Using diatoms and physical and chemical parameters to monitor cow-pasture impact in peat cores from mountain mires.

Peatlands play a crucial role in carbon (C) sequestration and biodiversity conservation. However, these environments are highly vulnerable, and Europe has lost >60 % of its peatland habitat in recent decades. Cattle grazing and trampling contribute to peatland degradation, which generally result in a shift from moss-dominated vegetation to vascular plants and in lower C sequestration rates. Overgrazing poses also a significant threat to habitat integrity and biodiversity, especially in the Alpine area, where close-to-pristine mires with high ecological integrity are becoming extremely rare. Thus, a more in depth understanding of how cattle grazing and trampling are threatening Alpine mires is strongly needed for a sustainable management and conservation of these habitats. The objective of this study was to examine the impact of grazing on the physical, chemical, and biological characteristics of peat, with a focus on diatoms. To answer such a question, seven 50-cm deep cores were collected from mires located in the Adamello-Brenta Nature Park (North of Italy) along a grazing-induced disturbance gradient. Results indicated that grazing primarily affected at least the upper 15 cm of the peat, resulting in increased density and reduced water content, due to compaction, and lower C-to­nitrogen ratio, possibly caused by both cow manure inputs and increased peat mineralization. Moreover, almost 200 diatom taxa were recorded across the 7 cores, with several of them falling under threat categories in the Red List for central Europe. The higher percentage of eutraphentic species in highly-grazed areas was related to the increase in nutrients caused by cattle manure. Finally, intense grazing increased the share of taxa that are more likely to survive in environments with unstable water availability (= aerial species). We showed that diatom data, supported by physical and chemical parameters, can be a refined tool to inform mire protection and rehabilitation.

Fellowship of the Spring: An initiative to document and protect the world's oases.

An 'oasis' signifies a refugium of safety, recovery, relaxation, fertility, and productivity in an inhospitable desert, a sweet spot in a barren landscape where life-giving water spills forth from the Earth. Remarkable mythological congruencies exist across dryland cultures worldwide where oases or 'arid-land springs' occur. In many places they also provide specialised habitats for an extraordinary array of endemic organisms. To inform their management, and maintain their integrity, it is essential to understand the hydrogeology of aquifers and springs. Gravity-fed vs artesian aquifers; actively recharged vs fossil aquifers, and sources of geothermal activity are important concepts presented here. There consequences for oases of sustainable and unsustainable groundwater extraction, and other examples of effective conservation management. Oases are archetypes for human consciousness, habitats that deserve protection and conservation, and a lingua franca for multicultural values and scientific exchange. We represent an international Fellowship of the Spring seeking to encompass and facilitate the stewardship of oases and aquifers through improved knowledge, outreach, and governance.

Integrative Taxonomic, Ecological and Genotyping Study of Charophyte Populations from the Egyptian Western-Desert Oases and Sinai Peninsula.

Present-day information available on the charophyte macroalgae in Egypt, including their phylogenetic affinities, remains largely incomplete. In this study, nine charophyte populations were collected from different aquatic biotopes across the Egyptian Western-Desert Oases and Sinai Peninsula. All populations were investigated using an integrative polyphasic approach including phylogenetic analyses inferred from the chloroplast-encoded gene (rbcL) and the internal transcribed spacer (ITS1) regions, in parallel with morphotaxonomic assignment, ultrastructure of the oospore walls, and autecology. The specimens identified belonged to the genera Chara, Nitella, and Tolypella, with predominance of the first genus to which five species were assigned though they presented some interesting aberrant taxonomic features: C. aspera, C. contraria, C. globata, C. tomentosa, and C. vulgaris. Based on our integrative study, the globally rare species C. globata was reported for the second time for the whole African continent. The genus Nitella was only represented by N. flagellifera, and based on the available literature, it is a new record for North Africa. Noteworthy, an interesting Tolypella sp., morphologically very similar to T. glomerata, was collected and characterized and finally designated with the working name 'Tolypella sp. PBA-1704 from a desert, freshwater wetland', mainly based on its concatenated rbcL+ITS1 phylogenetic position. This study not only improved our understanding on the diversity, biogeography and autecological preferences of charophytes in Egypt, but it also broadened our knowledge on this vulnerable algal group in North Africa, emphasizing the need of more in-depth research work in the future, particularly in the less-impacted desert habitats.

Geological and hydrochemical prerequisites of unexpectedly high biodiversity in spring ecosystems at the landscape level.

This study explores the factors affecting the biodiversity of diatoms, vegetation with focus on bryophytes, and invertebrates with focus on water mites, in a series of 16 spring-habitats. The springs are located primarily from the mountainous part of the Emilia-Romagna Region (Northern Apennines, Italy), and two pool-springs from agricultural and industrial lowland locations. Overall, data indicate that biological diversity (Shannon-Wiener, α-diversity) within individual springs was relatively low, e.g.: Sdiatoms = 0-46, Swater-mites = 0-11. However, when examined at the regional scale, they hosted a very high total number of taxa (γ-diversity; Sdiatoms = 285, Swater-mites = 40), including several new or putatively-new species, and many Red-List taxa. This pattern suggested there is high species turnover among springs, as well as high distinctiveness of individual spring systems. A key goal was to assess the hydrogeological and hydrochemical conditions associated with this high regional-pool species richness, and to provide a guide to future conservation strategies. There was a striking variety of geological conditions (geodiversity, captured mainly with lithotype and aquifer structure) across the study region, which led to wide variation in the hydrosphere, especially in conductivity and pH. Agriculture and industrial activities (anthroposphere) in the lowlands resulted in nutrient enrichment and other forms of pollution. Across all 16 spring-systems, several hydrogeological conditions most strongly influenced the presence or absence of particular biota and were determinants of species importance: spring-head morphology, hydroperiod, discharge, current velocity, and elemental concentration. These findings have important practical consequences for conservation strategies. Our data show that it is imperative to protect entire regional groups of springs, including representatives of the different ecomorphological spring types, lithologies, and degrees of human influence. These findings suggest that springs, when studied from an ecohydrogeological perspective, are excellent systems in which to further investigate and understand geo-biodiversity relationships.

Novel green algal isolates from the Egyptian hyper-arid desert oases: a polyphasic approach with a description of Pharao desertorum gen. et sp. nov. (Chlorophyceae, Chlorophyta).

The biodiversity of terrestrial algae is still grossly understudied, and African deserts in particular are barely touched in this respect. Here, four coccoid green algae from oases in the Western Desert of Egypt were characterized using a combination of morphotaxonomic, ecological and 18S rDNA data, with additional carotenoid and lipid analyses for two of the strains. Three strains were identified as affiliated with known taxa: Mychonastes sp., Asterarcys sp. (first report of this genus from a desert soil), and Stichococcus cf. deasonii. The fourth strain is proposed to represent a new cryptic genus Pharao gen. nov., with the type species P. desertorum sp. nov. The new taxon is sister to the clade of uncharacterized North American desert strains of Radiococcaceae (Chlorophyceae, Chlorophyta). The pigment profile of P. desertorum gen. et sp. nov. revealed carotenoids and chlorophylls typical of green algae. Bioorganic analysis showed a complex lipidome based on phospho- (PC), galacto- (MGDG and DGDG), betaine- (DGTS), and sulfoquinovosyl- (SQDG) membrane lipids, besides significant amounts of storage neutral lipids such as diacyl- (DAG) and triacylglycerols (TAG). The presence of saturated alkyl chains within all the membrane lipid classes in P. desertorum and Asterarcys sp. appears to reflect the need to maintain membrane fluidity and viscosity. In summary, African deserts likely still harbor new taxa to be described, and lipidomic analyses of such taxa may provide clues about their ability to survive in the extremely harsh desert habitats.

A global review on ambient Limestone-Precipitating Springs (LPS): Hydrogeological setting, ecology, and conservation.

Springs are biodiversity hotspots and unique habitats that are threatened, especially by water overdraft. Here we review knowledge on ambient-temperature (non-geothermal) freshwater springs that achieve sufficient oversaturation for CaCO3 -by physical CO2 degassing and activity of photoautotrophs- to deposit limestone, locally resulting in scenic carbonate structures: Limestone-Precipitating Springs (LPS). The most characteristic organisms in these springs are those that contribute to carbonate precipitation, e.g.: the mosses Palustriella and Eucladium, the crenophilous desmid Oocardium stratum, and cyanobacteria (e.g., Rivularia). These organisms appear to be sensitive to phosphorus pollution. Invertebrate diversity is modest, and highest in pools with an aquatic-terrestrial interface. Internationally, comprehensive legislation for spring protection is still relatively scarce. Where available, it covers all spring types. The situation in Europe is peculiar: the only widespread spring type included in the EU Habitat Directive is LPS, mainly because of landscape aesthetics. To support LPS inventorying and management to meet conservation-legislation requirements we developed a general conceptual model to predict where LPS are more likely to occur. The model is based on the pre-requisites for LPS: an aquifer lithology that enables build-up of high bicarbonate and Ca(2+) to sustain CaCO3 oversaturation after spring emergence, combined with intense groundwater percolation especially along structural discontinuities (e.g., fault zones, joints, schistosity), and a proper hydrogeological structure of the discharging area. We validated this model by means of the LPS information system for the Emilia-Romagna Region (northern Italy). The main threats to LPS are water diversion, nutrient enrichment, and lack of awareness by non-specialized persons and administrators. We discuss an emblematic case study to provide management suggestions. The present review is devoted to LPS but the output of intense ecological research in Central Europe during the past decades has clearly shown that effective conservation legislation should be urgently extended to comprise all types of spring habitats.

Benthic algal assessment of ecological status in European lakes and rivers: Challenges and opportunities.

This opinion paper introduces a special series of articles dedicated to freshwater benthic algae and their use in assessment and monitoring. This special series was inspired by talks presented at the 9th International Congress on the Use of Algae for Monitoring Rivers and Comparable Habitats (Trento, Italy, 2015), the latest of a series of meetings started in 1991. In this paper, we will first provide a brief overview of phytobenthos methods in Europe. Then, we will turn towards the 'dark side' of phytobenthos and describe four particular problems for phytobenthos assessment in the European Union: (1) over-reliance on a single group of algae (mostly diatoms) to the exclusion of other groups; (2) relatively low adoption of benthic algae for ecological assessments in lakes; (3) absence of measures of phytobenthos abundance; (4) approaches used to define boundaries between ecological classes. Following this, we evaluate the strengths and limitations of current phytobenthos assessment methods against 12 criteria for method evaluation addressing four areas: ecological rationale, performance, feasibility of implementation, and use in communication and management. Using these criteria, we identify and discuss three general challenges for those developing new methods for phytobenthos-based assessment: a weak ecological rationale and insufficient consideration of the role of phytobenthos as a diagnostic tool and for communicating ecosystem health beyond a narrow group of specialists. The papers in the special series allow a comparison with the situation and approaches in the USA, present new methods for the assessment of ecological status and acidification, provide tools for an improved management of headwaters and petrifying springs, discuss the utility of phytobenthos for lake assessments, and test the utility of functional measures (such as biofilm phosphorus uptake capacity, PUC).

Achnanthidium minutissimum (Bacillariophyta) valve deformities as indicators of metal enrichment in diverse widely-distributed freshwater habitats.

In the presence of different environmental stressors, diatoms can produce frustules presenting different types of deformities. Metals and trace elements are among the most common causes of these teratological forms. Metal enrichment in water bodies can be attributed to the geological setting of the area or to pollution. The widespread benthic diatom Achnanthidium minutissimum (ADMI) is one of the most metal-tolerant species. In the present study, ADMI teratologies were defined from samples taken from eight very diverse, widely-distributed inland-water habitats: streams affected by active and abandoned mining areas, a metal-contaminated stream, a spring in an old chalcopyrite mine, a mineral-water fountain, and a sediment core taken from a lake affected by metal contamination in the past. Deformed frustules of ADMI were characterised mainly by one (sometimes two) more or less bent off ending, conferring to the specimens a cymbelloid outline (cymbelliclinum-like teratology, CLT). Marked teratologies were distinguished from slight deformities. Hydrochemical analyses, including metals and trace elements, were carried out and enrichment factors (EF) relative to average crustal composition were calculated. To improve our knowledge on the potential of different metals and trace elements to trigger the occurrence of ADMI CLT, we carefully selected 15 springs out of 110 (CRENODAT dataset) where both ADMI and above-average metal or metalloid concentrations occurred, and re-analysed these samples. The results from the eight widely-distributed core sites as well as from the 15 selected CRENODAT springs led to the hypothesis that two metals (copper and zinc) and a metalloid (antimony) were the most likely triggers of ADMI CLT formation. From a quantitative point of view, it is worth noting that the lowest concentrations triggering ADMI CLT can be fairly low, particularly in the case of copper contamination. The antimony-rich site was characterised by a marked-teratology variant where both ends of ADMI were bent off.

Fine-Scale Spatial Patterns of Mature and Immature Filaments and Bioorganic Compounds of the Rhodophyte Bangia atropurpurea in the Supra- and Eulittoral Zones of a Large Lake(1).

Bangia atropurpurea (Mertens ex Roth) C. Agardh is a freshwater red alga species that is distributed worldwide. B. atropurpurea is highly adaptable due to its stress-tolerance, which ensures survival under desiccation periods and under radiation extremes typical of the supra- and upper eulittoral zones. Whereas a number of previous investigations addressed some of the physiological and biochemical traits involved in stress-tolerance, we studied the spatial arrangement of the mature (multiseriate) and immature (uniseriate) filaments and of selected bioorganic compounds along a gradient defined by distance from the waterline. Substantial physiological and biochemical differences were previously observed among phenological stages in the marine environment. In this study, we showed a nonrandom spatial structure of both phenological stages and photosynthetic pigments and photoprotective compounds, R-phycocyanin and R-phycoerythrin along the supralittoral-eulittoral gradient. This observed pattern strongly suggests a complex interplay between physio-morphological regulation and spatial arrangement of mature and immature filaments in conferring the typical stress tolerance of B. atropurpurea.

MICROFISSURATA GEN. NOV. (BACILLARIOPHYTA), A NEW DIATOM GENUS FROM DYSTROPHIC AND INTERMITTENTLY WET TERRESTRIAL HABITATS(1).

A new freshwater benthic diatom genus, Microfissurata gen. nov., was identified from light and scanning electron micrographs. The most characteristic morphological feature (referred to also by the name of the new genus) is the structure of the striae/alveoli, which are simple, mostly uninterrupted, transapical slits. The combination of characteristics of the new genus is unique. It includes two new species: Microfissurata paludosa Cantonati et Lange-Bert. sp. nov. and M. australis Van de Vijver et Lange-Bert. sp. nov., distributed in Europe and on an austral island, respectively. The new genus occurs in freshwater dystrophic lakes, pools, seepage springs, mires, and in intermittently wet terrestrial habitats. It is not abundant but widespread wherever these habitat types are well developed, in Nordic-alpine and sub-Antarctic areas. Overall, the new genus appears to be mostly epiphytic (bryophilous) and capable of tolerating a wide range of moisture conditions (xerotolerant). The finding of the type species (M. paludosa) in a well-investigated area like central Europe highlights the diatom species richness of dystrophic habitats and their importance for diatom biodiversity conservation.

A new method for the identification and the structural characterisation of carotenoid esters in freshwater microorganisms by liquid chromatography/electrospray ionisation tandem mass spectrometry.

Liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS) has been employed to identify carotenoid esters present in raw organic extracts of pigmented freshwater microalgae and to gain structural information on these compounds. In particular, acyl carotenoid derivatives of Haematococcus pluvialis and Euglena sanguinea have been characterised by tandem mass spectrometry (MS/MS) in a quadrupole ion trap. ESI-MS/MS allows recognition of the presence of carotenoid esters in complicated mixtures without any initial chromatographic work-up and without the need to use UV-Vis photo-diode array (PDA) detectors. Product ion scans of the [M + Na]+ ion lead to known neutral losses of the C7H8 and C8H10 residues from the conjugated polyene moiety of the carotenoid unit, that permit the unambiguous identification of the carotenoid itself. These structurally relevant ions are not observed in positive or negative ion APCI (atmospheric pressure chemical ionisation) mass spectra. Moreover, the several product ions observed in positive and/or negative ion ESI-MS/MS not only are a diagnostic signature of the main structural features of the acyl chains such as length, position and unsaturation, but also display the nominal mass of the parent xanthophyll. Our methodology has been validated (i) by using esters of astaxanthin obtained from off-line purification of the H. pluvialis extracts and structurally elucidated through proton nuclear magnetic resonance (1H-NMR) spectroscopy and (ii) by product analysis of esters by alkaline hydrolysis. The characterisation of the unknown carotenoid esters of E. sanguinea is a demonstration of the capabilities of this methodology.

On-line identification of secondary metabolites in freshwater microalgae and cyanobacteria by combined liquid chromatography-photodiode array detection-mass spectrometric techniques.

The analysis and identification of a wide range of secondary metabolites biosynthesized by different algal taxa and cyanobacteria has been performed through a selective and sensitive methodology, mainly based on reversed-phase HPLC coupled both to UV photodiode array detection and to atmospheric pressure mass spectrometric techniques (HPLC-DAD-APIMS). Results are reported here with special attention to the analyses carried out both on the natural phytoplankton (mixed populations) of Lake Tovel (Northern Italy, Brenta Dolomites) and on enclosure-produced biomass of the dinoflagellate Glenodinium sanguineum Marchesoni (1941). This analytical procedure might represent a powerful tool for the fast screening of the taxonomic composition (broad groups, e.g. divisions) of natural mixed populations of phytoplankton, by providing a reliable distribution of accessory pigments extracted from microalgae, such as carotenoids and chlorophyll derivatives. Furthermore, we showed that in the same chromatographic analysis other classes of natural products, such as galactolipids, alkaloids, sterols and mycosporine-like amino acids, can be detected by using combined optical and mass spectrometric techniques. These metabolites represent distinctive biochemical signatures, sometimes even at the species level.