Fungal treatment of agricultural washing wastewater: Comparison between two operational strategies.

Agricultural washing wastewater (AWW) is an important source of pesticides that, given its intrinsic characteristics, has a high potential to be treated by fungal bioremediation using white rot fungi. In the present study, two AWW treatment strategies were compared: a fluidized-bed reactor (FBR) with T. versicolor pellets and a rotating drum bioreactor (RDB) with T. versicolor immobilized on wood. The RDB effluent showed better results in all studied parameters compared to those of the FBR, including pesticide removal (87%), toxicity, laccase activity, COD, absorbance and microbial communities. Additionally, the fungal assemblage showed that T. versicolor was successfully immobilized in the RDB, which triggered a major shift in the initial community. Afterwards, solid by-products were treated in a fungal biopile-like system reaching high biodegradation rates. Therefore, this study validates the fungal RDB as a viable alternative for AWW treatment, opening up the possibility of a further in-situ and full-scale application.

Fungal bioremediation of agricultural wastewater in a long-term treatment: biomass stabilization by immobilization strategy.

Fungal bioremediation emerges as an effective technology for pesticide treatment, but its successful implementation depends on overcoming the problem of microbial contamination. In this regard, fungal immobilization on wood seems to be a promising strategy, but there are two main drawbacks: the predominant removal of pesticides by sorption and fungal detachment. In this study, agricultural wastewater with pesticides was treated by Trametes versicolor immobilized on wood chips in a rotary drum bioreactor (RDB) for 225 days, achieving fungal consolidation and high pesticide biodegradation through two main improvements: the use of a more favorable substrate and the modification of operating conditions. Fungal community dynamic was assessed by denaturing gradient gel electrophoresis (DGGE) analysis and subsequent prominent band sequencing, showing a quite stable community in the RDB, mainly attributed to the presence of T. versicolor. Pesticide removals were up to 54 % diuron and 48 % bentazon throughout the treatment. Afterwards, pesticide-contaminated wood chips were treated by T. versicolor in a solid biopile-like system. Hence, these results demonstrate that the microbial contamination constraint has definitely been overcome, and fungal bioremediation technology is ready to be implemented on a larger scale.

Long-term continuous treatment of non-sterile real hospital wastewater by Trametes versicolor.

BACKGROUND: Hospital wastewater is commonly polluted with high loads of pharmaceutically active compounds, which pass through wastewater treatment plants (WWTPs) and end up in water bodies, posing ecological and health risks. White-rot fungal treatments can cope with the elimination of a wide variety of micropollutants while remaining ecologically and economically attractive. Unfortunately, bacterial contamination has impeded so far a successful implementation of fungal treatment for real applications. RESULTS: This work embodied a 91-day long-term robust continuous fungal operation treating real non-sterile hospital wastewater in an air pulsed fluidized bed bioreactor retaining the biomass. The hydraulic retention time was 3 days and the ageing of the biomass was avoided through partial periodic biomass renovation resulting in a cellular retention time of 21 days. Evolution of microbial community and Trametes abundance were evaluated. CONCLUSIONS: The operation was able to maintain an average pharmaceutical load removal of over 70% while keeping the white-rot fungus active and predominant through the operation.

Are dominant microbial sub-surface communities affected by water quality and soil characteristics?

Subsurface microorganisms must deal with quite extreme environmental conditions. The lack of light, oxygen, and potentially nutrients are the main environmental stresses faced by subsurface microbial communities. Likewise, environmental disruptions providing an unbalanced positive input of nutrients force microorganisms to adapt to varying conditions, visible in the changes in microbial community diversity. In order to test microbial community adaptation to environmental changes, we performed a study in a surface Managed Aquifer Recharge facility, consisting of a settlement basin (two-day residence time) and an infiltration pond. Data on groundwater hydrochemistry, soil texture, and microbial characterization was compiled from surface water, groundwater, and soil samples at two distinct recharge operation conditions. Multivariate statistics by means of Principal Component Analysis (PCA) was the technique used to map the relevant dimensionality reduced combinations of input variables that properly describe the system behavior. The methodology selected allows including variables of different nature and displaying very different range values. Strong differences in the microbial assemblage under recharge conditions were found, coupled to hydrochemistry and grain-size distribution variables. Also, some microbial groups displayed correlations with either carbon or nitrogen cycles, especially showing abundant populations of denitrifying bacteria in groundwater. A significant correlation was found between Methylotenera mobilis and the concentrations of NO3 and SO4, and also between Vogesella indigofera and the presence of DOC in the infiltrating water. Also, microbial communities present at the bottom of the pond correlated with representative descriptors of soil grain size distribution.

Effect of cultivation conditions on ß-estradiol removal in laboratory and pilot-plant photobioreactors by an algal-bacterial consortium treating urban wastewater.

The use of microalgal consortia for urban wastewater treatment is an increasing trend, as it allows simultaneous nutrient removal and biomass production. Emerging contaminants proposed for the list of priority substances such as the hormone 17ß-estradiol are commonly found in urban wastewater, and their removal using algal monocultures has been accomplished. Due to the inherent potential of algae-based systems, this study aimed to assess the capability of native photobioreactor biomass to remove 17ß-estradiol under indoor and outdoor conditions. At the same time, the microbial community changes in regular and bioaugmented operations with Scenedesmus were assessed. The results show that almost complete removal (>93.75%) of the hormone 17ß-estradiol can be attained in the system under favourable seasonal conditions, although these conditions greatly influence biomass concentrations and microbial diversity. Even under the harsh conditions of low temperatures and solar irradiation, the established consortium removed more than 50% of the pollutant in 24 h. While species from genus Chlorella were stable during the entire operation, the microbial diversity analysis revealed that assorted and evenly distributed populations stimulate the removal rates. Bioaugmentation assays proved that the input of additional biomass results in higher overall removal and decreases the yield per mg of biomass.

Molecular and carbon isotopic characterization of an anaerobic stable enrichment culture containing Dehalobacterium sp. during dichloromethane fermentation.

Biodegradation of dichloromethane (DCM) under reducing conditions is of major concern due to its widespread detection in contaminated groundwaters. Here, we report an anaerobic enrichment culture derived from a membrane bioreactor operating in an industrial wastewater treatment plant, capable of fermenting DCM and the brominated analogue dibromomethane (DBM). Comparative analysis of bacterial 16S rDNA-DGGE profiles from fresh liquid medium inoculated with single colonies picked from serial dilution-to-extinction agar vials showed that cultures degrading DCM contained a predominant band belonging to Dehalobacterium, however this band was absent in cultures unable to degrade DCM. Analysis of the microbial composition of the enrichment by bacterial 16S rRNA gene amplicon paired-end sequencing confirmed the presence of Dehalobacterium together with three additional phylotypes belonging to Acetobacterium, Desulfovibrio, and Wolinella, representing all four operational taxonomic units >99.9% of the retrieved sequences. The carbon isotopic fractionation (ε) determined for DCM degradation in this culture was -27±2‰. This value differs from the ε previously reported for the DCM-fermentative bacteria Dehalobacter (-15.5±1.5‰) but they are both significantly different from those reported for facultative methylotrophic organisms (ranging from -45 to -61‰). This significant difference in the ε allows differentiating between hydrolytic transformation of DCM via glutathione-dependent dehalogenases and fermentation pathway. CAPSULE: The carbon isotopic fractionation of dichloromethane by an enriched Dehalobacterium-containing culture has significant potential to monitor biodegradation of DCM in groundwaters.

Characterization of Soil Suppressiveness to Root-Knot Nematodes in Organic Horticulture in Plastic Greenhouse.

The fluctuation of Meloidogyne population density and the percentage of fungal egg parasitism were determined from July 2011 to July 2013 in two commercial organic vegetable production sites (M10.23 and M10.55) in plastic greenhouses, located in northeastern Spain, in order to know the level of soil suppressiveness. Fungal parasites were identified by molecular methods. In parallel, pot tests characterized the level of soil suppressiveness and the fungal species growing from the eggs. In addition, the egg parasitic ability of 10 fungal isolates per site was also assessed. The genetic profiles of fungal and bacterial populations from M10.23 and M10.55 soils were obtained by Denaturing Gradient Gel Electrophoresis (DGGE), and compared with a non-suppressive soil (M10.33). In M10.23, Meloidogyne population in soil decreased progressively throughout the rotation zucchini, tomato, and radish or spinach. The percentage of egg parasitism was 54.7% in zucchini crop, the only one in which eggs were detected. Pochonia chlamydosporia was the only fungal species isolated. In M10.55, nematode densities peaked at the end of the spring-summer crops (tomato, zucchini, and cucumber), but disease severity was lower than expected (0.2-6.3). The percentage of fungal egg parasitism ranged from 3 to 84.5% in these crops. The results in pot tests confirmed the suppressiveness of the M10.23 and M10.55 soils against Meloidogyne. The number of eggs per plant and the reproduction factor of the population were reduced (P < 0.05) in both non-sterilized soils compared to the sterilized ones after one nematode generation. P. chlamydosporia was the only fungus isolated from Meloidogyne eggs. In in vitro tests, P. chlamydosporia isolates were able to parasitize Meloidogyne eggs from 50 to 97% irrespective of the site. DGGE fingerprints revealed a high diversity in the microbial populations analyzed. Furthermore, both bacterial and fungal genetic patterns differentiated suppressive from non-suppressive soils, but the former showed a higher degree of similarity between both suppressive soils than the later.

Organic amendments to avocado crops induce suppressiveness and influence the composition and activity of soil microbial communities.

One of the main avocado diseases in southern Spain is white root rot caused by the fungus Rosellinia necatrix Prill. The use of organic soil amendments to enhance the suppressiveness of natural soil is an inviting approach that has successfully controlled other soilborne pathogens. This study tested the suppressive capacity of different organic amendments against R. necatrix and analyzed their effects on soil microbial communities and enzymatic activities. Two-year-old avocado trees were grown in soil treated with composted organic amendments and then used for inoculation assays. All of the organic treatments reduced disease development in comparison to unamended control soil, especially yard waste (YW) and almond shells (AS). The YW had a strong effect on microbial communities in bulk soil and produced larger population levels and diversity, higher hydrolytic activity and strong changes in the bacterial community composition of bulk soil, suggesting a mechanism of general suppression. Amendment with AS induced more subtle changes in bacterial community composition and specific enzymatic activities, with the strongest effects observed in the rhizosphere. Even if the effect was not strong, the changes caused by AS in bulk soil microbiota were related to the direct inhibition of R. necatrix by this amendment, most likely being connected to specific populations able to recolonize conducive soil after pasteurization. All of the organic amendments assayed in this study were able to suppress white root rot, although their suppressiveness appears to be mediated differentially.

Diversity of the bacterial community in the surface soil of a pear orchard based on 16S rRNA gene analysis.

A cultivation-independent approach based on polymerase chain reaction (PCR)-amplified partial small subunit rRNA genes was used to characterize bacterial populations in the surface soil of a commercial pear orchard consisting of different pear cultivars during two consecutive growing seasons. Pyrus communis L. cvs Blanquilla, Conference, and Williams are among the most widely cultivated cultivars in Europe and account for the majority of pear production in Northeastern Spain. To assess the heterogeneity of the community structure in response to environmental variables and tree phenology, bacterial populations were examined using PCR-denaturing gradient gel electrophoresis (DGGE) followed by cluster analysis of the 16S ribosomal DNA profiles by means of the unweighted pair group method with arithmetic means. Similarity analysis of the band patterns failed to identify characteristic fingerprints associated with the pear cultivars. Both environmentally and biologically based principal-component analyses showed that the microbial communities changed significantly throughout the year depending on temperature and, to a lesser extent, on tree phenology and rainfall. Prominent DGGE bands were excised and sequenced to gain insight into the identities of the predominant bacterial populations. Most DGGE band sequences were related to bacterial phyla, such as Bacteroidetes, Cyanobacteria, Acidobacteria, Proteobacteria, Nitrospirae, and Gemmatimonadetes, previously associated with typical agronomic crop environments.

Diversity of the bacterial community in the surface soil of a pear orchard based on 16S rRNA gene analysis

A cultivation-independent approach based on polymerase chain reaction (PCR)-amplified partial small subunit rRNA genes was used to characterize bacterial populations in the surface soil of a commercial pear orchard consisting of different pear cultivars during two consecutive growing seasons. Pyrus communis L. cvs Blanquilla, Conference, and Williams are among the most widely cultivated cultivars in Europe and account for the majority of pear production in Northeastern Spain. To assess the heterogeneity of the community structure in response to environmental variables and tree phenology, bacterial populations were examined using PCR-denaturing gradient gel electrophoresis (DGGE) followed by cluster analysis of the 16S ribosomal DNA profiles by means of the unweighted pair group method with arithmetic means. Similarity analysis of the band patterns failed to identify characteristic fingerprints associated with the pear cultivars. Both environmentally and biologically based principal-component analyses showed that the microbial communities changed significantly throughout the year depending on temperature and, to a lesser extent, on tree phenology and rainfall. Prominent DGGE bands were excised and sequenced to gain insight into the identities of the predominant bacterial populations. Most DGGE band sequences were related to bacterial phyla, such as Bacteroidetes, Cyanobacteria, Acidobacteria, Proteobacteria, Nitrospirae, and Gemmatimonadetes, previously associated with typical agronomic crop environments (AU)

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Microcosm experiments of oil degradation by microbial mats. II. The changes in microbial species.

The influence of microbial mats on the degradation of two crude oils (Casablanca and Maya) and the effect of oil pollution on the mat structure were assessed using model ecosystems, prepared under laboratory conditions subject to tidal movements, from pristine Ebro Delta microbial-mat ecosystems. Both selected oils are examples of those currently used for commercial purposes. Casablanca crude oil is aliphatic with a low viscosity; Maya represents a sulphur-rich heavy crude oil that is predominantly aromatic. In the unpolluted microcosms, Microcoleus chthonoplastes-, Phormidium- and Oscillatoria-like were the dominant filamentous cyanobacterial morphotypes, whilst Synechoccocus-, Synechocystis- and Gloeocapsa-like were the most abundant unicellular cyanobacteria. After oil contamination, no significant changes of chlorophyll a and protein concentrations were observed, though cyanobacterial diversity shifts were monitored. Among filamentous cyanobacteria, M. chthonoplastes-like morphotype was the most resistant for both oils, unlike the other cyanobacteria, which tolerated Casablanca but not Maya. Unicellular cyanobacteria seemed to be resistant to pollution with both essayed oils, with the exception of the morphotype resembling Gloeocapsa, which was sensitive to both oils. The crude-oil addition also had a significant effect on certain components of the heterotrophic microbial community. Casablanca oil induced an increase in anaerobic heterotrophic bacteria, whereas the opposite effect was observed in those heterotrophs when polluted with Maya oil. The overall results, microbiological and crude-oil transformation analysis, indicate that the indigenous community has a considerable potential to degrade oil components by means of the metabolic cooperation of phototrophic and heterotrophic populations.

Spatial heterogeneity of bacterial populations in monomictic Lake Estanya (Huesca, Spain).

Bacterial population changes were investigated in the monomictic Lake Estanya by combining microscopic analysis and two molecular methods involving the amplification of 16S rDNA genes using primers for the domain Bacteria and subsequent restriction fragment length polymorphism (PCR-RFLP) and denaturing gradient gel electrophoresis (PCR-DGGE). Both approaches revealed the vertical distribution of predominant microbial morphotypes and phylotypes in both holomictic and stratified periods, respectively, and showed that variations in structure and composition of bacterial populations are occurring in this lake as a function of depth and time. Through principal component analysis (PCA), these shifts could be related to different physicochemical parameters with temperature, oxygen concentration, and the incident light being of paramount importance as structuring variables. Comparison of RFLP and DGGE profiles by scoring similarities using the Jaccard coefficient and then building a multidimensional scaling map (MDS) showed equivalent results. Both techniques revealed that bacterial populations, present in the whole water column in the holomictic period, showed a high similarity with those located in the deeper part of the lake in the stratified period, evidencing that other factors, both biotic and abiotic, should also be considered as a force driving change in the composition of the bacterial community. Furthermore, DGGE analysis showed that sequences from prominent bands were affiliated to members of four major phyla of the domain Bacteria: Cyanobacteria, Bacteroidetes, Proteobacteria, and Actinobacteria, most of which corresponded to heterotrophic bacterial populations involved in carbon, sulfide, and nitrogen biogeochemical cycles, which were indistinguishable under the light microscope.

Diversity of anoxygenic phototrophic sulfur bacteria in the microbial mats of the Ebro Delta: a combined morphological and molecular approach.

The diversity of purple and green sulfur bacteria in the multilayered sediments of the Ebro Delta was investigated. Specific oligonucleotide primers for these groups were used for the selective amplification of 16S rRNA gene sequences. Subsequently, amplification products were separated by denaturing gradient gel electrophoresis and sequenced, which yielded a total of 32 sequences. Six of the sequences were related to different cultivated members of the green sulfur bacteria assemblage, whereas seven fell into the cluster of marine or halophilic Chromatiaceae. Six sequences were clustered with the family Ectothiorhodospiraceae, three of the six being closely related to chemotrophic bacteria grouped together with Halorhodospira genus, and the other three forming a group related to the genus Ectothiorhodospira. The last thirteen sequences constituted a cluster where no molecular isolate from microbial mats has so far been reported. Our results indicate that the natural diversity in the ecosystem studied has been significantly underestimated in the past and point out the presence of novel species not related to all known purple sulfur bacteria. Furthermore, the detection of green sulfur bacteria, after only an initial step of enrichment, suggests that -- with the appropriate methodology -- several genera, such as Prosthecochloris, could be established as regular members of marine microbial mats.

Seasonal microbial ribotype shifts in the sulfurous karstic lakes Cisó and Vilar, in northeastern Spain / Cambios estacionales en los ribotipos microbianos en los lagos cársticos y sulfurosos Cisó y Vilar, en el nordeste de España

Spatio-temporal changes in two sulfurous lakes from the karstic area of Banyoles (Girona, Spain), holomictic lake Cisó and meromictic lake Vilar, were studied over one year. Samples were collected at different depths from the two lakes on the same days, during each of the four seasons, and several physico-chemical variables (temperature, light, pH, conductivity, sulfide, oxygen concentration, pigment concentrations, etc.) were measured. To fingerprint bacterial populations from each sample, DNA was extracted, bacterial 16S rRNA genes were amplified by PCR, and restriction fragment length polymorphism (RFLP) analyses of the total bacterial 16S rDNAs were performed. Each 16S rDNA pool was independently digested with three restriction endonucleases (AluI, HinfI, and RsaI) and separated electrophoretically. More restriction fragments were obtained from the Lake Vilar samples than from the Lake Cisó samples. Moreover, intrasample and intersample differences were observed in each lake. RFLP patterns were compared by scoring similarities using the Jaccard coefficient and then building a multidimensional scaling (MDS) map from the resulting similarities matrix. In both lakes, results indicated that seasonality was mostly responsible for the observed fluctuations in the RFLP patterns, while the effect of stratification was less pronounced (AU)

Se estudió a lo largo de un año el cambio espacio-temporal que se produjo en dos lagos sulfurosos de la zona cárstica de Banyoles (Girona, España), el lago Cisó, holomíctico, y el Lago Vilar, meromíctico. Se tomaron muestras a diferentes profundidades en los dos lagos los mismos días durante las cuatro estaciones y se midieron algunas variables fisicoquímicas (temperatura, luz, pH, conductividad, sulfuro, concentraciones de oxígeno y de pigmentos, etc.). Para obtener la impronta genética de las poblaciones bacterianas de cada muestra, se extrajo el DNA, se amplificaron los genesdel 16S rRNA mediante PCR y se analizó el polimorfismo en la longitud de los fragmentos de restricción (RFLP) del total de 16S rDNA bacteriano. Los diferentes conjuntos de 16S rDNA bacteriano fueron digeridos de manera independiente con tres endonucleasas de restricción (AluI, HinfI, y RsaI) y separados por electroforesis. Se obtuvieron más fragmentos de restricción de las muestras del lago Vilar que del Cisó. Además, en cada lago se observaron también diferencias dentro de cada muestra y entre las diferentes muestras. Luego se compararon los patrones de RFLP puntuando las similitudes mediante el coeficiente Jaccard y la creación un mapa de escalamiento multidimensional (MDS) a partir de la matriz de similitudes resultante. Los resultados indicaron que la estacionalidad era la principal causa de las fluctuaciones observadas en los patrones de RFLP en ambos lagos, mientras que el efecto de la estratificación era menos pronunciado (AU)

Seasonal microbial ribotype shifts in the sulfurous karstic lakes Cisó and Vilar, in northeastern Spain.

Spatio-temporal changes in two sulfurous lakes from the karstic area of Banyoles (Girona, Spain), holomictic lake Cisó and meromictic lake Vilar, were studied over one year. Samples were collected at different depths from the two lakes on the same days, during each of the four seasons, and several physico-chemical variables (temperature, light, pH, conductivity, sulfide, oxygen concentration, pigment concentrations, etc.) were measured. To fingerprint bacterial populations from each sample, DNA was extracted, bacterial 16S rRNA genes were amplified by PCR, and restriction fragment length polymorphism (RFLP) analyses of the total bacterial 16S rDNAs were performed. Each 16S rDNA pool was independently digested with three restriction endonucleases (AluI, HinfI, and RsaI) and separated electrophoretically. More restriction fragments were obtained from the Lake Vilar samples than from the Lake Cisó samples. Moreover, intrasample and intersample differences were observed in each lake. RFLP patterns were compared by scoring similarities using the Jaccard coefficient and then building a multidimensional scaling (MDS) map from the resulting similarities matrix. In both lakes, results indicated that seasonality was mostly responsible for the observed fluctuations in the RFLP patterns, while the effect of stratification was less pronounced.

Distribution of phototrophic populations and primary production in a microbial mat from the Ebro Delta, Spain.

Microbial mats arising in the sand flats of the Ebro Delta (Tarragona, Spain) were investigated during the summer season, when the community was highly developed. These mats are composed of three pigmented layers of phototrophic organisms, an upper brown layer mainly composed of Lyngbya aestuarii and diatoms, an intermediate green layer of the cyanobacterium Microcoleus chthonoplastes, and an underlying pink layer of a so-far unidentified purple sulfur bacterium. In the photic zone, oxygenic phototrophs constitute about 58% of total photosynthetic biomass, measured as biovolume, and anoxygenic phototrophs represent 42%. Diatoms constitute 11.8% of the oxygenic biomass, M. chthonoplastes 61.2%, and L. aestuarii and coccoid cyanobacteria 20.6 and 6.4%, respectively. In this laminated community, organic matter has an autochthonous origin, and photosynthesis is the most important source of organic carbon. Oxygen production reaches up to 27.2 mmol O(2) m(-2) h(-1), measured at 1000 microE m(-2) s(-1) light intensity, whereas oxidation of sulfide in the light has been calculated to be 18.6 mmol S m(-2) h(-1). This amount represents 26% of the total photosynthetic production in terms of photoassimilated carbon, demonstrating the important role of anoxygenic phototrophs as primary producers in the pink layer of Ebro Delta microbial mats.

Factors affecting interpretation of restriction fragment length polymorphism (RFLP) patterns from PCR-amplified bacterial 16S rRNA genes: operon number and primer mismatching.

PCR methods have been shown to be biased by several factors. In the present study, we have developed a theoretic and practical approximation to elucidate how the presence of mismatches at the primers annealing regions and the different number of rDNA operons per cell can influence PCR and subsequent restriction fragment length polymorphism (RFLP) analyses from bacterial populations. We have performed RFLP analyses of 16S rRNA genes amplified by PCR from mixed bacterial cultures showing different primer identities and number of rDNA operons. Our results clearly corroborate that both factors, number of rDNA operons and primers identity, clearly influence the 16S rDNA-RFLP estimations. It has been demonstrated that a higher number of operons leads to a higher efficiency of detection, but a lower degree of primer complementarity implies a decrease in such efficiency.

Distribution of phototrophic populations and primary production in a microbial mat from the Ebro Delta, Spain / Distribución de poblaciones fotótrofas y producción primaria en un tapete microbiano del delta del Ebro (España)

Microbial mats arising in the sand flats of the Ebro Delta (Tarragona, Spain) were investigated during the summer season, when the community was highly developed. These mats are composed of three pigmented layers of phototrophic organisms, an upper brown layer mainly composed of Lyngbya aestuarii and diatoms, an intermediate green layer of the cyanobacterium Microcoleus chthonoplastes, and an underlying pink layer of a so-far unidentified purple sulfur bacterium. In the photic zone, oxygenic phototrophs constitute about 58% of total photosynthetic biomass, measured as biovolume, and anoxygenic phototrophs represent 42%. Diatoms constitute 11.8% of the oxygenic biomass, M. chthonoplastes 61.2%, and L. aestuarii and coccoid cyanobacteria 20.6 and 6.4%, respectively. In this laminated community, organic matter has an autochthonous origin, and photosynthesis is the most important source of organic carbon. Oxygen production reaches up to 27.2 mmol O(2) m(-2) h(-1), measured at 1000 microE m(-2) s(-1) light intensity, whereas oxidation of sulfide in the light has been calculated to be 18.6 mmol S m(-2) h(-1). This amount represents 26% of the total photosynthetic production in terms of photoassimilated carbon, demonstrating the important role of anoxygenic phototrophs as primary producers in the pink layer of Ebro Delta microbial mats (AU)

Los tapetes microbianos que se establecen en los sedimentos litorales del delta del Ebro (Tarragona, España) fueron investigados durante el verano, cuando la comunidad estaba muy desarrollada. Dichos tapetes se componen de tres capas pigmentadas, con diferentes organismos fotótrofos. La capa superior es de color marrón y está compuesta principalmente por Lyngbya aestuarii y diatomeas. Debajo de ésta, se observa una capa intermedia de color verde, donde predomina la cianobacteria Microcoleus chthonoplastes. Finalmente, por debajo de las dos anteriores se ve una lámina rosa, en la que el organismo fototrófico dominante es una nueva bacteria roja del azufre no identificada hasta este momento. En la zona fótica, los organismos fototróficos oxigénicos representan un 58 por ciento de la biomasa fotosintética total, medida ésta como biovolumen; el 48 por ciento restante corresponde a los organismos fotótrofos anoxigénicos. En relación a la biomasa oxigénica, las diatomeas constituyen un 11,8 por ciento del total, mientras que M. chthonoplastes, L. aestuarii y las cianobacterias cocoides representan un 61,2 por ciento, un 20,6 por ciento y un 6,4 por ciento, respectivamente. En esta comunidad multilaminada, la materia orgánica es de origen autóctono y la fotosíntesis es la principal fuente de carbono orgánico. La producción de oxígeno alcanza los 27,2 mmol O2 m-2 h-1 medida a una intensidad de luz de 1000 µE m-2 s-1. Mientras que la oxidación de sulfuro a la luz es de 18,6 mmoles S m-2 h-1. Esta última cantidad representa un 26 por ciento de la producción fotosintética total, expresada como C fotoasimilado, lo cual pone de manifiesto el papel destacado de las bacterias fototrofas anoxigénicas como productores primarios en la capa roja de los ecosistemas estudiados (AU)

Seasonal population changes in the restriction fragment length polymorphism (RFLP) patterns from PCR-amplified 16S rRNA genes of predominant ribotypes in microbial mat samples from the Ebro Delta (Spain).

The stratified benthic microbial communities, which developed as a consequence of the physico-chemical gradients and the physiology of the inhabiting microorganisms, from Ebro Delta microbial mats were analyzed. 16S rRNA eubacterial and archaeal genes were amplified by polymerase chain reaction (PCR). PCR products were separately digested with three different restriction enzymes (AluI, HinfI, and RsaI) and later separated by horizontal electrophoresis to determine whether changes of predominant ribotypes are occurring over a period of a year. Comparison of total restriction patterns was performed by scoring similarities by using the Jaccard coefficient and then building a multidimensional scaling (MDS) map from the resulting similarities matrix. The three enzymes gave a consistent result, a seasonal distribution instead of a spatial and/or physiological one.