Assessment of in-situ monitoring and tracking the vertical migration of cyanobacterial blooms using LISST-HAB.

Cyanobacterial harmful algal blooms (cyanoHABs) are becoming increasingly common in aquatic ecosystems worldwide. However, their heterogeneous distributions make it difficult to accurately estimate the total algae biomass and forecast the occurrence of surface cyanoHABs by using traditional monitoring methods. Although various optical instruments and remote sensing methods have been employed to monitor the dynamics of cyanoHABs at the water surface (i.e., bloom area, chlorophyll a), there is no effective in-situ methodology to monitor the dynamic change of cell density and integrated biovolume of algae throughout the water column. In this study, we propose a quantitative protocol for simultaneously measurements of multiple indicators (i.e., biovolume concentration, size distribution, cell density, and column-integrated biovolume) of cyanoHABs in water bodies by using the laser in-situ scattering and transmissometry (LISST) instrument. The accuracy of measurements of the biovolume and colony size of algae was evaluated and exceeded 95% when the water bloom was dominated by cyanobacteria. Furthermore, the cell density of cyanobacteria was well estimated based on total biovolume and mean cell volume measured by the instrument. Therefore, this methodology has the potential to be used for broader applications, not only to monitor the spatial and temporal distribution of algal biovolume concentration but also monitor the vertical distribution of cell density, biomass and their relationship with size distribution patterns. This provides new technical means for the monitoring and analysis of algae migration and early warning of the formation of cyanoHABs in lakes and reservoirs.

Nuclear genome of dinoflagellates: Size variation and insights into evolutionary mechanisms.

Recent progress in high-throughput sequencing technologies has dramatically increased availability of genome data for prokaryotes and eukaryotes. Dinoflagellates have distinct chromosomes and a huge genome size, which make their genomic analysis complicated. Here, we reviewed the nuclear genomes of core dinoflagellates, focusing on the genome and cell size. Till now, the genome sizes of several dinoflagellates (more than 25) have been measured by certain methods (e.g., flow cytometry), showing a range of 3-250 pg of genomic DNA per cell. In contrast to their relatively small cell size, their genomes are huge (about 1-80 times the human haploid genome). In the present study, we collected the genome and cell size data of dinoflagellates and compared their relationships. We found that dinoflagellate genome size exhibits a positive correlation with cell size. On the other hand, we recognized that the genome size is not correlated with phylogenetic relatedness. These may be caused by genome duplication, increased gene copy number, repetitive non-coding DNA, transposon expansion, horizontal gene transfer, organelle-to-nucleus gene transfer, and/or mRNA reintegration into the genome. Ultimate verification of these factors as potential causative mechanisms would require sequencing of more dinoflagellate genomes in the future.

Summer monsoon promotes the energy transfer efficiency of the zooplankton community in northern South China sea.

The summer monsoon shows a fundamental influence on the pelagic ecosystem of the South China Sea. Zooplankton are a major link for energy transfer between primary producers and upper trophic levels. Therefore, evaluating the energy transfer efficiency (ETE) of zooplankton is crucial to understand the function of pelagic ecosystem under the influence of monsoon. In this study, field surveys were conducted during May (intermonsoon) and August 2021 (summer monsoon) focusing on the variation of zooplankton size and trophic structures across the shelf and slope. The result showed that the summer monsoon reinforced the gradient of abundance, biovolume, and biomass from slope to shelf, and greatly intensified the role of environmental factors in driving spatial variation in most taxa. Both the results of size and trophic structures indicated that the ETE of zooplankton decreased from slope to shelf. The size structure also indicated that the ETE of zooplankton significantly increased under the influence of summer monsoon. These results were consistent with previous studies by different methods, suggesting that these approaches of size and trophic structures had important potential value in assessing changes in the function of marine pelagic ecosystem, especially when compared with sufficient historical data or reanalyzing historical samples.

Dataset on meteorological forcing mechanisms impacting marine circulation and oceanographic variables in the northern part of the Veracruz reef system.

The dataset is divided in two main groups. The first group, referred to as "Meteorological data", consists of air temperature, sea level pressure and U and V components of wind direction and intensity: The second group, referred to as "Oceanographic data", includes biovolume, sea level and water temperature measurements. The meteorological data is derived from model data obtained from the NAAR-NCEP reanalysis for North America, calculated over the area of the Veracruz reef system, Mexico. On the other hand, the oceanographic data was collected in situ using four ADCPs (Acoustic Doppler Current Profilers) anchored at a depth of 20 m at four different reefs within the Veracruz reef system. Both datasets cover a period of 10 days in November 2008, during which successive low-and high-pressure systems occurred over the southwestern Gulf of Mexico. These datasets can be used to evaluate the effect of the pressure changes on marine circulation, residual current and oceanographic variables.

Estimation of cellular carbon content based on the cell biovolume of microalgae from a eutrophic estuary (Sea of Marmara, Türkiye).

The cellular carbon content based on the cell biovolume of a total of 61 microalgal species determined in a eutrophic estuary (Golden Horn, Sea of Marmara) was estimated in seawater samples taken during two different sampling periods. Cell biovolume according to geometric dimensions of the cells was then converted to phytoplankton carbon using an appropriate conversion factor. The range of diatom biovolume, in which the majority had small cell sizes (<50 µm), was much wider than that of dinoflagellate biovolume, in which the majority had large cell sizes (>50 µm). The cell biovolume and carbon content ranged from 35 to 4.88 × 105 µm3 and 5 to 1.18 × 104 pgC cell-1 for diatoms and from 3.66 × 102 to 8.68 × 105 µm3 and 55 to 8.14 × 104 pgC cell-1 for dinoflagellates, respectively. The mean carbon density for diatoms and dinoflagellates (excl. Noctiluca scintillans) varied between 0.027 and 0.099 pgC µm-3 0.096 and 0.136 pgC µm-3, respectively. The mean cell carbon content and carbon density of dinoflagellates (6.73 × 103 pgC cell-1 and 0.115 pgC µm-3) were approximately 10 and 2 times greater than those of diatoms, respectively. The carbon content of the other phytoflagellates was lower, whereas their carbon density was higher. As a result, the findings from this study will provide a significant contribution to the assessment and estimation of carbon biomass during algal blooms in this study area.

Uncertainties of cell number estimation in cyanobacterial colonies and the potential use of sphere packing.

Cyanobacteria are notorious bloom formers causing various water quality concerns, such as toxin production, extreme diurnal variation of oxygen, pH, etc., therefore, their monitoring is essential to protect the ecological status of aquatic systems. Cyanobacterial cell counts and biovolumes are currently being used in water management and water quality alert systems. In this study, we investigated the accuracy of traditional colonial biovolume and cell count estimation approaches used in everyday practice. Using shape realistic 3D images of cyanobacterial colonies, we demonstrated that their shape cannot be approximated by ellipsoids. We also showed that despite the significant relationship between overall colony volume and cell biovolumes, because of the considerable scatter of cell count data the regressions give biased estimates for cyanobacterial cell counts. We proposed a novel approach to estimate cell counts in colonies that was based on the random close sphere packing method. This method provided good results only in those cases when overall colony volumes could be accurately measured. The visual investigation of colonies done by skilled experts has given precise but lower estimates for cell counts. The estimation results of several experts were surprisingly good, which suggests that this capability can be improved and estimation bias can be reduced to the level acceptable for water quality estimations.

Imaging and plate counting to quantify the effect of an antimicrobial: A case study of a photo-activated chlorine dioxide treatment.

AIM: To assess removal versus kill efficacies of antimicrobial treatments against thick biofilms with statistical confidence. METHODS AND RESULTS: A photo-activated chlorine dioxide treatment (Photo ClO2 ) was tested in two independent experiments against thick (>100 µm) Pseudomonas aeruginosa biofilms. Kill efficacy was assessed by viable plate counts. Removal efficacy was assessed by 3D confocal scanning laser microscope imaging (CSLM). Biovolumes were calculated using an image analysis approach that models the penetration limitation of the laser into thick biofilms using Beer's Law. Error bars are provided that account for the spatial correlation of the biofilm's surface. The responsiveness of the biovolumes and plate counts to the increasing contact time of Photo ClO2 were quite different, with a massive 7 log reduction in viable cells (95% confidence interval [CI]: 6.2, 7.9) but a more moderate 73% reduction in biovolume (95% CI: [60%, 100%]). Results are leveraged to quantitatively assess candidate CSLM experimental designs of thick biofilms. CONCLUSIONS: Photo ClO2 kills biofilm bacteria but only partially removes the biofilm from the surface. To maximize statistical confidence in assessing removal, imaging experiments should use fewer pixels in each z-slice, and more importantly, at least two independent experiments even if there is only a single field of view in each experiment. SIGNIFICANCE AND IMPACT OF STUDY: There is limited penetration depth when collecting 3D confocal images of thick biofilms. Removal can be assessed by optimally fitting Beer's Law to all of the intensities in a 3D image and by accounting for the spatial correlation of the biofilm's surface. For thick biofilms, other image analysis approaches are biased or do not provide error bars. We generate unbiased estimates of removal and assess candidate CSLM experimental designs of thick biofilms with different pixilations, numbers of fields of view and number of experiments using the included design tool.

Changes in Phytoplankton Community Composition and Phytoplankton Cell Size in Response to Nitrogen Availability Depend on Temperature.

The climate-driven changes in temperature, in combination with high inputs of nutrients through anthropogenic activities, significantly affect phytoplankton communities in shallow lakes. This study aimed to assess the effect of nutrients on the community composition, size distribution, and diversity of phytoplankton at three contrasting temperature regimes in phosphorus (P)-enriched mesocosms and with different nitrogen (N) availability imitating eutrophic environments. We applied imaging flow cytometry (IFC) to evaluate complex phytoplankton communities changes, particularly size of planktonic cells, biomass, and phytoplankton composition. We found that N enrichment led to the shift in the dominance from the bloom-forming cyanobacteria to the mixed-type blooming by cyanobacteria and green algae. Moreover, the N enrichment stimulated phytoplankton size increase in the high-temperature regime and led to phytoplankton size decrease in lower temperatures. A combination of high temperature and N enrichment resulted in the lowest phytoplankton diversity. Together these findings demonstrate that the net effect of N and P pollution on phytoplankton communities depends on the temperature conditions. These implications are important for forecasting future climate change impacts on the world's shallow lake ecosystems.

The impact of micropollutants on native algae and cyanobacteria communities in ecological filters during drinking water treatment.

An attractive alternative for drinking water production is ecological filtration. Previous studies have reported high removal levels of pharmaceutical and personal care products (PPCPs) by this technology. Algae and cyanobacteria play an important role in the biological activity of ecological filters. The aim of this study was to characterize and identify the community of algae and cyanobacteria in relation to its composition, density and biovolume from 22 ecological filters that received spikings of 2 µg L-1 PPCPs. For algae and cyanobacteria species, triplicate samples were collected before and 96 h after each spiking from the interface between the top sand layer of the ecological filters and the supernatant water. Results show that Chlorophyceae and Cyanobacteria were present in high numbers of taxa and abundance. The specie Lepocinclis cf. ovum (Euglenophyceae) had the highest percentage occurrence/abundance and frequency into the filters, indicating a possible tolerance by Lepocinclis cf. ovum to the concentration of selected PPCPs. Although the concentration of PPCPs did not affect the treated water quality, they did affect the algae and cyanobacteria community. No differences were detected between filters that received a single PPCP and filters that received a mixture of the six compounds. Also, changes in the composition of algae and cyanobacteria communities were observed before and 96 h after the spikings.

Effects of space and environment on phytoplankton distribution in subtropical reservoirs depend on functional features of the species.

Environmental factors and dispersal can influence the structure of biological communities. Their effects can depend on the functional features of the species in the community. Since species belonging to the same trophic level, such as phytoplankton, may show functional differences, we investigated whether the effects of environment and dispersal differ among phytoplankton species from different functional groups. We analyzed data from a rainy and a dry period in 30 reservoirs in a subtropical region. In both periods, the environment as well as high and limited dispersal influenced the metacommunity structure. The functional groups had a low correspondence in their response to both dispersal and environment. Our results showed that the influence of the processes underlying the structure of the metacommunities, such as species sorting (environment influence), mass effect (high dispersal), and neutral dynamics (limited dispersal), depended on the functional characteristics of the organisms and could vary even among species of the same trophic level. These findings suggested that species at the same trophic level could not be considered ecological equivalents. This paper includes a Portuguese and Spanish version of the abstract in the online resources. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00027-021-00837-0.

Activity density at a continental scale: What drives invertebrate biomass moving across the soil surface?

Activity density (AD), the rate that an individual taxon or its biomass moves through the environment, is used both to monitor communities and quantify the potential for ecosystem work. The Abundance Velocity Hypothesis posited that AD increases with aboveground net primary productivity (ANPP) and is a unimodal function of temperature. Here we show that, at continental extents, increasing ANPP may have nonlinear effects on AD: increasing abundance, but decreasing velocity as accumulating vegetation interferes with movement. We use 5 yr of data from the NEON invertebrate pitfall trap arrays including 43 locations and four habitat types for a total of 77 habitat-site combinations to evaluate continental drivers of invertebrate AD. ANPP and temperature accounted for one-third to 92% of variation in AD. As predicted, AD was a unimodal function of temperature in forests and grasslands but increased linearly in open scrublands. ANPP yielded further nonlinear effects, generating unimodal AD curves in wetlands, and bimodal curves in forests. While all four habitats showed no AD trends over 5 yr of sampling, these nonlinearities suggest that trends in AD, often used to infer changes in insect abundance, will vary qualitatively across ecoregions.

Acoustic scattering by gas-bearing cyanobacterium Microcystis: Modeling and in situ biomass assessment.

Cyanobacterial harmful algal blooms (HABs) are increasing in a growing number of aquatic ecosystems around the world due to eutrophication and climatic change over the past few decades. Quantitative monitoring of HABs remains a challenge because their distributions are spatially heterogeneous and temporally variable. Most of the standard biological sampling methods are labor intensive and time consuming. In this paper, we present an efficient acoustic method to assess the biomass (biovolume) concentration of the cyanobacterium Microcystis in aquatic ecosystems. Acoustic backscattering vertical profiles from a gas-bearing Microcystis population were measured with echosounders at three frequencies (70, 120, and 333 kHz) in Lake Kinneret (case study). Concurrently, the volume concentration of Microcystis colonies and cyanobacteria-related Chlorophyll a were evaluated. We developed a partially coherent acoustic scattering model to quantify the cyanobacterium biomass based on depth-dependent acoustic backscattering signals. We also evaluated empirical regression models to obtain the Microcystis biomass from acoustically measured volume backscattering strength, Sv. It is demonstrated that both methods can convert the Sv to Microcystis biovolume concentrations reasonably well. Pro and cons of these methods are discussed. The results suggest that the presented methods may have a potential to be used for broader applications to monitor and quantify the gas-containing plankton in large aquatic ecosystems.

Biovolume and surface area calculations for microalgae, using realistic 3D models.

Morphology and spatial dimensions of microalgal units (cells or colonies) are among the most relevant traits of planktic algae, which have a pronounced impact on their basic functional properties, like access to nutrients or light, the velocity of sinking or tolerance to grazing. Although the shape of algae can be approximated by geometric forms and thus, their volume and surface area can be calculated, this approach cannot be validated and might have uncertainties especially in the case of complicated forms. In this study, we report on a novel approach that uses real-like 3D mesh objects to visualize microalgae and calculates their volume and surface area. Knowing these dimensions and their intraspecific variabilities, we calculated specific shape and surface area constants for more than 300 forms, covering more than two thousand taxa. Using these constants, the accurate volume and surface area can be quickly computed for each taxon and having these values, morphology-related metrics like surface area/volume ratio, the diameter of spherical equivalent can also be given quickly and accurately. Besides their practical importance, the volume and surface area constants can be considered as size-independent morphological traits that are characteristic for the microalgal shapes, and provide new possibilities of data analyses in the field of phytoplankton ecology.

Substantial intraspecific genome size variation in golden-brown algae and its phenotypic consequences.

BACKGROUND AND AIMS: While nuclear DNA content variation and its phenotypic consequences have been well described for animals, vascular plants and macroalgae, much less about this topic is known regarding unicellular algae and protists in general. The dearth of data is especially pronounced when it comes to intraspecific genome size variation. This study attempts to investigate the extent of intraspecific variability in genome size and its adaptive consequences in a microalgal species. METHODS: Propidium iodide flow cytometry was used to estimate the absolute genome size of 131 strains (isolates) of the golden-brown alga Synura petersenii (Chrysophyceae, Stramenopiles), identified by identical internal transcribed spacer (ITS) rDNA barcodes. Cell size, growth rate and genomic GC content were further assessed on a sub-set of strains. Geographic location of 67 sampling sites across the Northern hemisphere was used to extract climatic database data and to evaluate the ecogeographical distribution of genome size diversity. KEY RESULTS: Genome size ranged continuously from 0.97 to 2.02 pg of DNA across the investigated strains. The genome size was positively associated with cell size and negatively associated with growth rate. Bioclim variables were not correlated with genome size variation. No clear trends in the geographical distribution of strains of a particular genome size were detected, and strains of different genome size occasionally coexisted at the same locality. Genomic GC content was significantly associated only with genome size via a quadratic relationship. CONCLUSIONS: Genome size variability in S. petersenii was probably triggered by an evolutionary mechanism operating via gradual changes in genome size accompanied by changes in genomic GC content, such as, for example, proliferation of transposable elements. The variation was reflected in cell size and relative growth rate, possibly with adaptive consequences.

Reproductive cycle progression arrest and modification of cell morphology (shape and biovolume) in the alga Pseudokirchneriella subcapitata exposed to metolachlor.

Metolachlor (MET) is an herbicide widely used and frequently found (at µg L-1) in aquatic systems. This work aimed to study the modes of action of MET on the green microalga Pseudokirchneriella subcapitata. Algae exposed to 115 or 235 µg L-1 MET, for 48 or 72 h, presented a reduction of metabolic activity, chlorophyll a and b content and photosynthetic efficiency. The exposure to 115 or 235 µg L-1 MET also induced growth yield reduction, mean cell biovolume increase and alteration of the typical algae shape (cells lunate or helically twisted) to "French croissant"-type; at these MET concentrations, algal population was mainly composed by multinucleated cells (≥ 4 nuclei), which suggest that MET impairs the normal progression of the reproductive cycle but did not hinder nuclear division. The accumulation of multinucleated cells seems to be the consequence of the incapacity of the parent cell to release the autospores. In conclusion, MET disrupts the physiology of P. subcapitata cells; the disturbance of the progression of the reproductive cycle should be in the origin of growth slowdown (or even its arrest), increase of mean cell biovolume and modification of algal shape. This work contributed to elucidate, in a systematically and integrated way, the toxic mechanism of MET on the non-target organism, the alga P. subcapitata.

Características morfológicas de las cianobacterias y fitoplancton dominante en embalses de Antioquia: un enfoque basado en el biovolumen / Morphological characteristics of cyanobacteria and dominant phytoplankton in reservoirs in Antioquia: a focus based in biovolume

RESUMEN El desarrollo masivo de cianobacterias y la contaminación por cianotoxinas constituyen una problemática ambiental de especial interés en los sistemas acuáticos, debido a los diversos impactos negativos que los florecimientos (blooms) de cianobacterias pueden ocasionar. Dada la importancia recreacional y de abastecimiento de los embalses Abreo - Malpaso, Peñol - Guatapé y Playas, ubicados en el oriente de Antioquia - Colombia, en este estudio, se estimó la densidad de cianobacterias planctónicas y otros organismos del fitoplancton, así como el biovolumen geométrico, la relación superficie volumen (S/V) y la dimensión lineal máxima (DLM), a través de cuatro muestreos, en tres estaciones, para cada uno de los embalses. Se identificaron, dentro del phylum Cyanobacteria, con potencial de producción de cianotoxinas, el complejo Microcystis, los géneros Woronichinia, Aphanocapsa y Oscillatoria y la especie Radiocystis fernandoi. Al aplicar la prueba no parámetrica Kruskal - Wallis, no se encontraron diferencias significativas entre los embalses estudiados (p>0,05); sin embargo, el embalse El Peñol - Guatapé presentó la mayor biomasa algal y densidad de cianobacterias, por lo que, según los niveles de riesgo planteados para los embalses estudiados y de acuerdo con las características morfológicas de estos organismos, este embalse presenta un nivel de riesgo Medio Alto con relación a la presencia y morfología de las cianobacterias, mientras que los embalses Abreo - Malpaso y Playas están asociados a un riesgo Medio Bajo. Se sugiere establecer medidas de control y de prevención, con el fin de evitar un desarrollo masivo de cianobacterias, una potencial presencia de cianotoxinas y afectaciones graves a seres humanos y animales.

ABSTRACT Cyanotoxin contamination is of special interest in water bodies, especially those for human use, due to the various negative effects that blooms of potentially toxic cyanobacteria can cause on human health. Given the recreational and supply importance of the Abreo - Malpaso, Peñol - Guatapé and Playas reservoirs in the Department of Antioquia, Colombia. This study has estimated the biomass of the planktonic cyanobacteria, their geometric biovolume, the surface volume ratio (S / V) and the maximum linear dimension (MLD), during four sampling seasons in three different stations for each of these bodies. Water. We have identified from the Cyanophyta division with the production of cyanotoxins, the Microcystis complex and generates Woronichinia, Aphanocapsa and Oscillatoria, as well as the species Radiocystis fernandoi. When applying the nonparametric Kruskal - Wallis test, no significant differences were found between the studied reservoirs (p> 0.05); however, the Peñol - Guatapé reservoir presented the highest algae biomass and density of cyanobacteria, therefore, according to the levels of risk posed for the reservoirs studied and according to the morphological characteristics of these organisms, this reservoir presents a High Medium risk level in relation to the presence and morphology of the cyanobacteria, while the Abreo-Malpaso and Playas reservoirs are associated with a Medium Low risk. It is suggested to establish control and prevention measures to avoid a massive development of cyanobacteria, a possible presence of cyanotoxins and serious affectations in humans and animals.

Corrigendum: Can High Throughput Phenotyping Help Food Security in the Mediterranean Area?

[This corrects the article DOI: 10.3389/fpls.2019.00015.].

Can High Throughput Phenotyping Help Food Security in the Mediterranean Area?

According to the IPCC 2014 report the Mediterranean region will be affected by strong climatic changes, both in terms of average temperature and of precipitations regime. This area hosts some half a billion people and the impact on food production will be severe. To implement a climate smart agriculture paradigm and a sustainable increase of agricultural productivity different approaches can be deployed. Agriculture alone consumes 70% of the entire water available on the planet, thus the observed reduction of useful rainfall and growing costs for irrigation water may severely constrain food security. In our work we focused on two typical Mediterranean crops: durum wheat, a rainfed crop, and tomato, an irrigated one. In wheat we explored the possibility of identifying genotypes resilient to water stress for future breeding aims, while in tomato we explored the possibility of using biostimulants to increase the plant capacity of using water. In order to achieve these targets, we used high throughput phenotyping (HTP). Two traits were considered: digital biovolume, a measure based on imaging techniques in the RGB domain, and Water Use Efficiency index as calculated semi-automatically on the basis of evaporation measurements resulting in a high throughput, non-destructive, non-invasive approach, as opposed to destructive and time consuming traditional methods. Our results clearly indicate that HTP is able to discriminate genotypes and biostimulant treatments that allow plants to use soil water more efficiently. In addition, these methods based on RGB quality images can easily be scaled to field phenotyping structure USVs or UAVs.

Protist Diversity and Seasonal Dynamics in Skagerrak Plankton Communities as Revealed by Metabarcoding and Microscopy.

Protist community composition and seasonal dynamics are of major importance for the production of higher trophic levels, such as zooplankton and fish. Our aim was to reveal how the protist community in the Skagerrak changes through the seasons by combining high-throughput sequencing and microscopy of plankton collected monthly over two years. The V4 region of the 18S rRNA gene was amplified by eukaryote universal primers from the total RNA/cDNA. We found a strong seasonal variation in protist composition and proportional abundances, and a difference between two depths within the euphotic zone. Highest protist richness was found in late summer-early autumn, and lowest in winter. Temperature was the abiotic factor explaining most of the variation in diversity. Dinoflagellates was the most abundant and diverse group followed by ciliates and diatoms. We found about 70 new taxa recorded for the first time in the Skagerrak. The seasonal pattern in relative read abundance of major phytoplankton groups was well in accordance with microscopical biovolumes. This is the first metabarcoding study of the protist plankton community of all taxonomic groups and through seasons in the Skagerrak, which may serve as a baseline for future surveys to reveal effects of climate and environmental changes.

Cyanobacteria inhabiting biological soil crusts of a polar desert: Sør Rondane Mountains, Antarctica.

Molecular and morphological methods were applied to study cyanobacterial community composition in biological soil crusts (BSCs) from four areas (two nunataks and two ridges) in the Sør Rondane Mountains, Antarctica. The sampling sites serve as control areas for open top chambers (OTCs) that were put in place in 2010 at the time of sample collection and will be compared with BSC samples taken from the OTCs in the future. Cyanobacterial cell biovolume was estimated using epifluorescence microscopy, which revealed the dominance of filamentous cyanobacteria in all studied sites except the Utsteinen ridge, where unicellular cyanobacteria were the most abundant. Cyanobacterial diversity was studied by a combination of molecular fingerprinting methods based on the 16S rRNA gene (denaturing gradient gel electrophoresis (DGGE) and 454 pyrosequencing) using cyanobacteria-specific primers. The number of DGGE sequences obtained per site was variable and, therefore, a high-throughput method was subsequently employed to improve the diversity coverage. Consistent with previous surveys in Antarctica, both methods showed that filamentous cyanobacteria, such as Leptolyngbya sp., Phormidium sp. and Microcoleus sp., were dominant in the studied sites. In addition, the studied localities differed in substrate type, climatic conditions and soil parameters, which probably resulted in differences in cyanobacterial community composition. Furthermore, the BSC growing on gneiss pebbles had lower cyanobacterial abundances than BSCs associated with granitic substrates.