Antonio Camacho

Environmental distribution of prokaryotic taxa

BackgroundThe increasing availability of gene sequences of prokaryotic species in samples extracted from all kind of locations allows addressing the study of the influence of environmental patterns in prokaryotic biodiversity. We present a comprehensive study to address the potential existence of environmental preferences of prokaryotic taxa and the commonness of the specialist and generalist strategies. We also assessed the most significant environmental factors shaping the environmental distribution of taxa.ResultsWe used 16S rDNA sequences from 3,502 sampling experiments in natural and artificial sources. These sequences were taxonomically assigned, and the corresponding samples were also classified into a hierarchical classification of environments. We used several statistical methods to analyze the environmental distribution of taxa. Our results indicate that environmental specificity is not very common at the higher taxonomic levels (phylum to family), but emerges at lower taxonomic levels (genus and species). The most selective environmental characteristics are those of animal tissues and thermal locations. Salinity is another very important factor for constraining prokaryotic diversity. On the other hand, soil and freshwater habitats are the less restrictive environments, harboring the largest number of prokaryotic taxa. All information on taxa, samples and environments is provided at the envDB online database, http://metagenomics.uv.es/envDB.ConclusionsThis is, as far as we know, the most comprehensive assessment of the distribution and diversity of prokaryotic taxa and their associations with different environments. Our data indicate that we are still far from characterizing prokaryotic diversity in any environment, except, perhaps, for human tissues such as the oral cavity and the vagina.

Microbial microstratification, inorganic carbon photoassimilation and dark carbon fixation at the chemocline of the meromictic Lake Cadagno (Switzerland) and its relevance to the food web

Abstract: The microstratification of the microbial community at the chemocline of Lake Cadagno and the associated inorganic carbon fixation activity was studied by fine layer sampling. A deep chlorophyll maximum caused by diatoms overlying Cryptomonas was found at the upper edge of the chemocline. A high population density of phototrophic sulphur bacteria, mainly Amoebobacter cf. purpureus, occurred closely below the oxic-anoxic boundary. Despite the small fraction of total lake volume represented by the chemocline, half of the total carbon photoassimilation of the lake occurred within the chemocline with approximately equal contributions by oxygenic and anoxygenic phototrophs. Rates of dark carbon fixation in the chemocline were even higher than rates of photoassimilation, especially at the depths where oxygen and sulphide coexisted during part of the day. These results indicate a substantial contribution by chemolithotrophic organisms to the carbon cycle in Lake Cadagno. Analysis of stable carbon isotopes suggests that zooplankton may obtain as much as half of its carbon at the chemocline, indicating a strong link between production in anoxic waters and the food web in the oxic part of the lake.

Key roles for freshwater Actinobacteria revealed by deep metagenomic sequencing

Freshwater ecosystems are critical but fragile environments directly affecting society and its welfare. However, our understanding of genuinely freshwater microbial communities, constrained by our capacity to manipulate its prokaryotic participants in axenic cultures, remains very rudimentary. Even the most abundant components, freshwater Actinobacteria, remain largely unknown. Here, applying deep metagenomic sequencing to the microbial community of a freshwater reservoir, we were able to circumvent this traditional bottleneck and reconstruct de novo seven distinct streamlined actinobacterial genomes. These genomes represent three new groups of photoheterotrophic, planktonic Actinobacteria. We describe for the first time genomes of two novel clades, acMicro (Micrococcineae, related to Luna2,) and acAMD (Actinomycetales, related to acTH1). Besides, an aggregate of contigs belonged to a new branch of the Acidimicrobiales. All are estimated to have small genomes (approximately 1.2 Mb), and their GC content varied from 40 to 61%. One of the Micrococcineae genomes encodes a proteorhodopsin, a rhodopsin type reported for the first time in Actinobacteria. The remarkable potential capacity of some of these genomes to transform recalcitrant plant detrital material, particularly lignin‐derived compounds, suggests close linkages between the terrestrial and aquatic realms. Moreover, abundances of Actinobacteria correlate inversely to those of Cyanobacteria that are responsible for prolonged and frequently irretrievable damage to freshwater ecosystems. This suggests that they might serve as sentinels of impending ecological catastrophes.

EFFECT OF EPISODIC AND REGULAR SEWAGE DISCHARGES ON THE WATER CHEMISTRY AND MACROINVERTEBRATE FAUNA OF A MEDITERRANEAN STREAM

Physical, chemical and biological parameters were used to investigate the effects of regular and episodic sewage inputs,either domestic or industrial, on the water quality of a smallMediterranean stream (River Magro, eastern Spain). Although results from chemical analyses were useful for monitoring waterquality in areas where sewage discharge was regular, episodic andlocalised discharges from industries or farms were not detected by chemical analyses because sampling was performed seasonally. An adequately small number of macroinvertebrate taxa were foundto indicate water quality changes within R. Magro, and the presence/absence and relative abundance of these taxa can be usedas a simple, rapid, low cost method for detecting changes in water quality. Macroinvertebrates are highly sensitive to episodic sewage discharges, which are difficult to detect by classical physical and chemical monitoring.

Metagenomes of Mediterranean Coastal Lagoons

Coastal lagoons, both hypersaline and freshwater, are common, but still understudied ecosystems. We describe, for the first time, using high throughput sequencing, the extant microbiota of two large and representative Mediterranean coastal lagoons, the hypersaline Mar Menor, and the freshwater Albufera de Valencia, both located on the south eastern coast of Spain. We show there are considerable differences in the microbiota of both lagoons, in comparison to other marine and freshwater habitats. Importantly, a novel uncultured sulfur oxidizing Alphaproteobacteria was found to dominate bacterioplankton in the hypersaline Mar Menor. Also, in the latter prokaryotic cyanobacteria were almost exclusively comprised by Synechococcus and no Prochlorococcus was found. Remarkably, the microbial community in the freshwaters of the hypertrophic Albufera was completely in contrast to known freshwater systems, in that there was a near absence of well known and cosmopolitan groups of ultramicrobacteria namely Low GC Actinobacteria and the LD12 lineage of Alphaproteobacteria.

Temperature effects on carbon and nitrogen metabolism in some Maritime Antarctic freshwater phototrophic communities

Biofilms growing on ice and benthic mats are among the most conspicuous biological communities in Antarctic landscapes and harbour a high diversity of organisms. These communities are consortia that make important contributions to carbon and nitrogen input in non-marine Antarctic ecosystems. Here, we study the effect of increasing temperatures on the carbon and nitrogen metabolism of two benthic communities on Byers Peninsula (Livingston Island, Maritime Antarctica): a biofilm dominated by green algae growing on seasonal ice, and a land-based microbial mat composed mainly of cyanobacteria. Inorganic carbon photoassimilation, urea and nitrate uptake and N2-fixation (acetylene reduction activity) rates were determined in situ in parallel at five different temperatures (0, 5, 10, 15, 25°C) using thermostatic baths. The results for the cyanobacterial mat showed that photosynthesis and N2-fixation responded positively to increased temperatures, but urea and NO3− uptake rates did not show a significant variation related to temperature. This microbial mat exhibits relatively low activity at 0°C whereas at higher temperatures (up to 15°C), N2-fixation rate increased significantly. Similarly, the maximum photosynthetic activity increased in parallel with temperature and showed no saturation up to 25°C. In contrast, the ice biofilm displayed higher photosynthetic activity at 0°C than at the other temperatures assayed, and it showed elevated photoinhibition at warmer temperatures.

Summer phytoplankton assemblages across trophic gradients in hard-water reservoirs

Summer phytoplankton assemblages are described and characterised according to their prevalence in a series of hard-water reservoirs of eastern Spain that had been classified in trophic categories on OECD criteria. Distribution patterns of phytoplankton species were ordinated statistically by principal components analysis (PCA). The first component was strongly related to trophic gradient and it particularly discriminated the eutrophic and hypertrophic reservoirs. The second component segregated life-forms, so that (1), on the oligo-mesotrophic side, large dinoflagellates were separated from small centric diatoms, unicellular chrysophytes and filamentous ullotrichales and, on the eu-hypertrophic side (2), colonial greens and large desmids were separated from unicellular volvocales and small centric diatoms.The large differences between eutrophic and hypertrophic reservoirs were also clearly identified in a second PCA, in which physical and chemical factors were used with the principal components solved from the phytoplankton data. From these results, a new trophic category was discerned, for which we propose the name ‘holotrophic’. This category applies to water bodies having the following main features: (1) concentrations of chorophyll, total P and total N in the range of the hypertrophic systems, but with much higher concentrations of dissolved phosphorus and ammonia and (2) phytoplankton predominantly composed by unicellular green flagellates (Pteromonas, Chlamydomonas) and chlorococcales (Scenedesmus), without cyanobacterial blooms.

Phototrophic Fe(II)-oxidation in the chemocline of a ferruginous meromictic lake

Precambrian Banded Iron Formation (BIF) deposition was conventionally attributed to the precipitation of iron-oxides resulting from the abiotic reaction of ferrous iron (Fe(II)) with photosynthetically produced oxygen. Earliest traces of oxygen date from 2.7 Ga, thus raising questions as to what may have caused BIF precipitation before oxygenic photosynthesis evolved. The discovery of anoxygenic phototrophic bacteria thriving through the oxidation of Fe(II) has provided support for a biological origin for some BIFs, but despite reports suggesting that anoxygenic phototrophs may oxidize Fe(II) in the environment, a model ecosystem of an ancient ocean where they are demonstrably active was lacking. Here we show that anoxygenic phototrophic bacteria contribute to Fe(II) oxidation in the water column of the ferruginous sulfate-poor, meromictic lake La Cruz (Spain). We observed in-situ photoferrotrophic activity through stimulation of phototrophic carbon uptake in the presence of Fe(II), and determined light-dependent Fe(II)-oxidation by the natural chemocline microbiota. Moreover, a photoferrotrophic bacterium most closely related to Chlorobium ferrooxidans was enriched from the ferruginous water column. Our study for the first time demonstrates a direct link between anoxygenic photoferrotrophy and the anoxic precipitation of Fe(III)-oxides in a ferruginous water column, providing a plausible mechanism for the bacterial origin of BIFs before the advent of free oxygen. However, photoferrotrophs represent only a minor fraction of the anoxygenic phototrophic community with the majority apparently thriving by sulfur cycling, despite the very low sulfur content in the ferruginous chemocline of Lake La Cruz.

Contribution of deep dark fixation processes to overall CO2 incorporation and large vertical changes of microbial populations in stratified karstic lakes

We carried out a detailed study in five stratified lakes in the karstic regions of NE Spain along a redox gradient combining vertical profiles of inorganic carbon dioxide fixation and analysis of microbial (bacteria and archaea) community composition determined by 16S rRNA gene fingerprinting (DGGE), microscopic counts, and pigment analysis. High rates of non-photosynthetic (i.e., “dark”) inorganic carbon incorporation were detected mostly at deeper layers after short-term in situ incubations at noon. Significant contribution of dark CO2 incorporation was observed at the whole lake level for the single time sampling, ranging between 4 and 19% of total carbon fixation measured, and up to 31% in the case of a meromictic basin. Good agreement was found between vertical patterns in redox conditions and the different microbial diversity descriptors (DGGE band sequencing, microscopic analysis, and pigment data), showing large vertical changes in microbial community composition covering a wide range of phylogenetic diversity. Cyanobacteria, Alpha and Beta-Proteobacteria, Actinobacteria, Flavobacteria and Flectobacillaceae were the most frequently recovered groups in the DGGE from oxygenated water masses. In anoxic waters, we found Beta-Proteobacteria mostly of the Rhodoferax group, Gamma-Proteobacteria (Chromatiaceae), Delta-Proteobacteria related to different sulfate reducing bacteria, Chlorobiaceae, and anaerobic Bacteroidetes spread among the Bacteroidales, Flavobacteriales and Saprospiraceae. However, as a whole, we did not find any significant correlation between dark fixation rates and either nutrient distribution and microbial community composition in the study lakes. All of this suggests that (1) different physiologies and ecologies are simultaneously contributing to the process (2) more sensitive methods are needed and more specific compounds measured and (3) some of the non-specialist microbial populations detected may carry out carbon dioxide assimilation in the dark under in situ conditions.

Spatial Dominance and Inorganic Carbon Assimilation by Conspicuous Autotrophic Biofilms in a Physical and Chemical Gradient of a Cold Sulfurous Spring: The Role of Differential Ecological Strategies

The community composition and ecophysiological features of microbial autotrophic biofilms were studied in Fuente Podrida, a cold sulfur spring located in East Spain. We demonstrated how different ecophysiological strategies, such as resistance and/or utilization of sulfide and oxygen, light adaptation, or resistance to high water flow, allow each of the microorganisms described to efficiently colonize several areas within the environmental gradient. In the zone of the spring constantly influenced by sulfide-rich waters, biofilms were formed by purple bacteria, cyanobacteria, and filamentous colorless sulfur bacteria. Purple bacteria showed higher photosynthetic efficiency per pigment unit than cyanobacteria, although they were dominant only in anoxic areas. Two filamentous cyanobacteria, strain UVFP1 and strain UVFP2, were also abundant in the sulfide-rich area. Whereas the cyanobacterial strain UVFP2 shows a strategy based on the resistance to sulfide of oxygenic photosynthesis, strain UVFP1, additionally, has the capacity for sulfide-driven anoxygenic photosynthesis. Molecular phylogenetic analyses cluster the benthic strain UVFP1 with genus Planktothrix, but with no particular species, whereas UVFP2 does not closely cluster with any known cyanobacterial species. The colorless sulfur bacterium Thiothrix sp. extended throughout the zone in which both sulfide and oxygen were present, exhibiting its capacity for chemolithoautotrophic dark carbon fixation. Downstream from the source, where springwater mixes with well-oxygenated stream water and sulfide disappears, autotrophic biofilms were dominated by diatoms showing higher photosynthetic rates than cyanobacteria and, by a lesser extent, by a sulfide-sensitive cyanobacterium (strain UVFP3) well adapted to low light availability, although in the areas of higher water velocity far from the river shore, the dominance shifted to crust-forming cyanobacteria. Both types of microorganisms were highly sensitive to sulfide impeding them from occupying sulfide-rich areas of the spring. Sulfide, oxygen, light availability, and water velocity appear as the main factors structuring the autotrophic community of Fuente Podrida spring.