Facultative diapause in the wax moth, Galleria mellonella, occurs in the final larval instar. Application of juvenile hormone analogs (JHAs) to the larvae of this species has similar effects to diapause, in terms of prolonged development of the larval stages and the arrest in the metamorphosis of internal organs. Here, we focus on testes development and spermatogenesis at the end of larval development in G. mellonella, how they are affected by diapause induced by an environmental decrease in temperature to 18°C and the application of a JHA (fenoxycarb) to larvae. Because neither testis development nor spermatogenesis are described in detail for this species, we examined them in individuals not in diapause during the period from the last larval instar to the newly emerged adult and present a timetable of changes that occur in the development of testes in this species. These observations have increased the very limited data on the course of spermatogenesis in pyralid insects. We then used these data for comparative analysis of testes in larvae from two experimental groups: individuals in diapause and those treated with fenoxycarb. The results on the general morphology testes revealed obvious degenerative changes caused by fenoxycarb (but not by diapause), including testicular wall hypertrophy and disarrangement of testicular follicles. Moreover, treatment with fenoxycarb finally resulted in the disintegration of nearly all testicular cyst-containing germ cells at different stages of spermatogenesis, a situation never previously described in the literature. In contrast, the main effect of diapause on testes was merely the degeneration of spermatocytes in the proximal regions of the testicular follicles. Finally, the TUNEL analyses, revealed that the degenerative changes in germ cells were apoptotic in character in the testes of both individuals in diapause and fenoxycarb-treated males., Piotr Bebas, Bronislaw Cymborowski, Michalina Kazek, Marta Anna Polanska., and Obsahuje bibliografii
The aim of this study was to investigate whether haemocytes of Galleria mellonella (Lepidoptera: Pyralidae) larvae produce reactive oxygen species (ROS) like human blood phagocytes. The production of ROS was measured first using luminol-enhanced chemiluminescence of un-stimulated and stimulated (four activators with different modes of action) haemolymph or isolated haemocytes. However, spontaneous and activated production of ROS remained at the background level. In subsequent experiments an ultrasensitive fluorescence method using Amplex Red reagent to detect hydrogen peroxide (H2O2) was used. After optimization, Amplex Red was successfully used for determining H2O2 production by both un-stimulated and stimulated haemocytes. To determine the affect of pH and ions on the measurement, several diluent solutions were tested. This revealed that Ca2+ and Mg2+ ions are less important for the reaction in insect than mammalian cells. Among the activators tested, phorbol myristate acetate (PMA) and calcium ionophore (Ca-I) had the best stimulatory effect on insect samples, while opsonised zymosan particles (OZP) was the best activator for human phagocytes. In conclusion, the haemocytes of G. mellonella produce H2O2 as an important innate immunity factor, but under different conditions and in different amounts, which probably results in them being less effective in killing microbes than human phagocytes. and Ondřej Vašíček, Ivana Papežíková, Pavel Hyršl.
1_The young larvae of insects living on dry food produce large amounts of water by the metabolic combustion of dietary lipids. The metabolic production of water needed for larval growth, previously known as hypermetabolic responses to juvenile hormone (JH), is associated with a 10- to 20-fold increase in the rate of O2 consumption (10,000 µl O2/g/h in contrast to the usual rate of 500 µl O2/g/h). Growing and moulting larvae are naturally hypermetabolic due to the endogenous release of JH from the corpora allata. At the last, larval-pupal or larval-adult moult there is no JH and as a consequence the metabolic rate is much lower and the dietary lipid is not metabolized to produce water but stored in the fat body. At this developmental stage, however, a hypermetabolic response can be induced by the exogenous treatment of the last larval instars with a synthetic JH analogue. In D. vulpinus, the JH-treated hypermetabolic larvae survive for several weeks without moulting or pupating. In T. castaneum and G. mellonella, the JH-treated hypermetabolic larvae moult several times but do not pupate. All these larvae consume dry food and the hypermetabolic response to JH is considered to be a secondary feature of a hormone, which is produced by some subordinated endocrine organ., 2_The organ is most probably the controversial prothoracic gland (PG), which is a typical larval endocrine gland that only functions when JH is present. According to our hypothesis, PG activated by JH (not by a hypothetical PTTH) releases an adipokinetic superhormone, which initiates the conversion of dietary lipid into metabolic water. This type of metabolic combustion of dietary lipid produces large quantities of endothermic energy, which is dissipated by the larvae in the form of heat. Thermovision imaging revealed that the body of hypermetabolic larvae of G. mellonella can be as hot as 43°C or more. In contrast, the temperature of "cold" normal last instar larvae did not differ significantly from that of their environment. It is highly likely that thermovision will facilitate the elucidation of the currently poorly understood hormonal mechanisms that initiate the production of metabolic water essential for the survival of insects that live in absolutely dry conditions., Karel Sláma, Jan Lukáš., and Obsahuje seznam literatury
Galleria mellonella larvae spin protective tubes, which they use until they finish feeding, when they spin cocoons. A feeding choice experiment showed that some of the silk produced by feeding larvae was consumed in addition to the standard diet (STD). To determine the effect of feeding on silk, last instar larvae were fed for 24 h on foods based on STD but modified by replacing the dry milk component (10% of the diet) with equal amounts of different kinds of silk. While each control larva consumed 21 ± 0.5 mg of the STD and produced 4.5 ± 0.1 mg of silk, larvae that ate the food that contained larval silk consumed 10 ± 0.4 mg of food and produced 6.1 ± 0.1 mg silk; the percentage ratio of silk produced to diet consumed was 21% and 61%, respectively. A more pronounced reduction in food consumption occurred when larvae were supplied with Galleria "cocoon" silk or the sericin fraction of such silk, and only 3.8 mg/larva was ingested of the diet containing Bombyx mori cocoon silk or its sericin fraction. Silk production expressed in terms of percentage of diet consumed was always higher than that recorded for larvae fed STD. We conclude that G. mellonella larvae recycle part of the silk that they produce during feeding. Presence of silk in the diet reduces food intake but increases the ratio of silk production to diet consumption. Sericin fraction of the cocoon silk seems to deter feeding., Haq Abdul Shaik, Archana Mishra, František Sehnal., and Obsahuje bibliografii