Parasitoid attachment ability and the host surface wettability.

Climbing animals such as geckos and arthropods developed astonishing adhesive mechanisms which are fundamental for their survival and represent valuable models for biomimetic purposes. A firm adhesion to the host surface, in order to successfully lay eggs is necessary for the reproduction of most parasitoid insects. In the present study, we performed a comparative investigation on the attachment ability of four parasitoid species (the egg parasitoid Anastatus bifasciatus (Eupelmidae), the aphid parasitoid Aphidius ervi (Braconidae), the fly pupal ectoparasitoid Muscidifurax raptorellus (Pteromalidae) and the pupal parasitoid of Drosophila Trichopria drosophilae (Diapriidae)) with hosts characterized by a surface having different wettability properties. The friction force measurements were performed on smooth artificial (glass) surfaces showing different contact angles of water. We found that attachment systems of parasitoid insects are tuned to match the wettability of the host surface. Sexual dimorphism in the attachment ability of some tested species has been also observed. The obtained results are probably related to different microstructure and chemical composition of the host surfaces and to different chemical composition of the parasitoid adhesive fluid. The data here presented can be interpreted as an adaptation, especially in the female, to the physicochemical properties of the host surface and contribute to shed light on the coevolutionary processes of parasitoid insects and their hosts.

Insect attachment on waxy plant surfaces: the effect of pad contamination by different waxes.

This study focuses on experimental testing of the contamination hypothesis and examines how the contamination of insect adhesive pads with three-dimensional epicuticular waxes of different plant species contributes to the reduction of insect attachment. We measured traction forces of tethered Chrysolina fastuosa male beetles having hairy adhesive pads on nine wax-bearing plant surfaces differing in both shape and dimensions of the wax structures and examined insect adhesive organs after they have contacted waxy substrates. For comparison, we performed the experiments with the same beetle individuals on a clean glass sample just before (gl1) and immediately after (gl2) the test on a plant surface. The tested insects showed a strong reduction of the maximum traction force on all waxy plant surfaces compared to the reference experiment on glass (gl1). After beetles have walked on waxy plant substrates, their adhesive pads were contaminated with wax material, however, to different extents depending on the plant species. The insects demonstrated significantly lower values of both the maximum traction force and the first peak of the traction force and needed significantly longer time to reach the maximum force value in the gl2 test than in the gl1 test. These effects were especially pronounced in cases of the plant surfaces covered with wax projections having higher aspect ratios. The data obtained clearly indicated the impact of waxy plant surfaces on the insect ability to subsequently attach to the clean smooth surface. This effect is caused by the contamination of adhesive pads and experimentally supports the contamination hypothesis.

Mechanoecology: biomechanical aspects of insect-plant interactions.

Plants and herbivorous insects as well as their natural enemies, such as predatory and parasitoid insects, are united by intricate relationships. During the long period of co-evolution with insects, plants developed a wide diversity of features to defence against herbivores and to attract pollinators and herbivores' natural enemies. The chemical basis of insect-plant interactions is established and many examples are studied, where feeding and oviposition site selection of phytophagous insects are dependent on the plant's secondary chemistry. However, often overlooked mechanical interactions between insects and plants can be rather crucial. In the context of mechanoecology, the evolution of plant surfaces and insect adhesive pads is an interesting example of competition between insect attachment systems and plant anti-attachment surfaces. The present review is focused on mechanical insect-plant interactions of some important pest species, such as the polyphagous Southern Green Stinkbug Nezara viridula and two frugivorous pest species, the polyphagous Mediterranean fruit fly Ceratitis capitata and the monophagous olive fruit fly Bactrocera oleae. Their ability to attach to plant surfaces characterised by different features such as waxes and trichomes is discussed. Some attention is paid also to Coccinellidae, whose interaction with plant leaf surfaces is substantial across all developmental stages in both phytophagous and predatory species that feed on herbivorous insects. Finally, the role of different kinds of anti-adhesive nanomaterials is discussed. They can reduce the attachment ability of insect pests to natural and artificial surfaces, potentially representing environmental friendly alternative methods to reduce insect pest impact in agriculture.

The tight attachment achieved by the male discoidal setae is possibly a counter-adaptation to the grease layer on female integument surfaces in green dock beetles.

Green dock beetles Gastrophysa viridula exhibit sexual dimorphism in tarsal attachment setae: females have only pointed, lanceolate and spatula-like setae, while males additionally possess discoidal ones. The sexual dimorphism is probably attributed to the necessity of male discoidal setae to adhere to the smooth back of the female during copulation. We aimed to understand its possible mechanism of attachment with G. viridula. Pull-off forces of both females and males were measured on (i) alive females, (ii) dead and dried females, and (iii) resin replicas of fresh females. The attachment ability tended to increase on dead and replicated female surfaces in both sexes, which indicates that the epicuticular grease layer on the integument of alive intact beetles decreases the attachment. This tendency was prominent in females. The present study clearly showed that in G. viridula discoidal setae enable the males to adhere stronger to female surfaces. The divergent performance found between the sexes differing in their setal composition is probably caused by the stiffness difference between the setae types and by the specific shape of the setal tips. A peculiar reproductive biology in G. viridula is probably attributed to this remarkable divergence of labour in their attachment pads between the sexes.

Petals Reduce Attachment of Insect Pollinators: A Case Study of the Plant Dahlia pinnata and the Fly Eristalis tenax.

In order to understand whether the petal surface in "cafeteria"-type flowers, which offer their nectar and pollen to insect pollinators in an open way, is adapted to a stronger attachment of insect pollinators, we selected the plant Dahlia pinnata and the hovering fly Eristalis tenax, both being generalist species according to their pollinator's spectrum and diet, respectively. We combined cryo scanning electron microscopy examination of leaves, petals, and flower stems with force measurements of fly attachment to surfaces of these plant organs. Our results clearly distinguished two groups among tested surfaces: (1) the smooth leaf and reference smooth glass ensured a rather high attachment force of the fly; (2) the flower stem and petal significantly reduced it. The attachment force reduction on flower stems and petals is caused by different structural effects. In the first case, it is a combination of ridged topography and three-dimensional wax projections, whereas the papillate petal surface is supplemented by cuticular folds. In our opinion, these "cafeteria"-type flowers have the petals, where the colour intensity is enhanced due to papillate epidermal cells covered by cuticular folds at the micro- and nanoscale, and exactly these latter structures mainly contribute to adhesion reduction in generalist insect pollinators.

Coleoptera claws and trichome interlocking.

The present study tests the hypothesis that the specialized claws with a basal tooth found in some coccinellid beetles represent an adaptation to interlock with flexible unbranched trichomes of different plants. We compared the attachment ability of three Coleoptera species, Chnootriba elaterii, Harmonia axyridis (both Coleoptera: Coccinellidae), and Chrysolina herbacea (Coleoptera: Chrysomelidae) with claws of different shape. The attachment ability of insect individuals with or without claws to a plant with leaves bearing straight non-branched trichomes (Cucurbita moschata) and to a plant with smooth leaves (Prunus laurocerasus) was measured in traction force experiments. Insect attachment ability was also tested on a resin replica of C. moschata leaf, to variate trichome stiffness, and on glass as a reference surface. Centrifugal force tester experiments were performed to compare the attachment ability of the two ladybird species to glass and to the leaf of C. moschata. Natural and artificial substrates were characterized in cryo-SEM. The collected data reveal that plant trichomes can enhance insect attachment to plant surface compared with smooth glass by increasing insect friction force, but this is directly related to the trichome stiffness. To effectively grasp soft trichomes, insects evolved special claws-associated structures, such as the dentate claws observed in Coccinellidae.

Effect of Leaf Trichomes in Different Species of Cucurbitaceae on Attachment Ability of the Melon Ladybird Beetle Chnootriba elaterii.

This study investigates the attachment ability of the oligophagous melon ladybird beetle Chnootriba elaterii to leaves of several Cucurbitaceae species. Using cryo-SEM, we described adult and larva tarsal attachment devices and leaf surface structures (glandular and non-glandular trichomes) in Citrullus lanatus, Cucumis melo, Cucumis sativus, Cucurbita moschata, Cucurbita pepo, Ecballium elaterium, Lagenaria siceraria and Luffa aegyptiaca. Using traction force experiments and centrifugal force tests, we measured the friction force exerted by females and larvae on plant leaves. We observed that Cucurbitaceae glandular trichomes do not affect insect attachment ability at both developmental stages, suggesting some adaptation of C. elaterii to its host plants, while non-glandular trichomes, when they are dense, short and flexible, heavily reduce the attachment ability of both insect stages. When trichomes are dense but stiff, only the larval force is reduced, probably because the larva has a single claw, in contrast to the adult having paired bifid dentate claws. The data on the mechanical interaction of C. elaterii at different developmental stages with different Cucurbitaceae species, combined with data on the chemical cues involved in the host plant selection, can help to unravel the complex factors driving the coevolution between an oligophagous insect and its host plant species.

Hierachical epicuticular wax coverage on leaves of Deschampsia antarctica as a possible adaptation to severe environmental conditions.

Using cryo scanning electron microscopy, the surface micromorphology of vegetative (leaf blade and ligule) and generative (pedicel and outer glume) organs in Deschampsia antarctica, one of the only two flowering plants native to Antarctica, was examined. Whereas the pedicel and outer glume were wax-free, both leaf sides had a prominent epicuticular wax coverage consisting of two superimposed layers: polygonal rodlets formed by fused irregular platelets (the lower wax layer) and membraneous platelets (the upper wax layer). Although the adaxial (inner) and abaxial (outer) leaf surfaces showed a similar microstructure of the wax coverage, they differed in the thickness ratio between lower and upper wax layer. The ligule bore a very loose wax coverage composed of separate scale-like projections or clusters of them. We suppose that the two-layered wax densely covering both leaf surfaces might contribute to the plant adaptation to severe environmental conditions in Antarctica due to an increase of its resistance against cold temperatures, icing, harmful UV radiation, and dehydration. The presence of the epicuticular wax on the abaxial leaf side and the ligule as well as the hierarchical structure of the wax coverage on both leaf surfaces is described in D. antarctica for the first time.

Anti-icing strategies of plant surfaces: the ice formation on leaves visualized by Cryo-SEM experiments.

This paper presents an experimental study on surface icing on leaves in six plant species having different surface micromorphology and wettability properties. Contrary to previous studies on ice crystallization, which have been mainly performed by using infrared video thermography, we applied a Cryo-SEM approach allowing not only characterization of plant surfaces in their native conditions but also visualization of ice crystal formation on the native plant surfaces at the micro- and nanoscales. The Cryo-SEM was also used as an experimental device to freeze water vapor, thaw ice crystals, and freeze fluid water on the plant surface again. The experiments clearly demonstrate that trichome coverage (especially with several distinct layers) and 3D wax projections can be recognized as anti-icing strategies of plants. Trichomes can prevent and delay ice formation by being nucleation points for the formation of ice from vapor and protect the plant surface from overcooling, when fluid water freezes in contact with the leaf surface. The study shows for the first time two important effects that might reduce plant cell freezing rate: the presence of air pockets between wax projections that protect from direct contact between ice crystals and the plant cuticle and elimination of fluid water after thawing and preventing further re-freezing on the surface. The detailed knowledge obtained here is not only important for plant ecology, evolution, and plant protection but also for looking for potential biomimetic strategies that reduce/avoid icing of cultural plants and artificial technical surfaces.

Combined Effect of Different Flower Stem Features on the Visiting Frequency of the Generalist Ant Lasius niger: An Experimental Study.

In order to understand the effects of the morphology and surface texture of flower stems in Smyrnium rotundifolium on the visiting frequency of generalist ants, we conducted experiments with Lasius niger ants running on dry wooden sticks mimicking different types of stems: (1) intact (grooved) sticks; (2) sticks painted with slaked (hydrated) lime (calcium carbonate coverage) imitating plant epicuticular wax coverage; (3) intact sticks with smooth polyester plate-shaped cuffs imitating upper leaves; and (4) intact sticks bearing cuffs painted with slaked lime. Ants were attracted by the sweet sugar syrup droplets placed on a stick tip, and the number of ants visiting the drops was counted. Our data showed significant differences in the visiting frequencies between the different types of stem-mimicking samples. The number of recorded ants progressively decreased in the following order of samples: intact sticks-painted sticks-sticks with intact cuffs-sticks with painted cuffs. These results clearly demonstrated that micro/nanoscopic surface coverages and macroscopic physical barriers, especially if combined, have a negative impact on the attractiveness of stems to ants. This study provides further evidence for the hypothesis that having a diversity of plant stems in the field, generalist ants prefer substrates where their locomotion is less hindered by obstacles and/or surface slipperiness.

Surface chemistry of the ladybird beetle adhesive foot fluid across various substrates.

Nature has coevolved highly adaptive and reliable bioadhesives across a multitude of animal species. Much attention has been paid in recent years to selectively mimic these adhesives for the improvement of a variety of technologies. However, very few of the chemical mechanisms that drive these natural adhesives are well understood. Many insects combine hairy feet with a secreted adhesive fluid, allowing for adhesion to considerably rough and slippery surfaces. Insect adhesive fluids have evolved highly specific compositions which are consistent across most surfaces and optimize both foot adhesion and release in natural environments. For example, beetles are thought to have adhesive fluids made up of a complex molecular mixture containing both hydrophobic and hydrophilic parts. We hypothesize that this causes the adhesive interface to be dynamic, with molecules in the fluid selectively organizing and ordering at surfaces with complimentary hydrophobicity to maximize adhesion. In this study, we examine the adhesive fluid of a seven-spotted ladybird beetle with a surface-sensitive analytical technique, sum frequency generation spectroscopy, as the fluid interacts with three substrates of varied wettabilities. The resulting spectra present no evidence of unique molecular environments between hydrophilic and hydrophobic surfaces but exhibit significant differences in the ordering of hydrocarbons. This change in surface interactions across different substrates correlates well with traction forces measured from beetles interacting with substrates of increasing hydrophobicities. We conclude that insect adhesion is dependent upon a dynamic molecular-interfacial response to an environmental surface.

Variation of attachment ability of Nezara viridula (Hemiptera: Pentatomidae) during nymphal development and adult aging.

The present investigation reports data on the attachment ability of the Southern green stink bug Nezara viridula (Hemiptera: Pentatomidae), a relevant pest in the world, along its whole life cycle. Using a centrifugal force tester, we evaluated a) the differences in the attachment ability among the four active nymphal developmental instars (N2-N5 nymphs) and adult to hydrophilic glass, showing an increased attachment ability during ontogenesis, owing to increased pulvilli size and efficiency; b) the possible role of growth and body shape on insect attachment ability on hydrophilic glass during the intermoult period, revealing that N. viridula nymphs attach stronger (higher safety factor) in the first part of the intermoult period; c) the age-specific differences in the attachment ability of adults of both sexes on hydrophilic glass, showing the best performance at an intermediate age, in agreement with a higher proportion of resilin in comparison with younger or older insects; d) the difference in attachment ability on hydrophilic vs. hydrophobic glass along the insect development, revealing a strong effect of surface hydrophobicity on reducing the attachment of N. viridula nymphs and adults. The results on the attachment ability of a hemimetabolous insect along its life cycle are relevant because they 1) shed light on different adaptations of attachment pads in relation to insect size, shape and age; 2) deepen the knowledge on the functional morphological adaptations, thus potentially contributing to the development of suitable control systems for this important pest insect.

Traction force measurements on male Strepsiptera (Insecta) revealed higher forces on smooth compared with hairy substrates.

The aim of this study was to find out how strongly the parasitic insect Stylopsovinae, which has tarsi equipped with tenent hairs and lacking claws, attaches to different substrates. We investigated adhesion of male S. ovinae to the abdomen of its hymenopteran host (Andrena vaga), the hairier abdomen of a Bombus sp. and two artificial smooth reference surfaces with different degrees of hydrophilicity. In our experiments, the male S. ovinae developed significantly higher forces on smooth surfaces. However, the forces were significantly lower on all the hymenopteran surfaces used in the experiment. The absence of anisotropy in the force grip in cranial/caudal direction relative to the host might indirectly indicate that S. ovinae generate forces by adhesion rather than mechanical interlocking with the host hairs. The tolerance of the attachment system of S. ovinae to the substrate chemistry might be explained by the primary contribution of van der Waals interactions and not capillary forces to adhesion in S. ovinae.

Insect-inspired architecture to build sustainable cities.

Materials, structures, surfaces and buildings of insects are of a great scientific interest, but such basic knowledge about the functional principles of these structures is also highly relevant for technical applications, especially in architecture. Some of the greatest challenges for today's architecture are multifunctionality, energy saving and sustainability - problems that insects have partially solved during their evolution. Entomologists have collected a huge amount of information about the structure and function of such living constructions and surfaces. This information can be utilized in order to mimic them for applications in architecture. The main technology areas, in which insect-inspired ideas can be applied, are the following: (1) new materials, (2) constructions, (3) surfaces, (4) adhesives and bonding technology, (5) optics and photonics. A few selected examples are discussed in this short review, but having more than one million described insect species as a source for inspiration, one might expect many more ideas from entomology for insect-inspired biomimetics in architecture. The incorporation of additional knowledge from insect biology into architecture will improve performance of future buildings. However, biologists still do not have a complete understanding of structure-function relationship of insect materials and construction. Hence, many technological areas will benefit from additional basic entomology research. Also the screening for new inspirations from insects is likely to remain an important research field in the near future.

Role of Fruit Epicuticular Waxes in Preventing Bactrocera oleae (Diptera: Tephritidae) Attachment in Different Cultivars of Olea europaea.

The olive fruit fly Bactrocera oleae (Diptera: Tephritidae) is the major pest of cultivated olives (Olea europaea L.), and a serious threat in all of the Mediterranean Region. In the present investigation, we demonstrated with traction force experiments that B. oleae female adhesion is reduced by epicuticular waxes (EWs) fruit surface, and that the olive fruit fly shows a different ability to attach to the ripe olive surface of different cultivars of O. europaea (Arbequina, Carolea, Dolce Agogia, Frantoio, Kalamata, Leccino, Manzanilla, Picholine, Nostrale di Rigali, Pendolino and San Felice) in terms of friction force and adhesion, in relation with different mean values of olive surface wettability. Cryo-scanning morphological investigation revealed that the EW present on the olive surface of the different analyzed cultivars are represented by irregular platelets varying in the orientation, thus contributing to affect the surface microroughness and wettability in the different cultivars, and consequently the olive fruit fly attachment. Further investigations to elucidate the role of EW in olive varietal resistance to the olive fruit fly in relation to the olive developmental stage and environmental conditions could be relevant to develop control methods alternative to the use of harmful pesticides.

Mechanical ecology of fruit-insect interaction in the adult Mediterranean fruit fly Ceratitis capitata (Diptera: Tephritidae).

Fruit features represent a trade-off between dispersal and protection against frugivore insects. To prevent insect attack, plants evolved chemical and physical barriers, mainly studied in leaves, while limited knowledge is available for fruits, especially concerning mechanical barriers. We used the Mediterranean fruit fly to shed light on the mechanical ecology of insect-fruit attachment in a pest species. We tested the following hypotheses: is there any sexual dimorphism in attachment devices and attachment ability? Can the attachment ability of females of Ceratitis capitata to fruits of various host plants vary according to fruit surfaces with different morphology (smooth, hairy, waxy) or physico-chemical properties? The tarsal attachment devices were studied using Cryo-SEM and TEM. The maximum friction forces of C. capitata females on fruit surfaces of typical host plants were evaluated using a load cell force transducer. The attachment ability of both sexes on artificial surfaces was evaluated using a centrifugal force tester. Our data revealed sexual dimorphism in the size of pulvilli, which are wider in females. A higher friction force is exerted by females in comparison with males, in agreement with the need to firmly adhere to the host plant fruit during oviposition. Among the tested fruits, the stronger friction force was recorded on hairy or rough surfaces while a force reduction was recorded on waxy fruits. To unravel the mechanical ecology of insect-plant interaction between plants and species of Tephritidae can be useful to develop non-chemical methods to control these important crop pests.

Attachment ability of females and males of the ladybird beetle Cryptolaemus montrouzieri to different artificial surfaces.

To test the effects of surface chemistry and surface roughness on the attachment ability of female and male Cryptolaemus montrouzieri beetles that are equipped with hairy adhesive pads, traction force experiments were performed on three artificial substrates: (1) hydrophobic smooth glass, (2) hydrophobic smooth epoxy resin, and (3) hydrophobic microrough epoxy resin. Also the micromorphology of the dorsal body side and adhesive pads in males and females was examined using a scanning electron microscopy. The traction force ranged from 0.13 to 3.60 mN in females and from 0.28 to 3.20 mN in males. The force values obtained on different test substrates showed highly significant differences and decreased in the following order of substrates: glass - smooth epoxy resin - microrough epoxy resin. In both females and males, the effect of surfaces was similar. The obtained results clearly showed that both surface parameters, chemistry and roughness, affected the attachment ability of beetles. Similar microstructure of adhesive pads in both sexes resulted in similar attachment performance of males and females on all test substrates.

Egg-laying job makes males hot: body temperature measurements in egg-laying tandems of the dragonfly Sympetrum vulgatum using IR camera.

Tandem oviposition, where males guide females in contact, requires quite a substantial muscle activity from males and, therefore, stronger heat production within the male thorax compared to the female thorax. In the present study, an infrared camera equipped with a macrolens was applied in the field, in order to estimate temperature in different body regions of male and female dragonflies Sympetrum vulgatum laying eggs in tandems. In both sexes, the thorax was considerably warmer than other body parts. The male thorax was on average 3-4 °C warmer than that of the female. These observations support previous data that Sympetrum males have a stronger muscular activity and heat production in tandem during the egg-laying process compared to females. The data provide additional evidence that this kind of contact guarding behavior during oviposition is rather costly for males. The tip of the male abdomen was much warmer than its own abdomen in the middle region. This result might be explained by possible heat transfer from the female head to the male abdomen through the contact between male abdominal appendages and the female head. An alternative explanation might be strong activity of the muscles controlling male anal appendages. Finally, this study also demonstrated a strong potential of IR cameras in field studies of dragonfly behavioral physiology.

Porous substrate affects a subsequent attachment ability of the beetle Harmonia axyridis (Coleoptera, Coccinellidae).

According to literature data, porous substrates can cause a reduction of insect attachment ability. We carried out traction experiments with adult ladybird beetles Harmonia axyridis on the smooth solid glass sample and rough porous Al2O3 membrane to prove the primary effect of absorption of the insect pad secretion by porous media, rather than surface roughness, on the attachment force on the porous sample. With each insect individual, a set of five experiments was conducted: (1) on glass; (2) on the porous membrane; (3-5) on glass immediately after the test on the porous surface, then after 30 min and 1 h of recovery time. On the porous substrate, the forces, being similar in females and males, were greatly reduced compared to those measured on glass. A significant difference between the force values obtained in the first (before the test on the porous sample) and second (immediately after the experiment on the porous sample) tests on glass was observed. After 30 min recovery time, beetles completely regained their attachment ability. Females produced significantly lower forces than males in all experiments on glass: the differences are probably caused by the sexual dimorphism in the microstructure of their adhesive pads. The obtained results are of fundamental importance for further application in biomimetics of novel insect-repelling surfaces and in plant protection by using porous materials.

Investigating the efficiency of a bio-inspired insect repellent surface structure.

Most insects with smooth or hairy adhesive pads have very little problems in attaching to smooth substrates. A careful selection of surface roughness, however, can effectively limit the contact area of the adhesive organs with the surface. In comparison to conventional toxin-based insect repelling methods, biologically inspired micro- and nanostructured insect repellent surface structures, thus, offer a great potential to effective and environmentally-friendly control insect pests. We here present a simple experimental approach to qualitatively and quantitatively analyse the efficiency of a micro- and nanorough surface structure. Nauphoeta cinerea and Gastrophysa viridula as model organisms for insects with smooth and hairy adhesive pads, respectively, were placed in an enclosed environment. The escape movements of freely running insects over either the repellent or a control surfaces were counted and analyzed in detail. The tested surface structures were able to significantly reduce the escape of cockroaches with smooth adhesive pads by 44.1%. Interestingly, the data indicates that N. cinerea might use mechanical cues from the antenna to discriminate between surfaces before making contact with the adhesive pads. G. viridula with hairy adhesive pads were not significantly affected by the surface structure. By carefully adjusting 'critical' surface topography parameters relevant for insect adhesion, more efficient and selective repellent surfaces might be achieved. Such nanostructure-based insect repellent surfaces could also help to utilize recruitment behavior of certain insect species and might present a novel approach to effectively control insect pests.