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Epizootic Shell Disease (ESD) has posed a great threat, both ecologically and economically, to the American lobster population of Long Island Sound since its emergence in the late 1990s. Because of the polymicrobial nature of carapace infections, causative agents for ESD remain unclear. In this study, we aimed to identify carapace microbiota associated with ESD and its potential impact on the microbiota of internal organs (green gland, hepatopancreas, intestine, and testis) using high-throughput 16S rRNA gene sequencing. We found that lobsters with ESD harbored specific carapace microbiota characterized by high abundance of Aquimarina, which was significantly different from healthy lobsters. PICRUSt analysis showed that metabolic pathways such as amino acid metabolism were enriched in the carapace microbiota of lobsters with ESD. Aquimarina, Halocynthiibacter, and Tenacibaculum were identified as core carapace bacteria associated with ESD. Particularly, Aquimarina and Halocynthiibacter were detected in the green gland, hepatopancreas, and testis of lobsters with ESD, but were absent from all internal organs tested in healthy lobsters. Hierarchical clustering analysis revealed that the carapace microbiota of lobsters with ESD was closely related to the green gland microbiota, whereas the carapace microbiota of healthy lobsters was more similar to the testis microbiota. Taken together, our findings suggest that ESD is associated with alterations in the structure and function of carapace microbiota, which may facilitate the invasion of bacteria into the green gland. [ABSTRACT FROM AUTHOR]

Bluetongue virus (BTV) is an arbovirus that is transmitted between domestic and wild ruminants by Culicoides spp. Its worldwide distribution depends on competent vectors and suitable environmental ecosystems that are becoming affected by climate change. Therefore, we evaluated whether climate change would influence the potential distribution and ecological niche of BTV and Culicoides insignis in Peru. Here, we analyzed BTV (n = 145) and C. insignis (n = 22) occurrence records under two shared socioeconomic pathway scenarios (SSP126 and SSP585) with five primary general circulation models (GCMs) using the kuenm R package v.1.1.9. Then, we obtained binary presence–absence maps and represented the risk of transmission of BTV and niche overlapping. The niche model approach showed that north and east Peru presented suitability in the current climate scenario and they would have a decreased risk of BTV, whilst its vector would be stable and expand with high agreement for the five GCMs. In addition, its niche overlap showed that the two niches almost overlap at present and would completely overlap with one another in future climate scenarios. These findings might be used to determine the areas of highest priority for entomological and virological investigations and surveillance in order to control and prevent bluetongue infections in Peru. [ABSTRACT FROM AUTHOR]

The distribution pattern of the fly Chrysomya megacephala in the city of Yaoundé in Cameroon is poorly known despite its medical relevance. Following three types of standing (urbanization level) (medium standing, spontaneous standing, and peri-urban standing), we conducted monthly catches of adult flies on three types of waste (organic, inorganic, and animal paddock). The occurrence, distribution pattern, and seasonal abundance associated with abiotic factors (temperature, relative humidity, and precipitation) of C. megacephala were studied. In total, 656 fly individuals were sampled across all sites, with the highest abundance observed in mixed organic waste in spontaneous standings. Seasonal fluctuations revealed two peaks of C. megacephala abundance, one during the long dry season and the other during the short rainy season. Spearman's correlation test showed that fly's abundance is not significantly linked to temperature, relative humidity and rainfall. The occurrences data revealed that C. megacephala in the urban area of Yaoundé, C. megacephala was constant, incidental or accidental species. Chrysomya megacephala was constant only in mixed organic matter of spontaneous standings. The Fisher, Neyman and Morisita indices show that the mode of distribution of C. megacephala was aggregative. Current vector control strategies should also target sarcophagus insects like C. megacephala to reduce its abundance and the burden of hydric diseases in Yaoundé neighborhoods. [ABSTRACT FROM AUTHOR]

Simple Summary: This study is to investigate the potential vectors of African horse sickness (AHS). Culicoides were collected near horse stables in Hua Hin district, Prachuab Khiri Khan province, Thailand, where horses were affected and died from AHS in 2020. Twelve Culicoides species were identified. The predominant Culicoides species in all farms was C. oxystoma followed by C. imicola. The Culicoides collected in this study fed on blood from horses, dogs, pigs, and humans for their blood meal. This study has identified the potential AHS vector Culicoides species and its zoonotic potential in this area for the first time. African horse sickness (AHS) was reported as an outbreak in Thailand in 2020. Hematophagous insects from the genus Culicoides are the suspected vector responsible for AHS transmission. Horses in Hua Hin district, Prachuab Khiri Khan province, Thailand, were affected and died from AHS in 2020. However, the potential Culicoides species and its host preference blood meal in the affected areas are unknown. To investigate the potential vectors of AHS, Culicoides were collected using ultraviolet light traps placed near horse stables. Six horse farms, including five farms with AHS history and one farm without AHS history, were included in this study. Morphological and molecular identification of the Culicoides species was performed. Polymerase chain reaction (PCR) targeting the cytochrome b oxidase I (COXI) gene for confirmation of the Culicoides species, identification of the prepronociceptin (PNOC) gene for host preference blood meal, and bidirectional sequencing were conducted. Consequently, 1008 female Culicoides were collected, consisting of 708 and 300 samples captured at positions A and B at a distance of <2 and >5 m from the horse, respectively. Twelve Culicoides species identified by morphology were noted, including C. oxystoma (71.92%), C. imicola (20.44%), C. actoni (2.28%), C. flavipunctatus (1.98%), C. asiana (0.99%), C. peregrinus (0.60%), C. huffi (0.60%), C. brevitarsis (0.40%), C. innoxius (0.30%), C. histrio (0.30%), C. minimus (0.10%), and C. geminus (0.10%). The PCR detection of the Culicoides COXI gene confirmed Culicoides species in 23 DNA samples. PCR targeting the PNOC gene revealed that the Culicoides collected in this study fed on Equus caballus (86.25%), Canis lupus familiaris (6.25%), Sus scrofa (3.75%), and Homo sapiens (3.75%) for their blood meal. Human blood was identified from two samples of C. oxystoma and a sample of C. imicola. Three dominant species including C. oxystoma, C. imicola, and C. actoni that were reported in the Hua Hin area prefer to feed on horse blood. Moreover, C. oxystoma, C. imicola, and C. bravatarsis also feed on canine blood. This study revealed the species of Culicoides in Hua Hin district, Thailand, after the AHS outbreak. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

A newly described species, Dasyhelea mesophylla Dominiak and Borkent, from Costa Rica is the first species in this large and diverse family known to be a leaf-miner. The species is described as third and fourth instar larvae, pupa, and male and female adults. The biology of larvae mining the floating leaves of Salvinia minima Baker is also described. The pupa has a distinctive, sharply pointed respiratory organ, shared with some other species of Dasyhelea, which pierces the surface of the leaf to breathe. The species belongs in the grisea group and its close relatives within that group are noted. [ABSTRACT FROM AUTHOR]

Problems related to the identity of Simuliidae species are impediments to effective disease control in Amazonia. Some of these species, such as Simulium oyapockense Floch & Abonnenc, 1946 (Diptera: Simuliidae), are vectors of the organisms that cause onchocerciasis and mansonellosis diseases. This blackfly species has a wide distribution in South America, and it is suspected of being a complex of cryptic species. The aim of this study is to characterize the nominal species S. oyapockense using partial COI gene sequences. Seven populations of S. oyapockense (morphologically identified) were analysed, including one from its type‐locality. The other six populations were collected in Brazil and in Argentina. A taxon collected in Amazonas state, Brazil, with adults similar to S. oyapockense but with distinct pupae, was also included in the analysis (Simulium 'S'). The nominal species S. oyapockense is circumscribed, and its geographical distribution is restricted to areas north of the Amazon River. Populations of S. oyapockense s.l. collected south of the Amazon River comprise a species complex that needs to be evaluated using integrative taxonomy. Simulium 'S' represents a species with unique morphological and molecular characteristics. Distinguishing cryptic species is a prerequisite for reducing the taxonomic impediment, especially in medically important taxa. [ABSTRACT FROM AUTHOR]

Tibet orbivirus (TIBOV) was identified as a novel orbivirus in 2014. Antibodies against TIBOV were detected in cattle, Asian buffalo, and goats, while all the sequenced TIBOV strains were isolated from mosquitos and Culicoides. The known TIBOV strains have been classified into four putative serotypes. In this study, two TIBOV strains isolated from Culicoides spp. in Shizong County of Yunnan Province, China, were fully sequenced. The phylogenetic analysis of outer capsid protein 2 (VP2) indicated that these two viral strains belong to two novel putative serotypes of TIBOV. The updated putative serotypes may help in an investigation of the distribution and virulence of TIBOV. [ABSTRACT FROM AUTHOR]

The prevalence of autochthonous leishmaniasis in Thailand is increasing but the natural vectors that are responsible for transmission remain unknown. Experimental in vivo infections in Culicoides spp. with Leishmania (Mundinia) martiniquensis and Leishmania (Mundinia) orientalis, the major causative pathogens in Thailand, have demonstrated that biting midges can act as competent vectors. Therefore, the isolation and detection of Leishmania and other trypanosomatids were performed in biting midges collected at a field site in an endemic area of leishmaniasis in Tha Ruea and a mixed farm of chickens, goats, and cattle in Khuan Phang, Nakhon Si Thammarat province, southern Thailand. Results showed that Culicoides peregrinus was the abundant species (>84%) found in both locations and only cow blood DNA was detected in engorged females. Microscopic examination revealed various forms of Leishmania promastigotes in the foregut of several C. peregrinus in the absence of bloodmeal remnants, indicating established infections. Molecular identification using ITS1 and 3’UTR HSP70 type I markers showed that the Leishmania parasites found in the midges were L. martiniquensis. The infection rate of L. martiniquensis in the collected flies was 2% in Tha Ruea and 6% in Khuan Phang, but no L. orientalis DNA or parasites were found. Additionally, organisms from two different clades of Crithidia, both possibly new species, were identified using SSU rRNA and gGAPDH genes. Choanomastigotes and promastigotes of both Crithidia spp. were observed in the hindgut of the dissected C. peregrinus. Interestingly, midges infected with both L. martiniquensis and Crithidia were found. Moreover, four strains of Crithidia from one of the clades were successfully isolated into culture. These parasites could grow at 37°C in the culture and infect BALB/c mice macrophages but no multiplication was observed, suggesting they are thermotolerant monoxenous trypanosomatids similar to Cr. thermophila. These findings provide the first evidence of natural infection of L. martiniquensis in C. peregrinus supporting it as a potential vector of L. martiniquensis.

Haemoproteus parasites are the most diverse among Haemosporida. However, their natural vectors (Culicoides) are still poorly investigated and were identified for only a few parasite species and lineages. The application of an integrative approach (insect dissection, microscopic analysis, and molecular-based methods) is necessary in these studies, which have been carried out by a few research groups, mainly in Europe. The aim of this study was (i) to determine the Culicoides species that are naturally infected by Haemoproteus parasites, and which can support its complete sporogonic development, and (ii) to investigate the prevalence of Culicoides species and Haemoproteus parasite lineages in different study sites. In total, 1953 parous Culicoides females, from 11 species, were collected in four different localities in Lithuania and were dissected and analyzed using an integrative approach. The most abundant was C. pictipennis (30.3%). Parasite DNA was found in 7.9% of all investigated Culicoides, of which ~30% had sporozoites in their salivary glands, confirming their vector competence for these parasites. The Botanical Garden presented the highest number of Culicoides parous females, Culicoides species, and parasite lineages, as well as the highest positivity for sporozoites. Culicoides reconditus was confirmed as a natural vector of Haemoproteus parasites, sporozoites of six Haemoproteus lineages were reported for the first time, and 12 new interactions between Haemoproteus parasite lineages and Culicoides species were identified. Haemoproteus parasites seem to be transmitted by a high number of Culicoides species, with C. kibunensis, C. pictipennis, and C. segnis being the most important vectors.

Epizootic hemorrhagic disease (EHD) is a non-contagious arthropod-transmitted viral disease and a World Organization for Animal Health (WOAH)-listed disease of domestic and wild ruminants since 2008. EHDV is transmitted among susceptible animals by a few species of midges of genus Culicoides. During the fall of 2021, a large outbreak caused by the epizootic hemorrhagic disease virus (EHDV), identified as serotype 8, was reported in Tunisian dairy and beef farms with Bluetongue virus (BTV)-like clinical signs. The disease was detected later in the south of Italy, in Spain, in Portugal and, more recently, in France, where it caused severe infections in cattle. This was the first evidence of EHDV-8 circulation outside Australia since 1982. In this study, we analyzed the epidemiological situation of the 2021–2022 EHDV outbreaks reported in Tunisia, providing a detailed description of the spatiotemporal evolution of the disease. We attempted to identify the eco-climatic factors associated with infected areas using generalized linear models (GLMs). Our results demonstrated that environmental factors mostly associated with the presence of C. imicola, such as digital elevation model (DEM), slope, normalized difference vegetation index (NDVI), and night-time land surface temperature (NLST)) were by far the most explanatory variables for EHD repartition cases in Tunisia that may have consequences in neighboring countries, both in Africa and Europe through the spread of infected vectors. The risk maps elaborated could be useful for disease control and prevention strategies.

Bluetongue virus (BTV) is a segmented, double-stranded RNA orbivirus listed by the World Organization for Animal Health and transmitted by Culicoides biting midges. Segmented viruses can reassort, which facilitates rapid and important genotypic changes. Our study evaluated reassortment in Culicoides sonorensis midges coinfected with different ratios of BTV-10 and BTV-17. Midges were fed blood containing BTV-10, BTV-17, or a combination of both serotypes at 90:10, 75:25, 50:50, 25:75, or 10:90 ratios. Midges were collected every other day and tested for infection using pan BTV and cox1 (housekeeping gene) qRT-PCR. A curve was fit to the ∆Ct values (pan BTV Ct—cox1 Ct) for each experimental group. On day 10, the midges were processed for BTV plaque isolation. Genotypes of the plaques were determined by next-generation sequencing. Pairwise comparison of ∆Ct curves demonstrated no differences in viral RNA levels between coinfected treatment groups. Plaque genotyping indicated that most plaques fully aligned with one of the parental strains; however, reassortants were detected, and in the 75:25 pool, most plaques were reassortant. Reassortant prevalence may be maximized upon the occurrence of reassortant genotypes that can outcompete the parental genotypes. BTV reassortment and resulting biological consequences are important elements to understanding orbivirus emergence and evolution.

Avian haemosporidians (Apicomplexa, Haemosporida) are widespread blood protists, often causing severe haemosporidiosis, pathology, or even mortality in their hosts. Migrant birds regularly bring various haemosporidian parasites from wintering grounds to European breeding areas. Some haemosporidian parasites are prevalent in breeding sites and complete their life cycles in temperate climate zones and can be transmitted, but others do not. The factors altering the spread of these haemosporidians are not fully understood. Culicoides biting midges (Diptera: Ceratopogonidae) play an important role in the transmission of worldwide distributed avian haemosporidian parasites belonging to the genus Haemoproteus, but this information is particularly scarce and insufficient. The key factors limiting the spread of these pathogens in temperate climate zones, which we suspect and aim to study, are the absence of susceptible vectors and the ecological isolation of birds from vectors during the breeding period when transmission occurs. The primary objective of this study was to evaluate how the habitats of biting midges and bird breeding sites influence parasite transmission while also seeking to expand our understanding of the natural vectors for these parasites. Biting midges were collected using UV traps on the Curonian Spit, Lithuania, in different habitats, such as woodland and reeds, from May to September. Parous Culicoides females were identified, dissected, and investigated for the presence of Haemoproteus parasites using both microscopy and PCR-based tools. Among the dissected 1135 parous Culicoides females, the sporozoites of Haemoproteus asymmetricus (genetic lineage hTUPHI01) have been detected for the first time in the salivary glands of Culicoides festivipennis. The sporozoites of four Haemoproteus lineages were detected in Culicoides segnis, C. festivipennis, and Culicoides kibunensis biting midges. PCR-based screening showed that the females of seven Culicoides species were naturally infected with Haemoproteus parasites. The DNA of the parasite of owls, Haemoproteus syrnii (hSTAL2), was detected for the first time in Culicoides punctatus. The highest abundance of collected Culicoides females was in June, but the highest prevalence of Haemoproteus parasites in biting midges was in July. The abundance of Culicoides was higher in the woodland compared with reeds during the season. The acquired findings indicate the varied abundance and diversity of biting midges throughout the season and across distinct habitats. This variability could potentially impact the transmission of Haemoproteus parasites among birds with diverse breeding site ecologies. These outcomes hold the potential to enhance our understanding of the epizootiology of Haemoproteus infections within temperate climatic zones.

The article focuses on the rapid spread of bluetongue disease in northwestern Europe despite the introduction of new vaccines. Topics include the disease's resurgence in the Netherlands and its spread to other European countries, the limited effectiveness of the new vaccines, and the significant economic and agricultural impacts of the outbreak.

The biting midge Culicoides circumscriptus Kieffer, 1918 is a European widespread vector of avian malaria throughout the continent and is a possible vector of Akabane virus and Bluetongue virus. This species populates a wide range of environments in contrasting ecological settings often exposed to strong seasonal fluctuations. The main goals of this study were to investigate C. circumscriptus phenotypic variation at three departments in France (Corsica Island, Moselle and Var) and to determine if its phenotypes vary with the environment. Culicoides circumscriptus wing phenotypes were analyzed using a geometric morphometric approach based on anatomical landmarks and outlines of the wing. Dendogram trees based on landmarks and the outlines-2 set (cell m4) showed similar topologies and separated populations of C. circumscriptus. In contrast, another set of outlines-1 (covering the r-m cross vein, M, radiale and arculus) presented a different hierarchical clustering tree. The phenotypic variation observed in C. circumscriptus indicated that these populations are exposed to environmental and ecological pressures. Our results suggest the presence of phenotypic plasticity in this species.

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

A winter population of around 4000–5000 wild Eurasian tundra reindeer (Rangifer t. tarandus) in the eastern part of Iceland represents descendants from 35 semi-domesticated reindeer imported to Iceland from Finnmark county, Norway, in 1787. While previous studies have indicated that they host fewer parasite species as compared to reindeer in Fennoscandia, little information exists on their exposure to reindeer viral pathogens. The aim of this study was to investigate blood from hunted reindeer for antibodies against alphaherpesvirus and gammaherpesviruses (malignant catarrhal fever viruses, MCFV), pestivirus, bluetongue virus, and Schmallenberg virus, and to investigate nasal and oral mucosal membrane swab samples for the presence of parapoxvirus-specific DNA. Blood samples collected during the hunting seasons in 2017 (n = 40), 2018 (n = 103), and 2019 (n = 138) were tested for viral antibodies using enzyme-linked immunosorbent assays (ELISA). Screening for parapoxvirus DNA was conducted on swab samples from 181 reindeer by polymerase chain reaction (PCR), targeting the B2L and GIF genes. Antibodies against pestivirus were detected in two animals from 2017, and antibodies against MCFV were detected in two reindeer from 2018. No antibodies were detected against the other viruses tested. Parapoxvirus-specific DNA was detected in nasal swab samples from two animals sampled in 2019. This study suggests that the investigated viral infections are either not present or present at a low prevalence only, probably not representing a major health threat to this reindeer population. The lack of exposure to alphaherpesvirus, an enzootic pathogen in most investigated Rangifer populations, was unexpected. [ABSTRACT FROM AUTHOR]

Bluetongue (BT), once considered a disease of sheep confined to the southern African region, has spread all over the world. BT is a viral disease caused by the bluetongue virus (BTV). BT is regarded as an economically important disease in ruminants of compulsory notification to OIE. BTV is transmitted by the bite of Culicoides species. Research over the years has led to a better understanding of the disease, the nature of the virus life cycle between ruminants and Culicoides species, and its distribution in different geographical regions. Advances have also been made in understanding the molecular structure and function of the virus, the biology of the Culicoides species, its ability to transmit the disease, and the persistence of the virus inside the Culicoides and the mammalian hosts. Global climate change has enabled the colonization of new habitats and the spread of the virus into additional species of the Culicoides vector. This review highlights some of the current findings on the status of BT in the world based on the latest research on disease aspects, virus-host-vector interactions, and the different diagnostic approaches and control strategies available for BTV. [ABSTRACT FROM AUTHOR]

Vector-borne diseases constitute a major global public health threat. The most significant arthropod disease vectors are predominantly comprised of members of the insect order Diptera (true flies), which have long been the focus of research into host–pathogen dynamics. Recent studies have revealed the underappreciated diversity and function of dipteran-associated gut microbial communities, with important implications for dipteran physiology, ecology, and pathogen transmission. However, the effective parameterization of these aspects into epidemiological models will require a comprehensive study of microbe-dipteran interactions across vectors and related species. Here, we synthesize recent research into microbial communities associated with major families of dipteran vectors and highlight the importance of development and expansion of experimentally tractable models across Diptera towards understanding the functional roles of the gut microbiota in modulating disease transmission. We then posit why further study of these and other dipteran insects is not only essential to a comprehensive understanding of how to integrate vector-microbiota interactions into existing epidemiological frameworks, but our understanding of the ecology and evolution of animal-microbe symbiosis more broadly. [ABSTRACT FROM AUTHOR]

Culcioides biting midges (Diptera: Ceratopogonidae) are vectors of various veterinary pathogens. Suction light traps are one of the most widely used tools for vector surveillance. The present aim was to compare the efficiency for the collection of Culicoides species between newly developed 3D-printed ultraviolet (Mahidol University (MU) UV LED) and green light-emitting diode (Mahidol University (MU) Green LED) traps baited with CO2 and UV LED Center for Disease Control (CDC) light trap (BioQuip 2770) baited with CO2. The experiment consisted of two replicates of a 3 × 3 Latin square design in each three sampling locations (Location 1, 2, 3 and 4, 5, 6), for 12 nights between 26th July and 7th August 2020 in Thailand. Results showed that efficiency of the MU UV LED light trap was equivalent to that of the BioQuip 2770 trap for the collection of Culicoides. Meanwhile, the efficiency of the MU Green LED light trap was lower than that of both UV LED light traps. In the analysis of Culicoides species composition and sex–age grading, a similar pattern was observed among three light traps except for Culicoides actoni Smith. The newly developed 3D-printed UV LED light trap demonstrated the following advantages over the commercial light trap: cost saving to obtain multiple units, ease of customization and standardization, and increased availability by end-users. Although further assessments in different environmental conditions are needed, this 3D-printed light trap design could minimize the constrains in vector surveillance programs worldwide. [ABSTRACT FROM AUTHOR]

Background: Molecular analysis of blood meals is increasingly used to identify the hosts of biting insects such as midges and mosquitoes. Successful host identification depends on the availability of sufficient host DNA template for PCR amplification, making it important to understand how amplification success changes under different storage conditions and with different durations of blood meal digestion within the insect gut before being placed into the storage medium. Method: We characterised and compared the digestion profile of two species of Culicoides over a 96-h period using a novel set of general vertebrate primers targeting the 16S rRNA gene. A set number of individuals from each species were killed over 13 time points post-blood feeding and preserved in 95% ethanol. Samples were stored either at ambient room temperature or in a − 20 °C freezer to examine the effect of storage condition on the PCR amplification success of host DNA. Results: We found that amplification success across the 96-h sampling period post-feeding was reduced from 96 to 6% and 96% to 14% for Culicoides nubeculosus and Culicoides sonorensis, respectively. We found no effect of storage condition on PCR amplification success, and storage in 95% ethanol was sufficient to maintain high rates of amplifiable host DNA for at least 9 months, even at room temperature. Conclusions: These findings highlight the limited time frame during which an individual may contain amplifiable host DNA and demonstrate the importance of timely sample capture and processing post-blood feeding. Moreover, storage in 95% ethanol alone is sufficient to limit host DNA degradation. These results are relevant to the design of studies investigating the biting behaviour and disease transmission potential of Culicoides and other biting Diptera. [ABSTRACT FROM AUTHOR]

Determination of host preferences of blood-feeding arthropods is an important criterion for how ectoparasite-host interactions, host selection, and feeding behaviour affects pathogen transmission. This study aimed to examine the host's blood from engorged Culicoides species collected from cattle farms in Turkey to shed light on their host preferences. For this purpose, we investigated the blood of 5 different hosts (cattle, sheep, dog, horse, and human) in 7 Culicoides species/species complexes by multiplex-PCR analysis, considering the domestic animals on the farms. Engorged Culicoides specimens were collected from places in Turkey with different geographical and ecological characteristics. A total of 1225 female Culicoides belonging to 7 different species/species complexes namely C. obsoletus complex (C. obsoletus and C. scoticus) (n:450), C. schultzei complex (n:234), C. imicola (n:208), C. punctatus (n:162), C. newsteadi (n:144), C. lupicaris (n:24), and C. pulicaris (n:3) were analysed for host blood identification. Abdomens of the engorged midges were separated from their body and pooled according to the date of collection and species. A total of 69 pools consisting of 1-28 specimens were analysed by multiplex-PCR and only cattle blood was detected in all pools. This study presents the first data on the identification of host preference of some Culicoides species in Turkey. [ABSTRACT FROM AUTHOR]

Epizootic haemorrhagic disease (EHD) is a Culicoides-borne viral disease caused by the epizootic haemorrhagic disease virus (EHDV) associated with clinical manifestations in domestic and wild ruminants, primarily white-tailed deer (Odocoileus virginianus) and cattle (Bos taurus). In late September 2021, EHDV was reported in cattle farms in central/western Tunisia. It rapidly spread throughout the country with more than 200 confirmed outbreaks. We applied a combination of classical and molecular techniques to characterize the causative virus as a member of the serotype EHDV-8. This is the first evidence of EHDV- 8 circulation since 1982 when the prototype EHDV-8 strain was isolated in Australia. This work highlights the urgent need for vaccines for a range of EHDV serotypes. [ABSTRACT FROM AUTHOR]

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

Vector-borne disease prevalence is increasing at a time when surveillance capacity in the United States is decreasing. One way to address this surveillance deficiency is to utilize established infrastructure, such as zoological parks, to investigate animal disease outbreaks and improve our epidemiological understanding of vector-borne pathogens. During fall 2020, an outbreak of epizootic hemorrhagic disease (EHD) at the Minnesota Zoo resulted in morbidity and seroconversion of several collection animals. In response to this outbreak, insect surveillance was conducted, and the collected insects were tested for the presence of epizootic hemorrhagic disease virus (EHDV) by RT-qPCR to better understand the local transmitting vector populations responsible for the outbreak. Six pools of Culicoides biting midges were positive for EHDV, including three pools of Culicoides sonorensis, two pools of Culicoides variipennis, and a pool of degraded C. variipennis complex midges. All three endemic serotypes of EHDV (1, 2, and 6) were detected in both animals and midge pools from the premises. Despite this outbreak, no EHDV cases had been reported in wild animals near the zoo. This highlights the importance and utility of using animal holding facilities, such as zoos, as sentinels to better understand the spatio-temporal dynamics of pathogen transmission. [ABSTRACT FROM AUTHOR]

Bluetongue (BT) is an acute, viral, vector-borne, non-contagious disease of ruminants caused by Bluetonguevirus (BTV). Twenty-six different serotypes have been identified, and the ability of each strain to cause disease varies widely. BTV can infect a wide range of domestic and wild ruminants - sheep, goats, cattle, buffalo, deer, most species of African antelope and camels. For the study period (2011-2021), outbreaks were declared in 21 member states in Europe with predominant serotypes 1, 4 and 8. Based on data from Animal Disease Information System (ADIS) with the present paper we analyzed the distribution of the BTV serotypes within the affected countries and found that particular serotypes tend to spread to particular countries. The development of regulatory measures for prevention, surveillance and eradication of BT is discussed based on the European legislation. [ABSTRACT FROM AUTHOR]

BACKGROUND: Sudden outbreaks of the orange wheat blossom midge, Sitodiplosis mosellana (Géhin) cause huge wheat yield losses. Use of sex pheromones is more efficient than laborious egg counting to monitor these hidden‐concealed insects. Quick synthesis of the sex pheromones is therefore required to meet the sudden outbreak needs. RESULTS: A synthetic approach of stereospecific and racemic S. mosellana pheromones was presented. This method afforded the stereospecific and racemic S. mosellana pheromones in three steps and high enantioselectivity (> 98% ee for (2S,7S)‐2,7‐nonanediyl dibutyrate) in less than 1 day with 74% and 73% overall yields, respectively, whereas most conventional methods require longer synthesis time with less than 40% yield. The synthesis routes could quickly and economically afford the pheromones, starting from synthon (S)‐but‐3‐yn‐2‐ol (1a) or but‐3‐yn‐2‐ol (1b), through the same three‐step processes of coupling, reduction, and esterification. The Y‐tube olfactometer results showed significant attractiveness of the synthetic stereospecific and racemic sex pheromones to S. mosellana males relative to the blank control (P < 0.001). Field trials also demonstrated significant attractiveness of the synthetic stereospecific and racemic sex pheromones relative to the blank control (P < 0.001). CONCLUSION: This modular approach is conducive to the deployment of field traps and timely responses to S. mosellana outbreaks and can be a time‐saving and cost‐effective tool to manage S. mosellana. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Two new Brazilian species of Monohelea Kieffer are described and illustrated based on male specimens, Monohelea capixaba sp. nov. from Espírito Santo and Monohelea coimbrai sp. nov. from Rio de Janeiro. New records for M. aguirrei Tavares & Souza, M. archibaldoi Tavares & Souza and M. maculipennis (Coquillet) are given based on specimens from Espírito Santo (all three species) and Amapá (M. maculipennis only). All specimens are deposited in the Ceratopogonidae Collection of Fundação Oswaldo Cruz, Brazil. [ABSTRACT FROM AUTHOR]

The female of the biting midge Forcipomyia paludis is a dipteran ectoparasite of West Palaearctic damselflies and dragonflies, sucking haemolymph mainly from wing veins of their hosts. This tiny midge remains firmly attached to the wings even during fast flight and aerial fight maneuvers as shown in the present paper by field studies of the large dragonfly, Cordulegaster boltonii. Since individuals of F. paludis firmly attach themselves to the challenging wing surface of their host and can successfully withstand drag and vibrations during flight, we assume that this midge species has specific microstructural adaptations on its legs for attaching to the wing surface. In our morphological study, we used scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), to study the structure of F. paludis tarsi, as well as the micro morphology of the wing surfaces of their host. Additionally, for the first time, we were able to show attachment devices of the midges dried out in contact with the host's surface. The spatulae of the plantar setae and especially the empodial setae, are capable of replicating nanoscale wax crystals of the super hydrophobic wing coverage of the dragonfly wing membrane, in order to increase an effective contact area and therefore adhesion. This ability requires extremely soft materials of the spatula, which seems to be rather unique even in comparison to the leg attachment devices of other dipterans and other insect taxa in general. [ABSTRACT FROM AUTHOR]

The new species Atrichopogon (Atrichopogon) tolfensisn. sp. from Tolfa Mountains (Italy, Latium, and Rome) is described and illustrated. Both sexes of the species were collected using cantharidin-baited traps from June 2020 to January 2021. The new species is an atypical member of the genus Atrichopogon. [ABSTRACT FROM AUTHOR]

Culicoides biting midges (Diptera: Ceratopogonidae) are biting nuisances and arbovirus vectors of both public health and veterinary significance in Trinidad. We compared sampling methods to define the behaviour and bionomics of adult Culicoides populations at a commercial dairy goat farm. Three static trap designs were compared: (a) Centre for Disease Control (CDC) downdraft UV trap; (b) CDC trap with an incandescent bulb and (c) CDC trap with semiochemical lure consisting of R‐(−)‐1‐octen‐3‐ol and CO2 (no bulb). Sweep netting was used to define diel periodicity. A total of 30,701 biting midges were collected using static traps, dominated by female Culicoides furens (>70% of trap collections across all three designs). There was no significant difference in the Margalef's index between the three traps; however, trap designs A and C collected a significantly greater number of individuals than trap B, and trap C gained highest species richness. The greatest species richness and abundance of Culicoides collected by sweep net was observed between 6:00 and 6:15 pm and notable differences in the crepuscular activity pattern of several species were identified. Comparative data on Culicoides species richness, abundance, sex and reproductive status is discussed and can be used to improve surveillance strategies, research designs and risk management. [ABSTRACT FROM AUTHOR]

Knowledge gaps exist on the feeding pattern and host range of bluetongue virus vectors, Culicoides species, associated with livestock in India. Adult midges were trapped with ultraviolet light traps at 13 household farms adjacent to human biotope. Host DNA was isolated from individual females (n = 101; blood engorged‐82, gravid‐4 and parous‐15) and subjected to PCR amplification targeting CytB and 16S rRNA gene fragments followed by sequencing of amplified DNA samples. However, DNA sequences from only 71 individuals (70.3%) comprising of 10 Culicoides species were obtained. Blood meal analysis revealed at least 10 species that fed on five mammalian hosts including humans, but surprisingly none tested positive for birds. Results revealed that Culicoides innoxius tested positive for four not previously recognized species indicating a potential role as a vector species. Likewise, Culicoides shortti and Culicoides hegneri preferred goat and cattle respectively as hosts, whereas Culicoides palpifer preferred cattle along with buffalo as hosts, which is being reported for the first time. This is the first document on DNA‐based blood meal identification and feeding preference of Culicoides midges associated with livestock in India. [ABSTRACT FROM AUTHOR]

The diversity and abundance of Diptera at the Agri-biodiversity Park, Professor Jayashankar Telangana State Agricultural University, Hyderabad was studied from September 2019 to January 2020. Five collection methods (pitfall trap, yellow pan trap, manual collection, light trap and yellow sticky trap) were used and 6739 individuals under 27 families were collected. The results revealed that the most abundant family was Dolichopodidae (10.76%) followed by Sarcophagidae (10.19%) and Ceratopogonidae (8.92%). Yellow pan trap collected maximum number of individuals (2789), and revealed occurrence of maximum number of families (18). Shannon-Weiner diversity Index, Margalef’s species richness index and Pielou’s evenness index of the study area were 2.953, 2.949 and 0.896, respectively, indicating rich diversity of Diptera. [ABSTRACT FROM AUTHOR]

Abstract Background The Middle East Respiratory Syndrome coronavirus (MERS‐CoV) is still listed on the WHO Research and Development Blueprint of emerging pathogens. Dromedary camels remain the only known animal reservoir of the virus. The animal‐to‐animal as well as the animal‐to‐human transmission in the MERS‐CoV cycles were reported. However, many aspects of these transmission chains are not well studied. One of these directions is the potential roles of various species of arthropods in the transmission of the virus. Objectives The main goal of the current work was to study the roles of several species of arthropods in the transmission of MERS‐CoV. Methodology To achieve this goal, we identified some MERS‐CoV naturally infected dromedary camel populations. We conducted a longitudinal study among these animals for more than 2 months. This was done by repeated testing of nasal swabs biweekly from some selected animals in this population for the presence of MERS‐CoV‐RNAs by real‐time PCR. During the duration of this study, we collected several species of arthropods (Culicoides, Stomoxys, Musca domestica and some Culex species) that shared the habitat and were circulating in this farm during this longitudinal study. Results Our results showing, despite the detection of the viral RNAs in some animals throughout this study, none of the examined species of arthropods tested positive for the viral RNA. Conclusions These results are suggesting that at least the tested species of arthropods may not play roles in the transmission of MERS‐CoV. However, more large‐scale studies are required to explore any potential roles of arthropods in the transmission cycle of MERS‐CoV.

Abstract Background Bluetongue (BT), an infectious, non-contagious arthropod borne viral disease of domestic and wild ruminants is caused by bluetongue virus (BTV), a type species of Orbivirus genus and Reoviridae family. Albeit bluetongue in India is endemic to southern and western parts, eastern and north-eastern parts of the country didn’t experience any outbreak or reported active disease. Main body of abstract International, national and regional disease statuses including some un-published data were narrated to give a clear picture. Until 1940s, BT was mainly restricted to African countries, however started spearheading to Asian and European countries subsequently. Extensive epidemiological surveys were conducted in central parts of America and Caribbean countries during late 80’s. Massive ingression of BT was observed in northern Europe 2006 onward. In India, although first reported in 1964, BT became endemic in southern states during 1980s and became alarming in late 1990s. A thorough sero-epidemiological study was conducted in different agro-climatic zones of eastern and north-eastern states of India during 2001 to 2017. Sero-epidemiological information of different states had been tabulated. This was the first comprehensive document of BT sero-epidemiology of eastern and north-eastern parts of India. Although BT was not reported officially, sero-epidemiology showed BTV was in circulation in animals and vectors of eastern and north-eastern states of India. Published as also un-published data clearly showed animals and vectors harbor BTV in this part of the country. Short conclusion All the epidemiological components of BT are present in eastern and north-eastern parts of India that warrants presence of the disease in these parts also. Hence, further surveillance and preparedness are recommended to avoid future BT outbreak, if any.

Bluetongue virus (BT) is a vector-borne virus that causes a disease, called bluetongue, which results in significant economic loss and morbidity in sheep, cattle, goats and wild ungulates across all continents of the world except Antarctica. Despite the geographical breadth of its impact, most BT epidemiological models are informed by parameters derived from the 2006–2009 BTV-8 European outbreak. The aim of this study was to develop a highly adaptable model for BT which could be used elsewhere in the world, as well as to identify the parameters which most influence outbreak dynamics, so that policy makers can be properly informed with the most current information to aid in disease planning. To provide a framework for future outbreak modelling and an updated parameterisation that reflects natural variation in infections, a newly developed and parameterised two-host, two-vector species ordinary differential equation model was formulated and analysed. The model was designed to be adaptable to be implemented in any region of the world and able to model both epidemic and endemic scenarios. It was parameterised using a systematic literature review of host-to-vector and vector-to-host transmission rates, host latent periods, host infectious periods, and vaccine protection factors. The model was demonstrated using the updated parameters, with South Africa as a setting based on the Western Cape's known cattle and sheep populations, local environmental parameters, and Culicoides spp. presence data. The sensitivity analysis identified that the duration of the infectious period for sheep and cows had the greatest impact on the outbreak length and number of animals infected at the peak of the outbreak. Transmission rates from cows and sheep to C. imicola midges greatly influenced the day on which the peak of the outbreak occurred, along with the duration of incubation period, and infectious period for cows. Finally, the protection factor of the vaccine had the greatest influence on the total number of animals infected. This knowledge could aid in the development of control measures. Due to gradual climate and anthropological change resulting in alterations in vector habitat suitability, BT outbreaks are likely to continue to increase in range and frequency. Therefore, this research provides an updated BT modelling framework for future outbreaks around the world to explore transmission, outbreak dynamics and control measures. [ABSTRACT FROM AUTHOR]

Mansonella perstans infections are widespread in Sub-Saharan Africa and Central and South America and thus can be considered as the most prevalent parasite of man in tropical Africa. In contrast to the high prevalence, knowledge about the biology of this filarial nematode is restricted and no effective treatment regimens of this ivermectin-resistant parasite is lacking. An obstacle for the research is that M. perstans resides in body cavities and thus have been only rarely recovered during surgery or autopsy. Therefore, alternative methods like in vitro culture systems need to be implemented to decipher the nature of mansonellosis and effective drugs. Previously, we have established a monkey kidney epithelial cell-based in vitro culture for the maintenance of M. perstans infective larvae (L3) up to 77 days. However, no alternative for this culture system have been postulated to allow longer survival rates and development of adult worms in vitro. Thus, we aim to establish an alternative in vitro culture system for M. perstans L3. M. perstans L3 were isolated from engorged and laboratory reared Culicoides midges. The larvae were then cultured in Dulbecco's Modified Eagle Medium supplemented with either 10% foetal bovine serum (FBS), 10% newborn calf serum (NCS) or 1% bovine serum albumin (BSA) together with human colon carcinoma cells (HCT-8) as feeder cells. Survival and growth were recorded. We obtained that the 10% NCS culture condition was superior allowing long-term maintenance of M. perstans L3 for up to 100 days and boosted growth of the parasites for up to 5-folds compared to the initial size at culture inception. Although no moulting of the L3 into L4 or adult worms could be overserved, the human colon carcinoma cell-based in vitro culture provides an alternative platform to analyse M. perstans biology and screen for novel drugs against M. perstans. [Display omitted] • Mansonella perstans is considered as the most prevalent parasite of man in tropical Africa. • In vitro culture systems for long-term maintenance of M. perstans are mandatory. • Human colon carcinoma cell-based culture allows parasite survival. • Addition of 10% newborn calf serum allow parasite survival and growth for up to 100 days. [ABSTRACT FROM AUTHOR]

• The emergence of EHDV-8 in Europe deserves quick countermeasures to mitigate its impact on livestock industry. • An inactivated vaccine against EHDV-8 was developed and tested in cattle. • The vaccine was safe and induced a robust neutralizing immune response. • Vaccinated calves were fully protected from clinical EHD and the development of viraemia following challenge. Epizootic haemorrhagic disease (EHD), caused by the EHD virus (EHDV), is a vector-borne viral disease transmitted through Culicoides biting midges. EHDV comprises seven serotypes (1, 2, and 4–8), with EHDV-8 having recently emerged and spread in Europe over the last two years. Such event has raised concerns about the significant threat posed by EHDV-8 to livestock industry. In this study, an inactivated vaccine against EHDV-8 (vEHDV8-IZSAM) was developed. Safety and efficacy of the vaccine were evaluated in calves through clinical, serological, and virological monitoring following experimental challenge. The vaccine was proven safe, with only transient fever and localized reactions observed in a few animals, consistent with adjuvanted vaccine side effects. vEHDV8-IZSAM elicited a robust humoral response, as evidenced by the presence of neutralizing antibodies. After challenge with a virulent isolate, viraemia and clinical signs were evidenced in control animals but in none of the vaccinated animals. This study highlights the potential of vEHDV8-IZSAM as a safe and highly effective vaccine against EHDV-8 in cattle. It offers protection from clinical disease and effectively prevents viraemia. With the recent spread of EHDV-8 in European livestock, the use of an inactivated vaccine could be key in protecting animals from clinical disease and thus to mitigate the economic impact of the disease. Further investigations are warranted to assess the duration of the induced immunity and the applicability of this vaccine in real-world settings. Accordingly, joint efforts between public veterinary institutions and pharmaceutical companies are recommended to scale up vaccine production. [ABSTRACT FROM AUTHOR]

Bluetongue disease is an infectious disease transmitted by Culicoides as vectors, mainly infecting ruminants. Because ruminants play an important role in animal husbandry in China, the outbreak of bluetongue disease can cause serious economic losses. Maxent model was applied to predict the distribution of bluetongue in China based on the data derived from domestic and foreign academic literature databases including CNKI, Wanfang Database, PubMed, Web of Science and Google Scholar. The results showed that annual mean temperature (BIO1), precipitation in driest month (BIO14), sheep density (SD) and altitude (Elev) were the relevant variables of bioclimatic suitable zones for bluetongue disease. Precipitation in wettest month (BIO13), BIO1, BIO14, Elev were the main variables affecting the habitat of the bluetongue vector Culicoides. The most suitable climate for bluetongue infection occurs in southern China, central China and parts of Xinjiang. The suitable living areas of Culicoides are mainly located in southern, central and eastern China, and the overlap of the two suitable areas is high. The study suggested that southern, central, and eastern China are high-risk areas for bluetongue due to the significant overlap of suitable habitats for both the disease and its vector. Implementing effective surveillance and targeted control strategies in these regions is crucial for mitigating the impact of bluetongue disease. [ABSTRACT FROM AUTHOR]

Biting midges of the genus Culicoides occur almost globally and can regionally and seasonally reach high abundances. Most species are hematophagous, feeding on all groups of vertebrates, including humans. In addition to being nuisance pests, they are able to transmit disease agents, with some viruses causing high morbidity and/or mortality in ruminants, horses and humans. Despite their impact on animal husbandry, public health and tourism, knowledge on the biology and ecology of culicoid biting midges and their interactions with ingested pathogens or symbiotic microorganisms is limited. Research is challenging due to unknown larval habitats, the insects’ tiny size, the inability to establish and breed most species in the laboratory and the laborious maintenance of colonies of the few species that can be reared in the laboratory. Consequently, the natural transmission of pathogens has experimentally been demonstrated for few species while, for others, only indirect evidence of vector potential exists. Most experimental data are available for Culicoides sonorensis and C. nubeculosus, the only species kept in western-world insectaries. This contribution gives an overview on important biting midge vectors, transmitted viruses, culicoid-borne viral diseases and their epidemiologies and summarizes the little knowledge on interactions between biting midges, their microflora and culicoid-borne arboviruses.

Viruses that are transmitted by arthropods, or arboviruses, have evolved to successfully navigate both the invertebrate and vertebrate hosts, including their immune systems. Biting midges transmit several arboviruses including vesicular stomatitis virus (VSV). To study the interaction between VSV and midges, we characterized the transcriptomic responses of VSV-infected and mock-infected Culicoides sonorensis cells at 1, 8, 24, and 96 h post inoculation (HPI). The transcriptomic response of VSV-infected cells at 1 HPI was significant, but by 8 HPI there were no detectable differences between the transcriptome profiles of VSV-infected and mock-infected cells. Several genes involved in immunity were upregulated (ATG2B and TRAF4) or downregulated (SMAD6 and TOLL7) in VSV-treated cells at 1 HPI. These results indicate that VSV infection in midge cells produces an early immune response that quickly wanes, giving insight into in vivo C. sonorensis VSV tolerance that may underlie their permissiveness as vectors for this virus.

Bluetongue virus (BTV) is a segmented, double-stranded RNA virus transmitted by Culicoides biting midges. Infection of domestic and wild ruminants with BTV can result in a devastating disease and significant economic losses. As a virus with a segmented genome, reassortment among the BTV serotypes that have co-infected a host may increase genetic diversity, which can alter BTV transmission dynamics and generate epizootic events. The objective of this study was to determine the extent of dissemination and characterize the tropism of BTV serotypes 10 and 17 in co-infected Culicoides sonorensis. Midges were exposed to both BTV serotypes via blood meal and processed for histologic slides 10 days after infection. An in situ hybridization approach was employed using the RNAscope platform to detect the nucleic acid segment 2 of both serotypes. Observations of the mosaic patterns in which serotypes did not often overlap suggest that co-infection at the cellular level may not be abundant with these two serotypes in C. sonorensis. This could be a consequence of superinfection exclusion. Understanding BTV co-infection and its biological consequences will add an important dimension to the modeling of viral evolution and emergence.

ABSTRACT Mosquitoes and biting Culicoides species are arbovirus vectors. Effective virome profile surveillance is essential for the prevention and control of insect-borne diseases. From June to September 2021, we collected eight species of female mosquito and Culicoides on Zhoushan Island, China, and used meta-viromic sequencing to analyze their virome compositions and characteristics. The classified virus reads were distributed in 191 genera in 66 families. The virus sequences in mosquitoes with the largest proportions were Iflaviridae (30.03%), Phasmaviridae (23.09%), Xinmoviridae (21.82%), Flaviviridae (13.44%), and Rhabdoviridae (8.40%). Single-strand RNA+ viruses formed the largest proportions of viruses in all samples. Blood meals indicated that blood-sucking mosquito hosts were mainly chicken, duck, pig, and human, broadly consistent with the habitats where the mosquitoes were collected. Novel viruses of the Orthobunyavirus, Narnavirus, and Iflavirus genera were found in Culicoides by de-novo assembly. The viruses with vertebrate hosts carried by mosquitoes and Culicoides also varied widely. The analysis of unclassified viruses and deep-learning analysis of the “dark matter” in the meta-viromic sequencing data revealed the presence of a large number of unknown viruses. IMPORTANCE The monitoring of the viromes of mosquitoes and Culicoides, widely distributed arbovirus transmission vectors, is crucial to evaluate the risk of infectious disease transmission. In this study, the compositions of the viromes of mosquitoes and Culicoides on Zhoushan Island varied widely and were related mainly to the host species, with different host species having different core viromes. and many unknown sequences in the Culicoides viromes remain to be annotated, suggesting the presence of a large number of unknown viruses.

Tibet orbivirus (TIBOV), a new candidate of Orbivirus genus, was initially isolated from mosquitoes in Tibet in 2009 and subsequently from both Culicoides and mosquitoes in several provinces of China and Japan. Little is known about the origin, genetic diversity, dissemination and pathogenicity of TIBOV, although its potential threat to animal health has been acknowledged. In this study, two viruses, V290/YNSZ and V298/YNJH, were isolated from the Culicoides and sentinel cattle in Yunnan Province. Their genome sequences, cell tropism in mammalian and insect cell lines along with pathogenicity in suckling mice were determined. Genome phylogenetic analyses confirmed their classification as TIBOV species; however, OC1 proteins of the V290/YNSZ and V298/YNJH shared maximum sequence identities of 31.5% and 33.9% with other recognized TIBOV serotypes (TIBOV‐1 to TIBOV‐4) and formed two monophyletic branches in phylogenetic tree, indicating they represented two novel TIBOV serotypes which were tentatively designated as TIBOV‐5 and TIBOV‐6. The viruses replicated robustly in BHK, Vero and C6/36 cells and triggered overt clinical symptoms in suckling mice after intracerebral inoculation, causing mortality of 100% and 25%. Cross‐sectional epidemiology analysis revealed silent circulation of TIBOV in Yunnan Province with overall prevalence of 16.4% (18/110) in cattle, 10.8% (13/120) in goats and 5.5% (6/110) in swine. The prevalence patterns of four investigated TIBOV serotypes (TIBOV‐1, ‐2, ‐5 and 6) differed from each one another, with their positive rates ranging from 8.2% (9/110) for TIBOV‐2 in cattle to 0.9% (1/110) for TIBOV‐1 and TIBOV‐5 in cattle and swine. Our findings provided new insights for diversity, pathogenicity and epidemiology of TIBOV and formed a basis for future studies addressing the geographical distribution and the zoonotic potential of TIBOV. [ABSTRACT FROM AUTHOR]

The territory of the Tyumen region is located in the West Siberian Lowland and is characterized by conditions favorable for the development of blood-sucking diptera insects. The fauna of the blood-sucking diptera insects of the Tyumen region is represented by 109 species and 4 subspecies: horseflies-35 species and 4 subspecies, mosquitoes-41 species, blackflies-16 species, biting midges-17 species. In the subzone of the southern taiga, Hybomitra ciureai, H. bimaculata, H. lundbecki, H. muehlfeldi, Haematopota pluvialis, Aedes cantans, A. punctor, Simulium maculatum, Culicoides punctatus dominate; in the subzone of aspenbirch forests-Tabanus bromius, Hybomitra ciureai, H. bimaculata, H. lundbecki, Atylotus fulvus, A. rusticus, Haematopota subcylindrica, H. pluvialis, Aedes cinereus, A. punctor, A. cantans, Simulium pusillum, Culicoides punctatus; in the forest-steppe zone-Tabanus bovinus, T. bromius, Hybomitra ciureai, H. bimaculata, H. lundbecki, Haematopota pluvialis, H. subcylindrica, Coquillettidia richiardii, Aedes caspius, A. albescens, A. cantans, Simulium maculatum, Culicoides punctatus. The total summer period of insects of this group ranges from 140 days in the subzone of the southern taiga to 170 days in the forest-steppe zone. Depending on the meteorological conditions of the spring-summer season, the start and end dates of summer may shift by 1-2 decades. The activity of horseflies and blackflies is observed during the daytime, and mosquitoes and biting midges are twilight insects and attack mainly in the morning and evening hours. The main abiotic factors determining the nature of the diurnal rhythm of the vital activity of diptera insects in the conditions of the Tyumen region are air temperature and illumination. [ABSTRACT FROM AUTHOR]

The rapid and disorderly urbanization in the Amazon has resulted in the insertion of forest fragments into cities, causing the circulation of arboviruses, which can involve hematophagous arthropods and free-ranging birds in the transmission cycles in urban environments. This study aimed to evaluate the circulation of arboviruses in free-ranging birds and hematophagous arthropods captured in an Environmental Protection Area in the Belem metropolitan area, Brazil. Birds were captured using mist nets, and hematophagous arthropods were collected using a human protected attraction technique and light traps. The birds' sera were subjected to a hemagglutination inhibition test to detect antibodies against 29 arbovirus antigens. Arthropod macerates were inoculated into C6/36 and VERO cell cultures to attempt viral isolation and were tested using indirect immunofluorescence, subsequent genetic sequencing and submitted for phylogenetic analysis. Four bird sera were positive for arbovirus, and one batch of Psorophora ferox was positive for Flavivirus on viral isolation and indirect immunofluorescence. In addition, the Ilheus virus was detected in the sequencing and phylogenetic analysis. The presence of antibodies in sera from free-ranging birds and the isolation of Ilheus virus in Psorophora ferox indicate the circulation of arboviruses in forest remnants in the urban center of Belem. [ABSTRACT FROM AUTHOR]