Effects of two adipokinetic hormones (Pyrap-AKH and Peram-CAH-II) on the presence of diacylglycerol (DG) molecular species and their fatty acid (FA) constituents in the haemolymph of the firebug Pyrrhocoris apterus were investigated using liquid chromatography (HPLC) and electrospray ionization mass spectrometry (ESI-MS). The results show that DGs with characteristic FAs are preferentially mobilized from the fat body (FB) by the action of both the AKHs produced by P. apterus. Both the macropterous and brachypterous morphs have similar DG and FA profiles. A difference in the action of the Pyrap-AKH and the Peram-CAH-II, however, results in distinct differences in the distribution of FAs in the macropterous morph. It seems that C16 to a slight extent and unsaturated C18 FAs mainly play a dominant role in the AKH based action, in particular linoleic acid (18:2), which represents 50-60% of the total DG mobilized. The metabolically active C16 and C18 FAs are preferentially absorbed from the linden seeds and accumulated in the FB. The relationships between AKH action and FA distribution in DGs in P. apterus, compared to other insect species are summarized and discussed in detail.
Physico-chemical properties and carbohydrate-binding specificity of hemagglutination activity (HA) were compared in tissue lysates and haemolymph of unfed and bloodied females of five sandfly species. Sandfly gut lectins were found to be heat-labile, sensitive to dithiotreitol treatment, freezing/thawing procedures and were affected by divalent cations. The pH optimum of HA ranged between 7.0-7.5. Specificity of gut HA of all species studied was directed towards aminosugars and some glycoconjugates, mainly lipopolysaccharide from Escherichia coli K-235, heparin and fetuin. Gut HA of Phlebotomus papatasi (Scopoli, 1786) was strongly inhibited by lipophosphoglycan (LPG) from Leishmania major promastigotes. In females, that took blood, the HA was higher but the carbohydrate-binding specificity remained the same; this suggests that the same lectin molecule was present, at different levels, both in unfed and fed flies. High HA was found in ovaries of fed females of Lutzomyia longipalpis (Lutz et Nieva, 1912), P. papatasi and P. duhoscqi Neveu-Lemaire, 1906. In P. papatasi and P. duboscqi the HA was present also in the haemolymph and head lysates of both fed and unfed females. Carbohydrate-binding specificity of HA present in these tissues was similar with the gut lectin.
Reflex bleeding is one of the many anti-predation behavioural tactics used by insects. This behaviour is recorded widely in several insect taxa and provides scientists with an interesting opportunity for easily obtaining samples of haemolymph for physiological experiments. However, there is no good evidence that haemolymph and reflex blood have the same characteristics. In this study, we compared three basic characteristics of reflex blood and haemolymph collected from the body cavity, of three species of ladybirds: Harmonia axyridis, Coccinella septempunctata and Ceratomegilla undecimnotata. In the reflex blood we recorded the same concentration of haemocytes and total protein, and level of antimicrobial activity against Escherichia coli as in samples of haemolymph collected from within the body cavity. Therefore, reflex blood is a promising source of haemolymph for use in many physiological and immunological studies. It is the best way of collecting haemolymph if one wants to avoid challenging the immune system and concurrently also enables one to repeatedly collect haemolymph from one individual without injuring it, which would initiate repair mechanisms. The interspecific comparison indicates significant differences among the species studied in three characteristics measured. Interestingly, the native species Ceratomegilla undecimnotata has nearly as effective immune system as the invasive Harmonia axyridis based on the level of antibacterial activity against Escherichia coli.