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Acid mine drainage sites are extreme environments with high acidity and metal ion concentrations. Under anoxic conditions, microbial sulfate reduction may trigger the formation of secondary minerals as a result of H2S production and pH increase. This process was studied in batch experiments with enrichment cultures from acidic sediments of a pit lake using growth media set at different pH values and containing elevated concentrations of Fe²⁺ and Al³⁺. At initial pH values of 5 and 6, sulfate reduction occurred shortly after inoculation. Sulfate- reducing bacteria affiliated to the genus Desulfosporosinus predominated the microbial communities as shown by 16S rRNA gene analysis performed at the end of the incubation. At initial pH values of 3 and 4, sulfate reduction and cell growth occurred only after an extended lag phase, however, at a higher rate than in the less acidic assays. At the end of the growth phase, enrichments were dominated by Thermodesulfobium spp. suggesting that these sulfate reducers were better adapted to acidic conditions. Iron sulfides in the bulk phase were common in all assays, but specific aluminum precipitates formed in close association with cell surfaces and may function as a detoxification mechanism of dissolved Al species at low pH.

A total of 120 mycoplasma strains were recovered from 97 of 265 diseased seals investigated during the seal epidemic in the North Sea and in the Baltic Sea in 1988. Mycoplasmas were isolated from the respiratory tracts (including lungs), hearts, brains, and eyes of the seals. Thirty strains were filter cloned and investigated for their morphological, biochemical, and serological characteristics compared with the characteristics of previously described species. The results of an indirect immunofluorescence test, a growth inhibition test, and an immunobinding assay showed that these strains belong to two new species, for which the names MycopZasrna phocarhinis and Mycoplasrna phocacerebrde are proposed. M. phocarhinis (17 strains) did not ferment glucose or hydrolyze arginine but did reduce tetrazolium chloride and potassium tellurite and produced films and spots. M. phocacerebrale (13 strains) metabolized arginine but not glucose and produced phosphatase but did not reduce tetrazolium chloride and potassium tellurite. Both species lysed sheep erythrocytes but did not absorb sheep or guinea pig erythrocytes. The type strain of M. phocarhinis is strain 852 (= ATCC 49639), and the type strain of M. phocacerebrate is strain 1049 (= ATCC 49640).In 1988 mass mortality spread among the harbor seals (Phoca vitulina L.) of the Baltic Sea and the North Sea.More than 18,000 seals died because of an acute disease characterized by pneumonia, skin lesions, diarrhoea, polyarthritis, nervous signs, and abortions in pregnant females (9,10,12,16,17,(19)(20)(21). In addition to several viruses (9,10, 19-21), mycoplasmas were isolated from a large number of the diseased seals (12). Altogether 122 mycoplasma strains were recovered from 265 diseased seals investigated.In this paper we report the isolation details of these strains, describe our taxonomic examination of 30 selected strains compared with previously described species of Mollicutes, and discuss our findings in relation to the requirements for description of new species. MATERIALS AND METHODS Seals.Investigations for mycoplasmas were performed on 265 seals found moribund or dead along the coasts of Schleswig-Holstein and Lower Saxony (10,12,19). Organ materials from seals found dead were taken by M. Stede (Staatliches Veterinaruntersuchungsamt, Cuxhaven, Federal Republic of Germany). Samples from living animals were obtained from the Institute for Pathology, Tierarztliche Hochschule, Hannover, Federal Republic of Germany, where moribund seals were sacrificed for histological investigations, or from the seal orphanage at Norddeich, Federal Republic of Germany, where diseased seals were kept for treatment. Swabs were taken from noses, eyes, and throats of living animals. Samples of tracheae, lungs, hearts, livers, brains, kidneys, and spleens were obtained from dead or sacrificed animals.Cultivation of mycoplasmas. Homogenized organ materials and swabs were suspended in 2 ml of modified Friis medium (13). Portions (0.2 ml) of these suspensions were transferred to 2-ml portion...

Microbial Fe(III) and sulfate reduction are important electron transport processes in acidic pit lakes and stimulation by the addition of organic substrates is a strategy to remove acidity, iron and sulfate. This principle was applied in a pilot‐scale enclosure in pit lake 111 (Brandenburg, Germany). Because seasonal and spatial variation of temperature may affect the performance of in situ experiments considerably, the influence of temperature on Fe(III) and sulfate reduction was investigated in surface sediments from the enclosure in the range of 4–28 °C. Potential Fe(III) reduction and sulfate reduction rates increased exponentially with temperature, and the effect was quantified in terms of the apparent activation energy Ea measuring 42–46 kJ mol−1 and 52 kJ mol−1, respectively. Relatively high respiration rates at 4 °C and relatively low Q10 values (∼2) indicated that microbial communities were well adapted to low temperatures. In order to evaluate the effect of temperature on growth and enrichment of iron and sulfate‐reducing bacterial populations, MPN (Most Probable Number) dilution series were performed in media selecting for the different bacterial groups. While the temperature response of specific growth rates of acidophilic iron reducers showed mesophilic characteristics, the relatively high specific growth rates of sulfate reducers at the lowest incubation temperature indicated the presence of moderate psychrophilic bacteria. In contrast, the low cell numbers and low specific growth rates of neutrophilic iron reducers obtained in dilution cultures suggest that these populations play a less significant role in Fe and S cycling in these sediments. SSCP (Single‐Strand Conformation Polymorphism) or DGGE (Denaturing Gradient Gel Electrophoresis) fingerprinting based on 16S rRNA genes of Bacteria indicated different bacterial populations in the MPN dilution series exhibiting different temperature ranges for growth.

Iron- and sulfate-reducing microorganisms play an important role for alkalinity-generating processes in mining lakes with low pH. In the acidic mining lake 111 in Lusatia, Germany, a passive in situ remediation method was tested in a large scale experiment, in which microbial iron and sulfate reduction are stimulated by addition of Carbokalk (a mixture of the nonsugar compounds of sugar beets and lime) and straw. The treated surface sediment consisted of three layers of different pH and geochemical composition. The top layer was acidic and rich in Fe(III), the second and third layer both showed moderately acidic to circum-neutral pH values, but only the second was rich in organics, strongly reduced and sulfidic. Aim of the study was to elucidate the relative importance of neutrophilic heterotrophic, acidophilic heterotrophic, and acidophilic autotrophic iron-reducing microorganisms in each of the three layers. In order to distinguish between them, the effect of their respective characteristic electron donors acetate, glucose, and elemental sulfur on potential iron reduction rates was investigated. Limitation of iron reduction by the availability of Fe(III) was revealed by the addition of Fe(OH)(3). The three groups of iron-reducing microorganisms were quantified by most probable number (MPN) technique and their community composition was analyzed by cloning and sequencing of 16S rRNA genes. In the acidic surface layer, none of the three electron donors stimulated iron reduction; acetate even had an inhibiting effect. In agreement with this, no decrease of the added electron donors was observed. Iron reduction rates were low in comparison to the other layers. Iron reduction in layers 2 and 3 was enhanced by glucose and acetate, accompanied by a decrease of these electron donors. Addition of elemental sulfur did not enhance iron reduction in either layer. Layer 2 exhibited the highest iron reduction rate (4.08 mmol dm(-3) d(-1)) and the highest cell numbers in MPN media. In MPN enrichments from all layers, Acidithiobacillus-like sequences were frequent. In addition to these, sequences related to Fulvimonas and Clostridium dominated in layer 1. MPN enrichments of layer 2 were diverse, containing Rhodocyclaceae-related sequences and surprisingly low numbers of Geobacteraceae. In layer 3, Sulfobacillus and Trichococcus spp. were also important. It was concluded that in the surface layer mainly acidophilic, probably autotrophic and heterotrophic, iron reducers were active, whereas in layers 2 and 3 mainly neutrophilic heterotrophs were important for iron reduction. These differ from well-studied Fe(III) reducers in other environments, so they deserve further study. The potential for acid-producing sulfur-driven Fe(III) reduction seemed not to be critical for in situ remediation.