Biogeography of limno-terrestrial Tardigrada, with particular reference to the Antarctic fauna

The Tardigrada is a cosmopolitan phylum of pre-Pangaean origin, yet tardigrade families and genera show distinct biogeographic components isolated by two major geological events. Separate Laurasian and Gondwanan familial clusters correlate with the Triassic disintegration of Pangaea, while discrete Antarctic, Australian and New Zealand familial/generic clusters relate to the subsequent Jurassic/Cretaceous disintegration of Gondwana.     

Milnesium berladnicorum sp. n. (Eutardigrada,Apochela, Milnesiidae), a new species of water bear from Romania

In a lichen sample collected from a tree in Bârlad town (Vaslui County, Romania), a new tardigrade species belonging to the genus Milnesium (granulatum group) was found. Milnesium berladnicorum sp. n. is most similar (in the type of dorsal sculpture) to Milnesium beasleyi Kaczmarek et al., 2012 but differs from it mainly by having a different claw configuration and some morphometric characters. Additionally, the new species differs from other congeners of the granulatum group by the different type of dorsal sculpture, claw configuration and some morphometric characters.     

Autophagy as the cell survival in response to a microsporidian infection of the midgut epithelium of Isohypsibius granulifer granulifer (Eutardigrada: Hypsibiidae)

The midgut epithelial cells of many invertebrates may possess microorganisms which act as symbionts or pathogens (bacteria, microsporidia, viruses). During our previous studies on Isohypsibius granulifer granulifer Thulin, 1928 (Tardigrada, Eutardigrada), which examined alterations of the midgut epithelium during oogenesis, we found that some of the specimens were infected with microsporidia. All stages of pathogens occurred in the cytoplasm of the digestive cells in the midgut epithelium of I. g. granulifer that were infected with microsporidia: meronts, sporonts, sporoblasts, and spores. The cytoplasm of the digestive cells was rich in mitochondria, cisterns of rough endoplasmic reticulum (RER), and Golgi complexes. Autophagy in the digestive cells of the dorsal midgut was much more intensive in comparison with noninfected specimens. Membranes of phagophores surrounded the pathogens forming autophagosomes. These latter structures fused with lysosomes forming autolysosomes and residual bodies appeared. Neither glycogen granules nor droplets of varying electron density, which accumulated in digestive cells during vitellogenesis and choriogenesis, appeared in individuals with microsporidia. While the midgut epithelium in noninfected specimens takes part in vitellogenesis and choriogenesis, in infected specimens, midgut cells are involved in the process of autophagy as a survival strategy.     

New molecular data for tardigrade phylogeny, with the erection of Paramacrobiotus gen. nov.

Up to few years ago, the phylogenies of tardigrade taxa have been investigated using morphological data, but relationships within and between many taxa are still unresolved. Our aim has been to verify those relationships adding molecular analysis to morphological analysis, using nearly complete 18S ribosomal DNA gene sequences (five new) of 19 species, as well as cytochrome oxidase subunit 1 (COI) mitochondrial DNA gene sequences (15 new) from 20 species, from a total of seven families. The 18S rDNA tree was calculated by minimum evolution, maximum parsimony (MP) and maximum likelihood (ML) analyses. DNA sequences coding for COI were translated to amino acid sequences and a tree was also calculated by neighbour-joining, MP and ML analyses. For both trees (18S rDNA and COI) posterior probabilities were calculated by MrBayes. Prominent findings are as follows: the molecular data on Echiniscidae (Heterotardigrada) are in line with the phylogenetic relationships identifiable by morphological analysis. Among Eutardigrada, orders Apochela and Parachela are confirmed as sister groups. Ramazzottius (Hypsibiidae) results more related to Macrobiotidae than to the genera here considered of Hypsibiidae. Macrobiotidae and Macrobiotus result not monophyletic and confirm morphological data on the presence of at least two large groups within Macrobiotus. Using 18S rDNA and COI mtDNA genes, a new phylogenetic line has been identified within Macrobiotus , corresponding to the ' richtersi-areolatus group'. Moreover, cryptic species have been identified within the Macrobiotus ' richtersi group' and within Richtersius . Some evolutionary lines of tardigrades are confirmed, but others suggest taxonomic revision. In particular, the new genus Paramacrobiotus gen. n. has been identified, corresponding to the phylogenetic line represented by the ' richtersi-areolatus group'.     

Dynamics of Long-term Anhydrobiotic Survival of Lichen-dwelling Tardigrades

It is not rare to find in references that anhydrobiotic tardigrades can survive for more than a century. However, a closer look at the empirical evidence provides very little support that tardigrades are capable of surviving dried for such a long time. In order to resolve this discrepancy, we carried out a study to evaluate the long-term survival of naturally dried tardigrades. A large fragment of dry lichen (Xanthoria parietina) was collected in the field two days after a rainy day in 1999. The dry lichen was stored inside a paper bag in the laboratory at room temperature and humidity and under atmospheric oxygen exposure. Replicates of the dry lichen were re-hydrated after various time periods of storage, with all tardigrades extracted and the survivors enumerated. Five species of tardigrades were found, but two of them only occasionally. Ramazzottius oberhaeuseri, Echiniscus testudo and Echiniscus trisetosus were sufficiently represented for statistical analysis. At the beginning of the experiment 91.1% of R. oberhaeuseri and 71.7% of Echiniscus spp. were alive. R. oberhaeuseri survived up to 1604 days, while Echiniscus spp. lived up to 1085 days. Recovery after four years of anhydrobiosis has to be considered a very good long-term survival, which is important from an ecological and evolutionary point of view.     

Die Struktur des Nervengewebes von Macrobiotus hufelandi C.A.S. Schultze (Tardigrada)

Zusammenfassung Das Nervengewebe von Macrobiotus hufelandi zeichnet sich durch stark verzweigte pseudunipolare Nervenzellen und relativ wenige Gliazellen aus. Die Neurone besitzen rauhes ER, freie Ribosomen, zahlreiche Mitochondrien, einen wenig ausgeprägten Golgi-Apparat und einen elektronenlichten Kern. In ihren Axonen finden sich Vesikel und Einschlüsse unterschiedlicher Größe und Zusammensetzung. Die Gliazellen verzweigen sich stark und besitzen glattes ER, viele freie Ribosomen und einen elektronendichten Kern. Ganglien und Nerven werden nur durch eine dünne Neurallamelle vom extraganglionären Raum getrennt. Die morphologische Ausbildung des Nervengewebes wird im Hinblick auf die extreme Lebensweise der Tardigraden diskutiert.

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The structure of the nervous tissue in Macrobiotus hufelandi C. A. S. schultze (tardigrada)

Summary The typical elements of the nervous tissue of Macrobiotus hufelandi are strongly ramified pseudunipolar neurons and a small amount of glial cells. In the perikarya of neurons there are rough ER, free ribosomes, many mitochondria, a poor Golgi-apparatus, and an electron-light nucleus. Nerve fibers contain masses of vesicles and inclusions of different size and composition. The ramifying glial cells have smooth ER, many free ribosomes, and an electron-dense nucleus. Ganglia and nerves are separated from the extraganglionic cavity by a thin acellular sheath (neural lamella). The organization of the nervous tissue is discussed with regard to the extreme conditions of environment of the tardigrades.

     

Echiniscus testudo (Doyère, 1840) in New Zealand: anthropogenic dispersal or evidence for the ‘Everything is Everywhere’ hypothesis?

Many species of microscopic invertebrates, such as tardigrades or rotifers, have been traditionally regarded as cosmopolitan. This conviction was based on classical taxonomic observations of very similar phenotypes across the globe and hypothetical abilities of micrometazoans for large-scale dispersal with air and water currents. However, several recent studies showed that micrometazoan species may have restricted geographic ranges, suggesting that microscopic size is not the only determinant of dispersal. Here, we describe the first Australasian record of Echiniscus testudo, a heterotardigrade originally described from the Holarctic. Morphological and genetic analyses gave congruent conclusions and confirmed that European and New Zealand populations represent a single species. Importantly, however, without broader sampling in primeval localities, it is not possible to test whether the New Zealand record of E. testudo is a result of natural dispersal or whether the species was brought to Australasia by humans.     

缓步动物门(Tardigrada)研究进展

本文简要概述了20世纪以来缓步动物在分类学、生物学和生态学的研究进展和成就.按照缓步动物前沿研究的发展趋势,结合我国的研究现状,作者提出了几点建议.     

Identification and differentiation of Heterotardigrada and Eutardigrada species by riboprinting

In the last decades, the number of known tardigrade species has considerably increased to more than 960 species with new ones being discovered every year. However, the study of tardigrade species presents a general problem which is frequently encountered during the work with invertebrates: small size and remarkable degrees of phenotypic plasticity may sometimes not permit a definite identification of the species. In this investigation we have used riboprinting, a tool to study rDNA sequence variation, in order to distinguish tardigrade species from each other. The method combines a restriction site variation approach of ribotyping with amplified DNAs. In eight investigated species of heterotardigrades and eutardigrades we have amplified the genes for the small subunit ribosomal RNA (SSU; 18S) and subsequently sequenced the genes. Virtual riboprints were used for identification of restriction sites from ten already published 18S rDNA sequences and seven new 18S rDNA sequences. On the basis of the obtained sequences, diagnostic restriction fragment patterns can be predicted with only 11 restriction enzymes. The virtual digestion confirmed the obtained restriction fragment patterns and restriction sites of all amplified and digested tardigrade DNAs. We show that the variation in positions and number of restriction sites obtained by standard restriction fragment analysis on agarose gels can be used successfully for taxonomic identification at different taxonomic levels. The simple restriction fragment analysis provides a fast and convenient method of molecular barcoding for species identification in tardigrades.