Environmental factors influencing the abundance and spatial distribution of modern dinoflagellate cysts in Izmir Bay (Eastern Mediterranean).

We analyzed surface sediments from 12 stations located in Izmir Bay to determine the impact of anthropogenic pollution on dinoflagellate cysts. Forty-three dinoflagellate cyst taxa and two cyst assemblage zones were identified. Zone 1 is characterized by the dominance of cysts of Gymnodinium nolleri, Selenopemphix nephroides, and Operculodinium centrocarpum from the oligotrophic part of Izmir Bay. Zone 2 is in the highly productive inner part of the bay and is characterized by the high abundances of Lingulodinium machaerophorum, Spiniferites ramosus, cysts of Scrippsiella spp., cysts of Polykrikos spp. and Quinquecuspis concreta. We used multivariate statistical analysis (DCA and CCA) on dinoflagellate cysts and environmental variables to support the identification of Zones 1 and 2. Our analyses also revealed that summer and winter chlorophyll-a as well as elevated nitrate and nitrite concentrations are significant parameters in controlling dinoflagellate cyst distribution in Izmir Bay.

Does dinocyst wall composition really reflect trophic affinity? New evidence from ATR micro-FTIR spectroscopy measurements.

Attenuated total reflection (ATR) microscope Fourier transform infrared (micro-FTIR) spectroscopy was used to investigate the dinosporin composition in the walls of modern, organic-walled dinoflagellate resting cysts (dinocysts). Variable cyst wall compositions were observed, which led to the erection of four spectrochemical groups, some with striking similarities to other resistant biomacromolecules such as sporopollenin and algaenan. Furthermore, possible proxies derivable from the spectrochemical composition of modern and fossil dinocysts were discussed. The color of the dinocyst walls was reflected in the spectral data. When comparing that color with a standard and the results of a series of bleaching experiments with oxidative agents, eumelanin was assigned as a likely pigment contributing to the observed color. Following this assignment, the role of eumelanin as an ultraviolet sunscreen in colored dinocysts was hypothesized, and its implications on the autofluorescence and morphological preservation of dinocysts were further discussed. Unlike what had previously been assumed, it was shown that micro-FTIR data from dinocysts cannot be used to unambiguously infer trophic affinities of their associated cells. Finally, using methods with high spatial resolutions (synchrotron transmission micro-FTIR and optical photothermal infrared spectroscopy), it was shown that dinocyst wall layers are chemically homogenous at the probed scales. This study fills a large knowledge gap in our understanding of the chemical nature of dinocyst walls and has nuanced certain assumptions and interpretations made in the past.

User and Professional Aspects for Sustainable Computing Based on the Internet of Things in Europe.

The commonly accepted definition of sustainability considers the availability of relevant resources to make an activity feasible and durable while also recognizing users' support as an essential part of the social side of sustainability. IoT represents a disruption in the general scenario of computing for both users and professionals. The real expansion and integration of applications based on IoT depend on our capacity of exploring the necessary skills and professional profiles that are essential for the implementation of IoT projects, but also on the perception of relevant aspects for users, e.g., privacy, legal, IPR, and security issues. Our participation in several EU-funded projects with a focus on this area has enabled the collection of information on both sides of IoT sustainability through surveys but also by collecting data from a variety of sources. Thanks to these varied and complementary sources of information, this article will explore the user and professional aspects of the sustainability of the Internet of Things in practice.

Salinity indicators in sediment through the fluvial-to-marine transition (Fraser River, Canada).

Many sediment attributes have been proposed as proxies for determining salinity conditions under which sediment is deposited, and six attributes (Sr/Ba-HAc, Sr/Ba-NH4Ac, δ13Corg, C/N, and the relative abundances and concentrations of dinoflagellate cysts) are compared here. In this paper, sediment attributes from the Fraser River Delta, Canada and surrounding coastal areas are compared by depositional position along the fluvial-to-marine transition, by salinity, and by sedimentological characteristics. Along the fluvial-to-marine transition, most attributes exhibit distinct trends between parts of the river that experience sustained marine water (saltwater) influence over seasonal and tidal timeframes, and parts that experience only freshwater or periodic saltwater influence. No attributes are reliable indicators of depositional position where saltwater incursion is short lived or where water is fresh. Where marine influence is sustained, Sr/Ba-HAc and Sr/Ba-NH4Ac are the most reliable positional indicators along the fluvial-to-marine transition. When compared strictly to salinity, Sr/Ba-HAc, Sr/Ba-NH4Ac, and δ13Corg all correlate predictably except in delta front and prodelta settings. Our data show that all six sediment attributes are heavily impacted by river-derived sedimentation, and it is not appropriate to compare values from strongly river-influenced settings (e.g., deltas) with those from weakly river-influenced settings (e.g., bays and estuaries).

Unraveling the Gonyaulax baltica Species Complex: Cyst-theca Relationship of Impagidinium variaseptum, Spiniferites pseudodelicatus sp. nov. and S. ristingensis (Gonyaulacaceae, Dinophyceae), With Descriptions of Gonyaulax bohaiensis sp. nov, G. amoyensis sp. nov. and G. portimonensis sp. nov.

The taxonomy of the extant dinoflagellate genus Gonyaulax is challenging since its thecate morphology is rather conservative. In contrast, cysts of Gonyaulax are varied in morphology and have been related with the fossil-based genera Spiniferites and Impagidinium. To better understand the systematics of Gonyaulax species, we performed germination experiments on cysts that can be identified as S. ristingensis, an unidentified Spiniferites with petaloid processes here described as Spiniferites pseudodelicatus sp. nov. and Impagidinium variaseptum from Chinese and Portuguese waters. Despite marked differences in cyst morphology, motile cells of S. pseudodelicatus and I. variaseptum are indistinguishable from Gonyaulax baltica. Motile cells hatched from S. ristingensis are morphologically similar to G. baltica as well but differ in the presence of one pronounced antapical spine. Three new species, Gonyaulax amoyensis (cyst equivalent S. pseudodelicatus), Gonyaulax bohaiensis (cyst equivalent I. variaseptum), and Gonyaulax portimonensis (cyst equivalent S. ristingensis), were erected. In addition, a new ribotype (B) of G. baltica was reported from South Korea and a bloom of G. baltica ribotype B is reported from New Zealand. Molecular phylogeny based on LSU and SSU rRNA gene sequences revealed that Gonyaulax species with minute or short antapical spines formed a well-resolved clade, whereas species with two pronounced antapical spines or lack of antapical spines formed the sister clade. Six strains of four above species were examined for yessotoxin production by liquid chromatography coupled with tandem mass spectrometry, and very low concentrations of yessotoxin were detected for one G. bohaiensis strain.

Taxonomy and Molecular Phylogenetics of Ensiculiferaceae, fam. nov. (Peridiniales, Dinophyceae), with Consideration of their Life-history.

In the current circumscription, the Thoracosphaeraceae comprise all dinophytes exhibiting calcified coccoid cells produced during their life-history. Species hitherto assigned to Ensiculifera and Pentapharsodinium are mostly based on the monadoid stage of life-history, while the link to the coccoid stage (occasionally treated taxonomically distinct) is not always resolved. We investigated the different life-history stages and DNA sequence data of Ensiculifera mexicana and other species occurring in samples collected from all over the world. Based on concatenated ribosomal RNA gene sequences Ensiculiferaceae represented a distinct peridinalean branch, which showed a distant relationship to other calcareous dinophytes. Both molecular and morphological data (particularly of the coccoid stage) revealed the presence of three distinct clades within Ensiculiferaceae, which may include other dinophytes exhibiting a parasitic life-history stage. At a higher taxonomic level, Ensiculiferaceae showed relationships to parasites and endosymbionts (i.e., Blastodinium and Zooxanthella) as well as to dinophytes harbouring diatoms instead of chloroplasts. These unexpected phylogenetic relationships are corroborated by the presence of five cingular plates in all such taxa, which differs from the six cingular plates of most other Thoracosphaeraceae. We herein describe Ensiculiferaceae, emend the descriptions of Ensiculifera and Pentapharsodinium, erect Matsuokaea and provide several new combinations at the species level.

Cryptic speciation in Protoceratium reticulatum (Dinophyceae): Evidence from morphological, molecular and ecophysiological data.

The cosmopolitan, potentially toxic dinoflagellate Protoceratium reticulatum possesses a fossilizable cyst stage which is an important paleoenvironmental indicator. Slight differences in the internal transcribed spacer ribosomal DNA (ITS rDNA) sequences of P. reticulatum have been reported, and both the motile stage and cyst morphology of P. reticulatum display phenotypic plasticity, but how these morpho-molecular variations are related with ecophysiological preferences is unknown. Here, 55 single cysts or cells were isolated from localities in the Northern (Arctic to subtropics) and Southern Hemispheres (Chile and New Zealand), and in total 34 strains were established. Cysts and/or cells were examined with light microscopy and/or scanning electron microscopy. Large subunit ribosomal DNA (LSU rDNA) and/or ITS rDNA sequences were obtained for all strains/isolates. All strains/isolates of P. reticulatum shared identical LSU sequences except for one strain from the Mediterranean Sea that differs in one position, however ITS rDNA sequences displayed differences at eight positions. Molecular phylogeny was inferred using maximum likelihood and Bayesian inference based on ITS rDNA sequences. The results showed that P. reticulatum comprises at least three ribotypes (designated as A, B, and C). Ribotype A included strains from the Arctic and temperate areas, ribotype B included strains from temperate regions only, and ribotype C included strains from the subtropical and temperate areas. The average ratios of process length to cyst diameter of P. reticulatum ranged from 15% in ribotype A, 22% in ribotype B and 17% in ribotype C but cyst size could overlap. Theca morphology was indistinguishable among ribotypes. The ITS-2 secondary structures of ribotype A displayed one CBC (compensatory change on two sides of a helix pairing) compared to ribotypes B and C. Growth response of one strain from each ribotype to various temperatures was examined. The strains of ribotypes A, B and C exhibited optimum growth at 15 °C, 20 °C and 20-25 °C, respectively, thus corresponding to cold, moderate and warm ecotypes. The profiles of yessotoxins (YTXs) were examined for 25 strains using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The parent compound yessotoxin (YTX) was produced by strains of ribotypes A and B, but not by ribotype C strains, which only produced the structural variant homoyessotoxin (homoYTX). Our results support the notion that there is significant intra-specific variability in Protoceratium reticulatum and the biogeography of the different ribotypes is consistent with specific ecological preferences.

Pentaplacodinium saltonense gen. et sp. nov. (Dinophyceae) and its relationship to the cyst-defined genus Operculodinium and yessotoxin-producing Protoceratium reticulatum.

Strains of a dinoflagellate from the Salton Sea, previously identified as Protoceratium reticulatum and yessotoxin producing, have been reexamined morphologically and genetically and Pentaplacodinium saltonense n. gen. et sp. is erected to accommodate this species. Pentaplacodinium saltonense differs from Protoceratium reticulatum (Claparède et Lachmann 1859) Bütschli 1885 in the number of precingular plates (five vs. six), cingular displacement (two widths vs. one), and distinct cyst morphology. Incubation experiments (excystment and encystment) show that the resting cyst of Pentaplacodinium saltonense is morphologically most similar to the cyst-defined species Operculodinium israelianum (Rossignol, 1962) Wall (1967) and O. psilatum Wall (1967). Collections of comparative material from around the globe (including Protoceratium reticulatum and the genus Ceratocorys) and single cell PCR were used to clarify molecular phylogenies. Variable regions in the LSU (three new sequences), SSU (12 new sequences) and intergenic ITS 1-2 (14 new sequences) were obtained. These show that Pentaplacodinium saltonense and Protoceratium reticulatum form two distinct clades. Pentaplacodinium saltonense forms a monophyletic clade with several unidentified strains from Malaysia. LSU and SSU rDNA sequences of three species of Ceratocorys (C. armata, C. gourreti, C. horrida) from the Mediterranean and several other unidentified strains from Malaysia form a well-supported sister clade. The unique phylogenetic position of an unidentified strain from Hawaii is also documented and requires further examination. In addition, based on the V9 SSU topology (bootstrap values >80%), specimens from Elands Bay (South Africa), originally described as Gonyaulax grindleyi by Reinecke (1967), cluster with Protoceratium reticulatum. The known range of Pentaplacodinium saltonense is tropical to subtropical, and its cyst is recorded as a fossil in upper Cenozoic sediments. Protoceratium reticulatum and Pentaplacodinium saltonense seem to inhabit different niches: motile stages of these dinoflagellates have not been found in the same plankton sample.

First record of resting cysts of the benthic dinoflagellate Prorocentrum leve in a natural reservoir in Gujan-Mestras, Gironde, France.

The resting cysts of the benthic dinoflagellate Prorocentrum leve from a natural reservoir in Gujan-Mestras (Gironde, France) were described in this study. The incubated urn-shaped cysts gave rise to cells of P. leve. Morphological observations through light microscopy and scanning electron microscopy, particularly of the periflagellar platelets, combined with large subunit ribosomal DNA sequences obtained through single-cell analysis confirm their affinity to the species P. leve. The cysts were characterized by a specific shape and the presence of an anterior plug. This is the first conclusive evidence for fossilizable resting stages within the Prorocentrales, one of the major orders within the Dinophyceae. Palynological treatments show that the cysts and endospores withstand hydrochloric and hydrofluoric acids. Micro-Fourier transform infrared analysis on single specimens suggests that the composition of the endospore is cellulosic and the cyst wall a more robust, noncellulosic ß-glucan. The spectra overall are similar to other published spectra of resting cysts from autotrophic, planktonic dinoflagellates.

Cryptic diversity within the harmful dinoflagellate Akashiwo sanguinea in coastal Chinese waters is related to differentiated ecological niches.

Blooms of the harmful dinoflagellate Akashiwo sanguinea are responsible for the mass mortality of fish and invertebrates in coastal waters. This cosmopolitan species includes several genetically differentiated clades. Four clonal cultures were established by isolating single cells from Xiamen Harbour (the East China Sea) for morphological and genetic analyses. The cultures displayed identical morphology but were genetically different, thus revealing presence of cryptic diversity in the study area. New details of the apical structure complex of Akashiwo sanguinea were also found. To investigate whether the observed cryptic diversity was related to environmental differentiation, 634 cells were obtained from seasonal water samples collected from 2008 to 2012. These cells were sequenced by single-cell PCR. For comparison with Chinese material, additional large subunit ribosomal DNA sequences were obtained for three established strains from Malaysian and French waters. To examine potential ecological differentiation of the distinct genotypes, growth responses of the studied strains were tested under laboratory conditions at temperatures of 12°C to 33°C. These experiments showed four distinct ribotypes of A. sanguinea globally, with the ribotypes A and B co-occuring in Xiamen Harbour. Ribotype A of A. sanguinea was present year-round in Xiamen Harbour, but it only bloomed in the winter and spring, thus corresponding to the winter type. In contrast, A. sanguinea ribotype B bloomed only in the summer, corresponding to the summer type. This differentiation supports the temperature optimum conditions that were established for these two ribotypes in the laboratory. Ribotype A grew better at lower temperatures compared to ribotype B which preferred higher temperatures. These findings support the idea that various ribotypes of A. sanguinea correspond to distinct ecotypes and allopatric speciation occurred in different climatic regions followed by dispersal.

Effect of nutrient pollution on dinoflagellate cyst assemblages across estuaries of the NW Atlantic.

We analyzed surface sediments from 23 northeast USA estuaries, from Maine to Delaware, and nine estuaries from Prince Edward Island (PEI, Canada), to determine how dinoflagellate cyst assemblages varied with nutrient loading. Overall the abundance of cysts of heterotrophic dinoflagellates correlates with modeled nitrogen loading, but there were also regional signals. On PEI cysts of Gymnodinium microreticulatum characterized estuaries with high nitrogen loading while the sediments of eutrophic Boston Harbor were characterized by high abundances of Spiniferites spp. In Delaware Bay and the Delaware Inland Bays Polysphaeridium zoharyi correlated with higher temperatures and nutrient loading. This is the first study to document the dinoflagellate cyst eutrophication signal at such a large geographic scale in estuaries, thus confirming their value as indicators of water quality change and anthropogenic impact.

Cyst-Theca Relationship and Phylogenetic Position of Impagidinium caspienense Incubated from Caspian Sea Surface Sediments: Relation to Gonyaulax baltica and Evidence for Heterospory within Gonyaulacoid Dinoflagellates.

We investigate the cyst-theca relationship of Impagidinium caspienense. Through an incubation experiment, we succeeded in examining the motile stage. Additional molecular analysis of single-cyst PCR (LSU and SSU rDNA) reveal that the cyst is related to the species Gonyaulax baltica Ellegaard et al. (). The ability of this species to belong to two types of cyst-based genera (spiniferate and impagidinioid) suggests that environmental (particularly salinity) and not genetic factors explain the formation of both morphotypes by G. baltica, which provides evidence for heterospory in this species. The affiliation to G. baltica demonstrates that I. caspienense is not endemic to the Caspian Sea. The phylogenetic position of several other gonyaulacoid species is also documented: Impagidinium pallidum, Ataxiodinium choane, Pyxidinopsis psilata, Spiniferites belerius, and Spiniferites ramosus. The LSU and SSU rDNA based phylogenies suggest that the genera Impagidinium and Spiniferites are not monophyletic, and that P. psilata and A. choane are close to Gonyaulax verior and Gonyaulax polygramma, respectively. In addition, this study accentuates the importance of cyst morphology in the classification of the Gonyaulacales.

Biogeography of dinoflagellate cysts in northwest Atlantic estuaries.

Few biogeographic studies of dinoflagellate cysts include the near-shore estuarine environment. We determine the effect of estuary type, biogeography, and water quality on the spatial distribution of organic-walled dinoflagellate cysts from the Northeast USA (Maine to Delaware) and Canada (Prince Edward Island). A total of 69 surface sediment samples were collected from 27 estuaries, from sites with surface salinities >20. Dinoflagellate cysts were examined microscopically and compared to environmental parameters using multivariate ordination techniques. The spatial distribution of cyst taxa reflects biogeographic provinces established by other marine organisms, with Cape Cod separating the northern Acadian Province from the southern Virginian Province. Species such as Lingulodinium machaerophorum and Polysphaeridinium zoharyi were found almost exclusively in the Virginian Province, while others such as Dubridinium spp. and Islandinium? cezare were more abundant in the Acadian Province. Tidal range, sea surface temperature (SST), and sea surface salinity (SSS) are statistically significant parameters influencing cyst assemblages. Samples from the same type of estuary cluster together in canonical correspondence analysis when the estuaries are within the same biogeographic province. The large geographic extent of this study, encompassing four main estuary types (riverine, lagoon, coastal embayment, and fjord), allowed us to determine that the type of estuary has an important influence on cyst assemblages. Due to greater seasonal variations in SSTs and SSSs in estuaries compared to the open ocean, cyst assemblages show distinct latitudinal trends. The estuarine context is important for understanding present-day species distribution, the factors controlling them, and to better predict how they may change in the future.

A new heterotrophic dinoflagellate from the North-eastern Pacific, Protoperidinium fukuyoi: cyst-theca relationship, phylogeny, distribution and ecology.

The cyst-theca relationship of Protoperidinium fukuyoi n. sp. (Dinoflagellata, Protoperidiniaceae) is established by incubating resting cysts from estuarine sediments off southern Vancouver Island, British Columbia, Canada, and San Pedro Harbor, California, USA. The cysts have a brown-coloured wall, and are characterized by a saphopylic archeopyle comprising three apical plates, the apical pore plate and canal plate; and acuminate processes typically arranged in linear clusters. We elucidate the phylogenetic relationship of P. fukuyoi through large and small subunit (LSU and SSU) rDNA sequences, and also report the SSU of the cyst-defined species Islandinium minutum (Harland & Reid) Head et al. 2001. Molecular phylogenetic analysis by SSU rDNA shows that both species are closely related to Protoperidinium americanum (Gran & Braarud 1935) Balech 1974. Large subunit rDNA phylogeny also supports a close relationship between P. fukuyoi and P. americanum. Three subgroups in total are further characterized within the Monovela group. The cyst of P. fukuyoi shows a wide geographical range along the coastal tropical to temperate areas of the North-east Pacific, its distribution reflecting optimal summer sea-surface temperatures of ~14-18 °C and salinities of 22-34 psu.

Environmental stress and recovery: the geochemical record of human disturbance in New Bedford Harbor and Apponagansett Bay, Massachusetts (USA).

Sediments record the history of contamination to estuaries. Analysis of the concentrations of toxic organic compounds, contaminant and crustal metals, organic carbon content and isotopic composition in sediment cores from two estuarine systems in Buzzards Bay allowed reconstruction of human impacts over 350 years. Vertical distributions of the contaminants correlate with changes in the nature of watershed/estuarine activities. All contaminants were highly enriched (tens to hundreds times background) in modern New Bedford Harbor sediments. Enrichment began around the turn of the 20th century for all but PCBs, which were first synthesized in the 1930s. An increase in organic carbon content and a shift of carbon isotopes toward a more terrestrial signature illustrates increasing anthropogenic impact in New Bedford as population grew along with the industrial base. Institution of environmental protection measures in the late 20th century was reflected in decreased, although still substantially elevated, concentrations of contaminants. A lack of industrial development in Apponagansett Bay resulted in much lower concentrations of the same indicators, although specific contaminants related to the early whaling industry increased significantly above background as early as the late 18th century. The similarity of indicators in older portions of cores from NBH and unimpacted Apponagansett Bay demonstrates that cores can be used to establish reference conditions as successfully as using separate sites judged a priori to represent the reference state. The historical reconstruction approach provides the basis for establishing relationships between environmental stressors and factors that drive the stressors, as well as a framework for the assessment of ecological response(s) to environmental stressors over a range of time and/or exposure scales.

Dinoflagellate cyst records and human disturbance in two neighboring estuaries, New Bedford Harbor and Apponagansett Bay, Massachusetts (USA).

The dinoflagellate cyst records in sediments from New Bedford Harbor and Apponagansett Bay demonstrate sensitivity to environmental change caused by human activity in the watersheds over the last 500 years. Changes in the species richness, as well as absolute and relative abundance of dinoflagellate cyst taxa reflect recent periods of development around the estuaries. Cyst taxa sensitive to these changes include Dubridinium spp., Polykrikos schwartzii, Lingulodinium machaerophorum, Operculodinium israelianum and Selenopemphix quanta. The greatest changes in the dinoflagellate cyst record occur during the 20th century, when New Bedford Harbor was exposed to both toxic pollution and heavy nutrient loading from point and non-point sources. Apponagansett Bay was not subject to industrial pollution and nutrient enrichment has been lower (from non-point sources). In Apponagansett Bay there is an increase in the dinoflagellate cyst species richness while species richness first increased, then declined in New Bedford Harbor. During the same period, the total dinoflagellate cyst concentration in New Bedford Harbor fluctuated over a wide range. The decline of species richness and the large fluctuations in the total cyst abundances signal the intensified anthropogenic disturbance in the watershed, notably a high degree of eutrophication and toxic pollution.