Carles A. Abella

THE MOLAR EXTINCTION COEFFICIENT OF BACTERIOCHLOROPHYLL E AND THE PIGMENT STOICHIOMETRY IN CHLOROBIUM PHAEOBACTEROIDES

We have determined the molar extinction coefficient of bacteriochlorophyll (BChl) e, the main light-harvesting pigment from brown-coloured photosynthetic sulfur bacteria. The extinction coefficient was determined using pure [Pr,E]BChl eF isolated by reversed-phase HPLC from crude pigment extracts of Chlorobium (Chl.) phaeobacteroides strain CL1401. The extinction coefficients at the Soret and Qy bands were determined in four organic solvents. The extinction coefficient of BChl e differs from those of other related Chlorobium chlorophylls (BChl c and BChl d) but is similar to that of chlorophyll b. The determined extinction coefficient was used to calculate the stoichiometric BChl e to BChl a and BChl e to carotenoids ratios in whole cells and isolated chlorosomes from Chl. phaeobacteroides strain CL1401 using the spectrum-reconstruction method (SRCM) described by Naqvi et al. (1997) (Spectrochim Acta A Mol Biomol Spectrosc 53: 2229–2234) . In isolated chlorosomes, BChl a content was ca. 1% of the total BChl content and the stoichiometric ratio of BChl e to carotenoids was 6. In whole cells, however, BChl a content was 3–4%, owing to the presence of BChl a-containing elements, i.e. FMO protein and reaction centre. An average of 5 BChl e molecules per carotenoid was determined in whole cells.

Light-harvesting adaptations of planktonic phototrophic micro-organisms to different light quality conditions

The effects of light spectral distribution on the composition of phototrophic microbial communities were analyzed in three metalimnetic levels (relative depth positions) of 41 lakes. Principal Component Analysis was used to compare light quality conditions reaching the populations of phototrophic micro-organisms containing different photosynthetic pigments. Results allowed to identify the optimal light quality conditions for the selection of each microbial group at their respective levels. Two general light-harvesting adaptations were defined, according to the wavebands that could be related to the selection of these microbial groups. The micro-organisms adapted to use red and near-infrared light – eukaryotic phytoplankton, Chloronema spp. and green-coloured Chlorobiaceae – predominated at shallow depths (specially in waters containing high gilvin contents) using their respective Qy absorption bands. The micro-organisms adapted to green-yellow light – phycoerythrin-containing cyanobacteria, Chromatiaceae and brown-coloured Chlorobiaceae – were dominant in deep metalimnetic communities. Laboratory experiments with cultures of Chlorobium limicola and C. phaeobacteroides growing under different light quality conditions showed that the green-coloured species had higher photosynthetic activity under red light, while the brown-coloured species was more active under green light. These results demonstrated that physiological differences between micro-organisms with different light-harvesting adaptations are responsible of their selection under different light quality conditions. This selection is experimented by Chlorobiaceae (as it was previously indicated by other investigators) at the deepest positions of the metalimnetic communities (level 3), but also by Chromatiaceae and Chloronema spp. at level 2 and by the eukaryotic phytoplankton and cyanobacteria at level 1.

Mucosa-associated Faecalibacterium prausnitzii phylotype richness is reduced in patients with inflammatory bowel disease.

ABSTRACT Faecalibacterium prausnitzii depletion in intestinal diseases has been extensively reported, but little is known about intraspecies variability. This work aims to determine if subjects with gastrointestinal disease host mucosa-associated F. prausnitzii populations different from those hosted by healthy individuals. A new species-specific PCR-denaturing gradient gel electrophoresis (PCR-DGGE) method targeting the 16S rRNA gene was developed to fingerprint F. prausnitzii populations in biopsy specimens from 31 healthy control (H) subjects and 36 Crohns disease (CD), 23 ulcerative colitis (UC), 6 irritable bowel syndrome (IBS), and 22 colorectal cancer (CRC) patients. The richness of F. prausnitzii subtypes was lower in inflammatory bowel disease (IBD) patients than in H subjects. The most prevalent operational taxonomic units (OTUs) consisted of four phylotypes (OTUs with a 99% 16S rRNA gene sequence similarity [OTU99]), which were shared by all groups of patients. Their distribution and the presence of some disease-specific F. prausnitzii phylotypes allowed us to differentiate the populations in IBD and CRC patients from that in H subjects. At the level of a minimum similarity of 97% (OTU97), two phylogroups accounted for 98% of the sequences. Phylogroup I was found in 87% of H subjects but in under 50% of IBD patients (P = 0.003). In contrast, phylogroup II was detected in >75% of IBD patients and in only 52% of H subjects (P = 0.005). This study reveals that even though the main members of the F. prausnitzii population are present in both H subjects and individuals with gut diseases, richness is reduced in the latter and an altered phylotype distribution exists between diseases. This approach may serve as a basis for addressing the suitability of F. prausnitzii phylotypes to be quantified as a putative biomarker of disease and depicting the importance of the loss of these subtypes in disease pathogenesis.

Comparative study of two meromictic basins of Lake Banyoles (Spain) with sulphur phototrophic bacteria

The annual limnological dynamics of two meromictic basins of Lake Banyoles (C-III and C-IV) have been studied and compared on the basis of their physical, chemical and biological characters. Stability values calculated for both basins gave 865 g cm cm−2 and 495 g cm cm−2 for C-III and C-IV respectively. These values are in agreement with the fact that C-IV was almost completely mixed during winter. In this basin, during stratification, the monimolimnion increased in thickness as the stability increased. Isolation of the respective monimolimnia resulted in the development of anoxic conditions and the accumulation of sulphide in both C-III and C-IV, which favoured the development of dense populations of sulfur phototrophic bacteria. The purple sulphur bacterium Chromatium minus and the green sulphur bacterium Chlorobium phaeobacteroides were identified as the main components of these photosynthetic populations. The different depths at which the O2/H2S boundary was situated in both basins (and consequently the different light intensity reaching this zone) determined the growth of these bacteria. Light intensities at the chemocline of C-IV reached values up to 5% of surface incident light. In contrast, in C-III this variable was sensibly lower, with values depending on season and seldom reaching 1%. Phototrophic bacteria were consequently found earlier in C-IV than in C-III, where no significant concentrations were found until August. Finally stability is discussed as an important factor controlling chemical and biological dynamics in meromictic lakes.

Sulfide pulsing as the controlling factor of spinae production in Chlorobium limicola strain UdG 6038

Chlorobium limicola UdG 6038, a green sulfur bacterium, was isolated from anoxic sediments. Cells were gram-negative, non-motile, ovoid shaped, and contained chlorobactene and bacteriochlorophyll c as the main photosynthetic pigments. The DNA G+C content was 56.4 mol%. Ultrastructural studies revealed the presence of abundant spinae (45–110 spinae per cell) attached to the cell wall. India-ink-stained cells observed under the optical microscope were surrounded by a large capsule (5–11 μm total diameter). The presence of this capsule was coincident with the presence of a large number of spinae (> 30 spinae per cell). The mucilaginous capsule was attached to the spinae without penetrating it. In batch culture, the synthesis of spinae in strain UdG 6038 was not affected by changes in temperature, pH, salt concentration, or illumination at physiological ranges and hence, the cells remained spined. The control of spinae production was experimentally confirmed using a semicontinuous batch culture refed by sulfide pulsing. The culture remained at a low spination level (> 30 spinae per cell) only when the duration of sulfide starvation between pulses was less than 5 h. After longer sulfide starvation periods, the cells remained spined (more than 38 ± 6.3 spinae per cell). This observation supports the idea that the duration of sulfide limitation in the culture plays a key role in controlling the spination process in strain C. limicola UdG 6038. Chlorobium spinae may play an eco-physiological role in buoyancy capacity and adhesion of sulfur globules to the cells in natural environments where sulfide concentrations are expected to be highly variable.

Use of the ammonia-oxidizing bacterial-specific phylogenetic probe Nso1225 as a primer for fingerprint analysis of ammonia-oxidizer communities

Autotrophic ammonia-oxidizing bacteria (AOB) are an essential component of nitrifying wastewater treatment systems. The molecular tools used in group-specific studies are mostly based on the sequence of the 16S rRNA gene, but they have not proved to be fully specific. In this study, the sequence of the FISH probe Nso1225R was used as a reverse primer in order to analyze the AOB composition of several environmental samples by denaturing gradient gel electrophoresis (DGGE). For this purpose, samples from several environments, including aerated reactors, water treatment wetlands, and pilot plants, both aerobic and anaerobic, were analyzed. PCR fragments displayed a DGGE pattern consisting of bands melting between 30 and 40% denaturant, and a series of unresolved bands above 45%, mostly corresponding to AOB and β-non-AOB, respectively. This second set of bands corresponded to environments subjected to severe oxygen restrictions. AOB sequences showed similarity percentages higher than 92% with those of known β-AOB. Nso1225R, therefore, proved to be a good molecular phylogenetic marker for AOB samples from well-aerated systems, showing a higher specificity than the group-specific primers used previously.

Microbial ecology of planktonic filamentous phototrophic bacteria in holomictic freshwater lakes

The microbial ecology of the filamentous phototrophic bacterium, Chloronema giganteum, has been studied in the water column of three central European lakes (Schlein, Buchen and Vechten). In these lakes an anoxic layer, termed the transition zone, was located between the oxycline and the redoxcline. The migration capacity of Chloronema through this zone appears to be responsible for the natural preponderance of either straight or spiral forms. When the transition zone is less than 1 m thick the straight form is dominant, but when this transition zone is wider than 2 m the spiral form is enriched. The intermediate situations favour both filamentous forms.