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When studying the vectorship of Culicoides species during the outbreak of Bluetongue disease (BTD) in Central Europe, the question arose whether the most common species and additionally proven vectors of BTV (C. obsoletus and C. pulicaris) are definitive species or do they belong to so-called complexes, since the determination based on morphological criteria is not very significant and knowledge on the life cycles is poor or even absent. Therefore, the present molecular biological study on their ITS-1, ITS-2 and 18SrDNA characteristics was initiated to investigate specimens, which had been determined by their wing morphology during an entomological monitoring in the years 2007 and 2008 at 91 farms in Germany (Mehlhorn et al. 2009). This study revealed novel types respectively different forms, which appeared very similar to Culicoides obsoletus, but showed slightly varying wing patterns. The molecular biological data were compared to those in data banks and combined to provisional dendrograms. The ITS-1 and ITS-2 analysis showed that the specimens determined in the monitoring as C. obsoletus inclusive those with different wing pattern correlate significantly with the data of C. obsoletus in the data banks and surrounded the data bank specifications of C. montanus and C. scoticus so closely that the latter might be only hardly separate species. A similar interpretation can also be drawn when looking at the 18S rDNA dendrogram. Thus, C. scoticus and C. montanus might be races of C. obsoletus rather than separate species. With respect to the ITS-1 and ITS-2 characteristics of C. pulicaris females, which morphologically and by size can be significantly differentiated from C. obsoletus, it was seen that this species is significantly situated on another rame of the dendrograms and in very close relationship to C. punctatus and C. lupicaris, so that the latter might also be only races of C. pulicaris. One of the two other most common species found in Northrhine-Westfalia-C. festivipennis-belongs to the rame of the dendrogram, where C. pulicaris is situated close to C. circumscriptus, while the other common species (C. nubeculosus) has its place close to C. puncticollis and C. variipennis on the rame, where C. obsoletus is found. Thus, this paper again clearly points out that the question "what is a definite species" is far from being solved, if the life cycle is not defined and morphology misleading. However, it also became clear that C. obsoletus and C. pulicaris are Europe-wide occurring species and that several other clearly described separate species are probably only races.

Besnoitia besnoiti tissue cysts from a recent outbreak in cattle in Germany were characterized with respect to their internal transcribed spacer regions 1, 2, and 18S rDNA gene sequences. These results were compared with own sequences of an Israelian isolate of B. besnoiti and of Besnoitia jellisoni cystozoites stored for years in liquid nitrogen. Furthermore, material was studied that was obtained from white mice (Balb/C) that had been successfully infected by intraperitoneal infection of fresh cystozoites from the German outbreak. All results were then compared and discussed with respect to databank sequences of other Besnoitia species. Comprehensive phylogenetic studies of B. besnoiti isolates from Germany revealed almost identical sequence alignments when compared to previously sequenced B. besnoiti isolates from Israel and Spain. More importantly, phylogenetic analysis revealed two distant clusters of Besnoitia species: the first one includes Besnoitia akodoni, Besnoitia darlingi, and Besnoitia oryctofelisi, while the second cluster includes B. besnoiti, Besnoitia bennetti, Besnoitia tarandi, and the Besnoitia species of rodents (B. jellisoni). The also B. jellisoni named species of the GenBank (AF 076860) must be another one, since our strain derives directly from Frenkel. These findings give strong hints that B. besnoiti has a cycle between rodents and a predator and that cattle and other are only accidental hosts.

A cDNA library was constructed from the glacier-dwelling eutardigrade Hypsibius klebelsbergi from more than 2000 individuals collected in the Austrian Central Alps. RNA, DNA and proteins were successively isolated by the Trizol®-method. From the RNA preparation a cDNA library was constructed with the cDNA inserted unidirectionally in the phagemid expression vector TriplEx2. The primary gene library had a titre of 107 pfu ml-1 and the final amplified gene library a titre of 6×109 pfu ml-1. The average insert length was about 1.6 kb. The partial sequence of H. klebelsbergi actin (746 bp) showed highest similarity to GenBank data of Drosophila melanogaster actin at the nucleic acid level (84.9%) and at the amino acid level (98%). Compared with actin fragments of the eutardigrades Ramazzottius oberhaeuseri (450 bp) and Macrobiotus sp. (453 bp) the identities were 85% - 81% and 100% - 98% with respect to the nucleic/amino acids. Identity with actin fragments (359 bp) of Hypsibius dujardini from GenBank was 96% - 100%

The actin gene of tardigrades was sequenced and analysed using a k ZAP Express cDNA library from the eutardigrade Hypsibius klebelsbergi previously constructed by us. We obtained the complete actin coding sequence of one isoform (1128 bp; 375 amino acids; MW 41 674 Da) together with parts of the 3¢-and 5¢-UTR region. Comparison of the 12 incomplete actin sequences of Hypsibius dujardini incorporated in GenBank indicates that this H. klebelsbergi actin sequence probably represents the most abundant muscle isoform. Ten of the H. dujardini clones show minor differences in codon usage and identical amino acid compositions to the H. klebelsbergi actin. Only two clones show amino acid variations in one and five positions, respectively, but show identical amino acids at their N-terminus. A considerable similarity between the 5¢-and 3¢-UTR regions of both tardigrade species was recognized. The H. klebelsbergi actin exhibits an overall high sequence similarity to the vertebrate b-actin. A comparison of muscle actins from various vertebrates as well as Ecdysozoa and non-Ecdysozoa revealed a more pronounced similarity of the tardigrade actin to arthropods and annelids and not to nematodes.