Structural Interactions among Epilithic Cyanobacteria and Heterotrophic Microorganisms in Roman Hypogea

Abstract Phototrophic microbial communities present in the Roman Catacombs were characterized and different species of terrestrial epilithic cyanobacteria were found to occur as dominant organisms. Eucapsis, Leptolyngbya, Scytonema, and Fischerella were the most frequently encountered cyanobacterial taxa, while a few species of green algae and the diatom Diadesmis gallica occurred in minor amounts. Streptomyces strains, a few genera of eubacteria, and to a lesser extent fungi were always present in the same microhabitats and contributed to the deterioration of stone surfaces. The combined use of light and electron microscopy evidenced the structural relationships among rod-shaped or filamentous bacteria and cyanobacterial cells, as well as the presence of polysaccharide capsules and sheaths, and of mineral precipitates on S. julianum filaments. The significance of the intimate association among the microorganisms was discussed in relation to the damage caused by the growth of biological patinas on stone surfaces. Received: 17 February 1999; Accepted: 19 May 1999     

Ultraviolet-absorbing scytonemin and mycosporine-like amino acid derivatives in exposed, rock-inhabiting cyanobacterial lichens

 The occurrence of ultraviolet (UV)-absorbing substances like scytonemin and mycosporine-glycine is reported for the first time from cyanobacterial lichens of the genera Collema, Gonohymenia and Peltula, all coming from high-light-intensity habitats. Except for Collema with the filamentous Nostoc, all other cyanobionts belong to the unicellular genera Chroococcidiopsis, Cyanosarcina, Gloeocapsa or Myxosarcina. From transmission electron microscope studies it is evident that the pigmentation (scytonemin) is located extracellularly in the sheath of the outer thallus parts. Fluorescence microscopy and microprobe measurements clearly show UV radiation into the lichen thallus and hence the relevance of UV sunscreens for the protection of the organism. Received: 4 March 1997 / Accepted: 20 May 1997     

Characterization of Cyanobacteria by SDS-PAGE of whole-cell proteins and PCR/RFLP of the 16S rRNA gene

Planktonic, filamentous cyanobacterial strains from different genera, both toxic and nontoxic strains, were characterized by SDS-PAGE of whole-cell proteins and PCR/RFLP of the 16S rRNA gene. Total protein pattern analysis revealed the mutual relationships at the genus level. Restriction fragment length polymorphism (RFLP) of the 16S rRNA gene with reference strains proved to be a good method for the cyanobacterial taxonomy. The nonheterocystous strains outgrouped from the nitrogen-fixing ones. With both methods, Aphanizomenon clustered with Anabaena, and Nodularia with Nostoc. In the RFLP study of Anabaena, the neurotoxic strains were identical, but the hepatotoxic ones formed a heterogeneous group. Genetic distances found in the RFLP study were short, confirming that close genotypic relationships underlie considerable diversity among cyanobacterial genera. Received: 16 December 1996 / Accepted: 14 May 1997     

Sterol compositions of the filamentous nitrogen-fixing terrestrial cyanobacterium Scytonema sp

Thirteen unsaturated sterols were identified by gas chromatography--mass spectrometry using serially-coupled capillary columns from the filamentous nitrogen-fixing terrestrial cyanobacterium Scytonema sp. isolated from the microbial community of cyanobacterial on 'Black Cover' biofilms limestone walls in Jerusalem. The dominant sterols were cholest-5-en-3 beta-ol (18.9%), 3 beta-methoxycholest-5-ene (16.2%) and 3 beta-acetoxycholest-5-ene (11.2%).     

Sequential role of biosorption and biodegradation in rapid removal degradation and utilization of methyl parathion as a phosphate source by a new cyanobacterial isolate Scytonema sp. BHUS-5

A new isolate of genus Scytonema distinct from its closest relative cyanobacterium, Scytonema hofmanni was found efficient in the removal and degradation of organophosphorus (OP) pesticide, methyl parathion (MP). The cyanobacterial isolate was also capable of utilizing the phosphorus present in the MP following its degradation, which was evident from the increase in growth (chlorophyll content), biomass, protein content, and total phosphorus in comparison to cyanobacterium grown in phosphate-deficient cultures. The rapid removal of MP by the cyanobacterium during initial 6 hours of incubation was defined by the pseudo-second-order biosorption kinetics model, which indicated the involvement of chemosorption in initial removal of pesticide. Further, degradation of MP was also confirmed by the appearance of p-nitrophenol in the medium after 24 hours of incubation. Thus, the cyanobacterial isolate of Scytonema sp. BHUS-5 seems to be a potential bioremediation agent for the removal of OP pesticide, MP from the habitat.     

Controlling Harmful Cyanobacteria: Taxa-Specific Responses of Cyanobacteria to Grazing by Large-Bodied Daphnia in a Biomanipulation Scenario

Lake restoration practices based on reducing fish predation and promoting the dominance of large-bodied Daphnia grazers (i.e., biomanipulation) have been the focus of much debate due to inconsistent success in suppressing harmful cyanobacterial blooms. While most studies have explored effects of large-bodied Daphnia on cyanobacterial growth at the community level and/or on few dominant species, predictions of such restoration practices demand further understanding on taxa-specific responses in diverse cyanobacterial communities. In order to address these questions, we conducted three grazing experiments during summer in a eutrophic lake where the natural phytoplankton community was exposed to an increasing gradient in biomass of the large-bodied Daphnia magna. This allowed evaluating taxa-specific responses of cyanobacteria to Daphnia grazing throughout the growing season in a desired biomanipulation scenario with limited fish predation. Total cyanobacterial and phytoplankton biomasses responded negatively to Daphnia grazing both in early and late summer, regardless of different cyanobacterial densities. Large-bodied Daphnia were capable of suppressing the abundance of Aphanizomenon, Dolichospermum, Microcystis and Planktothrix bloom-forming cyanobacteria. However, the growth of the filamentous Dolichospermum crassum was positively affected by grazing during a period when this cyanobacterium dominated the community. The eutrophic lake was subjected to biomanipulation since 2005 and nineteen years of lake monitoring data (1996–2014) revealed that reducing fish predation increased the mean abundance (50%) and body-size (20%) of Daphnia, as well as suppressed the total amount of nutrients and the growth of the dominant cyanobacterial taxa, Microcystis and Planktothrix. Altogether our results suggest that lake restoration practices solely based on grazer control by large-bodied Daphnia can be effective, but may not be sufficient to control the overgrowth of all cyanobacterial diversity. Although controlling harmful cyanobacterial blooms should preferably include other measures, such as nutrient reductions, our experimental assessment of taxa-specific cyanobacterial responses to large-bodied Daphnia and long-term monitoring data highlights the potential of such biomanipulations to enhance the ecological and societal value of eutrophic water bodies.     

Uranium mobilization from low-grade ore by cyanobacteria

Summary Three cyanobacterial isolates (two LPP-B forms and one Anabaena or Nostoc species) from different environments could mobilize uranium from low-grade ores. After 80 days, up to 18% uranium had been extracted from coal and 51% from carbonate rock by the filamentous cyanobacterium OL3, a LPP-B form. Low growth requirements with regard to light and temperature optima make this strain a possible candidate for leaching neutral and alkaline low-grade uranium ores.     

Exopolysaccharides of Two Cyanobacterial Strains from Roman Hypogea

The cytomorphology and the exopolysaccharides of two heterocystous cyanobacteria, Scytonema ocellatum CP8-2 and Fischerella maior NAV 10 bis, isolated from biofilms collected in two Roman hypogea, were investigated from a cytochemical, ultrastructural and biochemical point of view. Cytochemical stains and observation using light and transmission electron microscopy showed a bi-layered sheath in both cyanobacteria, with a different composition in sulphated and carboxylic groups. RP-HPLC of the extracted polysaccharides forming the mucous matrix showed the neutral sugars glucose and arabinose and the charged galacturonic acid as the main monosaccharides. Circular dichroism spectra recorded at varying pH and temperature showed that these polymers followed a random coil model and did not present a structured organised conformation at ambient temperature. The outcomes of this study increase the understanding of colonization and biodeterioration of important archaeological and cultural sites by cyanobacterial biofilms and the role of the exopolymeric substances involved.     

Genome identification of the Triticum crassum complex (Poaceae) with the restriction patterns of repeated nucleotide sequences

Species of the Triticum crassum complex, tetraploid and hexaploid T. crassum, hexaploid T. juvenale, and hexaploid T. syriacum, share a similar morphology. Variation in the restriction profiles of nuclear repeated nucleotide sequences is employed in identification of genomes of these species. The data show that hexaploid T. crassum originated from hybridization of the tetraploid cytotype of T. crassum with T. tauschii. Triticum juvenale and T. syriacum originated from hybridization of tetraploid T. crassum with T. umbellulatum and T. searsii, respectively. Tetraploid T. crassum appears to be an ancient allotetraploid that originated from hybridization of primitive T. tauschii with an ancient species in the evolutionary lineage leading to the section Sitopsis of the genus Triticum.     

Genotypic and Phenotypic Diversity of Cyanobacteria in Biological Soil Crusts of the Succulent Karoo and Nama Karoo of Southern Africa

Biological soil crusts (BSCs) are communities of cryptogamic organisms, occurring in arid and semiarid regions all over the world. Based on both morphological identification and genetic analyses, we established a first cyanobacterial inventory using the biphasic approach for BSCs within two major biomes of southern Africa. The samples were collected at two different sites in the Succulent Karoo and one in the Nama Karoo. After cultivation and morphological identification, the 16S rRNA gene was sequenced from the cyanobacterial cultures. From the soil samples, the DNA was extracted, and the 16S rRNA gene sequenced. All the sequences of the clone libraries from soil and cultures were compared with those of the public databases. Forty-five different species were morphologically identified in the samples of the Succulent Karoo (observatories of Soebatsfontein and Goedehoop). Based on the genetic analyses, 60 operational taxonomic units (OTUs) were identified for the Succulent Karoo and 43 for the Nama Karoo (based on 95 % sequence similarity). The cloned sequences corresponded well with the morphologically described taxa in cultures and sequences in the public databases. Besides known species of typical crust-forming cyanobacterial genera (Microcoleus, Phormidium, Tolypothrix and Scytonema), we found sequences of so far undescribed species of the genera Leptolyngbya, Pseudanabaena, Phormidium, Oscillatoria, Schizothrix and Microcoleus. Most OTUs were restricted to distinct sites. Grazed soils showed lower taxa numbers than undisturbed soils, implying the presence of early successional crust types and reduced soil surface protection. Our combined approach of morphological identification and genetic analyses allowed both a taxa inventory and the analysis of species occurring under specific habitat conditions.     

Fungal Parasitism: Life Cycle,Dynamics and Impact on Cyanobacterial Blooms

Many species of phytoplankton are susceptible to parasitism by fungi from the phylum Chytridiomycota (i.e. chytrids). However, few studies have reported the effects of fungal parasites on filamentous cyanobacterial blooms. To investigate the missing components of bloom ecosystems, we examined an entire field bloom of the cyanobacterium Anabaena macrospora for evidence of chytrid infection in a productive freshwater lake, using a high resolution sampling strategy. A. macrospora was infected by two species of the genus Rhizosiphon which have similar life cycles but differed in their infective regimes depending on the cellular niches offered by their host. R. crassum infected both vegetative cells and akinetes while R. akinetum infected only akinetes. A tentative reconstruction of the developmental stages suggested that the life cycle of R. crassum was completed in about 3 days. The infection affected 6% of total cells (and 4% of akinètes), spread over a maximum of 17% of the filaments of cyanobacteria, in which 60% of the cells could be parasitized. Furthermore, chytrids may reduce the length of filaments of Anabaena macrospora significantly by “mechanistic fragmentation” following infection. All these results suggest that chytrid parasitism is one of the driving factors involved in the decline of a cyanobacteria blooms, by direct mortality of parasitized cells and indirectly by the mechanistic fragmentation, which could weaken the resistance of A. macrospora to grazing.     

Isolation and Characterization of Cyanobacteria from Microbial Mats of the Ebro Delta,Spain

Isolation and identification of several strains of cyanobacteria from microbial mats of the Ebro Delta, Spain, are described. A series of tenfold dilutions was the first step of isolation. Self-isolation techniques, which use one or several physiological characteristics of a cyanobacterium, were applied in some cases to obtain enrichment cultures. Twelve filamentous strains were isolated and stored in axenic culture. As only a few cyanobacterial species can be frozen and revived without any cryoprotective agent, preservation of isolated strains was assayed with several cryoprotective solutions. Methanol and glycerol were not suitable as cryoprotective agents for most of the isolates. Dimethyl sulfoxide (DMSO) was apparently the best cryoprotector. A new method, which used a filter paper as a growing substratum that later could be directly stored at −80°C, was successfully used. A morphological study of each strain under light and electron microscopy was made to classify them. All isolated strains belong to phylum BX, Class 1, subsection III of the Bergey's manual of systematic bacteriology, 2nd ed., vol. 1. Most genera are included in the LPP group as Lyngbya aestuarii and Microcoleus chthonoplastes. Received: 30 April 2002 / Accepted: 31 May 2002     

Effect of desert soil algae on the stabilization of fine sands

Four filamentous cyanobacteria, Microcoleusvaginatus, Phormidium tenue,Scytonemajavanicum (Kutz.) and Nostoc sp., and asingle-celled green alga, Desmococcus olivaceus, allisolated from Shapotou (Ningxia Hui Autonomous Region of China), were batchcultured and inoculated onto unconsolidated sand in greenhouse and fieldexperiments. Their ability to reduce wind erosion in sands was quantified byusing a wind tunnel laboratory. The major factors related to cohesion of algalcrusts, such as biomass, species, species combinations, bioactivity, niche,growth phase of algae, moisture, thickness of the crusts, dust accretion(including dust content and manner of dust added) and other cryptogams(lichens,fungi and mosses) were studied. The best of the five species were M.vaginatus and P. tenue, while the best mix wasablend of 80% M. vaginatus and 5% each of P.tenue,S. javanicum,Nostocsp. and D. olivaceus. The threshold friction velocity wassignificantly increased by the presence of all of the cyanobacterial species,while the threshold impact velocity was notably increased only by thefilamentous species. Thick crusts were less easily eroded than thin crusts,while biomass was more effective than thickness. Dust was incorporated bestintoMicrocoleus crust when added in small amounts over time,and appeared to increase growth of the cyanobacterium as well as strengthen thecohesion of the crust. Microbial crust cohesion was mainly attributed to algalaggregation, while lichens, fungi and mosses affected more the soil structureand physico-chemical properties.     

Trebouxia aggregata undGloeocapsa sanguinea,Phycobionten inEuopsis granatina (Lichinaceae)

The green algaTrebouxia aggregata (Archibald)Gärtner (Chlorellales) and the procaryotic blue-green alga (cyanobacterium)Gloeocapsa sanguinea (C. Agardh)Kütz. emend.Jaag (Chroococcales) are reported to be the phycobionts inEuopsis granatina (Sommerf.)Nyl. (Lichinaceae). The ultrastructure of the two organisms, studied in the lichen thallus, is described.

Frau Prof. Dr.Elisabeth Tschermak-Woess zu ihrem 70. Geburtstag gewidmet.     

Characteristics and role of the exocellular polysaccharides produced by five cyanobacteria isolated from phototrophic biofilms growing on stone monuments

Three coccoid and two filamentous cyanobacterial strains were isolated from phototrophic biofilms exposed to intense solar radiation on lithic surfaces of the Parasurameswar Temple and Khandagiri caves, located in Orissa State, India. Based on to their morphological features, the three coccoid strains were assigned to the genera Gloeocapsosis and Gloeocapsa, while the two filamentous strains were assigned to the genera Leptolyngbya and Plectonema. Eleven to 12 neutral and acidic sugars were detected in the slime secreted by the five strains. The secretions showed a high affinity for bivalent metal cations, suggesting their ability to actively contribute to weakening the mineral substrata. The secretion of protective pigments in the polysaccharide layers, namely mycosporine amino acid-like substances (MAAs) and scytonemins, under exposure to UV radiation showed how the acclimation response contributes to the persistence of cyanobacteria on exposed lithoid surfaces in tropical areas.     

Bioactive natural products from blue-green algae

Since 1981 we have cultured and prepared lipophilic and hydrophilic extracts from more than 1500 strains representing some 400 species of blue-green algae. Screening for a wide variety of potentially useful bioactivities, including cytotoxic, multi-drug-resistance reversal, antifungal, and antiviral effects, has led to the discovery and identification of numerous novel bioactive metabolites including peptides, macrolides and glycosides.A systematic evaluation of the chemical and environmental factors that influence the production of secondary metabolites inScytonema ocellatum, which produces tolytoxin (a macrocyclic lactone that depolymerizes actinin vivo to disrupt cell division in eukaryotic organisms), has shown that cyanophytes can be manipulated in culture to improve growth and product yields.     

On the use of high‐throughput sequencing for the study of cyanobacterial diversity in Antarctic aquatic mats

The study of Antarctic cyanobacterial diversity has been mostly limited to morphological identification and traditional molecular techniques. High‐throughput sequencing (HTS) allows a much better understanding of microbial distribution in the environment, but its application is hampered by several methodological and analytical challenges. In this work, we explored the use of HTS as a tool for the study of cyanobacterial diversity in Antarctic aquatic mats. Our results highlight the importance of using artificial communities to validate the parameters of the bioinformatics procedure used to analyze natural communities, since pipeline‐dependent biases had a strong effect on the observed community structures. Analysis of microbial mats from five Antarctic lakes and an aquatic biofilm from the Sub‐Antarctic showed that HTS is a valuable tool for the assessment of cyanobacterial diversity. The majority of the operational taxonomic units retrieved were related to filamentous taxa such as Leptolyngbya and Phormidium, which are common genera in Antarctic lacustrine microbial mats. However, other phylotypes related to different taxa such as Geitlerinema, Pseudanabaena, Synechococcus, Chamaesiphon, Calothrix, and Coleodesmium were also found. Results revealed a much higher diversity than what had been reported using traditional methods and also highlighted remarkable differences between the cyanobacterial communities of the studied lakes. The aquatic biofilm from the Sub‐Antarctic had a distinct cyanobacterial community from the Antarctic lakes, which in turn displayed a salinity‐dependent community structure at the phylotype level.     

Ultrastructure d'une cyanophycée aérienne calcifiée cavernicole:Scytonema julianum (Frank) Richter (Hormogonophycideae,Nostocales, Scytonemataceae)

The ultrastructure of the atmophytic lime-encrusted filamentous blue-green algaScytonema julianum (Frank) Richter which grows in caves, is studied here for the first time. The cytology of this organism is compared with that of another filamentous and lime-encrusted cyanophycea,Geitleria calcarea Friedmann, which grows in the same biotope. The authors also discuss the nomenclature of this quite rare species.

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Ultrastructure d'une cyanophycée aérienne calcifiée cavernicole:Scytonema julianum (Frank) Richter (Hormogonophycideae, Nostocales, Scytonemataceae)

     

A phylogenetic approach to species delimitation in freshwater Gastrotricha from Sweden

Gastrotricha is a cosmopolitan group of aquatic invertebrates. To date, approximately 765 species have been described. This study is the first to deal with species delimitation and cryptic species of freshwater Gastrotricha. Three commonly encountered species, Heterolepidoderma ocellatum, Lepidochaetus zelinkai, and Lepidodermella squamata, are investigated for cryptic speciation. Most of the material is based on Swedish specimens but closely related species from other parts of the world are also included. Taxonomic revisions are supported by phylogenies based on 18S rRNA, 28S rRNA, and COI mtDNA of freshwater Chaetonotidae from several genera and inferred from Bayesian and maximum likelihood approaches. Heterolepidoderma ocellatum f. sphagnophilum is raised to species level, under the name H. acidophilum n. sp. Moreover, genetic data based on COI indicate large variation between two morphologically very similar groups of Lepidodermella squamata. The extent of cryptic speciation in L. zelinkai appears low. Based on the phylogenetic hypothesis presented in this article, the new species, Lepidodermella intermedia n. sp., from northern Sweden is also described. The phylogenetic hypothesis generated shows that Chaetonotidae is a non-monophyletic group.