Selective elimination of bacterial faecal indicators in the Schmutzdecke of slow sand filtration columns.

Slow sand filtration (SSF) is an effective low-tech water treatment method for pathogen and particle removal. Yet despite its application for centuries, it has been uncertain to which extent pathogenic microbes are removed by mechanical filtration or due to ecological interactions such as grazing and competition for nutrients. In this study, we quantified the removal of bacterial faecal indicators, Escherichia coli and Enterococcus faecalis, from secondary effluent of a wastewater treatment plant and analysed the microbial community composition in compartments of laboratory model SSF columns. The columns were packed with different sand grain sizes and eliminated 1.6-2.3 log units of faecal indicators, which translated into effluents of bathing water quality according to the EU directive (<500 colony forming units of E. coli per 100 ml) for columns with small grain size. Most of that removal occurred in the upper filter area, the Schmutzdecke. Within that same zone, total bacterial numbers increased however, thus suggesting a specific elimination of the faecal indicators. The analysis of the microbial communities also revealed that some taxa were removed more from the wastewater than others. These results accentuate the contribution of biological mechanisms to water purification in SSF.

"Devil's gardens" in the Ecuadorian Amazon--association of the allelopathic tree Duroia hirsuta (Rubiaceae) and its "gentle" ants.

The relationship between the myrmecophytic tree Duroia hirsuta (Rubiaceae) and its associated, minute, non-aggressive Myrmelachista sp. (Formicinae) ants is poorly investigated, even though the small understory tree is surrounded by a conspicuos zone almost devoid of vegetation. As to the cause of these barely vegetated "Devil's Gardens", two alternative interpretations exist in the literature: the pruning activity of ants and allelopathy. We investigated the mutualistic Duroia-Myrmelachista system in the Ecuadorian Amazon, focusing on the phenomenon of the areas of reduced vegetation, as well as on the natural history of the associated ants. We conducted planting experiments and behavioral studies, including coccoids, the third partner in this mutualism. Because the well-studied Triplaris americana (Polygonaceae) possesses analogous vegetation free surroundings created by the assiduous mechanical pruning of its mutualistic ant partner (Pseudomyrmex sp.), parallel comparative investigations of this ant-plant association were conducted. The two systems are not equivalent; it was demonstrated that the remarkably inactive Myrmelachista ants are not responsible for the bare areas around Duroia plants. Instead, we regard allelopathy as the causative agent for the clear zones.

Heterotrophic symbionts of phototrophic consortia: members of a novel diverse cluster of Betaproteobacteria characterized by a tandem rrn operon structure.

Phototrophic consortia represent the most highly developed type of interspecific association of bacteria and consist of green sulfur bacterial epibionts attached around a central colourless rod-shaped bacterium. Based on 16S rRNA gene sequencing, the central bacterium of the consortium 'Chlorochromatium aggregatum' was recently shown to represent a novel and phylogenetically isolated lineage of the Comamonadaceae within the beta-subgroup of the Proteobacteria. To date, 19 types of phototrophic consortia are distinguished based on the different 16S rRNA gene sequences of their epibionts, but the diversity and phylogenetic relationships of the heterotrophic partner bacteria are still unknown. We developed an approach based on the specific rrn (ribosomal RNA) operon structure of the central bacterium of 'C. aggregatum' to recover 16S rRNA gene sequences of other central bacteria and their close relatives from natural consortia populations. Genomic DNA of the central bacterium of 'C. aggregatum' was first enriched several hundred-fold by employing a selective method for growth of consortia in a monolayer biofilm followed by a purification of the genome of the central bacterium by cesium chloride-bisbenzimidazole equilibrium density gradient centrifugation. A combination of inverse PCR, cloning and sequencing revealed that two rrn operons of the central bacterium are arranged in a tandem fashion and are separated by an unusually short intergenic region of 195 base pairs. This rare gene order was exploited to screen various natural microbial communities by PCR. We discovered a diverse and previously unknown subgroup of Betaproteobacteria in the chemoclines of freshwater lakes. This group was absent in other freshwater and soil samples. All the 16S rRNA gene sequences recovered are related to that of the central bacterium of 'C. aggregatum'. Fluorescence in situ hybridization indicated that two of these sequences originated from central bacteria of different phototrophic consortia, which, however, were only distantly related to the central bacterium of 'C. aggregatum'. Based on a detailed phylogenetic analysis, these central bacterial symbionts of phototrophic consortia have a polyphyletic origin.

Chlorobium chlorochromatii sp. nov., a symbiotic green sulfur bacterium isolated from the phototrophic consortium "Chlorochromatium aggregatum".

A symbiotic green sulfur bacterium, strain CaD, was isolated from an enrichment culture of the phototrophic consortium "Chlorochromatium aggregatum". The capability of the epibiont to grow in pure culture indicates that it is not obligately symbiotic. Cells are Gram-negative, nonmotile, rod-shaped and contain chlorosomes. Strain CaD is obligately anaerobic and photolithoautotrophic, using sulfide as electron donor. Acetate and peptone are photoassimilated in the presence of sulfide and hydrogencarbonate. Photosynthetic pigments contain bacteriochlorophylls a and c, and gamma-carotene and OH-gamma-carotene glucoside laurate as the dominant carotenoids. In cells from pure cultures, chlorosomes are equally distributed along the inner face of the cytoplasmic membrane. In contrast, the distribution of the chlorosomes in symbiotic epibiont cells is uneven, with chlorosomes being entirely absent at the site of attachment to the central bacterium. The symbiotic epibiont cells display a conspicuous additional layered structure at the attachment site. The G + C content of genomic DNA of strain CaD is 46.7 mol%. On the basis of 16S rRNA sequence comparison, the strain is distantly related to Chlorobium species within the green sulfur bacteria phylum (

Molecular characterization of the nonphotosynthetic partner bacterium in the consortium "Chlorochromatium aggregatum".

Phototrophic consortia represent valuable model systems for the study of signal transduction and coevolution between different bacteria. The phototrophic consortium "Chlorochromatium aggregatum" consists of a colorless central rod-shaped bacterium surrounded by about 20 green-pigmented epibionts. Although the epibiont was identified as a member of the green sulfur bacteria, and recently isolated and characterized in pure culture, the central colorless bacterium has been identified as a member of the beta-Proteobacteria but so far could not be characterized further. In the present study, "C. aggregatum" was enriched chemotactically, and the 16S rRNA gene sequence of the central bacterium was elucidated. Based on the sequence information, fluorescence in situ hybridization probes targeting four different regions of the 16S rRNA were designed and shown to hybridize exclusively to cells of the central bacterium. Phylogenetic analyses of the 1,437-bp-long sequence revealed that the central bacterium of "C. aggregatum" represents a so far isolated phylogenetic lineage related to Rhodoferax spp., Polaromonas vacuolata, and Variovorax paradoxus within the family Comamonadaceae. The majority of relatives of this lineage are not yet cultured and were found in low-temperature aquatic environments or aquatic environments containing xenobiotica or hydrocarbons. In CsCl-bisbenzimidazole equilibrium density gradients, genomic DNA of the central bacterium of "Chlorochromatium aggregatum" formed a distinct band which could be detected by quantitative PCR using specific primers. Using this method, the G+C content of the central bacterium was determined to be 55.6 mol%.

"Devil's gardens" in the Ecuadorian Amazon--association of the allelopathic tree Duroia hirsuta (Rubiaceae) and its "gentle" ants

The relationship between the myrmecophytic tree Duroia hirsuta (Rubiaceae) and its associated, minute, non-aggressive Myrmelachista sp. (Formicinae) ants is poorly investigated, even though the small understory tree is surrounded by a conspicuos zone almost devoid of vegetation. As to the cause of these barely vegetated "Devil's Gardens", two alternative interpretations exist in the literature: the pruning activity of ants and allelopathy. We investigated the mutualistic Duroia-Myrmelachista system in the Ecuadorian Amazon, focusing on the phenomenon of the areas of reduced vegetation, as well as on the natural history of the associated ants. We conducted planting experiments and behavioral studies, including coccoids, the third partner in this mutualism. Because the well-studied Triplaris americana (Polygonaceae) possesses analogous vegetation free surroundings created by the assiduous mechanical pruning of its mutualistic ant partner (Pseudomyrmex sp.), parallel comparative investigations of this ant-plant association were conducted. The two systems are not equivalent; it was demonstrated that the remarkably inactive Myrmelachista ants are not responsible for the bare areas around Duroia plants. Instead, we regard allelopathy as the causative agent for the clear zones.