In solfataric fields in southwestern Iceland, neutral and sulfide-rich hot springs are characterized by thick bacterial mats at 60 to 80°C that are white or yellow from precipitated sulfur (sulfur mats). In low-sulfide hot springs in the same area, grey or pink streamers are formed at 80 to 90°C, and a Chloroflexus mat is formed at 65 to 70°C. We have studied the microbial diversity of one sulfur mat (high-sulfide) hot spring and one Chloroflexus mat (low-sulfide) hot spring by cloning and sequencing of small-subunit rRNA genes obtained by PCR amplification from mat DNA. Using 98% sequence identity as a cutoff value, a total of 14 bacterial operational taxonomic units (OTUs) and 5 archaeal OTUs were detected in the sulfur mat; 18 bacterial OTUs were detected in the Chloroflexus mat. Although representatives of novel divisions were found, the majority of the sequences were >95% related to currently known sequences. The molecular diversity analysis showed that Chloroflexus was the dominant mat organism in the low-sulfide spring (1 mg liter ؊1 ) below 70°C, whereas Aquificales were dominant in the high-sulfide spring (12 mg liter ؊1 ) at the same temperature. Comparison of the present data to published data indicated that there is a relationship between mat type and composition of Aquificales on the one hand and temperature and sulfide concentration on the other hand.Sulfide-rich hot springs with neutral or alkaline pH are relatively rare in most geothermal areas in the world. However, these types of hot springs are rather common in Iceland due to high ground water level and climatic conditions, i.e., from melting snow and rain. Bacteria that thrive in such springs often form long streamers or mats, but the appearance of the mats and the types of bacteria in them seem to vary depending on the sulfide concentration, pH, temperature, and other chemical and physical factors (6,7,10,11,12,20,33). Many Icelandic hot springs have sulfide concentrations as high as 30 mg liter Ϫ1 and, under such conditions, thick bacterial mats which can be spectacularly white or bright yellow from precipitated sulfur are formed.The diversity of many microbial ecosystems has now been studied with different molecular methods, such as analysis of small-subunit (SSU) rRNA by sequencing, denaturing gradient gel electrophoresis, or restriction fragment length polymorphism analyses. These studies show that the diversity of microbial ecosystems is typically 100 to 1,000 times greater than that shown by cultivation alone (14,15,22,23,30,31). The sequencing of rRNA genes from environmental samples is very informative, since it provides information on both the phylogenetic relationship and the population structure of the microbial community. With increased understanding of the role and importance of microbes in many ecosystems, the benefit of microbial diversity studies is being recognized. The practical value of these methods is already widespread, as they can be used to study the performance of wastewater treatment plants (27), monitor changes upo...

BackgroundInternational fish trade reached an import value of 62.8 billion Euro in 2006, of which 44.6% are covered by the European Union. Species identification is a key problem throughout the life cycle of fishes: from eggs and larvae to adults in fisheries research and control, as well as processed fish products in consumer protection.Methodology/Principal FindingsThis study aims to evaluate the applicability of the three mitochondrial genes 16S rRNA (16S), cytochrome b (cyt b), and cytochrome oxidase subunit I (COI) for the identification of 50 European marine fish species by combining techniques of “DNA barcoding” and microarrays. In a DNA barcoding approach, neighbour Joining (NJ) phylogenetic trees of 369 16S, 212 cyt b, and 447 COI sequences indicated that cyt b and COI are suitable for unambiguous identification, whereas 16S failed to discriminate closely related flatfish and gurnard species. In course of probe design for DNA microarray development, each of the markers yielded a high number of potentially species-specific probes in silico, although many of them were rejected based on microarray hybridisation experiments. None of the markers provided probes to discriminate the sibling flatfish and gurnard species. However, since 16S-probes were less negatively influenced by the “position of label” effect and showed the lowest rejection rate and the highest mean signal intensity, 16S is more suitable for DNA microarray probe design than cty b and COI. The large portion of rejected COI-probes after hybridisation experiments (>90%) renders the DNA barcoding marker as rather unsuitable for this high-throughput technology.Conclusions/SignificanceBased on these data, a DNA microarray containing 64 functional oligonucleotide probes for the identification of 30 out of the 50 fish species investigated was developed. It represents the next step towards an automated and easy-to-handle method to identify fish, ichthyoplankton, and fish products.

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