Senta Niederegger

Structural Design and Biomechanics of Friction-Based Releasable Attachment Devices in Insects

Abstract Design of attachment devices in insects varies enormously in relation to different functional loads. Many systems, located on different parts of the body, involve surfaces with particular frictional properties. Such systems evolved to attach parts of the body to each other, or to attach an insect to the substratum by providing fast and reversible attachment/detachment. Among these systems, there are some that deal with predefined surfaces, and others, in which one surface remains unpredictable. The first type of system occurs, for example, in wing-locking devices and head-arresting systems and is called probabilistic fasteners. The second type is mainly represented by insect attachment pads of two alternative designs: hairy and smooth. The relationship between surface patterns and/or mechanical properties of materials of contact pairs results in two main working principles of the frictional devices: mechanical interlocking, or maximization of the contact area. We give an overview of the functional design of two main groups of friction-based attachment devices in insects: probabilistic fasteners and attachment pads.

Friction and adhesion in the tarsal and metatarsal scopulae of spiders.

Friction and adhesion forces of the ventral surface of tarsi and metatarsi were measured in the bird spider Aphonopelma seemanni (Theraphosidae) and the hunting spider Cupiennius salei (Ctenidae). Adhesion measurements revealed no detectable attractive forces when the ventral surfaces of the leg segments were loaded and unloaded against the flat smooth glass surface. Strong friction anisotropy was observed: friction was considerably higher during sliding in the distal direction. Such anisotropy is explained by an anisotropic arrangement of microtrichia on setae: only the setal surface facing in the distal direction of the leg is covered by the microtrichia with spatula-like tips. When the leg is pushed, the spatula-shaped tips of microtrichia contact the substrate, whereas, when the leg is pulled over a surface, setae bend in the opposite direction and contact the substrate with their spatulae-lacking sides. In an additional series of experiments, it was shown that desiccation has an effect on the friction force. Presumably, drying of the legs results in reduction of the flexibility of the setae, microtrichia, spatulae, and underlying cuticle; this diminishes the ability to establish proper contact with the substrate and thus reduces the contact forces.

Beetles and flies collected on pig carrion in an experimental setting in Thuringia and their forensic implications.

Decomposition processes and insect succession were analysed on a total of eight pig (Sus scrofa) carcasses. The survey was carried out in four different periods between November 2007 and August 2008 and on two different substrates (meadow, concrete floor close to a building). The experiments were placed in a rural site in the surroundings of Jena, Thuringia. The duration and specificity of the different decomposition stages were dependent on season, weather and quality and quantity of insect colonization. Whereas the carrion maintained a bloated appearance even after 133 days in winter, it reached the dry stage within 8 days in the summer months. The type of substrate had few effects on the decaying process, but the insects were generally more abundant on the meadow. In total, more than 57 species of Diptera belonging to 17 families and 48 species of Coleoptera belonging to 14 families were identified. Dominant species belonged to the families of Calliphoridae (n = 11 spp.), Sarcophagidae (n = 8), Muscidae (n = 9), Piophilidae (n = 3), Silphidae (n = 6), Dermestidae (n = 3), Nitidulidae (n = 4), Cleridae (n = 3) and Histeridae (n = 2). Remarkably, the rather common Nicrophorus species were completely absent, whereas the usually rare Necrodes littoralis (L.) was present in larger numbers. No distinct coincidence between the occurrence of a single species and a certain decomposition stage could be confirmed. A main objective of the study is the establishment of a forensic entomological database for Central Europe, especially Thuringia.

From behavior to fictive feeding: anatomy, innervation and activation pattern of pharyngeal muscles of Calliphora vicina 3rd instar larvae.

A description of the muscles and nerves involved in feeding of larval Calliphora vicina is given as a prerequisite to establish fictive feeding patterns recorded from the isolated central nervous system. Feeding Diptera larvae show a repetitive sequence of pro- and retraction of the cephalopharyngeal skeleton (CPS), elevation and depression of the mouth hooks and food ingestion. The corresponding pharyngeal muscles are protractors, mouth hook elevators and depressors, the labial retractor and cibarial dilator muscles. These muscles are innervated by the prothoracic accessory nerve (PaN), maxillary nerve (MN) and antennal nerve (AN) as shown electrophysiologically by recording action potentials from the respective nerve that correlate to post-synaptic potentials on the muscles. All three nerves show considerably more complex branching patterns than indicated in the literature. Extracellular recordings from the stumps of PaN, MN and AN connected to an isolated CNS show spontaneous rhythmic motor patterns that reflect the feeding sequence in intact larvae. Variability of the feeding pattern observed in behavioral experiments is also evident from the level of motor output from an isolated CNS. The data obtained from Calliphora will facilitate electrophysiological investigations dealing with the genetic background of feeding behavior in Drosophila larvae.

The brain can eat: establishing the existence of a central pattern generator for feeding in third instar larvae of Drosophila virilis and Drosophila melanogaster.

To establish the existence of a central pattern generator for feeding in the larval central nervous system of two Drosophila species, the gross anatomy of feeding related muscles and their innervation is described, the motor units of the muscles identified and rhythmic motor output recorded from the isolated CNS. The cibarial dilator muscles that mediate food ingestion are innervated by the frontal nerve. Their motor pathway projects from the brain through the antennal nerves, the frontal connectives and the frontal nerve junction. The mouth hook elevator and depressor system is innervated by side branches of the maxillary nerve. The motor units of the two muscle groups differ in amplitude: the elevator is always activated by a small unit, the depressor by a large one. The dorsal protractors span the cephalopharyngeal skeleton and the body wall hence mediating an extension of the CPS. These muscles are innervated by the prothoracic accessory nerve. Rhythmic motor output produced by the isolated central nervous system can simultaneously be recorded from all three nerves. The temporal pattern of the identified motor units resembles the sequence of muscle contractions deduced from natural feeding behavior and is therefore considered as fictive feeding. Phase diagrams show an almost identical fictive feeding pattern is in both species.

Influence of food substrates on the development of the blowflies Calliphora vicina and Calliphora vomitoria (Diptera, Calliphoridae)

The blowflies Calliphora vicina and Calliphora vomitoria are among the first colonizers of human remains in Europe. Laboratory development studies with immature stages of these blowflies for postmortem interval (PMI) calculations are generally performed on different media such as processed food substrates or liver of various animals. The question arises whether these media per se influence the development of larvae and thus PMI calculations? In this systematic approach, the effects of an assortment of food substrates on the development of the larvae were analyzed. C. vomitoria showed much better growth on processed substrates such as beef, pork, turkey, and mixed minced meats than on unprocessed substrates such as beef and pork liver and turkey steak. Beef liver even impeded full development of the species and resulted in death of all individuals. C. vomitoria was therefore categorized as a specialist. Even though mixed minced meat yielded low pupariation rates for C. vicina, the species showed, otherwise, comparable growth rates on all substrates tested and was thus considered to be a generalist. These findings emphasize the importance of parameters besides temperature on the development rates of forensically important fly larvae.

Cuticular muscle attachment sites as a tool for species determination in blowfly larvae

First results of a new method for species determination in third instar larvae of saprophagous blowflies are introduced. Cuticular attachment sites of a limited number of transversal muscles are visualized for light microscopic analysis. After removing the muscles and staining the cuticle, the attachment sites become visible as laterally symmetrical segmental clusters of dark dots. The combined patterns of five such clusters, located in the second, third and fourth segments, show sufficient differences to allow reliable separation of externally very similar larval Lucilia sericata and Lucilia illustris as well as Calliphora vomitoria and Calliphora vicina, the most common saprophagous blowfly species in Europe. Species determination even in poorly conserved, discoloured and fragmented blowfly larvae becomes possible with this new method. The method can primarily be applied for postmortem interval (PMI) calculations in forensic entomology. Interspecific morphological similarity of the larvae and differences in growth rate make species determination an essential requisite for an exact PMI calculation.

Feel the heat: The effect of temperature on development, behavior and central pattern generation in 3rd instar Calliphora vicina larvae

Like in all poikilothermic animals, higher temperatures increase developmental rate and activity in Calliphora vicina larvae. We therefore could expect temperature to have a persistent effect on the output of the feeding and crawling central pattern generators (CPGs). When confronted with a steep temperature gradient, larvae show evasive behavior after touching the substrate with the cephalic sense organs. Beside this reflex behavior the terminal- and dorsal organ might also mediate long term CPG modulation. Both organs were thermally stimulated while their response was recorded from the maxillary- or antennal nerve. The terminal organ showed a tonic response characteristic while the dorsal organ was not sensitive to temperature. Thermal stimulation of the terminal organ did not affect the ongoing patterns of fictive feeding or crawling, recorded from the antennal- or abdominal nerve respectively. A selective increase of the central nervous system (CNS) temperature accelerated the motor patterns of both feeding and crawling. We propose that temperature affects centrally generated behavior via two pathways: short term changes like thermotaxis are mediated by the terminal organ, while long term adaptations like increased feeding rate are caused by temperature sensitive neurons in the CNS which were recently shown to exist in Drosophila larvae.

See the light: electrophysiological characterization of the Bolwig organ's light response of Calliphora vicina 3rd instar larvae.

The anatomy and development of the larval cyclorraphous Diptera visual system is well established. It consists of the internal Bolwig organ (BO), and the associated nerve connecting it to the brain. The BO contributes to various larval behaviors but was never electrophysiologically characterized. We recorded extracellulary from the Bolwig nerve of 3rd instar Calliphora vicina larvae to quantify the sensory response caused by BO stimulation with light stimuli of different wavelengths, intensities and directions. Consistent with previous behavioral experiments we found the BO most sensitive to white and green, followed by blue, yellow, violet and red light. The BO showed a phasic-tonic response curve. Increasing light intensity produced a sigmoid response curve with an approximate threshold of 0.0105 nW/cm(2) and a dynamic range from 0.105 nW/cm(2) to 52.5 nW/cm(2). No differences exist between feeding and wandering larvae which display opposed phototaxis. This excludes reduced BO sensitivity from causing the switch in behavior. Correlating to the morphology of the BO frontal light evoked the maximal reaction, while lateral light reduced the neural response asymmetrically: Light applied ipsilaterally to the recorded BO always produced a stronger response than when applied from the contralateral side. This implies that phototacic behavior is based on a tropotactic mechanism.

The thoracic muscular system and its innervation in third instar Calliphora vicina Larvae. I. Muscles of the pro- and mesothorax and the pharyngeal complex

An anatomical description is given by the muscles in the pro‐ and mesothorax, and those associated with the feeding apparatus (cephalopharyngeal skeleton, CPS) that participate in feeding behavior in third instar Calliphora larvae. The body wall muscles in the pro‐ and mesothoracic segments are organized in three layers: internal, intermedial, and external. The muscles were labeled with roman numerals according to the nomenclature in use for the abdominal segments. Muscles associated with the CPS are labeled according to their function. The prothorax bears five pairs of lateral symmetrically longitudinal segmental body wall muscles and lacks the transversal muscle group present in the mesothorax and abdominal segments. Additionally, four pairs of intersegmental muscles project from the prothorax to the second, fourth, and fifth segment. The mesothorax bears 15 pairs of segmental longitudinal and 18 pairs of transversal muscles. The accessory pharyngeal muscles span the CPS and the cuticle. Three pairs of protractors and retractors and two pairs of mouth hook accessors (MHAC) exist, which move the CPS relative to the body. The pharyngeal muscles are exclusively attached to the structures of the CPS. The mouth hook elevators and depressors, which mediate the hooks rotation are attached to the ventral arm of the CPS and project to a dorsal (elevators) or ventral (depressors) protuberance of the mouth hooks. The cibarial dilator muscles (CDM) span the dorsal arms of the CPS and the dorsal surface of the esophagus and mediate food ingestion. The labial retractors (LRs) lack antagonists and project from the ventral surface of the CPS to the unpaired labium. Contractions of these muscles open the mouth cavity. J. Morphol. 271:960–968, 2010.