Bluetongue Virus Serotype 3 and Schmallenberg Virus in Culicoides Biting Midges, Western Germany, 2023.

In October 2023, bluetongue virus serotype 3 (BTV-3) emerged in Germany, where Schmallenberg virus is enzootic. We detected BTV-3 in 1 pool of Culicoides biting midges collected at the time ruminant infections were reported. Schmallenberg virus was found in many vector pools. Vector trapping and analysis could elucidate viral spread.

Characterization of a natural 'dead-end' variant of Schmallenberg virus.

Schmallenberg virus (SBV) belongs to the Simbu serogroup within the family Peribunyaviridae, genus Orthobunyavirus and is transmitted by Culicoides biting midges. Infection of naïve ruminants in a critical phase of gestation may lead to severe congenital malformations. Sequence analysis from viremic animals revealed a very high genome stability. In contrast, sequence variations are frequently described for SBV from malformed fetuses. In addition to S segment mutations, especially within the M segment encoding the major immunogen Gc, point mutations or genomic deletions are also observed. Analysis of the SBV_D281/12 isolate from a malformed fetus revealed multiple point mutations in all three genome segments. It also has a large genomic deletion in the antigenic domain encoded by the M segment compared to the original SBV reference strain 'BH80/11' isolated from viremic blood in 2011. Interestingly, SBV_D281/12 showed a marked replication deficiency in vitro in Culicoides sonorensis cells (KC cells), but not in standard baby hamster kidney cells (BHK-21). We therefore generated a set of chimeric viruses of rSBV_D281/12 and wild-type rSBV_BH80/11 by reverse genetics, which were characterized in both KC and BHK-21 cells. It could be shown that the S segment of SBV_D281/12 is responsible for the replication deficit and that it acts independently from the large deletion within Gc. In addition, a single point mutation at position 111 (S to N) of the nucleoprotein was identified as the critical mutation. Our results suggest that virus variants found in malformed fetuses and carrying characteristic genomic mutations may have a clear 'loss of fitness' for their insect hosts in vitro. It can also be concluded that such mutations lead to virus variants that are no longer part of the natural transmission cycle between mammalian and insect hosts. Interestingly, analysis of a series of SBV sequences confirmed the S111N mutation exclusively in samples of malformed fetuses and not in blood from viremic animals. The characterization of these changes will allow the definition of protein functions that are critical for only one group of hosts.

Nestling sex and behaviour determine the host preference of insect vectors in avian nests.

Sexual differences in pathogen prevalence in wildlife often arise from varying susceptibility influenced by factors such as sex hormones and exposure to pathogens. In the case of vector-borne pathogens, host selection by insect vectors determines the exposure of hosts to infections, largely affecting the transmission of these infectious diseases. We identify the blood-feeding patterns of insect vectors in Blue Tit (Cyanistes caeruleus) nestlings in a 3-year study. Blood from both nestlings and insect vectors (Culicoides spp. and Simuliidae) captured inside nest-boxes were used to molecularly determine the sex of the host. We then compared the sex-ratios of the nestlings that had been bitten and those of the complete brood in each nest. We found that males were bitten more frequently than females in 2021, when males weighed less in comparison to other years. Additionally, we molecularly identified bitten nestlings individually by genotyping the DNA of blood obtained from both, the vector's abdomen and nestlings of each brood in 2022. Nestlings more frequently bitten by vectors were males, weighed less and were closest to the nest entrance. To our knowledge this is the first study identifying the nestling selection by insect vectors in bird nests under natural conditions. These results contribute to understanding the mechanisms of host selection by insect vectors, shedding light on pathogen transmission and offering insights into the observed sex-biased infections in wildlife populations.

Isolation and whole-genome sequence analysis of Balagodu virus in Japan.

Whole-genome sequencing of a virus isolated from Culicoides biting midges in southern Japan in 2020 revealed that it is a strain of Balagodu virus (BLGV; genus Orthobunyavirus; family Peribunyaviridae; order Bunyavirales). A solitary instance of BLGV isolation occurred in India in 1963. All assembled segments comprise complete protein-coding sequences that are similar to those of other orthobunyaviruses. The consensus 3'- and 5'-terminal sequences of orthobunyaviruses' genomic RNAs are also conserved in the Japanese BLGV strain. Here, we update the geographic distribution of BLGV and provide its complete sequence, contributing to the clarification of orthobunyavirus phylogeny.

Climate correlates of bluetongue incidence in southern Portugal.

Model forecasts of the spatiotemporal occurrence dynamics of diseases are necessary and can help understand and thus manage future disease outbreaks. In our study, we used ecological niche modelling to assess the impact of climate on the vector suitability for bluetongue disease, a disease affecting livestock production with important economic consequences. Specifically, we investigated the relationship between the occurrence of bluetongue outbreaks and the environmental suitability of each of the four vector species studied. We found that the main vector for bluetongue disease, Culicoides imicola, a typically tropical and subtropical species, was a strong predictor for disease outbreak occurrence in a region of southern Portugal from 2004 to 2021. The results highlight the importance of understanding the climatic factors that might influence vector presence to help manage infectious disease impacts. When diseases impact economically relevant species, the impacts go beyond mortality and have important economic consequences.

Ecological niche modelling of Culicoides imicola and future range shifts under climate change scenarios in Italy.

Culicoides imicola is the main vector of viral diseases of livestock in Europe such as bluetongue (BT), African horse sickness and epizootic haemorrhagic disease. Climatic factors are the main drivers of C. imicola occurrence and its distribution might be subject to rapid shifts due to climate change. Entomological data, collected during BT surveillance, and climatic/environmental variables were used to analyse ecological niche and to model C. imicola distribution and possible future range shifts in Italy. An ensemble technique was used to weigh the performance of machine learning, linear and profile methods. Updated future climate projections from the latest phase of the Climate Model Intercomparison Project were used to generate future distributions for the next three 20-year periods, according to combinations of general circulation models and shared socioeconomic pathways and considering different climate change scenarios. Results indicated the minimum temperature of the coldest month (BIO 6) and precipitation of the driest-warmest months (BIO 14) as the main limiting climatic factors. Indeed, BIO 6 and BIO 14 reported the two highest values of variable importance, respectively, 9.16% (confidence interval [CI] = 7.99%-10.32%), and 2.01% (CI = 1.57%-2.44%). Under the worst-case scenario of climate change, C. imicola range is expected to expand northward and shift away from the coasts of central Italy, while in some areas of southern Italy, environmental suitability will decrease. Our results provide predictions of C. imicola distribution according to the most up-to-date future climate projections and should be of great use to surveillance management at regional and national scales.

Culicoides biting midges feeding behaviour as a key for understanding avian Haemoproteus transmission in Lithuania.

Investigations of host feeding behaviour in haematophagous insects are critical to assess transmission routes of vector-borne diseases. Understanding if a certain species has ornithophilic or mammalophilic feeding behaviour can facilitate future studies focused on pathogens transmission to and from certain host species. Culicoides Latreille (Diptera: Ceratopogonidae) are vectors of several pathogens, which include arboviruses, bacteria and parasites to a considerable diversity of vertebrate hosts. However, most of the studies focused on feeding habits target Culicoides species that could transmit the Bluetongue virus, consequently with a mammalophilic feeding behaviour, leaving aside the Culicoides species that are involved in the transmission of vector-borne parasites to birds, such as Haemoproteus Kruse (Haemosporida: Haemoproteidae). This study aimed to investigate the source of blood meals of wild-caught Culicoides using molecular-based methods and to correlate our findings with the reports of Haemoproteus parasites in Culicoides species. Engorged Culicoides females were collected using ultraviolet (UV)-light traps at seven different localities in Lithuania in 2021-2023. Biting midges were dissected, and the abdomens of engorged females were used for molecular investigation of the blood meal source. A polymerase chain reaction (PCR) protocol that amplifies a fragment of the Cytochrome B gene of vertebrates was used. Obtained sequences were compared to available information in GenBank database to confirm the source of the blood meal. In total, 258 engorged Culicoides females, representing nine different species, were analysed. The source of blood meal was identified in 29.1% of them with most of the insects having fed on birds (74.7%) and the remaining on mammals (25.3%). Culicoides segnis Campbell, Pelham-Clinton was the only species to feed exclusively on birds; Culicoides from the Obsoletus group, C. pallidicornis Kieffer and C. punctatus Latreille were found to feed exclusively on mammals; C. festivipennis Kieffer, C. kibunensis Tokunaga and C. pictipennis Staeger had an opportunistic feeding behaviour, with the first two preferably feeding on birds. Due to their feeding behaviour and the presence of Haemoproteus parasites reported in the literature, C. festivipennis, C. kibunensis, C. pictipennis, and C. segnis play an important role in the transmission of those avian vector-borne parasite in the wild. These Culicoides species were already confirmed as being able to support the development of several Haemoproteus species and lineages. Future studies focused on understanding the epidemiology of avian pathogens transmitted by Culicoides should target these species.

Evaluating Temperature Effects on Bluetongue Virus Serotype 10 and 17 Coinfection in Culicoides sonorensis.

Bluetongue virus (BTV) is a segmented, double-stranded RNA virus transmitted by Culicoides midges that infects ruminants. As global temperatures increase and geographical ranges of midges expand, there is increased potential for BTV outbreaks from incursions of novel serotypes into endemic regions. However, an understanding of the effect of temperature on reassortment is lacking. The objectives of this study were to compare how temperature affected Culicoides survival, virogenesis, and reassortment in Culicoides sonorensis coinfected with two BTV serotypes. Midges were fed blood meals containing BTV-10, BTV-17, or BTV serotype 10 and 17 and maintained at 20 °C, 25 °C, or 30 °C. Midge survival was assessed, and pools of midges were collected every other day to evaluate virogenesis of BTV via qRT-PCR. Additional pools of coinfected midges were collected for BTV plaque isolation. The genotypes of plaques were determined using next-generation sequencing. Warmer temperatures impacted traits related to vector competence in offsetting ways: BTV replicated faster in midges at warmer temperatures, but midges did not survive as long. Overall, plaques with BTV-17 genotype dominated, but BTV-10 was detected in some plaques, suggesting parental strain fitness may play a role in reassortment outcomes. Temperature adds an important dimension to host-pathogen interactions with implications for transmission and evolution.

Visualisation of Bluetongue Virus in the Salivary Apparatus of Culicoides Biting Midges Highlights the Accessory Glands as a Primary Arboviral Infection Site.

BACKGROUND: Arthropods transmit a wide range of pathogens of importance for the global health of humans, animals, and plants. One group of these arthropod vectors, Culicoides biting midges (Diptera: Ceratopogonidae), is the biological vector of several human and animal pathogens, including economically important livestock viruses like bluetongue virus (BTV). Like other arthropod-borne viruses (arboviruses), Culicoides-borne viruses must reach and replicate in the salivary apparatus, from where they can be transmitted to susceptible hosts through the saliva during subsequent blood feeding. Despite the importance of the salivary gland apparatus for pathogen transmission to susceptible animals from the bite of infected Culicoides, these structures have received relatively little attention, perhaps due to the small size and fragility of these vectors. RESULTS: In this study, we developed techniques to visualize the infection of the salivary glands and other soft tissues with BTV, in some of the smallest known arbovirus vectors, Culicoides biting midges, using three-dimensional immunofluorescence confocal microscopy. We showed BTV infection of specific structures of the salivary gland apparatus of female Culicoides vectors following oral virus uptake, related visualisation of viral infection in the salivary apparatus to high viral RNA copies in the body, and demonstrated for the first time, that the accessory glands are a primary site for BTV replication within the salivary apparatus. CONCLUSIONS: Our work has revealed a novel site of virus-vector interactions, and a novel role of the accessory glands of Culicoides in arbovirus amplification and transmission. Our approach would also be applicable to a wide range of arbovirus vector groups including sand flies (Diptera: Psychodidae), as well as provide a powerful tool to investigate arbovirus infection and dissemination, particularly where there are practical challenges in the visualization of small size and delicate tissues of arthropods.

The Acquisition and Retention of Lumpy Skin Disease Virus by Blood-Feeding Insects Is Influenced by the Source of Virus, the Insect Body Part, and the Time since Feeding.

Lumpy skin disease virus (LSDV) is a poxvirus that causes severe systemic disease in cattle and is spread by mechanical arthropod-borne transmission. This study quantified the acquisition and retention of LSDV by four species of Diptera (Stomoxys calcitrans, Aedes aegypti, Culex quinquefasciatus, and Culicoides nubeculosus) from cutaneous lesions, normal skin, and blood from a clinically affected animal. The acquisition and retention of LSDV by Ae. aegypti from an artificial membrane feeding system was also examined. Mathematical models of the data were generated to identify the parameters which influence insect acquisition and retention of LSDV. For all four insect species, the probability of acquiring LSDV was substantially greater when feeding on a lesion compared with feeding on normal skin or blood from a clinically affected animal. After feeding on a skin lesion LSDV was retained on the proboscis for a similar length of time (around 9 days) for all four species and for a shorter time in the rest of the body, ranging from 2.2 to 6.4 days. Acquisition and retention of LSDV by Ae. aegypti after feeding on an artificial membrane feeding system that contained a high titer of LSDV was comparable to feeding on a skin lesion on a clinically affected animal, supporting the use of this laboratory model as a replacement for some animal studies. This work reveals that the cutaneous lesions of LSD provide the high-titer source required for acquisition of the virus by insects, thereby enabling the mechanical vector-borne transmission. IMPORTANCE Lumpy skin disease virus (LSDV) is a high consequence pathogen of cattle that is rapidly expanding its geographical boundaries into new regions such as Europe and Asia. This expansion is promoted by the mechanical transmission of the virus via hematogenous arthropods. This study quantifies the acquisition and retention of LSDV by four species of blood-feeding insects and reveals that the cutaneous lesions of LSD provide the high titer virus source necessary for virus acquisition by the insects. An artificial membrane feeding system containing a high titer of LSDV was shown to be comparable to a skin lesion on a clinically affected animal when used as a virus source. This promotes the use of these laboratory-based systems as replacements for some animal studies. Overall, this work advances our understanding of the mechanical vector-borne transmission of LSDV and provides evidence to support the design of more effective disease control programmes.

Comparative viromes of Culicoides and mosquitoes reveal their consistency and diversity in viral profiles.

The genus Culicoides includes biting midges, some of which are vectors for viruses that cause diseases in humans and animals. Knowledge of the roles of Culicoides in viral ecology is inadequate. We collected ~300 000 samples of Culicoides and mosquitoes in 15 representative regions within Yunnan, China. Using mosquitoes as reference vectors, we designed a comparative virome strategy to study the viral composition, diversity, hosts and spatiotemporal distribution of Culicoides. A map of viromes in Culicoides and mosquitoes in Yunan province, China, was constructed. At the same locations, Culicoides and mosquitoes usually share a similar viral diversity. At least 10 important pathogenic viruses were detected from Culicoides. Many novel viruses were discovered, including 21 segmented viruses of Flaviviridae, 180 viruses of Monjiviricetes and 130 viruses of Bunyavirales. The findings demonstrate that Culicoides is an important part of viral ecology and should be studied and monitored for potentially emerging viruses.

An Agent-Based Model of Biting Midge Dynamics to Understand Bluetongue Outbreaks.

Bluetongue (BT) is a well-known vector-borne disease that infects ruminants such as sheep, cattle, and deer with high mortality rates. Recent outbreaks in Europe highlight the importance of understanding vector-host dynamics and potential courses of action to mitigate the damage that can be done by BT. We present an agent-based model, entitled 'MidgePy', that focuses on the movement of individual Culicoides spp. biting midges and their interactions with ruminants to understand their role as vectors in BT outbreaks, especially in regions that do not regularly experience outbreaks. The results of our sensitivity analysis suggest that midge survival rate has a significant impact on the probability of a BTV outbreak as well as its severity. Using midge flight activity as a proxy for temperature, we found that an increase in environmental temperature corresponded with an increased probability of outbreak after identifying parameter regions where outbreaks are more likely to occur. This suggests that future methods to control BT spread could combine large-scale vaccination programs with biting midge population control measures such as the use of pesticides. Spatial heterogeneity in the environment is also explored to give insight on optimal farm layouts to reduce the potential for BT outbreaks.

The use of path analysis to determine effects of environmental factors on the adult seasonality of Culicoides (Diptera: Ceratopogonidae) vector species in Spain.

Culicoides biting midges (Diptera: Ceratopogonidae) are the main vectors of livestock diseases such as bluetongue (BT) which mainly affect sheep and cattle. In Spain, bluetongue virus (BTV) is transmitted by several Culicoides taxa, including Culicoides imicola, Obsoletus complex, Culicoides newsteadi and Culicoides pulicaris that vary in seasonality and distribution, affecting the distribution and dynamics of BT outbreaks. Path analysis is useful for separating direct and indirect, biotic and abiotic determinants of species' population performance and is ideal for understanding the sensitivity of adult Culicoides dynamics to multiple environmental drivers. Start, end of season and length of overwintering of adult Culicoides were analysed across 329 sites in Spain sampled from 2005 to 2010 during the National Entomosurveillance Program for BTV with path analysis, to determine the direct and indirect effects of land use, climate and host factor variables. Culicoides taxa had species-specific responses to environmental variables. While the seasonality of adult C. imicola was strongly affected by topography, temperature, cover of agro-forestry and sclerophyllous vegetation, rainfall, livestock density, photoperiod in autumn and the abundance of Culicoides females, Obsoletus complex species seasonality was affected by land-use variables such as cover of natural grassland and broad-leaved forest. Culicoides female abundance was the most explanatory variable for the seasonality of C. newsteadi, while C. pulicaris showed that temperature during winter and the photoperiod in November had a strong effect on the start of the season and the length of overwinter period of this species. These results indicate that the seasonal vector-free period (SVFP) in Spain will vary between competent vector taxa and geographic locations, dependent on the different responses of each taxa to environmental conditions.

Effect of substrate salinity and pH on life history traits of the bluetongue virus vector Culicoides peregrinus.

Habitat selection of Culicoides spp. (Diptera: Ceratopogonidae) is influenced by the physicochemical factors such as temperature, pH, salinity, moisture, conductivity, organic and inorganic compounds of substrates. These factors determine the life history traits of the vectors. We studied the influence of substrate salinity (0-40 parts per thousand, ppt) and pH (pH 1-13) on oviposition, egg hatching, larval survivability, and adult emergence of Culicoides peregrinus Kieffer under laboratory conditions. Most eggs (80.74%) were laid in 0 ppt and 95% in pH 7 but lowered with increased salinity and pH levels. It was observed that the females did not lay eggs in 30 ppt to 40 ppt salinity; pH 1 and pH 13 but interestingly up to 95% of the eggs were retained within the abdomen. Little effect of salinity and pH on egg hatching was observed up to 5 ppt and 10 ppt except at the extreme values of 40 ppt and pH 1, pH 13. Pupation did not occur in rearing plates with high salinities, 30 ppt and 40 ppt, although the few eggs hatched when exposed to such salinity. In low salinity (0 to 2 ppt), occurrence of adult emergence was more and then decreased with increasing salinity. Maximum emergence was seen when the rearing media was alkaline. This study deals with the suitability of breeding substrate of C. peregrinus when exposed to salinity and pH ranges. Our study suggests the ambient salinity and pH ranges to be maintained during laboratory rearing of this vector species.

Vector communities under global change may exacerbate and redistribute infectious disease risk.

Vector-borne parasites may be transmitted by multiple vector species, resulting in an increased risk of transmission, potentially at larger spatial scales compared to any single vector species. Additionally, the different abilities of patchily distributed vector species to acquire and transmit parasites will lead to varying degrees of transmission risk. Investigation of how vector community composition and parasite transmission change over space due to variation in environmental conditions may help to explain current patterns in diseases but also informs our understanding of how patterns will change under climate and land-use change. We developed a novel statistical approach using a multi-year, spatially extensive case study involving a vector-borne virus affecting white-tailed deer transmitted by Culicoides midges. We characterized the structure of vector communities, established the ecological gradient controlling change in structure, and related the ecology and structure to the amount of disease reporting observed in host populations. We found that vector species largely occur and replace each other as groups, rather than individual species. Moreover, community structure is primarily controlled by temperature ranges, with certain communities being consistently associated with high levels of disease reporting. These communities are essentially composed of species previously undocumented as potential vectors, whereas communities containing putative vector species were largely associated with low levels, or even absence, of disease reporting. We contend that the application of metacommunity ecology to vector-borne infectious disease ecology can greatly aid the identification of transmission hotspots and an understanding of the ecological drivers of parasite transmission risk both now and in the future.

Parasite load effects on sex ratio, size, survival and mating fitness of Heleidomermis magnapapula in Culicoides sonorensis.

Heleidomermis magnapapula parasitizes the blood-feeding midge Culicoides sonorensis. Most (84%) single mermithid infective second stage juveniles (J2) developed into adult females, while parasitism by multiple J2 yielded 97% male adults. Nematodes emerged from the midge larval host as adults and mated immediately; females were ovoviviparous. Host larvae were exposed to nematode J2 and examined intact microscopically to score initial parasite load. Midge hosts were reared individually. Premature midge death, nematode survival within the host, and emerging adult nematode sex ratio and size as a function of load and host size were all tracked. Higher nematode loads produced smaller adult nematode males. The higher loads also increased and accelerated premature host death. Emergence of > 7-9 adult nematode males was rare, but up to 19 tiny males emerged from a single host. Larger midges supported higher parasite loads and a larger total volume of emerged nematode biomass. Virgin adult nematode males then were paired with females of variable, known sizes (volume) and held to determine size effects on fertility (egg hatch), and male survival (longevity). Tested adult males ranged in size from 0.0025 - 0.0334 mm3 and females from 0.0121 - 0.1110 mm3. Logistic regression indicated female nematode fertility was positively influenced by male nematode size, while nematode load and female nematode size had no significant effect. While fertility was reduced statistically in smaller males, even some of the smallest male and female individuals could be fertile. Findings are related to field studies in this system.

Species Identity, Life History, and Geographic Distance Influence Gut Bacterial Communities in Lab-Reared and European Field-Collected Culicoides Biting midges.

Bacteria are part of the insect gut system and influence many physiological traits of their host. Gut bacteria may even reduce or block the transmission of arboviruses in several species of arthropod vectors. Culicoides biting midges are important arboviral vectors of several livestock and wildlife diseases, yet limited information is available on their gut bacterial communities. Addressing this gap will help inform how these communities can be manipulated and ultimately used as novel tools to control pathogens. To assess how bacterial communities change during the life stages of lab-reared C. nubeculosus and C. sonorensis, endosymbiotic bacteria were identified using Illumina sequencing of 16S rRNA and taxonomically characterised. Analyses were conducted to determine how gut bacterial communities in adults are influenced by species identity and geographic distance among biting midge populations. Communities of the two lab-reared Culicoides species significantly changed after pupation and with maturation into 6-day-old adults. Pseudomonas, Burkholderiaceae and Leucobacter bacteria were part of a core community that was trans-stadially transmitted and found throughout their life cycle. Among field-collected biting midges, the bacterial communities were unique for almost each species. Cardinium, Rickettsia and Wolbachia were some of the most abundant bacteria in midges collected from wetlands. Only Pseudomonas was present in high relative abundance in all field-collected species. In this study, species identity, as well as geographic distance, influenced the gut bacterial communities and may partly explain known inter- and intra-species variability in vector competence. Additionally, stably associated bacterial species could be candidates for paratransgenic strategies to control vector-borne pathogens.

Cattle-Derived Unsaturated Aldehydes Repel Biting Midges and Mosquitoes.

Host-derived repellents offer a novel way to reduce disease vector-host interactions, particularly for vectors and nuisance pests where commercial repellents are not available, e.g., Culicoides biting midges. By revising the criteria previously used to identify bioactive volatile organic compounds for Culicoides nubeculosus, we identify (E)-2-hexenal, (E)-2-heptenal, (E)-2-octenal, and (E)-2-nonenal to be differentially present in the headspace odour of cattle hair and to elicit antennal responses in this research model species. A blend of these unsaturated aldehydes elicited an aversive response in C. nubeculosus, as well as a repellent response in three disease vector mosquito species, Aedes aegypti, Culex quinquefasciatus and Anopheles coluzzii, a response that was stronger than that to the commercially available repellents tested (DEET, IR3535, PMD, icaridin, and d-allethrin). Culicoides nubeculosus was behaviourally indifferent to these commercially available repellents tested, except d-allethrin to which it was attracted. The identification of a host-derived repellent odour blend, which reduces the interaction between biting midges extends the array of tools to be used in integrated vector management of these and other disease vectors.

Comparison of the efficiency of the Onderstepoort- and Centres for Disease Control ultraviolet light traps for the collection of livestock associated Culicoides species in South Africa.

Comparative monitoring of the abundance and distribution of Culicoides biting midges (Diptera: Ceratopogonidae), the biological vectors of the causative agents of several diseases of global veterinary importance, will be crucial in determining the risk of disease outbreak and spread. Ultraviolet (UV) suction traps have become the most frequent method used for the monitoring of Culicoides diversity and abundance. The current study compared the trapping efficiency of the two most used UV suction light traps, i.e., the Onderstepoort (OP)- and the Centres for Disease Control trap, for the collection of livestock associated Culicoides species in South Africa. The study confirmed the superiority of the OP trap and indicated a correlation in species composition and age grading results as determine with the two trap types. Substantial variations in the comparative trap efficiency, as found between areas and sites within an area, suggest that a universal conversion factor between the two trap types may not be advisable as it is unclear to what extent species composition and environmental factors may influence the conversion factor. Light traps, independent of trap model, can be considered acceptable for determining the serial comparison of population numbers for seasonal fluctuation and species abundance in distribution surveys.

Putative roles of mosquitoes (Culicidae) and biting midges (Culicoides spp.) as mechanical or biological vectors of lumpy skin disease virus.

The stable fly Stomoxys calcitrans (Diptera: Muscidae) is considered as the main mechanical vector of the lumpy skin disease virus (LSDV). In addition, the mosquito species Aedes aegypti (Diptera: Culicidae) was shown to transmit the virus from donor to receptor animals. Retention of the virus for several days was shown for two additional tropical mosquito species and the biting midge Culicoides nubeculosus (Diptera: Ceratopogonidae). In the present study, viral retention for 10- or 7-days post feeding on virus-spiked blood through a membrane was shown for field-collected Aedes japonicus and laboratory-reared Culex pipiens, two widely distributed mosquito species in temperate regions. Viral DNA could be detected from honey-coated Flinders Technology Associates (FTA) cards and shedded faeces for 1 or 4 days after an infectious blood meal was given to Ae. aegypti. Virus increase over time and virus dissemination was observed in laboratory-reared C. nubeculosus, but the virus could be isolated from field-collected biting midges only from the day of exposure to the blood meal. Thus, mosquitoes might serve as mechanical vectors of LSDV in case of interrupted feeding. A putative biological virus transmission by Culicoides biting midges, as suspected from field observations, deserves further investigations.