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Aeshnidae Rambur, 1842 are impressive large insects distributed worldwide. Currently, over 500 species are recognized. Nevertheless, the phylogeny of this family is not completely understood. We applied molecular phylogenetic analysis using two popular phylogenetic markers, the mitochondrial COI gene fragment (barcoding sequence) and the nucleic ITS region, containing the ITS1, 5.8S rRNA, and ITS2 sequences. We used available and credible published sequences and 96 newly sequenced specimens. Our analysis involved all West Palaearctic species, all but one genera of the Holarctic Aeshnidae, and most genera worldwide, and is by far the largest molecular study of this family. The topology of all trees created with different algorithms and genes is in favour of the current taxonomic concept, with some remarkable outcomes. Aeshna Fabricius, 1775, was found to be diverged into several branches, especially with respect to the COI gene. Although it appeared not monophyletic in phylogenetic reconstructions based on the ITS region, the analysis of COI and joint analysis suggest its monophyly in the current taxonomical sense, with one notable exception. Aeshna isoceles (Müller, 1767) has fallen out of Aeshna in all analyses, so a new monophyletic genus, Isoaeschna gen. nov. is introduced for it. The genus Brachytron Evans, 1845 tightly clustered with Aeschnophlebia Selys, 1883, Epiaeschna Hagen in Selys, 1883, and Nasiaeschna Selys in Förster, 1900. Thus, we suggest subsuming these four genera under the priority name Brachytron. Tetracanthagyna Selys, 1883 clusters as expected with Brachytron in the ITS tree, but is an independent ancient clade of its own in all COI trees. The genus Polycanthagyna Fraser, 1933 syn. nov. is synonymised to Indaeschna Fraser, 1926. On the species level, we suggest that the American Aeshna septentrionalis Burmeister, 1839 be treated as a subspecies of A. caerulea (Ström, 1783), Aeshna caerulea septentrionalis. We synonymize Gynacantha hyalina Selys, 1882 with Gynacantha subinterrupta Rambur, 1842. Our analysis provides new insights on the tight relationships of the circumboreal species Aeshna juncea and A. subarctica and the intraspecies phylogeny of Aeshna juncea.

Aeshna mixta (Latreille 1805) is added to the Anisoptera fauna of Pakistan by reporting it from district Neelum of Azad Jammu and Kashmir, Pakistan. Geopolitically the study area lies in an area facing uncertain ground conditions since inception of Pakistan. It is a hard to reach valley between India and Pakistan, very close to the line of control. Out of twenty sampling sites, specimens of Aeshna mixta were found from fifteen localities of the district. With the addition of this taxon, Anisoptera fauna of Pakistan now count 74 species. The area under district Neelum represents many lush green valleys with lots of natural water bodies that support a broad complex of Odonata. More surveys in the area are suggested to unveil more important records from the area.

Research on the Awash River focused on the upper section, while the middle and lower regions received little attention. Thus, the goal of this study was to evaluate the impact of anthropogenic activities on the upper and middle Awash River. The study took place in nine sampling locations in dry and wet seasons from September 2021 to April 2022 using a multi-habitat sampling approach. We used macroinvertebrate metrics, environmental variables, and multivariate analysis to evaluate ecological health. The highest concentrations of NO 3 , soluble reactive phosphorus, and total phosphorus (0.50–0.93 mg L −1 ) were recorded at the river-mouth of the upper Awash, while the locations below Metehara had the lowest levels of dissolved oxygen (1.81–2.33 mg L −1 ). Redundancy analysis indicated that dissolved oxygen, NH 3 , temperature, NO 2 , pH, TSS, NO 3 , and TDS influenced macroinvertebrate distribution. The presence of the sensitive groups Caenidae, Hydropsychidae, Heptageniidae, and Aeshnidae at upstream sites indicated better ecological conditions. The middle and downstream sites supported moderately tolerant and tolerant taxa demonstrating water quality impairment. The lowest Ethiopian biotic score was recorded at the river-mouth of the upper Awash. The study sites below Metehara demonstrated severe ecological impairment since highly tolerant taxa were abundant and had strong correlations with temperature, TSS, and TDS levels. Pollutants from agricultural farms and domestic and industrial wastes from Addis Ababa, Metehara, and Merti towns most likely affect the impaired sites. This study demonstrated that the middle Awash experienced substantial ecological deterioration, indicating the need for restoration works to fit the water for socio-economic development.

(1) To increase awareness of the challenges induced by imperfect detection, which is a fundamental issue in species distribution modelling; (2) to emphasize the value of replicate observations for species distribution modelling; and (3) to show how 'cheap' checklist data in faunal/floral databases may be used for the rigorous modelling of distributions by site-occupancy models. Switzerland. We used checklist data collected by volunteers during 1999 and 2000 to analyse the distribution of the blue hawker, Aeshna cyanea (Odonata, Aeshnidae), a common dragonfly in Switzerland. We used data from repeated visits to 1-ha pixels to derive 'detection histories' and apply site-occupancy models to estimate the 'true' species distribution, i.e. corrected for imperfect detection. We modelled blue hawker distribution as a function of elevation and year and its detection probability of elevation, year and season. The best model contained cubic polynomial elevation effects for distribution and quadratic effects of elevation and season for detectability. We compared the site-occupancy model with a conventional distribution model based on a generalized linear model, which assumes perfect detectability (p = 1). The conventional distribution map looked very different from the distribution map obtained using site-occupancy models that accounted for the imperfect detection. The conventional model underestimated the species distribution by 60%, and the slope parameters of the occurrence-elevation relationship were also underestimated when assuming p = 1. Elevation was not only an important predictor of blue hawker occurrence, but also of the detection probability, with a bell-shaped relationship. Furthermore, detectability increased over the season. The average detection probability was estimated at only 0.19 per survey. Conventional species distribution models do not model species distributions per se but rather the apparent distribution, i.e. an unknown proportion of species distributions. That unknown proportion is equivalent to detectability. Imperfect detection in conventional species distribution models yields underestimates of the extent of distributions and covariate effects that are biased towards zero. In addition, patterns in detectability will erroneously be ascribed to species distributions. In contrast, site-occupancy models applied to replicated detection/non-detection data offer a powerful framework for making inferences about species distributions corrected for imperfect detection. The use of 'cheap' checklist data greatly enhances the scope of applications of this useful class of models.

Abstract Odonates are important biological control agents for the control of insect pests and insect disease vectors of medical and veterinary importance. The present study was conducted to evaluate the odonate fauna of Swat, Pakistan from March to October 2019. A total of 200 specimens of odonates were collected from diverse habitats. The collected specimens of the order Odonata belonged to 5 families, three families of suborder Anisoptera namely Libellulidae, Gomphidae and Aeshnidae while two families of suborder Zygoptera (Chlorocyphidae and Coenagrionidae). The specimens were categorized into 12 genera and 22 species. Libellulidae was the dominant family (n = 138) accounting for 69% of the odonate fauna. Orthetrum was the dominant genus (n = 73) of suborder Anisoptera accounting for 36.5% of the odonate fauna. The least dominant genera were Anax, Paragomphus and Rhyothemis (n = 5 each) accounting each for 2.5% of the odonate fauna. In Zygoptera, the dominant genus was Ceriagrion (12.5%) and the least dominant genus was Ischnura (6%). Pantala flavescens (Fabricius, 1798) was the most abundant odonate species in the study area recorded from all surveyed habitats. Shannon Diversity Index (H) was 2.988 and Simpson Diversity Index (D) was 0.95 for the collected odonate fauna. The highest abundance of Odonata was recorded in August, September and May while no odonate species were recorded in January, February, November and December. Lotic water bodies were the most suitable habitats with abundant odonate fauna. Anax immaculifrons (Rambur, 1842) was the largest sized odonate species having a wingspan of 53.2±1.63 mm and body length of 56.3 ± 0.4 mm. The present study shows the status of odonate fauna of Swat, Pakistan in diverse habitats and seasonsonal variation throughout the year. Further work is recommended to bridge the gaps in the existing literature. Resumo Odonatos são importantes agentes de controle biológico para o controle de insetos-praga e vetores de doenças de insetos de importância médica e veterinária. O presente estudo foi conduzido para avaliar a fauna de odonatos de Swat, Paquistão, de março a outubro de 2019. Um total de 200 espécimes de odonatos foi coletado em diversos habitats. Os espécimes coletados da ordem Odonata pertenciam a cinco famílias, três famílias da subordem Anisoptera, a saber, Libellulidae, Gomphidae e Aeshnidae, enquanto duas famílias eram da subordem Zygoptera (Chlorocyphidae e Coenagrionidae). Os espécimes foram classificados em 12 gêneros e 22 espécies. Libellulidae foi a família dominante (n = 138), respondendo por 69% da fauna de odonatos. Orthetrum foi o gênero dominante (n = 73) da subordem Anisoptera, responsável por 36,5% da fauna de odonatos. Os gêneros menos dominantes foram Anax, Paragomphus e Rhyothemis (n = 5 cada), representando cada um 2,5% da fauna de odonatos. Em Zygoptera, o gênero dominante foi Ceriagrion (12,5%), e o gênero menos dominante foi Ischnura (6%). Pantala flavescens (Fabricius, 1798) foi a espécie de odonato mais abundante na área de estudo, registrada em todos os habitats pesquisados. O Índice de Diversidade de Shannon (H) foi de 2,988, e o Índice de Diversidade de Simpson (D) foi de 0,95 para a fauna de odonatos coletados. A maior abundância de Odonata foi registrada em agosto, setembro e maio, enquanto nenhuma espécie de Odonata foi registrada em janeiro, fevereiro, novembro e dezembro. Corpos d’água lóticos foram os habitats mais adequados, com abundante fauna de odonatos. Anax imaculifrons (Rambur, 1842) foi a espécie de odonato de maior tamanho, com envergadura de 53,2 ± 1,63 mm e comprimento do corpo de 56,3 ± 0,4 mm. O presente estudo mostrou o status da fauna de odonatos de Swat, Paquistão, em diversos habitats e variação sazonal ao longo do ano. Recomenda-se trabalho adicional para preencher as lacunas na literatura existente.

Indirect species interactions are ubiquitous in nature, often outnumbering direct species interactions. Yet despite evidence that indirect interactions have strong ecological effects, relatively little is known about whether they can shape adaptive evolution by altering the strength and/or direction of natural selection. We tested whether indirect interactions affect the strength and direction of pollinator-mediated selection on floral traits of the bumble-bee pollinated wildflower Lobelia siphilitica. We estimated the indirect effects of two pollinator predators with contrasting hunting modes: dragonflies (Aeshnidae and Corduliidae) and ambush bugs (Phymata americana, Reduviidae). Because dragonflies are active pursuit predators, we hypothesized that they would strengthen pollinator-mediated selection by weakening plant–pollinator interactions (i.e., a density-mediated indirect effect). In contrast, because ambush bugs are sit-and-wait predators, we hypothesized that they would weaken or reverse the direction of pollinator-mediated selection by altering pollinator foraging behavior (i.e., a trait-mediated indirect effect). Specifically, if ambush bugs hunt from plants with traits that attract pollinators (i.e., prey), then pollinators will spend less time visiting those plants, weakening or reversing the direction of selection on attractive floral traits. We did not find evidence that high dragonfly abundance strengthened selection on floral traits via a density-mediated indirect effect: neither pollen limitation (a proxy for the strength of plant–pollinator interactions) nor directional selection on floral traits of L. siphilitica differed significantly between high- and low-dragonfly abundance treatments. In contrast, we did find evidence that ambush bug presence affected selection on floral traits via a trait-mediated indirect effect: ambush bugs hunted from L. siphilitica plants with larger daily floral displays, reversing the direction of pollinator-mediated selection on daily display size. These results suggest that indirect species interactions have the potential to shape adaptive evolution by altering natural selection.

Macroinvertebrate predators such as backswimmers (Heteroptera: Notonectidae), dragonflies (Odonata: Aeshnidae), and predatory diving beetles (Coleoptera: Dytiscidae) naturally inhabit aquatic ecosystems. Some aquatic ecosystems inhabited by these macroinvertebrate predator taxa equally form malaria vector larval habitats. The presence of these predators in malaria vector larval habitats can negatively impact on development, adult body size, fecundity, and longevity of the malaria vectors, which form important determinants of their fitness and future vectorial capacity. These potential negative impacts caused by aquatic macroinvertebrate predators on malaria vectors warrant their consideration as biocontrol agents in an integrated program to combat malaria. However, the use of these macroinvertebrate predators in malaria biocontrol is currently constrained by technical bottlenecks linked to their generalist predatory tendencies and often long life cycles, demanding complex rearing systems. We reviewed the literature on the use of aquatic macroinvertebrate predators for biocontrol of malaria vectors from the An. gambiae s.l. complex. The available information from laboratory and semi-field studies has shown that aquatic macroinvertebrates have the potential to consume large numbers of mosquito larvae and could thus offer an additional approaches in integrated malaria vector management strategies. The growing number of semi-field structures available in East and West Africa provides an opportunity to conduct ecological experimental studies to reconsider the potential of using aquatic macroinvertebrate predators as a biocontrol tool. To achieve a more sustainable approach to controlling malaria vector populations, additional, non-chemical interventions could provide a more sustainable approach, in comparison with the failing chemical control tools, and should be urgently considered for integration with the current mosquito vector control campaigns. Graphical Abstract

Cytological review of 59 aeshnid species and cytogenetic investigations on Anax ephippiger, Anax immaculifrons, Anax indicus, Anax nigrofasciatus nigrolineatus, Anax parthenope, Gynacantha subinterrupta of the family Aeshnidae by carbol fuchsin staining and C - banding have been under taken. All the species posses 2n = 27m with X0 - XX sex determination except Anax ephippiger with 2n = 14 + neo XY, resulted by the 13 simultaneous fusions among the autosomes and between autosome and sex chromosome. The structure and behaviour of chromosomes, variation in size of m chromosomes and X chromosome and distribution of C - heterochromatin have been studied and compared among the species. C - bands are mostly present at the terminal regions of autosomal bivalents, while Anax ephippiger and Anax parthenope also possess C - bands at the interstitial and sub-terminal regions of the bivalents. Moreover, sex chromosome and m bivalent show variation in distribution of C-heterochromatin in the species. Out of these, chromosome complement of Anax indicus Lieftinck, 1942 and C - banding on Anax ephippiger and Anax indicus have been investigated for the first time. List of cytologically studied species of family Aeshnidae has been updated to 60 species.

Chironomid (Diptera: Chironomidae) larvae play a key role in aquatic food webs as prey for predators like amphibian and dragonfly larvae. This trophic link may be disrupted by anthropogenic stressors such as Bacillus thuringiensis var . israelensis (Bti), a biocide widely used in mosquito control. In a companion study, we recorded a 41% reduction of non-target larval chironomids abundance in outdoor floodplain pond mesocosms (FPMs) treated with Bti. Therefore, we examined the diet of two top predators in the FPMs, larvae of the palmate newt (Salamandridae: Lissotriton helveticus ) and dragonfly (Aeshnidae: predominantly Anax imperator ), using bulk stable isotope analyses of carbon and nitrogen. Additionally, we determined neutral lipid fatty acids in newt larvae to assess diet-related effects on their physiological condition. We did not find any effects of Bti on the diet proportions of newt larvae and no significant effects on the fatty acid content. We observed a trend in Aeshnidae larvae from Bti-FPMs consuming a higher proportion of large prey (Aeshnidae, newt, damselfly larvae; ~42%), and similar parts of smaller prey (chironomid, mayfly, Libellulidae, and zooplankton), compared to controls. Our findings may suggest bottom-up effects of Bti on aquatic predators but should be further evaluated, for instance, by using compound-specific stable isotope analyses of fatty acids or metabarcoding approaches.

Freshwater insects are highly significant as ecosystem service providers, contributing to provisioning services, supporting services, and cultural services. Odonates are dominant predators in many freshwater systems, becoming top predators in fishless ecosystems. One service that odonates provide is the export of matter and energy from aquatic to terrestrial ecosystems. In this study, we provide a review of the literature aiming to estimate the density, biomass, and secondary production of odonates and discuss to what extent this order of insects is relevant for the fertilization of terrestrial ecosystems. We found published data on 109 species belonging to 17 families of odonates from 44 papers. Odonata larvae are abundant in freshwater systems, with a mean density of 240.04 ± 48.01 individuals m−2 (±SE). Lentic habitats show much higher densities (104.40 ± 55.31 individuals m−2, N = 118) than lotic systems (27.12 ± 5.09, N = 70). The biomass estimations for odonates indicate values of 488.56 ± 134.51 mg m−2 y−1, with similar values in lentic and lotic habitats, which correspond to annual secondary productions of 3558.02 ± 2146.80 mg m−2 y−1. The highest biomass is found in dragonflies of the Aeshnidae, Corduliidae, and Gomphidae families. The available evidence suggests a significant potential contribution of Odonata to the exportation of material from water bodies to land. This is further strengthened by the ability of adult odonates to migrate and to colonize different types of water bodies.