Microphytobenthic biomass assessment by pigment analysis: comparison of spectrophotometry and High Performance Liquid Chromatography methods

Samples of microphytobenthos from the Tagus estuary were analysed for photosynthetic pigments by spectrophotometry and High Performance Liquid Chromatography (HPLC). Chlorophyll a values obtained with HPLC and spectrophotometry methods presented a highly significant positive correlation for both spectrophotometric methods used (with and without the correction for pheopigments), but this relationship depended on the type of sediment. We concluded that spectrophotometric methods give reliable Chl-a values, being suited for routine analysis, when a vast number of replicates is needed. However, for the correct estimation of pheopigments, HPLC analysis is indispensable. In the literature, Chl-a estimations are expressed per content (μg g⁻¹) or concentration (mg m⁻²). We discuss the influence of sediment type on the results depending on the type of unit used, and propose a simple conversion factor based on sediment water content.     

Spatio-temporal distribution of the microphytobenthic biomass in intertidal flats of Tagus Estuary (Portugal)

A study on spatio-temporal distribution of microphytobhethos in intertidal zones of Tagus Estuary was carried out from 1990 to 1992. Near Lisbon, Portugal, Tagus Estuary is a shallow mesotidal estuary, covering an area of 320 km². The intertidal area ranges from 20 to 40% off the total area and it is constituted mainly by mudflats. Intertidal flats are richly populated by microalgae, diatoms being the most important and ubiquitos group.Spatial variation of microphytobethos was studied in spring 1990, 21 different sites were sampled. Microphytobenthos biomass was evaluated as chlorophyll a content of the surface centimeter, ranging from 10 to 240 mg m^–2. A Principal Component Analysis showed that 62% of the total variability found in intertidal flats of Tagus estuary could be attributed to two major factors: sediment type and tidal height. A hierarchical grouping defined 3 major groups of similar stations, each one representing a different strata of the ecosystem.One station from each group was chosen for the study of the temporal variation. A sampling, rogram took place from April 1991 to April 1992, with fortnightly sampling, the Chl a ranged from 20–300 mg m^–2. No clear seasonal variation was found, and our results indicated that tidal height of sampledsite played an essential role in temporal biomass evolution, thus upper littoral sites were influenced by climatic parameters, whereas in lower sites action of tides mainly controlled microphytic biomass.

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Résumé Une étude sur l'hétérogénéité spatio-temporelle du microphytobenthos dans les sédiments intertidaux de l'Estuaire du Tage a été accompli de 1990 á 1992.L'Estuaire du Tage, prés de Lisbonne (Portugal) est un estuaire peu profond, mesotidal, avec une aire total de 320 km². L'aire intertidale est comprise entre 20 et 40% du total, et constituéé surtout par des vasiéres. Ces slikkes sont peuplées par une communauté assez riche de microalgues, ou les diatomées sont les plus abundantes.La variation spatialle du microphytobenthos était évalué au Printemps 1990, ou 21 différentes stations étaient échantillonnées. La biomasse était évalué par la concentration enchlorophylle a du premier centimétre de sédiment, qui a varié de 10 á 240 mg Chl a m^–2. Une Analyse en Composants Principales a montré que 62% de la variabilité de la biomasse était lié á deux facteurs: le sédiment et l'hauteur vis-á-vis la marée. Une classification hiérarchique des stations par similitude a établi 3 groupes principaux, représantantles différents strates de écecosytéme.Une station de chaque groupement a été choisie pour l'étude de la variation temporelle, qui s'est deroulé d'avril 1991 á avril 1992, avec des prélévements deux fois par mois. Les valeurs de Chl a obtenus vont de 20 á 300 mg m^–2. Les variations saisonniéres observées ne sont pas claires: nos résultats indiquent que l'hauteur de la station (m) joue un rôle essentiel dans l'évolution temporel de la biomasse, c'est á dire, la biomasse microalgal des sites du supra-littoral est influencié par les paramétres climatiques, tandis que dans l'infra-littoral c'est l'action des marées le facteur principal.

     

Short-term effects of winter warming and acidification on phytoplankton growth and mortality: more losers than winners in a temperate coastal lagoon

Hydrobiologia - 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...     

Monitoring Migration and Measuring Biomass in Benthic Biofilms: The Effects of Dark/far-red Adaptation and Vertical Migration on Fluorescence Measurements

Pulse modulated fluorescence has increasingly been used as an ecological tool to examine changes in the vertical distribution of microphytobenthic cells within the upper layers of estuarine sediments (most often using the minimum fluorescence yield F(o)) as well as to indicate the health of the community (using the maximum PS II quantum efficiency F(v)/F(m)). However, the practicalities of in situ measurements, often dictates that short dark adaptation periods must be used ( approximately 15 min). The use of far-red light as an alternative to dark adaptation was investigated in natural migratory microphytobenthic biofilms and artificial non-migratory biofilms. Prolonged periods of darkness ( approximately 24 h) were not adequate to achieve 'true' measurements of F(o) and F(v)/F(m), which require complete oxidation of Q(A) and full reversal of non-photochemical quenching (NPQ). In some instances, stable values were only achieved using far-red light. Prolonged exposure to dark/far-red light led to a downwards migration of cells in natural assemblages, as seen by a reduction in both F(o) and the maximum fluorescence yield (F(m)). In non-migratory biofilms, F(m) increased in the dark and far-red treatments, indicating a reversal of NPQ, whereas F(o) decreased in far-red light but increased in the dark. It is suggested that far-red light and darkness differentially affected the balance between NPQ reversal and Q(A) oxidation that lead to the measured F(o) yield. The use of far-red light as an alternative to dark adaptation is discussed and the implications of short (e.g., 15 min) dark adaptation times used in situ are discussed with reference to the vertical migration of cells within sediment biofilms.     

Variability in Chlorophyll and Phytoplankton Composition in an Estuarine System

The spatial and temporal variability of phytoplankton abundance (in terms of chlorophyll a and cell number), inorganic nitrogen, suspended particulate matter (SPM), and light availability was determined throughout one year in the Tagus estuary, Portugal. Chlorophyll a concentrations showed a strong seasonal variation with values ranging from 1 to 32 μg l^?1(average 5.4 μg l^?1). Chlorophyll patterns were unimodal for sites 1, 2, and 3 and bimodal for site 4. Diatoms and cryptophytes were, throughout the year, the dominant groups in this shallow and unstratified estuarine system. Nitrate concentrations were seasonally related to river flow and ammonium concentrations spatially related to sources of sewage input. Lower river inputs and long water residence times during summer initially promoted the accumulation of phytoplankton, but the resulting low dissolved inorganic nitrogen (DIN) concentrations lead to limitation of phytoplankton growth. Chlorophyll a and DIN values obtained in the present study were comparable to those reported 20 years ago for the Tagus estuary.     

Influence of drought on algal biofilms and meiofaunal assemblages of temperate reservoirs and rivers

The role of hydrological droughts in shaping meiofauna abundance through alterations in biofilm biomass and composition was investigated. In January 2005, continental Portugal was under a moderate to severe drought resulting from a 40% to 60% decrease in rainfall during the previous 12 months relative to the long-term average (1961–1990). Reservoir capacity was reduced by 30–50% relative to average values and the width of streams was reduced by 20–80% in the Zêzere River Basin (central Portugal). Algal biomass and algal class composition of biofilms was assessed through quantification of algal pigments in three reservoir and six river locations. During drought, habitat alterations are expected to be sharp in rivers while, in the absence of water quality deterioration, the habitat characteristics of reservoirs are expected to remain fairly unaffected. Chlorophylls and carotenoid pigments were extracted from biofilm samples and analysed using high performance liquid chromatography (HPLC). In the winter of 2003, during the period of average rainfall, biofilm biomass did not exceed 5 μg chlorophyll a cm⁻² at any location. River biofilm biomass was roughly half of that measured in the reservoirs. In the winter of 2005 (drought), biofilm biomass increased by more than 5-fold in river locations and remained low or decreased in the reservoirs. Algal biofilms were either dominated by Bacillariophyceae or by Chlorophyceae regardless of the existence of drought. The relative contribution of Bacillariophyceae to total biofilm biomass was higher during the drought than under average hydrological conditions. The abundance of harpacticoids, cladocerans and ostracods was favoured by the drought only in the reservoirs where an increase in diatom proportion in biofilms was observed. The increase in the abundance of cyclopoid copepods, turbellarians, nematodes and chironomids in rivers during the drought could be explained by algal class composition and biomass of biofilms and environmental variables (organic matter sediment content, phosphorus availability content and sediment granulometry). The hydrological drought appears to regulate meiofauna abundance only in river locations, possibly through the promotion of the growth of biofilms and the availability of organic matter deposited in rivers during the drought. Handling editor: D. Ryder     

Oxygen profiles in intertidal sediments of Ria Formosa (S. Portugal)

Microelectrode oxygen profiles were measured in intertidal sediments from Ria Formosa (S. Portugal), a very productive shallow coastal lagoon. Four intertidal sampling sites were selected according to different sediment characteristics. Individual profiles revealed a high degree of lateral variability on a centimeter spatial scale. Nevertheless, consistent differences were observed between oxygen profiles measured in atmosphere-exposed and inundated intertidal sediments: in organically poor sand oxygen-penetration depth varied from 3 mm in inundated cores to more than 7 mm in exposed ones, while in organically rich muddy sand and mud it remained between 0.5–2.0 mm. The oxygen input from inundated to exposed conditions was estimated for each sampling site. Semi-diurnal tidal fluctuation, leading to periodical atmospheric exposure of sediments plays a major role in the oxygenation process of intertidal zones of Ria Formosa.     

Structure and diversity of intertidal benthic diatom assemblages in contrasting shores: a case study from the Tagus estuary1

The structure of intertidal benthic diatoms assemblages in the Tagus estuary was investigated during a 2‐year survey, carried out in six stations with different sediment texture. Nonparametric multivariate analyses were used to characterize spatial and temporal patterns of the assemblages and to link them to the measured environmental variables. In addition, diversity and other features related to community physiognomy, such as size‐class or life‐form distributions, were used to describe the diatom assemblages. A total of 183 diatom taxa were identified during cell counts and their biovolume was determined. Differences between stations (analysis of similarity (ANOSIM), R = 0.932) were more evident than temporal patterns (R = 0.308) and mud content alone was the environmental variable most correlated to the biotic data (BEST, ρ = 0.863). Mudflat stations were typically colonized by low diversity diatom assemblages (H′ ~ 1.9), mainly composed of medium‐sized motile epipelic species (250–1,000 μm³), that showed species‐specific seasonal blooms (e.g., Navicula gregaria Donkin). Sandy stations had more complex and diverse diatom assemblages (H′ ~ 3.2). They were mostly composed by a large set of minute epipsammic species (<250 μm³) that, generally, did not show temporal patterns. The structure of intertidal diatom assemblages was largely defined by the interplay between epipelon and epipsammon, and its diversity was explained within the framework of the Intermediate Disturbance Hypothesis. However, the spatial distribution of epipelic and epipsammic life‐forms showed that the definition of both functional groups should not be over‐simplified.     

Diatom migration and sediment armouring – an example from the Tagus Estuary, Portugal

This study measured sediment stability, colloidal and total carbohydrate and chlorophyll a in the upper 2 mm of the sediment over a whole emersion period (0800–1140) in the Tagus estuary, Portugal on the 18th April 2000. Low-temperature scanning electron microscopy (LTSEM) time series images revealed the migration of microalgae throughout the emersion period, including their appearance at the sediment surface at the beginning of emersion and their subsequent return below the surface at the end of emersion. Different species arrived at the sediment surface at different times and there was a slight lag between the appearance of the first cells and the subsequent increase in sediment stability. Increased chlorophyll a concentration in the surface sediments preceded the increase in sediment stability, whilst colloidal and total carbohydrate concentrations increased afterwards. Sediment water content decreased during the emersion period. Erosion threshold increased shortly after the microalgal cells appeared at the sediment surface, suggesting that the cells themselves act to `armour' the sediment surface, retarding erosion. Lack of correlation between sediment stability and factors traditionally considered to control sediment stability (e.g., water content and carbohydrate content) indicates that an important variable or interaction has yet to be identified. One possibility is that the carbohydrate fraction extracted does not measure accurately the binding effectiveness of the carbohydrates in the sediment. We propose feedback and `critical point' models to explain how the various sediment properties determine sediment stability. The implication is that sediment stability varies in an apparently idiosyncratic and site-specific fashion due to the complex interaction of physical and biological variables. Given the importance of ecological processes in intertidal sediments, the measurement, understanding and modelling of sediment erosion would benefit greatly from the application of ecological methods of experimental design and sampling.     

A mathematical model for the vertical distribution of chlorophyllA in estuarine intertidal sediments

Coastal and estuarine intertidal sediments are commonly colonized by dense populations of microphytobenthos. Due to wind and tides, important fractions of microphytobenthic populations may be buried. A mathematical model describing the depth variation of chlorophyll a in intertidal sediments was developed and experimentally tested. The model assumed first-order chlorophylla degradation and a constant mean burial velocity which resulted in a negative exponential variationC _Z =C _O exp{-k/vz} (C _Z andC _O =chlorophylla concentration at depth zand at the surface;k=specific degradation rate of chlorophyll a to pheopigments;V=mean burial velocity). Chlorophylla concentration depth profiles in different sediment types measured at the Tagus estuary and Ria Formosa (Portugal) were used to validate the model. The model was adjusted to field data. The chlorophyll a degradation rate was measured in a microcosm experiment under total darkness and no tidal action, and sampled during three months. This rate was shown to be independent of time and depth for the upper 0–15 mm depth interval. This result allowed the estimation ofV for each sampling site. Comparison of predicted and observed temporal data further confirmed the validity of the model andk andV values. Despite its simplicity, the proposed model adequately described the depth distribution of chlorophylla in different types of intertidal sediments. The model allowed the quantitative characterization of the buried microphytobenthic biomass (depth-integrated biomass) and the assessment of its importance as potentially productive stock of cells.