Bio-analysis of insects is increasingly dependent on highly sensitive methods that require high quality biological material, such as hemolymph. However, it is difficult to collect fresh and uncontaminated hemolymph from adult bees since they are very active and have the potential to sting, and because hemolymph is rapidly melanized. Here we aimed to develop and test a quick and easy method for sterile and contamination-free hemolymph sampling from adult Apidae. Our novel antennae method for hemolymph sampling (AMHS), entailed the detachment of an antenna, followed by application of delicate pressure to the bee's abdomen. This resulted in the appearance of a drop of hemolymph at the base of the detached antenna, which was then aspirated using an automatic pipetter. Larger insect size corresponded to easier and faster hemolymph sampling, and to a greater sample volume. We obtained 80–100 μL of sterile non-melanized hemolymph in 1 minute from one Bombus terrestris worker, in 6 minutes from 10 Apis mellifera workers, and in 15 minutes from 18 Apis cerana workers (+/−0.5 minutes). Compared to the most popular method of hemolymph collection, in which hemolymph is sampled by puncturing the dorsal sinus of the thorax with a capillary (TCHS), significantly fewer bees were required to collect 80–100 μL hemolymph using our novel AMHS method. Moreover, the time required for hemolymph collection was significantly shorter using the AMHS compared to the TCHS, which protects the acquired hemolymph against melanization, thus providing the highest quality material for biological analysis.
Many Viburnum species are popular ornamental shrubs and, simultaneously, highly valued medicinal plants as a source of many bioactive compounds, including antioxidants. Viburnum bark, flowers, and fruits are widely used in traditional and folk medicine, and the fruits of some species are used as cooking ingredients. The knowledge of the microstructure of Viburnum fruits and the accumulation sites of bioactive substances in these organs is rather poor. Comparative analyses of the microstructure of ripe Viburnum opulus and Viburnum lantana drupes were carried out using light, scanning, and transmission electron microscopes. The location of various groups of metabolites in the fruits of both species was determined with the use of histochemical tests and fluorescence microscopy. Additionally, the major antioxidants, i.e. carotenoids, polyphenols, and flavonoids, were quantified and a number of morphometric traits of the drupes were presented. The V. opulus and V. lantana fruits were found to differ in some morphological traits and in many characteristics of the pericarp anatomy and ultrastructure. It was shown that the Viburnum fruits contained lipids and lipid compounds (carotenoids, essential oils, steroids, and saponins), polyphenols (tannins, flavonoids, and anthocyanins), pectins, and proteins. The fruits of V. opulus contained greater quantities of carotenoids, polyphenols, flavonoids, steroids, and pectins than the V. lantana drupes, whereas the latter were characterised by higher contents of essential oils, saponins, and proteins. The metabolites were located in different pericarp layers, but the greatest amounts were identified in the drupe skin.
A b s t r a c tThe anatomical structure and the distribution of arabinogalactan proteins were investigated in the ovule of an energetic plant Sida hermaphrodita (L.) Rusby after pollination. Material was collected from an experimental field of the University of Life Sciences in Lublin (Felin). After embedding for immunoflurescence, semi-thin sections were stained with toluidine blue or incubated with JIM 13 or MAC 207 monoclonal antibodies for immunolabeling of arabinogalactan proteins. The structure of the ovule showed some differences in comparison to other representatives of the Malvaceae family. Conversely, the distribution of arabinogalactan proteins did not differ significantly from their distribution in the ovules of other angiosperms at the investigated stage of development.
Pollen of the Antarctic plants Colobanthus quitensis and Deschampsia antarctica and its representation in moss polstersPyłek antarktycznych roślin Colobanthus quitensis i Deschampsia antarctica i jego reprezentacja w próbkach mchów SUMMARY Formation, structure, and representation in moss polsters of pollen grains of Colobanthus quitensis (Kunth) Bartl. and Deschampsia antarctica Desv. (the only two native flowering plants growing in Antarctica) were investigated by means of light microscopy. Microsporogenesis and male gametogenesis of both investigated plant species proceded in the way typical of other angiosperms. C. quitensis forms spherical, two-nuclear pollen grains enveloped by the thick polyporate sporoderm. Pollen grains of D. antarctica are three-nuclear, and their sporoderm contains one aperture. Both investigated species form chasmogamic and cleistogamic flowers. In moss samples, collected according to Pollen Monitoring Programme method, the local element -Poaceae and Colobanthus prevailed. The Poaceae pollen type includes mostly Deschampsia, but other grains, such as Poa annua, which was reported to grow in the vicinity of H. Arctowski Station, cannot be excluded. Among long-distance elements, Nothofagus pollen prevailed. One to seven pollen grains of this tree were found in each sample, which had been transported by strong westerly winds from southern South America. All the examined spectra also contained numerous fungal spores and other unidentified (probably non-sporomorphous) elements.
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