Mandible elemental composition and mechanical properties from distinct castes of the leafcutter ant Atta laevigata (Attini; Formicidae).

Leafcutter ant colonies are divided into castes with the individuals performing different tasks, based mostly on size. With the mandibles, the small minims care for the brood or the fungus, whereas the larger minors and mediae cut and transport plant material, with the ant size positively related to the material size. The mechanical properties and composition of the mandible cuticle have been previously tested in the soldiers as the largest caste, revealing that the cutting edges contained high contents of the cross-linking transition metal zinc (Zn). With regard to the smaller castes, no data are present. To study how the mandible size and function relates to its mechanical properties, we here tested the mandibles of minims, minors and mediae by nanoindentation. We found that the hardness (H) and Young's modulus (E) values increased with increasing ant size and that the mandible cutting edges in each caste have the highest H- and E-values. To gain insight into the origins of these properties, we characterized the elemental composition by energy-dispersive X-ray analysis, revealing that minors and mediae possessed higher content of Zn in the cutting edges in contrast to the minims containing significantly less Zn. This shows, that Zn content relates to higher mechanical property values. Additionally, it shows that all of these parameters can differ within a single species.

Diverse material properties and morphology of moth proboscises relates to the feeding habits of some macromoth and other lepidopteran lineages.

Insects have evolved unique structures that host a diversity of material and mechanical properties, and the mouthparts (proboscis) of butterflies and moths (Lepidoptera) are no exception. Here, we examined proboscis morphology and material properties from several previously unstudied moth lineages to determine if they relate to flower visiting and non-flower visiting feeding habits. Scanning electron microscopy and three-dimensional imaging were used to study proboscis morphology and assess surface roughness patterns on the galeal surface, respectively. Confocal laser scanning microscopy was used to study patterns of cuticular autofluorescence, which was quantified with colour analysis software. We found that moth proboscises display similar autofluorescent signals and morphological patterns in relation to feeding habits to those previously described for flower and non-flower visiting butterflies. The distal region of proboscises of non-flower visitors is brush-like for augmented capillarity and exhibited blue autofluorescence, indicating the possible presence of resilin and increased flexibility. Flower visitors have smoother proboscises and show red autofluorescence, an indicator of high sclerotization, which is adaptive for floral tube entry. We propose the lepidopteran proboscis as a model structure for understanding how insects have evolved a suite of morphological and material adaptations to overcome the challenges of acquiring fluids from diverse sources.

The Microscopic Morphology of Mouthparts and Their Sensilla in the Mycophagous Ladybeetle Illeis chinensis (Coleoptera: Coccinellidae).

The morphological diversity of insect mouthparts is closely related to changes in food sources and diets. Research into the structures of insect mouthparts may help to establish a fundamental basis for a better understanding of insect feeding mechanisms. In this study, we examined the fine morphology of the mouthparts of Illeis chinensis using scanning electron microscopy. We paid particular attention to the types, quantities, and distribution of sensilla on the mouthparts. Our results showed that the basic components of the mouthparts of I. chinensis are the same as those in other lady beetles, i.e., the labrum, mandible, maxillae, labium, and hypopharynx. We also found structural specialization indicating adaptation to fungal feeding. On the mouthparts, there are eight kinds of sensilla and two kinds of glandular structures, including sensilla chaetica, sensilla basiconica, sensilla styloconica, sensilla coeloconica, sensilla campaniformia, sensilla placodea, sensilla digitiformia, Böhm bristles, perforated plates, and cuticular pores. This is the first time that sensilla digitiformia has been reported in ladybirds. Finally, variations in mouthparts among ladybirds with differing diets, as well as the putative functions of each of the mouthparts and sensilla, were discussed. This research can provide a reference for understanding the functions of the mouthparts in ladybird feeding behavior and thereby contribute to the development of precise insect behavior regulation and management strategies.

Ultramorphology of the proboscis and sensilla of the Asian corn borer Ostrinia furnacalis (Lepidoptera: Crambidae).

The Asian corn borer, Ostrinia furnacalis (Guenée), is one of the most destructive lepidopteran pests affecting maize in Asia. Previous research on the feeding behavior of O. furnacalis has mostly focused on larvae, but the adult feeding preference has been unclear hitherto. In this study, we investigated the ultramorphology of the proboscis and sensilla of O. furnacalis using scanning electron microscopy and made morphological comparisons between the sexes, attempting to make predictions on the food preference and feeding behavior of the adults. The proboscis of O. furnacalis is divided into zones 1 and 2 with three types of sensilla, namely chaetica, basiconica, and styloconica. The entire proboscis and zone 2 of the females are significantly longer than those of the males. The main sexual difference in the sensilla is shown in the number of sensilla styloconica, which are eight in females but nine in males. The feeding preference of adult O. furnacalis is discussed based on the morphology of the proboscis and sensilla.

The earliest fossil mosquito.

Female mosquitoes are among the most notorious blood-feeding insects, sometimes causing severe allergic responses or vectoring a variety of microbial pathogens.1,2 Hematophagy in insects is likely a feeding shift from plant fluids, with the piercing-sucking mouthparts serving as suitable exaptation for piercing vertebrates' skin. The origins of these habits are mired in an often-poor fossil record for many hematophagous lineages,3,4 particularly those of sufficient age, as to give insights into the paleoecological context in which blood feeding first appeared or even to arrive at gross estimates as to when such shifts have occurred. This is certainly the case for mosquitoes, a clade estimated molecularly to date back to the Jurassic.5 The known Mesozoic Culicidae are Late Cretaceous, assigned to the modern Anophelinae or to the extinct Burmaculicinae, sister to other Culicidae, all with mouthparts of a modern type. Here, we report the discovery, in Lower Cretaceous amber from Lebanon, of two conspecific male mosquitoes unexpectedly with piercing mouthparts, armed with denticulate sharp mandibles and laciniae. These male fossils were likely hematophagous. They represent a lineage that diverged earlier than Burmaculicinae, extending the definitive occurrence of the family into the Early Cretaceous and serving to narrow the ghost-lineage gap for mosquitoes.

The genus Inversotyphlus Strasser, 1962, stat. nov. and Inversotyphlusammirandus sp. nov., a new bizarre, highly modified troglobiotic millipede (Diplopoda, Julida, Julidae) from Albania, Balkan Peninsula.

Inversotyphlus Strasser, 1962, stat. nov. is raised to the genus level and a new hydrophilous species, Inversotyphlusammirandussp. nov., is described from the second deepest pit in Albania. This species is characterized by a highly modified head and body for a presumably semiaquatic or hygropetric life and filtering diet. It is by far the most bizarrely modified cave-dwelling julid known. The new species is diagnosed, described in detail, and richly illustrated. Besides I.ammirandussp. nov., the genus Inversotyphlusstat. nov. includes six species: I.clavatus (Antic, 2018), comb. nov., I.edentulus (Attems, 1951), comb. nov., I.gellianae (Makarov & Rada, 2006), comb. nov., I.gracilis (Antic, 2018), comb. nov., I.lobifer (Attems, 1951), comb. nov., and I.opisthonodus (Antic, 2018) comb. nov. The subgenus Attemsotyphlussyn. nov. is here considered as a junior subjective synonym of the genus Inversotyphlusstat. nov. Notes are given on the habitat of I.ammirandussp. nov., the taxonomy of the tribe Typhloiulini and the genus Inversotyphlusstat. nov., and adaptive modifications of the mouthparts.

Microstructural Adaptation for Prey Manipulation in the Millipede Assassin Bugs (Hemiptera: Reduviidae: Ectrichodiinae).

Species in Ectrichodiinae are known for their prey specialization on millipedes. However, knowledge of the morphological adaptations to this unique feeding habit was limited. In the current study, we examined the microstructures of the antennae, mouthparts, and legs of four millipede feeding ectrichodiines, Ectrychotes andreae (Thunberg, 1888), Haematoloecha limbata Miller, 1953, Labidocoris pectoralis (Stål, 1863), and Neozirta eidmanni (Taueber, 1930), and compared them with those of three species of tribelocephalines, a group closely related to Ectrichodiinae. On the antennae, we found four types of antennal sensilla. On the mouthparts, we recognized four types of labial sensilla. Sampled ectrichodiines have distinctly more and denser slightly transverse ridges on the external side of mandibles than tribelocephalines. E. andreae and H. limbata possess numerous small papillae fringed with densely arranged finger-print-like grains on the trochanter and femur; these probably facilitate the immobilization of prey. Overall, our study illustrates, at a microstructural level, the remarkable morphological adaption of prey manipulation in ectrichodiine, and has enhanced our understanding about stenophagy in the family Reduviidae.

Honey bees switch mechanisms to drink deep nectar efficiently.

The feeding mechanisms of animals constrain the spectrum of resources that they can exploit profitably. For floral nectar eaters, both corolla depth and nectar properties have marked influence on foraging choices. We report the multiple strategies used by honey bees to efficiently extract nectar at the range of sugar concentrations and corolla depths they face in nature. Honey bees can collect nectar by dipping their hairy tongues or capillary loading when lapping it, or they can attach the tongue to the wall of long corollas and directly suck the nectar along the tongue sides. The honey bee feeding apparatus is unveiled as a multifunctional tool that can switch between lapping and sucking nectar according to the instantaneous ingesting efficiency, which is determined by the interplay of nectar-mouth distance and sugar concentration. These versatile feeding mechanisms allow honey bees to extract nectar efficiently from a wider range of floral resources than previously appreciated and endow them with remarkable adaptability to diverse foraging environments.

The Mouthparts of Female Blood-Feeding Frog-Biting Midges (Corethrellidae, Diptera).

Females of frog-biting midges (Corethrellidae) obtain their blood meals from male calling frogs. While the morphology of the feeding apparatus is well studied in hematophagous Diptera that impact humans, frog-biting midges have received far less attention. We provide a detailed micromorphological examination of the piercing blood-sucking proboscis and maxillary palpus in three Corethrella species using scanning electron microscopy and histological semi-thin sectioning. We also compare the sensilla found on the proboscis tip and the palpus of Corethrella with other piercing blood-sucking Diptera. Corethrella spp. have a proboscis length of about 135 µm, equipped with delicate mandibular piercing structures composing the food canal together with the labrum and hypopharynx. Their proboscis composition is plesiomorphic and more similar to other short-proboscid hematophagous Culicomorpha (e.g., Simuliidae), in contrast to the phylogenetically more closely related long-proboscid Culicidae. As in other short-proboscid taxa, the salivary canal in Corethrella spp. transitions into an open salivary groove with one mandible forming a seal, whereas in Culicidae the salivary canal is closed until the tip of the proboscis. We discuss the possible functional constraints of very short, piercing blood-sucking proboscises (e.g., dimensions of host blood cells) that may limit the size of the food canal.

Simultaneous Detection and Distribution of Five Juvenile Hormones in 58 Insect Species and the Absolute Configuration in 32 Insect Species.

Juvenile hormone (JH) plays an important role in regulating various insect physiological processes. Herein, a novel method (chiral and achiral) for the simultaneous detection of five JHs was established by processing a whole insect without complicated hemolymph extraction. The proposed method was used to determine the distribution of JHs in 58 insect species and the absolute configuration of JHs in 32 species. The results showed that JHSB3 was uniquely synthesized in Hemiptera, JHB3 was unique to Diptera, and JH I and JH II were unique to Lepidoptera. JH III was present in most insect species surveyed, with social insects having generally higher JH III titers. Interestingly, JHSB3 and JHB3, both double epoxidation JHs, were found in insects with sucking mouthparts. The absolute conformation of JH III and the 10C of the detected JHs were all R stereoisomers.

A switchable joint in the head of dragonfly larvae (Insecta: Odonata) as key to the multifunctionality of the prehensile labial mask.

Dragonfly and damselfly larvae (Insecta: Odonata) capture prey by rapid protraction of a raptorial mouthpart, based on a modified labium. Yet, in insects with biting-chewing mouthparts, the labium has an essential role in food handling. These two distinct functions -prey capturing and handling-lead to a mechanical problem in Odonata larvae: while the labium is always protracted in a straight line during prey capture, food handling requires more dexterity. In this study, we investigate the role of the labium in the feeding process and analyse the mechanics of the labial joints in the dragonfly larva Anax imperator. Our results show that the labium features a multiaxial joint connecting the basal segment (postmentum) and the head. During feeding, a combination of rotations around different axes is used to handle and orient prey, which is unique among biting-chewing mouthparts. Furthermore, we identified structures at the joint which likely restrict lateral motion during the predatory strike. Our results provide a further understanding of the unique prey-capturing apparatus of odonate larvae capable of controlling a 'switchable' multiaxial to a restricted monoaxial joint. This concept highlights the evolution of a highly modified raptorial mouthpart appendage where the degrees of freedom can be actively restricted to allow for the respectively needed functionality.

Scanning Electron Microscopy of Antennae and Mouthparts of Mezira yunnana Hsiao (Hemiptera: Aradidae): Specialized Microstructures Reflecting Adaptation to Mycetophagy.

Many species of the family Aradidae (also known as flat bugs) feed on fungal mycelia and fruiting bodies. In order to better understand the morphological adaptation to this unique feeding habit, we examined the microstructure of antennae and mouthparts of an aradid species, Mezira yunnana Hsiao, using scanning electron microscope, and documented the fungal feeding process under laboratory conditions. The antennal sensilla include three subtypes of sensilla trichodea, three subtypes of sensilla basiconica, two subtypes of sensilla chaetica, sensilla campaniformia, and sensilla styloconica. The apex of the second segment of flagellum has a large number of various sensilla forming a sensilla cluster. The labial tip is distally constricted, which is rarely observed in other Pentatomomorpha species. The labial sensilla include three subtypes of sensilla trichodea, three subtypes of sensilla basiconica, and a sensilla campaniformia. The tip of the labium has only three pairs of sensilla basiconica III and small comb-shaped cuticular processes. The external surface of the mandibular apex has 8-10 ridge-like central teeth. A series of key morphological structures associated with mycetophagous feeding habit were identified, which will facilitate future studies on adaptive evolution of species in Pentatomomorpha as well as in other heteropteran lineages.

Aphagy and vestigial stylets in first-instar nymphs of Aradidae (Hemiptera, Heteroptera).

Aradidae are known for their remarkably long stylets, coiled at rest in the anterior part of the head. However, previous reports indicated that at least some species lacked stylets during the first nymphal instar. A more detailed examination of Aradus betulae 1st-instar nymphs showed that their mandibular and maxillary stylets are abnormally short, not coiled, improperly interlocked, and clearly non-functional. The anteclypeus is relatively small and its internal diverticulum, which accommodates the stylet coil in the older stages, is vestigial. In contrast, the labium, labrum, food canal, and associated structures and muscles, including protractors and retractors of the stylets, are all normally developed. First-instar nymphs of Aradidae are the first known Heteroptera with non-functional mouthparts. To explain this phenomenon, a hypothesis is proposed which links previously unexplained records of non-feeding (but endowed with regular, functional mouthparts) 1st-instar nymphs of various pentatomomorphan families with the special role of that stage in acquiring microbial gut symbionts. A presumed loss of symbionts in the ancestors of Aradidae may have led to reduction of the now useless stylets in the first instar, which retained aphagy, despite a spectacular elongation of stylets in the older, feeding instars.

The mouthparts of the Aradidae (Insecta: Hemiptera: Heteroptera).

The flat bugs, Aradidae, have exceptionally long piercing-sucking stylets coiled up at rest in the anterior part of the head. Previous studies suggested that the majority of aradids can be divided into two groups by the direction of stylet coiling, clockwise or anticlockwise. Detailed reconstruction of the head skeleton and musculature from series of polished sections, examined in SEM, of epon-embedded specimens of three species has shown that these groups represent two disparate modifications of the head groundplan. In Aradus betulae (L.), the stylet coil is accommodated inside the greatly enlarged anteclypeus within an expansible membranous diverticulum of its epipharyngeal cuticle. In contrast, in Isodermus planus Erichson and Carventus brachypterus Kormilev, the coil lies freely underneath the anteclypeus between the extended maxillary lobes (in I. planus fused with the extended gular lobe). The intraclypeal coils occur in the subfamilies Aradinae, Calisiinae, and Chinamyersiinae and the subclypeal coils in Isoderminae, Carventinae, Mezirinae, Aneurinae, Prosympiestinae, and possibly in the closely related family Termitaphididae. Each method of stylet coiling is associated with a suite of divergently specialized structural traits, suggesting that the two groups have independently evolved from ancestors endowed with regular stylets. Functional mechanics of the coiled stylet bundles are discussed.

Evolutionary functional morphology of the proboscis and feeding apparatus of hawk moths (Sphingidae: Lepidoptera).

The morphology of the proboscis and associated feeding organs was studied in several nectar-feeding hawk moths, as well as a specialized honey-feeder and two supposedly nonfeeding species. The proboscis lengths ranged from a few millimeters to more than 200 mm. Despite the variation in proboscis length and feeding strategy, the principle external and internal composition of the galeae, the stipes pump, and the suction pump were similar across all species. The morphology of the smooth and slender proboscis is highly conserved among all lineages of nectar-feeding Sphingidae. Remarkably, they share a typical arrangement of the sensilla at the tip. The number and length of sensilla styloconica are independent from proboscis length. A unique proboscis morphology was found in the honey-feeding species Acherontia atropos. Here, the distinctly pointed apex displays a large subterminal opening of the food canal, and thus characterizes a novel type of piercing proboscis in Lepidoptera. In the probably nonfeeding species, the rudimentary galeae are not interlocked and the apex lacks sensilla styloconica; galeal muscles, however, are present. All studied species demonstrate an identical anatomy of the stipes, and suction pump, regardless of proboscis length and diet. Even supposedly nonfeeding Sphingidae possess all organs of the feeding apparatus, suggesting that their proboscis rudiments might still be functional. The morphometric analyses indicate significant positive correlations between galea lumen volume and stipes muscle volume as well as the volume of the food canal and the muscular volume of the suction pump. Size correlations of these functionally connected organs reflect morphological fine-tuning in the evolution of proboscis length and function.

Mouthpart Ecomorphology and Predatory Behaviour in Selected Rove Beetles of the "Staphylinine Group" (Coleoptera: Staphylinidae: Staphylininae, Paederinae).

The representatives of the megadiverse rove beetle subfamilies Paederinae and Staphylininae (Coleoptera: Staphylinidae) are considered generalist predators, although their exact prey-capture behaviour and performance and possible links to mouthpart morphology have rarely been described. Here, we examine these relationships for selected species by SEM analyses of mouthparts and front legs and highspeed videography of prey-capture behaviour. We describe the observed behaviours and structural properties and quantify relationships between prey type, mouthpart morphology, and predatory performance based on morphometric measurements of both the shape and lever properties of the mandible. We show that the Staphylininae considered have morphological and behavioural properties generally associated with generalist predation and that the Paederinae considered display characteristics that are highly specialized on elusive prey such as Collembola. We found correlations between mandible shape and leverage, and body size and prey type. We report distinct prey-capture behaviours: the beetles use front legs and/or mandibles to attack prey, drag prey, or cage it between their legs. These strategies differ among species and situations. Overall, this exploratory study provides insights into the morphology and types of prey capture that must have played a major role in the evolution of these beetles.

Concrete Urban Refugia for Culex quinquefasciatus, Impact of Summer Stressors, and Strategies Adopted to Oversummer.

We carried out vector surveillance to locate adult sites of Culex quinquefasciatus, a filarial vector, during the summer in North Campus, Delhi University, Delhi, India. It revealed that the adults were a rare sight but landed naturally and assembled in indoor sites during the peak summer in 2016 and 2019. New adults repopulated some of these sites, after the removal of the samples. These adults were used to study the impact of summer stressors on their morphological, physiological, and behavioral traits, and strategies followed to oversummer in the urban environment. Adults adopted endophily, vagility, and staying at a lower height as strategies to avoid lethality. Females outnumbered the males, and showed morphological, physiological, and behavioral consequences; the majority were unfed. In females, sublethal effects such as aberrations of body parts including mouthparts; distorted oviposition pattern, reduced fecundity, and precocious tanning of retained eggs; and reduced egg hatch rate contributed to reduced survival and reduced reproductive output, thereby reducing vector load during peak summer. These are novel findings. Source reduction of adults during this period would provide effective eco-friendly control of this established species and should be part of the vector management strategies so that swarms of adults during monsoon could be prevented.

Mouthpart structure of the adult Bicaubittacus appendiculatus () (Mecoptera: Bittacidae).

The structure and functional morphology of the mouthparts were investigated in adult hangingfly Bicaubittacus appendiculatus (Esben-Petersen, 1927) by scanning electron microscopy and histological serial sections. The mandibulate mouthparts consist of a labrum-epipharynx, paired mandibles and maxillae, and unpaired labium and hypopharynx. The labrum is elongated and tapered toward the apex. The epipharynx is furnished with numerous sensilla. The mandibles are sword-shaped, with an outer sharp tooth curved mesad and an inner blunt corner. The basal region of each mandible processes a conical projection. The maxillae are well-developed, each consisting of a sclerotized cardo, an elongated stipes, which bears an inner lacinia, an outer galea, and laterally a five-segmented maxillary palp. The labium is formed by a postmentum, a prementum and a pair of two-segmented labial palps. The hypopharynx is concave inward on the anterior side, with numerous microtrichia on the posterior surface. Seven types of sensilla were found on the mouthparts: sensilla basiconica on the epipharynx, and maxillary and labial palps; sensilla chaetica on the epipharynx; sensilla palmata, sensilla placoidea and sensilla trichodea on the epipharynx and maxillary palp; sensilla campaniformia and hair plates on the basal joints of palps. The sensillar function and the feeding mechanism of mouthparts in bittacids are briefly discussed.

Possible long-proboscid insect pollinators from the Early Permian of Russia.

Insect pollination is one of the hallmarks of flowering plants.1 Bees, moths, flies, and some other pollinators evolved elongate siphonate mouthparts for sucking concealed nectar and occasionally other liquids.2 However, it is clear from the fossil record that insects with similar adaptations appeared long before the mid-Cretaceous radiation of angiosperms. These insects most probably used their proboscis to reach pollination drops and other sugary fluids that were hidden in the cones of extinct gymnosperms, pollinating them in the process.3-6 The vast majority of these gymnosperm-associated long-proboscid insects have been reported from the Middle Jurassic to the Early Cretaceous, i.e., the time interval that immediately predated the advent of flowering plants.7 By contrast, the Paleozoic stage of the co-evolution between long-proboscid insect pollinators and plants has remained poorly understood. Here, we report a putative pollination mutualism involving long-proboscid holometabolous insects (Panorpida: Protomeropidae) from the Early Permian of Russia (ca. 283-273 Ma). Their elongate mouthparts have very similar morphology to those of some present-day nectarivorous Coleoptera and Hymenoptera and probably served to imbibe micropylar secretions from the semi-closed ovulate organs of the gymnosperms of a peltaspermalean affinity that have been found in the same locality. This is the earliest record of insects with siphonate-like mouthparts, which could indicate that the complex interactions between pollinators and gymnosperms predate the first flowering plants by over 100 Ma.

"Natural infections" with Trypanosoma cruzi via the skin of mice: size of mouthparts of vectors and numbers of invading parasites.

Investigating parameters influencing natural infections with Trypanosoma cruzi via the skin, the diameters of mouthparts of different stages of triatomines vectors were measured to determine the size of the channel accessible for T. cruzi during cutaneous infection. The mean diameters of the skin-penetrating mandibles of first to fifth instar nymphs of the vector Triatoma infestans increased from 18 to 65 µm. The mean diameter in fourth instar nymphs of Dipetalogaster maxima was 86 µm. Different numbers of isolated vector-derived metacyclic trypomastigotes (10-10,000) were injected intradermally into mice. Prepatent periods, parasitemia and mortality rates were compared with those of mice obtaining 10,000 metacyclic trypomastigotes that are usually present in the first drop of faeces onto the feeding wounds of fifth and fourth instar nymphs of T. infestans and D. maxima, respectively. After injection of 50-10,000 T. cruzi, in all 42 mice the infection developed. An injection of 10 parasites induced an infection in 8 out of 15 mice. With increasing doses of parasites, prepatent periods tended to decrease. The level of parasitemia was higher after injection of the lowest dose. Except for one mouse all infected mice died. After placement of 10,000 metacyclic trypomastigotes onto the feeding wound of fifth or fourth instar nymphs of T. infestans and D. maxima, respectively, the infection rates of the groups, prepatent periods and the levels of parasitemia of T. cruzi in mice indicated that about 10-1,000 metacyclic trypomastigotes entered the skin via this route. For the first time, the present data emphasise the risk of an infection by infectious excreta of triatomines deposited near the feeding wound and the low number of invading parasites.