A global decline in pollinator abundance and diversity has demanded increased research attention to the ecology and genetics of bumblebees. However, as progressively more restrictions are placed on sampling for insects, researchers are increasingly obliged to use archival specimens collected for purposes other than genetic analyses. In this study we assessed the suitability, for population genetic studies, of popular, low-cost methods for preservation and storage of bumblebee specimens. Specimens of Bombus terrestris L. were held under six storage regimes for up to two years. DNA was extracted from the samples using three extraction protocols and the quality of the DNA was examined using PCR amplification of a mitochondrial and a nuclear gene. All extraction and storage methods provided sufficient DNA for successful PCR amplification. However, samples preserved in acetone or at freezing temperatures yielded the highest DNA concentrations. DNA yields from pinned specimens at room temperature declined over time, particularly when using standard extraction techniques. DNA concentrations were significantly lower from specimens preserved in 70% ethanol compared to all other extraction techniques and declined linearly over the two years of storage. These results indicate that two of the most popular insect storage methods (pinning and storage in ethanol) should be avoided for the long-term preservation of genetic material for future studies. We suggest that optimal insect preservation methods should be incorporated into research protocols in order to best capitalise on limited collection opportunities., António S. Moreira ... []., and Obsahuje seznam literatury
Rhythmic body movements, cyclic gas exchange and heart activity were studied at low temperature (mostly at 5°C) in bumblebee Bombus terrestris foragers. Simultaneous measurements by means of an electrolytic respirometer combined with an optical system by infrared radiation, revealed a co-ordination between discontinuous gas exchange cycles and body rhythmic movements. No distinct correlation was found between intermittent heart activity and body movements. The carbon dioxide bursts were actively ventilated by abdominal vigorous pumping movements. These bursts followed each other with intervals of 25-30 minutes.
The periods of heart activity regularly alternated with heart pauses. The frequency of the heartbeat was similar to that of the weak abdominal pulsations (0.2-0.25 Hz) which were imperceptible with the naked eye. All bumblebees displayed abdominal pulsations which were independent of vigorous pumping movements. In bumblebee B. terrestris foragers haemolymph oscillation was assumed at 5°C, as is known in the honeybee and some other Bombus species.
Using an opto-cardiograph combined with an infrared gas analyzer regular bouts of abdominal contractions were recorded from last instar larva of Bombus terrestris. The rate of CO2 release was about 0.7 ml g-1 h-1. The bouts of contractions were of two types: weak extracardiac pulsations and vigorous pumping. The frequencies of pulsations and pumping were 25-35 per min and 8-12 per min, respectively. Bouts of extracardiac pulsations and abdominal pumping were independent of each other and sometimes overlapped. Cardiac contractions (heartbeats) were continuous (57-63 pulses/min). This study suggests that the periodically occurring abdominal contractions play an essential role in respiration and/or in haemolymph circulation in larvae of B. terrestris.
In many species sperm competition selects for large ejaculates while females eliminate or disable a high percentage of the spermatozoa in their storage organs in order to control paternity. Therefore, sperm viability is an excellent measure of the reproductive success of both males and females. Here we assess the viability of spermatozoa in males, freshly mated queens and old queens at the end of a colony cycle of the monandrous bumblebee Bombus terrestris. We found that the accessory testes of males contained a significantly lower percentage of dead sperm (i.e. higher average viability) than the spermathecae of both freshly mated and old queens. In each case, however, the percentage of dead spermatozoa was very small. No differences could be detected between sperm viability in freshly mated and old queens. To test for the possible incompatibility of the sperm and the environment provided by female spermathecae males and females either from different geographic regions (north and south of the Alps) or the same region were mated. We did not find any differences between matings of individuals from within or between regions. The mechanism that causes the lower sperm viability in females remains unknown.