Titanium and iron titanium oxide nanoparticles in antennae of the migratory ant Pachycondyla marginata: an alternative magnetic sensor for magnetoreception?

The most accepted hypothesis of magnetoreception for social insects is the ferromagnetic hypothesis which assumes the presence of magnetic material as a sensor coupled to sensitive structures that transmit the geomagnetic field information to the nervous system. As magnetite is the most common magnetic material observed in living beings, it has been suggested as basic constituent of the magnetoreception system. Antennae and head have been pointed as possible magnetosensor organs in social insects as ants, bees and termites. Samples of three antenna joints: head-scape, scape-pedicel and pedicel-third segment joints were embedded in epoxi resin, ultrathin sectioned and analyzed by transmission electron microscopy. Selected area electron diffraction patterns and X-ray energy dispersive spectroscopy were obtained to identify the nanoparticle compound. Besides iron oxides, for the first time, nanoparticles containing titanium have been identified surrounded by tissue in the antennae of ants. Given their dimension and related magnetic characteristics, these nanoparticles are discussed as being part of the magnetosensor system.

Differential scaling within an insect compound eye.

Environmental and genetic influences cause individuals of a species to differ in size. As they do so, organ size and shape are scaled to available resources whilst maintaining function. The scaling of entire organs has been investigated extensively but scaling within organs remains poorly understood. By making use of the structure of the insect compound eye, we show that different regions of an organ can respond differentially to changes in body size. Wood ant (Formica rufa) compound eyes contain facets of different diameters in different regions. When the animal body size changes, lens diameters from different regions can increase or decrease in size either at the same rate (a 'grade' shift) or at different rates (a 'slope' shift). These options are not mutually exclusive, and we demonstrate that both types of scaling apply to different regions of the same eye. This demonstrates that different regions within a single organ can use different rules to govern their scaling, responding differently to their developmental environment. Thus, the control of scaling is more nuanced than previously appreciated, diverse responses occurring even among homologous cells within a single organ. Such fine control provides a rich substrate for the diversification of organ morphology.

New exocrine glands in ants: the hypostomal gland and basitarsal gland in the genus Melissotarsus (Hymenoptera: Formicidae).

Fisher and Robertson (Insect Soc 46: 78-83, 1999) discovered the production of silk-like secretions emerging from slit-shaped openings along the anterior margin of the ventral hypostoma of Melissotarsus ant workers. The current histological study describes a hitherto unknown hypostomal gland from which this silk-like substance originates. In addition, this study describes a new basitarsal gland in the three pairs of legs of Melissotarsus workers.

Description of Cephalotes specularis n. sp. (Formicidae: Myrmicinae)-the mirror turtle ant.

We describe here Cephalotes specularis n. sp. (Hymenoptera: Formicidae: Myrmicinae: Cephalotini) based on minor and major workers, gynes and larvae from Uberlândia, Minas Gerais state, Brazil. On morphological and molecular grounds, this new species belongs to the C. fiebrigi + C. bruchi species complex, of which there are 11 previously described species (one in C. bruchi group and 10 in the C. fiebrigi group). All members of these groups are found in, or are limited to the South American "arid diagonal", comprised of the Argentinian Chaco, the Cerrados of central South America, and the Brazilian northeastern caatingas. Workers of C. specularis n. sp. have an extremely shiny gaster which is mirror-like, notwithstanding its sparse covering by minute hairs. This species engages in a form of resource-based social parasitism of the host ant Crematogaster ampla (Myrmicinae: Crematogastrini). Cephalotes specularis foragers move freely in the dense traffic of Crematogaster ampla foraging trails. They exhibit highly atypical body posturing for turtle ants, which makes them hard to distinguish from the Crematogaster foragers.

Scanning ultrafast electron microscopy.

Progress has been made in the development of four-dimensional ultrafast electron microscopy, which enables space-time imaging of structural dynamics in the condensed phase. In ultrafast electron microscopy, the electrons are accelerated, typically to 200 keV, and the microscope operates in the transmission mode. Here, we report the development of scanning ultrafast electron microscopy using a field-emission-source configuration. Scanning of pulses is made in the single-electron mode, for which the pulse contains at most one or a few electrons, thus achieving imaging without the space-charge effect between electrons, and still in ten(s) of seconds. For imaging, the secondary electrons from surface structures are detected, as demonstrated here for material surfaces and biological specimens. By recording backscattered electrons, diffraction patterns from single crystals were also obtained. Scanning pulsed-electron microscopy with the acquired spatiotemporal resolutions, and its efficient heat-dissipation feature, is now poised to provide in situ 4D imaging and with environmental capability.

On the morphology of the digestive system of two Monomorium ant species.

The digestive system of adults and mature larvae of two ant species of Monomorium Mayr (Hymoneptera: Formicidae) were described with the aid of light and scanning electron microscopy, as there is a lack of studies in this area. These two ant species are recurrently found in urban habitats and are known as 'tramp species,' as they cause problems in households, businesses, and hospitals. The most interesting finds of the present study include the existence of spinules in the crop of adults, and the number of Malpighian tubules and rectal pads was constant among different castes, ages, and species.

The influence of surface energy on the self-cleaning of insect adhesive devices.

The ability of insects to adhere to surfaces is facilitated by the use of adhesive organs found on the terminal leg segments. These adhesive pads are inherently 'tacky' and are expected to be subject to contamination by particulates, leading to loss of function. Here, we investigated the self-cleaning of ants and beetles by comparing the abilities of both hairy and smooth pad forms to self-clean on both high and low energy surfaces after being fouled with microspheres of two sizes and surface energies. We focused on the time taken to regain adhesive potential in unrestrained Hymenopterans (Polyrhachis dives and Myrmica scabrinodis) and Coccinellids (Harmonia axyridis and Adalia bipunctata) fouled with microspheres. We found that the reattainment of adhesion is influenced by particle type and size in Hymenopterans, with an interaction between the surface energy of the contaminating particle and substrate. In Coccinellids, reattainment of adhesion was only influenced by particle size and substrate properties. The adhesive organs of Coccinellids appear to possess superior self-cleaning abilities compared with those of Hymenopterans, although Hymenopterans exhibit better adhesion to both surface types.

Compared morphology of the immatures of males of two urban ant species of Camponotus.

The immatures of males of two species of Camponotus ants (Hymenoptera: Formicidae) are described and compared by light and electron microscopy. The numbers of larval instars were determined: Camponotus rufipes Fabricius (Hymenoptera: Formicidae) have four instars; and Camponotus vittatus Forel have three. Male larvae of the two species are similar to previously described Camponotus larvae, sharing the following traits: basic shape of body and mandible, presence of 'chiloscleres', 'praesaepium' (some specimens), labial pseudopalps, and ten pairs of spiracles. However, larvae of the two species can be separated by bodily dimensions and based on their hair number and types. Worker larvae of C. vittatus previously described are extensively similar to male larvae, with only a few inconspicuous differences that may result from intraspecific variation or sexual differences.

Description of the immatures of the ant, Myrmelachista catharinae.

The Neotropical ant genus Myrmelachista Roger comprises 69 described species and subspecies, and still is a poorly studied group. Larvae play a paramount role in colony nutrition in social hymenopterans and bear considerable value in the reconstruction of group phylogenies, however, they are generally neglected. Larvae of different instars of Myrmelachista catharinae Mayr (Hymenoptera: Formicidae) are herein described in detail by light and scanning electron microscopy. The number of larval instars was estimated as three based on the frequency distribution of maximum head capsule widths. The described larvae confirmed some traits typical of the genus: general shape of body and mandibles, general aspect and distribution of body hairs, and the number of sensilla on the palps and galea. Differently from other Myrmelachista larvae previously described, M. catharinae presented two distinct kinds of second instars, some additional types of body hairs, different number of antennal sensilla, and a distinct labrum shape. M. catharinae presented ten pairs of spiracles, which is the first record for this genus.

First record of the myrmicine ant genus Syllophopsis Santschi, 1915 (Hymenoptera: Formicidae) from India with description of a new species.

Here we describe and illustrate Syllophopsis peetersi sp. nov. from Silent Valley National Park, a biodiversity hotspot region of the Western Ghats of India. The discovery also marks a first native report of the genus from the Indian subcontinent. Scanning Electron Microscopy (SEM) analysis was carried out to elucidate the general morphology and sensilla of the new species. The new species is similar to congeners from Madagascar, but with larger differences from species that occur elsewhere.

The basitarsal sulcus gland, a novel exocrine structure in ants.

The basitarsus of the mid- and/or hindlegs of several Amblyoponinae ants shows a deep longitudinal groove or sulcus on its anterior face in workers and queens. Histological examination reveals this sulcus is associated with a conspicuous novel epithelial gland, which brings the number of exocrine glands in the legs of ants to 25. The ultrastructural characteristics of the gland show the presence of a well-developed smooth endoplasmic reticulum. This is indicative for the elaboration of a non-proteinaceous and thus possibly pheromonal secretion. Behavioural observations show that this secretion is collected by the tarsomeres and spread onto the brood and nest, suggesting a role in nestmate recognition. A similar basitarsal sulcus gland was also found in Nothomyrmecia, Paraponera and Tetraponera, which represents both a wide phylogenetic and ecological distribution, as it includes arboreal, ground-dwelling as well as subterranean taxa.

Description of the immatures of Linepithema humile Mayr (Hymenoptera: Formicidae).

Linepithema humile Mayr is an ant species originally native to South America that has been spread accidently throughout the globe through international trade. It is a serious urban and crop pest. Despite its economic importance, little is known about the larvae of this species apart from a brief description based on a few specimens. The present investigation is aimed at describing every immature stage of L. humile. Three larval instars were determined through the frequency distribution of the maximum width of head capsules from a sample of 525 larvae. The morphological descriptions were based on 150 eggs, 70 larvae, and 90 pupae examined by light and scanning electron microscopy. Some morphological characteristics reported to be typical of Linepithema Mayr larvae were confirmed--dolichoderoid body shape, presence of dorsal protuberance, sparse simple body hairs, presence of nine pairs of spiracles and dolichoderoid mandibles. We concluded that an earlier published description was based on queen larvae, and that the protuberance is only present in worker larvae. The information provided in this study may aid ant systematics and phylogenetics, as well provide a better understanding of the biology of this species.

Toxicological and histopathological effects of boric acid on Atta sexdens rubropilosa (Hymenoptera: Formicidae) workers.

The current study compared the toxicity of different concentrations of boric acid in adult workers of Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae), with toxicological bioassays, and examining the dose-dependent and time-dependent histopathological changes, of the midgut, Malpighian tubules, and postpharyngeal glands. Our results revealed the importance of conducting toxicological bioassays combined with morphological analyses of the organs of ants chronically exposed to insecticides used in commercial ant baits. In vitro bioassays showed that boric acid significantly decreases the survivorship of workers regardless of concentration, whereas the morphological data suggested progressive dose-dependent and time-dependent changes in the organs examined, which were evident in the midgut. The midgut is the first organ to be affected, followed by the postpharyngeal gland and Malpighian tubules. This sequence is in agreement with the absorption pathway of this chemical compound in the midgut, its transference to the hemolymph, possibly reaching the postpharyngeal glands, and excretion by the Malpighian tubules. These progressive changes might be due to the cumulative and delayed effect of boric acid. Our findings provide important information for the understanding of the action of boric acid in ant baits in direct and indirect target organs.

General morphology and ultrastructure of the venom apparatus and convoluted gland of the fire ant, Solenopsis saevissima.

A group of 13 species of the genus Solenopsis is markedly difficult to assess taxonomically, although they are of considerable economical and medical importance in some countries where some of them were introduced. These ants are aggressive and their venomous stings can be very allergenic. The venom apparatus has been described in fine detail for only two of these species, and differences in this structure among the different species might prove useful as taxonomic characters. The venom apparatus of Solenopsis saevissima Smith (Hymenoptera: Formicidae) is herein described with the aid of light and electron microscopy techniques, and compared to that of S. invicta and S. richteri. The cellular organization of the different parts present differences that suggest functional specialization. In general, the different tissues were abundant in vesiculae and mitochondria, but presented little endoplasmic reticulum and few ribosomes, probably because they produce little protein. The length of the free filaments of the venom gland and the width of their internal ducts seems to vary from what was described for S. richteri, but this may be of little use to taxonomy.

Morphological description of the immatures of the ant, Monomorium floricola.

Some ant species of the genus Monomorium Mayr occur worldwide and are considered important urban pests. The larvae of only a few species of this genus have been described, and these descriptions are either superficial or incomplete. This study aimed to determine the number of larval instars and describe the immature stages of the ant Monomorium floricola Jerdon (Formicidae: Myrmicinae). Specimens were analyzed and measured using light and scanning electron microscopy. Three larval instars were found, and all larvae had pheidoloid bodies with ectatommoid mandibles, consistent with other Monomorium species described previously. Five types of body hairs were described, and their distribution was instar-specific. Body and mandible dimensions of the larvae also were constant for each instar. Like other Myrmicinae, the larvae did not create a cocoon. Some of differences among the hair types and sensilla were observed by comparing the samples with larvae of other species in the genus, and these differences may have taxonomic utility.

Novel exocrine glands in the foreleg coxae of Discothyrea ants.

Workers, queens and males of all examined Discothyrea species of the 'sauteri group', that have laterally expanded frontal lobes and well-developed antennal scrobes, are characterized by two hairy areas on the outer surface of their procoxae. Histological and ultrastructural examination of Discothyrea sauterirevealed that each of these areas is associated with a novel exocrine gland: the proximal procoxal gland is formed by a cluster of 15 round secretory cells of 34 µm with numerous mitochondria, smooth endoplasmic reticulum and Golgi apparatus.Their ducts have a diameter of 0.5-1 µm.The distal procoxal gland contains 50 secretory cells of 22 µm with numerous vacuoles and lamellar inclusions, and narrow ducts with a diameter of only 0.15-0.2 µm.The differences in ultrastructural appearance and duct diameter indicate that both glands produce a different but probably pheromonal secretion. The function of these novel procoxal glands could not yet be determined, although observation of D.sauteri workers and queens shows that they make frequent and peculiar leg movements, in which the foreleg basitarsus rubs over the coxal hairy areas. The foreleg basitarsus then rubs the ipsilateral hindleg basitarsus and antenna. As a last step of the sequence, the hindleg basitarsus strokes the gaster.In addition to the occurrence of these novel procoxal glands, histological examination of D. sauteri also revealed the presence of yet another novel but smaller procoxal base gland. Ants of the 'testacea group', that have less developed frontal lobes and no antennal scrobes, do not have procoxal hairy areas, although a distinct sculpturation with small pores may occur in the corresponding areas. The related Proceratium japonicum, that has a similar lifestyle as Discothyrea, does not have any of the procoxal glands and does not display the peculiar leg movements as reported for D. sauteri.

Glandular innovations for a tunnelling life: Silk and associated leg glands in Melissotarsus and Rhopalomastix queen and worker ants.

As in other Hymenoptera, adult ants cannot secrete silk, unlike the larvae that spin a cocoon prior to metamorphosis. Fisher and Robertson (1999) first showed the existence of a silk gland in the head of adult Melissotarsus beccarii workers, and we confirm this with detailed histology and ultrastructural comparisons of both queens and workers. This African genus exhibits extreme morphological adaptations (legs, head shape and mandibular muscles) for tunnelling behaviour inside living trees, that underlie an obligate mutualism with scale insects. Rhopalomastix is its sister genus distributed across Asia, and we show that queens and workers also have a silk gland. This lineage of minute workers relies on silk to secure their network of tunnels against other arboreal ants. We show striking differences between these genera in the anatomy and ultrastructure of the cells that secrete silk, especially numerous vacuoles and an unexpectedly branched end apparatus in Melissotarsus. Moreover, the legs of Melissotarsus are much more specialized for tunnelling, and this includes highly expanded basitarsi. The latter house the novel 'Delage-Darchen gland', and we document its anatomy and ultrastructure, suggesting a proteinaceous secretion to harden roofs made of silk combined with wood fragments. The restriction of the Delage-Darchen gland to Melissotarsus, combined with a modified silk gland (an almost three-fold increase in the number of secretory cells, and ultrastructural differences suggestive of higher secretory activity), are evidence of an outstanding evolutionary divergence relative to Rhopalomastix. Synthesis of silk by adults is a significant innovation among ants, but its augmented production in Melissotarsus makes them better adapted for the hazards of arboreal life.

Distal leg structures of the Aculeata (Hymenoptera): A comparative evolutionary study of Sceliphron (Sphecidae) and Formica (Formicidae).

The distal parts of the legs of Sceliphron caementarium (Sphecidae) and Formica rufa (Formicidae) are documented and discussed with respect to phylogenetic and functional aspects. The prolegs of Hymenoptera offer an array of evolutionary novelties, mainly linked with two functional syndromes, walking efficiently on different substrates and cleaning the body surface. The protibial-probasitarsomeral cleaning device is almost always well-developed. A complex evolutionary innovation is a triple set of tarsal and pretarsal attachment devices, including tarsal plantulae, probasitarsomeral spatulate setae, and an arolium with an internal spring-like arcus, a dorsal manubrium, and a ventral planta. The probasitarsal adhesive sole and a complex arolium are almost always preserved, whereas the plantulae are often missing. Sceliphron has retained most hymenopteran ground plan features of the legs, and also Formica, even though the adhesive apparatus of Formicidae shows some modifications, likely linked to ground-oriented habits of most ants. Plantulae are always absent in extant ants, and the arolium is often reduced in size, and sometimes vestigial. The arolium contains resilin in both examined species. Additionally, resilin enriched regions are also present in the antenna cleaners of both species, although they differ in which of the involved structures is more flexible, the calcar in Sceliphron and the basitarsal comb in Formica. Functionally, the hymenopteran distal leg combines (a) interlocking mechanisms (claws, spine-like setae) and (b) adhesion mechanisms (plantulae, arolium). On rough substrate, claws and spine-like setae interlock with asperities and secure a firm grip, whereas the unfolding arolium generates adhesive contact on smooth surfaces. Differences of the folded arolium of Sceliphron and Formica probably correlate with differences in the mechanism of folding/unfolding.

Sex-specific antennal sensory system in the ant Camponotus japonicus: structure and distribution of sensilla on the flagellum.

The antennae are a critically important component of the ant's highly elaborated chemical communication systems. However, our understanding of the organization of the sensory systems on the antennae of ants, from peripheral receptors to central and output systems, is poorly understood. Consequently, we have used scanning electron and confocal laser microscopy to create virtually complete maps of the structure, numbers of sensory neurons, and distribution patterns of all types of external sensilla on the antennal flagellum of all types of colony members of the carpenter ant Camponotus japonicus. Based on the outer cuticular structures, the sensilla have been classified into seven types: coelocapitular, coeloconic, ampullaceal, basiconic, trichoid-I, trichoid-II, and chaetic sensilla. Retrograde staining of antennal nerves has enabled us to count the number of sensory neurons housed in the different types of sensilla: three in a coelocapitular sensillum, three in a coeloconic sensillum, one in an ampullaceal sensillum, over 130 in a basiconic sensillum, 50-60 in a trichoid-I sensillum, and 8-9 in a trichoid-II sensillum. The basiconic sensilla, which are cuticular hydrocarbon-receptive in the ant, are present in workers and unmated queens but absent in males. Coelocapitular sensilla (putatively hygro- and thermoreceptive) have been newly identified in this study. Coelocapitular, coeloconic, and ampullaceal sensilla form clusters and show biased distributions on flagellar segments of antennae in all colony members. The total numbers of sensilla per flagellum are about 9000 in unmated queens, 7500 in workers, and 6000 in males. This is the first report presenting comprehensive sensillar maps of antennae in ants.

First report of endocytobionts in the digestive tract of ponerine ants.

We conducted ultrastructural studies to examine the presence of microorganisms in the midgut of the ant Odontomachus bauri (Ponerinae), as a contribution towards understanding the relationships between microorganisms and their hosts. The presence of microorganisms in this region, including inside the cells, suggests their participation in food digestion as symbiontic organisms and represent a new possibility to exploit food sources in the environment.