Ivar Sibul

Sub-lethal effects of the neurotoxic pyrethroid insecticide Fastac 50EC on the general motor and locomotor activities of the non-targeted beneficial carabid beetle Platynus assimilis (Coleoptera: Carabidae).

BACKGROUND Sub-lethal effects of pesticides on behavioural endpoints are poorly studied in carabids (Coleoptera: Carabidae) though changes in behaviour caused by chemical stress may affect populations of these non-targeted beneficial insects. General motor activity and locomotion are inherent in many behavioural patterns, and changes in these activities that result from xenobiotic influence mirror an integrated response of the insect to pesticides. Influence of pyrethroid insecticides over a wide range of sub-lethal doses on the motor activities of carabids still remains unclear. RESULTS Video tracking of Platynus assimilis showed that brief exposure to alpha-cypermethrin at sub-lethal concentrations ranged from 0.01 to 100 mg L(-1) caused initial short-term (< 2 h) locomotor hyperactivity followed by a long-term (>24 h) locomotor hypo-activity. In addition, significant short- and long-term concentration and time-dependent changes occurred in general motor activity patterns and rates. CONCLUSION Conspicuous changes in motor activity of Platynus assimilis beetles treated at alpha-cypermethrin concentrations up to 75,000-fold lower than maximum field recommended concentration (MFRC) suggest that many, basic fitness-related behaviours might be severely injured as well. These changes may negatively affect carabid populations in agro-ecosystems. Long-term hypo-activity could directly contribute to decreased trap captures of carabids frequently observed after insecticide application in the field.

Monitoring of gas exchange cycles and ventilatory movements in the pine weevil Hylobius abietis : respiratory failures evoked by a botanical insecticide

The large pine weevil, Hylobius abietis (L.) (Coleoptera: Curculionidae), is the most important insect pest of young coniferous plants. The implementation of new control methods requires not only a profound knowledge of the ecology and behaviour of the pest, but particularly of its physiology. Standard metabolic rate (SMR) and discontinuous gas exchange cycles (DGCs) were recorded in parallel with abdominal ventilation movements in adults of H. abietis using a differential electrolytic respirometer‐actograph. Quiescent weevils displayed DGCs of the constriction, flutter, and ventilation phases of the CFV type, while bursts of carbon dioxide were always accompanied by abdominal pumping movements, i.e., muscular ventilation in the closed subelytral cavity (SEC). In some beetles the C phase was absent and thus (C)FV cycles were recorded. In addition, at the beginning and often at the end of a burst, the SEC was rhythmically opened and closed by movements of the last abdominal segments. Continuous pumping movements and an absence of DGCs were signs of stress imposed by handling or by a new environment, even if the beetle was not moving. All individuals showed clear DGCs after recovering from handling and apparatus stress lasting 2–3 h. The results show that in the monitoring of DGCs, it is essential to determine whether they are of the constriction, flutter, and open phases (CFO), or the CFV subtype of the constriction, flutter, and burst (CFB) cycles. Use of our simple closed‐system respirometer enables non‐invasive simultaneous recording of SMR, oxygen uptake, DGCs, and active ventilation in H. abietis and other beetles.

Sensitivity of antennal gustatory receptor neurones to aphid honeydew sugars in the carabid Anchomenus dorsalis

Using electrophysiology, the stimulating effect of 13 sugars and three sugar alcohols (each at a concentration of 100 mm) to antennal gustatory receptor neurones (GRNs) is tested in the carabid beetle Anchomenus dorsalis (Pontoppidan, 1763) (Coleoptera, Carabidae). Maltose, sucrose, glucose and raffinose are the most stimulating sugars for the sugar‐sensitive neurone (SuN), evoking 6.7–18.6 spikes s−1 in fed insects, whereas the others had little or no effect. The firing rate of the antennal GRNs is not affected by any of the tested sugar alcohols, dulcitol, inositol and sorbitol. Additionally, concentration/response curves for sucrose and maltose are obtained in the range 0.01–100 mm. The responses of beetles starved for 96 h to this range of sucrose are two‐ to three‐fold higher compared with those of fed beetles. The presence of a terminal α‐glucose unit is an important feature of the molecular structure determining the stimulating properties of the two disaccharides, maltose and sucrose, as well as glucose. The other monosaccharide unit of the molecule is also of great importance in determining the stimulating properties of various disaccharides. The sensitivity of the SuN to the four most prevalent aphid honeydew sugars suggests that A. dorsalis uses these chemicals as sensory cues when searching for aphids as prey.

Responses of the antennal bimodal hygroreceptor neurons to innocuous and noxious high temperatures in the carabid beetle, Pterostichus oblongopunctatus

Electrophysiological responses of thermo- and hygroreceptor neurons from antennal dome-shaped sensilla of the carabid beetle Pterostichus oblongopunctatus to different levels of steady temperature ranging from 20 to 35°C and rapid step-changes in it were measured and analysed at both constant relative and absolute ambient air humidity conditions. It appeared that both hygroreceptor neurons respond to temperature which means that they are bimodal. For the first time in arthropods, the ability of antennal dry and moist neurons to produce high temperature induced spike bursts is documented. Burstiness of the spike trains is temperature dependent and increases with temperature increase. Threshold temperatures at which the two neurons switch from regular spiking to spike bursting are lower compared to that of the cold neuron, differ and approximately coincide with the upper limit of preferred temperatures of the species. We emphasise that, in contrast to various sensory systems studied, the hygroreceptor neurons of P. oblongopunctatus have stable and continuous burst trains, no temporal information is encoded in the timing of the bursts. We hypothesise that temperature dependent spike bursts produced by the antennal thermo- and hygroreceptor neurons may be responsible for detection of noxious high temperatures important in behavioural thermoregulation of carabid beetles.

Gas exchange patterns of Pterostichus niger (Carabidae) in dry and moist air

Gas exchange patterns of adult male Pterostichus niger Schaller after hydration (i.e. given access to food and water) are compared in dry air [5–7% relative humidity (RH)] and moist air (90–97% RH) by means of flow‐through CO2 respirometry combined with infrared probe actography. Of thirty beetles examined, slightly more than 50% showed continuous gas exchange and are not considered further. Of the remaining beetles, the majority (approximately 71%) display a pattern of cyclic gas exchange in both dry and moist air (i.e. CO2 gas is released in bursts, with a low level of CO2 release during the interburst periods). A minority of the beetles (four out of 30) are found to exhibit discontinuous gas exchange in both dry and moist air; this is characterized by three clearly separated states of the spiracles: closed (C), flutter (F) and open (O) phases. The pattern of cyclic gas exchange is associated with weak abdominal pulsations. After switching from moist to dry air, a small modulation of the discontinuous gas exchange cycles (maximum mean CO2 production rate) occurs, providing no clear support for the hygric theory of discontinuous gas exchange in this species (i.e. that it serves to restrict respiratory water loss).

Behavioural effects of the neonicotinoid insecticide thiamethoxam on the predatory insect Platynus assimilis

Little information is available regarding sublethal effects of neonicotinoids on insect predators, many of which perform important roles in ecosystem functioning and biocontrol. In this study, dose-dependent sublethal effects of a dietary administered neonicotinoid insecticide thiamethoxam on two basic behaviours, locomotion and feeding, were quantified in the carabid Platynus assimilis (Coleoptera, Carabidae) using automated video-tracking and weighing of consumed food, respectively. Acute toxicity tests showed that, when orally administered, the LD50 of thiamethoxam for P. assimilis beetles was 114.5 ng/g. Thiamethoxam at 108.1 ng/g caused a short-term locomotor hyperactivity within several hours of treatment. Next day after exposure to the insecticide, all the beetles were in a state of locomotor hypoactivity independent of the administered dose ranging from 1.1 to 108.1 ng/g. Reduction in clean food consumption rate (CFCR) is another altered behavioural endpoint of poisoned insect predators as first demonstrated in this study. On the first day of thiamethoxam administration, a remarkable reduction in feeding only occurred in beetles treated at 108.1 ng/g but on the next day, this negative effect appeared even at doses ten to a hundred-fold lower. Recovery from locomotion abnormalities and reduced feeding took several days. Both locomotor activity and CFCR are sensitive and valuable ecotoxicological biomarkers of carabids which should be taken into account in Integrated Pest Management programs where optimal combination of reduced insecticide use and biological control by predatory insects is crucial to achieve best results.

The large pine weevil Hylobius abietis (L.) as a potential vector of the pathogenic fungus Diplodia sapinea (Fr.) Fuckel

Diplodia sapinea, an important pathogen of various conifer species, was recently recorded in the northern Baltic region. The pathogen can disperse over short distances via rain or wind, whereas long range dispersal occurs via movement of contaminated plant material and seeds by humans, as well as by insects. Hylobius abietis is one of the most important forest pests over large areas of Europe. Adult weevils feed on the bark of seedlings of young conifers, causing injury and often death. Weevils were collected from fresh clear‐cut areas and near the symptomatic conifer stands to analyze the presence of D. sapinea. Entire weevils collected from natural environments were crushed without surface washing. The identity of D. sapinea and the fungal DNA extraction from the insects was confirmed by species‐specific polymerase chain reaction priming. Eight H. abietis individuals were determined to be infected with D. sapinea. In a laboratory experiment, pine branches were infected with D. sapinea and were fed to adult H. abietis. The results show that the pathogen survived in the digestive tract of H. abietis in the laboratory experiment, and also that the isolation of fungus from the faeces was successful. In addition, the results demonstrate that the large pine weevil may be a potential vector of pathogenic fungus D. sapinea.

The opening-closing rhythms of the subelytral cavity associated with gas exchange patterns in diapausing Colorado potato beetle, Leptinotarsa decemlineata.

ABSTRACT The opening–closing rhythms of the subelytral cavity and associated gas exchange patterns were monitored in diapausing Leptinotarsa decemlineata beetles. Measurements were made by means of a flow-through CO2 analyser and a coulometric respirometer. Under the elytra of these beetles there is a more or less tightly enclosed space, the subelytral cavity (SEC). When the cavity was tightly closed, air pressure inside was sub-atmospheric, as a result of oxygen uptake into the tracheae by the beetle. In about half of the beetles, regular opening–closing rhythms of the SEC were observed visually and also recorded; these beetles displayed a discontinuous gas exchange pattern. The SEC opened at the start of the CO2 burst and was immediately closed. On opening, a rapid passive suction inflow of atmospheric air into the SEC occurred, recorded coulometrically as a sharp upward peak. As the CO2 burst lasted beyond the closure of the SEC, we suggest that most of the CO2 was expelled through the mesothoracic spiracles. In the remaining beetles, the SEC was continually semi-open, and cyclic gas exchange was exhibited. The locking mechanisms and structures between the elytra and between the elytra and the body were examined under a stereomicroscope and by means of microphotography. We conclude that at least some of the L. decemlineata diapausing beetles were able to close their subelytral cavity tightly, and that the cavity then served as a water-saving device. Summary: Colorado potato beetles are able to close their subelytral cavity tightly and it then serves as a water-saving device.

Variability in metabolic rate and gas exchange patterns of the Colorado potato beetle of winter and prolonged diapauses: Gas exchange patterns of Colorado potato beetle

We compare standard metabolic rate (VCO2) and gas exchange patterns in the Colorado potato beetle (Leptinotarsa decemlineata Say) in winter diapause (i.e. lasting only one overwintering period) with those of beetles in prolonged diapause (i.e. diapause lasting 2 or 3 years). The length of diapause is estimated by the behaviour of the beetles: burrowing into soil as a sign of the beginning of diapause and emergence from the soil as the ending of diapause. Measurement with a flow‐through carbon dioxide (CO2) infrared gas analyzer reveals that most beetles in winter or prolonged diapause display distinct discontinuous gas exchange cycles at 23 °C. Beetles with cyclic gas exchange and continuous breathing do not survive the winter. Beetles in prolonged diapause are characterized by a three‐ to five‐fold lower standard metabolic rates, longer discontinuous gas exchange periods and shorter CO2 releases (open phase) than those in winter diapause.

Gas exchange pattern in the two‐spot ladybird beetle, Adalia bipunctata, differs in dry vs. moist air

Gas exchange patterns in the ladybird beetle, Adalia bipunctata (L.) (Coleoptera: Coccinellidae), were investigated using an infrared gaseous analyser (IRGA) and a coulometric O2 respirometer (manometric–volumetric system). Before testing, the beetles were kept either in dry (dehydrated) or moist (hydrated) conditions for 1 day. Their subsequent gas exchange patterns did not depend on their state of humidity but rather were controlled by the humidity of the insect chamber during gas exchange measurement. If this chamber contained dry air, the beetles exhibited CO2 release by burst, which we interpreted as cyclic gas exchange (CGE) with inter‐burst periods, but if the chamber was switched to contain moist air, then cyclic CO2 release was soon abandoned and a pattern of continuous gas exchange appeared. Measurements with the coulometric respirometer in moist air showed that continuous gas exchange was often associated with weak abdominal pulsations, which we interpreted as active ventilation. Their metabolic rate was lower during gas exchange cycles than during continuous gas exchange. We revealed that in the ladybird beetle metabolic rate increased in moist air when the gas exchange pattern transitioned from cyclic to continuous.