Dipteran Species Research Articles

Morphofunctional characterization of hemocytes in black soldier fly larvae.

In insects, the cell-mediated immune response involves an active role of hemocytes in phagocytosis, nodulation, and encapsulation. Although these processes have been well documented in multiple species belonging to different insect orders, information concerning the immune response, particularly the hemocyte types and their specific function in the black soldier fly Hermetia illucens, is still limited. This is a serious gap in knowledge given the high economic relevance of H.illucens larvae in waste management strategies and considering that the saprophagous feeding habits of this dipteran species have likely shaped its immune system to efficiently respond to infections. The present study represents the first detailed characterization of black soldier fly hemocytes and provides new insights into the cell-mediated immune response of this insect. In particular, in addition to prohemocytes, we identified five hemocyte types that mount the immune response in the larva, and analyzed their behavior, role, and morphofunctional changes in response to bacterial infection and injection of chromatographic beads. Our results demonstrate that the circulating phagocytes in black soldier fly larvae are plasmatocytes. These cells also take part in nodulation and encapsulation with granulocytes and lamellocyte-like cells, developing a starting core for nodule/capsule formation to remove/encapsulate large bacterial aggregates/pathogens from the hemolymph, respectively. These processes are supported by the release of melanin precursors from crystal cells and likely by mobilizing nutrient reserves in newly circulating adipohemocytes, which could thus trophically support other hemocytes during the immune response. Finally, the regulation of the cell-mediated immune response by eicosanoids was investigated.

A possible mechanism of Cry7Ab4 protein in delaying pupation of Plutella xylostella larvae.

Cry toxins produced by Bacillus thuringiensis (Bt) are well known for their insecticidal activities against Lepidopteran, Dipteran, and Coleopteran species. In our previous work, we showed that trypsin-digested full-length Cry7Ab4 protoxin did not have insecticidal activity against Plutella xylostella larvae but strongly inhibited their growth. In this paper, we expressed and purified recombinant active Cry7Ab4 toxic core from Escherichia coli for bioassay and identified its binding proteins. Interestingly, Cry7Ab4 toxic core exhibited activity to delay the pupation of P. xylostella larvae. Using protein pull-down assay, several proteins, including basic juvenile hormone-suppressible protein 1-like (BJSP-1), were identified from the midgut juice of P. xylostella larvae as putative Cry7Ab4-binding proteins. We showed that feeding P. xylostella larval Cry7Ab4 toxic core upregulated the level of BJSP-1 mRNA in the hemocytes and fat body and decreased the free juvenile hormone (JH) level in larvae. BJSP-1 interacted with Cry7Ab4 and bound to free JH in vitro. A possible mechanism of Cry7Ab4 in delaying the pupation of P. xylostella larvae was proposed.

Carrion flies (Insecta: Diptera) found on human cadavers in Chiba prefecture, Honshu, Japan, with the first record of Fannia prisca from a human corpse.

Forensic entomology (FE) involves the collection and analysis of necrophagous insects and arthropods for postmortem interval estimation based on their growth and succession. In Japan, research on FE is limited, and the predominant species of necrophagous insects and their distribution have not been clarified. The present study aimed to clarify the actual situation of insects, targeting Diptera collected from human cadavers, in Chiba Prefecture. We targeted the cases for which specimens could be collected from forensic autopsies conducted at the Legal Medicine Department of Chiba University Graduate School of Medicine or from human bodies handled by the Chiba Police in 2019. We specifically chose cases from which adult insects could be bred. Fifty cases were applicable, and 47 cases were successful in rearing adult flies. Sixteen species of Diptera were identified: six species of Calliphoridae, four species of Sarcophagidae, and a few species of Fanniidae, Muscidae, Phoridae, Piophilidae, and Stratiomyidae. The most frequently observed species were Lucilia sericata (20 cases) and Chrysomya pinguis (18 cases). For the first time, globally, we recorded the presence of Fannia prisca in a human cadaver. In conclusion, this is the first report to reveal the presence of carrion flies in human death cases in Chiba Prefecture, Japan. Research in FE is important in the context of the Japanese natural and social environment since this can aid forensic investigations conducted by authorities.

Managed pollination is a much better way of increasing productivity and essential oil content of dill seeds crop

Dill seeds (Anethumgraveolens L.) is the most valuable medicinal seed spice crop of Apiaceae. It bears small yellow flowers in the form of umbels. Being a cross-pollinated crop, floral visitors play vital role in pollination and seed sets. Hence, the present study was conducted at the ICAR-National Research Centre on Seed Spices, Ajmer (Rajasthan), India to discover the pollinator’s community, foraging behaviour and abundance of most frequent pollinators and different modes of pollination on seed yield and quality of this seed spice crop. The insect visitors community of dill seeds was composed of 28 insect species belonging to 14 families of 6 orders. Most of floral visitors started their foraging activity at 8.00 h, reached peak activity between 12.00 and 14.00 h and their activity ceased at 18.00 h. Apisflorea,A.dorsata,A.mellifera, solitary bee, Halictus sp. and two unidentified species of Hymenoptera; Episyrphusbalteatus (DeGeer), Episyrphus sp., Eristalis sp and two other Musca species of Diptera were identified as potential and regular floral visitors of dill seeds. The highest seed yield of 1505.63 kg/ha was recorded in the treated plots provided with only 10% jaggery solution and was at par with the open pollination. A lower seed yield of 1432.5 kg/ha was recorded in plots pollinated only with A.mellifera inside insect cages. Open pollination with 10% jaggery solution spray increased the seed yield of dill seed crop by 57%, one-thousand seed test weight by 96% and the essential oil content by 27% over control plots. These results show that managed pollination is a much better way to enhance yields and quality of dill seed crop than other treatments including only honeybee-based pollination.

Molecular Identification and Species Richness of Flies (Diptera) and Their Associated Bovidae Hosts at Cattle Farms in Selangor, Malaysia

Flies (Diptera) play a significant role in the ecosystem as pollinators and decomposers, and they are also important vermin and disease vectors. Studies on the dipteran species are still lacking in Malaysia; therefore, the dipteran species’ biology, morphology, distribution, and abundance are necessary. The objectives of this study were to identify dipteran species using a molecular approach, determine flies’ Bovidae hosts, and investigate the diversity of the fly’s species at three different cattle farms purposively selected in Selangor, Malaysia. The fly species were identified using cytochrome oxidase subunit I (COI) (Haematopota javana, Tabanus rubidus, Tabanus fontinalis, Iranihindia martellata, Musca domestica, and Chrysomya megacephala), while another six species only up to genus level ( Haematopota sp. 1, Musca sp. 1, Asilus sp., Metopia sp., Anasillomos sp., and Ommatius sp.). In addition, two species of hosts: Bos indicus and Bos taurus, were proven to be the associated host species for the dipteran species based on molecular data of cytochrome b (cytb). However, there were no significant differences between farms in species diversity and richness (F = 1.262, df = 2, p = 0.2459 >0.05). Interestingly, the two most dominant dipteran genera collected from the cattle farms were Musca and Chrysomya. At the same time, its abundance may have been influenced by the structure of the cattle cage flooring, which serves as a breeding site and food source. These findings contribute to fundamental epidemiological data in developing control strategies for dipteran species and are of great economic and health importance to livestock production in Malaysia.

Behaviour of two fly species reared for livestock feed: optimising production and insect welfare

The mass rearing of insects as animal feed is a new and rapidly growing component of circular agriculture, which offers the opportunity to develop it in such a way that it promotes insect health and welfare. Behaviour is an important indicator of animal performance and welfare. In this review, we synthesise the current behavioural knowledge on two saprophytic dipteran species that are increasingly being used as mini-livestock, the black soldier fly (Hermetia illucens) and the housefly (Musca domestica). We evaluate which behaviours need to be considered to optimise insect production and welfare under mass-rearing conditions. We distinguish between the different life stages (adults and larvae), and describe their feeding behaviour, social interactions (adult mating, larval aggregation), oviposition behaviour and possible cannibalism. For each species, we review what is known about these behaviours in natural environments, and how this is affected by abiotic factors or interactions with conspecifics and heterospecifics. We also address how the flies’ microbiome and pathogens can influence various aspects of behaviour. Notable differences in natural behaviours between the two species, such as their courtship and mating behaviour and the larval distribution within feed substrates are identified. These behavioural differences have important implications for how we should rear the two fly species in industrial settings, as a mismatch in mass-rearing conditions may induce environmental stress or compromise insect productivity and welfare. Escape behaviour, larval aggregation behaviour, possibly cannibalism in the larval stage and mating frequency and reproduction rate are identified as behaviours providing information on welfare of larval and adult flies. Finally, a number of aspects are identified for which behavioural knowledge is currently still sparse, while this may be important to safeguard insect welfare. We conclude with recommendations for future research to promote insect welfare.

Entomological identification of the post-mortem colonization of wolf cadavers in different decomposition stages

Human-driven biodiversity loss is progressively becoming a problem with dramatic consequences for the conservation of vital ecosystems. The increasing number of illegal killings of the grey wolf (Canis lupus, Linnaeus, 1758), a threatened species, displays the need for investigation and prosecution of such offences. Forensic entomology makes use of the knowledge about necrophagous insects to estimate a minimum time-since-death interval of the deceased person or animal, which can give important information on a possible perpetrator. The cadaver fauna along five decomposition stages of wolves in Germany was investigated in the period 2014–2021. The insects from 70 wolf cadavers, originating from all over Germany, were provided by the Leibniz Institute for Zoo and Wildlife Research Berlin. The accumulated degree day (ADD) model was applied for the post-mortem interval estimation on wolf cadavers for the first time. A total of 20 coleopteran species and 14 different dipteran species were discovered and identified. Almost 99 % of all insect specimens were from the order of Diptera, and beetles (Coleoptera) accounted for only 1 % of the cadaver fauna. The blowflies (Calliphoridae) are of particular importance for forensic issues, accounting for about 66 % of all families. Carrion beetles (Silphidae) were found as the second most abundant family (about 21 %). In addition, combining all cases, a steadily increasing insect species richness S was detected from early decay to advanced decay (fresh S = 8; bloated S = 12; active decay S = 21; advanced decay S = 34). In the following remains stage, the species number decreased again (S = 24). However, no significant difference in the number of species was found between the stages of decay when the cases were considered individually. The temporal pattern of insect appearance was found to be congruent with those of previous studies. Furthermore, a time of death was determined for each case and compared to the pathologist's estimates. This study provides insights into the arthropod fauna of wolf remains for the first time, applies the ADD-Model for post-mortem interval estimation, and discusses the suitability of forensic entomology for wildlife death investigations.

The interaction between the Dbf4 ortholog Chiffon and Gcn5 is conserved in Dipteran insect species.

Chiffon is the sole Drosophila ortholog of Dbf4, the regulatory subunit for the cell-cycle kinase Cdc7 that initiates DNA replication. In Drosophila, the chiffon gene encodes two polypeptides with independent activities. Chiffon-A contains the conserved Dbf4 motifs and interacts with Cdc7 to form the Dbf4-dependent Kinase (DDK) complex, which is essential for a specialized form of DNA replication. In contrast, Chiffon-B binds the histone acetyltransferase Gcn5 to form the Chiffon histone acetyltransferase (CHAT) complex, which is necessary for histone H3 acetylation and viability. Previous studies have shown that the Chiffon-B region is only present within insects. However, it was unclear how widely the interaction between Chiffon-B and Gcn5 was conserved among insect species. To examine this, we performed yeast two-hybrid assays using Chiffon-B and Gcn5 from a variety of insect species and found that Chiffon-B and Gcn5 interact in Diptera species such as Australian sheep blowfly and yellow fever mosquito. Protein domain analysis identified that Chiffon-B has features of acidic transcriptional activators such as Gal4 or VP16. We propose that the CHAT complex plays a critical role in a biological process that is unique to Dipterans and could therefore be a potential target for pest control strategies.

Autofluorescent Biomolecules in Diptera: From Structure to Metabolism and Behavior.

Light-based phenomena in insects have long attracted researchers’ attention. Surface color distribution patterns are commonly used for taxonomical purposes, while optically-active structures from Coleoptera cuticle or Lepidoptera wings have inspired technological applications, such as biosensors and energy accumulation devices. In Diptera, besides optically-based phenomena, biomolecules able to fluoresce can act as markers of bio-metabolic, structural and behavioral features. Resilin or chitinous compounds, with their respective blue or green-to-red autofluorescence (AF), are commonly related to biomechanical and structural properties, helpful to clarify the mechanisms underlying substrate adhesion of ectoparasites’ leg appendages, or the antennal abilities in tuning sound detection. Metarhodopsin, a red fluorescing photoproduct of rhodopsin, allows to investigate visual mechanisms, whereas NAD(P)H and flavins, commonly relatable to energy metabolism, favor the investigation of sperm vitality. Lipofuscins are AF biomarkers of aging, as well as pteridines, which, similarly to kynurenines, are also exploited in metabolic investigations. Beside the knowledge available in Drosophila melanogaster, a widely used model to study also human disorder and disease mechanisms, here we review optically-based studies in other dipteran species, including mosquitoes and fruit flies, discussing future perspectives for targeted studies with various practical applications, including pest and vector control.

Pollination efficiency and foraging behaviour of honey bees and flies to onion Allium cepa L.

Bees and flies are the most common visitors to onion flowers in Karnataka, but evidence for their pollination efficiency is lacking. Hence, experiments on pollination efficiency and behavioral observations were recorded using four bees (Apidae: Hymenoptera) and two flies (Syrphidae: Diptera) species visiting onion flowers (Allium cepa L.) at the experimental fields of ICAR-Indian Institute of Horticultural Research, Bengaluru, Karnataka, India (13.13°N; 77.49°E) during 2020-2021. Pollinator effectiveness (PE) is measured as pollen deposition on stigmas per visit. Honey bees and flies mainly collect nectar and stingless bees mainly pollen (81.90%), bringing the latter into intimate contact with the stigma more frequently. Stingless bees, honey bees and flies were often seen foraging together on the same umbel, and no antagonistic interactions were observed. The rock bee (Apis dorsata) deposited a maximum number (34.60) of pollen on stigma per visit followed by the stingless bee (24.45). The flowers pollinated by bees had a higher seed set percentage; the weight of 1000 seeds and the average number of seeds/umbels were higher compared to flowers deprived of pollinators. The results allowed us to conclude that bees are the effective pollinators of onions. Further work is needed to determine the optimum bee population required for maximizing the seed yield in onion.

Species Diversity, Habitat Distribution, and Blood Meal Analysis of Haematophagous Dipterans Collected by CDC-UV Light Traps in the Dominican Republic.

Haematophagous insects cause major economic losses by both direct damage and the transmission of pathogens. However, the biting Diptera species in the Caribbean region have been poorly documented. During 2021, CDC downdraft suction traps with UV light were employed to assess both the species occurrence and blood meal sources across three different habitats in the Dominican Republic. Eighteen species of mosquitoes (n = 274), six species of Culicoides (n = 803), two black fly species (n = 2), and one species of muscid fly (n = 25) were identified at species-level by morphology and/or molecular phylogenetic approaches based on the mitochondrial cytochrome c oxidase subunit 1 (COI). Engorged mosquito (n = 5) and Culicoides (n = 28) females showed host preferences derived exclusively from mammals (cows and pigs), except Culex species containing the blood of chickens. Our study provides new records of the Diptera Dominican catalogue (Culex salinarius for the Greater Antilles, Culicoides jamaicensis for Hispaniola, and Culicoides haitiensis and Culicoides borinqueni for the Dominican Republic), the first available COI DNA sequences of different Diptera in the GenBank, some pictures of diagnostic features of closely related specimens, spatial distribution across the habitats studied, and new insights on their feeding preferences in the Caribbean region.

Meeting a threat of the Anthropocene: Taste avoidance of metal ions by Drosophila

The Anthropocene Epoch poses a critical challenge for organisms: they must cope with new threats at a rapid rate. These threats include toxic chemical compounds released into the environment by human activities. Here, we examine elevated concentrations of heavy metal ions as an example of anthropogenic stressors. We find that the fruit fly Drosophila avoids nine metal ions when present at elevated concentrations that the flies experienced rarely, if ever, until the Anthropocene. We characterize the avoidance of feeding and egg laying on metal ions, and we identify receptors, neurons, and taste organs that contribute to this avoidance. Different subsets of taste receptors, including members of both Ir (Ionotropic receptor) and Gr (Gustatory receptor) families contribute to the avoidance of different metal ions. We find that metal ions activate certain bitter-sensing neurons and inhibit sugar-sensing neurons. Some behavioral responses are mediated largely through neurons of the pharynx. Feeding avoidance remains stable over 10 generations of exposure to copper and zinc ions. Some responses to metal ions are conserved across diverse dipteran species, including the mosquito Aedes albopictus. Our results suggest mechanisms that may be essential to insects as they face challenges from environmental changes in the Anthropocene.

Life Cycle and Biometric Study of Hydrotaea capensis (Wiedemann, 1818) (Diptera, Muscidae), a Species of Forensic Interest

Simple SummaryFrom the point of view of forensic entomology, the study of the development times of the species and the factors conditioning them are issues of great importance to estimate the minPMI. Hydrotaea capensis is a Muscidae (Diptera) of forensic interest because of its colonization preferences. Thus, there is a need to have more precise data about its biology to gain insights into the interpretation of its presence in corpses. In this study, the H. capensis life cycle was studied at four constant temperatures, 18, 20, 25 and 30 °C, by recording the duration of its different developmental stages and the length reached in each larval stage, as well as some biometric characteristics of the emerged adults. Our work is the first to provide data on the duration of the life cycle of this species and pointed to some alar features as relevant biometric adult indicators to be considered.One of the most important and perhaps most used applications of forensic entomology concerns the estimation of the minimum post-mortem interval (minPMI), defined as the time interval between death and the actual finding of a corpse. Some Diptera species are critical in these studies because they are the first ones capable of detecting and finding a corpse and are selectively attracted by its decomposing status. Thus, the knowledge of the micromorphology of their preimaginal stages and of their life cycles within a time frame constitutes solid indicators for estimating the minPMI. Hydrotaea capensis is a Muscidae of forensic interest usually considered as a late colonizer of corpses. It is widely distributed, living mainly in warm regions, and present in a wide variety of habitats. In this study, the H. capensis life cycle was studied at four constant temperatures, 18°, 20°, 25° and 30 °C, by recording the duration of its different developmental stages, including the length reached in each larval stage, as well as some biometric characteristics of the emerged adults. Significant differences were observed in the average time of development of most larval stages, with a longer duration at low temperatures, and in the length of each larval stage depending on the temperature, but, in this case, without a clear pattern. Moreover, significant differences were found in some alar features, pointing to them as a relevant indicator to be considered. The data provided will assist forensic entomologists to make more accurate minPMI estimations in cases where H. capensis is present.

Characterization of anti-microbial peptides and proteins from maggots of Calliphoridae and Sarcophagidae fly species (Diptera)

Removal of infected wounds using maggots has been known for centuries. Early research has shown that the maggot exosecretion, whole body, and fecal waste products of Calliphoridae and Sarcophagidae species contain a variety of alkaline peptides capable of inhibiting bacterial growth. Since the wide application of antibiotics such as penicillin, a number of bacterial infections have become insensitive to antibiotic treatment. In many of these instances, maggot therapy has been successfully applied for the treatment of chronic wounds. To identify and compare the expression patterns of anti-microbial peptides (AMPs) from some dipteran species, transcriptome analyses were conducted for the maggots of 11 Calliphoridae and Sarcophagidae species. Species of the subfamily Calliphorinae showed relatively higher expression levels of AMPs and anti-microbial proteins compared with those of Luciliinae and Sarcophagidae species. Furthermore, among all of the dipteran species examined, Lucilia illustris exhibited the highest transcription levels of AMPs. Cecropin A2 and defensin, whose expression levels were the highest among the anti-microbial peptides, were synthesized to test their biological activity. The synthesized peptides showed anti-microbial activities without hemolytic activities. In particular, cecropin A2 of L. illustris exhibited the highest anti-microbial activity against all of the bacteria and fungi examined, thereby possessing the potential to be developed as a new alternative to antibiotics. This comparative transcriptomic study may provide new insights into anti-microbial compositions of some dipteran species.

Rearrangements in the musculature correlate with jumping behaviour in legless Mediterranean fruit fly larvae Ceratitis capitata (Tephritidae)

Larvae of holometabolic insects evolved different crawling strategies depending on the presence or absence of larval legs or life style. A rather unusual mode of locomotion has independently evolved in legless larvae of several dipteran species. Maggots of the Mediterranean fruit fly Ceratitis capitata developed an effective jumping mechanism to increase locomotion speed or to deter predators during the search for suitable pupation sites. Here, we use high-speed videography to visualize even the fastest movements during jump preparation and take-off. Quantification of kinetic and biometric parameters reveal that maggots jump up to 15-fold of their body length from a standing position and gain speed with 27 times the acceleration of gravity. Videos at high spatial resolution show the mechanism of latch formation and release in unprecedented detail. Mouth hooks insert in the caudal segment and raise a cuticular fold that serves as a handle to pressurize the body prior to launch. Since locomotion behaviour should be intrinsically linked to neuromuscular systems, we dissected third instar larvae and determined the precise pattern of abdominal muscles fibres. Compared to non-jumping dipteran larvae, such as Drosophila melanogaster, the overall arrangement is highly similar, but a few muscle fibres show characteristic re-arrangements in orientation and strength that are consistent with a role in bending and jumping. These results suggest that body wall muscles show adaptations to jumping behaviour in Ceratitis larvae, and possibly also in other species with different jumping techniques.

Possible functions of the petal hairs of Jodina Hook. & Arn. ex Meisn. (Cervantesiaceae) during pollination

Jodina (Cervantesiaceae, Santalales) is a South American monotypic genus of hemiparasitic trees that occurs mainly in xeric and semi-xeric habitats. The genus is characterized by its rhombic, glabrous and stiff leaves. The flowers are monochlamydous and the petals give rise to long hairs arranged behind the stamens. It has been mentioned that these hairs could play a role during pollination nevertheless no information is recorded for Jodina or other members of Cervantesiaceae. The goals of this study were: (1) to deepen the anatomical analyses of the petal hairs and the disc, including chemistry; (2) to ascertain the possible role of the petal hairs during pollination. Flowers in different stages of development were collected in natural populations of Argentina and processed for LM and SEM analyses. Additionally, an olfactory test was conducted. We observed that, as the flower opens, the hairs enter the thecae and emerge with pollen grains adhered at their tips. The petal hairs contain proteins and lipids. They also stain light red with neutral red and emit a delicate pleasant fragrance. During field trips two species of Diptera were observed visiting the flowers. The findings let us to suggest a possible role of the petal hairs as osmophores to attract insects and as a reward for pollinators.

Catalogue of the Diptera (Insecta) of Morocco- an annotated checklist, with distributions and a bibliography.

The faunistic knowledge of the Diptera of Morocco recorded from 1787 to 2021 is summarized and updated in this first catalogue of Moroccan Diptera species. A total of 3057 species, classified into 948 genera and 93 families (21 Nematocera and 72 Brachycera), are listed. Taxa (superfamily, family, genus and species) have been updated according to current interpretations, based on reviews in the literature, the expertise of authors and contributors, and recently conducted fieldwork. Data to compile this catalogue were primarily gathered from the literature. In total, 1225 references were consulted and some information was also obtained from online databases. Each family was reviewed and the checklist updated by the respective taxon expert(s), including the number of species that can be expected for that family in Morocco. For each valid species, synonyms known to have been used for published records from Morocco are listed under the currently accepted name. Where available, distribution within Morocco is also included. One new combination is proposed: Assuaniamelanoleuca (Séguy, 1941), comb. nov. (Chloropidae).

Histochemical staining techniques in Culex pipiens and Drosophila melanogaster (Diptera) with a comparison to mammals.

Insects play an important role in ecosystems. Changes in their abundance and biodiversity are of paramount interest, as there has not only been an alarming decline of insects important for ecosystem health throughout the past decades, but also an increase in insects detrimental for biomes. Furthermore, insects pose a threat to modern society as arbovirus-transmitting vectors. Therefore, detailed knowledge of insect staining characteristics could be beneficial as a basis for further studies, whether in the context of species conservation or control of insect pests. Thus, this study compared 14 histochemical stains for their usefulness in insects regarding nervous tissue, connective tissue components, mucins and polysaccharides, mineralization, and microorganisms. The study used formalin-fixed paraffin-embedded tissue sections of mammals (Equus caballus) and 2 dipterans (Culex pipiens biotype molestus, Drosophila melanogaster). Several histochemical stains were suitable for tissue assessment in insects and mammals, in particular for nervous tissue (Bielschowsky silver stain, luxol fast blue-cresyl violet) and polysaccharides (alcian blue, periodic acid-Schiff with and without diastase treatment, toluidine blue). Other stains proved useful for visualization of insect-specific organ characteristics such as Gomori's reticulin stain for tracheoles in both dipteran species, Heidenhain's azan for midgut-associated connective tissue, and von Kossa for mineral deposition in Malpighian tubules of C. pipiens biotype molestus. In summary, this study provides comparable insights into histochemical procedures in mammals and insects and their usefulness for histological assessment of C. pipiens biotype molestus and D. melanogaster.

Detection of F1 Hybrids from Single-genome Data Reveals Frequent Hybridization in Hymenoptera and Particularly Ants.

Hybridization occupies a central role in many fundamental evolutionary processes, such as speciation or adaptation. Yet, despite its pivotal importance in evolution, little is known about the actual prevalence and distribution of current hybridization across the tree of life. Here we develop and implement a new statistical method enabling the detection of F1 hybrids from single-individual genome sequencing data. Using simulations and sequencing data from known hybrid systems, we first demonstrate the specificity of the method, and identify its statistical limits. Next, we showcase the method by applying it to available sequencing data from more than 1,500 species of Arthropods, including Hymenoptera, Hemiptera, Coleoptera, Diptera, and Archnida. Among these taxa, we find Hymenoptera, and especially ants, to display the highest number of candidate F1 hybrids, suggesting higher rates of recent hybridization between previously isolated gene pools in these groups. The prevalence of F1 hybrids was heterogeneously distributed across ants, with taxa including many candidates tending to harbor specific ecological and life-history traits. This work shows how large-scale genomic comparative studies of recent hybridization can be implemented, uncovering the determinants of first-generation hybridization across whole taxa.

Preliminary Study on the Distribution and Diversity of Diptera at Tuba Island Reserve Forest, Langkawi Malaysia

Abstract Insects, the ecologically important organisms, contribute most to the world’s biodiversity. A study on distribution to diversity of order Diptera was conducted in a selected area of Tuba Island Forest Reserve, Langkawi from 17th September to 21st September 2020 using Malaise traps. Three study sites which were forest fringe, middle forest and inner forest were chosen. A total of 5450 individuals of Diptera belonging to 18 families and 27 morphospecies were collected in this study. Diptera samples were highly abundant in the forest fringe with Mycetophilidae as the most abundant family recorded, while the least abundant family was Drosophilidae. The Shannon-Weiner Diversity Index (H’) showed that the forest fringe had the highest diversity value (H’=2.05), followed by the inner forest (H’=0.67) and middle forest (H’=0.45). The highest Evenness Index (E’) value was recorded from the inner forest with E’=0.66 while the highest value for Margalef Richness Index (R’) was from the forest fringe which was 1.91. The Kruskal-Wallis H test conducted indicates that there was a significant difference in the distribution of Dipteran across the three locations of the forest with P<0.05. Overall, this study suggested that the diversity of order Diptera was higher in the forest fringe compared to the middle and the inner forest. This study is important as it is helpful for future study of Diptera species in Tuba Island as well as for conservation measures in the island ecosystem.