Megabenthic assemblages at the southern Central Indian Ridge - Spatial segregation of inactive hydrothermal vents from active-, periphery- and non-vent sites.

Active hydrothermal vents are small-scale habitats hosting endemic fauna in a well-defined zonation around fluid effluents. The fauna of inactive hydrothermal vents and its relation to active vents and non-vent area is poorly known. Characterizing inactive areas is prerequisite to establish protected areas, especially in the context of potential seafloor massive sulfide mining, which targets inactive sites. Hierarchical clustering and Distance-based Redundancy Analysis revealed five assemblages, with significantly associated substrate types: I) active hydrothermal vent, II) periphery, III) inactive hydrothermal vent and IV) soft- and V) hard-substrate within the non-vent area. For the first time, a unique inactive faunal assemblage could be identified within the hydrothermally extinct inactive Gauss field and on adjacent hard substrates. The spatial separation from the active Edmond field and periphery and the non-vent area indicates the existence of an inactive assemblage.

A 3D imaging and visualization workflow, using confocal microscopy and advanced image processing for brachyuran crab larvae.

Confocal laser scanning microscopy is an excellent tool for nondestructive imaging of arthropods and can provide detailed information on morphology including fine surface detail. A methodology is presented here for the visualization by confocal microscopy of arthropods, using brachyuran crab zoeal stages as examples and postprocessing techniques derived from micro-CT protocols to improve the final images. This protocol is divided into description of the preprocessing steps (cleaning, staining, digesting and mounting), confocal laser scanning microscopy and data visualization using open-source, freeware programs ImageJ and Drishti. The advantages of using ImageJ to standardize stack data and Drishti for surface rendering are discussed. The methodology has been comprehensively tested using data acquired from all four brands of confocal microscope (Leica, Nikon, Olympus and Zeiss).