciones de laboratorio comparando distintos métodos con la finalidad de establecer una futura colonia en cautividad; además, se compararán resultados bionómicos de individuos a distintas temperaturas utilizando dos poblaciones diferentes. . Conclusión El análisis de la variación estacional latitudinal de la PF demostró que C. imicola no está presente en las provincias del norte mientras que las especies del complejo Obsoletus son las mayoritarias en dichas provincias. Además, se han encontrado provincias en las que hubo periodos del año donde no se capturaron individuos de ninguna especie vector, lo cual se debe tener en cuenta a la hora de calcular el Periodo Estacionalmente Libre de Vectores. C. newsteadi y C. pulicaris mostraron la población más elevada en Toledo, posiblemente debido a su preferencia a las zonas de interior. Culicoides imicola mostró que su periodo estacional fue más largo en zonas poco elevadas y más corto en zonas con una elevada acumulación de días con temperaturas sobre los 10ºC y precipitaciones altas durante el invierno. Para las especies del complejo Obsoletus, el periodo de actividad de las hembras adultas también fue más prolongado en zonas poco elevadas con mayor número de horas de sol y temperaturas más cálidas en primavera y otoño, así como en zonas con altas precipitaciones en otoño y gran abundancia de ganado bovino. Las hembras de C. newsteadi se relacionaron con inviernos cálidos, otoños cálidos con altas precipitaciones y áreas agroforestales con vegetación esclerófila y poca pendiente en el terreno. C. pulicaris mostró períodos de adultos más largos en sitios con un elevado número de días con temperaturas mayores a 10ºC durante el invierno. Las especies C. paolae y C. circumscriptus parecieron ser las más adecuadas para la cría en condiciones de laboratorio debido a sus tasas elevadas de oviposición, ciclo de vida corto, elevada supervivencia de la fase adulta y elevado porcentaje de hembras en la progenie. Temperaturas bajas mostraron una mayor tasa de supervivencia de adultos, un tiempo de oviposición más largo y periodos más largos de las fases inmaduras; mientras que las temperaturas elevadas aumentaron el número de huevos, el porcentaje de pupado, la emergencia de adultos y la velocidad de crecimiento de las larvas.
[cat] Introducció: Les femelles de diverses espècies d'insectes del gènere Culicoides (Diptera; Ceratopogonidae) transmeten arbovirus que afecten a remugants domèstics i salvatges com ara el virus de la Llengua Blava, el de la Pesta Equina Africana, el virus de Schmallenberg i el virus de la malaltia hemorràgica epizoòtica. L'arribada i expansió de malalties d'origen tropical o subtropical transmeses per insectes vectors causen importants problemes tant mèdic-veterinaris com econòmics, per tant, el coneixement de la biologia i ecologia d'aquests vectors és crucial per comprendre la transmissió d’arbovirosis i altres patògens. Gràcies a aquests coneixements podem actuar en el moment precís posant en marxa les mesures necessàries per al seu control i així evitar una possible epidèmia o minimitzar-ne els seus efectes. . Contingut de la investigació: El present treball té dos objectius fonamentals: 1. Estudiar la dinàmica poblacional i fenologia d'espècies de Culicoides vectors del virus de la Llengua Blava a Espanya, on s'analitzarà la distribució i estacionalitat de femelles pares i nul·lípares; es determinaran els factors ambientals implicats en la fenologia d'aquestes espècies gràcies a Models Bayesians Generalitzats Lineals Mixtes (GLMM) i s'estudiaran els factors ambientals incloent l'efecte indirecte de l'abundància poblacional de Culicoides sobre el període d'activitat de femelles nul·lípares i pares gràcies al Anàlisis de Trajectòries (Path analysis models). 2. Estudiar la bionomia de Culicoides associats a granges a les Illes Balears, on es determinaran els valors bionòmics bàsics de diferents espècies en condicions de laboratori comparant diferents mètodes, amb la finalitat d'establir una futura colònia en captivitat; a més, es compararan resultats bionòmics d'individus a diferents temperatures emprant dues poblacions diferents. . Conclusió: L'anàlisi de la variació estacional latitudinal de la PF va demostrar que a les províncies del nord d’Espanya C. imicola no esta present mentre que les espècies del complex Obsoletus son les espècies majoritàries a aquestes províncies. A més, s’han trobat províncies on hi va haver períodes de l'any on no es va capturar cap individu de les espècies vectors de Llengua Blava, la qual cosa s'ha de tenir en compte a l'hora de calcular el Període Estacionalment Lliure de Vectors. C. newsteadi i C. pulicaris foren igualment presents a totes les províncies analitzades mostrant la població més elevada a Toledo, possiblement a causa de la preferència d’aquestes espècies per a les zones d'interior. Culicoides imicola va mostrar que el seu període estacional va ser més llarg en zones poc elevades y més curt a zones amb major acumulació de dies amb temperatures sobre els 10ºC i precipitacions elevades durant l'hivern. Per a les espècies del complex Obsoletus, el període d'activitat de les femelles adultes també va ser més perllongat en zones poc elevades amb major nombre d’hores de sol i temperatures mitjanes més càlides a la primavera i a la tardor; així com en zones amb altes precipitacions a la tardor i gran abundància de bestiar boví. Les femelles de C. newsteadi foren relacionats amb hiverns càlids, tardors càlides amb altes precipitacions i àrees agroforestals amb vegetació escleròfil·la i poca pendent en el terreny. D'altra banda, C. pulicaris va mostrar períodes més llargs dels adults en llocs amb alta acumulació de dies sobre els 10ºC durant l'hivern. Les espècies C. paolae i C. circumscriptus van semblar ser les més adequades per a la cria en condicions de laboratori degut a les seves elevades taxes d‘oviposició, cicle de vida curt, elevada supervivència de la fase adulta i elevat percentatge de femelles en la progènie. Les temperatures baixes van augmentar la taxa supervivència dels adults i el temps de oviposició, a més d’un augment dels períodes de les fases juvenils; mentre que temperatures elevades van augmentar el nombre d'ous, el percentatge de pupat, l'emergència d'adults i la velocitat del creixement larvari.
[eng] . Introduction: Females of several biting midges species from genus Culicoides (Diptera; Ceratopogonidae) transmit arboviruses affecting wild and domestic ruminants such as Bluetongue, African Horse sickness, Schmallenberg virus and Epizootic Haemorrhagic disease viruses. The arrival and expansion of tropical or subtropical diseases transmitted by insect vectors cause important medical-veterinary and economic impacts; therefore, the knowledge of the biology and ecology of these vectors is crucial to understand the transmission of arbovirosis and other pathogens. Then, we can implement the necessary measures at the correct timing for its control, avoiding possible outbreaks or minimize its effects. . Research content: This work has two main objectives: 1. A study of the population dynamics and phenology of Culicoides Bluetongue virus vector species in Spain; an analysis of the distribution and seasonality of parous and nulliparous females; the environmental factors involved in the phenology of these species determined with Mixed Linear Generalized Bayesian Models (GLMM) and a study of environmental factors including the indirect effect of Culicoides population abundance on the period of activity of parous and nulliparous females through Path Analysis models. 2. A study of the bionomics of Culicoides associated with farms in the Balearic Islands. The basic bionomic values of different species in laboratory conditions were determined by comparing different methods to establish a future colony in the laboratory; in addition, we compared bionomic results at different temperatures using two different populations . Conclusion The analysis of the latitudinal seasonal variation of PF in Spain has demonstrated that Northern provinces showed absence of C. imicola during the whole year while the Obsoletus species were more present in Northern areas. In fact, depending on the province and the species, there were periods of the year were no individuals of any vector species were collected, which should be taken into account in order to calculate the Seasonally Vector-Free Period. C. newsteadi and C. pulicaris were equally present in all analysed provinces showing the highest population in Toledo, probably related to their inland preferences. For C. imicola, the length of the female adult season was longest in sites at low elevation and shortest with more accumulated degree days over 10ºC and higher rainfall during winter. For Obsoletus complex species, female adult seasons were also longest in sites at low elevation, with long spring and autumn daylight hours and warmer spring and autumn average temperatures as well as in sites with high precipitation in autumn and high abundance of cattle. Culicoides newsteadi showed early appearance and long adult seasons in sites with warm winters and autumns and high precipitation in autumn, and in sites with higher coverage of agro-forestry with schlerophyllous vegetation and gently sloping land. Meanwhile, C. pulicaris showed longer adult periods in sites with high number of accumulated degree days over 10ºC during winter. Culicoides paolae and C. circumscriptus seemed to be the most suitable for laboratory rearing due to their high oviposition rates, short life-cycle, long adult lifespan and female biased sex ratio. Low temperatures increased the adult lifespan, time to oviposit and life-cycle development whereas high temperatures increased the number of eggs, percentages of pupation and adult emergence as well as the larvae growth rate.

Les zoonoses constituent plus des deux tiers des pathologies virales qui concernent l’homme. Le développement et la démocratisation des outils de métagénomique en font de bons outils d’inventaire et de surveillance de virus potentiellement émergents.Dans un premier temps j’ai développé et validé un protocole expérimental de purification des viromes à ARN qui permettait le maintien de l’infectivité des particules virales. Ce protocole a ensuite été appliqué pour caractériser les communautés virales d’arthropodes hématophages et de prélèvements de faune sauvage. J’ai par la suite réalisé l’inventaire des communautés virales de viande de singe fumée illégalement importée en France et confisquée par les douanes, qui a révélé la présence de nombreux bactériophages, dont certains pourraient infecter des bactéries potentiellement pathogènes pour l’homme.Enfin j’ai caractérisé les communautés virales de culicoïdes collectés au Sénégal, ce qui a permis de mettre en évidence la présence de nombreux virus géants à ADN infectant les amibes. Le séquençage des viromes à ARN a quant à lui révélé la présence d'un certain nombre d'arbovirus qui pourraient constituer un risque d’émergence pour la santé humaine. Du fait de nombreux facteurs intrinsèques et extérieurs à l’agent infectieux, la prédiction des futures émergences de virus zoonotiques est très compliquée voire utopique, mais elle reste un challenge crucial et d’actualité. La stratégie de réalisation d’inventaires des communautés virales présentes dans les différents acteurs des cycles de transmission zoonotique est un premier pas indispensable dans la connaissance des risques potentiels d’émergence en population humaine
Zoonoses are responsible of more than two thirds of human viral infections. The development of high-throughput sequencing tools and their application in metagenomics allow inventorying the viral communities of various reservoirs in order to detect the emergence of viruses before their infection to humans. In this context, I characterized the viral communities of simian bushmeat illegally imported into France and of Culicoides biting midges, recognized vectors of several viruses of human and veterinary medicine importance. I have first developed a protocol for the purification of RNA viromes which allowed maintaining the infectivity of viral particles. This protocol was subsequently applied to characterize viral communities of bloodsucking arthropods and wildlife samples. In a second part I realized the inventory of viral communities of smoked simian bushmeat illegally imported into France and confiscated by the French customs. This study revealed the presence of a wide diversity of bacteriophages, in which some of them could infect bacteria potentially pathogenic for humans.Finally I characterized the viral communities of Culicoides biting midges collected in Senegal, which revealed the presence of sequences related to several giant DNA viruses infecting amoeba. Sequencing of the RNA virome revealed the presence of several arboviruses that could constitute a risk of emergence of zoonoses for humans.The prediction of future emerging zoonotic viruses is very difficult, if not impossible. However the characterization of viral communities present in the different actors of zoonotic transmission cycle is a first step to evaluate potential risks of transmission to humans

碩士
國立彰化師範大學
生物學系
97
Abstract Forcipomyia taiwana (Diptera: Ceratopogonidae ) is a blood sucking insect. Female midges were trapped by the human shank as the bait in Dakeng, Taichung County. After the female sucks full blood, and brought to laboratory and carried on the experiment. The experiment of the Forcipomyia taiwana ovipositional preference uses the algae which are Chlorella sp., Tetrachlorella sp., Chaetomorpha sp. of the green algae, Gloeothece sp. of the blue green algae, and the mix blue green algae (55 % Anabaena sp., 20 % Oscillatoria sp., 10 % Schizothrix sp. and 15 % Gloeocapsa sp.). The results showed that the amount of the accumulative laying eggs of the female midge will reach the peak in 48-54 hours after sucking the full blood. The female midge laid 4.3 eggs to an average on each egg mass, including about 1-6 eggs. The female midge spawned on different algae has significantly different (p<0.005), when the egg mass contained more than 5 eggs will aggregate significantly (p<0.005). The ovipositional preference of female midge on algae, especially on the number of eggs, was the mix blue green algae, Chaetomorpha sp., Gloeothece sp., Chlorella sp. and Tetrachlorella sp., respectively. On the number of egg masses were the mix blue green algae, Gloeothece sp., Chaetomorpha sp., Chlorella sp. and Tetrachlorella sp., respectively. The second peak of reflection spectrum appeared in Chlorella sp. and Tetrachlorella sp. have the higher reflectional coefficient, 64.4 % and 50.0 %, respectively, and next on Chaetomorpha sp., Gloeothece sp. and mix blue-green algae. The female midge of Forcipomyia taiwana spawned on low reflectional coefficient algae, the mix blue-green algae, has the high number of eggs and egg masses, but on high reflectional coefficient algae, Chlorella sp. and Tetrachlorella sp., have less ones. Key Words: Forcipomyia taiwana、ovipositional preference、reflection spectrum

A biosystematic study of seven Afrotropical and two Oriental species of the Imicola species-group was undertaken; this group of Culicoides of the subgenus Avaritia includes C. imicola the most important vector of the viruses of bluetongue (BT) and African horsesickness (AHS) known in the Old World. Five African species are redescribed i.e. C. imicola, C. pseudopallidipennis, C. bolitinos, C. miombo and C. loxodontis. Two new species are described, and the extralimital C. brevitarsis and C. nudipalpis are discussed where relevant. These nine species comprise the Imicola group, one of 10 groups constituting the subgenus worldwide. Due to confusion in the literature, the Imicola group is redefined and distinguished from the Orientalis group (also redefined); 21 species of approximately 70 world species of Avaritia are reassigned to either of the two groups. A key to all nine known species of the lmicola group is given; shortcomings in the taxonomy of the Orientalis group are discussed. The adult morphology of both sexes of the nine Imicola group species was studied; this revealed deficiencies in the descriptive format currently used in taxonomic studies of world Culicoides. Accordingly, greater detail has been introduced into descriptions and includes the use of new character states. Methods for mounting Culicoides on glass slides are also improved to ensure more accurate observation and measurement of diagnostic taxonomic features; furthermore, the descriptions are based on long series of each sex. Illustrations were made from specimens mounted symmetrically, and no feature omitted from any bodypart illustrated. Certain aspects of the life-cycle of most species were investigated but especially those of C. bolitinos, C. loxodontis and C. kwagga. The immatures of these develop exclusively in the dung of the elephant, the buffalo, the blue wildebeest, both species of rhinoceros and Burchell's zebra. Two of these species, C. bolitinos and C. kwagga, have broadened their resource range as they can invade and mature in the dung of cattle and horses. This has obvious implications for the transmission of viruses, especially where indigenous herbivores are run with domesticated livestock. In an intensive two-year survey a comparison was made between the Culicoides fauna of a natural area, the Kruger National Park (KNP), and that found in areas changed by Man, namely livestock farms adjoining the KNP. This part of the eastern Transvaal lowveld is a main focus of AHS in South Africa. Light-trapping, rearing from dung, and pootering off live hosts, revealed that some species of the Imicola group are exclusively associated with certain herbivores; these include the elephant and the zebra which are suspected or proven reservoir hosts for AHS. The results thus throw further light on the epidemiology of this disease, and also show that Man plays a decisive role in determining the numbers, and distribution, of particular Imicola group species under certain conditions. In the case of C. imicola, the commonest and most widespread of all species, this range expansion, or establishment of foci, is due to man's maintenance of domesticated livestock in confined species, and especially where these are kept on irrigated pastures. However, the serendipitous discovery of a large imicola-free zone in South Africa indicates that edaphic conditions likely play an even more important role than Man and climate in determining the prevalence and abundance of C. imicola. This area is the sandy dune field west of Port Elizabeth and holds promise as a natural quarantine zone for the import and export of livestock. AFRIKAANS : 'n Biosistematiese ondersoek van sewe Afrotropiese en twee Orientale spesies van die Imicola groep is gedoen; by hierdie groep Culicoides van die subgenus Avaritia word C. imicola wat as die mees belangrikste vektor van bloutong-(BT) en perdesiekte (AHS) virus in die Ou Wereld beskou word, ingesluit. Vyf Afrika spesies, nl. C. imicola, C. pseudopallidipennis, C. bolitinos, C. miombo en C. loxodontis, is herbeskryf. Twee nuwe spesies is beskryf asook die suid-oos Asiese spesies C. brevitarsis en C. nudipalpis word bespreek waar van toepassing. Hierdie nege spesies vorm die Imicola groep, een van die 10 groepe waaruit die subgenus Avaritia wêreldwyd bestaan. As gevolg van verwarring in die literatuur is die Imicola groep hergedefinieer en geskei van die Orientalis groep ( ook hergedefinieer); 21 spesies van ongeveer 70 wereld spesies van Avaritia is heringedeel in die twee groepe. 'n Sleutel vir al nege wereld spesies van die Imicola groep, asook 'n verspreidingskaart vir elke spesie, word gegee. Tekortkominge in die taksonomie van die Orientalis groep word ook bespreek. Die volwasse morfologie van beide geslagte van die nege Imicola groep spesies is bestudeer; dit het gebreke in die formaat wat tans vir die taksonomiese beskrywing van wêreld Culicoides gebruik word aan die lig gebring. Gevolglik is daar meer data in die beskrywings, wat nuwe karakterkenmerke insluit. Die metode van die montering van Culicoides op glasplaatjies is ook verbeter om meer akkurate ondersoek en meting van die diagnostiese kenmerke te verseker; verder, is beskrywings gebaseer op lang reekse van elke geslag. Illustrasies is gemaak van voorbeelde wat simmetries gemonteer is en geen kenmerk is uitgelaat van enige gelllustreerde liggaamsdeel. Aspekte van die lewensiklus van die meeste van die spesies, veral C. bolitinos, C. loxodontis en C. kwagga is ondersoek. Die onvolwassenes van hierdie spesies ontwikkel slegs in die mis van olifante, buffels, wildebeeste, renosters en zebras. Twee van die spesies, C. bolitinos en C. kwagga, het hulle broeimediums van voorkeur vergroot en kan eiers lê en tot volwassenheid ontwikkel in die mis van beeste en perde. Dit het vanselfsprekende gevolge vir virusoordrag tussen inheemse herbivore en vee, veral in gebiede waar die twee groepe saamloop. In 'n intensiewe twee-jaar studie, is 'n vergelyking gemaak tussen die Culicoides fauna soos aangetref in 'n ongerepte deel van Afrika, die Kruger Nasionale Park (KNP), en in gebiede aangrensend aan die KNP wat deur die mens in veeplase omskep is. Die deel van die Oos Transvaalse laeveld is bekend as 'n perdesiekte "hotspot" in Suid-Afrika. Ligvalvangste, uitbroei van Culicoides uit mis en versameling vanaf lewendige gash ere het aangedui dat sekere spesies van die Imicola groep eksklusief met sekere herbivore geassosieeris; hierby ingesluit is die olifant en zebra wat onderskeidelik verdagte en bevestigde gashere van AHS is. Die resultate dra by tot die verklaring van die epidemiologie van hierdie siekte en wys ook dat die mens 'n beslissende rol speel in die vasstelling van die getalle sowel as die verspreiding van spesifieke spesies van die Imicola groep. In die geval van C. imicola, die mees algemeenste en wydverspreidste spesie, is die uitbreiding, of daarstelling van fokuspunte, te wyte aan die mens se instandhouding van vaste bloedbanke op besproeide weiding. Die toevallige ontdekking van 'n groot imicola-vry sone elders in Suid-Afrika dui daarop dat grondtipe moontlik 'n bepalende rol kan speel in die aanwesigheid en volopheid van C. imicola. Hierdie "skoon" area is die sandduine-veld wes van Port Elizabeth en lyk belowend as 'n natuurlike kwarantyn gebied vir die invoer en uitvoer van lewende hawe.
Dissertation (MSc)--University of Pretoria, 2013.
Animal and Wildlife Sciences
unrestricted

Six new species of Dasyhelea Kieffer, 1911 from Argentinean and Chilean Patagonia are described and illustrated based on adults. Five of these species do not belong to any of the recognized groups found in the Americas. The remaining one, Dasyhelea pabloi sp. nov. is a typical member of the leptobranchia group and is the first record of a species from this group found in the Neotropical region.

碩士
國立彰化師範大學
生物學系
99
Foricipomyia taiwana is a diurnal insect, population cycle of diurnal blood-sucking activity began to rise from the early morning, the peak at ten o’clock in the morning to noon and population size decreased significantly in evening. This research was studied by field investigation, recording the density of Forcipomyia taiwana in blood-sucking activity, and estimating the relationship between temperature, humidity and other meteorological factors with the density of Forcipomyia taiwana. We try to predict in advance of the arrival of the peak, and trap the female adults of Foricipomyia taiwana with human body trapping method as early as we can. Thus we can reduce the social costs of combating with this animal. The female adults of Forcipomyia taiwana start blood-sucking activity and harassment human after mating stage in their lives. At this stage, they seize opportunities to suck blood from human. EPA used to control the epidemic situation by spraying over the past, but it not only leads to limited effect, but also causes secondary pollution on the environment. In this paper, the human body trapping method was used to capture the female adults of Forcipomyia taiwana, and reduce the number of groups in selected test areas at Puli, Nantou County. Test is divided into two phases. At the first stage of the investigation we observe the population density in different seasons. At the second phase, we mass trapped females adults of Forcipomyia taiwana. As a result, the population size before and after treatment compared with the density of both shows that mass trapping female adults of Forcipomyia taiwana can effectively reduce their population density significantly.

博士
國立中興大學
昆蟲學系所
98
This research focused on the biting midge, Forcipomyia taiwana (Shiraki). It was divided into three parts which included breeding techniques, bloodsucking habit and the trap development. Breeding techniques: On the agar-based substrate larvae only spent 9.4 days to develop into pupae. And the survival rate for larvae-to-adults and pupae was high to 93%, and 97%, respectively. Experiments on the ovipositional preferences of F. taiwana showed that female adults deposited the largest number of eggs on the agar-based substrate. It was found that F. taiwana bred on any one of these five different algae could all complete the larval and pupal development. Based on the developmental time and the survival rate, Scenedesmus sp. was the best one among all kinds of the tested algae for this biting midge. Under the tested temperatures from 15℃ to 35℃, it was found that the most suitable environment for F. taiwana to survive and reproduce was at 30℃. As the results of different food quantity and choice of oviposition sites for female adults, it was found that above 90% of female adults preferred to lay in the Petri dishes with larval food (algae) and the more food (0.01–0.16g) we put, the more eggs female adults laid. Bloodsucking habit: This study investigated the relationship between the bloodsucking activity of the midges and meteorological factors. Relative humidity was the major factor to influence the bloodsucking activity at the places with shelters, such as in the bamboo field, at the hut made of bamboo and at the site under the archway outside the bamboo field. The ultraviolet light and the air temperature had a positive correlation with the bloodsucking activity in the bamboo field and at the hut in the bamboo field, respectively. In terms of action spectrum, both males and females were more sensitive to ultraviolet light than to visible light, especially the spectral region between 330 nm and 340 nm for females and 370 nm for males, indicating that male and female adults preferred the ultraviolet light with different spectra individually. Trap development: The results showed that the LEDs were better than the fluorescent light for attracting biting midges. In this test the more attractive release rate of 1-octen-3-ol, lactic acid and acetone to this pest was found to be 0.7 mg/h, 0.2 mg/h, and 4.8 mg/h, respectively. Octenol was the most attractive to F. taiwana followed by lactic acid and acetone. However, the carbon dioxide with 250 ml/min for four release rates was the most attractive to this biting midge. The effectiveness of the trap with the combination of CO2 (release rates of 250 ml/min) + octenol (0.7 mg/h ) + blue light ( λmax = 405 nm) was the best among four combinations for this pest.

碩士
國立中興大學
昆蟲學系所
98
The field density of the biting midge, Forcipomyia taiwana (Shiraki) (Diptera: Ceratopogonidae), was studied from Nov. 2009 to Apr. 2010 in Taichung, Taiwan. The result show that F. taiwana density was low in winter and rapidly increased during spring. The reproductive potential of female midges shows, however, no difference between sampling dates. It indicates that the preadult mortality might be one of the major factors of population density in the field. The demography of the biting midge was studied based on the age-stage, two-sex life table for cohorts fed on human blood and mouse blood. The intrinsic rate of increase for cohort fed on human blood was 0.0966 d-1, and 0.1296 d-1 for that fed on mouse blood. The net reproductive rate and the mean generation time was 14.6 offspring and 27.9 d for cohort fed on human blood, and 19.5 offspring and 23 d for cohort fed on mouse blood, respectively. The population projection shows that midge population suffers high preadult mortality in the field.

碩士
國立中興大學
昆蟲學系
93
Effects of food and humidity on the development of Forcipomyia taiwana (Diptera: Ceratopogonidae) Pei-Shiou Chen Abstract The biting midge, Forcipomyia (Lasiohelea) taiwana (Shiraki) (Diptera: Ceratopogonidae) is a human-biting pest. The larvae were fed on three different strains (CH1, CH2 and drought-resistant) of algae (Anabaena sp.) after hatching. Among three different strains, when fed on Anabaena sp. drought-resistant strain, the developmental duration required 21 days. When the larvae were fed on the other algae, they needed 17 days to develop to adult. When the larvae were placed on agar with moss, they often died without further development. Among three different larval densities tested, the developmental duration with 20 larvae was shortest. Females were collected from field and kept at a photoperiod of 12 : 12 (L : D) h or 12 : 12 (D : L). After incubating for four days with blood meal, the abundance of eggs laid was not significantly different between two photoperiod regimes. The LT50 values of first to fourth instar under submergence test were 4.3 days to 16 days. The mortality was significantly different among different larval instars. The LT50 of pupa under submergence test was 30.6 h. The emergence rate was significantly different among various submergence times. While the relative humidity increased from 50.5% to 100%, the emergence rate of F. taiwana was increased significantly.

African horse sickness virus causes a non-contagious, infectious disease of equids. It is epizootic to sub-Saharan Africa and parts of the Middle East. The epizootics caused by the virus have caused widespread devastation amongst equids worldwide. Fortunately no epizootic has lasted more than 5 years outside of sub- Saharan Africa. It is vectored by species of Culicoides midges (Diptera: Ceratopogonidae) and most importantly by the two Avarita species of C. imicola Keiffer and C. bolitinos Meiswinkel. The literature pertaining to the study and research of the virus, the disease and the vectors is reviewed. Models allowing prediction of future possible outbreaks as well as details of control strategies and findings of researchers are presented and discussed. The virus needs a long term reservoir host in which to overwinter and various theories are discussed. Control measures in South Africa are suggested so that outbreaks of the disease can be reduced.
Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermarizburg, 2008.

Tese de mestrado em Biologia Humana e Ambiente, Universidade de Lisboa, Faculdade de Ciências, 2020
De acordo com a Organização Mundial de Saúde, as doenças transmitidas por vetores representam mais de 17% de todas as doenças infeciosas no mundo, originadas por agentes patogénicos como vírus, bactérias e parasitas. A distribuição destes agentes tem vindo a ser afetada pela recente combinação de fatores como a globalização comercial, o transporte rápido e acessível e o menor rigor das regulamentações internacionais de saúde em viagens, causando o aumento da disseminação de parasitas e de vetores para novas áreas geográficas, colocando populações de humanos e outros animais, anteriormente não expostos, em risco de infeção. Os insetos do género Culicoides (Diptera: Ceratopogonidae) possuem uma grande importância na saúde animal e humana, uma vez que as fêmeas hematófagas são vetores de vários agentes patogénicos, como vírus (Língua Azul, Peste Equina Africana e Schmallenberg), filarídeos (Mansonella spp.), entre outros. Por sua vez, as fêmeas do género Culicoides são responsáveis por elevadas perdas económicas a nível mundial. Um conhecimento mais aprofundado da fauna de Culicoides presente em cada país é necessário, assim como as suas preferências ecológicas, de modo a que medidas possam ser aplicadas de modo mais eficaz e sejam criadas diferentes estratégias de controlo. A doença da Língua Azul é uma doença viral, não contagiosa que afeta ruminantes domésticos e silváticos, que foi reconhecida e descrita há mais de 230 anos na África do Sul. O vírus da doença da Língua Azul era transmitido por vetores endémicos de regiões tropicais e temperadas até meados dos anos 90 do século XX. No entanto, nos últimos anos têm ocorrido mudanças drásticas na distribuição mundial destes vetores portadores de vírus, particularmente na Europa desde 1998, onde surgiu na região sul do continente, muito possivelmente devido à introdução de hospedeiros ou produtos infetados, provenientes de regiões endémicas, ou às alterações climáticas que permitiram a persistência do vírus durante o inverno e a expansão para norte de Culicoides imicola Kieffer, 1913, o principal vetor da doença. Além disso, desde 2006, novos casos da doença foram reportados em regiões mais a norte da Europa, onde a espécie Culicoides imicola não existe, indicando que outras espécies, como as pertencentes ao grupo Obsoletus, têm competência vetorial. No entanto, existem poucos estudos sobre a associação entre Culicoides e animais silváticos e, por isso, o papel destas espécies devia ser investigado, já que a alimentação oportunista em diferentes hospedeiros pode facilitar a transferência de vírus entre animais silváticos, domésticos e até em humanos. Neste projeto, foram realizadas capturas de espécies perto de animais silváticos (girafas, zebras e aves) no Jardim Zoológico de Lisboa, entre maio de 2018 e setembro de 2019, e perto de bovinos domésticos da Faculdade de Medicina Veterinária da Universidade de Lisboa, entre junho e setembro de 2019 (excluindo o mês de agosto por ser período de férias de verão). Além dessas capturas, foram selecionadas capturas mais antigas de uma quinta em Leiria, realizadas em 2010 durante o Plano Entomológico de Vigilância Nacional para a doença da Língua Azul em Portugal (2005-2013), que foram utilizadas para comparar capturas mais antigas com as mais recentes. Como o conhecimento dos padrões de alimentação das diferentes espécies de Culicoides é essencial para o conhecimento da sua capacidade vetorial e como a determinação das preferências de hospedeiros clarifica o papel destas espécies na epidemiologia das diferentes doenças, os objetivos específicos deste trabalho são compreender melhor a distribuição destas espécies de Culicoides entre: i) animais silváticos e bovinos domésticos; ii) três animais silváticos diferentes: girafas, zebras e aves; iii) bovinos domésticos: Leiria, capturas mais antigas e Lisboa, capturas mais recentes e iv) detetar e identificar através de análise molecular as anomalias morfológicas em Culicoides do grupo Obsoletus. Para tal, os Culicoides foram capturados com recurso a armadilhas luminosas do tipo CDC, que são utilizadas durante o período noturno. Os insetos capturados foram analisados morfologicamente, mostrando a variação das espécies de Culicoides nestes locais. As capturas realizadas nos meses entre junho, julho e setembro de 2019 no Jardim Zoológico de Lisboa e na Faculdade de Medicina Veterinária foram comparadas e, os resultados demonstraram que de um total de 135 exemplares capturados perto de animais silváticos, foram capturadas oito espécies diferentes. Por outro lado, foram identificadas, perto de bovinos domésticos, sete espécies de Culicoides de um total de 158 exemplares. Destas espécies que foram identificadas nos dois locais é de notar que a maior abundância vai para os principais vetores da doença na Europa, C. imicola e espécies pertencentes ao grupo Obsoletus (C. obsoletus e C. scoticus). Este resultado pode suportar o papel putativo destas espécies como ponte de transmissão de arbovírus entre animais silváticos e ruminantes domésticos que, consequentemente, uma vez introduzidos e sob as condições ideais, se disseminarão rapidamente por grandes regiões da Europa. Em Portugal, este é o primeiro estudo realizado da fauna de Culicoides capturados perto de animais silváticos e a sua comparação com capturas realizadas perto de ruminantes domésticos. A monitorização da fauna de Culicoides em ambiente de zoo é importante para evitar o potencial risco de introdução de doenças vindas de animais importados. Neste contexto, as capturas foram realizadas no Jardim Zoológico de Lisboa entre maio de 2018 e setembro de 2019, de onde foram coletados 1088 exemplares, dos quais 71% capturados perto de girafas, 28% perto de zebras e apenas 1% perto de aves. Uma vez mais, as espécies mais capturadas foram C. imicola e aquelas pertencentes ao grupo Obsoletus. Em relação à comparação dos resultados obtidos, com um distanciamento de nove anos nas explorações de gado de Leiria em 2010 e de Lisboa em 2019, é importante verificar que ocorreu uma queda acentuada no número de exemplares capturados. Esta redução, provavelmente, pode ter ocorrido devido ao ano quente e seco que se fez sentir em 2019 que dificultou a criação de condições favoráveis para o surgimento de Culicoides. Além disso, a composição de Culicoides nestas duas capturas revelou que as espécies pertencentes ao grupo Obsoletus foram as mais capturadas, representando 71% das capturas em 2010 e 81% das capturas em 2019. A segunda espécie mais capturada em ambos os anos foi C. punctatus, sem diferenças significativas na composição de ambas as capturas. Além disso, nestes dois anos, junho foi o mês com mais exemplares capturados, como esperado, tendo em conta os padrões da dinâmica sazonal destas espécies. Tendo em conta que os indivíduos pertencentes ao grupo Obsoletus são muito semelhantes, a sua identificação apenas pelo padrão da asa não seria possível. Portanto, para separar estas espécies dentro do grupo Obsoletus é sempre necessário analisar as várias estruturas morfológicas e, por isso, foram detetadas alterações morfológicas. Estas alterações morfológicas podem dever-se ao cruzamento de espécies dentro deste grupo que são muito próximas e/ou ao facto de, dependendo da espécie, de depositarem entre 10 e 675 ovos, o que pode gerar a probabilidade de ocorrerem erros genéticos que podem causar estas malformações anatómicas. Neste projeto podem ser encontradas e descritas características morfológicas anormais, como é o caso de segmentos do palpo fundidos, exemplares com uma ou três espermatecas, em vez de duas funcionais e uma rudimentar. Com este trabalho é reportado pela primeira vez em Portugal um exemplar de C. obsoletusinfértil que possuía apenas uma espermateca e esta não funcional. De forma a compreender a posição taxonómica destes espécimes com malformações anatómicas, foi realizada uma análise molecular, mas depois de várias tentativas não foram obtidos resultados. No total das capturas realizadas neste projeto, as espécies mais prevalentes são as principais vetoras da doença da Língua Azul na Europa, C. imicola e espécies pertencentes ao grupo Obsoletus (C. obsoletus (Meigen, 1818) e C. scoticus (Downes & Kettle, 1952). A distribuição e abundância destas espécies é afetada por fatores abióticos, como a temperatura, a exposição ao vento, entre outros fatores como a proximidade, exposição, densidade e tamanho de potenciais hospedeiros à armadilha, bem como a disponibilidade de habitats ideais ao crescimento das larvas de Culicoides. Existe uma grande preocupação quanto às alterações climáticas e o potencial de permitirem a expansão de vários agentes patogénicos transmitidos por vetores, uma vez que estes são muito sensíveis ao clima. Os resultados deste projeto evidenciam e sustentam que, tendo em conta o impacto económico associado aos agentes patogénicos que transmitem e a constante expansão territorial de novos vetores, é importante realizar estudos entomológicos dos insetos do género Culicoides. Concluindo, é importante ter conhecimento da epidemiologia de uma doença de forma a poder criar as medidas corretas de prevenção, e, além disso, o aumento do conhecimento nesta área permite a consciencialização do perigo deste tipo de doenças. Um conhecimento mais profundo na fauna de Culicoides presente em cada região e as suas preferências quanto a hospedeiros e ambiente é exigida para criar estratégias de controlo que sejam mais eficazmente aplicadas. Para este efeito, deveriam ser realizadas mais monitorizações ao longo do país e estudos onde a variação temporal das capturas, seja semanal ou ao longo de todo o ano, bem como a realização de estudos para se conseguir compreender melhor as anomalias morfológicas encontradas e a sua posição taxonómica, quando comparados com espécies já conhecidas.
Culicoides biting midges (Diptera: Ceratopogonidae) have a major importance in animal and human health since they are vectors of several pathogens, like viruses (Bluetongue, African Horse Sickness and Schmallenberg), filarial nematodes (Mansonella spp.), among others. Bluetongue is an arthropod-transmitted viral disease of domestic and sylvatic ruminants that was recognized and described more than 230 years ago in southern Africa. Bluetongue disease was enzootic throughout tropical and temperate regions of the world until 1998. Then, Bluetongue disease expanded to southern European countries and this spread has been driven by global warming, introduction of infected hosts or infected products from endemic regions that have allowed increased virus persistence, maintenance during the winter and the northward expansion of Culicoides imicola Kieffer, 1913, the main bluetongue virus vector. Female biting midges are responsible for huge economical losses worldwide and, since 2006, new Bluetongue-serotypes have also been reported from countries in Northern and Western Europe, where C. imicola is absent. However, little is known about Culicoides species associated with natural environments, and their role as vector species in wildlife should be investigated since opportunistic host feeding may facilitate virus transfer between wild and domestic hosts and even to humans. The present work is based on Culicoides species captured near sylvatic animals (giraffes, zebras and birds) from Lisbon Zoo, between May 2018 and September 2019, and domestic cattle (cows) from Faculty of Veterinary Medicine, University of Lisbon captured between June 2019 and September 2019 (excluding August because the holiday period). Plus, older collections of cattle from a farm in Leiria District performed in 2010, during the National Entomologic Surveillance Program for Bluetongue disease in mainland Portugal (2005-2013), were used to compare those captured species with the most recent ones. Culicoides were collected with miniature CDC light traps. Then, biting midges were identified by morphological features, in order to understand the variation of Culicoides species in these collection sites. Since knowledge of the blood-feeding behaviour of Culicoides midges is essential in assessing their vectorial competence and determining host preferences clarifies the roles of these species in the epidemiology of different diseases, the main aim of this work was to better understand how the distribution of Culicoides species is between sylvatic animals and domestic cattle. Specific goals include the comparison of Culicoides species captured near: i) sylvatic animals and domestic cattle; ii) three different sylvatic animals: giraffes, zebras and birds; iii) domestic cattle from: Leiria and Lisbon in two different time points and iv) detection and molecular identification of morphological anomalies in Culicoides from Obsoletus group. The captures made between June and September 2019 (excluding August) were used to compare the captures made in Lisbon Zoo and in the Faculty of Veterinary Medicine, where from a total of 135 biting midges collected near sylvatic animals, eight Culicoides species were identified. On the other hand, from 158 Culicoides female specimens collected near domestic cattle, seven were identified. Also, this study showed the abundance of the main BTV vectors in Europe, C. imicola and Obsoletus group species, near sylvatic animals and domestic ruminants. That would support their putative role as bridge vectors for the transmission of the arboviruses between sylvatic animals and domestic cattle and once introduced, consequently, they will spread rapidly over large regions of Europe when appropriate environmental conditions and hosts are present. This is the first study in Portugal of Culicoides fauna captured near sylvatic animals and their comparison with those captured near domestic cattle. The monitorization of Culicoides biting midges in zoo environment is important to avoid potential risk of outbreaks since imported animals could be reservoirs of diseases and zoo is a place where a large number of potential vectors may be present. In Lisbon Zoo, from a total of 1088 Culicoides biting midges, the biggest amount of Culicoides was collected near giraffes, representing 71% of all captures, followed by zebras with 28% and birds with 1%. C. imicola and species from Obsoletus group were the most captured ones. When comparing the 9-year period in farm environments from Leiria in 2010 and Lisbon in 2019, it is important to notice that a drop in captured Culicoides species occurred. Probably this drop happened because of the hot and dry year, that may cause a deficit in the occurrence of favourable conditions for their emergence. However, Obsoletus group species represented in both years the most captured species, with 71% and 81% of all captures in 2010 and 2019, respectively. C. punctatus represented the second most captured species in both years, without significative differences in captures composition. Besides that, in both years, June was clearly the month with more captured specimens, following the characteristic pattern of seasonal dynamic of those species. Since species inside Obsoletus group are very similar, their identification is not possible only by their wing pattern. So, in order to be identified they need to be dissected into different body parts. Through the detailed observation of these midges, it was possible to detect abnormal morphological structures. These abnormal structures may be due to the crossbreeding of close related species or the fact that they deposit 10 to 675 eggs, depending on species, that resulted in genetic errors that could cause morphological modifications. This project gives essential information concerning these aberrant characteristics, including a specimen with only one non-functional spermathecae in C. obsoletus, which was never reported before to the best of our knowledge. Furthermore, a molecular analysis of these specimens was made to understand their taxonomical position but, after several attempts, no results were obtained. Known BTV vectors in Europe, as C. imicola and species from Obsoletus group (C. obsoletus (Meigen, 1818) and C. scoticus (Downes & Kettle, 1952), were the most captured species during this project. Their distribution and abundance are affected by abiotic factors, such as climate, temperature, wind exposure, soil type, surrounding vegetation and other factors, as the potential host proximity and exposure, density and size of hosts nearby the trap and the availability of larval habitats close to the traps. There is a big concern that climate change will lead to expansion of vector-borne diseases, as they are more sensitive to those factors, since these can affect tens of thousands of farms with high financial costs to farmers and countries and causing the death of millions of animals. Thus, such expansion may threaten human health and food security via effects on animal and crop health. Also, this project gives essential information concerning morphological modifications that can be observed in Culicoides biting midges, some of them referred for the first time. Understanding the epidemiology of a disease allow us to take the correct measures to predict and prevent it, as well as enhancing our knowledge about the emergence of other vector-borne pathogens. A deeper knowledge of Culicoides fauna present in each region and their ecological preferences is required, so different control strategies can be applied efficiently.

碩士
中興大學
昆蟲學系所
95
The gonotrophic cycle of the blood-feeding females of Forcipomyia taiwana (Shiraki) were complete at 15℃, 20℃, 25℃, 30℃, and 35℃. Egg developmental rate in the gravid female is increase with temperature rising, and the oviposition rate of gravid female is significantly high at 25℃ and 30℃. Normally, there are circadian rhythm phenomenon in the ovipositiion of gravid female under 25℃, the higher peak of eggs laid are located at 12:00 to 18:00. The oviposition of gravid female F. taiwana is effected by the temperature, light, and the food of larva, and the light factor is most important that affects the oviposition. Pyriproxyfen is the effectiveness insect growth regulator (IGR) on the susceptibility of 2nd and 4th larva of F. taiwana among the three tested IGRs, Pyriproxyfen, Flufenoxuron, and Chlorofluazuron. There are 125 village or towns in 13 counties occurs F. taiwana under the distribution investigation during August to October, 2006. A highest collected record is at the 900 meters high upper sea in Nantou area. Seasonal dynamic of the population of F. taiwana was surveyed at two countries area, Shioufeng in Lugu village and Hsiolin in Jhushan township during May, 2005 to May 2007. These investigations show the low population density of F. taiwana in winter and early Spring seasons, and the density were increasing with temperature higher, than the highest population density was located at Summer seasons, in July and August. The dynamic of F. taiwana population density were affected by climate fluctuation. By using the correlation and multiple regression analysis, the temperature, relative humidity, and raining period are the most correlation meteorological factors with population density of F. taiwana, especially two weeks before the midge collected.

碩士
國立中興大學
昆蟲學系所
99
The density of female adults of the biting midge, Forcipomyia taiwana (Shiraki) (Diptera: Ceratopogonidae) was monitored in Dakeng Scenic Area and the campus of National Chung Hsing University, Taichung, Taiwan. On both study sites, the density of female adults was higher during the period from March to October than that of other months. The mean fecundity of females collected from field is 42.9 eggs/female with a range from 24.0 to 64.8. The average egg hatch rate is 59.4% ranging from 46.5% to 71.5%. Based on the age-stage, two-sex life table theory, life tables of F. taiwana reared on green algae, Chlorella sp. of different concentrations were collected. When larvae were supplied with concentrated algae, the intrinsic rate of increase and mean generation time were 0.1209 d-1 and 18.3 d, respectively. Life tables collected at different algae concentrations showed that 1.5 billion cells/ml and 2.0 billion cells/ml resulted in higher intrinsic rates of increase.

碩士
國立臺灣大學
環境衛生研究所
104
Mosquito (Culicidae) are the dominant vectors of pathogens that cause infectious disease such as malaria, yellow fever, and especially dengue in the world. Also, Forcipomyia taiwana is a diurnal, tiny, blood-sucking indigenous midge that solely feeds on human and distributes island-wide in Taiwan, either. Individuals expose to midge bites will develop intense pruritus and swelling immediately or delayed allergic responses, causing declination of quality of people’s daily life. Therefore, preventing biting of mosquitoes and nuisance of biting midges by trapping/repelling adults and eliminating larvae is important. This study aims to utilize push-pull strategies to develop an eco-friendly integrative control method, using light-based attraction tool for baiting adults of F. taiwana and to investigate the larvicidal activity of essential oils (EO) against F. taiwana larva, repellent activity of EO against F. taiwana adult and mosquitoes. The first results suggested that visible blue light (wavelength 400~450nm) attracted three times more F. taiwana than UVA light (wavelength 352 nm) and UVB light (wavelength 306 nm) in a small scale experiment, the attraction rate are 70.7%, 18.3% and 16.0% separately. However, as in a field study, visible red light (wavelength 630nm) has the highest attraction rate (7.4%) than visible blue light (1.8%), visible green light (2.2%), full spectra light (2.8%) and UV light (0%). Second, six kinds of EOs from leaves of plants (Cryptomeria japonica, Cinnamomun osmophloeum, Clausena excavate, Gaultheria cumingiana, Cinnamomun brevipedunculatum and Melaleuca alternifolia) were applied with regard to larvicidal activity of F. taiwana larva. The results showed that larvicidal activity of EO is ranged as below: C. osmophloeum = C. excavate > M. alternifolia > G. cumingiana > C. japonica > C. brevipedunculatum. To be more specific, C. osmophloeum completely suppressed larvae activities with a concentration of 7.9 μg/cm2. EO from C. excavate, M.   alternifolia, G. cumingiana and C. brevipedunculatum showed larvicidal activity with 0%, 20%, 50% and 86.7% of larvae survived with concentration of 15.8 μg/cm2. Third, the repellency of C. japonica, C. osmophloeum and C. excavate against F. taiwana were evaluated at EO concentration 20%, 10%, 5%, 2.5%, 1.75%. 15% DEET is positive control. The results reveal that the average repellency of C. japonica ranged as 40.9±15%, 59.1±13.2%, 55.1±5.9%, 47±4.9% and 37.9±6.5% in 120 mins, which are all higher than 15% DEET’s repellency: 24.2±24.8%. Also, C. osmophloeum exhibited the best repellent activity ranged as 66.1±6.7%, 68.9±5.3%, 51.5±5.5%, 26.7±29.9% and 41.8±11.1% in 120 min, which are all higher than 15% DEET’s repellency: 20.5±11.8%, either. Forth, the repellency of C. japonica, C. osmophloeum and C. excavate against Aedes aegypti, Aedes albopictus, Culex tritaeniorhynchus and Armigeres subalbatus were evaluated at EO concentration 20%, 10%, 5%, 2.5%, 1.75%. 15% DEET is positive control. The ANOVA statistics results showed that each kind of EO had repellency specificity toward different species of mosquitoes. For example, C. japonica had well repellency against Ae. aegypti, Cx. tritaeniorhynchus and Ar. subalbatus, yet it had repellency -2.7±22.4% and -7.4±23.7% toward Ae. albopictus at lower concentration 2.5% and 1.75% respectively, showed that it possessed attraction activity. Moreover, with Fisher least significant difference (LSD) Test, it showed that C. osmophloeum and C. japonica had higher repellency than C. excavate at high concentration 20% and 10%; besides, C. osmophloeum still exhibited highest repellency at low concentration 5%, 2.5% and 1.75%. Based on the above results, the EO of C. osmophloeum showed the greatest repellency. This study utilize the integrative concept of Push-Pull Control Strategy to determine the attraction rate of light trap toward F. taiwana adult in small scale and field study, the larvicidal activity of essential oils against F. taiwana larva, and the repellency of essential oils against F. taiwana adult and mosquito, providing an sustainable, feasible, and eco-friendly integrated methods for pest control. In the future, we expect that our promising findings can be widely applied into personal protection and outdoor environment control, in order to decrease the transmission of mosquito-borne diseases.