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Reference gene AK was identified as a secondary reference gene. In contrast, EF1α was generally ranked in the last or penultimate place. Identification of two stable reference genes will aid in normalizing the expression of target genes for subsequent gene expression studies of X. germanus' mycangium.Natural hybridization is rather widespread and common in animals and can have important evolutionary consequences. In terms of taxonomy, exploring hybridization and introgression is crucial in defining species boundaries and testing taxonomic hypotheses. In the present paper, we report on natural hybrid specimens between Ahlbergia frivaldszkyi (Lederer, 1853) and Callophrys rubi (Linnaeus, 1758). To test the hypothesis of their hybrid origin, we employed the molecular mitochondrial (COI gene) and nuclear (wingless, RPS5, and Ca-ATPase genes) markers commonly used in phylogenetic studies and explored the morphology of the specimens. Our analysis revealed that hybrids bear mitochondrial haplotypes of C. rubi, while nuclear fragments are heterozygous, sharing a combination of A. frivaldszkyi and C. rubi lineages. The hybrid specimens combine morphological characters of both genera. Our results for the first time empirically demonstrate the possibility of genetic introgression between these species and between the genera Callophrys and Ahlbergia on the whole.Life history traits determine the persistence and reproduction of each species. Factors that can affect life history traits are numerous and can be of different origin. We investigated the influence of population origin and heavy metal exposure on microbiota diversity and two life history traits, egg-to-adult viability and developmental time, in Drosophila melanogaster and Drosophila subobscura, grown in the laboratory on a lead (II) acetate-saturated substrate. We used 24 samples, 8 larval and 16 adult samples (two species × two substrates × two populations × two sexes). The composition of microbiota was determined by sequencing (NGS) of the V3-V4 variable regions of the 16S rRNA gene. The population origin showed a significant influence on life history traits, though each trait in the two species was affected differentially. Reduced viability in D. melanogaster could be a cost of fast development, decrease in Lactobacillus abundance and the presence of Wolbachia. The heavy metal exposure in D. subobscura caused shifts in developmental time but maintained the egg-to-adult viability at a similar level. Microbiota diversity indicated that the Komagataeibacter could be a valuable member of D. subobscura microbiota in overcoming the environmental stress. Research on the impact of microbiota on the adaptive response to heavy metals and consequently the potential tradeoffs among different life history traits is of great importance in evolutionary research.The Eocene Baltic amber fossil flies of the genus Acartophthalmites Hennig, 1965 (Diptera Acalyptratae) are revised. Seven species are recognized and described or redescribed. Five species, A. crassipes sp. nov., A. luridus sp. nov., A. rugosus sp. nov., A. tertiaria Hennig, 1965 (type species) and A. willii Pérez-de la Fuente, Hoffeins et Roháček, 2018 are retained in Acartophthalmites while Clusiomites gen. nov. is described for two other species, C. clusioides (Roháček, 2016) comb. nov. (type species) and C. ornatus sp. nov. Relationships of these fossil taxa are discussed and, because they cannot be confidently placed in any known family of Diptera, a new family, Clusiomitidae, is established for them. Clusiomitidae is recognized as a family of Opomyzoidea, probably most closely allied to Clusiidae. These results again confirmed that the diversity of acalyptrate flies was very high in the Mid-late Eocene amber forest.This article introduces the results of a study of three families of Nepomorpha and is the last part of a series of studies that sums up our work on the morphologies of the antennal sensory structures in this taxon. The morphologies and distribution of the sensilla in the families Notonectidae, Pleidae and Helotrephidae were studied under a scanning electron microscope. Six main types (sensilla trichodea, chaetica, campaniformia, basiconica, ampullacea and coeloconica) and ten subtypes (five subtypes of sensilla trichodea and five subtypes of sensilla basiconica) were described. The results were compared with other studies on the antennal sensilla of Nepomorpha in order to assess evolutionary changes within the infraorder. With the use of cladistics analysis, the monophyly of the families Nepidae, Micronectidae, Corixidae and Gelastocoridae was supported. On the other hand, the occurrence of some clades forming superfamilies was weakly supported by bootstrap analysis. These results, supported by presence of the numerous autapomorphies, suggest that antennal sensilla evolved within inner groups.The house fly, Musca domestica L. (Diptera Muscidae), is one of the major vectors of several pathogens that affect humans and animals. We evaluated the toxicity of eight insecticides commonly used for house fly control using five field populations collected from dairies in Riyadh, Saudi Arabia. Among the five tested pyrethroids, non to moderate resistance was found in adults of both sexes compared to a susceptible strain. Resistance ratios ranged from 0.5- to 7-fold for alpha-cypermethrin, 2- to 21-fold for deltamethrin, 4- to 19-fold for bifenthrin, 1- to 9-fold for cyfluthrin, and 1- to 8-fold for cypermethrin. Among the three tested organophosphates, low to moderate resistance was found among adult flies compared to the susceptible strain, and the resistance ratios ranged from 4- to 27-fold for fenitrothion, 2- to 14-fold for chlorpyrifos, and 3- to 12-fold for malathion. The median lethal times for the tested insecticides were 3-33 h for alpha-cypermethrin, 3-24 h for deltamethrin, 5-59 h for bifenthrin, 1-7 h for cypermethrin, 0.3-7 h for cyfluthrin, 6-36 h for fenitrothion, 2-21 h for chlorpyrifos, and 3-34 h for malathion. This study presents baseline data pertaining to registered public health insecticides, and the results will assist future studies monitoring insecticide resistance, and the planning of effective integrated vector management programs.Drought-induced stress and attacks by bark beetle Ips sexdentatus currently result in a massive dieback of Pinus sylvestris in eastern Ukraine. Limited and fragmented knowledge is available on fungi vectored by the beetle and their roles in tree dieback. The aim was to investigate the fungal community vectored by I. sexdentatus and to test the pathogenicity of potentially aggressive species to P. sylvestris. Analysis of the fungal community was accomplished by combining different methods using insect, plant, and fungal material. The material consisted of 576 beetles and 96 infested wood samples collected from six sample plots within a 300 km radius in eastern Ukraine and subjected to fungal isolations and (beetles only) direct sequencing of ITS rDNA. Selleck S3I-201 Pathogenicity tests were undertaken by artificially inoculating three-to-four-year-old pine saplings with fungi. For the vector test, pine logs were exposed to pre-inoculated beetles. In all, 56 fungal taxa were detected, 8 exclusively by isolation, and 13 exclusively by direct sequencing. Those included nine ophiostomatoids, five of which are newly reported as I. sexdentatus associates. Two ophiostomatoid fungi, which exhibited the highest pathogenicity, causing 100% dieback and mortality, represented genera Graphium and Leptographium. Exposure of logs to beetles resulted in ophiostomatoid infections. In conclusion, the study revealed numerous I. sexdentatus-vectored fungi, several of which include aggressive tree pathogens.Terrestrial snails that live in hot and dry climates have developed strategies to cope with high summer temperatures. Several species estivate during the warmest months of the years by resting on vertical supports, typically in groups. Understanding how snails choose their estivation sites and aggregate may lead to the development of new management tools in areas where these snails are invasive. Here, we investigated the preferences of four snail species for vertical supports varying in widths and heights under laboratory and field conditions, and tested whether the presence of conspecifics or snails of other species affected these preferences. The results show that the snails strongly preferred wider supports in laboratory dual-choice tests, and one species (Thebapisana) showed a consistent preference for taller supports as well. These results were confirmed in the field, where more snails were found on wider and taller supports 24 h after being placed in test quadrats. The percentage of snails found in groups on a support was strongly density-dependent. The presence of conspecifics or their mucus did not affect the choices of the snails, nor did the presence of snails of other species or their mucus. Taken together, these results could lead to the development of attractive supports that could be used to mass-capture snails in the field.(1) The project "Tatort Streetlight" implements an insect-friendly road light design in a four year before-after, control-impact (BACI) approach involving citizen scientists. It will broaden the stakeholder interests from solely anthropogenic perspectives to include the welfare of insects and ecosystems. Motivated by the detrimental impacts of road lighting systems on insects, the project aims to find solutions to reduce the insect attraction and habitat fragmentation resulting from roadway illumination. (2) The citizen science approach invites stakeholders to take part and join forces for the development of a sustainable and environmentally friendly road lighting solution. Here, we describe the project strategy, stakeholder participation and motivation, and how the effects of the alternative road luminaire and lighting design can be evaluated. (3) The study compares the changes in (a) insect behavior, (b) night sky brightness, and (c) stakeholder participation and awareness. For this purpose, different experimental areas and stakeholders in four communities in Germany are identified. (4) The project transfers knowledge of adverse effects of improperly managed road illumination and interacts with various stakeholders to develop a new road lighting system that will consider the well-being of street users, local residents, and insects.Anastrepha obliqua (Macquart) (Diptera Tephritidae) is an important pest in the neotropical region. It is considered a polyphagous insect, meaning it infests plants of different taxonomic families and readily colonizes new host plants. The change to new hosts can lead to diversification and the formation of host races. Previous studies investigating the effect of host plants on population structure and selection in Anastrepha obliqua have focused on the use of data from the mitochondrial DNA sequence and microsatellite markers of nuclear DNA, and there are no analyses at the genomic level. To better understand this issue, we used a pooled restriction site-associated DNA sequencing (pooled RAD-seq) approach to assess genomic differentiation and population structure across sympatric populations of Anastrepha obliqua that infest three host plants-Spondias purpurea (red mombin), Mangifera indica (mango) of the family Anacardiaceae and Averrhoa carambola (carambola) of the family Oxalidaceae-in sympatric populations of the species Anastrepha obliqua of Inter-Andean Valley of the Cauca River in southwestern Colombia.