RESUMO
BACKGROUND: In the context of the ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, the supply of personal protective equipment remains under severe strain. To address this issue, re-use of surgical face masks and filtering facepiece respirators has been recommended; prior decontamination is paramount to their re-use. AIM: We aim to provide information on the effects of three decontamination procedures on porcine respiratory coronavirus (PRCV)-contaminated masks and respirators, presenting a stable model for infectious coronavirus decontamination of these typically single-use-only products. METHODS: Surgical masks and filtering facepiece respirator coupons and straps were inoculated with infectious PRCV and submitted to three decontamination treatments, ultraviolet (UV) irradiation, vaporized H2O2, and dry heat treatment. Viruses were recovered from sample materials and viral titres were measured in swine testicle cells. FINDINGS: UV irradiation, vaporized H2O2 and dry heat reduced infectious PRCV by more than three orders of magnitude on mask and respirator coupons and rendered it undetectable in all decontamination assays. CONCLUSION: This is the first description of stable disinfection of face masks and filtering facepiece respirators contaminated with an infectious SARS-CoV-2 surrogate using UV irradiation, vaporized H2O2 and dry heat treatment. The three methods permit demonstration of a loss of infectivity by more than three orders of magnitude of an infectious coronavirus in line with the United States Food and Drug Administration policy regarding face masks and respirators. It presents advantages of uncomplicated manipulation and utilization in a BSL2 facility, therefore being easily adaptable to other respirator and mask types.
Assuntos
Infecções por Coronavirus/prevenção & controle , Descontaminação/normas , Reutilização de Equipamento/normas , Temperatura Alta , Peróxido de Hidrogênio/normas , Dispositivos de Proteção Respiratória/virologia , Equipamentos Cirúrgicos/normas , Equipamentos Cirúrgicos/virologia , Raios Ultravioleta , Guias como Assunto , HumanosRESUMO
Insects are seen as a solution to the increasing demand for protein sources for food. However, entomophagy has unfortunately been linked to allergic reactions in Europe with people with professional contacts. As mealworms (Tenebrio molitor) and crickets (Acheta domesticus) have recently become commercially available (both whole or in food formulation) in several European countries, this research assessed the cross allergenicity of arginine kinase (AK). Based on the collection of sera from a entomology laboratory staff, oven cooked insects but also purified AK fractions were tested. Immunoblotting against the protein extracts revealed different Immunoglobulin E reactivity of sera according to the insect target species: two bands (40 and 14â¯kDa) for crickets and a pattern including light responses at 17, 25 and 37â¯kDa for mealworms. Focusing on AK, low specific allergenicity was here illustrated and discussed in relation to the development of a safe edible insect consumption by humans.
Assuntos
Alérgenos/imunologia , Arginina Quinase/imunologia , Gryllidae/imunologia , Proteínas de Insetos/imunologia , Tenebrio/imunologia , Adulto , Animais , Culinária , Reações Cruzadas , Eletroforese , Feminino , Hipersensibilidade Alimentar/imunologia , Humanos , Imunoglobulina E/imunologia , Proteínas de Insetos/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Saliva is a critical biochemical interface between aphids and their host plants; however, the biochemical nature and physiological functions of aphid saliva proteins are not fully elucidated. In this study we used a multidisciplinary proteomics approach combining liquid chromatography-electrospray ionization tandem mass spectrometry and two-dimensional differential in-gel electrophoresis/matrix-assisted laser desorption/ionization time-of-flight/mass spectrometry to compare the salivary proteins from three aphid species including Acyrthosiphon pisum, Megoura viciae and Myzus persicae. Comparative analyses revealed variability among aphid salivary proteomes. Among the proteins that varied, 22% were related to DNA-binding, 19% were related to GTP-binding, and 19% had oxidoreductase activity. In addition, we identified a peroxiredoxin enzyme and an ATP-binding protein that may be involved in the modulation of plant defences. Knowledge of salivary components and how they vary among aphid species may reveal how aphids target plant processes and how the aphid and host plant interact.
Assuntos
Afídeos/genética , Proteínas de Insetos/biossíntese , Proteínas e Peptídeos Salivares/genética , Sequência de Aminoácidos , Animais , Proteínas de Insetos/genética , Espectrometria de Massas , Proteoma , Proteômica/métodos , Proteínas e Peptídeos Salivares/biossínteseRESUMO
Chemoreception plays an important role in mediating a diverse range of behaviours, including predation and food selection. In the present study, we combined anatomical observations, electrophysiology and proteomics to investigate sensilla that mediate chemoreception on the antenna and the legs of Tribolium. Scanning electron microscopy was used to differentiate the coxal and trochanteral segments of the pro-, meso- and metathoracic legs by the presence of sensilla trichoidea and chaetica, while the antennae were covered with five types of sensilla (chaetica, basiconica, trichoidea, squamiformia and coeloconica). Antenna morphology and ultrastructure were similar in both sexes. Electrophysiological recordings allowed us to characterize a row of small sensilla basiconica on the terminal segment of the antenna as taste receptors, responding to sucrose and NaCl. Proteomics investigations of antennae and legs yielded several proteins with specific interest for those involved in chemoreception. Odorant-binding proteins were antenna-specific, while chemosensory proteins were detected in both tissues.
Assuntos
Antenas de Artrópodes/metabolismo , Proteômica/métodos , Sensilas/metabolismo , Paladar/genética , Animais , Antenas de Artrópodes/ultraestrutura , Células Quimiorreceptoras/metabolismo , Proteínas de Insetos , Microscopia Eletrônica de Varredura , Sensilas/ultraestrutura , Tribolium/genéticaRESUMO
The inventory of pests occurring on Dacryodes edulis (Burseraceae) was carried out in rural areas in Gabon during 2009 and 2010. Yellow traps and visual observations were used to record weekly pests during the tree flowering stage, in five villages. Catches from yellow traps rose to 7,296 and 1,722 insect pests in 2009 and 2010, respectively, whereas records from visual observations corresponded to 1,812 and 171 insect pests in 2009 and 2010, respectively. During both years, abundance from traps and visual monitoring was significantly different between sampling sites (p < 0.05). The difference in pests' diversity between sampling sites was not significant (p > 0.05) according to traps, but significant (p ≤ 0.04) according to visual observations in 2010. Mecocorynus loripes Chevrolat (Coleoptera: Cucurlionidae) attacked the stem of D. edulis, while Oligotrophus sp. (Diptera: Cecidomyiidae), Pseudophacopteron serrifer Malenovsky and Burckhardt (Hemiptera: Phacopteronidae), and Selenothrips rubrocinctus Giard (Thysanopera: Thripidae) attacked leaves. Pseudonoorda edulis Maes and Poligui (Lepidoptera: Crambidae) and Lobesia aeolopa Meyrick (Lepidoptera: Tortricidae) infested fruits and inflorescences, respectively. These insects are specifically linked to plant patterns, and their identification provided the first basic information for developing suitable strategies to control pests of D. edulis in Gabon, as well as in neighboring central African countries.
Assuntos
Burseraceae , Besouros , Hemípteros , Controle de Insetos , Animais , Frutas , GabãoRESUMO
The different steps of the foraging process of wireworms (Coleoptera: Elateridae) would be better understood if accurate and holistic information regarding the role of plant-produced chemicals constituting their environment were available. Volatile organic compounds (VOC) play important roles in the interactions between plants and insects in many ecosystems, whether they take place aboveground or below-ground. The roles of VOC are still relatively unknown for wireworms, and deserve attention. Here, we performed three experimentations with barley roots as baits. In the two first, we assessed the effect of chopped roots and fungus infected roots on the orientation of wireworms. In the third experiment, the larvae were confronted to both healthy and fungus infected roots. We discuss the results in terms of suitability of the olfactometers we designed for the investigation of olfaction in wireworms, and we provide suggestions to improve their use.
Assuntos
Besouros/fisiologia , Hordeum/parasitologia , Doenças das Plantas/parasitologia , Animais , Comportamento Animal , Bioensaio , Fungos/fisiologia , Hordeum/microbiologia , Hordeum/fisiologia , Interações Hospedeiro-Parasita , Doenças das Plantas/microbiologia , Raízes de Plantas/microbiologia , OlfatoAssuntos
Mel/análise , Compostos Orgânicos Voláteis/química , Animais , Abelhas , Carboidratos/química , RomêniaRESUMO
Dr. Marcel Leclercq was a pioneer in the field of forensic entomology. He has provided his knowledge of insect biology to many forensic cases, and most of them have found the way to publication. Most of the articles he has written were focused on individual cases, and despite the abundance of entomoforensic investigations he conducted, no synthesis has been published. This article summarizes 36 yr of forensic entomological investigations in temperate Europe, mainly in Belgium. Dr. Leclercq's work includes 132 entomological cases involving 141 human corpses found in various death scenes. Under certain conditions, insect specimens found at death scene can provide information on when (postmortem interval estimation), where and how a person died. More or less 100 insect species associated with a dead body have been identified by Dr. Leclercq.
Assuntos
Entomologia/história , Ciências Forenses/história , Insetos/classificação , Animais , Bélgica , Cadáver , França , História do Século XX , História do Século XXI , HumanosAssuntos
Comportamento Animal/fisiologia , Besouros/fisiologia , Animais , Comportamento Animal/efeitos dos fármacos , Besouros/efeitos dos fármacos , Hidrocarbonetos/metabolismo , Hidrocarbonetos/farmacologia , Controle de Insetos/métodos , Espécies Introduzidas , Atividade Motora , Controle Biológico de Vetores , Estações do AnoRESUMO
To evaluate postmortem intervals (PMIs), one should take into account the determined age of necrophagous flies present on the cadaver. However, PMI determination needs further improvement, and rapid and accurate approaches have therefore to be developed. While previous studies have focussed on insect cuticular hydrocarbons, here we explore the volatile profile released by larvae and pupae of Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae). We monitored changes in volatile compounds daily, by headspace solid-phase microextraction, followed by gas chromatography-mass spectrometry. Branched and unbranched hydrocarbons, alcohols, esters and acids were identified, and the volatile profile was shown to vary, in both composition and quantity, with the age of the larva/pupa under investigation. We concluded, based on the analysis of the released volatile organic compounds, that it is possible to increase the accuracy of the estimated PMI, through improved estimation of the age of blowflies present on the cadaver.
Assuntos
Dípteros/química , Comportamento Alimentar , Compostos Orgânicos Voláteis/análise , Animais , Entomologia , Patologia Legal , Cromatografia Gasosa-Espectrometria de Massas , Larva/química , Mudanças Depois da Morte , Pupa/química , Microextração em Fase Sólida , SuínosAssuntos
Besouros/efeitos dos fármacos , Furanos/farmacologia , Hordeum/metabolismo , Alginatos/química , Animais , Besouros/crescimento & desenvolvimento , Besouros/fisiologia , Comportamento Alimentar , Cromatografia Gasosa-Espectrometria de Massas , Controle de Insetos , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/fisiologia , Microesferas , Raízes de Plantas/metabolismo , Microextração em Fase SólidaRESUMO
The cuticular hydrocarbons of insects are species-specific and often function as semiochemicals. The activity of Tribolium brevicornis cuticular hydrocarbons as feeding deterrents that ostensibly function to prevent pupal cannibalism and predation was evaluated. The cuticular hydrocarbons of T. brevicornis pupae were characterized and flour disk bioassays conducted with individual and combined extract components incorporated into artificial diets on which Tribolium adults fed for six days. Feeding by T. brevicornis and T. castaneum on flour disks containing cuticular extracts of T. brevicornis pupae resulted in reduced consumption and weight loss relative to feeding on control flour disks. In both cases, feeding deterrence indices exceeded 80% suggesting that T. brevicornis cuticular hydrocarbons could function to deter cannibalism and predation of pupae by larvae and adult beetles. Sixteen different cuticular hydrocarbons were identified in T. brevicornis pupal extracts. Eight of the commercially available linear alkanes were tested individually in feeding trials with eight Tribolium species. One compound (C28) significantly reduced the amount of food consumed by three species compared to control disks, whereas the compounds C25, C26, and C27 elicited increased feeding in some species. Four other compounds had no effect on consumption for any species. When four hydrocarbon mixtures were tested for synergistic deterrence on T. brevicornis and T. castaneum, none significantly influenced consumption. Our results indicate that the cuticular chemistry of T. brevicornis pupae could serve to deter predation by conspecific and congeneric beetles.
Assuntos
Canibalismo , Hidrocarbonetos/química , Comportamento Predatório , Tribolium/química , Animais , Feminino , Hidrocarbonetos/farmacologia , Masculino , Comportamento Predatório/efeitos dos fármacos , Pupa/química , Pupa/efeitos dos fármacos , Tribolium/efeitos dos fármacosRESUMO
The aphid Aphis gossypii Glover (Homoptera: Aphididae) is known as worldwide important pest. It causes serious injuries to crops, but is also able to feed on various cultivated and wild plants. Some programs are developped to control this aphid. In many studies, hoverfy Episyrphus balteatus De Geer (Diptera: Syrphidae) is used as an efficient and aphid specific predator in biological control systems. In this work, we study cotton aphid choice plant discrimination, the effect of host plants on its developpment (fecondity and mortality), and the E. balteatus foraging and oviposition behavior according aphid host plants. Biologic parameters were observed using various plant combinations. E. balteatus behavioral observations were conducted using the Noldus Observer v.5.0. A. gossypii makes discriminant choice of his hosts, with better preference on Cucurbita pepo L. (Cucurbitaceae) and Capsicum annuum L. (Solanaceae). Dacryodes edulis (G. Don) H.J. Lam. (Burseraceae) and Vicia faba L. (Fabaceae) are the midle choice while Hibiscus sabdariffa L. (Malvaceae) is less chosen. A. gossypii develops easily on C. pepo, and weakly on Canuum, H. sabdariffa, and V. faba. D. edulis induces very high mortality to A. gossypii. According to E. balteatus behavior, A. gossypii is more attractive when infesting C. pepo and V. faba, less on, H. sabdariffa. While non-infested, V. faba appeared to be more attractive for the hoverfly, but without significant differnce compared to other plants. Our results highlited the importance of interactions between aphids and their host plants and the prey-host influence on a specific aphid predatory hoverfy. Other ways like melon aphid and its host plant volatils emission, in correlation with this work, are to investigated to complete this study and perform melon aphid biological control.
Assuntos
Afídeos/fisiologia , Dípteros/fisiologia , Herbivoria , Plantas , Animais , Afídeos/crescimento & desenvolvimento , Dípteros/crescimento & desenvolvimento , Oviposição , Controle Biológico de VetoresRESUMO
The Mi-1.2 gene in tomato confers resistance against certain clones of the potato aphid (Macrosiphum euphorbiae). This study used 2D-DIGE coupled with protein identification by MALDI-TOF-MS to compare the proteome patterns of avirulent and semivirulent potato aphids and their bacterial endosymbionts on resistant (Mi-1.2+) and susceptible (Mi-1.2-) tomato lines. Avirulent aphids had low survival on resistant plants, whereas the semivirulent clone could colonize these plants. Eighty-two protein spots showed significant quantitative differences among the four treatment groups, and of these, 48 could be assigned putative identities. Numerous structural proteins and enzymes associated with primary metabolism were more abundant in the semivirulent than in the avirulent aphid clone. Several proteins were also up-regulated in semivirulent aphids when they were transferred from susceptible to resistant plants. Nearly 25% of the differentially regulated proteins originated from aphid endosymbionts and not the aphid itself. Six were assigned to the primary endosymbiont Buchnera aphidicola, and 5 appeared to be derived from a Rickettsia-like secondary symbiont. These results indicate that symbiont expression patterns differ between aphid clones with differing levels of virulence, and are influenced by the aphids' host plant. Potentially, symbionts may contribute to differential adaptation of aphids to host plant resistance.
Assuntos
Afídeos/fisiologia , Proteômica , Solanum lycopersicum/parasitologia , Simbiose/fisiologia , Animais , Afídeos/química , Afídeos/microbiologia , Proteínas de Bactérias/isolamento & purificação , Proteínas do Citoesqueleto/análise , Proteínas do Citoesqueleto/isolamento & purificação , Eletroforese em Gel Bidimensional , Proteínas de Insetos/isolamento & purificação , Dados de Sequência Molecular , Proteínas de Plantas/isolamento & purificação , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/parasitologia , Reação em Cadeia da Polimerase , Rickettsia/química , Rickettsia/fisiologiaRESUMO
PCBs are persistent organic pollutants largely distributed in the biosphere. Although their effects on vertebrates are well described, little is known about their action on freshwater invertebrate's metabolism. Gammarus pulex (Linné) was selected as an indicator model to develop a proteomic approach in order to characterize the effects of PCBs on the protein profile of this freshwater crustacean. Sublethal coplanar PCBs exposition and related 2D gel were performed. More than 560 spots were detected and a total of 21 proteins exhibiting significant expression differences in PCB exposed to G. pulex were identified by mass spectrometry. Database searches were conducted to relate the results to well-known metabolic pathways (pentose phosphate, cytoskeleton, energy, etc.). In particular, glyceraldehyde 3-phosphate dehydrogenase and arginine kinase were found to be sensitive to the PCB exposition of G. pulex. The aim of the present study was to assess the biochemical responses and the metabolic changes in G. pulex following intoxication to coplanar PCB congeners CB77 and CB169 by a proteomic approach. This approach allowed us, by the identification of key proteins, to highlight important biochemical mechanisms disturbed by the presence of these contaminants in G. pulex.
Assuntos
Anfípodes/efeitos dos fármacos , Ecossistema , Bifenilos Policlorados/metabolismo , Proteômica/métodos , Poluentes Químicos da Água/metabolismo , Anfípodes/química , Anfípodes/metabolismo , Animais , Arginina Quinase/metabolismo , Biomarcadores/metabolismo , Cromatografia Gasosa , Bases de Dados de Proteínas , Eletroforese em Gel de Poliacrilamida , Água Doce/química , Glutamatos/metabolismo , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Glicólise/fisiologia , Longevidade/efeitos dos fármacos , Fosfopiruvato Hidratase/metabolismo , Bifenilos Policlorados/toxicidade , Análise Serial de Proteínas , Poluentes Químicos da Água/toxicidadeRESUMO
BACKGROUND: Aphid adaptation to harsh winter conditions is illustrated by an alternation of their reproductive mode. Aphids detect photoperiod shortening by sensing the length of the night and switch from viviparous parthenogenesis in spring and summer, to oviparous sexual reproduction in autumn. The photoperiodic signal is transduced from the head to the reproductive tract to change the fate of the future oocytes from mitotic diploid embryogenesis to haploid formation of gametes. This process takes place in three consecutive generations due to viviparous parthenogenesis. To understand the molecular basis of the switch in the reproductive mode, transcriptomic and proteomic approaches were used to detect significantly regulated transcripts and polypeptides in the heads of the pea aphid Acyrthosiphon pisum. RESULTS: The transcriptomic profiles of the heads of the first generation were slightly affected by photoperiod shortening. This suggests that trans-generation signalling between the grand-mothers and the viviparous embryos they contain is not essential. By analogy, many of the genes and some of the proteins regulated in the heads of the second generation are implicated in visual functions, photoreception and cuticle structure. The modification of the cuticle could be accompanied by a down-regulation of the N-beta-alanyldopamine pathway and desclerotization. In Drosophila, modification of the insulin pathway could cause a decrease of juvenile hormones in short-day reared aphids. CONCLUSION: This work led to the construction of hypotheses for photoperiodic regulation of the switch of the reproductive mode in aphids.
Assuntos
Afídeos/genética , Perfilação da Expressão Gênica , Fotoperíodo , Proteoma/metabolismo , Estações do Ano , Adaptação Fisiológica/genética , Animais , Afídeos/metabolismo , Afídeos/fisiologia , Dopamina/análogos & derivados , Dopamina/metabolismo , Regulação para Baixo , Feminino , Genes de Insetos , Cabeça , Análise de Sequência com Séries de Oligonucleotídeos , Partenogênese/genéticaRESUMO
Beekeepers suspected maize, Zea mays L., treated with imidacloprid to result in substantial loss of honey bee (Hymenoptera: Apidae) colonies in Belgium. The objective of this study was to investigate the potential impact of maize grown from imidacloprid-treated seeds on honey bee mortality. A survey of 16 apiaries was carried out, and all maize fields treated or not with imidacloprid were located within a radius of 3,000 m around the observed apiaries. Samples of honey, beeswax, and bees were collected in three colonies per apiary and analyzed for pesticide contain by liquid chromatography-tandem mass spectrometry and gas chromatography-tandem mass spectrometry. We first found a significant correlation between the number of colonies per apiary and the mortality rates in an apiary. In addition, this mortality rate was inversely correlated with the surface of maize fields treated and not with imidacloprid, suggesting that this pesticide do not interact with bees' fitness. Moreover, a very large number of our samples contained acaricides either prohibited or ineffective against Varroa destructor (Anderson & Trueman) (Acari: Varroidae), suggesting that the treatment methods used by the beekeepers to be inadequate for mite control. Our results support the hypothesis that imidacloprid seed-treated maize has no negative impact on honey bees.
Assuntos
Abelhas/efeitos dos fármacos , Imidazóis/farmacologia , Inseticidas/farmacologia , Nitrocompostos/farmacologia , Sementes , Zea mays , Animais , Mel/análise , Imidazóis/química , Inseticidas/química , Neonicotinoides , Nitrocompostos/química , Resíduos de Praguicidas/química , Ceras/químicaRESUMO
Forensic entomology uses pig carcasses to surrogate human decomposition and to investigate the entomofaunal colonization. Insects communicate with their environment through the use of chemical mediators, which in the case of necrophagous insects, may consist in the cadaveric volatile organic compounds (VOCs) released by the corpse under decomposition. Previous studies have focused on cadaveric VOCs released from human corpses. Nevertheless, studies on human corpses are restricted for many reasons, including ethics. Forensic entomologists use pig as animal model but very few information are available about the decompositional VOCs released by a decaying pig carcass. We here tested a passive sampling technique, the Radiello diffusive sampler, to monitor the cadaveric VOCs released by decomposing pig carcasses in three biotopes (crop field, forest, urban site). A total of 104 chemical compounds, exclusively produced by the decompositional process, were identified by thermal desorption interfaced with gas chromatography and mass spectrometry (TDS-GC-MS). Ninety, 85 and 57 cadaveric VOCs were identified on pig carcasses laying on the agricultural site, the forest biotope and in the urban site, respectively. The main cadaveric VOCs are acids, cyclic hydrocarbons, oxygenated compounds, sulfur and nitrogen compounds. A better knowledge of the smell of death and their volatile constituents may have many applications in forensic sciences.
Assuntos
Monitoramento Ambiental , Mudanças Depois da Morte , Volatilização , Ácidos/análise , Álcoois/análise , Aldeídos/análise , Animais , Meio Ambiente , Ésteres/análise , Patologia Legal , Cromatografia Gasosa-Espectrometria de Massas , Umidade , Hidrocarbonetos Acíclicos/análise , Hidrocarbonetos Cíclicos/análise , Cetonas/análise , Compostos de Nitrogênio/análise , Análise de Componente Principal , Compostos de Enxofre/análise , Suínos , Temperatura , ÁrvoresRESUMO
After the emergence of bluetongue (BT) in Belgium in 2006, two types of entomological surveys were initiated, the one to identify the local vector species, and the other to study their population dynamics. In the vector study, Culicoides were captured near farms with recently infected cattle or sheep; in the population study Culicoides were captured in two meadows situated in the BT-affected region. A total of 130 pools of parous, non-blood engorged female midges (with a mean of 7.5 midges per pool) were analysed with real-time reverse transcription PCR (RT-qPCR) targeting bluetongue virus (BTV) segment 5. To ensure the RNA integrity of the samples, all pools were also tested in a second RT-qPCR targeting Culicoides 18S rRNA, which served as an internal control. Seventeen pools with negative results for both 18S and BTV were excluded, most of which originated from the population survey. In the vector survey near outbreak sites, female midges of the obsoletus complex, including C. obsoletus, C. scoticus, C. dewulfi and C. chiopterus, dominated the black-light trap collections with 19 of 89 pools being BTV-positive. Moreover, all the collections from the vector survey included at least one positive pool of the obsoletus complex compared with only 20% collections (C. obsoletus/C. scoticus) in the population survey. The current study also revealed the presence of BTV RNA in one of five pools of C. pulicaris females captured near recent BT outbreaks, suggesting that this species might have played a role in transmission. Finally, the use of RT-qPCR for the recognition of new potential BTV vector species and the impact of an appropriate monitoring method and internal control are discussed.