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Simulium (Simulium) phraense sp. nov. (Diptera: Simuliidae) is described from females, males, pupae, and larvae from Thailand. This new species is placed in the Simulium striatum species group and is most similar to Simulium (Simulium) nakhonenseTakaoka & Suzuki (Diptera: Simuliidae) from Thailand among species of the same species group but is barely distinguished from the latter species by lacking annular ridges on the surface of the pupal gill filaments.The fast-evolving nuclear big zinc finger (BZF) gene has successfully differentiated this new species from its allies, S. (S.) nakhonense and Simulium (Simulium) chiangmaiense Takaoka & Suzuki (Diptera: Simuliidae) of the S. striatum species group.The BZF gene sequences show that this new species is more closely related to S. (S.) nakhonense than to S. (S.) chiangmaiense, further supporting its morphological classification.

Background Fast and reliable species identification of black flies is essential for research proposes and effective vector control. Besides traditional identification based on morphology, which is usually supplemented with molecular methods, geometric morphometrics (GM) has emerged as a promising tool for identification. Despite its potential, no specific GM techniques have been established for the identification of black fly species. Methods Adult female black flies collected using human bait, as well as those reared from pupae, were used in this study. Here, landmark-based GM analysis of wings was assessed for the first time to identify human-biting black fly species in Thailand, comparing this approach with the standard morphological identification method and DNA barcoding based on the mitochondrial cytochrome c oxidase subunit I ( COI ) gene. To explore genetic relationships between species, maximum likelihood (ML) and neighbor-joining (NJ) phylogenetic trees were built. Additionally, three different methods of species delimitation, i.e., assemble species by automatic partitioning (ASAP), generalized mixed yule coalescent (GMYC), and single Poisson tree processes (PTP), were utilized to identify the morphologically defined species. The effectiveness of a COI barcode in identifying black fly species was further examined through the best match (BM) and best close match (BCM) methods. Results Seven black fly species, namely Simulium tenebrosum Takaoka, Srisuka & Saeung, 2018 (complex), S. doipuiense Takaoka & Choochote, 2005 (complex), S. nigrogilvum Summers, 1911, S. nodosum Puri, 1933, S. asakoae Takaoka & Davies, 1995, S. chamlongi Takaoka & Suzuki, 1984, and S. umphangense Takaoka, Srisuka & Saeung, 2017 were morphologically identified. Compared with the standard method, the GM analysis based on wing shape showed high success in separating species, achieving an overall accuracy rate of 88.54%. On the other hand, DNA barcoding surpassed wing GM for species identification with a correct identification rate of 98.57%. Species delimitation analyses confirmed the validity of most nominal species, with an exception for S. tenebrosum complex and S. doipuiense complex, being delimited as a single species. Moreover, the analyses unveiled hidden diversity within S. asakoae , indicating the possible existence of up to four putative species. Conclusions This study highlights the potential of wing GM as a promising and reliable complementary tool for species identification of human-biting black flies in Thailand. Graphical Abstract

In Africa, livestock production currently accounts for about 30% of the gross value of agricultural production. However, production is struggling to keep up with the demands of expanding human populations, the rise in urbanization and the associated shifts in diet habits. High costs of feed prevent the livestock sector from thriving and to meet the rising demand. Insects have been identified as potential alternatives to the conventionally used protein sources in livestock feed due to their rich nutrients content and the fact that they can be reared on organic side streams. Substrates derived from organic by-products are suitable for industrial large-scale production of insect meal. Thus, a holistic comparison of the nutritive value of Black Soldier Fly larvae (BSFL) reared on three different organic substrates, i.e. chicken manure (CM), brewers’ spent grain (SG) and kitchen waste (KW), was conducted. BSFL samples reared on every substrate were collected for chemical analysis after the feeding process. Five-hundred (500) neonatal BSFL were placed in 23 × 15 cm metallic trays on the respective substrates for a period of 3–4 weeks at 28 ± 2 °C and 65 ± 5% relative humidity. The larvae were harvested when the prepupal stage was reached using a 5 mm mesh size sieve. A sample of 200 grams prepupae was taken from each replicate and pooled for every substrate and then frozen at −20 °C for chemical analysis. Samples of BSFL and substrates were analyzed for dry matter (DM), crude protein (CP), ether extracts (EE), ash, acid detergent fibre (ADF), neutral detergent fibre (NDF), amino acids (AA), fatty acids (FA), vitamins, flavonoids, minerals and aflatoxins. The data were then subjected to analysis of variance (ANOVA) using general linear model procedure. BSFL differed in terms of nutrient composition depending on the organic substrates they were reared on. CP, EE, minerals, amino acids, ADF and NDF but not vitamins were affected by the different rearing substrates. BSFL fed on different substrates exhibited different accumulation patterns of minerals, with CM resulting in the largest turnover of minerals. Low concentrations of heavy metals (cadmium and lead) were detected in the BSFL, but no traces of aflatoxins were found. In conclusion, it is possible to take advantage of the readily available organic waste streams in Kenya to produce nutrient-rich BSFL-derived feed.

Efforts to recycle organic wastes using black soldier fly (BSF) Hermetia illucens into high-nutrient biomass that constitutes a sustainable fat (biodiesel) and high-quality protein ingredient in animal feeds have recently gained momentum worldwide. However, there is little information on the most suitable rearing conditions for growth, development and survivorship of these flies, which is a prerequisite for mass production technologies. We evaluated the physiological requirements for growth and reproduction of H. illucens on two diets [spent grains supplemented with brewers' yeast (D1) and un-supplemented (D2)]. Development rates at nine constant temperatures (10-42°C) were fitted to temperature-dependent linear and non-linear day-degree models. Thereafter, life history table parameters were determined within a range of favourable temperatures. The thermal maximum (TM) estimates for larval, pre-pupal and pupal development using non-linear model ranged between 37.2 ± 0.3 and 44.0 ± 2.3°C. The non-linear and linear day-degree model estimations of lower developmental temperature threshold for larvae were 11.7 ± 0.9 and 12.3 ± 1.4°C for D1, and 10.4 ± 1.7 and 11.7 ± 3.0°C for D2, respectively. The estimated thermal constant of immature life stages development of BSF was higher for the larval stage (250±25 DD for D1 and 333±51 for D2) than the other stages evaluated. Final larval wet weight was higher on D1 compared to D2. The population growth rate was most favourable at 30-degree celsius (°C) with higher intrinsic rate of natural increase (rm = 0.127 for D1 and 0.122 for D2) and shorter doubling time (5.5 days for D1 and 5.7 days for D2) compared to the other temperatures. These results are valuable for the optimization of commercial mass rearing procedures of BSF under various environmental conditions and prediction of population dynamics patterns using computer simulation models.

The widespread nominal black fly Simulium (Simulium) rufibasis Brunetti was reexamined morphologically, chromosomally, and molecularly to determine the status of populations in Japan and Korea with respect to S. rufibasis from the type locality in India and to all other known species in the S. (S.) tuberosum species-group. Morphological comparisons established that the species previously known as S. rufibasis in Japan and Korea is distinct from all other species. Consequently, it was described and illustrated as a new species, Simulium (S.) yamatoense. Simulium yokotense Shiraki, formerly a synonym of S. rufibasis, was morphologically reevaluated and considered a species unplaced to species-group in the subgenus Simulium. Chromosomal analyses of S. yamatoense sp. nov. demonstrated that it is unique among all cytologically known species of the S. tuberosum group and is the sister species of the Taiwanese species tentatively known as S. (S.) arisanum Shiraki. Populations of S. yamatoense sp. nov. included two cytoforms, based on the sex chromosomes. Cytoform A, including topotypical representatives, was found in Kyushu, Japan, whereas cytoform B was found in Korea and Honshu, Japan. Molecular analysis based on the COI mitochondrial gene generally corroborated morphological and chromosomal data that S. yamatoense sp. nov. is a distinct species and, like the chromosomal data, indicate that it is most closely related to S. arisanum, with interspecific genetic distance of 2.92–4.63%.

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. Graphical abstract

Background The onchocerciasis focus surrounding the lower Mbam and Sanaga rivers, where Onchocerca volvulus is transmitted by Simulium damnosum s.l. (Diptera: Simuliidae), was historically the largest in the southern regions of Cameroon. Annual community-directed treatment with ivermectin (CDTI) has been taking place since 2000, but recent studies have shown that new infections are occurring in children. We aimed to investigate blackfly biting and O. volvulus transmission rates along the lower Mbam river 16 years after the formal onset of annual CDTI. Methods Black flies were collected for three consecutive days each month between July 2016 and June 2017 at two riverside villages and two inland sites situated 4.9 km and 7.9 km from the riverside. Specimens collected at each site were dissected on one of the three collection days each month to estimate parity rates and O. volvulus infection rates, while the remaining samples were preserved for pool screening. Results In total, 93,573 S. damnosum s.l. black flies were recorded biting humans and 9281 were dissected. Annual biting rates of up to 606,370 were estimated at the riverside, decreasing to 20,540 at 7.9 km, while, based on dissections, annual transmission potentials of up to 4488 were estimated at the riverside, decreasing to 102 and 0 at 4.9 km and 7.9 km, respectively. However, pool screening showed evidence of infection in black flies at the furthest distance from the river. Results of both methods demonstrated the percentage of infective flies to be relatively low (0.10–0.36%), but above the WHO threshold for interruption of transmission. In addition, a small number of larvae collected during the dry season revealed the presence of Simulium squamosum E. This is the first time S. squamosum E has been found east of Lake Volta in Ghana, but our material was chromosomally distinctive, and we call it S. squamosum E2. Conclusions Relatively low O. volvulus infection rates appear to be offset by extremely high densities of biting black flies which are sustaining transmission along the banks of the lower Mbam river. Graphical Abstract

The species richness of black flies in the Simulium (Trichodagmia) orbitale (Diptera: Simuliidae) species group is greatest in southern Brazil, where 9 of 19 species are found. A new species in the S. orbitale group was collected during a survey of black flies in Minas Gerais state, Brazil, in the Espinhaço Mountains, an area rich in endemic taxa. The new species is morphologically similar to Simulium guianense Wise (Diptera: Simuliidae), the main vector of the causal agent of onchocerciasis in Brazil and Venezuela. It can be identified in the male by the unpatterned scutum and slightly concave ventral plate with a prominent median projection, in the female by the bilobate anal lobe, in the pupa by the 12 slender gill filaments with nonsclerotized apices, branching in a rake-like pattern, and in the larva by the elongated abdominal segments V–VIII and ventrolaterally lobulate segment IX. The polytene chromosomes have one unique fixed inversion in the IS arm, relative to all other Brazilian members of the group, confirming the morphological evidence of a distinct species.

The black fly Simulium (Trichodagmia) hirtipupa Lutz (Diptera: Simuliidae) is widely distributed in southern Brazil, with one report from Amapá state in the northern region of Brazilian Amazonia. Morphological comparison of northern and southern populations revealed differences in all life stages, corroborated by chromosomal and molecular analyses, and indicated that the population previously identified as S. hirtipupa from Amapá state represents an undescribed species. This new species is described based on all life stages above the egg, and its chromosomal and molecular divergence from S. hirtipupa is highlighted. Simulium criniferum n. sp. can be diagnosed by the deeply concave male ventral plate with a prominent median projection bearing a ventral keel; female anal lobe in lateral view with a broadly rounded, distal membranous area about as long as wide; pupa with a boot-shaped cocoon bearing a minutely bubbled surface, cephalic plate and thorax with abundant hair-like tubercles, and gill of 12 translucent filaments with darkly sclerotized, acuminate tips; and larva with the body cuticle bearing spiniform setae, abdomen truncated posteriorly, and gill histoblast in situ with the filament tips directed ventrally. Chromosomally, the new species has five unique fixed inversions and uniquely shares three additional fixed inversions with its nearest relative, S. hirtipupa. Partial COI sequences indicate a genetic distance of ∼9% between the new species and S. hirtipupa. Females of the new species are anthropophilic.

A new black fly species, Simulium haiduanenseTakaoka, Low & Huang (Diptera: Simuliidae), is described on the basis of females, males, pupae, and mature larvae from Taiwan. This new species is placed in the Simulium argentipes species-group of the subgenus Simulium (Diptera: Simuliidae) and is characterized by the yellowish female legs, ovipositor valves rounded apically and with its inner margin concave, claw with a small subbasal tooth, male style without a basal protuberance, pupal gill with eight filaments, corbicular cocoon, and larval abdomen lacking paired protuberances. It represents the first record of the S. argentipes species-group from Taiwan. Taxonomic notes are given to separate this new species from all eight species in the same species-group.The phylogenetic relationships of this new species with four related species are presented.