With the appearance of bovine spongiform encephalopathy (BSE) and a new variant of Creutzfeldt-Jakob disease (nvCJD) that seems to be caused by BSE, there is an increased need for improvement of diagnostic techniques and recognition of all variants of prion diseases in humans and animals. Publications on the immunohistochemical identification of PrP(Sc) in the tonsils and appendix in the incubation period of nvCJD indicate that new and more sensitive techniques for the detection of PrP(Sc) in various tissues may be a valuable tool for early diagnosis in prion diseases. We developed a new and sensitive technique to detect PrP(Sc) in formalin-fixed and paraffin-embedded tissue, the paraffin-embedded tissue blot (PET blot), and reinvestigated archival brain material from CJD as well as BSE and scrapie. In addition, C57/Bl6 mice experimentally infected with the ME7 strain were investigated sequentially during the incubation time to compare this new technique with conventional methodologies. The PET blot detects PrP(Sc) in idiopathic (sporadic) and acquired prion diseases, even in cases with equivocal or negative immunohistochemistry, and is more sensitive than the conventional Western blot and histoblot techniques. The PET blot makes possible the detection of PrP(Sc) during the incubation period long before the onset of clinical disease and in prion disease variants with very low levels of PrP(Sc). In mice experimentally infected with the ME7 strain, the PET blot detects PrP(Sc) in the brain 30 days after intracerebral inoculation-145 days before the onset of clinical signs. Its anatomical resolution is superior to that of the histoblot technique. It may therefore be of particular interest in biopsy diagnosis. Thus it complements other tissue-based techniques for the diagnosis of prion diseases in humans and animals.

Composition and diversity of aeroterrestrial phototrophic microbial communities are up to now poorly understood. Here, we present a comparative study addressing the composition of algal communities on sandstone substrata based upon the analysis of rRNA gene clone libraries from environmental samples and crude cultures. From a west-facing, shaded wall area of the mediaeval castle ruin Gleichen (Thuringia, Germany), sequences mainly related to the green algae Prasiococcus and Trebouxia (Trebouxiophyceae) were retrieved. A south-west-facing, sun-exposed wall area was mainly colonized by Apatococcus and a Phyllosiphon-related alga. Just a few species, in particular Stichococcus-related strains, were ubiquitous in both areas. Samples from a basement vault exposed to low irradiance exhibited Chlorophyceae like Chromochloris and Bracteacoccus. Thus, most green algae on the daylight-exposed walls were affiliated to Trebouxiophyceae, whereas Chlorophyceae were dominant in samples taken from the site kept under low irradiance. Accordingly, cyanobacterial communities were different: the sun-exposed area was dominated by Synechococcus-related organisms, while on the shaded wall area, cyanobacteria were almost absent. The filamentous Leptolyngbya dominated samples from the basement vault. Scanning electron microscopy revealed endolithic algal morphotypes (coccoid algae and diatoms) dominant in open pores between mineral particles. Here, the organisms may be also involved in biogenic weathering of stone.

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Symbiont-bearing and non-symbiotic marine bivalves were used as model organisms to establish biosignatures for the detection of distinctive symbioses in ancient bivalves. For this purpose, the isotopic composition of lipids (δ13C) and bulk organic shell matrix (δ13C, δ34S, δ15N) from shells of several thiotrophic, phototrophic, or non-symbiotic bivalves were compared (phototrophic: Fragum fragum, Fragum unedo, Tridacna maxima; thiotrophic: Codakia tigerina, Fimbria fimbriata, Anodontia sp.; non-symbiotic: Tapes dorsatus, Vasticardium vertebratum, Scutarcopagia sp.). ∆13C values of bulk organic shell matrices, most likely representing mainly original shell protein/chitin biomass, were depleted in thio- and phototrophic bivalves compared to non-symbiotic bivalves. As the bulk organic shell matrix also showed a major depletion of δ15N (down to -2.2 ‰) for thiotrophic bivalves, combined δ13C and δ15N values are useful to differentiate between thio-, phototrophic, and non-symbiotic lifestyles. However, the use of these isotopic signatures for the study of ancient bivalves is limited by the preservation of the bulk organic shell matrix in fossils. Substantial alteration was clearly shown by detailed microscopic analyses of fossil (late Pleistocene) T. maxima and Trachycardium lacunosum shell, demonstrating a severe loss of quantity and quality of bulk organic shell matrix with time. Likewise, the composition and δ13C-values of lipids from empty shells indicated that a large part of these compounds derived from prokaryotic decomposers. The use of lipids from ancient shells for the reconstruction of the bivalve's life style therefore appears to be restricted.

Carbonate minerals such as dolomite, kutnahorite or rhodochrosite are frequently, but not exclusively generated by microbial processes. In recent anoxic sediments, Mn(II)carbonate minerals (e.g. rhodochrosite, kutnahorite) derive mainly from the reduction of Mn(IV) compounds by anaerobic respiration. The formation of huge manganese-rich (carbonate) deposits requires effective manganese redox cycling in an oxygenated atmosphere. However, putative anaerobic pathways such as microbial nitrate-dependent manganese oxidation, anoxygenic photosynthesis and oxidation in ultraviolet light may facilitate manganese cycling even in an early Archean environment, without the availability of oxygen. In addition, manganese carbonates precipitate by microbially induced processes without change of the oxidation state, e.g. by pH shift. Hence, there are several ways how these minerals could have been formed biogenically and deposited in Precambrian sediments. We will summarize microbially induced manganese carbonate deposition in the presence and absence of atmospheric oxygen and we will make some considerations about the biogenic deposition of manganese carbonates in early Archean settings.

Re-colonization of freshly cleaned surfaces by aeroterrestrial microbial communities is up to now poorly understood. Here, we present a comparative study addressing the composition of algal and fungal communities on a marble sculpture, based upon the analysis of 18S rRNA gene clone libraries from environmental samples. The samples were taken from a blackish and greenish biofilm cover before surface cleaning and 1 year after cleaning treatment, when traces of re-colonization became visible to the naked eye. The composition of the fungal community indicated clear differences between the old grown biofilm and the treated surface. While the former was dominated by the ascomycetes Rhinocladiella, Glyphium and Capnodiales, the black yeast Sarcinomyces was clearly dominant 1 year after cleaning, but could not be retrieved from the old grown black biofilm. The green algal community was dominated by different phylotypes of the lichen algae Trebouxia, as well as the cosmopolitan green algae Apatococcus and Stichococcus. No essential differences in the green algal community before and after cleaning could be observed.

Siboglinid worms were sampled from four mud volcanoes in the Gulf of Cádiz (El Cid MV, Bonjardim MV, Al Gacel MV and Anastasya MV). These invertebrates are characteristic to cold seeps and are known to host chemosynthetic endosymbionts in a dedicated trophosome organ. However, little is known about their tube as a potential niche for other chemosynthetic and non-chemosynthetic microorganisms. Analyses by scanning and transmission electron microscopy showed dense biofilms on the tube in Al Gacel MV and Anastasya MV specimens by prokaryotic cells. Methanotrophic bacteria were the most abundant forming these biofilms as further confirmed by 16S rRNA sequence analysis. Furthermore, elemental analyses with electron microscopy and EDX point to the progressive mineralization of the biofilm and the tube in absence of nutrients. Environmental bacterial and archaeal 16S rRNA sequence libraries revealed abundant microorganisms related to these siboglinid worms and variation in microbial communities among samples. We argue that these differences must be related to variance in seepage activity, as it is the main source of nutrients. Thus, the tube remarkably increases the microbial biomass related to the worms and needs to be incorporated as an important part of the worm’s microbiota. Furthermore, empty tubes may still influence the composition of the active microbial community at those sites.

Molecular methods allow a comprehensive view on uncultured microbial communities in dimension stone. In the presented study, we focus on depth profiles of microbial colonization in sandstones with different porosity and overall durability. The sandstones were exposed to the environment for several years and were covered with a dense brown-greenish biofilm. Samples from the stone surface, from 0.5 cm and from 3.0 cm depths were taken under sterile conditions and subjected to analysis of microbial DNA (sequencing of 18S rRNA genes) and culturing experiments. A silica cemented Buntsandstein was just colonized at the stone surface, predominantly with algal and fungal microorganisms. Here, no environmental DNA could be isolated from depth profiles. From a calcitic Chirotheriensandstein with high pore size (shown by thin sections), environmental DNA from depths down to 3 cm could be retrieved. Apart from filamentous fungi and algae, mosses clearly dominated the microbial community. Hence, moss rhizoids or protonemata must be abundant as endoliths inside the stone material. It is reasonable to assume that the rhizoids may contribute to an increase in pore size by active penetration of the clastic material, even though colonization of the surface by mosses may not be necessarily obvious. This feature may imply stronger impact of stone decay induced by endolithic growth of bryophytes than hitherto observed.