An indirect method for the rapid estimation of carotenoid contents in Phaeodactylum tricornutum: Possible application to other marine algae

The equations of Parsons and Strickland (1963) have been applied to calculate the concentration of total carotenoids in Phaeodactylum tricornutum, and the same pigments have simultaneously been determined on the individual fractions after TLC (thinlayer chromatography) separation. Comparison of both sets of data shows that for diverse carotenoids: chlorophyll a ratios, the direct spectrophotometric values range from 80 to 130% of the chromatographic values; wide variations in the proportions of different carotenoids account for such difference. On the basis of correlations between the proportion of each carotenoid and the absorbance ratio 480:663 nm, an indirect method for the rapid estimation of individual carotenoids of P. tricornutum is proposed. Its eventual application to other Chrysophyta species and to sea water is discussed.Contribución del Instituto de Biología Marina No. 320.     

Microbial activity and accumulation of organic matter in the burrow of the mud shrimp, <Emphasis Type="Italic">Upogebia major</Emphasis> (Crustacea: Thalassinidea)

Microbial activity and accumulation of organic matter in the burrow of the thalassinidean mud shrimp, Upogebia major, were studied on a tidal flat along the northern coast of Tokyo Bay, Japan. The burrow of U. major is Y-shaped with an upper U-shaped part plus a lower I-shaped part. Its lower part can extend to a depth of 2 m. In the present study, we compare electron transport system activity (ETSA), bacterial abundance and organic matter content [total organic carbon (TOC), total nitrogen (TN) and chlorophyll a (chl. a)] of the burrow wall sediment with the tidal flat surface and non-burrow sediments. We also compared the U- and I-shaped part in terms of these parameters. ETSA in the burrow wall was higher than at the tidal flat surface in the warmer season, and was always higher than at surrounding non-burrow sediments. Bacterial abundance in the burrow wall was higher than at the tidal flat surface and surrounding sediment. TOC and TN contents in the burrow wall were two to three times higher than those at the tidal flat surface and non-burrow sediments, regardless of season. However, there was no significant difference in chl. a content between burrow wall and tidal flat surface. These results suggest that organic enrichment of the burrow wall is a result of organic matter particles such as phytodebris accumulation along the burrow wall. For all parameters of the burrow walls, no statistical differences were found between the two parts. Present results indicate that U. major actively transports the water containing suspended organic particles not only through the U-part but also into the deeper I-part. These indicate that burrow of the mud shrimp provides a dynamic environment for microbial community in tidal flat sediment.     

Sources and biochemical composition of suspended particulate material in a submarine cave with sulphur water springs

Sources, biochemical composition and nutritional value of suspended particulate material were investigated from February 1994 to February 1995 in a submarine cave (Grotta Azzurra, Capo Palinuro, southwestern Italy) with hot sulphur springs and associated mats of chemolithoautotrophic bacteria in its innermost dark part (Snow Hall). Concentrations of total suspended material (TSM), particulate inorganic material (PIM), organic carbon (POC), organic nitrogen (PON), chlorophyll a (Chl a), phaeopigments (Phaeo), carbohydrates (TCH), proteins (TPR) and lipids (TLI) were measured at four stations along an outside–inside transect, in order to address whether the quantity and quality of suspended particles varied over time with increasing distance from the entrance of the cave and estimate the relative contribution of chemosynthesis versus photosynthesis in supplying POC to cave heterotrophs. The abundance and biochemical composition of suspended material available to filter-feeders varied over time, but no significant quantitative or qualitative differences were detected along the outside–inside transect. Concentrations of TSM and of its different compounds (PIM, POC, PON, Phaeo, TCH, TPR, TLI) were homogeneous among the four stations or fluctuated without any consistent trend, with no apparent ageing and degradation of organic material in the innermost dark station. Conversely, concentrations of Chl a significantly decreased from outside to inside the cave at all sampling periods. It is suggested that suspended POC in Grotta Azzurra consists of mixed assemblages of particles coming from advection of photosynthetic material from the open sea and local inputs of carbon by sulphur-oxidizing bacteria. Based on POC/Chl a ratios, the relative contributions of chemosynthesis versus photosynthesis in supplying POC to benthic heterotrophs was estimated to be 31 and 69%, respectively. Received: 9 December 1996 / Accepted: 14 January 1997     

Spatial and Temporal Changes of Suspended Matter in Relation to Wind and Vegetation Cover in A Mediterranean Shallow Coastal Environment

Abstract

Seasonal and spatial changes in seston, (POC), particulate organic carbon, (PON) particulate organic nitrogen and chlorophyll-a concentrations were studied on a monthly basis in a Mediterranean shallow coastal area (Stagnone di Marsala, Western Sicily) in order to gather information on factors controlling particulate organic matter distribution and composition. Seston concentration and composition were connected to the main physicochemical and biological driving factors, such as temperature, salinity, dissolved oxygen, wind-speed and biomass of submerged vegetation. the Stagnone di Marsala is characterized by high temperatures with strong seasonality (range: 11–28°C), while values ranged from 33 to 45 salinity. Total suspended organic matter concentrations (by ignition loss) ranged from 2 mg l^?1 (in summer) to 12mgl^?1 (in winter) and chlorophyll-a concentrations from 0.02 to 2 μgl^?1. Despite a low POC/PON ratios (ranging from 5 to 11), the ratio of POC to chlorophyll (CHL-a) displayed very high values (annual average of 647). the data reported in this study, highlighting the oligotrophy of the Stagnone di Marsala area, indicate that the trophic state of the basin was controlled by different degrees of wind exposure (mean monthly wind velocity at exposed sites ranged between 4.2 and 6.7 m s^?1) and by gradients in vegetation cover. These two factors induced clear changes in the concentration and composition of the suspended particles, but played a different role in exposed and sheltered areas. Exposed areas with limited vegetation were characterized by large resuspension processes and wide temperature and salinity fluctuations caused by wind induced turbulence. in these areas, autotrophic biomass (as chlorophyll-a), due to phytoplankton and/or re-suspended microphytobenthos, appeared to play an important     

Particulate Organic Matter Composition in A Semi-Enclosed Marine System

Spatial and temporal changes in the biochemical composition of particulate organic matter in a semi-enclosed marine system (Stagnone di Marsala, Mediterranean Sea) were studied, on a monthly basis, from January to December 1994, in order to assess nutritional value of suspended particles for benthic suspension feeders. According to previous findings, the study site displayed a strong oligotrophy. Chlorophyll-a accounted for a very low fraction of the total suspended matter pool (0.1%), whereas at least 75% of POC was of detrital/heterotrophic origin. POC: PON ratio values indicate that bacterioplankton biomass accounted for a significant fraction of the total POC pool, displaying values comparable to those of the phytoplankton biomass (phytoplankton to bacterial biomass ratio was about (1). Temporal and spatial changes in the biochemical composition of particulate organic matter were rather limited and related to its sources, the main of which is represented by detrital particles released by the Posidonia oceanica (L.) beds. the comparison between our results and those encountered in other coastal lagoons indicates that the low abundance of suspension-feeding organisms observed in the study area is related to the “quality depression” of particles due to the dilution of high quality compounds (i.e., biopolymeric carbon) in a largely inorganic matrix. This result leads us to conclude that, to reach the same amount of high quality particulate food, a suspension feeder mollusc in the Marsala lagoon would need to filter a sea water volume around 3 times higher than in other Mediterranean coastal lagoons.     

Role of grapsid crabs, Parasesarma erythrodactyla, in entry of mangrove leaves into an estuarine food web: a mesocosm study

The influence of the crab Parasesarma erythrodactyla on the entry of the organic matter derived from Avicennia marina mangrove leaves in a sub-tropical mangrove ecosystem of southeast Queensland, Australia, was simulated using tidal mesocosms. Degradation of mangrove leaf organic matter was followed by analysing the fatty acid composition, carbon, and nitrogen isotopic signatures of the surface sediment and suspended particulate organic matter (SPOM) with and without the presence of crabs. Assimilation of mangrove organic matter by P. erythrodactyla was also assessed by stable isotope and fatty acid analyses in tissues and faeces. Results of the chemical tracer analyses question the adaptability of P. erythrodactyla to a diet comprised exclusively of mangrove leaves, and suggest that these organisms were dependent on additional food sources in their natural environment. Crab processing of senescent leaves significantly accelerated the transfer of mangrove organic matter to the surface sediments, as shown by a higher C/N ratio, a higher contribution of long-chain fatty acids and a more depleted C isotopic signature of sediment samples in the mesocosms with crabs compared to those without crabs.     

Origin and Distribution of Suspended Organic Matter As Inferred From Carbon Isotope Composition in A Mediterranean Semi-Enclosed Marine System

Abstract

The origin and distribution of suspended organic matter, the trophic features and the stable carbon isotopic composition of particulate organic carbon (POC) were studied monthly in a Western Mediterranean semi-enclosed basin. Sampling stations were selected as a function of wind-exposure and the degree of vegetation cover and then compared with an adjacent unvegetated site. the predominant vegetation was seagrass (Posidonia oceanica and Cymodocea nodosa) and Caulerpa prolifera. Water samples were analyzed for total suspended matter (inorganic and organic fractions), photosynthetic pigments (chlorophyll-a and phaeopigments), dissolved organic carbon, particulate organic carbon and their isotopic composition. Temperature and salinity were also measured at the same sampling sites within range of Mediterranean limits. the suspended organic matter concentration was 1.77 ± 1.55 mg l^?1; the chlorophyll-a concentration was low (0.35 ± 0.24 μg l^?1); the disolved organic carbon concentration was 2,140 ± 2,010 μg l^?1; the particulate organic carbon concentration was 212 ± 106 μg l^?1 and the isotopic composition was 18.77 ± 2.51%_°. There were significant temporal differences except for phaeopigments, POC and its POC isotopic composition, and there were no spatial differences other than for δ¹³C. This picture highlighted a general seasonal trend and trophical features similar to adjacent sea.

Spatial differences in δ¹³C showed that the source of suspended organic matter was different between stations as that between sources and wind-hydrodynamic constraints. In     

A new method for estimating phytoplankton growth rates and carbon biomass

A new method is described for the determination of phytoplankton growth rates and carbon biomass. This procedure is easy to apply and utilizes the labeling of chlorophyll a (chl a) with ¹⁴C. Pure chl a is isolated using two-way thin-layer chromatography, and the specific activity of chl a carbon is determined. Data from laboratory cultures indicate that the specific activity of chl a carbon becomes nearly equal to that of total phytoplankton carbon in incubations lasting 6 to 12 h and can be used to calculate phytoplankton growth rates and carbon biomass. Application of the method to the phytoplankton community in an eutrophic estuary in Hawaii indicates that the cells are growing with a doubling time of about 2 d and that about 85% of the particulate carbon consists of phytoplankton carbon.     

Gas-liquid chromatograms of sesquiterpenes as finger prints for soft-coral identification

A possible additional means for aiding in the identification of soft corals based on their sesquiterpene composition, as determined by gas-liquid chromatography (GLC), is discussed. The use of this method for several species of Sinularia and Sarcophyton is illustrated. Several sesquiterpenes were identified, some of them for the first time from marine origin. Preliminary tests indicate that the sesquiterpene composition in the tested soft corals remained quite constant during different seasons of the year. It is suggested that such finger prints are produced by the corals themselves and not by the zooxanthellae, and that they are species-specific.     

A report of green algal pigments in the Central North Pacific Ocean

Thin-layer chromatography showed that chlorophyll b was present in acetone extracts of 6 samples of suspended matter collected in February 1973 from the Central North Pacific Ocean. Pigment patterns showed the presence of green algae and diatoms through most of the euphotic zone down to 200 m. Ratios of chlorophyll b:a ranged from 0.2 to 0.5 at 5 and 100 m depths, but were less than 0.05 at 200 m.     

Ecological investigations on the zooplankton community in balsfjorden,northern Norway: Lipids and fatty acids in Meganyctiphanes norvegica,Thysanoessa raschi and T. inermis during mid-winter

Total lipid of Meganyctiphanes norvegica (M. Sars) contained 53% triacylglycerols and traces of wax esters, that of Thysanoessa raschi (M. Sars) contained 44% triacylglycerols and 10% wax esters and that of T. inermis (Krøyer) contained 28% triacylglycerols and 40% wax esters. The triacylglycerols of M. norvegica were relatively rich in 20:1 and 22:1 fatty acids and its traces of wax esters resembled those of calanoid copepods. The triacylglycerols of both Thysanoessa species were deficient in 20:1 and 22:1 fatty acids but were richer in 16:1(n-7) and 18:1 (n-7) acids than those of M. norvegica. The wax esters of T. raschi contained phytol as almost the only fatty alcohol and were rich in 16:0 and 18:1 (n-9) fatty acids. The wax esters of T. inermis contained mainly 16:0 and 14:0 fatty alcohols with lesser amounts of phytol and their dominant fatty acid was 18:1, especially the (n-9) isomer. The triacylglycerols of T. inermis had 18:4 (n-3) as the major polyunsaturated fatty acid. From these and other aspects of fatty acid and fatty alcohol analyses it is concluded that a major foodstuff of M. norvegica in Balsfjorden is wax ester-rich calanoid copepods. T. raschi and especially T. inermis are concluded to have much more preference for phytoplanktonic food. Results are discussed in terms of current knowledge of the lipid chemistry of krill in the northern and southern hemispheres.     

Production and biomass accumulation of periphytic diatoms growing on glass slides during a 1-year cycle in a subtropical estuarine environment (Bay of Paranaguá, southern Brazil)

Production rates, chlorophyll concentrations and general composition of periphytic diatom communities growing on glass slides were studied in relation to environmental parameters during one seasonal cycle in the Bay of Paranaguá, southern Brazil. Slides were routinely submersed at 1, 2 and 3 m depth and recovered weekly for microscopic examinations, analyses of chlorophyll, cell counts and in situ photosynthetic incubations using the Winkler titration method. Water samples were also collected at surface and bottom layers for determinations of temperature, salinity, nutrients and chlorophyll in the water. The periphytic community was mainly formed by epipelic and epipsammic species, dominated by Navicula phyllepta, Cylindrotheca closterium, Navicula spp. and Amphora sp. Weekly chlorophyll a and cell accumulations on slides varied from <1–32 mg m⁻² and up to 31 × 10⁸cells m⁻², respectively. Photosynthetic rates varied from <1 to 35 mg oxygen mg chlorophyll a ⁻¹ h⁻¹, with higher values in summer. Daily production varied from 5 to 3,600 mg oxygen m⁻² day⁻¹ (<0.01–1.4 g carbon m⁻² day⁻¹). Multiple regression analysis revealed that vertical differences in light conditions and grazing pressure jointly affected the influence of temperature on the seasonal patterns of cell densities and chlorophyll concentrations according to depth. Received: 27 April 2000 / Accepted: 16 August 2000     

Release of dissolved organic matter by some marine macrophytes

The amount of photoassimilated carbon released as dissolved organic carbon was investigated for 5 species of benthic marine macrophytes (Thalassia testudinum, Diplanthera wrightii, Acanthophora spicifera, Chondria dasyphylla, and Dictyota dichotoma) and 1 species of pelagic marine macrophyte (Sargassum natans). Release rates ranged between 0.006 and 0.053 mg C g^-1 h^-1. Percent release values ranged from 1.1 to 3.8%. Spermatophytes had slightly lower percent release values than algae. Investigations performed on the ability of release products to be utilized by heterotrophic organisms showed that 20 to 30% of the released organic carbon was assimilated within a 2-h period.     

Stable isotopic compositions of hydrothermal vent organisms

Stable isotopic analyses were used to study trophic relationships in two communities of deep-sea hydrothermal vent organism in the Pacific Ocean. The community at Hanging Gardens on the East Pacific Rise (21°N), sampled in 1985, is dominated by two species of vestimentiferan tubeworms; communities at Alice Springs and Snail Pits on the Marianas Back Arc Spreading Center (western Pacific), sampled in 1987, are dominated by gastropod mollusks, barnacles, and anemones. In both locations, carbon and nitrogen isotopic values of vent invertebrates are significantly different from those of non-vent invertebrates collected at 11°N on the East Pacific Rise and elsewhere in the deep-sea. These distinct isotopic compositions reflect local sources of organic carbon and nitrogen used by vent consumers. Many vent invertebrates lacking chemoautotrophic endosymbionts have ¹³C-enriched values of-11 to-16%. compared to values of-17 to-22%. normally observed in deep-sea fauna. This suggests that a ¹³C-enriched food source is trophically important in both vent communities. Free-living bacteria colonizing surfaces and suspended in the water column may constitute this food resource. Nitrogen isotopic analyses show that the food web of the East Pacific Rise community has more trophic levels than the Marianas vent community.     

Molecular phylogenies help resolve taxonomic confusion with Asian <Emphasis Type="Italic">Crassostrea</Emphasis> oyster species

Identification of oyster species is still largely based on phenotypic characters that are highly plastic. Prompted by the proposed introduction of the Asian oyster species Crassostrea ariakensis into the Chesapeake Bay region of the U.S.A., this study uses molecular genetic information to understand the taxonomic framework surrounding C. ariakensis and to confidently distinguish among various sympatric oyster species. Putative samples of C. ariakensis and other species of cupped oysters from across Asia were collected and phylogenetic analyses were conducted on DNA sequences of both nuclear (ITS-1) and mitochondrial (COI) regions. Trees generated based on the two independent molecular datasets were highly congruent, and indicate that many oysters collected for this study as C. ariakensis were originally misidentified. Results also indicate that C. ariakensis, C. hongkongensis and C. nippona are distinct, but closely related species. There is strong support in both analyses for a close relationship between C. gigas and C. sikamea, as well as between C. belcheri and C. gryphoides, and between C. iredalei and C. madrasensis. The parsimony analyses based on these DNA markers, however, did not provide evidence to support C. angulata as a distinct species from C. gigas. Overall, the results emphasize the need for rigorous species identification, and additional extensive and intensive sampling to more accurately determine relationships among Crassostrea species, define their geographic distributions, and establish existing sympatry patterns.     

Comparison between remote-sensed data and in situ measurements in coastal waters: The taranto sea case

Monitoring and managing small coastal ecosystems requires a considerable understanding of the temporal dynamics of biophysical factors describing the coastal water systems. For this reason, daily observation from space could be a very efficient tool. The objective of the work described in this paper is to evaluate the contribution of remote sensing to the continuous monitoring of coastal areas. It is well known that in coastal areas, the presence of inorganic suspended sediments and coloured dissolved organic matter can make chlorophyll-concentration measurements from remote sensing difficult. To overcome these difficulties, an alternative approach to the SeaWiFS standard chlorophyll algorithm is presented, based on a semi-analytic model for sea water and on the use of MODIS data as input in a model for atmospheric effects removal. Moreover, land contamination (mixed sea–land pixels) can introduce ambiguities in sea-surface temperature measurements from remote sensing. This paper proposes the use of a hydrodynamic model as a time–space interpolator of in situ campaign data, to extensively validate the temperature values extracted from AVHRR sensor. We validated the proposed approach, using experimental field data collected over a two-year campaign in the Taranto Gulf. The results seem to indicate a good agreement between remote-sensed and in situ data.     

Symbiosis in sacoglossan opisthobranchs: functional capacity of symbiotic chloroplasts

It has been previously reported that many species of the order Sacoglossa (Mollusca: Opisthobranchia) contain algal chloroplasts within the cells of their digestive gland and maintain them in a symbiotic condition. In the present study, two species, Elysia hedgpethi Marcus and Placobranchus ianthobapsus Gould, were compared as to their abilities to retain functional chloroplasts in their tissues. Animals were starved for varying lengths of time, and the functional capacity of the plastids was ascertained at intervals. The chlorophyll content of whole animals, and the ability to incorporate ¹⁴CO₂ were used as the assay for functional capacity. E. hedgpethi decreased in chlorophyll content during starvation until the tenth day, when no chlorophyll was detectable spectrophoto-metrically. Incorporation of ¹⁴CO₂ paralleled the decline in chlorophyll, and was at control levels by the tenth day. P. ianthobapsus showed no decline in chlorophyll content over 27 days starvation, although the ability to incorporate ¹⁴CO₂ showed a decrease.     

Mechanisms to initiate a Heterosigma akashiwo red tide in Osaka bay

The ecological role of diel vertical migration of Heterosigma akashiwo Hada to initiate red tide was investigated in Tanigawa Fishing Port and Sano Harbor, Osaka Bay, Japan during red tide seasons in 1979 and 1980. This species migrated toward the surface early in the morning at a velocity of 1.0 to 1.3 m h^-1. Downward migration was found in the afternoon, and more than 2.0×10³ cells ml^-1 aggregated in the bottom layer at night. The upward migration started before sunrise and downward shifting occurred prior to sunset. The movement is presumably associated with circadian rhythm, which is known as one of the biological periodicities. H. akashiwo crossed steep temperature and salinity gradients (6.5°C and 5.7 S) during the diel vertical migration. High values of particulate organic carbon and nitrogen concentrations were obtained in dialysis bags suspended in situ at identical layers with high cell concentration, while the values for surface and bottom bags were comparatively low. The results reveal that H. akashiwo migrates toward the sea surface to carry out photosynthesis effectively, and to the bottom to utilize nutrients efficiently.     

Characterization of in vivo absorption features of chlorophyte,phaeophyte and rhodophyte algal species

Despite the plentiful diversity of macroalgae in coastal environments, few studies have examined the in vivo absorption features of common marine macrophytes. Here we report on results of a survey of 12 central California common intertidal and subtidal taxa, representing Chlorophyta, Phaeophyta and Rhodophyta carried out in the summer of 1988. Computer-assisted analyses were used to obtain fourth-derivative spectra from in vivo absorption spectra determined at room temperature to obtain spectral diagnostics for the different algal divisions and to provide a means to determine whether spectral features could be used to identify stress responses among these plants. Among the Chlorophyta, characteristic maxima for chlorophylls a and b were resolved in all species examined, and a spectral component attributable to siphonaxanthin-like carotenoid(s) was observed in two Ulva species and the coenocytic alga Derbesia marina. Representatives of the Phaeophyta were characterized by similar maxima for chlorophyll a and c, and for fucoxanthin. Among the Rhodophyta, maxima for chlorophyll a, as well as B- and R-type phycoerythrin were resolved. Differences in in vivo absorption features were detected for two tidal populations of Porphyra perforata and Mastocarpus papillatus. High-tidal thalli absorb less green light in regions characterized by phycoerythrin, but have enhanced carotenoid absorption compared with lowtidal thalli. Resolution of spectra by fourth-derivative analysis revealed significant differences in phycoerythrin and carotenoid contents. The spectral changes observed appear to reflect environmental and possibly populational characteristics of these algae. The spectral analyses described here provide robust, non-invasive means to characterize subtle responses of macroalgae to environment in ways not possible previously. Additional merits of these fourth-derivative analyses for use in environmental studies are discussed.This contribution is dedicated to the memory of Professor L. R. Blinks, a pioneer in photosynthesis research and algal physiology     

Marine diatoms grown in chemostats under silicate or ammonium limitation. III. Cellular chemical composition and morphology of Chaetoceros debilis,Skeletonema costatum,and Thalassiosira gravida

Three marine diatoms, Skeletonema costatum, Chaetoceros debilis, and Thalassiosira gravida were grown under no limitation and ammonium or silicate limitation or starvation. Changes in cell morphology were documented with photomicrographs of ammonium and silicate-limited and non-limited cells, and correlated with observed changes in chemical composition. Cultures grown under silicate starvation or limitation showed an increase in particulate carbon, nitrogen and phosporus and chlorophyll a per unit cell volume compared to non-limited cells; particulate silica per cell volume decreased. Si-starved cells were different from Si-limited cells in that the former contained more particulate carbon and silica per cell volume. The most sensitive indicator of silicate limitation or starvation was the ratio C:Si, being 3 to 5 times higher than the values for non-limited cells. The ratios Si:chlorophyll a and S:P were lower and N:Si was higher than non-limited cells by a factor of 2 to 3. The other ratios, C:N, C:P, C:chlorophyll a, N:chlorophyll a, P:chlorophyll a and N:P were considered not to be sensitive indicators of silicate limitation or starvation. Chlorophyll a, and particulate nitrogen per unit cell volume decreased under ammonium limitation and starvation. NH₄-starved cells contained more chlorophyll a, carbon, nitrogen, silica, and phosphorus per cell volume than NH₄-limited cells. N:Si was the most sensitive ratio to ammonium limitation or starvation, being 2 to 3 times lower than non-limited cells. Si:chlorophyll a, P:chlorophyll a and N:P were less sensitive, while the ratios C:N, C:chlorophyll a, N:chlorophyll a, C:Si, C:P and Si:P were the least sensitive. Limited cells had less of the limiting nutrient per unit cell volume than starved cells and more of the non-limiting nutrients (i.e., silica and phosphorus for NH₄-limited cells). This suggests that nutrient-limited cells rather than nutrient-starved cells should be used along with non-limited cells to measure the full range of potential change in cellular chemical composition for one species under nutrient limitation.Contribution No. 943 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA.