Investigation of the reproductive system in calanoid copepods: A new approach using 3D reconstructions from serial semi-thin cross-sections in Calanus glacialis and Metridia longa.

The structure of the female reproductive system of the calanoid copepods Calanus glacialis and Metridia longa from the White Sea was studied using light microscopy, scanning and transmission electron microscopy, as well as confocal laser scanning microscopy. For the first time, we applied also the method of 3D reconstructions from semi-thin cross-sections to visualize the general plan of the reproductive system in both species. The application of a combination of methods provided novel and detailed information on the genital structures and muscles located in the genital double-somite (GDS) as well as structures used for the reception and storage of spermatozoa, fertilization and release of eggs. An unpaired ventral apodeme and associated muscles located in the GDS are described for the first time for calanoid copepods. The role of this structure in copepod reproduction is discussed. Stages of oogenesis and the mechanism of yolk formation in M. longa are studied using semi-thin sections for the first time. A combination of non-invasive (LM, CLSM, SEM) and invasive techniques (semi-thin sections and TEM) applied in this study substantially improves our understanding of the functioning of the genital structures in calanoid copepods and could be recommended as a standard set of methods for future research in the reproductive biology of copepods.

Benchmarking DNA isolation methods for marine metagenomics.

Metagenomics is a powerful tool to study marine microbial communities. However, obtaining high-quality environmental DNA suitable for downstream sequencing applications is a challenging task. The quality and quantity of isolated DNA heavily depend on the choice of purification procedure and the type of sample. Selection of an appropriate DNA isolation method for a new type of material often entails a lengthy trial and error process. Further, each DNA purification approach introduces biases and thus affects the composition of the studied community. To account for these problems and biases, we systematically investigated efficiency of DNA purification from three types of samples (water, sea sediment, and digestive tract of a model invertebrate Magallana gigas) with eight commercially available DNA isolation kits. For each kit-sample combination we measured the quantity of purified DNA, extent of DNA fragmentation, the presence of PCR-inhibiting contaminants, admixture of eukaryotic DNA, alpha-diversity, and reproducibility of the resulting community composition based on 16S rRNA amplicons sequencing. Additionally, we determined a "kitome", e.g., a set of contaminating taxa inherent for each type of purification kit used. The resulting matrix of evaluated parameters allows one to select the best DNA purification procedure for a given type of sample.

Seasonal dynamics and life histories of three sympatric species of Pseudocalanus in two Svalbard fjords.

Small copepods are the most diverse and numerous group in high-latitude zooplankton, yet our knowledge of important species remains poor because of the difficulties involved in correct species identification. In this study, we use a molecular method of identification, a species-specific polymerase chain reaction, to provide the first description of the seasonal dynamics and life histories of the important genus Pseudocalanus in two Svalbard fjords with contrasting environments. We conducted monthly investigations in the relatively warm and ice-free Adventfjorden, supplemented with seasonal samples from the colder, seasonally ice-covered Billefjorden. We found three species of Pseudocalanus (the Arctic P. acuspes and P. minutus, and the boreal P. moultoni). Pseudocalanus acuspes had a distinct annual life cycle and dominated during summer, when it actively reproduced. Surprisingly, the boreal P. moultoni was present year-round in both fjords and was the dominant species during winter; the presence of all life stages of this species throughout the year suggests a more continuous reproduction. The Arctic P. minutus was the rarest of the three species and was likely able to complete its life cycle in Billefjorden but not in Adventfjorden. Our study demonstrates that closely related species may have different life strategies and environmental preferences, which presumably make high-latitude zooplankton communities more resilient to climate change impacts on genus but not necessarily on species level.