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Hematopoietic stem cells (HSCs) produce all essential cellular components of the blood. Stromal cell lines supporting HSCs follow a vascular smooth muscle cell (vSMC) differentiation pathway, suggesting that some hematopoiesis-supporting cells originate from vSMC precursors. These pericyte-like precursors were recently identified in the aorta-gonad-mesonephros (AGM) region; however, their role in the hematopoietic development in vivo remains unknown. Here, we identify a subpopulation of NG2+Runx1+ perivascular cells that display a sclerotome-derived vSMC transcriptomic profile. We show that deleting Runx1 in NG2+ cells impairs the hematopoietic development in vivo and causes transcriptional changes in pericytes/vSMCs, endothelial cells and hematopoietic cells in the murine AGM. Importantly, this deletion leads also to a significant reduction of HSC reconstitution potential in the bone marrow in vivo. This defect is developmental, as NG2+Runx1+ cells were not detected in the adult bone marrow, demonstrating the existence of a specialised pericyte population in the HSC-generating niche, unique to the embryo.Hematopoietic stem cells are supported by niche cells that help balance stem cell self-renewal and differentiation. Here they show that Runx1 deletion in the embryonic perivascular HSC niche impairs hematopoietic development in vivo and causes transcriptional changes in pericytes/vSMCs, endothelial cells and hematopoietic cells in the murine AGM. [ABSTRACT FROM AUTHOR]

Hematopoietic stem cells (HSCs) produce all essential cellular components of the blood. Stromal cell lines supporting HSCs follow a vascular smooth muscle cell (vSMC) differentiation pathway, suggesting that some hematopoiesis-supporting cells originate from vSMC precursors. These pericyte-like precursors were recently identified in the aorta-gonad-mesonephros (AGM) region; however, their role in the hematopoietic development in vivo remains unknown. Here, we identify a subpopulation of NG2+Runx1+ perivascular cells that display a sclerotome-derived vSMC transcriptomic profile. We show that deleting Runx1 in NG2+ cells impairs the hematopoietic development in vivo and causes transcriptional changes in pericytes/vSMCs, endothelial cells and hematopoietic cells in the murine AGM. Importantly, this deletion leads also to a significant reduction of HSC reconstitution potential in the bone marrow in vivo. This defect is developmental, as NG2+Runx1+ cells were not detected in the adult bone marrow, demonstrating the existence of a specialised pericyte population in the HSC-generating niche, unique to the embryo. Hematopoietic stem cells are supported by niche cells that help balance stem cell self-renewal and differentiation. Here they show that Runx1 deletion in the embryonic perivascular HSC niche impairs hematopoietic development in vivo and causes transcriptional changes in pericytes/vSMCs, endothelial cells and hematopoietic cells in the murine AGM. [ABSTRACT FROM AUTHOR]

Phytoplankton are dominant primary producers and key indicators in aquatic ecosystems. Understanding the controlling factors on the structure of phytoplankton assemblages is fundamental, but particularly challenging at the land–ocean interface. To identify the patterns and predictors of phytoplankton assemblage structure in the Ria Formosa coastal lagoon (south Portugal), this study combined phytoplankton abundance along a transect between the discharge point of a wastewater treatment plant and a lagoon inlet, over two years, with physico-chemical, hydrographic, and meteo-oceanographic variables. Our study identified 147 operational taxonomic units (OTUs), and planktonic diatoms (60–74%) and cryptophyceans (17–25%) dominated the phytoplankton in terms of abundance. Despite strong lagoon hydrodynamics, and the lack of spatial differences in the phytoplankton abundance and most diversity metrics, the multivariate analysis revealed differences in the assemblage structure between stations (p < 0.001) and seasons (p < 0.01). Indicator analysis identified cryptophyceans as lagoon generalists, and 11 station-specific specialist OTUs, including Kryptoperidinium foliaceum and Oscillatoriales (innermost stations) and potentially toxigenic species (Pseudo-nitzschia and Dinophysis; outer lagoon station, p < 0.05). Water temperature, pH, and nutrients emerged as the variables that best explained the changes in the phytoplankton assemblage structure (p < 0.001). Our findings provide insight into the relevance of local anthropogenic and natural forcings on the phytoplankton assemblage structure and can be used to support the management of RF and other coastal lagoons. [ABSTRACT FROM AUTHOR]

Phytoplankton play a key role in marine ecosystems, making the accurate quantification of functional groups/species, using standardized microscopy techniques, essential in many research efforts. In this study we aimed to evaluate the effects of (a) storage time on the abundance of microphytoplankton in samples fixed with acid Lugol's solution; (b) storage time on the abundance of pico- and nanophytoplankton in frozen microscopy slides; (c) storage time on the abundance of pico- and nanophytoplankton in fixed refrigerated samples and (d) storage protocol (refrigerated versus frozen) on the abundance of pico- and nanophytoplankton. Microphytoplankton were analyzed using inverted microscopy, and pico- and nanophytoplankton using epifluorescence microscopy. Results indicate storage time negatively impacted the abundances of all phytoplankton size classes; however, effects were group-specific. For accurate abundance estimates, we suggest that samples fixed with acid Lugol's should be analyzed within 30 and 180 days, for dinoflagellates/total microphytoplankton and diatoms, respectively. For picoeukaryotes, glutaraldehyde fixed samples should be kept refrigerated for up to 4 days, and slides should be prepared immediately before observation. It is recommended that authors specify the exact lag times between sample collection, fixation and analysis, to allow the comparability of phytoplankton datasets across different studies and/or monitoring programs. [ABSTRACT FROM AUTHOR]

Planktonic heterotrophic prokaryotes (HProks) are a pivotal functional group in marine ecosystems and are highly sensitive to environmental variability and climate change. This study aimed to investigate the short-term effects of increasing carbon dioxide (CO2), ultraviolet radiation (UVR), and temperature on natural assemblages of HProks in the Ria Formosa coastal lagoon during winter. Two multi-stressor microcosm experiments were used to evaluate the isolated and combined effects of these environmental changes on HProk abundance, production, growth, and mortality rates. The isolated and combined effects of increased CO2 on HProks were not significant. However, HProk production, cellular activity, instantaneous growth rate, and mortality rate were negatively influenced by elevated UVR and positively influenced by warming. Stronger effects were detected on HProk mortality in relation to specific growth rate, leading to higher HProk net growth rates and abundance under elevated UVR and lower values under warming conditions. [ABSTRACT FROM AUTHOR]

The Ria Formosa coastal lagoon is a highly productive shallow ecosystem in southern Portugal, subjected to nutrient inputs from anthropogenic and natural sources. Nutrients are major abiotic drivers of phytoplankton in this system, but their effects on phytoplankton assemblages and the occurrence of nutrient limitation are still poorly understood. The main goal of this study was, thus, to evaluate the occurrence, type, and effects of nutrient limitation on phytoplankton community and specific functional groups in the Ria Formosa coastal lagoon. We conducted nutrient enrichment experiments with factorial additions of nitrogen (N) and phosphorus (P) using natural phytoplankton assemblages from distinct locations in the Ria Formosa, throughout a yearly cycle. Phytoplankton composition and abundance were evaluated using inverted and epifluorescence microscopies, and spectrophotometric methods were used for biomass. Limitation was defined as higher phytoplankton growth following enrichment with a particular nutrient in relation to the non-enriched control. The most common type of phytoplankton limitation was simultaneous co-limitation by N and P; diatoms, as r-strategists, were the most frequently limited group. Single N and P limitation, and serial P limitation were also observed, as well as negative responses to nutrient enrichment. Group-specific responses to nutrient enrichment were not reflected in the relative abundance of phytoplankton groups within the whole assemblage, due to the numerical dominance of pico-sized groups (cyanobacteria and eukaryotic picophytoplankton). Ambient nutrient ratios and concentrations did not predict phytoplankton nutrient limitation, given the different nutrient utilisation traits among phytoplankton functional groups. Therefore, nutrient ratios should not be used as indicators of nutrient limitation in eutrophication assessment. [ABSTRACT FROM AUTHOR]

The use of several light-related variables, such as the Secchi disc depth, the euphotic depth, and in particular, the diffuse attenuation coefficient (Kd), is deeply rooted in phytoplankton research, but these are not the most appropriate indicators of the amount of light available for photosynthesis. We argue that the variable of interest for phytoplankton is the mean light intensity in the mixed layer (Im), which represents the mean light to which phytoplankton cells are exposed throughout their life cycle, while being continuously mixed in the mixed layer. We use empirical data collected in different coastal ecosystems in southern Portugal to demonstrate why Im should be the preferred metric instead of the deeply rooted Kd. We show that, although the relationship between Im and Kd is inversely proportional, it is not always strong or even significant. Different Im values can be associated with the same Kd, but distinct Im have different physiological effects of phytoplankton. Therefore, Kd does not capture the amount of light available for photosynthesis, given that, unlike Im, Kd calculation does not consider the depth of the mixed layer. Therefore, we urge phytoplankton researchers to consider the measurement and calculation of Im when evaluating light-related processes in phytoplankton ecology. [ABSTRACT FROM AUTHOR]

It is generally assumed that the larger the bottle volume, the longer the duration of phytoplankton microcosm experiments. We hypothesize that volume and duration are independent, as volume does not regulate the extension of the exponential growth phase. We conducted two microcosm experiments using 1, 2, and 8 L bottles, inoculated with phytoplankton collected in the Ria Formosa lagoon (SE Portugal) and incubated for 1, 2, 4, and 8 days. Phytoplankton net growth rates were estimated using chlorophyll a concentration and cell abundance, determined with epifluorescence and inverted microscopy. Results show that the experimental duration significantly affected net growth rates, independently of volume, with decreasing net growth rates with time. Regarding volume, we found significant, but weak, differences in net growth rates, and significant two-way interactions only for the larger-sized cells. No significant differences in net growth rates across the different volumes were detected for the smaller, most abundant taxa and for the whole assemblage. We conclude that duration, not volume, is the main factor to consider in microcosm experiments, and it should allow the measurement of responses during the exponential growth phase, which can be detected through daily sampling throughout the duration of the experiment. [ABSTRACT FROM AUTHOR]

Changes in temperature and CO2 are typically associated with climate change, but they also act on shorter time scales, leading to alterations in phytoplankton physiology and community structure. Interactions among stressors may cause synergistic or antagonistic effects on phytoplankton dynamics. Therefore, the main goal of this work is to understand the short-term isolated and interactive effects of warming and high CO2 on phytoplankton nutrient consumption, growth, production, and community structure in the Ria Formosa coastal lagoon (southern Portugal). We performed microcosm experiments with temperature and CO2 manipulation, and dilution experiments under temperature increase, using winter phytoplankton assemblages. Phytoplankton responses were evaluated using inverted and epifluorescence microscopy. Overall, phytoplankton growth and microzooplankton grazing on phytoplankton decreased with warming. Negative antagonist interactions with CO2 alleviated the negative effect of temperature on phytoplankton and cryptophytes. In contrast, higher temperature benefited smaller-sized phytoplankton, namely cyanobacteria and eukaryotic picophytoplankton. Diatom growth was not affected by temperature, probably due to nutrient limitation, but high CO2 had a positive effect on diatoms, alleviating the effect of nutrient limitation. Results suggest that this winter phytoplankton assemblage is well acclimated to ambient conditions, and short-term increases in temperature are detrimental, but can be alleviated by high CO2. [ABSTRACT FROM AUTHOR]

Jacob, J.; Correia, C.; Torres, A.F.; Xufre, G.; Matos, A.; Ferreira, C.; Reis, M.P.; Caetano, S.; Freitas, C.S.; Barbosa, A.B., and Cravo, A., 2020. Impacts of decommissioning and upgrading urban wastewater treatment plants on the water quality in a shellfish farming coastal lagoon (Ria Formosa, South Portugal). In: Malvárez, G. and Navas, F. (eds.), Global Coastal Issues of 2020. Journal of Coastal Research, Special Issue No. 95, pp. 45–50. Coconut Creek (Florida), ISSN 0749-0208. Ria Formosa is a productive coastal lagoon, located on the south coast of Portugal, and represents the largest national producer of shellfish bivalves (ca. 90% production). This ecosystem is subjected to various anthropogenic pressures, including the discharge of urban wastewater treatment plants (UWWTP), which impacts the lagoon water quality. This study aimed to assess the impact of alterations in the functioning of two UWWTP on the water quality of Ria Formosa, based on chemical variables, phytoplankton composition (including potential harmful species) and faecal contamination. During the period September 2018 - October 2019, water sampling was conducted along dominant longitudinal gradients of the effluent dispersion from the discharge point (1-2 km), for two sites: a decommissioned (OP) and a modified (FO) UWWTP. After modification, the later started receiving a higher influent volume (ca. 40%), under an innovative technology system (biological treatment in aerobic granular sludge). Based on chemical water quality variables, phytoplankton and indicators of faecal contamination, a significant improvement along the longitudinal gradient from the discharge point was observed after OP decommissioning. This improvement was fast, being detected two months after decommissioning, positively affecting areas used as shellfish farming grounds. However, distribution patterns of bacteriological indicators and regular shellfish harvesting interdictions suggested an alternative source of faecal contamination after OP decommissioning. At FO, both chemical variables and bacteriological indicators of faecal contamination revealed a slower improvement, only six-months after the UWWTP alteration. Before that, increased and highly variable ammonium, chlorophyll a concentration, phytoplankton abundances and Escherichia coli densities, revealed an unstable phase. Overall, a lower water quality at FO in respect to OP reflected not only a higher effluent volume but also more restricted water circulation for the former. [ABSTRACT FROM AUTHOR]

Jellyfish modulate the energy fluxes between compartments of aquatic ecosystems and the period that energy remains within each compartment. However, the contribution of different sources of organic matter (OM) to the production of most estuarine jellyfish species is unknown. Therefore, this study aimed to identify and quantify the putative sources of OM for Blackfordia virginica (Cnidaria, Hydrozoa) Mayer, 1910 in two temperate estuaries, based on the analyses of carbon and nitrogen stable isotope ratios. Zooplankton was the main source of OM assimilated by B. virginica in both Mira and Guadiana estuaries. However, particulate organic matter (POM) also contributed to B. virginica biomass, up to 26 and 37% in Mira and Guadiana estuaries, respectively. The POM pool was apparently comprised by terrestrialderived OM, as based on stable isotope ratios and C:NPOM. Terrestrial-derived OM is an undescribed carbon source for estuarine jellyfish. Our results support a working hypothesis stating that terrestrial-derived OM might support good physiological condition of B. virginica during periods of low metazooplankton abundance, through a detritusbased microbial food web transformation. Subsequent studies on trophic pathways ought to evaluate the temporal contribution of autochthonous and allochthonous OM and its consequences, accounting for the connectivity across ecosystems (terrestrial-estuarine-marine) and the interactions within compartments (pelagic-benthic). [ABSTRACT FROM AUTHOR]

The feeding ecology of Blackfordia virginica was evaluated concurrently with their ecophysiological condition in a temperate estuary. The diet of B. virginica is composed not only of metazooplankton, as commonly observed for other jellyfish species, but also of phytoplankton, ciliates and detritus. This feeding behavior might explain their good nutritional condition and sustainable growth during bloom peaks, when zooplankton abundance has already decreased significantly. [ABSTRACT FROM AUTHOR]

Increases in ultraviolet radiation (UVR) and CO2 affect phytoplankton growth and mortality in a variety of different ways. However, in situ responses of natural phytoplankton communities to climate change, as well as its effects on phytoplankton annual cycles, are still largely unknown. Although temperature and UVR have been increasing in temperate latitudes during winter, this season is still particularly neglected in climate change studies, being considered a non-active season regarding phytoplankton growth and production. Additionally, coastal lagoons are highly productive ecosystems and very vulnerable to climate change. This study aims, therefore, to evaluate the short-term effects of increased UVR and CO2 on the composition and growth of winter phytoplankton assemblages in a temperate coastal lagoon. During winter 2012, microcosm experiments were used to evaluate the isolated and combined effects of UVR and CO2, under ambient and high CO2 treatments, exposed to ambient UV levels and photosynthetically active radiation (PAR), or to PAR only. Phytoplankton composition, abundance, biomass and photosynthetic parameters were evaluated during the experiments. Significant changes were observed in the growth of specific phytoplankton groups, leading to changes in community composition. The cyanobacterium Synechococcus was dominant at the beginning of the experiment, but it was negatively affected by UVR and CO2. Diatoms clearly benefited from high CO2 and UVR, particularly Thalassiosira. Despite the changes observed in specific phytoplankton groups, growth and production of the whole phytoplankton community did not show significant responses to UVR and/or CO2. [ABSTRACT FROM AUTHOR]

The effects of different phases of semidiurnal and spring-neap tidal cycles on phytoplankton and environmental drivers were evaluated in a tidal, freshwater location of a mesotidal estuary (Guadiana estuary, SW Iberia). An Eulerian approach was used and sampling covered different seasons during 2008. Samples were collected during spring and neap tides, at high tide, mid-ebb, low tide and mid-flood. Several physical-chemical variables were measured, as well as phytoplankton abundance and biomass. Salinity was higher at high-tide and suspended particulate matter was higher during spring tides and flood, due to higher vertical mixing and resuspension of bottom sediments. Chlorophyll a concentration during winter and summer neap tides was higher than during spring tides, whilst the abundance of pennate diatoms was higher during winter and Spring spring tides than during neap tides, probably reflecting differences in river discharge. Overall, tidally-induced differences detected in the freshwater tidal reaches of the Guadiana estuary were not as considerable as those observed in the lower estuary. However, the occurrence of tidally-induced variability in some seasons reflects that thorough sampling programs to study estuarine tidal dynamics should be conducted throughout the year. Occasional sampling will not reflect the typical variability of these highly dynamic systems. (© 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [ABSTRACT FROM AUTHOR]

Phytoplankton seasonal and interannual variability in the Guadiana upper estuary was analyzed during 1996-2005, a period that encompassed a climatic controlled reduction in river flow that was superimposed on the construction of a dam. Phytoplankton seasonal patterns revealed an alternation between a persistent light limitation and episodic nutrient limitation. Phytoplankton succession, with early spring diatom blooms and summer-early fall cyanobacterial blooms, was apparently driven by changes in nutrients, water temperature, and turbulence, clearly demonstrating the role of river flow and climate variability. Light intensity in the mixed layer was a prevalent driver of phytoplankton interannual variability, and the increased turbidity caused by the Alqueva dam construction was linked to pronounced decreases in chlorophyll a concentration, particularly at the start and end of the phytoplankton growing period. Decreases in annual maximum and average abundances of diatoms, green algae, and cyanobacteria were also detected. Furthermore, chlorophyll a decreases after dam filling and a decrease in turbidity may point to a shift from light limitation towards a more nutrient-limited mode in the near future. [ABSTRACT FROM AUTHOR]