Search

Moose (Alces alces) in boreal habitats feed and rest where they are exposed to Dipteran flies and the parasites they carry. We collected 31,905 flies during the summer from 12 habituated moose on the Kenai Peninsula, Alaska. Moose flies, Haematobosca alcis (Snow), Diptera: Muscidae—a species that completes its entire life cycle on or around moose—accounted for 91% of flies collected; the reminder of the flies collected included mosquitoes (Culicidae), black flies (Simuliidae), and deer flies (Tabanidae). Flies impose physiological costs for moose, e.g. vectors for parasites such as Legworm (Onchocerca spp.) which causes sores on the hind legs of moose. We found that the number of sores present on the hind legs of moose is positively correlated with body fat, which suggests a correlation between gains of energy and damage from flies. We also found that the number of sores is negatively correlated with serum albumin, which is indicative of an inflammatory response and body protein being used to repair injuries from flies and parasites. The number or type of flies present on a Moose were not correlated with the concentration of corticosteroids in saliva or feces. Flies do not elicit a stress response in moose even though the costs of repairing wounds and resisting infections of those wounds likely reduce gains of protein from summer foraging. Moose can tolerate the injuries from biting flies with regular gains from summer foraging but exposure to insect-borne parasites poses a risk to reproduction and survival. [ABSTRACT FROM AUTHOR]

Numerous factors have been implicated in the cell–cell interactions that lead to elimination of cells via cell competition, a context-dependent process of cell selection in somatic tissues that is based on comparisons of cellular fitness. Here, we use a series of genetic tests in Drosophila to explore the relative contribution of the pleiotropic cytokine tumor necrosis factor α (TNFα) in Myc-mediated cell competition (also known as Myc supercompetition or Myc cell competition). We find that the sole Drosophila TNF, Eiger (Egr), its receptor Grindelwald (Grnd/TNF receptor), and the adaptor proteins Traf4 and Traf6 are required to eliminate wild-type "loser" cells during Myc cell competition. Although typically the interaction between Egr and Grnd leads to cell death by activating the intracellular Jun N-terminal kinase (JNK) stress signaling pathway, our experiments reveal that many components of canonical JNK signaling are dispensable for cell death in Myc cell competition, including the JNKKK Tak1, the JNKK Hemipterous and the JNK Basket. Our results suggest that Egr/Grnd signaling participates in Myc cell competition but functions in a role that is largely independent of the JNK signaling pathway. [ABSTRACT FROM AUTHOR]

An attractive perfume is a complex mixture of compounds, some of which may be unpleasant on their own. This is also true for the volatile combinations from yeast fermentation products in vineyards and orchards when assessed by Drosophila. Here, we used crosses between a yeast strain with an attractive fermentation profile and another strain with a repulsive one and tested fly responses using a T-maze. QTL analysis reveals allelic variation in four yeast genes, namely PTC6 , SAT4 , YFL040W , and ARI1 , that modulated expression levels of volatile compounds [assessed by gas chromatography–mass spectrometry (GC–MS)] and in different combinations, generated various levels of attractiveness. The parent strain that is more attractive to Drosophila has repulsive alleles at two of the loci, while the least attractive parent has attractive alleles. Behavioral assays using artificial mixtures mimicking the composition of odors from fermentation validated the results of GC–MS and QTL mapping, thereby directly connecting genetic variation in yeast to attractiveness in flies. This study can be used as a basis for dissecting the combination of olfactory receptors that mediate the attractiveness/repulsion of flies to yeast volatiles and may also serve as a model for testing the attractiveness of pest species such as Drosophila suzukii to their host fruit. [ABSTRACT FROM AUTHOR]

Chitosan is a naturally derived polymer that has significant potential for use as a bioinsecticide. Despite this, there is a lack of research as to the efficacy of chitosan for many insect pest species. The apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae), is one such pest for which chitosan toxicity has not been explored. In this study, the toxicity of chitosan for R. pomonella adults was tested via no-choice feeding assays. An aging trial was further used to test the mortality of flies provided dried chitosan-sucrose treatments (CST), which were aged for 0 or 3 days in greenhouse conditions. This study found that the CST is toxic for R. pomonella adults when ingested, leading to a significant increase in the rate of mortality compared to control groups. The use of dried chitosan, however, did not change the mortality of flies, suggesting this will not be an effective delivery mechanism. Effective biopesticide delivery systems have not been defined for chitosan use outside of a laboratory, indicating the need for further research testing delivery mechanisms. It is suggested that an effective method of delivery can be as a food-based bait in attract-and-kill traps, as chitosan must be ingested to kill flies. Forming a viscous solution, chitosan may be best suited to these lure systems. [ABSTRACT FROM AUTHOR]

Macronutrient intake impacts physiology, behavior, and gene expression in a wide range of organisms. We used the response surface methodology to compare how life history traits, lifespan, and reproduction differ as a function of protein and carbohydrate intakes under choice and no-choice feeding regimens in the fruit fly, Drosophila melanogaster. We found that when offered a choice of nutritionally complementary foods mated female flies regulated toward a protein to carbohydrate ratio (P:C) that was associated with shortened lifespan and maximal egg production when compared to response surfaces derived from flies fed 1 of a range of fixed diets differing in P:C (no-choice regimen). This difference in lifespan between choice and no-choice feeding was not seen in males or virgin flies, reflecting the fact that increased protein intake is triggered by mating to support egg production. However, whereas in mated females a higher P:C intake was associated with greater egg production under both choice and no-choice feeding, contrary to expectations, choice-fed mated flies laid fewer eggs than no-choice flies on equivalent macronutrient intakes, perhaps reflecting that they had to ingest twice the volume of food to attain an equivalent intake of nutrients than no-choice flies on a diet of equivalent P:C ratio. [ABSTRACT FROM AUTHOR]

The maternal inheritance of mitochondria is a widely accepted paradigm, and mechanisms that prevent paternal mitochondria transmission to offspring during spermatogenesis and postfertilization have been described. Although certain species do retain paternal mitochondria, the factors affecting paternal mitochondria inheritance in these cases are unclear. More importantly, the evolutionary benefit of retaining paternal mitochondria and their ultimate fate are unknown. Here we show that transplanted exogenous paternal D. yakuba mitochondria can be transmitted to offspring when maternal mitochondria are dysfunctional in D. melanogaster. Furthermore, we show that the preserved paternal mitochondria are functional, and can be stably inherited, such that the proportion of paternal mitochondria increases gradually in subsequent generations. Our work has important implications that paternal mitochondria inheritance should not be overlooked as a genetic phenomenon in evolution, especially when paternal mitochondria are of significant differences from the maternal mitochondria or the maternal mitochondria are functionally abnormal. Our results improve the understanding of mitochondrial inheritance and provide a new model system for its study. [ABSTRACT FROM AUTHOR]

The level of resistance to radiation and the developmental and molecular responses can vary between species, and even between developmental stages of one species. For flies (order: Diptera), prior studies concluded that the fungus gnat Bradysia (Sciara) coprophila (sub-order: Nematocera) is more resistant to irradiation-induced mutations that cause visible phenotypes than the fruit fly Drosophila melanogaster (sub-order: Brachycera). Therefore, we characterized the effects of and level of resistance to ionizing radiation on B. coprophila throughout its life cycle. Our data show that B. coprophila embryos are highly sensitive to even low doses of gamma-irradiation, whereas late-stage larvae can tolerate up to 80 Gy (compared to 40 Gy for D. melanogaster) and still retain their ability to develop to adulthood, though with a developmental delay. To survey the genes involved in the early transcriptional response to irradiation of B. coprophila larvae, we compared larval RNA-seq profiles with and without radiation treatment. The up-regulated genes were enriched for DNA damage response genes, including those involved in DNA repair, cell cycle arrest, and apoptosis, whereas the down-regulated genes were enriched for developmental regulators, consistent with the developmental delay of irradiated larvae. Interestingly, members of the PARP and AGO families were highly up-regulated in the B. coprophila radiation response. We compared the transcriptome responses in B. coprophila to the transcriptome responses in D. melanogaster from 3 previous studies: whereas pathway responses are highly conserved, specific gene responses are less so. Our study lays the groundwork for future work on the radiation responses in Diptera. [ABSTRACT FROM AUTHOR]

Larval habitats of blood-feeding stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), overlap with foraging sites of black blow flies, Phormia regina (Meigen) (Diptera: Calliphoridae). We tested the hypothesis that bacteria in blow fly excreta inform oviposition decisions by female stable flies. In laboratory 2-choice bioassays, we offered gravid female stable flies fabric-covered agar plates as oviposition sites that were kept sterile or inoculated with either a blend of 7 bacterial strains isolated from blow fly excreta (7-isolate-blend) or individual bacterial isolates from that blend. The 7-isolate-blend deterred oviposition by female stable flies, as did either of 2 strains of Morganella morganii subsp. sibonii. Conversely, Exiguobacterium sp. and Serratia marcescens each prompted oviposition by flies. The flies' oviposition decisions appear to be guided by bacteria-derived semiochemicals as the bacteria could not be physically accessed. Oviposition deterrence caused by semiochemicals of the 7-isolate-blend may help stable flies avoid competition with blow flies. The semiochemicals of bioactive bacterial strains could be developed as trap lures to attract and capture flies and deter their oviposition in select larval habitats. [ABSTRACT FROM AUTHOR]

Methylphosphate Capping Enzyme (MePCE) monomethylates the gamma phosphate at the 5′ end of the 7SK noncoding RNA, a modification thought to protect 7SK from degradation. 7SK serves as a scaffold for assembly of a snRNP complex that inhibits transcription by sequestering the positive elongation factor P-TEFb. While much is known about the biochemical activity of MePCE in vitro, little is known about its functions in vivo, or what roles—if any—there are for regions outside the conserved methyltransferase domain. Here, we investigated the role of Bin3, the Drosophila ortholog of MePCE, and its conserved functional domains in Drosophila development. We found that bin3 mutant females had strongly reduced rates of egg-laying, which was rescued by genetic reduction of P-TEFb activity, suggesting that Bin3 promotes fecundity by repressing P-TEFb. bin3 mutants also exhibited neuromuscular defects, analogous to a patient with MePCE haploinsufficiency. These defects were also rescued by genetic reduction of P-TEFb activity, suggesting that Bin3 and MePCE have conserved roles in promoting neuromuscular function by repressing P-TEFb. Unexpectedly, we found that a Bin3 catalytic mutant (Bin3Y795A) could still bind and stabilize 7SK and rescue all bin3 mutant phenotypes, indicating that Bin3 catalytic activity is dispensable for 7SK stability and snRNP function in vivo. Finally, we identified a metazoan-specific motif (MSM) outside of the methyltransferase domain and generated mutant flies lacking this motif (Bin3ΔMSM). Bin3ΔMSM mutant flies exhibited some—but not all— bin3 mutant phenotypes, suggesting that the MSM is required for a 7SK-independent, tissue-specific function of Bin3. [ABSTRACT FROM AUTHOR]

Epithelial cells contain polarity complexes on the lateral membrane and are organized in a hexagon-dominated polygonal array. The mechanisms regulating the organization of polygonal architecture in metazoan embryogenesis are not completely understood. Drosophila embryogenesis enables mechanistic analysis of epithelial polarity formation and its impact on polygonal organization. The plasma membrane (PM) of syncytial Drosophila blastoderm embryos is organized as a polygonal array with pseudocleavage furrow formation during the almost synchronous cortical division cycles. We find that polygonal (PM) organization arises in the metaphase (MP) of division cycle 11, and hexagon dominance occurs with an increase in furrow length in the metaphase of cycle 12. There is a decrease in cell shape index in metaphase from cycles 11 to 13. This coincides with Drosophila E-cad (DE-cadherin) and Bazooka enrichment at the edges and the septin, Peanut at the vertices of the furrow. We further assess the role of polarity and adhesion proteins in pseudocleavage furrow formation and its organization as a polygonal array. We find that DE-cadherin depletion leads to decreased furrow length, loss of hexagon dominance, and increased cell shape index. Bazooka and Peanut depletion lead to decreased furrow length, delay in onset of hexagon dominance from cycle 12 to 13, and increased cell shape index. Hexagon dominance occurs with an increase in furrow length in cycle 13 and increased DE-cadherin, possibly due to the inhibition of endocytosis. We conclude that polarity protein recruitment and regulation of endocytic pathways enable pseudocleavage furrow stability and the formation of a hexagon-dominated polygon array. [ABSTRACT FROM AUTHOR]

Large genome structural variations can impact genome regulation and integrity. Repeat-rich regions like pericentric heterochromatin are vulnerable to structural rearrangements although we know little about how often these rearrangements occur over evolutionary time. Repetitive genome regions are particularly difficult to study with genomic approaches, as they are missing from most genome assemblies. However, cytogenetic approaches offer a direct way to detect large rearrangements involving pericentric heterochromatin. Here, we use a cytogenetic approach to reveal large structural rearrangements associated with the X pericentromeric region of Drosophila simulans. These rearrangements involve large blocks of satellite DNA—the 500-bp and Rsp-like satellites—which colocalize in the X pericentromeric heterochromatin. We find that this region is polymorphic not only among different strains, but between isolates of the same strain from different labs, and even within individual isolates. On the one hand, our observations raise questions regarding the potential impact of such variation at the phenotypic level and our ability to control for such genetic variability. On the other hand, this highlights the very rapid turnover of the pericentric heterochromatin most likely associated with genomic instability of the X pericentromere. It represents a unique opportunity to study the dynamics of pericentric heterochromatin, the evolution of associated satellites on a very short time scale, and to better understand how structural variation arises. [ABSTRACT FROM AUTHOR]

Aims We investigate extraintestinal pathogenic genes (ExPEC) related to virulence of Escherichia coli in flies from the dairy environment. Methods and Results We collected 217 flies from nine dairy farms, which were submitted to microbiological culture. Fifty-one E. coli were identified using mass spectrometry. Eleven dipteran families were identified, with a predominance of Muscidae, and a minor frequency of Tachinidae, Drosophilidae, Sphaeroceridae, Ulidiidae, Syrphidae, Chloropidae, Calliphoridae, Sarcophagidae, and Piophilidae. A panel of 16 virulence-encoding genes related to ExPEC infections were investigated, which revealed predominance of serum resistance (traT , 31/51 = 60.8%; ompT , 29/51 = 56.9%), iron uptake (irp2 , 17/51 = 33.3%, iucD 11/51 = 21.6%), and adhesins (papC , 6/51 = 11.8%; papA , 5/51 = 9.8%). Conclusions Our findings reveal Dipterans from milking environment carrying ExPEC virulence-encoding genes also identified in clinical bovine E. coli -induced infections. [ABSTRACT FROM AUTHOR]

Autophagy, an autophagosome and lysosome-based eukaryotic cellular degradation system, has previously been implicated in lifespan regulation in different animal models. In this report, we show that expression of the RNAi transgenes targeting the transcripts of the key autophagy genes Atg1 or Atg18 in adult fly muscle or glia does not affect the overall levels of autophagosomes in those tissues and does not change the lifespan of the tested flies but the lifespan reduction phenotype has become apparent when Atg1 RNAi or Atg18 RNAi is expressed ubiquitously in adult flies or after autophagy is eradicated through the knockdown of Atg1 or Atg18 in adult fly adipocytes. Lifespan reduction was also observed when Atg1 or Atg18 was knocked down in adult fly enteroblasts and midgut stem cells. Overexpression of wild-type Atg1 in adult fly muscle or adipocytes reduces the lifespan and causes accumulation of high levels of ubiquitinated protein aggregates in muscles. Our research data have highlighted the important functions of the key autophagy genes in adult fly adipocytes, enteroblasts, and midgut stem cells and their undetermined roles in adult fly muscle and glia for lifespan regulation. [ABSTRACT FROM AUTHOR]

The "insects as food and feed" movement is gaining considerable momentum as a novel means to provide protein to people (i.e. food) and other animals (i.e. feed). Insects require significantly fewer resources, such as water and land, to produce, process, and distribute as a food or feed source. While the production of insect biomass has received considerable attention for use as food or feed, little is known about the value of the residual materials remaining after digestion. One insect, the black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae), can generate large quantities of residual (i.e. frass) that is high in nitrogen, phosphorus, and potassium. These materials could serve as a partial replacement for fertilizer or peat, thus creating added value to the insects as food and feed sector. Greenhouse studies were designed to investigate the use of frass in vegetable production. In pot studies with tomatoes, different ratios of peat:vermicompost and peat:insect frass were compared to a 100% peat control. Across all other parameters, tomato fruits and vegetative biomass did not produce significant differences across treatments, indicating results were comparable to the control (i.e. 100% peat). Thus, replacing peat with black soldier fly frass is a viable option and could allow for the peat industry to become more sustainable and regenerative. However, it should be noted that average individual tomato fruit weight was significantly (P  < 0.05) higher (by 19%) in the vermicompost 10% treatment compared to the control, which did not differ from treatments including black soldier fly frass. [ABSTRACT FROM AUTHOR]

In many arthropods, including insects responsible for transmission of human diseases, behaviors that include mating, aggregation, and aggression are triggered by detection of pheromones. Extracellular odorant binding proteins are critical for pheromone detection in many insects and are secreted into the fluid bathing the olfactory neuron dendrites. In Drosophila melanogaster, the odorant binding protein LUSH is essential for normal sensitivity to the volatile sex pheromone, 11-cis vaccenyl acetate (cVA). Using a genetic screen for cVA pheromone insensitivity, we identified ANCE-3, a homolog of human angiotensin converting enzyme that is required for detection of cVA pheromone. The mutants have normal dose-response curves for food odors, although olfactory neuron amplitudes are reduced in all olfactory neurons examined. ance-3 mutants have profound delays in mating, and the courtship defects are primarily but not exclusively due to loss of ance-3 function in males. We demonstrate that ANCE-3 is required in the sensillae support cells for normal reproductive behavior, and that localization of odorant binding proteins to the sensillum lymph is blocked in the mutants. Expression of an ance-3 cDNA in sensillae support cells completely rescues the cVA responses, LUSH localization, and courtship defects. We show the courtship latency defects are not due to effects on olfactory neurons in the antenna nor mediated through ORCO receptors, but instead stem from ANCE-3-dependent effects on chemosensory sensillae in other body parts. These findings reveal an unexpected factor critical for pheromone detection with profound influence on reproductive behaviors. [ABSTRACT FROM AUTHOR]

The chromatin of animal cells contains two types of histones: canonical histones that are expressed during S phase of the cell cycle to package the newly replicated genome, and variant histones with specialized functions that are expressed throughout the cell cycle and in nonproliferating cells. Determining whether and how canonical and variant histones cooperate to regulate genome function is integral to understanding how chromatin-based processes affect normal and pathological development. Here, we demonstrate that variant histone H3.3 is essential for Drosophila development only when canonical histone gene copy number is reduced, suggesting that coordination between canonical H3.2 and variant H3.3 expression is necessary to provide sufficient H3 protein for normal genome function. To identify genes that depend upon, or are involved in, this coordinate regulation we screened for heterozygous chromosome 3 deficiencies that impair development of flies bearing reduced H3.2 and H3.3 gene copy number. We identified two regions of chromosome 3 that conferred this phenotype, one of which contains the Polycomb gene, which is necessary for establishing domains of facultative chromatin that repress master regulator genes during development. We further found that reduction in Polycomb dosage decreases viability of animals with no H3.3 gene copies. Moreover, heterozygous Polycomb mutations result in de-repression of the Polycomb target gene Ubx and cause ectopic sex combs when either canonical or variant H3 gene copy number is reduced. We conclude that Polycomb-mediated facultative heterochromatin function is compromised when canonical and variant H3 gene copy number falls below a critical threshold. [ABSTRACT FROM AUTHOR]

Epigenetic regulation varies with the environment. In the fruit fly Drosophila melanogaster, environmental temperature can affect chromatin-based gene regulation. Genes regulated by the Polycomb group can vary in their transcriptional output in response to changes in temperature, which typically increases with decreasing temperature. Here, we studied temperature-sensitive expression of Polycomb group target genes on a genome-wide scale, as well as temperature-sensitive enrichment of two histone modifications associated with the regulation of Polycomb group target genes, H3K27me3 and H3K4me3. We investigated temperature-sensitivity in adult flies, and possible differences thereof between populations adapted to temperate and tropical climates. Compared to genes not targeted by the Polycomb group, an elevated number of target genes showed higher expression at the lower temperature, as it is typically observed for Polycomb group regulation. Many of the Polycomb group target genes also exhibited temperature-sensitive H3K4me3 enrichment in the same direction, and the H3K4me3 temperature response correlated positively with that of expression. A small set of target sites also showed temperature-sensitive enrichment of H3K27me3, again with a higher proportion corresponding to increased transcriptional activation at the lower temperature. Overall, higher transcriptional activity at lower temperature was less pronounced in males compared to females, and in temperate compared to tropical flies. Possible trans- and cis-acting factors responsible for reduced expression plasticity in temperate flies were identified, including factors belonging to the Trithorax group and insulator binding proteins, respectively. [ABSTRACT FROM AUTHOR]

Retrotransposons are a class of transposable elements capable of self-replication and insertion into new genomic locations. Across species, the mobilization of retrotransposons in somatic cells has been suggested to contribute to the cell and tissue functional decline that occurs during aging. Retrotransposons are broadly expressed across cell types, and de novo insertions have been observed to correlate with tumorigenesis. However, the extent to which new retrotransposon insertions occur during normal aging and their effect on cellular and animal function remains understudied. Here, we use a single nucleus whole genome sequencing approach in Drosophila to directly test whether transposon insertions increase with age in somatic cells. Analyses of nuclei from thoraces and indirect flight muscles using a newly developed pipeline, Retrofind, revealed no significant increase in the number of transposon insertions with age. Despite this, reducing the expression of two different retrotransposons, 412 and Roo, extended lifespan, but did not alter indicators of health such as stress resistance. This suggests a key role for transposon expression and not insertion in regulating longevity. Transcriptomic analyses revealed similar changes to gene expression in 412 and Roo knockdown flies and highlighted changes to genes involved in proteolysis and immune function as potential contributors to the observed changes in longevity. Combined, our data show a clear link between retrotransposon expression and aging. [ABSTRACT FROM AUTHOR]

Originally a genetic model organism, the experimental use of Drosophila melanogaster has grown to include quantitative behavioral analyses, sophisticated perturbations of neuronal function, and detailed sensory physiology. A highlight of these developments can be seen in the context of vision, where pioneering studies have uncovered fundamental and generalizable principles of sensory processing. Here we begin with an overview of vision-guided behaviors and common methods for probing visual circuits. We then outline the anatomy and physiology of brain regions involved in visual processing, beginning at the sensory periphery and ending with descending motor control. Areas of focus include contrast and motion detection in the optic lobe, circuits for visual feature selectivity, computations in support of spatial navigation, and contextual associative learning. Finally, we look to the future of fly visual neuroscience and discuss promising topics for further study. [ABSTRACT FROM AUTHOR]

Bactrocera tsuneonis and Bactrocera minax are the most destructive pests that damage citrus in China. These key pests hinder the citrus trade, cause significant financial losses, drastically lower citrus production and quality, and decrease farmer enthusiasm for citrus planting. Bactrocera minax and B. tsuneonis are very similar in all life stages. There are limited morphological characteristics to differentiate the adult species, and it is nearly impossible to differentiate these two species in the egg and larval stages. Loop-mediated isothermal amplification (LAMP) is a rapid and robust diagnostic tool used to identify these two species accurately. We designed two sets of primers to distinguish B. minax and B. tsuneonis using DNA barcoding region of the COI gene. Only 50 min was needed under a constant temperature of 65ºC to determine the species of the two flies. The reaction system has high specificity and sensitivity, in which these two species can be accurately distinguished between different geographical populations and 1.0 ng/μL was the lowest DNA concentration that could be detected. Our primers can quickly identify these key pests without knowing their morphology, which could facilitate plant protection workers at the primary level to solve problems in plant quarantine. [ABSTRACT FROM AUTHOR]

Sympatric host-associated genetic differentiation is a prominent pattern that could lead to speciation. In insects, there are numerous examples of host-associated differentiation among herbivores that prefer different plants, and parasitoids that prefer different hosts, but few examples for specialist predators. We developed new microsatellite loci for two species of silver fly, Leucotaraxis argenticollis (Zetterstedt) and L. piniperda (Malloch) (Diptera: Chamaemyiidae), being evaluated as biological control agents for the hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), in eastern North America where it is a nonnative pest. We obtained DNA from specimens of both fly species feeding on native A. tsugae in western North America, as well as on other western and eastern adelgid species. We performed population genetic analyses using the new loci and DNA barcode sequences. Our results confirmed east–west allopatric divergence and uncovered nested genetic differentiation associated with different adelgid prey species and their host plants in western North America for both species of silver flies. For both species, there is also evidence for a longer history of diversification in the west, with ancestral specialization of feeding on pine adelgids, which was retained after range expansion to the east. More recently, divergence to feeding on new adelgid prey species occurred in the west. Our findings are consistent with the hypothesis that host-alternating life cycles in Adelgidae may provide temporary escape from specialist predators. We discuss the implications for biological control efficacy and potential for lineage hybridization as western flies are released in the east to control A. tsugae. [ABSTRACT FROM AUTHOR]

Nematocera (lower Diptera) is a taxonomically diverse group with ~52 000 species in 40 families, including many important agricultural and sanitary pests. The number and composition of nematoceran infraorders have long been subject to debate. The use of mitochondrial genomes for resolving these questions shows considerable promise. Herein, we sequenced and annotated the complete or nearly complete mitochondrial genomes of eight nematoceran species from seven families, representing the first mitochondrial genomes of two infraorders and six families. They range in size from 14 447 to 16 255 bp and all contain 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes and an AT-rich region. Like other nematocerans, the arrangement of genes of their mitochondrial genomes is identical to the presumed ancestral arrangement. A phylogenetic analysis was conducted by using mitochondrial genomes of 116 representatives from Diptera, Mecoptera and Siphonaptera. Our phylogenetic results support both new and traditional arrangements. The small, highly specialized family Deuterophlebiidae appears to be sister to all remaining Diptera, while Nymphomyiidae and Blephariceridae are nested within Culicomorpha and Psychodomorpha, respectively; hence, the recognition of 'Blephariceromorpha' as a valid monophyletic infraorder is discouraged. Anisopodidae is supported as the sister group of Brachycera. [ABSTRACT FROM AUTHOR]

In mammals, energy homeostasis is regulated by the antagonistic action of hormones insulin and glucagon. However, in contrast to the highly conserved insulin, glucagon is absent in most invertebrates. Although there are several endocrine regulators of energy expenditure and catabolism (such as the adipokinetic hormone), no single invertebrate hormone with all of the functions of glucagon has been described so far. Here, we used genetic gain- and loss-of-function experiments to show that the Drosophila gene Ion transport peptide (ITP) codes for a novel catabolic regulator that increases energy expenditure, lowers fat and glycogen reserves, and increases glucose and trehalose. Intriguingly, Ion transport peptide has additional functions reminiscent of glucagon, such as inhibition of feeding and transit of the meal throughout the digestive tract. Furthermore, Ion transport peptide interacts with the well-known signaling via the Adipokinetic hormone; Ion transport peptide promotes the pathway by stimulating Adipokinetic hormone secretion and transcription of the receptor AkhR. The genetic manipulations of Ion transport peptide on standard and Adipokinetic hormone-deficient backgrounds showed that the Adipokinetic hormone peptide mediates the hyperglycemic and hypertrehalosemic effects of Ion transport peptide, while the other metabolic functions of Ion transport peptide seem to be Adipokinetic hormone independent. In addition, Ion transport peptide is necessary for critical processes such as development, starvation-induced foraging, reproduction, and average lifespan. Altogether, our work describes a novel master regulator of fly physiology with functions closely resembling mammalian glucagon. [ABSTRACT FROM AUTHOR]

Sex‐based differences in physiological traits may be influenced by both evolutionary and environmental factors. Here we used male and female flies from >80 Drosophila species reared under common conditions to examine variance in a number of physiological traits including size, starvation, desiccation and thermal tolerance. Sex‐based differences for desiccation and starvation resistance were comparable in magnitude to those for size, with females tending to be relatively more resistant than males. In contrast thermal resistance showed low divergence between the sexes. Phylogenetic signal was detected for measures of divergence between the sexes, such that species from the Sophophora clade showed larger differences between the sexes than species from the Drosophila clade. We also found that sex‐based differences in desiccation resistance, body size and starvation resistance were weakly associated with climate (annual mean temperature/precipitation seasonality) but the direction and association with environment depended on phylogenetic position. The results suggest that divergence between the sexes can be linked to environmental factors, while an association with phylogeny suggests sex‐based differences persist over long evolutionary time‐frames. [ABSTRACT FROM AUTHOR]

Structurally colored sexual signals are a conspicuous and widespread class of ornament used in mate choice, though the extent to which they encode information on the quality of their bearers is not fully resolved. Theory predicts that signaling traits under strong sexual selection as honest indicators should evolve to be more developmentally integrated and exaggerated than nonsexual traits, thereby leading to heightened condition dependence. Here, we test this prediction through examination of the sexually dimorphic faces and wings of the cursorial fly Lispe cana. Males and females possess structural UV-white and golden faces, respectively, and males present their faces and wings to females during close-range, ground-based courtship displays, thereby creating the opportunity for mutual inspection. Across a field-collected sample of individuals, we found that the appearance of the faces of both sexes scaled positively with individual condition, though along separate axes. Males in better condition expressed brighter faces as modeled according to conspecific flies, whereas condition scaled with facial saturation in females. We found no such relationships for their wing interference pattern nor abdomens, with the latter included as a nonsexual control. Our results suggest that the structurally colored faces, but not the iridescent wings, of male and female L. cana are reliable guides to individual quality and support the broader potential for structural colors as honest signals. They also highlight the potential for mutual mate choice in this system, while arguing for 1 of several alternate signaling roles for wing interferences patterns among the myriad taxa which bear them. [ABSTRACT FROM AUTHOR]

Liver Kinase B1 (LKB1) is known as a master kinase for 14 kinases related to the adenosine monophosphate-activated protein kinase. Two of them salt inducible kinase 3 and adenosine monophosphate-activated protein kinase a have previously been implicated in sleep regulation. We generated loss-of-function mutants for Lkb1 in both Drosophila and mice. Sleep, but not circadian rhythms, was reduced in Lkb1-mutant flies and in flies with neuronal deletion of Lkb1. Genetic interactions between Lkb1 and threonine to alanine mutation at residue 184 of adenosine monophosphate-activated protein kinase in Drosophila sleep or those between Lkb1 and Threonine to Glutamic Acid mutation at residue 196 of salt inducible kinase 3 in Drosophila viability have been observed. Sleep was reduced in mice after virally mediated reduction of Lkb1 in the brain. Electroencephalography analysis showed that nonrapid eye movement sleep and sleep need were both reduced in Lkb1-mutant mice. These results indicate that liver kinase B1 plays a physiological role in sleep regulation conserved from flies to mice. [ABSTRACT FROM AUTHOR]

Due to the increase in global transport in recent decades, species that flourish in human-altered environments are widening their geographical distribution. Consequently, endemic species are either declining or going to extinction. Here, we report the first occurrence of the Afrotropical fly Zaprionus tuberculatus Malloch in the Americas. This species has been recognized as invasive by the Invasive Species Compendium, and we argue that it will probably impact drosophilid communities. Our study was conducted on the natural and urban environments of the Brazilian Savanna, a biodiversity hotspot where drosophilid communities have been monitored since 1999. Z. tuberculatus was first collected in January 2020, at low abundances, in urban parks located in Brasília. In December 2020, we recorded it in a preserved area approximately 200 km away from the urban parks. From January to March 2021, we found the species in seven urban parks in Brasília and three natural reserves (conservation units) located around the city. The species' relative abundance increased from 0.9% in 2020 (n = 11,244 drosophilids) to 17% in 2021 (n = 6,002 drosophilids). This is a rare opportunity to monitor a recent invasion event in a well-studied area. Based on the impact of Z. indianus Gupta, which invaded the Brazilian Savanna in 1999 and remained the dominant species during the rainy seasons, we fear that this new arrival will reduce the diversity of drosophilids in the Neotropics. [ABSTRACT FROM AUTHOR]

Organic yard waste from western Washington, U.S. that may contain puparia of apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae), had been moved to central Washington for composting, threatening the $3 billion apple industry concentrated in that region. Heating waste to kill fly puparia before it is transported could be a solution to this problem. Here, we report results of studies in 2016–2021 that sought to identify a minimum heat treatment simulating that obtained using a low-pressure steam generator for maximizing kill of R. pomonella puparia. In two experiments, puparia were exposed to temperatures ramped linearly over 6 h from 21°C to 47.8, 51.1, 55.0, or 60.0°C in an oven. The 47.8, 51.1, and 55°C treatments did not achieve 100% mortality, although only one adult fly from 4,000 puparia was found in the 55°C treatment, while no puparia survived the 60°C treatment. In a third, similar experiment, no puparia out of 2,400 exposed to 55°C survived. In a fourth and final experiment conducted over 3 years, no puparia out of 61,223 exposed to a 6-h ramp from 21°C to 55°C followed by a 1-h hold time at 55°C produced flies. In addition, all puparia in this treatment died. Based on 42.3 to 69.8% control survival, 31,217 puparia were killed by this treatment with no survivors, for a probit 8.7190 level of security. Results suggest that the 55°C and 1-h hold time treatment here is close to the minimum heat regime needed for disinfesting organic waste of R. pomonella puparia. [ABSTRACT FROM AUTHOR]

Inflammaging refers to low-grade, chronically activated innate immunity that has deleterious effects on healthy lifespan. However, little is known about the intrinsic signaling pathway that elicits innate immune genes during aging. Here, using Drosophila melanogaster, we profile the microRNA targetomes in young and aged animals, and reveal Dawdle, an activin-like ligand of the TGF-β pathway, as a physiological target of microRNA-252. We show that microRNA-252 cooperates with Forkhead box O, a conserved transcriptional factor implicated in aging, to repress Dawdle. Unopposed Dawdle triggers hyperactivation of innate immune genes coupled with a decline in organismal survival. Using adult muscle tissues, single-cell sequencing analysis describes that Dawdle and its downstream innate immune genes are expressed in distinct cell types, suggesting a cell nonautonomous mode of regulation. We further determine the genetic cascade by which Dawdle signaling leads to increased Kenny/IKKc protein, which in turn activates Relish/NF-jB protein and consequentially innate immune genes. Finally, transgenic increase of microRNA-252 and Forkhead box O pathway factors in wild-type Drosophila extends lifespan and mitigates the induction of innate immune genes in aging. Together, we propose that microRNA-252 and Forkhead box O promote healthy longevity by cooperative inhibition on Dawdle-mediated inflammaging. [ABSTRACT FROM AUTHOR]

Cluster flies are represented by the genus Pollenia Robineau-Desvoidy, 1830 of the family Polleniidae Brauer and Bergenstamm, 1889. Their larvae are known to be internal parasites or predators of earthworms. Herein, we report for the first time the occurrence of the cluster flies Pollenia rudis Fabricius, 1794 and Pollenia vagabunda (Meigen, 1826) (Diptera: Polleniidae) on carcasses in Algeria and identify them through DNA barcoding. A region of the mitochondrial cytochrome c oxidase I gene (COI) was amplified and sequenced. Genetic distances were determined. A phylogenetic tree was constructed with the maximum parsimony method using 10 000 bootstrap replicates. A total number of 157 adults of P. rudis were collected together with 325 adults of Pollenia vagabunda. The occurrence of Pollenia on animal carcasses does not seem to be correlated with a particular stage of decomposition. All the sequences were correctly identified using the BLASTn tool from the GenBank database and the BOLD identification engine. Intra- and interspecific sequence divergence values were less than 1% and greater than 3%, respectively. COI barcodes obtained from this study were robust enough to identify and distinguish unambiguously between P. rudis and P. vagabunda. In the tree-based analysis, the cluster flies were all assigned to their respective species separately from each other confirming the morphological identification. These results provide DNA barcodes that contribute to the growth of reference databases and allow fast and accurate identification. [ABSTRACT FROM AUTHOR]

The invasive avian vampire fly (Philornis downsi, Diptera: Muscidae) is considered one of the greatest threats to the endemic avifauna of the Gala´pagos Islands. The fly larvae parasitize nearly every passerine species, including Darwin’s finches. Most P. downsi research to date has focused on the effects of the fly on avian host fitness and mitigation methods. A lag in research related to the genetics of this invasion demonstrates, in part, the need to develop full-scale genomic resources with which to address further questions within this system. In this study, an adult female P. downsi was sequenced to generate a high-quality genome assembly. We examined various features of the genome (e.g., coding regions and noncoding transposable elements) and carried out comparative genomics analysis against other dipteran genomes. We identified lists of gene families that are significantly expanding or contracting in P. downsi that are related to insecticide resistance, detoxification, and counter defense against host immune responses. The P. downsi genome assembly provides an important resource for studying the molecular basis of successful invasion in the Gala´pagos and the dynamics of its population across multiple islands. The findings of significantly changing gene families associated with insecticide resistance and immune responses highlight the need for further investigations into the role of different gene families in aiding the fly’s successful invasion. Furthermore, this genomic resource provides a necessary tool to better inform future research studies and mitigation strategies aimed at minimizing the fly’s impact on Gala´pagos birds. [ABSTRACT FROM AUTHOR]

Prevention and control programs for Ceratitis capitata require a large supply of lures and traps for use in established trapping networks and mass-trapping suppression measures. The main lures currently used are: Trimedure (TML), three-component Biolure (BL), and Ceratrap (CT). The aim of this study was to determine the release rates of these lures, the chemical composition of their volatiles, and how these parameters change with exposure time. Tests were conducted under field conditions at three different elevations (25, 500, and 1,300 masl) during the dry and rainy seasons in Chiapas, Mexico. We found that for TML and BL, the release rate was similar in both seasons and at all three elevations. In the case of CT, the release rate was greater during the dry season and at the lowest elevation during the rainy season. With the caveat of using solid-phase microextraction technique for identification of lure compounds in this study, we found that the volatile compounds of TML were maintained throughout the rainy season, however, in the dry season, some compounds could not be detected. The volatile compounds emitted by BL were trimethylamine, ammonium acetate, and acetamide. Among volatile compounds of CT, acetic acid was the most abundant in the rainy season, while minor compounds were only detected during the first five weeks. Recapture rates were affected by elevation in the three lures tested and there was a significant interaction between elevation in exposure time for TML and BL. [ABSTRACT FROM AUTHOR]

The black soldier fly (BSF) Hermetia illucens (L.) (Diptera: Stratiomyidae) has been recognized as a promising insect species for sustainable management of organic waste and by-products. Indoor breeding of BSF with artificial lighting has been proved successful, but efforts are still needed to optimize BSF reproductive output. Increasing adult density seems an option to exploit space, whereas decreasing artificial lighting duration may reduce unnecessary power consumption. This study aimed at investigating the effects of adult density (10, 25, and 50 pairs per 30 × 30 × 30 cm cage; i.e., 370, 926, and 1,852 pairs/m3), light regime (8:16, 12:12, and 16:8 [L:D] h), and their possible interactions, on some BSF life history traits relevant to reproduction. The results show that the overall BSF reproductive output increased with increasing adult density but was not affected by light regimes per se. With the highest BSF adult density tested, an average of more than 20,000 neonate larvae were produced from a cage within 10 d. At this density, increasing photoperiod increased neonate production, but also decreased the number of neonates per watt used for artificial illumination. The temporal oviposition patterns, mean individual female reproductive output, mating success, egg hatching rate, and insect survival rate were not affected by adult density or light regime as simple effects. However, the interaction between adult density and light regime was significant for the first oviposition peak, mean individual female reproductive output, and insect survival rate. The possible mechanisms behind our results are discussed. [ABSTRACT FROM AUTHOR]

Wings are probably the most advanced evolutionary novelty in insects. In the fruit fly Drosophila melanogaster, proper development of wings requires the activity of so-called wing hearts located in the scutellum of the thorax. Immediately after the imaginai ecdysis, these accessory circulatory organs remove hemolymph and apoptotic epidermal cells from the premature wings through their pumping action. This clearing process is essential for the formation of functional wing blades. Mutant flies that lack intact wing hearts are flightless and display malformed wings. The embryonic wing heart progenitors originate from two adjacent parasegments corresponding to the later second and third thoracic segments. However, adult dipterian flies harbor only one pair of wings and only one pair of associated wing hearts in the second thoracic segment. Here we show that the specification of WHPs depends on the regulatory activity of the Hox gene Ultrabithorax. Furthermore, we analyzed the development of wing hearts in the famous four-winged Ultrabithorax (Ubx) mutant, which was first discovered by Ed Lewis in the 1970s. In these flies, the third thoracic segment is homeotically transformed into a second thoracic segment resulting in a second pair of wings instead of the club-shaped halteres. We show that a second pair of functional wing hearts is formed in the transformed third thoracic segment and that all wing hearts originate from the wild-type population of wing heart progenitor cells. [ABSTRACT FROM AUTHOR]

Argonaute proteins of the PIWI clade complexed with PIWI-interacting RNAs (piRNAs) protect the animal germline genome by silencing transposable elements. One of the leading experimental systems for studying piRNA biology is the Drosophila melanogaster ovary. In addition to classical mutagenesis, transgenic RNA interference (RNAi), which enables tissue-specific silencing of gene expression, plays a central role in piRNA research. Here, we establish a versatile toolkit focused on piRNA biology that combines germline transgenic RNAi, GFP marker lines for key proteins of the piRNA pathway, and reporter transgenes to establish genetic hierarchies. We compare constitutive, pan-germline RNAi with an equally potent transgenic RNAi system that is activated only after germ cell cyst formation. Stage-specific RNAi allows us to investigate the role of genes essential for germline cell survival, for example, nuclear RNA export or the SUMOylation pathway, in piRNA-dependent and independent transposon silencing. Our work forms the basis for an expandable genetic toolkit provided by the Vienna Drosophila Resource Center. [ABSTRACT FROM AUTHOR]

During the maternal-to-zygotic transition (MZT), which encompasses the earliest stages of animal embryogenesis, a subset of maternally supplied gene products is cleared, thus permitting activation of zygotic gene expression. In the Drosophila melanogaster embryo, the RNA-binding protein Smaug (SMG) plays an essential role in progression through the MZT by translationally repressing and destabilizing a large number of maternal mRNAs. The SMG protein itself is rapidly cleared at the end of the MZT by a Skp/Cullin/F-box (SCF) E3-ligase complex. Clearance of SMG requires zygotic transcription and is required for an orderly MZT. Here, we show that an F-box protein, which we name Bard (encoded by CG14317), is required for degradation of SMG. Bard is expressed zygotically and physically interacts with SMG at the end of the MZT, coincident with binding of the maternal SCF proteins, SkpA and Cullin 1, and with degradation of SMG. shRNA-mediated knock-down of Bard or deletion of the bard gene in the early embryo results in stabilization of SMG protein, a phenotype that is rescued by transgenes expressing Bard. Bard thus times the clearance of SMG at the end of the MZT. [ABSTRACT FROM AUTHOR]

Plastic recombination in Drosophila melanogaster has been associated with a variety of extrinsic and intrinsic factors such as temperature, starvation, and parasite infection. The bacterial endosymbiont Wolbachia pipientis has also been associated with plastic recombination in D. melanogaster. Wolbachia infection is pervasive in arthropods and this infection induces a variety of phenotypes in its hosts, the strength of which can depend on bacterial titer. Here, we test the hypothesis that the magnitude of Wolbachia-associated plastic recombination in D. melanogaster depends on titer. To manipulate titer, we raised Wolbachia-infected and uninfected flies on diets that have previously been shown to increase or decrease Wolbachia titer relative to controls. We measured recombination in treated and control individuals using a standard backcrossing scheme with two X-linked visible markers. Our results recapitulate previous findings that Wolbachia infection is associated with increased recombination rate across the yellow-vermillion interval of the X chromosome. Our data show no significant effect of diet or diet by Wolbachia interactions on recombination, suggesting that diet-induced changes in Wolbachia titer have no effect on the magnitude of plastic recombination. These findings represent one of the first steps toward investigating Wolbachia-associated plastic recombination and demonstrate that the phenotype is a discrete response rather than a continuous one. [ABSTRACT FROM AUTHOR]

Tolerance of terrestrial insects in temperate regions to water immersion and hypoxia has rarely been studied but can be an important adaptation to moist environments, with implications for insect dispersal through waterways. In the Pacific Northwest of the United States, apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera:Tephritidae), can be found in riparian habitats subject to flooding. Here, survival of R. pomonella larvae and different age puparia after flotation or immersion in 13.3°C or 21.1°C water for 1–12 d was determined. Larvae sank in water and when submerged for 1 or 2 d suffered greater mortality than control larvae. Fewer young (1–2 d old) than older puparia (13–15 d old) floated in water. When immersed in water for 1–12 d, young puparia suffered greater mortality than older puparia, which were not affected by water immersion. Consequently, fewer adult flies eclosed from puparia that had been water treated when young than older. Adult flies from pre-chill and post-chill puparia that had been water treated eclosed later than control flies, but treatment flies survived about 60 d and reproduced. Although newly-formed puparia are susceptible to hypoxic water conditions, increased buoyancy and water tolerance occur rapidly after formation, perhaps making survival possible and allowing water-borne dispersal of older puparia. [ABSTRACT FROM AUTHOR]

Larval characters are of importance in systematic and evolutionary studies of Diptera but lag behind characters of adults due to difficulties in obtaining relevant information. Larvae of stomach bot flies are obligate parasites completing development exclusively in the alimentary tract of equids and rhinoceroses. They possess diversified morphological adaptations, providing remarkable examples to further our understanding of larval evolution. Herein, three-dimensional structures of first instar Gasterophilus pecorum and Gyrostigma rhinocerontis are compared using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). We suggest CLSM has a large potential for exploiting cryptic character systems of micro fly larvae, as spectral range and intensity of autofluorescence emitted by sclerotized structures and soft tissues are distinct, presenting a high-contrast mechanism for multistructural visualization with non-destructive sample preparation. Five new potential synapomorphies are proposed to corroborate the sister-group Gasterophilus and Gyrostigma. The upward curving mouth-hooks of first instar Gasterophilus and Gyrostigma are distinctive in Cyclorrhapha and possibly serve to facilitate the larval subcutaneous migration within the host. Three types of mouthhooks are recognized in first instar Oestridae, with the gently curved and gradually tapered type optimized as the ancestral state, from which the gasterophiline and hypodermatine types evolved independently. [ABSTRACT FROM AUTHOR]

Black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae), larvae are suspected to inhibit house flies, Musca domestica (L.) (Diptera: Muscidae), seeking oviposition sites. However, the mechanisms regulating these interactions are not well known. In this study, a Y-tube olfactometer was used to determine whether black soldier fly eggs, larvae, puparia (without pupae inside), adult carcasses, or resulting frass of black soldier fly decrease attractiveness of adult house flies to an oviposition substrate (i.e. sugar and wheat bran diet, used as the control). Secondly, two symbiotic bacteria associated were isolated, identified, and tested for house fly attractiveness with a Y-tube olfactometer. Thirdly, volatile organic compounds produced by swine feed before and after black soldier fly, or house fly larval digestion were analyzed with gas chromatography–mass spectrometry (GC–MS). Results indicate feed mixed with black soldier fly larval frass, and symbiotic bacteria, Paenalcaligenes sp. associated with black soldier fly, were less attractive to house flies than the control. However, another symbiotic bacterium Lysinibacillus sp. associated with black soldier fly pupae was attractive. In total, 52 volatile organic compounds were identified from treatments previously described. Quantitative differences in volatile profiles across treatments potentially explain the mechanism regulating house fly substrate choices in the presence, or absence, of black soldier fly larvae. [ABSTRACT FROM AUTHOR]

The sterile insect technique (SIT) is a sustainable pest management tool based on the release of millions of sterile insects that suppress reproduction in targeted populations. Success of SIT depends on survival, maturation, dispersal, and mating of released sterile insects. Laboratory and field cage studies have demonstrated that dietary supplements of methoprene and raspberry ketone (RK) promote sexual maturation of adult Queensland fruit fly, Bactrocera tryoni (Froggatt), and may hence shorten the delay between release and maturity in the field.We investigated the effects of methoprene and RK dietary supplements on field abundance of sexually mature sterile Q-flies relative to untreated flies fed only sugar and yeast hydrolysate before release at 2 d of age. Compared with untreated flies, more methoprene- and RK-treated flies were recaptured in cuelure traps to which only sexually mature males are attracted. At distances of 100 and 200 m from the release point, recapture rates were higher for methoprene- and RK-treated flies than for untreated flies, but at 300 m recapture rates were low and were similar for treated and untreated flies. Rainfall, relative humidity, wind speed, and wind direction did not affect recapture rates, but temperature was positively correlated with recapture rates for all treatments. There was a strong correlation between the number of sterile and wild flies caught in traps, indicating co-location in the field. Dietary supplements of methoprene and RK can substantially increase abundance of sexually mature sterile male Q-flies in the field following release as 2-d-old immature adults. [ABSTRACT FROM AUTHOR]

Some leafmining fly species are pests of agricultural and ornamental plants but they also include many species with little economic importance. The taxonomy of leafmining flies is often challenging because of putative cryptic species. Following new outbreaks of Liriomyza (Diptera:Agromyzidae) in Australia, we here characterize 13 dipteran leafminer species collected from Australia based on cytochrome c oxidase subunit 1 (COI) DNA barcodes, and we compare these with overseas data. We confirm barcodes of nine species from the Agromyzidae (Liriomyza sativae, L. huidobrensis, L. trifolii, L. bryoniae, L. chinensis, L. brassicae, L. chenopodii, Phytomyza plantaginis and P. syngenesiae) and one species from the Drosophilidae (Scaptomyza flava); we describe new haplotypes for some of these species. Furthermore, we provide the first DNA barcodes for two species (Cerodontha milleri and Phytoliriomyza praecellens) from the Agromyzidae and one species (Scaptomyza australis) from the Drosophilidae. These findings provide a baseline for DNA-based identification of pest Liriomyza incursions spreading across the Australian east coast in relation to other species already present in Australia. [ABSTRACT FROM AUTHOR]

Cholinergic signaling dominates the insect central nervous system, contributing to numerous fundamental pathways and behavioral circuits. However, we are only just beginning to uncover the diverse roles different cholinergic receptors may play. Historically, insect nicotinic acetylcholine receptors have received attention due to several subunits being key insecticide targets. More recently, there has been a focus on teasing apart the roles of these receptors, and their constituent subunits, in native signaling pathways. In this study, we use CRISPRCas9 genome editing to generate germline and somatic deletions of the Db1 nicotinic acetylcholine receptor subunit and investigate the consequences of loss of function in Drosophila melanogaster. Severe impacts on movement, male courtship, longevity, and wing expansion were found. Loss of Dβ1 was also associated with a reduction in transcript levels for the wing expansion hormone bursicon. Neuronspecific somatic deletion of Dβ1 in bursicon-producing neurons (CCAP-GAL4) was sufficient to disrupt wing expansion. Furthermore, CCAP-GAL4-specific expression of Dβ1 in a germline deletion background was sufficient to rescue the wing phenotype, pinpointing CCAP neurons as the neuronal subset requiring Db1 for the wing expansion pathway. Db1 is a known target of multiple commercially important insecticides, and the fitness costs exposed here explain why field-isolated target-site resistance has only been reported for amino acid replacements and not loss of function. This work reveals the importance of Db1-containing nicotinic acetylcholine receptors in CCAP neurons for robust bursicon-driven wing expansion. [ABSTRACT FROM AUTHOR]

Diptera is one of the biggest insect orders and displays a large diversity of visual adaptations. Similarly to other animals, the dipteran visual process is mediated by opsin genes. Although the diversity and function of these genes are well studied in key model species, a comprehensive comparative genomic study across the dipteran phylogeny is missing. Here we mined the genomes of 61 dipteran species, reconstructed the evolutionary affinities of 528 opsin genes, and determined the selective pressure acting in different species. We found that opsins underwent several lineage-specific events, including an independent expansion of Long Wave Sensitive opsins in flies and mosquitoes, and numerous family-specific duplications and losses. Both the Drosophila and the Anopheles complement are derived in comparison with the ancestral dipteran state. Molecular evolutionary studies suggest that gene turnover rate, overall mutation rate, and site-specific selective pressure are higher in Anopheles than in Drosophila. Overall, our findings indicate an extremely variable pattern of opsin evolution in dipterans, showcasing how two similarly aged radiations, Anopheles and Drosophila , are characterized by contrasting dynamics in the evolution of this gene family. These results provide a foundation for future studies on the dipteran visual system. [ABSTRACT FROM AUTHOR]

Changes in gene regulation at multiple levels may comprise an important share of the molecular changes underlying adaptive evolution in nature. However, few studies have assayed within- and between-population variation in gene regulatory traits at a transcriptomic scale, and therefore inferences about the characteristics of adaptive regulatory changes have been elusive. Here, we assess quantitative trait differentiation in gene expression levels and alternative splicing (intron usage) between three closely related pairs of natural populations of Drosophila melanogaster from contrasting thermal environments that reflect three separate instances of cold tolerance evolution. The cold-adapted populations were known to show population genetic evidence for parallel evolution at the SNP level, and here we find evidence for parallel expression evolution between them, with stronger parallelism at larval and adult stages than for pupae. We also implement a flexible method to estimate cis- vs trans-encoded contributions to expression or splicing differences at the adult stage. The apparent contributions of cis- vs trans-regulation to adaptive evolution vary substantially among population pairs. While two of three population pairs show a greater enrichment of cis-regulatory differences among adaptation candidates, trans-regulatory differences are more likely to be implicated in parallel expression changes between population pairs. Genes with significant cis-effects are enriched for signals of elevated genetic differentiation between cold- and warm-adapted populations, suggesting that they are potential targets of local adaptation. These findings expand our knowledge of adaptive gene regulatory evolution and our ability to make inferences about this important and widespread process. [ABSTRACT FROM AUTHOR]

In some eukaryotes, germline soma differentiation involves elimination of parts of the genome from somatic cells. The portions of the genome restricted to the germline often contain genes that play a role in development and function of the germline. Lineages with germline-restricted DNA are taxonomically diverse, and the size of the germline-restricted genome varies substantially. Unfortunately, few of these lineages have been studied in detail. As a result, we understand little about the general evolutionary forces that drive the origin and maintenance of germline-restricted DNA. One of the taxonomic groups where germline-restricted DNA has been poorly studied are the flies (Diptera). In three Dipteran families, Chironomidae, Cecidomyiidae, and Sciaridae, entire chromosomes are eliminated from somatic cells early in embryonic development. Germline-restricted chromosomes are thought to have evolved independently in the Dipteran families and their size, number, and transmission patterns vary between families. Although there is a wealth of cytological studies on these chromosomes in flies, almost no genomic studies have been undertaken. As a result, very little is known about how and why they evolved and what genes they encode. This review summarizes the literature on germline-restricted chromosomes in Diptera, discusses hypotheses for their origin and function, and compares germline-restricted DNA in Diptera to other eukaryotes. Finally, we discuss why Dipteran lineages represent a promising system for the study of germline-restricted chromosomes and propose future avenues of research on this topic. [ABSTRACT FROM AUTHOR]

The larva of stone leek leafminer, Liriomyza chinensis (Kato), is known to infest alternately just below the epidermis and inner surface of hollow cylindrical leaves of allium crops, resulting in the formation of discontinuous linear mines (mine form: discontinuous). However, after the fall of 2016, a novel mine form of the same species (mine form: continuous) was detected in Welsh onion fields of Kyoto Prefecture, Japan. We hypothesized that these mine forms were associated with flies having different genetic backgrounds; hence, we compared the mine forms and the partial mtCOI gene of flies collected from Welsh onion fields from 2018 to 2019. The results demonstrated that the flies that emerged from different mine forms could be classified into two haplogroups, i.e. flies displaying a discontinuous mine form were of haplogroup A, whereas those that displayed continuous mines were of haplotype B. Additionally, using populations of these flies reared in the laboratory, we confirmed that the mine form of the larvae of haplotype A on Welsh onions was discontinuous, whereas that of haplotype B was continuous. We named the population that exhibited a discontinuous mine form as biotype A and the population displaying a continuous mine form as biotype B. [ABSTRACT FROM AUTHOR]

The family Tachinidae ("bristle flies") is the most diverse and ecologically important group of insect parasitoids outside the parasitic wasps. It is among the most species rich families of flies (Diptera) and has experienced a recent adaptive radiation across the globe. We make use of a molecular phylogeny of the family to examine its rapid radiation and explore the traits of tachinid lineages that may have contributed to variation in their diversification. We apply a range of diversification analyses to assess the consistency and robustness of effects. We find that the Tachinidae are among the most rapidly diversifying families of animals. Six to eight clades of bristle flies, distributed across the phylogeny, exhibit strong evidence of accelerated diversification. Our results suggest that the use of holometabolous insect larvae, and specifically caterpillars (Lepidoptera), as hosts, is associated with increased diversification rates. However, these effects were inconsistent across analyses. We detected little influence of oviposition strategy (egg type) or host feeding habit, and we recovered evidence that unmeasured "hidden" traits may explain greater variance in diversification. We evaluated the strengths and weaknesses of different Maximum Likelihood and Bayesian approaches for analysing diversification and the potential for extrinsic factors, such as geography, to influence patterns of richness and diversification. In general, we conclude that although certain traits may provide opportunities for diversification, whether this is capitalized on may depend on additional traits and/or historical contingency. [ABSTRACT FROM AUTHOR]

The house fly, Musca domestica L. is an international nuisance and vector of numerous pathogens that cause disease syndromes in humans and animals. In urban areas, fly exclusion is a major part of the pest management program at many commercial establishments. If flies elude the exclusion techniques, a system for fly management inside the building is the next line of defense. Permission was given to evaluate the fly management system consisting of 12 GT-180 and 6 MX-360 ultraviolet (UV) light traps inside of a large supermarket for 8 wk. GT-180 traps mounted 2 m high on walls captured significantly fewer flies than MX-360s, larger portable traps placed on the floor. Both glue boards in GT-180 traps captured similar fly numbers. In the MX-360 traps, mean fly numbers captured by the two vertically mounted glue boards combined were not significantly different from those captured by the single lower horizontal glue board. Changing from white to black glue boards significantly reduced fly numbers captured by both traps. Three of the six traps capturing the largest mean fly numbers were in the Bakery, followed by the Deli. A total of 3,626 house flies (453/week) was captured during the study. Changes in weekly fly populations inside the store were visibly obvious because fly numbers were so high. Difference in flies trapped because of trap location and height, and glue board color are further discussed. [ABSTRACT FROM AUTHOR]

Wing interference patterns (WIPs) are stable structural colours displayed on insect wings which are only visible at specific viewing geometries and against certain backgrounds. These patterns are widespread among flies and wasps, and growing evidence suggests that they may function as species‐ and sex‐specific mating cues in a range of taxa. As such, it is expected that WIPs should differ between species and show clear sexual dimorphisms. However, the true extent to which WIPs vary between species, sexes and individuals is currently unclear, as previous studies have only taken a qualitative approach, without considering how WIPs might be perceived by the insect. Here, we perform the first quantitative analysis of inter‐ and intra‐specific variation in WIPs across seven Australian species of the blowfly genus Chrysomya. Using multispectral digital imaging and a tentative model of blowfly colour vision, we provide quantitative evidence that WIPs are species‐specific, highlight that the extent of divergence is greater in males than in females and demonstrate sexual dimorphisms in several species. These data suggest that WIPs have diversified substantially in blowflies as a result of either sexual or ecological selection. [ABSTRACT FROM AUTHOR]