An X-ray micro-computer tomography study of the Malpighian tubules of the Blue Bottle Blow Fly (Calliphora vomitoria) Diptera: Calliphoridae.

Malpighian tubules are the insect equivalent of mammalian kidneys and normally drain into the gut at the junction between the mid and hind gut. The Malpighian tubules of the fruit fly Drosophila melanogaster are increasingly being used as a model for studying human renal tract development, histology, nephrolithiasis and urolithiasis. In the present study we report when using X-ray micro-computer tomography techniques, the larval, intrapuparial and adult stages of the larger Calliphora vomitoria can contain large amounts of calcium-rich concretions which are tightly packed in the lumen of both anterior Malpighian tubules. We show that it is feasible to utilise these calcium-rich concretions as a form of marking agent to delineate the various developmental stages of the Malpighian tubules including the crucial phase when the Malpighian tubules reconnect with the hind gut. In the majority of cases during the intrapuparial period the ureters of the Malpighian tubules did not start to re-canalise and thus reconnect with the developing hind gut until the 7th day of the 10-11 day. Just prior to ecdysis, virtually all the radio-opaque concretions in the Malpighian tubules had emptied into the hind gut and had then been completely excreted by the time the imago emerged from its puparium. In contrast, we show that in flies developing from larvae previously stained by ingesting Rhodamine B, a known substrate for both the Multi Xenobiotic Resistance and Multi Drug Resistant membrane transport systems, the efficiency with which these calcium-rich concretions are excreted by the imago as it emerges from its intrapuparial period can be significantly impaired. Therefore, it might be useful to include C. vomitoria as a model when studying renal tract development and urolithiasis using X-ray micro-computer tomography.

Radio-Frequency Electromagnetic Field Exposure of Western Honey Bees.

Radio-frequency electromagnetic fields (RF-EMFs) can be absorbed in all living organisms, including Western Honey Bees (Apis Mellifera). This is an ecologically and economically important global insect species that is continuously exposed to environmental RF-EMFs. This exposure is studied numerically and experimentally in this manuscript. To this aim, numerical simulations using honey bee models, obtained using micro-CT scanning, were implemented to determine RF absorbed power as a function of frequency in the 0.6 to 120 GHz range. Five different models of honey bees were obtained and simulated: two workers, a drone, a larva, and a queen. The simulations were combined with in-situ measurements of environmental RF-EMF exposure near beehives in Belgium in order to estimate realistic exposure and absorbed power values for honey bees. Our analysis shows that a relatively small shift of 10% of environmental incident power density from frequencies below 3 GHz to higher frequencies will lead to a relative increase in absorbed power of a factor higher than 3.

Exposure of Insects to Radio-Frequency Electromagnetic Fields from 2 to 120 GHz.

Insects are continually exposed to Radio-Frequency (RF) electromagnetic fields at different frequencies. The range of frequencies used for wireless telecommunication systems will increase in the near future from below 6 GHz (2 G, 3 G, 4 G, and WiFi) to frequencies up to 120 GHz (5 G). This paper is the first to report the absorbed RF electromagnetic power in four different types of insects as a function of frequency from 2 GHz to 120 GHz. A set of insect models was obtained using novel Micro-CT (computer tomography) imaging. These models were used for the first time in finite-difference time-domain electromagnetic simulations. All insects showed a dependence of the absorbed power on the frequency. All insects showed a general increase in absorbed RF power at and above 6 GHz, in comparison to the absorbed RF power below 6 GHz. Our simulations showed that a shift of 10% of the incident power density to frequencies above 6 GHz would lead to an increase in absorbed power between 3-370%.

Using diagnostic radioentomology for non-invasive observations of colonies of the bumblebee, Bombus terrestris.

Bumblebees have been the focus of a broad range of scientific research due to their behavior, social life, and a number of other intriguing traits. Current methods for examining their nest structure, such as natal cells and contents of storage cells, are destructive in nature because the cells need to be opened for physical inspections. This research describes how the internal structures of the artificial nests of the bumblebee Bombus terrestris L. (Hymentoptera: Apidae) were non-invasively viewed and assessed by using diagnostic radioentomology. For the first time, B. terrestris nest structures, and their contents such as larvae, pupae and eggs, were non-invaseively viewed and assessed. This technique will enable future experiments to take morphological measurements of egg, larval, and pupal development over time. Moreover, combining these measurements with measures of food-storage will provide a good assessment of colony health. The method will also allow tracking of individually marked adults, to monitor their behaviour and help gain a better understanding of the processes involved in the global declines of B. terrestris, which will in turn promote better management of these valuable pollinators.

Imaging live bee brains using minimally-invasive diagnostic radioentomology.

The sensitivity of the honey bee, Apis mellifera L. (Hymeonoptera: Apidae), brain volume and density to behavior (plasticity) makes it a great model for exploring the interactions between experience, behavior, and brain structure. Plasticity in the adult bee brain has been demonstrated in previous experiments. This experiment was conducted to identify the potentials and limitations of MicroCT (micro computed tomograpy) scanning "live" bees as a more comprehensive, non-invasive method for brain morphology and physiology. Bench-top and synchrotron MicroCT were used to scan live bees. For improved tissue differentiation, bees were fed and injected with radiographic contrast. Images of optic lobes, ocelli, antennal lobes, and mushroom bodies were visualized in 2D and 3D rendering modes. Scanning of live bees (for the first time) enabled minimally-invasive imaging of physiological processes such as passage of contrast from gut to haemolymph, and preliminary brain perfusion studies. The use of microCT scanning for studying insects (collectively termed 'diagnostic radioentomology', or DR) is increasing. Our results indicate that it is feasible to observe plasticity of the honey bee brain in vivo using diagnostic radioentomology, and that progressive, real-time observations of these changes can be followed in individual live bees. Limitations of live bee scanning, such as movement errors and poor tissue differentiation, were identified; however, there is great potential for in-vivo, non-invasive diagnostic radioentomology imaging of the honey bee for brain morphology and physiology.

The alternative Pharaoh approach: stingless bees mummify beetle parasites alive.

Workers from social insect colonies use different defence strategies to combat invaders. Nevertheless, some parasitic species are able to bypass colony defences. In particular, some beetle nest invaders cannot be killed or removed by workers of social bees, thus creating the need for alternative social defence strategies to ensure colony survival. Here we show, using diagnostic radioentomology, that stingless bee workers (Trigona carbonaria) immediately mummify invading adult small hive beetles (Aethina tumida) alive by coating them with a mixture of resin, wax and mud, thereby preventing severe damage to the colony. In sharp contrast to the responses of honeybee and bumblebee colonies, the rapid live mummification strategy of T. carbonaria effectively prevents beetle advancements and removes their ability to reproduce. The convergent evolution of mummification in stingless bees and encapsulation in honeybees is another striking example of co-evolution between insect societies and their parasites.