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
Diapausing larvae of Aphidoletes aphidimyza (Diptera: Cecidomyiidae) had relatively low supercooling points (SCP) ranging from -19.0 to -26.4°C. None of the specimens that froze at this temperature survived. A high survival rate (up to 87%) at -10°C for 10 days was observed in supercooled larvae. Such features are characteristic for insects that use a chill-tolerance strategy of cold hardiness. However, the cocoons formed by the diapausing larvae were penetrable by external ice crystals and the larvae showed a relatively high survival rate (23 - 34%) at -10°C for 10 days also in the frozen state caused by inoculation by external ice at high subzero temperatures. Such a duality with respect to cold hardiness strategies seems to be ecologically relevant to overwintering in soil habitats where there may be unpredictable contact with external ice.
The cocoons characteristic of the prepupal and pupal stages of many insects vary widely in size, durability, structure, shape and colour, as well as in other features such as orientation and attachment to the substrate. In some species they vary seasonally. Most cocoons provide little direct insulation, although they may reduce the rate at which temperature changes, but many provide the mechanical protection required for overwintering beneath insulating substrates such as soil and snow. The cocoons of some terrestrial species prevent inoculative freezing by isolating the integument from ice crystals on the cocoon surface or its surroundings. In some aquatic species, cocoons appear to limit damage by providing mechanical protection during the freezing of surrounding water. Some cocoons help in the acquisition of solar heat: dark structures are especially effective because dark pigments absorb heat, and surrounding layers trap this heat. Insects are immobilized when it is cold and so cannot move in response to environmental threats, and protective cocoons made for winter tend to be more robust than their summer counterparts. Such cocoons protect against abrasion of the waterproof layer of the cuticle. In some species, robust cocoons or complex structures impede natural enemies. Cocoon silk has anti-bacterial and anti-fungal actions. Other cocoons are more or less waterproof. These and other features withstand simultaneous constraints in addition to cold. Therefore, cocoons enhance survival during cold conditions in many species. However, this conclusion is based on fragmentary evidence, and there has been relatively little explicit examination of the roles of cocoons during winter. Therefore, specific work is required to assess resistance to or enhancement of inoculative freezing, resistance to penetration by natural enemies and water, the roles of particular cocoon silks and silk constituents, and the quantitative contributions of cocoons to winter survival in nature.