Insights from common buzzard broods on the interaction between Leucocytozoon infection, watercourse habitats and simuliid blackfly vectors.

Blood parasites of the genus Leucocytozoon commonly infect many bird species worldwide and are particularly prevalent in birds of prey. As a vector-borne parasitic disease, the infection occurrence overlaps with that of the dominant vectors: blackflies (Diptera, Simuliidae). These blood-sucking insects are dependent on habitats with flowing freshwaters for the development of their larval stages. We investigated the correlation between the proximity to flowing waters and Leucocytozoon infection probability in common buzzard (Buteo buteo) broods, as well as the occurrence of adult blackflies directly at the nests. In addition, we investigated the survival of captured simuliids in relation to host infection intensity. In total in 2019, we examined 112 different nests, including 297 common buzzard nestlings, with a Leucocytozoon prevalence of 56.6% among the nestlings and of 80.3% at brood level. We found no significant association of Leucocytozoon infection probability with nestling age, the distance to the nearest stream and the sum of the length of streams within a radius of 200 and 1000 m around each nest. The number of blackflies caught around the nest showed a tentative correlation with the probability of Leucocyozoon infection of the nestlings. Among the subsample of 218 blackfly individuals that survived day one after capture, survival averaged 6.2 days. Our results suggest that Leucocytozoon transmission is complex and requires consideration of many factors, related to habitat and vector prevalence, especially given their temporal variation.

Morphological description and genetic analysis of a new black fly species (Diptera: Simuliidae) in the subgenus Asiosimulium from central Thailand.

BACKGROUND: Black flies are among the most medically and veterinary important insects, as adult females of certain species are the sole vector of Onchocerca volvulus. Here, a new black fly species belonging to the subgenus Asiosimulium Takaoka & Choochote, 2005, is described and formally named as Simulium (Asiosimulium) kittipati sp. nov. METHODS: Pupae and larvae of black flies were collected from available substrates in the stream from central Thailand. Pupae were individually separated in plastic tubes and maintained until adult flies emerged. The emerged adult flies associated with their pupal exuviae and cocoon as well as mature larvae preserved in 85% ethanol were used to describe the new species based on an integrated approach of morphological examination and molecular analysis of the COI gene. RESULTS: The new species is characterized in the female by the medium-long sensory vesicle with a medium-sized opening apically, scutum with three faint longitudinal vittae, and the ellipsoidal spermatheca; in the male by the number of upper-eye (large) facets in 20 vertical columns and 21 horizontal rows, hind basitarsus slender, nearly parallel-sided, and median sclerite much wider and upturned apically; in the pupa by the head and thoracic integument densely covered with tiny tubercles, and the pupal gill of arborescent type with 28-30 filaments; and in the larva by the postgenal cleft deep, nearly reaching the posterior margin of the hypostoma, and dark pigmented sheath of the subesophageal ganglion. The DNA barcode successfully differentiated the new species from its congeners with an interspecific genetic divergence of 1.74-18.72%, confirming the morphological identification that the species is a new member of the subgenus Asiosimulium. Phylogenetic analyses also indicated that the new species is genetically closely related to Simulium phurueaense Tangkawanit, Wongpakam & Pramual, 2018, further supporting its morphological classification. CONCLUSIONS: This is the ninth species assigned to the subgenus Asiosimulium within the genus Simulium Latreille, 1802. Taxonomic notes and identification keys are given to distinguish this new species from the eight known species members in its same subgenus. Additionally, a distribution map of all species members in this subgenus occurring in Thailand and other countries is provided.

Ratio changes in blood-feeding and voltinism in black flies (Diptera: Simuliidae) over a latitudinal gradient in North America north of Mexico.

We explored how the ratio of blood-feeding behaviour-nonfeeding, bird feeding (ornithophily) and mammal feeding (mammalophily)-and voltinism (univoltine and multivoltine) vary over a latitudinal gradient from Alaska to Florida. These two fixed species traits were divided into five mutually exclusive combinations of trait states-nonfeeding/univoltine, ornithophilic/univoltine, mammalophilic/univoltine, ornithophilic/multivoltine and mammalophilic/multivoltine-within each of three datasets (north, east and west). We found a significant association between location (north, east and west) and trait state, which was driven by the large percentage of nonfeeding females in the north. When this trait state was removed, no significant differences were found for the remaining trait states and locations. Although the distribution of trait states did not differ between east and west datasets, the distribution with relation to 1° changes in latitude within each of these datasets showed distinct differences. In the east, both ornithophilic/univoltine and mammalophilic/univoltine species significantly increased with latitude, in proportion to the total species present. In contrast, the proportion of mammalophilic/multivoltine species decreased as latitude increased. Ornithophilic/multivoltine species in the east and the trait states in the west did not show any significant relationship to latitude.

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.

The possible role of haematophagous flies in the incidence of bovine teat papillomatosis.

The relationship between the incidence of bovine teat papillomatosis and the activity of haematophagous flies was investigated in Japan. A total of 15,737 flies consisting of 33 species were collected by dry ice-baited mosquito net (DMN) trap and a sweep net from udders of cattle. Simulium aokii (Takahasi) of Simuliidae (black flies) was the predominant species, followed by S. tobetsuense Ono and S. iwatense (Shiraki). Simulium aokii had the highest peak in October, followed by September. Numbers of blood spots from the bites per teat in nulliparous cattle were significantly correlated with numbers of S. aokii collected by DMN trap. Numbers of teats with warts and spots of blood from the bites per teat were significantly more abundant in anterior teats than posterior teats. The average incidence of teat papillomatosis in nulliparous cattle was significantly higher than that in parous cattle, and the highest incidence by month was in May, followed by April. Although bovine papillomavirus (BPV) DNA was not detected in flies examined, the presence of black flies and blood spots from their bites were associated with subsequent high incidence of growing warts. In particular, it would pay to give attention to species such as S. aokii that severely attack udders in the present locality. Further investigations for the detection of BPV DNA from flies parasitizing on teats are needed.

Community composition of black flies during and after the 2020 vesicular stomatitis virus outbreak in Southern New Mexico, USA.

BACKGROUND: Vesicular stomatitis virus (VSV), a vector-borne pathogen of livestock, emerges periodically in the western US. In New Mexico (NM), US, most cases occur close to the Rio Grande River, implicating black flies (Simulium spp.) as a possible vector. In 2020, VS cases were reported in NM from April to May, although total black fly abundance remained high until September. We investigated the hypothesis that transience of local VSV transmission results from transient abundance of key, competent black fly species. Additionally, we investigated whether irrigation canals in southern NM support a different community of black flies than the main river. Lastly, to gain insight into the source of local black flies, in 2023 we collected black fly larvae prior to the release of water into the Rio Grande River channel. METHODS: We randomly sub-sampled adult black flies collected along the Rio Grande during and after the 2020 VSV outbreak. We also collected black fly adults along the river in 2021 and 2022 and at southern NM farms and irrigation canals in 2022. Black fly larvae were collected from dams in the area in 2023. All collections were counted, and individual specimens were subjected to molecular barcoding for species identification. RESULTS: DNA barcoding of adult black flies detected four species in 2020: Simulium meridionale (N = 158), S. mediovittatum (N = 83), S. robynae (N = 26) and S. griseum/notatum (N = 1). Simulium robynae was only detected during the VSV outbreak period, S. meridionale showed higher relative abundance, but lower absolute abundance, during the outbreak than post-outbreak period, and S. mediovittatum was rare during the outbreak period but predominated later in the summer. In 2022, relative abundance of black fly species did not differ significantly between the Rio Grande sites and farm and irrigation canals. Intriguingly, 63 larval black flies comprised 56% Simulium vittatum, 43% S. argus and 1% S. encisoi species that were either extremely rare or not detected in previous adult collections. CONCLUSIONS: Our results suggest that S. robynae and S. meridionale could be shaping patterns of VSV transmission in southern NM. Thus, field studies of the source of these species as well as vector competence studies are warranted.

Ecologically and medically important black flies of the genus Simulium: Identification of biogeographical groups according to similar larval niches.

The black fly genus Simulium includes medically and ecologically important species, characterized by a wide variation of ecological niches largely determining their distributional patterns. In a rapidly changing environment, species-specific niche characteristics determine whether a species benefits or not. With aquatic egg, larval and pupal stages followed by a terrestrial adult phase, their spatial arrangements depend upon the interplay of aquatic conditions and climatic-landscape parameters in the terrestrial realm. The aim of this study was to enhance the understanding of the distributional patterns among Simulium species and their ecological drivers. In an ecological niche modelling approach, we focused on 12 common black fly species with different ecological requirements. Our modelling was based on available distribution data along with five stream variables describing the climatic, land-cover, and topographic conditions of river catchments. The modelled freshwater habitat suitability was spatially interpolated to derive an estimate of the adult black flies' probability of occurrence. Based on similarities in the spatial patterns of modelled habitat suitability we were able to identify three biogeographical groups, which allows us to confirm old assessments with current occurrence data: (A) montane species, (B) broad range species and (C) lowland species. The five veterinary and human medical relevant species Simulium equinum, S. erythrocephalum, S. lineatum, S. ornatum and S. reptans are mainly classified in the lowland species group. In the course of climatic changes, it is expected that biocoenosis will slightly shift towards upstream regions, so that the lowland group will presumably emerge as the winner. This is mainly explained by wider ecological niches, including a higher temperature tolerance and tolerance to various pollutants. In conclusion, these findings have significant implications for human and animal health. As exposure to relevant Simulium species increases, it becomes imperative to remain vigilant, particularly in investigating the potential transmission of pathogens.

First record of the mermithid nematode worm Isomermis lairdi parasitizing black flies in Spain.

Mermithid nematodes are considered a promising biological control agent to reduce the population density of different blood-feeding vectors, i.e. black flies (Diptera: Simuliidae), which are important pests of medical and veterinary interest worldwide. Immature larvae of black flies were collected in a rill from La Rioja (Northern Spain) in the summer of 2016. Isomermis lairdi Mondet, Poinar & Bernadou, 1977 (Nematoda: Mermithidae) was found parasitizing eleven specimens of Simulium cryophilum s.l. (Rubtsov, 1959) (prevalence of 52%), which represent the first record of this nematode for Spain and the second for Europe. The confirmation of the nematode and the black fly species was carried out by both morphological and molecular approaches using the 18S ribosomal RNA and the cytochrome c oxidase subunit I genes. Phylogenetic analyses indicated that the collected specimens were Isomermis lairdi (99.4-99.9% identity with homologues from Africa) with a sequence divergence of 0.2%. The role of Isomermis lairdi as an alternative tool in the biological control of black flies in Spain should be further explored.

Occurrence and Characterization of Small Microplastics (<100 µm), Additives, and Plasticizers in Larvae of Simuliidae

This study is the first to investigate the ingestion of microplastics (MPs), plasticizers, additives, and particles of micro-litter < 100 µm by larvae of Simuliidae (Diptera) in rivers. Blackflies belong to a small cosmopolitan insect family whose larvae are present alongside river courses, often with a torrential regime, up to their mouths. Specimens of two species of blackfly larvae, Simulium equinum and Simulium ornatum, were collected in two rivers in Central Italy, the Mignone and the Treja. Small microplastics (SMPs, <100 µm), plasticizers, additives, and other micro-litter components, e.g., natural and non-plastic synthetic fibers (APFs) ingested by blackfly larvae were, for the first time, quantified and concurrently identified via MicroFTIR. The pretreatment allowed for simultaneous extraction of the ingested SMPs and APFs. Strong acids or strong oxidizing reagents and the application of temperatures well above the glass transition temperature of polyamide 6 and 6.6 (55−60 °C) were not employed to avoid further denaturation/degradation of polymers and underestimating the quantification. Reagent and procedural blanks did not show any SMPs or APFs. The method's yield was >90%. Differences in the abundances of the SMPs and APFs ingested by the two species under exam were statistically significant. Additives and plasticizers can be specific to a particular polymer; thus, these compounds can be proxies for the presence of plastic polymers in the environment.

Survey and Phylogenetic Analysis of Leucocytozoon (Apicomplexa: Haemosporida) Parasites in Mississippi Black Flies (Diptera: Simuliidae).

For several years, there have been continuous reports of black flies in Mississippi with evidence of transmission of Leucocytozoon spp. This study was conducted to determine the presence and diversity of Leucocytozoon spp. in black flies captured throughout Mississippi. Thirty-three collections, consisting of 346 specimens, were made during the 2-yr period (2015-2016) at 10 locations around the state. In addition to these systematic biweekly collections, 46 specimens were taken in 10 collections at four additional sites from 2009 to 2014, as well as 388 specimens taken in 14 collections during a severe black fly outbreak during March and April of 2018. Out of 186 pooled samples, 21 samples were positive for haemosporidian DNA. Eighteen of those samples were identified as Leucocytozoon spp. A phylogenetic tree was constructed using the novel sequences along with existing sequences from the MalAvi database. Results showed several of the sequences constructed in this study had high divergence from the existing sequences from the database.

Environmental drivers of breeding sites in blackfly species of medical and veterinary importance in eastern Spain.

Geographical distribution and abundance of the pupae of six blackfly species of medical and veterinary concern were studied in eastern Spain according to three different sets of explanatory variables including in-stream variables, both (i) abiotic (i.e., physicochemical) and (ii) biotic (i.e., richness and abundance of either taxonomically or ecologically close related taxa), as well as (iii) meteorological and landscape variables. The results showed specific habitat requirements for pupation in Simulium (Boophthora) erythrocephalum (De Geer, 1776) and Simulium (Wilhelmia) equinum (Linnaeus, 1758), two of the six species studied regarding elevation and temperature. While the rest of the species showed a certain degree of ecological overlap, co-occurrence was in general low, which suggested that antagonistic biotic factors may be important in structuring blackfly assemblages. In effect, biotic predictors explained a high proportion (50%-70%) of the variability in the abundance of the pupae of the most generalist blackfly species, although further studies are needed to disentangle the sign of interspecific interactions. At the landscape level, S. (W.) equinum and S. (W.) pseudequinum Séguy, 1921 breeding habitats were associated with the presence of pig farms, and S. (Simulium) reptans (Linnaeus, 1758) and S. (B.) erythrocephalum with the presence of cattle.

Descriptions of two new species of the Simulium (Simulium) striatum species-group (Diptera: Simuliidae) from Taiwan and Japan, and a revised description of S. (S.) quinquestriatum (Shiraki).

Species in the Simulium (Simulium) striatum species-group (Diptera: Simuliidae) in Taiwan and Japan are morphologically reevaluated. Simulium (S.) yanpingense sp. nov. is described as the third member in this species-group from Taiwan and a revised description of S. (S.) quinquestriatum (Shiraki) is provided. The species previously regarded as S. (S.) quinquestriatum in Japan is described as a new species, S. (S.) tanegashimaense. Simulium (S.) yanpingense sp. nov. is similar to S. (S.) pingtungense Huang & Takaoka from Taiwan, in many characteristics including the haired basal portion of the female radius, dark leg color, and male scutum with brassy hairs, but it is distinguished from the latter species by the number of male upper-eye (large) facets, which are in 17 (rarely 18) vertical columns and 17 or 18 horizontal rows in this new species but in 19 vertical columns and 20 horizontal rows in S. (S.) pingtungense. Simulium (S.) tanegashimaense sp. nov. is similar to S. (S.) quinquestriatum originally described from Taiwan by having the male scutum with yellow fine hairs, but is distinguished from the latter species by the number of male upper-eye (large) facets, which are in 17 (rarely 16) vertical columns and 17 horizontal rows in this new species but in 19 or 20 vertical columns and 19 or 20 horizontal rows in S. (S.) quinquestriatum. The morphological differences between these two new species and their related species from India, Malaysia, Thailand and Vietnam are noted.

Surveillance along the Rio Grande during the 2020 Vesicular Stomatitis Outbreak Reveals Spatio-Temporal Dynamics of and Viral RNA Detection in Black Flies.

Vesicular stomatitis virus (VSV) emerges periodically from its focus of endemic transmission in southern Mexico to cause epizootics in livestock in the US. The ecology of VSV involves a diverse, but largely undefined, repertoire of potential reservoir hosts and invertebrate vectors. As part of a larger program to decipher VSV transmission, we conducted a study of the spatiotemporal dynamics of Simulium black flies, a known vector of VSV, along the Rio Grande in southern New Mexico, USA from March to December 2020. Serendipitously, the index case of VSV-Indiana (VSIV) in the USA in 2020 occurred at a central point of our study. Black flies appeared soon after the release of the Rio Grande's water from an upstream dam in March 2020. Two-month and one-year lagged precipitation, maximum temperature, and vegetation greenness, measured as Normalized Difference Vegetation Index (NDVI), were associated with increased black fly abundance. We detected VSIV RNA in 11 pools comprising five black fly species using rRT-PCR; five pools yielded a VSIV sequence. To our knowledge, this is the first detection of VSV in the western US from vectors that were not collected on premises with infected domestic animals.

Morphological and genetic revision of cytoform 'L' of the Simulium (Simulium) tani complex from Taiwan, and cytoform 'D' of the S. (S.) suzukii complex (Diptera: Simuliidae) from the Nansei Islands, Japan.

The species status of two cytoforms of two species complexes in the Simulium (Simulium) tuberosum species-group, which are almost indistinguishable in the pupal stage, was morphologically and genetically evaluated. Cytoform 'L' of the S. (S.) tani Takaoka & Davies complex, previously recognized as S. (S.) suzukii Rubtsov in Taiwan, is described as a new species, S. (S.) jianshiense. It is morphologically distinguishable as adults from two members of the S. (S.) suzukii complex: cytoform 'C' from Hokkaido, selected to represent the type of S. (S.) suzukii sensu stricto, and cytoform 'D' from Okinawa Island and, based on our chromosomal analysis, also from Amami Island. This new species is genetically separated from both cytoforms of the S. (S.) suzukii complex with a genetic distance of 5.31-6.67%. Cytoform 'D' is distinguished from cytoform 'C' by the color of the male forecoxa and relative length of the female sensory vesicle to the third maxillary palpomere. Accordingly, the name S. (S.) ryukyuense Ogata, once regarded as a synonym of S. (S.) suzukii, is revalidated and applied to cytoform 'D'. The genetic distance between S. (S.) ryukyuense and S. (S.) suzukii sensu stricto is 1.24-1.60%.

DNA barcoding of tropical black flies (Diptera: Simuliidae) in Thailand: One decade of progress.

Black flies (Diptera: Simuliidae) are important blood sucking insects because they are the vectors of disease agents transmitted to human and other animals. Rapid and correct species identifications are necessary for all aspects of the study of black flies. DNA barcodes based on mitochondrial cytochrome c oxidase I (COI) have been effectively used for the determination of black fly species. However, the success of this method requires a large and reliable COI sequence library. In this study, 171 DNA barcoding sequences from 17 black fly species were added to NCBI GenBank database, six of these species were reported for the first time. Efficacy of DNA barcodes for species identification was examined using 1,286 sequences representing 89 nominal species of black flies in Thailand. A considerable level of success (90%) was achieved but efficiency of COI sequences for species identification was very low in the following species-groups; Simulium asakoae, S. feuerborni, S. multistriatum and S. striatum. Incomplete lineage sorting or inadequate variation of this genetic marker for differentiation of recently diverged species are the more likely explanations, and thus, more variable genetic markers are needed. Other reasons for unsuccessful DNA barcoding are imperfect taxonomy and the misidentification of sources of reference sequences. Because many new black fly species in Thailand were described recently, a reassessment of the COI sequences reported previously is necessary.

Community Structure, Biodiversity and Spatiotemporal Distribution of the Black Flies (Diptera: Simuliidae) Using Malaise Traps on the Highest Mountain in Thailand.

Black flies form a group of small blood-sucking insects of medical and veterinary importance. This study aimed to investigate the community structure, biodiversity and spatial and temporal distribution of adult black flies in tropical rain forests, by using malaise traps in Doi Inthanon National Park, northern Thailand. Malaise traps were placed along six elevational gradients (400 m to 2500 m, above sea level) at Doi Inthanon National Park, Chiang Mai province, from December 2013 to November 2014. A total of 9406 adult female black flies belonging to five subgenera-Daviesellum (2%), Gomphostilbia (23%), Montisimulium (11%), Nevermannia (16%) and Simulium (48%)-were collected. Among 44 taxa found, S. tenebrosum complex had the highest relative abundance (11.1%), followed by the S. asakoae species-group (9.6%), the S. striatum species-group (7.7%), S. inthanonense (6.6%), S. doipuiense complex (6.4%), S. chomthongense complex (5.3%), S. chumpornense (5.1%) and S. nigrogilvum (4.1%). Two human-biting species-S. nigrogilvum and species in the S. asakoae species-group-were found in all of the collection sites with 100% species occurrence. Species richness was highest at mid elevation (1400 m), which is represented by 19 black fly species. The peak and lowest seasonal abundance was observed in the rainy and hot season, respectively. Seasonal species richness was highest in the cold season, except for that from elevation sites at 700 m, 1700 m and 2500 m. This study revealed that the malaise trap is effective in providing important data for further monitoring of the effects of environmental changes and conservation planning on the biodiversity of black flies in Doi Inthanon National Park.

Atypical behavior of a black fly species connects cavity-nesting birds with generalist blood parasites in an arid area of Spain.

BACKGROUND: The feeding behavior of bloodsucking insects determines the transmission, distribution, host spectrum and evolution of blood parasites in the wild. Conventional wisdom suggests that some vector groups (e.g. black flies, family Simuliidae) are consistently exophagous daytime biters. We aimed to understand more about the exceptions to this pattern by combining targeted trapping and molecular identification of parasites in vectors. METHODS: In this study, we collected black flies in nest boxes used by European rollers Coracias garrulus in southeastern Spain. We molecularly analyzed 434 individual insects, identifying the black fly species caught in the nest boxes, their potential vertebrate blood meals, and the haemosporidian parasite lineages that they carried. RESULTS: Only one black fly species, Simulium rubzovianum, appeared to enter the nest boxes of rollers. Among the trapped specimens, 15% contained vertebrate DNA, which always belonged to rollers, even though only half of those specimens were visibly engorged. Furthermore, 15% of all black flies contained Leucocytozoon lineages, indicating previous feeding on avian hosts but probably not on infected adult rollers. The known vertebrate hosts of the recorded Leucocytozoon lineages suggested that large and/or abundant birds are their hosts. Particularly represented were cavity-nesting species breeding in the vicinity, such as pigeons, corvids and owls. Open-nesting species such as thrushes and birds of prey were also represented. CONCLUSIONS: Our data strongly suggest that S. rubzovianum bites uninfected roller nestlings and infected individuals of other species, potentially incubating adults, inside nest boxes and natural cavities. This simuliid does not appear to have a strong preference for specific host clades. Contrary to the general pattern for the group, and possibly enhanced by the harsh environmental conditions in the study area, this black fly appeared to intensively use and may even have a preference for confined spaces such as cavities for feeding and resting. Preferences of vectors for atypical microhabitat niches where hosts are less mobile may enable social and within-family transmission and parasite speciation in the long term. At the same time, a lack of host preference in concentrated multispecies communities can lead to host switches. Both processes may be underappreciated driving forces in the evolution of avian blood parasites.

Environmental and Spatial Predictors of the Distribution Patterns of the Host-Seeking Black Fly, Simulium jenningsi (Diptera: Simuliidae).

Black flies are nuisance pests as adults, yet they are best managed in the larval stage in flowing waters. As a result, more effort is put into understanding the distribution of the immature life stages than the blood-seeking females that form nuisance. The seemingly localized nature of Simulium jenningsi Malloch (Diptera: Simuliidae) pest problems in western Maryland offered a study system to investigate the spatial and environmental correlates to their severity. Collections of adult black flies were taken at 260 sites within a 2,000 km2 region centered on Washington County, Maryland, during June, July, and August of 2 yr. Average S. jenningsi counts were greater in the June of both years compared to July and August. Although S. jenningsi was found at the majority of sampling sites, higher fly counts were significantly clustered in the southern portion of the county where the majority of resident complaints originated. A generalized linear mixed-model (GLMM) approach was used to determine the correlates to S. jenningsi abundance. The highest performing model showed a negative relationship of S. jenningsi counts with the amount of surrounding impervious surface, distance to the riffles along the confluence of the Shenandoah and Potomac Rivers, distance to the closest body of flowing water, and light intensity, as well as a positive relationship with elevation and air temperature. The results suggest S. jenningsi females are not readily found in urban environments in this study region, and the most relevant monitoring locations for S. jenningsi may be outside of human population centers.

Simulium (Gomphostilbia) aziruni: First record of a black fly (Diptera: Simuliidae) attracted to a human in Malaysia.

Most female black flies in the genus Simulium are blood-sucking flies and they can cause various parasitic diseases in human and animal. A total of 94 species of black flies have been reported in Malaysia, however, their biting behavior and role as vector of infectious agents remain understudied. To fill in this knowledge gap, we attempted to survey adult black flies from field populations in Peninsular Malaysia. In a survey carried out in 2017 at Tasik Kenyir, Terengganu, three females were caught while attracted and landed on human skin. Further morphological and molecular analyses showed that the specimens were identical to Simulium (Gomphostilbia) aziruni Takaoka, Hashim & Chen of the Simulium gombakense species-group. This is the first report on a black fly species attracted to human in Malaysia which serves as a steppingstone towards in-depth studies for black flies in this region.

Skin Lesions and Systemic Reactions in Humans Infested by Blackflies (Diptera: Simullidae) in Recreational Areas in Southeastern Poland.

Due to their mass occurrence in some environments and high aggressiveness, blackflies (Simulium spp.) represent the most bothersome arthropods attacking humans. In this study, we describe the medical effects of blackfly infestations in humans in southeastern Poland. Local and systemic reactions to blackfly bites were monitored in 418 patients (61.24% of females and 38.76% of males) of medical centers. Only skin lesions at the site of the bites were found in 88.52% of the patients, whereas accompanying systemic reactions were diagnosed in 11.48%. The most common signs observed in the area of the bites were pruritus (94.74%), burning (55.02%), edema (44.02%), and erythema (40.91%). The skin lesions, which were most often grouped small papules and papular and purpuric lesions with a varied range, typically persisted for several days, or for several weeks in some patients. Statistical analyses confirmed that the persistence of the skin lesions did not depend on the sex of the patients and the number of blackfly infestations. The systemic reactions to the components of the blackfly saliva were manifested by headache, increased body temperature, arthralgia, lymphadenopathy, and menstrual disorders in the females. The patients were most often attacked simultaneously by many blackflies on exposed parts of the body, mainly the upper limb, lower limb, head, and neck areas. The investigations indicate that blackflies are an important etiological factor of dermatitis and other symptoms in humans; hence, they should be considered in clinical diagnosis.