Mariona Hernández-Mariné

High rate algal pond operating strategies for urban wastewater nitrogen removal

Two experimental high rate algal ponds (HRAPs) (1.5m2, 570 L per unit), each with a secondaryclarifier for algal biomass separation (0.025 m2,without recirculation), were fed with urban wastewaterfor a one-year period (June 1993 to July 1994). TheHRAPs were installed on the roof of the Department ofHydraulic, Coastal and Environmental Engineering ofthe Technical University of Catalonia, Barcelona,Spain (lat. 41° 24′ 42″ N; long. 2° 7′42″ E). Nitrogen removal efficiency and changes intotal nitrogen, total organic nitrogen,NH4+-N, and oxidized nitrogen underdifferent hydraulic retention times (HRTs) werecompared. HRAP A was always operated at a higherHRT than HRAP B. Both HRAPs were subjected to thesame environmental conditions of solar radiation, airtemperature and influent water quality. Grab samplesof influent, effluent of the HRAP (mixed liquor) andfinal effluent from the clarifiers were taken once aweek. The annual average nitrogen removal was 73% forHRAP A, and 57% for HRAP B. Higher removal in HRAP Awas due to a lower inorganic nitrogen concentration inits effluent. Significant differences (p> 0.05) inthe relative proportions of nitrogen forms between thetwo HRAPs were observed only in autumn and winter.This was mainly because HRAP B did not achieve a highlevel of NH4+-N removal by stripping andalgal uptake, as observed in HRAP A. NH4+-Nstripping was the most important mechanism fornitrogen removal (mean efficiency of 47% and 32% inHRAP A and B, respectively) followed by algal uptake,and subsequent algal separation in the clarifiers(mean efficiency of 26% and 25% in HRAP A and Brespectively). The conclusion of this study is thatHRT determines both the nitrogen removal efficiencyand the distribution of nitrogen forms in the effluentof a HRAP. The nitrogen removal level can becontrolled through suitable HRT operating strategies.By operating at a HRT of 4 days in spring and summer,and 10 days in autumn and winter, nitrogenconcentration in the effluent of a HRAP system can bereduced to less than 15 mg L-1 N.

Factors affecting the weathering and colonization of monuments by phototrophic microorganisms

Phototrophic microorganisms are common inhabitants of monuments. This paper reviews different aspects of their culture, ecology and deterioration mechanisms. Opportunistic species of cyanobacteria and chlorophytes, present in soils and in the air, are commonly found on the surfaces of monuments. Their growth represents a significant input of organic matter to the stone, as estimated through chlorophyll a quantification. Monuments provide unusual niches for the growth of algal communities, as in the case of black sulfated crusts, or endolithic and hypogeal niches, where more specific processes and/or communities occur.

SALINITY TOLERANCE OF DIATOMS FROM THALASSIC HYPERSALINE ENVIRONMENTS

Thirty‐four benthic diatom strains were isolated from thalassic hypersaline marine environments and their salinity tolerance characterized in growth experiments conducted at salinities ranging from 0.5% to 17.5% (weight of total salts per volume, g·100 mL−1). The results were compared with the patterns of diatom species distribution and abundance in hypersaline evaporation ponds and tidal channels of Guerrero Negro, Baja California Sur, Mexico. The isolated strains were representative of the diatom assemblages present in the saltern ponds but were less so of natural assemblages in tidal channels. In general, we found a clear decreasing trend of diatom diversity in the field and in the isolated strains with increasing salinity. With some exceptions, the upper limit of salinity tolerance in cultivated strains corresponded to their distribution in field samples. However, the relative abundance of species in the field was not correlated with growth rates achieved in culture for the same salinities. Most cultured strains exhibited extreme euryhalinity growing well from brackish to hypersaline conditions, but the particulars of salt tolerance were quite diverse among strains. The most halotolerant taxa, two Amphora species, Amphora cf. subacutiuscula Schoeman, Nitzschia fusiformis Grunow, and Entomoneis sp., grew well in salinities ranging from 0.5% to 15%. Three strains of Pleurosigma strigosum W. Smith that were unable to grow in salinities less than 5% total salts represent the only true halophilic diatoms ever reported. The fact that many strains displayed a remarkable halotolerance, with optimal or near‐optimal growth rates at salinities as high as three times that of seawater, implies that diatoms from hypersaline environments are evolutionarily highly adapted to such environments.

The biogeochemical role of Actinobacteria in Altamira Cave, Spain

The walls and ceiling of Altamira Cave, northern Spain, are coated with different coloured spots (yellow, white and grey). Electron microscopy revealed that the grey spots are composed of bacteria and bioinduced CaCO(3) crystals. The morphology of the spots revealed a dense network of microorganisms organized in well-defined radial and dendritic divergent branches from the central area towards the exterior of the spot, which is coated with overlying spheroidal elements of CaCO(3) and CaCO(3) nest-like aggregates. Molecular analysis indicated that the grey spots were mainly formed by an unrecognized species of the genus Actinobacteria. CO(2) efflux measurements in rocks heavily covered by grey spots confirmed that bacteria-forming spots promoted uptake of the gas, which is abundant in the cave. The bacteria can use the captured CO(2) to dissolve the rock and subsequently generate crystals of CaCO(3) in periods of lower humidity and/or CO(2). A tentative model for the formation of these grey spots, supported by scanning electron microscopy and transmission electron microscopy data, is proposed.

Exploring the secrets of the three-dimensional architecture of phototrophic biofilms in caves

INTRODUCTION Biofilms are collectives of one or more species of microorganisms. They provide protection for growth, enabling microorganisms to survive hostile environments (Prakash et al., 2003) and are significant in sediment stabilization and construction (Golubic & Schneider, 2003). Biofilms comprise sessile microorganisms in different stages of growth; hence, they respond quickly to variable conditions (Costerton et al., 1987). When microorganisms adhere to a surface, their immobilised cells grow, replicate and secrete extracellular polymeric substances (EPS) that engulf them in a gelatinous matrix (Brading et al., 1995). The development of complex, adhered or aggregated communities plays a key role in the survival and reproductive success of the microorganisms involved. Biofilms can provide refuge for species that face

Unusual ultrastructural features in three strains of Cyanothece (cyanobacteria)

Abstract Three unicellular cyanobacterial strains (PCC 7425, PCC 8303, PCC 9308) assigned to the genus Cyanothece Komárek 1976, which showed an unusually high content of light refractile inclusions when viewed by phase-contrast microscopy, were characterized by confocal laser scanning microscopy and transmission electron microscopy. All strains had concentric cortical thylakoids and a compact central nucleoid. Frequently, the two innermost thylakoid membranes protruded to form circular enclosures containing cytoplasm or electron-transparent granules, or both. The largest granules were partially immersed in the nucleoid region, but they remained attached to the inner cortical thylakoids by a single narrow connection. The pattern of binary cell division in strain PCC 7425 was different than that in strains PCC 8303 and PCC 9308. In the former, all cell wall layers invaginated simultaneously, whereas in the latter the invagination of the outer membrane was delayed compared to that of the cytoplasmic membrane and the peptidoglycan layer. Thus, prior to completion of cell division, the new daughter cells of strains PCC 8303 and PCC 9308 were transiently connected by a thick septum, which was not observed in strain PCC 7425. Nucleoid partitioning coincided with initiation of cell division in all three strains and was unlike that reported in other bacteria and in archaea, in which separation of the nucleoids precedes cell division. Based on the common morphological and ultrastructural features, the three strains of Cyanothece examined constitute a distinct cluster, which might deserve independent generic status.

Why there is such luxurious growth in the hypogean environments

Organisms building biofilms are of considerable interest in the context of degradation of cultural heritage. Particularly, hypogean environments exposed to artificial light are colonized by microbial communities, which damage walls and frescoes. In order to ascertain the mechanisms by which phototophic biofilms thrive under the particular conditions of hypogea, the organism composition and three-dimensional structure of biofilms from the Roman catacombs St. Callistus and Domitilla were studied. The main phototrophic organisms forming the biofilms were filamentous sheathed cyanobacteria and mosses. Biofilms were spatially very heterogeneous in thickness, density and organism composition but could be classified as regards their main organisms. There was a trend of decreasing diversity in the phototrophic composition of the biofilms under lower irradiances, the one at the lowest irradiance being uniquely built by erected filaments of Leptolynghya sp. Except for this biofilm, the main organism composition was not clearly related with decreasing irradiance. However, biofilms from dim light samples were porous and the filamentous cyanobacteria in them were erected. Leplolyngbya sp., the most ubiquitous species, displayed a high number of phycobilisomes and its hormogonia a gliding movement that allowed colonization of substrata. Such mechanisms may have an important role for thriving under the low light conditions of the catacombs.

Uncovering the origin of the black stains in Lascaux cave in France

Lascaux Cave in France was discovered in 1940. Since being opened to visitors the cave has suffered three major microbial outbreaks. The current problem is the fast dissemination of black stains which are threatening the Palaeolithic paintings. Previous data pointed to the involvement of new fungal species in the formation of black stains on the rock walls and ceiling. However, it appears that there could be other reasons for the formation of different and extensive black stains coating the surface of the clayey sediments. Our analyses reveal that black stains on clayey sediments are mainly produced by Acremonium nepalense, a manganese oxide-depositing fungus, widely distributed in the cave. Thus, in Lascaux Cave, the black stains have a dual origin: on limestone rocks they are mainly produced by the accumulation of fungal melanins, and on clayey sediments by the biogenic deposition of black manganese oxides.

Morphology and molecular evaluation of Iphinoe spelaeobios gen. nov., sp. nov. and Loriellopsis cavernicola gen. nov., sp. nov., two stigonematalean cyanobacteria from Greek and Spanish caves

Caves have generally been found to host phototrophic micro-organisms from various taxonomic groups, with cyanobacteria comprising an important group that have adapted to these stable and highly specific environments. A polyphasic study based on aspects of classical morphology and molecular data revealed two new monospecific genera from fresh material of Greek and Spanish caves. Both taxa are characterized by obligatory true branching (T-type, V-type and false branching), the presence of heterocysts, and reproduction by hormocysts and akinetes. They shared some similarities in their morphological characteristics as revealed by light, scanning electron and transmission electron microscopy, but phylogenetic analysis based on 16S rRNA gene sequences showed that the two phylotypes were different (89.8% similarity); this represents an example of shared morphology in genetically different strains of cave-adapted species. Phenotypic and genetic traits strongly support classification of the phylotypes as independent taxa in the order Stigonematales (the most differentiated and complicated group of cyanobacteria), family Loriellaceae Geitl 1925. Hence, the names Iphinoe spelaeobios Lamprinou and Pantazidou gen. nov., sp. nov. and Loriellopsis cavernicola Hernández-Mariné and Canals gen. nov., sp. nov. are proposed.

Enigmatic reticulated filaments in subsurface granite.

In the last few years, geomicrobiologists have focused their researches on the nature and origin of enigmatic reticulated filaments reported in modern and fossil samples from limestone caves and basalt lava tubes. Researchers have posed questions on these filaments concerning their nature, origin, chemistry, morphology, mode of formation and growth. A tentative microbial origin has been elusive since these filaments are found as hollow tubular sheaths and could not be affiliated to any known microorganism. We describe the presence of similar structures in a 16th century granite tunnel in Porto, Northwest Portugal. The reticulated filaments we identify exhibit fine geometry surface ornamentation formed by cross-linked Mn-rich nanofibres, surrounded by a large amount of extracellular polymeric substances. Within these Mn-rich filaments we report for the first time the occurrence of microbial cells.