Usefulness of Simultaneous Use of Several Methods for the Estimation of Phytoplanktonic Biomass

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The annual cycle of phytoplankton biomass was followed in a eutrophic lake (Lake Aydat, Massif Central, France), using classic descriptors (biovolumes and chlorophyll a) as well as adenosine-5'-phosphate (ATP) levels. ATP is the metabolite indicating living biomass. ATP/Cell count and ATP/Chlorophyll a ratios were elevated due to the presenceof heterotrophic organisms, such as several species of ciliates. The first ratio is greater than the second. The difference can be explained either by an underestimation of the nanoplankton fraction through the cell count method, or by a lack of sedimentation in the settling chambers used for counting cells.

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... Some of the estimated flows were measured during previous studies that focused on spring blooms in Lake Pavin and are introduced as additional equations (lines 12-15 of Table 1); these include values for total gross and net primary production (Devaux, 1980;Bettarel et al., 2003), bacterial production (Bettarel et al., 2003) and viral lysis of bacteria (Bettarel et al., 2003) considered as the value of the flux from bacteria to DOC. Some other equalities were introduced for the Aydat model and values of total gross primary production (Aleya et al., 1988), bacterivory by heterotrophic nanoflagellates and microzooplankton (Bettarel et al., 2004) were considered (lines 16-18 of Table 1). Primary production values used for Pavin and Aydat were measured from 14C uptake according to Steemann-Nielsen (1952). ...
... mgC m −2 d −1 , March to June 2007). These values corroborate previous findings on the trophic status of both an oligo-mesotrophic lake (Amblard et al., 1992) and a eutrophic lake (Aleya et al., 1988). These values are consistent with gross primary production measured during a study conducted during spring phytoplankton proliferation in the north basin of the mesotrophic Japanese Lake Biwa (1639 mgC m −2 d −1 ; Yoshimizu et al., 2001). ...
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This study assesses the quantitative impact of parasitic chytrids on the planktonic food web of two contrasting freshwater lakes during different algal bloom situations. Carbon-based food web models were used to investigate the effects of chytrids during the spring diatom bloom in Lake Pavin (oligo-mesotrophic) and the autumn cyanobacteria bloom in Lake Aydat (eutrophic). Linear inverse modeling was employed to estimate undetermined flows in both lakes. The Monte Carlo Markov chain linear inverse modeling procedure provided estimates of the ranges of model-derived fluxes. Model results confirm recent theories on the impact of parasites on food web function through grazers and recyclers. During blooms of "inedible" algae (unexploited by planktonic herbivores), the epidemic growth of chytrids channeled 19-20% of the primary production in both lakes through the production of grazer exploitable zoospores. The parasitic throughput represented 50% and 57% of the zooplankton diet, respectively, in the oligo-mesotrophic and in the eutrophic lakes. Parasites also affected ecological network properties such as longer carbon path lengths and loop strength, and contributed to increase the stability of the aquatic food web, notably in the oligo-mesotrophic Lake Pavin.
Biomass in size-fractionated plankton (0.2–1.2 μm; 1.2–12/μm; 12–45 μm and 45–160 μm), together with that of ciliated protozoans (size fraction 1–160 μm), were assessed during stratification, in an eutrophic lake (Lake Aydat, Massif Central, France). Samples (51) were gathered from depths of 2, 7 and 14 m during 17 weekly samplings in the centre of the lake. Three biomass estimations were performed: cell-counts, chlorophyll a and ATP assays. Ciliates were counted live. Five species groups dominated the ciliate community: Loxodes striatus, Frontonia spp., Spirostomum ambiguum, Strombidium spp. and Dexiotrica plagia. Frontonia always had the highest cell concentration (54–64% of total cell counts), while the bulk of biomass was divided between Loxodes (42–70% of total) and Frontonia (29–53% of total). In terms of carbon, both the bacteria and ciliate contributions to the total microplanktonic biomass (size fraction 1–160 μm, i.e. bacteria + phytoplankton + ciliates) depends on the choice of the phytoplankton biomass estimation technique used. Thus, the contributions increase respectively from 16% and 19%, when phytoplankton biomass is estimated from chlorophyll α concentrations, to 24% and 27%, when phytoplankton biomass is based on biovolume. Relationships between bacteria, phytoplankton and ciliated protozoans reveal that ATP data are greatly overestimated by bacteria and ciliate interferences, especially in the metalimnion and hypolimnion. In the epilimnion, this interference is less. Interpretations of phytoplankton dynamics and metabolic activity from ATP assays should be made with extreme care, especially in eutrophic environments.
Three methods of measuring phytoplankton biomass (microscopic counting, electronic particle counting and determination of chlorophyll a concentration) were compared on both a daily (4 days) and a seasonal basis, in Charnwood Water, a small English freshwater lake. Correlations among measures were generally poor within days. However, good correlations were achieved among all methods on a seasonal basis. Seasonal correlations, in particular those between total particle numbers from particle counting and algal numbers from microscope, were affected by the degree of stability of the water column, with different relationships being found for mixed periods compared to stratified periods. These differences were related to an increased amount of particulate matter affecting the total particle numbers estimate during mixed periods. Other workers have found better relationships among these phytoplankton biomass methods within short periods, but there appears to be considerable variability among lakes. Therefore, it is recommended that the most appropriate method be evaluated on a individual lake basis, depending on the aims of the study.
Short-term and spatial fluctuations in specific biovolumes (volume x cell−1) of different morphological categories of planktonic bacteria were estimated microscopically. Samples were taken from two lakes occurring in two different climatic systems: Lake Aydat (France) and Lake Cromwell (Canada). The study was done in summer, using 24-hour cycles of sampling. Due to their large size, the specific volume of filamentous bacteria constituted, on average, the major part (>70%) of the total specific volume of all bacterial forms considered. Greatest variations in specific biovolumes were recorded for filamentous bacteria (coefficients of variation ranged from 16 to 109%). These variations were more pronounced in the oxygenated and microaerophilic strata (DOC ≈1.5 mg liter−1). Fluctuations in cell volume were high (coefficients of variation =12–80%) for coccal bacteria, whereas no marked fluctuations were found for the rod and vibrio bacteria (coefficients of variation =4–10%). Evidence of diel patterns of cell volume of filamentous bacteria is provided. These cells displayed their maximum size during the day until early night, indicating cell division was occurring at night. Homogeneous circadian patterns were not provided by specific volume variations of coccal, rod, and vibrio bacteria. Statistical relationships between bacterial specific biovolumes and the biotic and abiotic parameters considered are discussed.
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Although in a strict sense the term ‘phytoplankton biomass’ only refers to living algal material, in aquatic ecology the term has been associated with a variety of biological and biochemical procedures used to quantify the particulate matter suspended in natural waters. Relative merits of different ‘biomass’ characteristics have been studied in three Dutch freshwater lakes with great differences in absolute biomass. Parallel determinations have been made of seston dry weight and supplementary elementary and caloric analyses of seston, of chlorophyll-a concentration and supplementary paper chromatographic analyses of pigment extracts, of particle concentration and particle size distribution as studied with an electronic particle counter, and of phytoplankton cell volume as calculated from the results of microscopic enumeration and sizing of algae. In this way an attempt was made to create a detailed picture of the nature of the seston of the three freshwater lakes. Different analytical techniques give strikingly different information, the accuracy of any method is largely dependent on the circumstances present, and different biomass characteristics therefore are only of value in limited spheres. It is suggested to distinguish between total seston characteristics (e.g. seston dry weight, particulate organic carbon, total particle volume) and strictly algological biomass characteristics (e.g. chlorophyll-a concentration, phytoplankton cell volume). The pattern of growth of phytoplankton populations shown by e.g. chlorophyll-a concentration may differ markedly from that indicated by e.g. total particle volume or seston dry weight. Also, to more or less extent the wax and wane of phytoplankton populations may go undetected among the total seston. Apparently, there is no one method of estimating biomass and no conversion factor that may serve for general purposes. In general, unambiguous information on the nature of the seston of natural waters may only be obtained by estimating total seston characteristics and algological biomass characteristics simultaneously. Depending on the objective of the investigation supplementary component analyses should be carried out to guarantee the correct interpretation of the results.
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The relationship between measurements of phytoplankton cell volume (CV) and chlorophyll a (Chla) in the near surface waters (1 m) of 12 North Island lakes was investigated between January and December 1982 to determine the usefulness of Chla as a biomass indicator. A strong linear relationship was obtained between the two parameters (F = 262, n = 131) which explained 66% of the variation. There was a distinct seasonal trend, however, in the amount of chlorophyll a measured per unit algal cell volume (Chla/CV). Values of Chla/CV ranged between .1.9 and .12.4 μ and were generally high dunng the wlO.ter (interquartile range 6.6-10.5, n = 33) and low dunng the summer (interquartile range 2.8-3.9, n = 47). When this effect was taken into account, the explanatory power of the Chla-CV relationship increased to 78%. This relationship was not significantly affected by changes in phytopla~kton taxonomic composition (i.e., Bacillariophyceae Chlorophyceae, Dinophyceae), and reliably predicts either Chla or CV in other North Island lakes. Application of the predictive relation is discussed and further corroboration of the relation is encouraged.
The use of adensine triphosphate (ATP) analysis for freshwate algal and plankton populations was evaluated as a measure of biomass and as a bioassay response parameter. ATP analysis was performed using the firefly luminescence procedure. In short term laboratory studies. ATP levels in cultured algae and lake plankton correlate well with other standard biomass parameters, including chlorophyll a and dry weight. Algal ATP responded rapidly to mercury addition and pH changes, indicating its usefulness as a measurement of toxicity. The rapid response of ATP following nutrient additions to starved algal cultures suggests ATP may be useful as a tool in nutrient bioassay studies.
The plankton of nine Ontario lakes spanning several physiographic regions was sampled every two weeks during the ice-free period of 1981, and one lake was studied in the three previous years. Phytoplankton, zooplankton, and ciliated protozoa were sampled, counted and sized. The size data were converted to biomass estimates to yield quantitative comparisons of the relative allocation of biomass among different functional compartments. This is the first study to look simultaneously and quantitatively at the total plankton system of lakes (including ciliates, pbytoplankton and net zooplankton) over a broad physiographic region. Ciliates constitute -10% of the non-algal biomass and 5% of the total planktonic biomass of these lakes. Ciliate standing crops among lakes are significantly corrrelated with total organic and total inorganic carbon concentrations in the water column, while the dynamics of ciliate biomass fluctuations are significantly correlated with variations in total phosphorus concentration, in conductivity, in Kjeldahl nitrogen concentration, and in inorganic carbon content. There appears to be a significant dynamical relationship between ciliates as a proportion of the total planktonic biomass, exclusive of filamentous and large (>30 μm) spherical algae, and the relative biomass of small algae (2-5 μm) as a fraction of total algal biomass, again exclusive of filaments and large (>30 μm) algae. The hypothesis is advanced that ciliates primarily function as bacterial grazers in planktonic ecosystems and that their primary competitors in this role are rotifers.
Skeletonema costatum and Coccolithus huxleyi were grown in nitrogen-limited chemostat cultures with illumination provided in light/dark cycles. S. costatum assimilated nitrate and ammonium primarily during the day and less so at night. Conversely, the concentration of nitrate and ammonium in the culture medium varied periodically, increasing at night and decreasing in the light. C. huxleyi assimilated both N sources at a rate sufficient to keep them at very low levels both day and night. However, the activity of N-assimilating enzymes, measured in cell-free extracts, were higher in the light than in the dark periods, implying light/dark differences in the capacity to assimilate nitrogen. Such periodicity in the rate of uptake and enzymatic activity appears to complicate the mathematical expression of nutrient-limited growth of phytoplankton exposed to natural light/dark cycles. Three aspects of dial periodicity in N assimilation have been observed in natural phytoplankton communities in the sea: (1) in assimilation rate, (2) in activity of enzymes of N-assimilation, and (3) in the ammonium concentration of the seawater. The cultures also showed periodicity in these parameters and appear to be useful model systems for study.
ATP, carbon, and nitrogen content, and cell volume have been measured in 7 marine algae in culture. Intraspecific differences are negligible during the phase of exponential growth; interspecific differences in ATP and carbon content are slight during this phase compared with those observed in the same cultures between the exponential and senescent phases. As the interspecific differences agree well with those reported for algae in situ, this leads the authors to believe that the greater part of the biomass in situ is always in a state of physiological youth. ATP content is higher in diatoms, and seems linked with silica shell synthesis. In non-silicified species, there is a significant correlation between the ATP: plasma ratio and the division rate, although the cellular volumes are quite different. In the author's opinion, ATP content allows a good estimation of biomass to be made, as well as, under controlled conditions, a suitable estimation of primary productivity; however, because of their high silica content, diatoms should be considered separately from other phytoplankters.
An investigation into the spatial and temporal distribution of ciliate populations in a small monomictic eutrophic lake was carried out over a one-year period. A community of large cilates dominated by Loxodes striatus, L. magnus, Spirostomum teres and Frontonia leucas occurred in the deep water sediments during the period of winter mixis. These cilates vacated the sediment during summer stratification, progressively colonising the hypolimnion in phase with the upwards spread of anoxic conditions, returning once more to the sediment following destratification. The possible significance of this ciliate community is discussed.
Three methods for phytoplanktonic standing crop estimations have been used during ecological successions in a meso-oligotrophic lake (Le Pavin, France). In this study, it appears that cell counting and pigment concentrations overestimate the biomass of pioneer population, by taking into account a high number of dead cells. It seems preferable to estimate biomass from ATP concentrations, to measure living phytoplanktonic standing crop and to calculate activity coefficients. However, continuous rearrangements exist between ATP, ADP and AMP in the pool of adenine nucleotides. These rearrangements, which are revealed by energy charge variations, indicate changes in intracellular ATP concentrations. Consequently it is desirable to integrate energy charge variations in standing crop estimates from ATP concentrations.
Forrest, W. W. (University of Adelaide, Adelaide, South Australia). Adenosine triphosphate pool during the growth cycle in Streptococcus faecalis. J. Bacteriol. 90 1013–1018. 1965.—The adenosine triphosphate (ATP) pool has been studied throughout the growth cycle of Streptococcus faecalis. Normally, the pool is quantitatively related to the concentration of organisms and the growth rate, but deviations from these relationships can occur without affecting the growth rate of the organisms. A critical concentration of ATP seems necessary for exponential growth, and, at lower levels, only linear growth can occur. If no growth can take place, catabolism of added energy source gives rise to a large increase in the pool level. The pool level represents the balance between the demands of the organisms for energy and the supply of energy derived from catabolism of the substrate.
1.1. Three types of light-measuring devices have been used to measure the effect of various chemical agents on firefly luminescence. They are the commercially available Farrand photofluorometer, a recording DC IP21 photomultiplier, and a liquid-nitrogen-temperature quantum counter whose ultimate sensitivity is of the order of 10−9 g. ATP/ml.2.2. It has been possible to use both the crude and partially purified water extracts of Photinus pyralis to measure light production in the presence of “potential” or actual ATP by using arsenate buffer.3.3. Differential-determination of ATP and ADP in the presence of each other is possible through the use of myokinase.4.4. Phosphocreatine may be determined by the use of the appropriate transphosphorylase and adenylic acid.5.5. Glucose may be determined by the use of hexokinase and ATP, measuring the depression in luminescence.6.6. Chloride ion has a strong inhibitory effect on luminescence which must be controlled rigorously.7.7. Myokinase, hexokinase, apyrase, and creatine adenylic transphosphorylase may be determined in small concentrations with this system.8.8. Preliminary examination of certain biological processes with this tool suggests its wide applicability for routine surveys of phosphorylative energetic mechanisms.
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