Hemolymph Pressure Research Articles

Proline metabolism and its osmotic regulatory role in enhancing hypersalinity tolerance of the razor clam (Sinonovacula constricta)

Proline serves as an important osmolyte in maintaining osmotic pressure and stabilizing cellular structure in aquatic organisms, and is essential for adaptation to salinity stress. This study focuses on the razor clam (Sinonovacula constricta), and investigated the metabolism and synthesis of proline in response to changing salinity. The proline content in gill tissues of razor clams significantly increased over time and with different levels of salinity exposure. The key genes involved in proline metabolism, including (Aminotransferase (ScOAT), Δ1-pyrroline-5-carboxylic acid synthase (ScP5CS), and proline dehydrogenase (ScProDH) were investigated, as well as enzyme activities associated with these genes in razor clams. The expression and activity of P5CS increased significantly under hypersalinity conditions, whereas that of OAT enzyme activity decreased. This suggested that the glutamate pathway had an important role in the proline synthesis during hypersalinity. ProDH activity was negatively correlated with salinity and regulated by the proline content, indicating a significant role of ProDH in proline accumulation under hypersalinity conditions. ScP5CS was heterologously expressed in yeast Saccharomyces cerevisiae, followed by exogenous proline supplementation studies. Recombinant S. cerevisiae incubated with ScP5CS exhibited elevated levels of proline, with the growth performance of recombinant strain surpassing that of the control group after salinity stress. In addition, razor clams exposed to exogenous proline under high salinity conditions showed expedite osmotic regulation, whereby the hemolymph pressure reached equilibrium more quickly. Together, these results indicated that the proline content increased under high salinity, which could improve the tolerance of razor clams to hypersalinity, offering novel perspectives into molecular mechanism of salinity adaptation in shellfish.

Metabolites, ions, and the mechanisms behind seasonal cold hardening of Pyrochroa coccinea (Pyrochroidae) larvae

The larvae of the black headed cardinal beetle Pyrochroa coccinea, overwinters under the bark of dead logs in northern European dioecious forests, and are thus exposed to temperatures below the melting point of their bodily fluids. Here we explore the mechanisms behind their seasonal cold hardening by characterising field samples collected monthly throughout the year. Both the lower lethal temperature and supercooling point dropped as much as 10℃ in the second half of November, reaching values around –15℃ by the beginning of December. This change was accompanied by a 320 mosmol/kg increase in hemolymph osmolality, which is a doubling compared to the summer levels. We used NMR metabolomics to identify and measure the absolute concentrations of the responsible cryoprotective C-H containing metabolites in the hemolymph. The largest increase was found to be in either glucose or trehalose, with an average total increase of 120 mM. Proline, alanine, and choline concentrations were found to increase by around 10 mM each. Contrarily, phosphocholine and phosphoethanolamine were halved, resulting in a total decrease of around 50 mM. These measurements were complemented with ion exchange chromatography measurements. This allowed us to account for all the osmotic pressure in the summer hemolymph, and the measured concentration changes explained as much as 40 % of the observed osmolality increase upon cold hardening. Preliminary results indicate that the remainder may be explained by non-colligative protein contributions.

First 3-D reconstruction of copulation in Lepidoptera: interaction of genitalia in Tortrix viridana (Tortricidae)

BackgroundThe process of copulation in Lepidoptera is understudied and poorly understood from a functional perspective. The purpose of the present paper is to study the interaction of the male and female genitalia of Tortrix viridana Linnaeus, 1758 via three-dimensional models of pairs fixed during copulation. Other techniques (confocal laser scanning microscopy, scanning electron microscopy and histology) were used to clarify the role of the organs involved in the process.ResultsThree-dimensional models based on micro-CT scanned copulating pairs were generated allowing visualisation of the position of the male and female counterparts, spatial changes during copulation, and the skeleto-muscular apparatus involved in the process. The male genitalia and their musculature are simplified in comparison with other lineages of the family, but the opposite is true for the female genitalia. The attachment of the couple is achieved only through flexion of the valvae, clasping the large and sclerotised sternite 7 of the female. The anal cone and socii of the male are in contact with certain parts of the anal papillae and the sterigma of the female. The long tubular vesica is inserted into the narrow posterior part of the ductus bursae. Its eversion is achieved by an increase in haemolymph pressure. A possible mechanism of stimulation of the female via pulsations of the diverticulum of the vesica was discovered. A compressed sclerotised area of the ductus bursae putatively serves as a valve controlling the transfer of ejaculated materials. Copulation progresses through two phases: in the first the vesica and its diverticulum are inflated by haemolymph, and in the second the diverticulum is not inflated, and the vesica is occupied by viscous ejaculated material. The formation of the multilayered spermatophore was observed, and we discovered that sperm is transferred very late in the copulation process.ConclusionsCopulation process in Lepidoptera is studied for the first time with three-dimensional reconstructions of couples of Tortrix viridana, used as a model species. The internal genitalia is the scenario of multiple interactions between male and female, but the external remain static. A possible mechanism of stimulation of the female internal copulation organs is proposed.

On latches in biological systems: a comparative morphological and functional study of the retinaculum and the dens lock in Collembola

BackgroundSpringtails have the ability to jump using morphological structures consisting of a catapult, the furca, and a latching system constructed with interaction of the retinaculum and the dens lock. The retinaculum engages in the furca at the dens lock in order to form a spring mechanism. They exhibit diversified morphological traits that serve as adaptations to a variety of terrestrial strata and aquatic surface environments. This comparative morphofunctional study centered on the retinaculum and the furcular region of the dens lock aims to describe the morphological variation between taxa and provide insights into the functional dynamics of the latching mechanism at work in the jumping apparatus. Using SEM, µCT and cLSM, we compared representatives of Collembola taxa, Poduromorpha (Neanura muscorum and Podura aquatica), Symphypleona (Dicyrtomina ornata) and Neelipleona (Megalothorax minimus), and examined extracts of the environment in which they were collected.ResultsA retinaculum is absent in N. muscorum, although vestigial muscles were found. Abdominal musculature varies significantly, being more abundant in springtails with clear segmentation (N. muscorum and P. aquatica), and reduced in springtails with fused segmentation (D. ornata and M. minimus). The M.a-ret varies as regards architecture and point of connection with the ramus, which is lateral in P. aquatica and median in the other species studied. The number of teeth in the retinaculum ramus also varies between three in M. minimus and four in the other species. The dens lock of all species studied has two locks and two furrows.ConclusionsThe retinaculum and dens lock interact in a key-lock relationship. The latching and unlatching mechanism from the retinaculum and dens lock appear to be similar in all the taxa examined, occurring by muscle force. This leads us to question the hypothesis that hemolymph pressure may be a force generator in jumping. We offer a reconstruction of the ground pattern of the retinaculum and dens lock and, in addition, an explanation of their functioning and the interaction between them. Finally, we frame the interaction between the retinaculum and the dens lock as a latch in a biological system, a mechanism which functions by force of physical contact.

Neuroanatomy and functional morphology of peripheral receptor neurones with direct projections into the protocerebrum of the brains of the locust and a jewel beetle

AbstractProjections from peripheral receptors directly into the protocerebrum of insects have only been little studied. Retrograde staining of nerves from the antennae, maxillary palps and legs has revealed some fibres that project into the central areas of the protocerebrum. In the case of the antennae and palps, it was not known which receptors were responsible for these projections. In the legs of locusts, multipolar neurons (MN) with characteristic terminal dendritic masses (TDM) have been described to project into a neuropil called “superior ventral inferior protocerebrum” (SVIP). However, such neurons have only been found in the abdominal infrared organs of the Australian fire beetle Merimna atrata, where they function as thermoreceptors. In several orthopterans, fibres from the antennae and palps also project into the SVIP. The present work suggests that the multipolar neuron from the infrared organ of Merimna also projects into the protocerebrum, possibly into a ventral region functionally analogous to the SVIP. No MNs but single scolopidia were found in the tips of the antennae and palps of locusts, apparently responsible for projections into the SVIP, where they probably function as receptors for haemolymph pressure.

Spider‐Inspired, Fully 3D‐Printed Micro‐Hydraulics for Tiny, Soft Robotics

AbstractMany arachnids use internal hemolymph pressure to actuate extension in their leg joints. The inherent large foot displacement‐to‐body length ratio that arachnids can achieve through hydraulics relative to muscle‐based actuators is both energy and volumetrically efficient. Until recent advances in nano/microscale 3D printing with two‐photon polymerization (2PP), the physical realization of synthetic complex ‘soft’ joints would have been impossible to replicate and fill with a hydraulic fluid into a sealed sub‐millimeter system. Inspired by nature, the smallest scale 3D‐printed hydraulic actuator (4.9 × 10−4 mm3) by more than an order of magnitude is demonstrated. The use of stiff 2PP polymers with micron‐scale dimensions enable compliant membranes similar to exoskeletons seen in nature without the requirement for low‐modulus materials. The bio‐inspired system is designed to mimic similar hydraulic pressure‐activated mechanisms in arachnid joints utilized for large displacement motions relative to body length. Using variations on this actuator design, the ability to transmit forces with relatively large magnitudes (milliNewtons) in 3D space is demonstrated, as well as the ability to direct motion that is useful towards microrobotics and medical applications. Microscale hydraulic actuation provides a promising approach toward the transmission of large forces and 3D motions at small scales, previously unattainable in wafer‐level 2D microelecromechanical systems (MEMS).

From the Ethiopian Bale Mountains hotspotFilopalpinae subfam. nov., a new taxon of Laniatorean harvestmen based on external and genital morphology (Arachnida, Opiliones, Assamiidae).

Filopalpinae subfam. nov. is proposed as a new subfamily within family Assamiidae based on a new genus, Filopalpus gen. nov. and on five new species: F. joschmidti sp. nov., F. bale sp. nov., F. kakaensis sp. nov., F. altomontanus sp. nov. and F. niger sp. nov. All originate from the highlands of Ethiopia, namely the Bale Mountains and adjacent volcanoes in Oromia State. Species of the new subfamily are characterized by dense body cover with pointed tubercles causing a hedgehog appearance of the body (dorsal side, partly ventral side), extreme sexual dimorphism in pedipalps (in males thread-like long and thin surpassing body length by about four to six times, devoid of apophyses and thorns; in females short and stout about body length, with a distal grasping hand formed by apophyses and strong setae of tibia and tarsus, and tarsal claw). Male genital morphology is characterized by a hemolymph-pressure system of truncus penis and glans, a dorsal sub-distal tube-like glans with an inner prickly funnel, which is everted by hemolymph pressure during courtship. These male genital characters place the present species in Assamiidae, but external morphology points to a strongly different separate evolutionary unit within this family. The five species known to date were secured at altitudes between 2830 and 4100 m. At least two of the collecting areas are now devoid of forest due to human impact. To secure the present specimens at high-altitude localities was possible only by searching in remote microhabitats unavailable for cattle and man-made fire. A perspective on current subfamilies of Assamiidae and on pedipalpal morphology and evolution is presented.

Physiological and Population Responses of Nilaparvata lugens after Feeding on Drought-Stressed Rice.

Simple SummaryDrought is considered a critical threat to crop growth and sustainable agriculture worldwide, and it also greatly impacts insect development and population growth. Brown planthopper (BPH), Nilaparvata lugens (Stål), is the predominant rice crop pest in China, and the damaging effects of BPH are enhanced by its strong migratory and reproductive capacities. Our results provide useful information about the effect of drought stress on the poor population growth and negative physiological changes in BPH. Negative changes to water balance and osmotic pressure can cause a decline in the quality of BPH; the GST content of BPH feeding on drought-stressed rice was significantly higher than BPH feeding on non-stressed control plants, and the length of flight muscle sarcomeres and mitochondrial content were decreased in BPH feeding on drought-stressed rice. These findings suggest that water management greatly impacts the physiology and population growth of BPH, and provide a basis for understanding physiological and population-wide responses in BPH during drought stress, which may be helpful in understanding the relationship between drought stress and BPH infestation. Drought stress greatly impacts insect development and population growth. Some studies have demonstrated increased reproductive capacity in drought-stressed insects; however, physiological changes in the brown planthopper (BPH), Nilaparvata lugens (Stål), during periods of drought are unclear. In this study, BPH fed on drought- stressed rice had lower population numbers than BPH feeding on non-stressed rice. Water content, osmotic pressure of hemolymph and total amino acid content of BPH were significantly lower when BPH fed on drought-stressed rice compared to the non-stressed control; however, glucose content and glutathione S-transferase (GST) activity were significantly higher in BPH fed on drought-stressed rice. The expression of Vitellogenin and Exuperantia in BPH fed on drought-stressed rice was higher than that in BPH feeding on non-stressed control plants. The size of myofibrils and the abundance of mitochondria in BPH flight muscles were significantly lower in BPH fed on drought-stressed rice compared to non-stressed plants. These results indicate that water management impacts the physiology of BPH, which may be useful in understanding the relationship between drought stress and this damaging herbivore.

Slit sense organ distribution on the legs of two species of orb-weaving spider (Araneae: Araneidae)

Biotic and abiotic mechanical stimuli are ubiquitous in the environment, and are a widely used source of sensory information in arthropods. Spiders sense mechanical stimuli using hundreds of slit sense organs (small isolated slits, large isolated slits, groups of slits and lyriform organs) distributed across their bodies and appendages. These slit sense organs are embedded in the exoskeleton and detect cuticular strain. Therefore, the spatial pattern of these sensors can give clues into how mechanical stimuli from different sources might be processed and filtered as they are transmitted through the body. Here, we map the distribution of slit sense organs on the legs in two species of orb-weaving spider, A. diadematus and T. edulis, in which slit sense organ distribution has not previously been investigated. We image the spiders’ legs using scanning electron microscopy, and trace the position and orientation of slits on these images to describe the distribution and external morphology of the slit sense organs. We show that both species have a similar distribution of slit sense organs, with small isolated slits occurring in consistent lines parallel to the long axis of the legs, whilst large isolated slits, groups of slits and lyriform organs appear in fixed positions near the leg joints. Our findings support what has been described in the literature for several other species of spider, which indicates that slit organ arrangement is conserved across spiders in different evolutionary lineages and with disparate hunting strategies. The dispersed distribution of small isolated slits along the whole length of the leg may be used to detect large-scale strain of the leg segment as a result of muscle activity or internal changes in haemolymph pressure.

MaNmrA, a Negative Transcription Regulator in Nitrogen Catabolite Repression Pathway, Contributes to Nutrient Utilization, Stress Resistance, and Virulence in Entomopathogenic Fungus Metarhizium acridum.

Simple SummaryNutrient metabolism is closely related to the growth, development, and pathogenicity of pathogenic fungi. The nitrogen catabolite repression (NCR) pathway is a well-known fungal nitrogen source regulation path, in which NmrA plays an important regulatory role. Here, we reported a negative regulatory protein MaNmrA, the NmrA homologous protein, in the entomopathogenic fungus Metarhizium acridum, and found that it played important roles in carbon and nitrogen metabolism, growth, stress tolerance, and virulence of M. acridum. Our work will provide a theoretical basis for further exploring the functions of NCR pathway related genes in entomopathogenic fungi.The NCR pathway plays an important regulatory role in the nitrogen metabolism of filamentous fungi. NmrA, a central negative regulatory protein in the NCR pathway and a key factor in sensing to the carbon metabolism, plays important roles in pathogenic fungal nutrition metabolism. In this study, we characterized the functions of MaNmrA in the insect pathogenic fungus M. acridum. Multiple sequence alignments found that the conserved domain (NAD/NADP binding domain) of MaNmrA was highly conservative with its homologues proteins. Deletion of MaNmrA improved the utilization of multiple carbon sources (such as glucose, mannose, sucrose, and trehalose) and non-preferred nitrogen sources (such as NaNO3 and urea), significantly delayed the conidial germination rate and reduced the conidial yield. The MaNmrA-disruption strain (ΔMaNmrA) significantly decreased tolerances to UV-B and heat-shock, and it also increased the sensitivity to the hypertonic substance sorbitol, oxygen stress substance H2O2, and cell wall destroyer calcofluor white, indicating that loss of MaNmrA affected cell wall integrity, tolerances to hypertonic and oxidative stress. Bioassays demonstrated that disruption of MaNmrA decreased the virulence in both topical inoculation and intrahemocoel injection tests. Further studies revealed that the appressorium formation, turgor pressure, and colonization in hemolymph were significantly reduced in the absence of MaNmrA. Our work will deepen the functional cognition of MaNmrA and make a contribution to the study of its homologous proteins.

A Comparative Study on Low and High Salinity Tolerance of Two Strains of Pinctada fucata

This study compares salinity tolerance between red and black shell Pinctada fucata salinity stress of 20 and 50‰, while 35‰ was used as a control. The hemolymph osmotic pressure, inorganic ion concentration, the activities of Na+-K+ -ATPase, respiratory metabolism related enzymes and liver tissue antioxidant related enzymes were measured at 12 and 24 h after salinity stress. The osmotic pressure and inorganic ion concentration of hemolymph of two strains P. fucata increased significantly with the increase of salinity. The activity of Na+-K+ -ATPase of red P. fucata only decreased under low salinity at 24 h, and was significantly higher than that the control under low salinity at 12 h and high salinity at 12 and 24 h. The succinate dehydrogenase (SDH) activities of the P. fucata treatment groups were significantly higher than those the control at 12 h. The lactate dehydrogenase (LDH) activity increased significantly with salinity at 12 h. and the black P. fucata LDH activity was significantly higher than the control at 24 h, while the LDH activity of red P. fucata was significantly lower than that the control in 50‰ salinity. The superoxide dismutase (SOD) activity of black P. fucata was significantly lower than that the control, while that of red P. fucata was significantly higher than that the control within 24. At 12 h, the catalase (CAT) activity of red P. fucata increased significantly with salinity, but decreased significantly with salinity at 24 h. The CAT activity of black P. fucata was highest at 24 h under low salinity. Glutathione peroxidase (GSH-Px) and alkaline phosphatase (AKP) activities of red P. fucata were significantly higher than those the control under low or high salinity. At high salinity for 24 h, the GSH-Px activity was lowest in black P. fucata, AKP activity was highest. The present study indicates that the physical responses of P. fucata to the salinity stress vary with shell colors. The red P. fucata can quickly respond positively to the change of environmental salinity and reduce the damage caused by the change of environmental salinity.

Physiological and gene expression profiles of leg muscle provide insights into molting-dependent growth of Chinese mitten crab (Eriocheir sinensis)

Molting is a common and periodic process in crustaceans, which make crustacean withdraw from intricate and old exoskeleton, and get an increased body weight after molting. However, the underlying molecular mechanism of internal impetus and physiological features of molting process which impact muscle growth and development is unclear. In this study, the osmotic pressure of hemolymph, histological section, moisture of walking leg muscle, and comparative transcriptome were conducted in Eriocheir sinensis among different molting stages. Osmotic pressure and moisture analysis revealed that a hypertonic environment arose at pre-molt stage, which may induce the initiation of water uptake. The explosive water uptake mainly took place at post-molt stage leading to dramatic weight gain. Comparative transcriptome analysis revealed a total of 2264 differentially expressed genes between pre-molt and post-molt, including 1387 up-regulated genes at pre-molt stage, which negatively regulate muscle growth and development (eg. POMT, NEB2, ZCI2A, and MSTN), and 877 up-regulated genes at post-molt stage which are associated with positive regulation in muscle growth and proliferation (eg. TITIN, mTOR, XRCC, and ICCR). GO enrichment analysis further illustrated that many genes act on proteolysis, ion transport, and transmembrane transport which help with the maintenance of a hypertonic environment in the hemolymph at pre-molt stage. Meanwhile, differentially expressed genes at post-molt stage were enriched in DNA repair and DNA replication indicating the activated muscle growth and development process. Our results provided novel insights into the genetic interplay of osmoregulation, water uptake, and muscle development in E. sinensis during molting.

A mysterious dwarf: Suthepiidae nov. fam., a new harvestman family from mountains of northern Thailand (Arachnida: Opiliones: Laniatores)

A new family of laniatorean harvestmen from northern Thailand is proposed, Suthepiidae fam. nov., which comprises one new genus and one new species, Suthepia inermis sp. nov. This family stands out by characters hitherto unknown or rarely recorded for Opiliones, and close relatives of this taxon are presently not discernible. Important characters are a short and compact penis with a massively enlarged distal part with a rich armament of sclerites and membranes which can be moved and everted by hemolymph pressure during mating; the pedipalp of males and females is without raptorial adaptations, i.e. elevated sockets (= apophyses) carrying strong distal spines are completely absent, therefore no prey capture basket is present; the male pedipalpal tarsus is enlarged and compressed, its dorso-distal double apophysis presumably contains a gland; the pedipalpal claw is weak, not longer or stronger than long hyaline setae on the dorso-distal apophysis of the tarsus. The only specimens presently known were found in litter of a few forested mountain stocks, between 450 m and 1400 m altitude. A brief overview of Southeast Asian laniatorean families is presented.

The control of nocifensive movements in the caterpillar Manduca sexta.

In response to a noxious stimulus on the abdomen, caterpillars lunge their head towards the site of stimulation. This nocifensive 'strike' behavior is fast (∼0.5 s duration), targeted and usually unilateral. It is not clear how the fast strike movement is generated and controlled, because caterpillar muscle develops peak force relatively slowly (∼1 s) and the baseline hemolymph pressure is low (<2 kPa). Here, we show that strike movements are largely driven by ipsilateral muscle activation that propagates from anterior to posterior segments. There is no sustained pre-strike muscle activation that would be expected for movements powered by the rapid release of stored elastic energy. Although muscle activation on the ipsilateral side is correlated with segment shortening, activity on the contralateral side consists of two phases of muscle stimulation and a marked decline between them. This decrease in motor activity precedes rapid expansion of the segment on the contralateral side, presumably allowing the body wall to stretch more easily. The subsequent increase in contralateral motor activation may slow or stabilize movements as the head reaches its target. Strike behavior is therefore a controlled fast movement involving the coordination of muscle activity on each side and along the length of the body.

Diverticula in Male Lycorea halia Butterflies (Lepidoptera: Nymphalidae: Danaini: Itunina)\u2014Support Organs for Everted Hairpencils with Unique Ultrastructure

The involvement of the diverticula, a synapomorphy for Itunina, in protrusion and expansion of hairpencils by male Lycorea halia (Hübner, 1816) is demonstrated for the first time. They facilitate maintaining the haemolymph pressure necessary to keep the hairpencils everted. The diverticula are curved hook-like lobes, open to the body cavity and densely filled with tracheae and threads made by units of two staggered cells surrounding a central extracellular fibril bundle. Such complex structures, apparently metabolically active, have not been reported for insects previously and might indicate additional functions, but their functional role(s) remains a puzzle. When a male emerges from pupa, the diverticula are not yet formed; this happens only during the first protrusion of the hairpencils.

Under pressure: force resistance measurements in box mites (Actinotrichida, Oribatida)

BackgroundMechanical defenses are very common and diverse in prey species, for example in oribatid mites. Here, the probably most complex form of morphological defense is known as ptychoidy, that enables the animals to completely retract the appendages into a secondary cavity and encapsulate themselves. The two groups of ptychoid mites constituting the Ptyctima, i.e. Euphthiracaroidea and Phthiracaroidea, have a hardened cuticle and are well protected against similar sized predators. Euphthiracaroidea additionally feature predator-repelling secretions. Since both taxa evolved within the glandulate group of Oribatida, the question remains why Phthiracaroidea lost this additional protection. In earlier predation bioassays, chemically disarmed specimens of Euphthiracaroidea were cracked by the staphylinid beetle Othius punctulatus, whereas equally sized specimens of Phthiracaroidea survived. We thus hypothesized that Phthiracaroidea can withstand significantly more force than Euphthiracaroidea and that the specific body form in each group is key in understanding the loss of chemical defense in Phthiracaroidea. To measure force resistance, we adapted the principle of machines applying compressive forces for very small animals and tested the two ptyctimous taxa as well as the soft-bodied mite Archegozetes longisetosus.ResultsSome Phthiracaroidea individuals sustained about 560,000 times their body weight. Their mean resistance was about three times higher, and their mean breaking point in relation to body weight nearly two times higher than Euphthiracaroidea individuals. The breaking point increased with body weight and differed significantly between the two taxa. Across taxa, the absolute force resistance increased sublinearly (with a 0.781 power term) with the animal’s body weight. Force resistance of A. longisetosus was inferior in all tests (about half that of Euphthiracaroidea after accounting for body weight). As an important determinant of mechanical resistance in ptychoid mites, the individuals’ cuticle thickness increased sublinearly with body diameter and body mass as well and did not differ significantly between the taxa.ConclusionWe showed the feasibility of the force resistance measurement method, and our results were consistent with the hypothesis that Phthiracaroidea compensated its lack of chemical secretions by a heavier mechanical resistance based on a different body form and associated build-up of hemolymph pressure (defensive trade-off).

Water vapor absorption allows for volume expansion during molting in Armadillidiumvulgare and Porcelliodilatatus (Crustacea, Isopoda, Oniscidea).

Arthropods require periodic molting in order to grow which presents a number of challenges to terrestrial taxa. Following ecdysis, the pliant new cuticle is susceptible to buckling under gravity and requires elevated hydrostatic pressure for support. Terrestrial species also require a mechanism of volume expansion and stretching of the integument prior to sclerotization, a need that is readily met in aquatic arthropods by drinking. Options for land arthropods include drinking of dew, swallowing of air, or using muscular contractions to inflate air sacs in tracheate taxa. In this study we tested the hypothesis that crinochete terrestrial isopods (Isopoda: Oniscidea: Crinocheta) exploit their capacity for active water vapor absorption (WVA) to increase volume during molting. Two crinochete species, Armadillidiumvulgare and Porcelliodilatatus, were studied and compared with the non-absorbing species Ligidiumlapetum (Oniscidea: Ligiamorpha). Pre-molting animals were identified by sternal CaCO3 deposits and exposed to 100% or 97% relative humidity (RH). Mass-changes were monitored by daily weighing and the timing of the posterior and anterior ecdyses was used to categorize time (days premolt and days post-molt) over the molt cycle. In each treatment RH, A.vulgare and P.dilatatus showed a progressive mass increase from 5 days premolt until the posterior or anterior ecdysis, followed abruptly by period of mass-loss lasting 3–4 days post-molt. The fact that the initial mass-gain is seen in 97 % RH, a humidity below the water activity of the hemolymph, confirms the role of WVA. Similarly, since the post-molt mass-loss is seen in 100 % RH, this must be due to active expulsion of water, possibly via maxillary urine. Concurrent changes in hemolymph osmolality were monitored in a separate batch of A.vulgare and show sustained osmolality during premolt and an abrupt decrease between the anterior and posterior ecdysis. These patterns indicate a mobilization of sequestered electrolytes during premolt, and a loss of electrolytes during the post-molt mass-loss, amounting to approximately 8.6 % of total hemolymph solutes. WVA, in conjunction with pulses of elevated hemolymph pressure, provides an efficient mechanism of pre-molt volume expansion prior to and during the biphasic molt in these species. Premolt Ligidiumlapetum exposed to same treatments failed to molt successfully and no premolt animals survived to day 3 (72 h) even in 100 % RH. The apparent dependence of this species on liquid water for successful molting could explain its obligatory association with riparian fringe habitats.

Influence of hydrodynamic pressure and vein strength on the super-elasticity of honeybee wings

The wings of honeybees (Apis mellifera L.) usually produce bending and torsional deformations during flapping wing movement. These deformations endow honeybees with perfect aerodynamic control to escape predators and exploit scattered resources. However, the mechanisms by which honeybee wings recover from large deformations are unclear. This study demonstrates that honeybee wings are super-elastic that they can recover rapidly from one extreme contorted state to their original position. A comparative experiment is conducted to evaluate the difference in super-elastic recovery between attached and detached wings. Results show that the structural stiffness of wings is affected by the reticulate vein and the haemolymph pressure generated by the blood circulation. Further analysis indicates that the haemolymph pressure can increase the stiffness of honeybee wings, especially that of the subcostal veins. This phenomenon shortens the recovery time of wing deflection behaviour.

Effect of erythritol formulation on the mortality, fecundity and physiological excretion in Drosophila suzukii

Previously, we studied various combinations of non-nutritive sugars including erythritol and erythrose having a potentially insecticidal effect on Drosophila suzukii. The study suggested two potential physiological changes causing fly mortality: 1) starvation from the feeding of non-metabolizable erythritol and erythrose; 2) abnormal osmotic pressure increased in the hemolymph with erythritol transported from the midgut. In the present study, sucrose and erythritol were applied to blueberries and effects of these combinations on fly mortality and fecundity were monitored in the lab and greenhouse. In the lab, two sucrose/erythritol formulations (0.5M sucrose/2M erythritol, 1M sucrose/2M erythritol) resulted in the highest mortality and the lowest fecundity among D. suzukii adults. Two formulations, therefore, were selected for further evaluation with blueberry bushes and fruits in the greenhouse; fly survival with 0.5M sucrose/2M erythritol was significantly lower than 1M sucrose/2M erythritol for 7days. Unlike the smaller container, mortality occurred faster in the greenhouse probably because flies moved more in the bigger cage accelerating the exhaustion of energy reserves in the body. We examined presence of erythritol in the hemolymph and frass to determine the nutritional metabolism and absorption of erythritol in D. suzukii. Unlike sucrose, a large amount of erythritol was observed in the hemolymph of the fly that ingested 0.5M sucrose/0.5M erythritol. Erythritol was also found in the frass of the same fly. The results imply that erythritol might be directly transported from the midgut without being metabolized and stored, but is accumulated in the hemolymph which in turn elevates the osmotic pressure in the fly hemolymph. For practical application, the sucrose/erythritol combination would be more effective than erythritol alone because the combination tastes sweeter to elicit more feeding. This erythritol formulation can be a potential insecticide used alone or as a delivery agent combined with conventional or biological insecticides to enhance their efficacy.

NOTES ON THE BIOLOGY OF Oncideres bouchardii BATES (Coleóptera: Cerambycidae), A TRUPILLO TWIG GIRDTER FROM SANTA MARTA, COLOMBIA

Aspects of the biology of Oncideres bouchardii Bates from Santa Marta, Colombia are described. Imagines of this twig girdler appear shortly after the rainy season during the months of November to February, ringing twigs and branches of trupillo trees, Prosopis juliflora (L.) and aromo bushes, Acacia fameÿiana Willd. Observations showed that this activity is only performed by the females, which commonly copulate at the same time. After girdling the twig, the females lay their white eggs one by one into especially prepared egg cavities that are sealed after oviposition. There is evidence that locating the entrance to the future egg chamber with the ovipositor is greatly facilitated by means of a marking on the bark made with the ovipositor while making the entrance with the mandibles. The eggs are deposited by means of the telescopic and inflatable ovipositor which is inserted deeply under the bark through hemolymph pressure and acts to force open the egg chamber between bark and wood. After approximately a week the larvae hatch and soon dig into the drying twig. The lifecycle of Oncideres bouchardii lasts one year, the longest part of it belonging to the larval stage. Most imagines live less than a week or two, although a few survive as long as 45 days. In places where much destruction of the trees is caused by this twig girdler, it is recommended to collect and burn the girdled twigs or branches that contain the woodboring larvae or eggs of the species.