'Drone-Netting' for Sampling Live Insects.

Drones have become valuable tools for biodiversity studies by providing aerial photographs; however, for most entomological studies, images, in particular those taken remotely, are usually insufficient; rather sampling of specimens is required. We equipped a cheap off-the-shelf drone with a net bag, flew it over the ground, sweeping the vegetation, and sampled adult and larval insects as well as spiders. 'Drone-netting' proved to be a versatile method for general insect sampling, particularly in inaccessible terrains. It is time- and cost-effective, minimally invasive, and adaptable for many research tasks in entomofaunistics; it shows a degree of representativeness similar to hand-netting, and caught specimens stay alive and can be released if not needed.

'Crystal Macrosetae': Novel Scales and Bristles in Male Arctiine Moths (Lepidoptera: Erebidae: Arctiinae) Filled with Crystallizing Material.

Scales, exoskeletal features characteristic of the Lepidoptera, occur in enormous structural and functional diversity. They cover the wing membranes and other body parts and give butterflies and moths their often stunning appearance. Generally, the patterns made by scales are visual signals for intra- and interspecific communication. In males, scales and/or bristles also make up the androconial organs, which emit volatile signals during courtship. Here, a structurally and putative functionally novel type of scales and bristles is reported: 'crystal macrosetae'. These lack trabeculae and windows, are made up by a very thin and flexible envelope only and contain crystallizing material. In 'crystal scales', there is a flat surface ornamentation of modified ridges, while 'crystal bristles' often show large protrusions. Crystal macrosetae usually cannot be reliably recognized without destruction. Apparently, they serve as containers for large amounts of material that is viscous in living moths, highly hygroscopic, crystallizes when specimens dry up, and can be visualized by scanning electron microscopy. Crystal macrosetae occur in males only, always associated with or making up androconial organs located on various parts of the body, and have numerous forms with diverse surface ornamentation across many species and genera. The newly identified structures and the discovery of crystallizing material in scales and bristles raise many questions and could shed new light on ontogenetic development of macrosetae, and on the biology and physiology as well as the evolution and systematics of Arctiinae. There is evidence that crystal macrosetae occur in other moths too.

Twittering Pupae of Papilionid and Nymphalid Butterflies (Lepidoptera): Novel Structures and Sounds.

Pupae of numerous Papilionidae and Nymphalidae produce twitter sounds when wriggling in response to mechanical stimulation. The structural basis comprises distinct pairs of sound-producing organs (SPOs) located at intersegmental membranes of the abdomen. They differ-as the twitters do-in sampled taxa of Papilioninae, Epicaliini, and Heliconiini. The opposing sculptured cuticular sound plates (SPs) of each SPO appear structurally the same but are actually mirror-images of each other. Results suggest that sounds are not generated by stridulation (friction of a file and a scraper) but when these inversely sculptured and interlocking surfaces separate during pupal wriggling, representing a stick-slip mechanism. Twitter sounds comprise series of short broadband pulses with the main energy in the frequency range 3-13 kHz; they can be heard by humans but extend into ultrasonic frequencies up to 100 kHz.

Pro-toxic 1,2-Dehydropyrrolizidine Alkaloid Esters, Including Unprecedented 10-Membered Macrocyclic Diesters, in the Medicinally-used Alafia cf. caudata and Amphineurion marginatum (Apocynaceae: Apocynoideae: Nerieae and Apocyneae).

INTRODUCTION: Within the Apocynoideae (Apocynaceae) pro-toxic dehydropyrrolizidine alkaloids have been reported only in Echiteae. However, attraction of pyrrolizidine alkaloid-pharmacophagous insects suggested their presence in Alafia cf. caudata Stapf (Nerieae: Alafiinae) and Amphineurion marginatum (Roxb.) D.J. Middleton (Apocyneae: Amphineuriinae), both used as medicinal plants. OBJECTIVE: To confirm the presence of dehydropyrrolizidine alkaloids in Alafia cf. caudata and Amphineurion marginatum and identify their structures. METHODS: Methanol extracts of air-dried roots, stems and leaves of non-flowering plants were analysed using HPLC-ESI(+)MS and MS/MS or collision-induced dissociation MS in low and/or high resolution modes. Pyrrolizidine alkaloids were tentatively identified based on the mass spectrometry data. Solid phase extraction combined with semi-preparative HPLC were used to isolate major alkaloids. Structures were elucidated using NMR spectroscopy. RESULTS: Monoesters of retronecine with senecioic, hydroxysenecioic or syringic acids were identified in roots of Alafia cf. caudata. Two unprecedented 10-membered macrocyclic dehydropyrrolizidine alkaloid diesters were isolated from roots of Amphineurion marginatum. Pyrrolizidine alkaloids were detected in root and leaf material of Alafia cf. caudata at 0.34 and 0.01% dry weight (DW), and 0.13, 0.02 and 0.09% DW in root, leaf and stem material of Amphineurion marginatum. CONCLUSIONS: The presence of pro-toxic dehydropyrrolizidine alkaloids suggests that medical preparations of these plants pose potential health risks to consumers. Dehydropyrrolizidine alkaloids are evidently more widespread in Apocynoideae than previously assumed, and it would seem rewarding to study other members of this family for the presence of pyrrolizidines, dehydropyrrolizidines and dihydropyrrolizines. Copyright © 2016 John Wiley & Sons, Ltd.

Hepatotoxic pyrrolizidine alkaloids in pollen and drying-related implications for commercial processing of bee pollen.

Using HPLC-ESI-MS, several saturated and 1,2-dehydropyrrolizidine alkaloids were detected, mainly as their N-oxides, in fresh pollen collected from flowers of the pyrrolizidine alkaloid-producing plants Echium vulgare, E. plantagineum, Senecio jacobaea, S. ovatus, and Eupatorium cannabinum, and/or pollen loads from bees (bee pollen) that foraged on those plants. A major alkaloidal metabolite in S. ovatus was tentatively identified, using its mass spectrometric data and biogenic considerations, as the previously unreported, saturated alkaloid, 2-hydroxysarracine. Heating had very little effect on the 1,2-dehydropyrrolizidine alkaloids and their N-oxides from a variety of sources. Considered in conjunction with international concerns about the adverse effects of these alkaloids, the results strongly indicate a need for monitoring pollen supplies intended for human consumption, at least until conditions for processing and/or selection are clearly defined such as to significantly reduce the hepatotoxic (and potentially carcinogenic and genotoxic) pyrrolizidine alkaloid content of bee pollen.