Lipid Metabolism in the Brown Marine Algae Fucus vesiculosus and Ascophyllum nodosum

Lipid metabolism and environmental effects on this process havebeen studied in the marine brown algae Fucus vesiculosus andAscophyllum nodosum. These algae showed very similar patternsof lipid metabolism during 24 h incubations. Labelling from[1-¹⁴C]acetate showed the major labelled lipids to be the ß-alanineether lipid and the neutral lipid fraction in both algae. Ofthe glycolipids, only sulphoquinovosyldiacylglycerol was welllabelled and the phosphoglycerides were all poorly labelled.The major labelled fatty acids were palmitate and oleate, againin both algae, although Fucus vesiculosus also showed significantlabelling of stearate and behenate. Although the amount of fattyacid labelling increased with time, the proportion of labelin palmitate and oleate remained approximately constant. Verylong chain fatty acids (arachidic, behenic) were increasinglylabelled with time. Lowered incubation temperatures decreased labelling of the saturatedfatty acids. Cu²⁺ increased the proportion of oleate labelledin both algae, and of linoleate in Fucus vesiculosus. This cationdecreased the percentage labelling of stearate and myristatein Ascophyllum nodosum. Lipid metabolism in Ascophyllum nodosumwas more sensitive to raised Cu²⁺ levels than in Fucus vesiculosus Key words: Acyl lipid metabolism, Fucus vesiculosus, temperature effects, Ascophyllum nodosum, copper pollution     

COMPARATIVE STUDIES OF PHOTOCHEMICAL OXIDATION-REDUCTION REACTIONS IN LAMELLAR FRAGMENTS OF VARIOUS ALGAE AND SPINACH

The activity of various electron carriers, including DPIP, spinachplastocyanin, mammalian cytochrome c, and Anabaena cytochrome553, as donor in the reaction induced by the photochemical systemI was examined with lamellar fragments of various algae andspinach. Reduced DPIP was an effective electron donor irrespective ofthe organisms, when it was supplied at a high concentration(10^–3 M). Spinach plastocyanin was effective in the reactionswith the lamellae of green algae, Euglena, diatom Phaeodactyrumand red algae Porphyra yezoensis and Porphyra sp. Yamamoto II,whereas it was inactive in the lamellae of blue-green algae.Horse-heart cytochrome c and Anabaena cytochrome 553 were activein the reaction with the lamellae of bluegreen algae. The formercytochrome was also active in the reactions in Porphyridiumand Cyanidium. The cytochromes were less active in the reactionsin which spinach plastocyanin acted as effective electron donor. The data were interpreted as that the photochemical system Iin bluegreen algae differs from that of other photosyntheticorganisms with respect to the properties of the site of theelectron-input. ¹ Present address: Nomura Research Institute for Technologyand Economics, Kamakura, Kanagawa. ² Present address: Ocean Research Institute, University of Tokyo,Nakano, Tokyo.     

The Lipid Phase of Thylakoid and Cytoplasmic Membranes from the Blue-Green Algae (Cyanobacteria), Anacystis nidulans and Anabaena variabilis

The lipid phases of the thylakoid and cytoplasmic membranesfrom the blue-green alga, Anacystis nidulans, were studied bya spin-probe method using 2-(14-carboxytetradecyl)-2-ethyl-4,4-dimethyl-3-oxazolidinyloxyl.The thylakoid and cytoplasmic membranes of this alga were bothin the liquid crystalline state at growth temperature, and inthe phase separation state at about 0?C. The thylakoid membranesentered the phase separation state at a temperature higher thanthe cytoplasmic membranes. The lipid phase of the thylakoidmembranes from Anabaena variabilis was studied in a similarway, and these membranes were found also to undergo the phasetransition. The temperature for the onset of the phase separationand the fluidity of the membrane lipids of both algae dependedon the growth temperature of the culture. (Received April 9, 1984; Accepted June 1, 1984)     

Effect of temperature on growth and ingestion rates of Favella sp

This study describes the effect of temperature on the growthand ingestion rates of the tintinnid, Favella sp. cultured withthe dinoflagellate Heterocapsa triquetra. In vivo fluorescencewas used to monitor the change in density of the H. triquetrapopulation over 4- to 5-day periods in control tubes containingonly algae, and in experimental tubes containing algae and tintinnids.A ‘switchover point’ occurred in the temperaturedependency of the growth rate such that below 11.4°C, H.triquetra grew more quickly than Favella sp. and above thistemperature the situation was reversed. Ingestion rates of Favellaon H. triquetra were found to be temperature dependent in anonlinear fashion. The rate doubled (from 2.5 to 5.3 cells animal^–1h^–1) between 11.4 and 16.4°C whereas there was nochange in ingestion rates between 8.0 and 11.4°C, or between16.4 and 21.1°C.     

Specific growth rates of heterotrophic plankton organisms in a eutrophic lake during a spring bloom

The in situ growth of the dominating pelagic organisms at severaltrophic levels was investigated during a spring bloom characterizedby well-mixed cold water. The study includes primary productionand the carbon flow through the nano-, micro- and mesozooplanktonpopulations based on population dynamics and specific growthrates. The phytoplankton biomass and production were totallydominated by small algae <20 µm. of which {small tilde}5%were <3µm. potentially a food source for the nano-and microzooplankton. The mean carbon-specific primary productionwas 0.15 day^–1 and was regulated solely by light. Themean volume-based specific growth rate of bacterioplankton wasmodest. 0.1 day^–1. and probably controlled by the lowtemperature. The volume-based specific growth rates of heterotrophicnanoflagellates. ciliates. rotifers and copepods were 0.35.0.13. 0.16 and 0.03 day^–1, respectively. The observedgrowth of the heterotrophic plankton was generally not foodlimited, but was controlled by temperature. The stable temperatureduring the experiment therefore allows a cross-taxonomic comparisonof specific growth rates. The b exponent in the allometric relationship(G = aV^th) between volume-specific growth rate (G) and individualbody size (V) was –0.15 ± 0.03 for all filtratingzooplankton. indicating an in situ scaling not far from thephysiological principles onginally demonstrated for laboratorypopulations.     

Lipid Metabolism in Fucus serratus as Modified by Environmental Factors

The effect of changes in the environment on lipid metabolismhas been studied in the brown alga, Fucus serratus L. Lightstimulated the incorporation of radioactivity from /{^I4C/}acetateinto oleic and, especially, into linoleic acid. The same effectwas caused by lowering the incubation temperature from 15 °Cto 4 °C. Incubations in the presence of Cd ^{+ +}, Pb ^{+ +} orZn^{+ +} had no effect on the total uptake of /{¹⁴C/}acetate intothe frond tip samples, but lowered the labelling of total lipidsrelative to aqueous-soluble components. However, pre-exposureof the algae to heavy metal cations caused changes in the uptakeof radioactivity but had less effect on the relative labellingof lipids than incubations in the presence of heavy metal cations.Algae collected from sites where the dissolved levels of heavymetals were elevated, showed a decrease in the relative labellingof lipids from /{¹⁴C/}acetate. Concentrations of Cd^{+ +}, Pb⁺⁺ or Zn^{+ +} at 10 x levels found at the collection site had littleeffect on the pattern of fatty acids made by Fucus serratus. Key words: Lipid metabolism, Fucus serratus L., Environmental changes     

Enzymes involved in the last steps of the biosynthesis of mannitol in brown algae

Mannitol-1-phosphate dehydrogenase (EC 1.1.1.17 [EC] ) and mannitol-1-phosphatase(EC number yet unassigned) were detected in the brown algae,Spatoglossum pacificum and Dictyota dichotoma. The enzymes wereextracted from algal fronds and their properties were investigatedusing partially purified preparations. Mannitol-1-phosphatase shows maximum activity at pH 7. The enzymehad a narrow substrate specificity. The Km value for mannitol-1-phosphateis 8.3x10^–4 M (30°C, pH 7.0). The enzyme is activatedby Mg⁺⁺ and Mn⁺⁺and is strongly inhibited by PCMB, Hg⁺⁺and NaF. Mannitol-1-phosphate dehydrogenase showed maximum activitiesat pH values 6.5 and 10.2 in reductive and oxidative reactions,respectively. The dehydrogenase also showed narrow substratespecificity; mannitol-1-phosphate and NAD or fructose-6-phosphateand NADH₂ are utilized, respectively, in oxidative and reductivereactions by the enzyme. Km values for these substrates andthe coenzymes are 2.5x10^–4 M and 7.1x10^–5 M forthe first pair and 2.8x10^–4 M and 1.3x10^–5 M forthe latter pair. This enzyme was strongly inhibited by PCMBand Hg⁺⁺, but was only slightly affected by adenosine phosphates. Possible roles of these enzymes in the biosynthesis of mannitolin brown algae are discussed. ¹ Contributions from the Shimoda Marine Biological Station ofTokyo Kyoiku University, No. 233. This work was supported inpart by a Grant-in-Aid for Co-operative Research from the Ministryof Education, Japan and in part by a grant to one of us (T.Ikawa) from the Matsunaga Science Foundation. ² Present address: Chemical and Physical Laboratory, HoechstJapan Research Laboratory, Minamidai, Kawagoe, Japan. (Received February 22, 1972; )     

The effect of temperature on the post-embryonic growth of Neomysis intermedia Czernlawsky (Crustacea, Mysidacea) under laboratory conditions

The effect of temperature on post-embryonic growth of Neomysisintermedia was investigated under unlimited food conditionsin the laboratory. The effect of temperature on the size ofnewly released animals was negligibly small, but body size wasinversely related to temperature in adults. This was mainlycaused by the difference in the number of molts before maturation.The specific growth rate of N. intermedia increased exponentiallywith a temperature coefficient, Q₁₀ of 4.6 from 0.018 d^–13C to 0.21 d^–1 at 20C in juveniles, and with a temperaturecoefficient of 2.7 from 0.006 d^–1 at 3C to 0.05 d^–1at 25C in adults. The rate in juveniles levelled off above20C, and dropped at 29C. Brood size and brood interval decreasedwith temperature increase, while the daily specific reproductionrate increased. The specific growth rate of gravid females,including production of egg matter, increased exponentiallywith a temperature coefficient of 3.3 from 0.015 d^–1 at10C to 0.093 d^–1 at 25C. The present laboratory experiments confirmed the temperaturecontrol on the growth of N. intermedia suggested in a hyper-eutrophiclake.     

Photoacclimation in the marine alga Nannochloropsis sp. (Eustigmatophyte): a kinetic study

Exponentially growing cultures of Nannochloropsis sp. were transferredfrom high light (650 µ.mol quanta m^–2 s^–1)to low light (30 (µrnol quanta m^–2 s^–1). Thekinetics of changes in the various parameters of the photoacclimationprocess were followed until steady state was reached. In thisorganism, the observed 4.25 increase in cellular chlorophylla in the low-light-acclimated cells was linearly correlatedwith an increase in the number of photosynthetic units (PSU),while the PSU size remained constant. The new steady state wasreached after a transition period of 87 h and following an initialovershoot in the chlorophyll a concentration. During the processof photoacclimation, harvested and utilized light was increaseddue to a combination of increased cellular chlorophyll and anincrease of the quantum yield. These improvements in efficiencywere coupled with the reduction of respiration. The overalleffect of photoacclimation, therefore, amounted to reducingthe impact of low light on the growth rate of the algae. Ourstudy demonstrates the importance of the photoacclimation processin the context of the planktonic way of life. An index of the'effectiveness of photoacclimation' is proposed.     

Temperature dependence of chlorophyll {alpha} fluorescence in lettuce and spinach chloroplasts at sub-zero temperatures

The temperature dependence of chlorophyll fluorescence wasmeasured in spinach and lettuce chloroplasts at sub-zero temperaturesin the presence of 50% ethylene glycol. In the presence of 5mM Mg²⁺, a fluorescence maximum appeared at –31?C in boththe spinach and lettuce chloroplasts, while in the presenceof only 5 mM Na⁺ as cations the maximum shifted to –20?Cin the spinach chloroplasts and to –11?C in the lettucechloroplasts. Since the occurrence of a maximum in the temperatureversus fluorescence curve is an indication for the transitionof the physical phase of thylakoid membrane lipids between theliquid crystalline and the phase-separation state (16, 18),these findings suggest that the (major) phase transition ofmembrane lipids occurs at these low temperatures in chloroplastsof higher plants and also that the phase transition temperatureis markedly lowered by the presence of divalent cations. Ethylene glycol at a concentration of 50% had almost no effecton the temperature dependence of chlorophyll fluorescence ina lamellar membrane preparation of Anabaena variabilis. In awater suspension of dimyristoylphosphatidylcholine, the additionof ethylene glycol to 50% did not alter the characteristic featureof the temperature dependence of fluorescence of 1-anilinonaphthalene-8-sulfonate.These findings suggest that 50% ethylene glycol does not affectthe temperature of the transition of the physical phase of membranelipids. ¹ C.I.W.-D.P.B. Publication No. 592. ² Present Address: Department of Biophysics and Biochemistry,Faculty of Science, University of Tokyo, Hongo 113, Tokyo, Japan. (Received June 22, 1977; )     

Steady-state rotifer growth in a two-stage, computer-controlled turbidostat

Steady-state populations of rotifers (Brachionus calyciflorus)were maintained in twostage, continuous-flow turbidostatic cultureon the green alga Chlorella pyrenoidosa. In this system, themaximum specific growth rate,µ^max of the rotifers wasmaintained by using a computer to control the concentrationof algae, as rotifer food, in the rotifer culture. As rotifersconsumed algae, the turbidity decreased until a set-point wasreached. Then fresh algal suspension (supplied from a steady-statealgal chemostat) was metered into the rotifer culture, whichwas held in the dark. Rotifer and algal populations, as wellas rotifer µ^max entered steady states. These steady-stateresults were consistent with previous data from chemostat studies,but growth transients indicated that the of the µ^maxrotifersmay be subject to selection. The system is unique in providinga means to explore population dynamics of a metazoan maintainednear its µ^max.     

Respiration losses in planktonic green algae cultivated in raceway ponds

Rates of respiratory oxygen uptake were measured in darkenedsamples of green microalgae (Coelastrum sphaericum, Scenedesmusfalcatus) growing in raceway ponds under field conditions andindoors. The rates measured over 6–26h varied between1 and 1.5 and 8.25 µl O₂ mg^–1 h^–1 and dependedon incubation, temperature, time in the dark, and on the temperatureand irradiance at which the algae were cultivated before. Q₁₀values ranged from 1.43 to 1.68. Arrhenius relationships describedthe influence of temperature upon respiration rates below theoptimum temperature, even after many hours of incubation inthe dark. Respiration rates were lower after growth at optimaltemperature than after cultivation at other temperatures. Thelight history also influenced the rates, being high when thealgae were darkened after exposure to high irradiance and lowerafter weaker pre-illumination. For the algae under study theoverall loss during 12h of darkness was estimated to be 2–10%of the biomass prior to darkening. Higher losses are expectedfor natural conditions and stronger irradiances.     

Light, temperature and nitrogen as interacting factors affecting diel vertical migrations of dinoflagellates in culture

Diel vertical migrations of the marine dinoflagellates Gonyaulaxpolyedra Stein and Ceratium furca (Ehr.) Clap, et Lachm. werefollowed in a laboratory tube (2.02 m x 0.25 m) under a 12:12hlight:dark cycle. The effects of temperature stratification,two levels of surface irradiance and nitrogen depletion on patternsof vertical migrations were examined. At temperatures between22–26°C with small temperature gradients, both speciesmigrated at a rate of 0.7 –1.0 m h^–1. Steeper thermoclines(ca. 0.8°C 0.1 m^–1) with temperatures below ca. 20°Ccaused a marked decrease in swimming speed which resulted inaccumulations of cells in these thermocline regions. Under conditionsof nutrient sufficiency both algae migrated into the surfacelayers at irradiance values of over 1000 µE m^–2s^–1. Increasing nitrogen depletion caused the downwardmigration of both algae to commence progressively earlier inthe day and before the end of the light period. The earlierdownward migrations enabled a more complete descent throughthe thermocline. Nitrogen depleted cells of Gonyaulax continuedto undertake vertical migrations but avoided high irradiancesthus forming subsurface maxima at irradiance levels close to150 µE m^–2 s^–1. Ceratium cells which exhaustedboth inorganic nitrogen and phosphorus ceased to migrate accompaniedby a large change in cellular fluorescence.     

The relative importance of food and temperature to copepod egg production and somatic growth in the southern Benguela upwelling system

The fecundity and somatic growth rates of Calanus agulhensisand Calanoides carinatus, the dominant large calanoid copepodsin the southern Benguela upwelling system, as well as the fecundityof several other common copepods, were measured between Septemberand March of 1993/94 and 1994/95. Mean egg production of mostcopepods was low at >30 eggs female^-1 day^-1 {Calanoides carinatus23.7, Calanus agulhensis 19.0, Neocalanus tonsus 16.1 and Rhincalanusnasutus 26.1), whereas the mean fecundity of Centropages brachiatuswas significantly greater (83.6 eggs female^–1 day^-1).This study also presents the first comprehensive field estimatesof the fecundity of Nanno-calanus minor (mean: 26.1 eggs female^–1day^–1, range: 0.0–96.2 eggs female^–1 day^–1)and of somatic growth of N6 and all copepodite stages of Calanoidescarinatus (decreasing from 0.58 day^–1 for N6 to 0.04 day^–1for C5). Somatic growth rates of Calanus agulhensis also declinedwith age: from 0.57 day1 for N6 to 0.09 day1 for C5. Data ongrowth rates were used to assess the relative importance offood [as measured by total chlorophyll (Chi) a concentration],phytoplankton cell size (proportion of cells >10 µm)and temperature to the growth of copepods. Multiple regressionresults suggested that fecundity and somatic growth rates werepositively related to both Chi a concentration and phytoplanktoncell size, but not to temperature. Although it was not possibleto separate the effects of Chi a concentration and phytoplanktoncell size, data from previous laboratory experiments suggestthat copepod growth is not limited by small cells per se, butby the low Chi a concentrations that are associated with theseparticles in the field. Despite growth not being directly relatedto temperature, a dome-shaped relationship was evident in somespecies, with slower growth rates at cool (<13°C) andwarm (>18°C) temperatures. The shape of this relationshipmirrors that of Chi a versus temperature, where poor Chi a concentrationsare associated with cool and warm temperatures. It is concludedthat the effect of food limitation on growth of copepods outweighsthat of temperature in the southern Benguela region. Sourcesof variability in relationships between growth and Chi a concentrationare discussed.     

Relative impacts of copepods, cladocerans and nutrients on the microbial food web of a mesotrophic lake

Calanoid copepods, rather than cladocerans, frequently dominatethe zooplankton of lakes in New Zealand. The potential consequencesof this domination for the microbial community of mesotrophicLake Mahinerangi, New Zealand, were determined by field experimentsin which Boeckella and Daphnia were added to in situ enclosuresin the presence and absence of added nutrients. Boeckella hamataat ambient densities (2 and 81^–1) rapidly and severelysuppressed ciliate population growth over 4 days, even whenmicrobial growth was enhanced by added nutrients, but effectsof copepods on other components of the microbial community (bacteria,photosynthetic picoplankton, heterotrophic nano-flagellates,algae) were slight. In contrast, Daphnia carinata at the samedensities (but 3-fold higher biomasses per litre) had a relativelyweak effect on ciliates, suppressing ciliate abundance onlyafter 4 days at 8 Daphnia 1^–1 (330 µg 1^–1);this daphniid density also depressed abundances of large bacterialrods, some photosynthetic picoplankton and the dominant alga,Cyclotella. These results highlight the relative importanceof specific trophic linkages in a microbial food web; they alsosuggest that the dominance of Boeckella in many southern hemispherelakes may account for relatively low ciliate abundances in theselakes.     

Purification and Characterization of a Ca2+-Dependent Protein Kinase from the Halotolerant Green Alga Dunaliella tertiolecta

A Ca²⁺-dependent protein kinase (CDPK) that has been partiallypurified and characterized previously [Yuasa and Muto (1992)Arch. Biochem. Biophys. 296: 175] was further purified to about20,000-fold from the soluble fraction of Dunaliella tertiolecta.The enzyme preparation contained 60- and 52-kDa polypeptidesboth of which phosphorylated casein as a substrate. Both polypeptidesshowed a Ca²⁺-dependent increase in mobility during SDS-PAGEand ⁴⁵Ca²⁺-binding activity after SDS-PAGE and electroblottingonto a nitrocellulose membrane, suggesting that both the 60-and 52-kDa CDPKs directly bind Ca²⁺. The protein kinase inhibitors,K-252a and staurosporine, inhibited the CDPK competitively withrespect to ATP. An antibody raised against the 60-kDa CDPK crossreactedwith both the 60- and 52-kDa polypeptides. Both molecular specieswere autophosphorylated in the presence of Ca²⁺, and a highlyphosphorylated 80-kDa band appeared in addition to these phosphorylatedbands at 60 and 52 kDa in SDS-PAGE. However, the specific activityof CDPK was not changed by prior autophosphorylation when theautophosphorylated enzyme was assayed as a mixture of thesephosphorylated molecular species. Only the 60-kDa polypeptidewas immunodetected in subcellular fractions of Dunaliella cells.The 52-kDa polypeptide increased during storage of the enzyme.These results suggest that the 52-kDa polypeptide is a proteolyticartifact produced during purification. Immunoreactive bandsof 60-kDa were detected in extracts of several green algae butnot in extracts of higher plants or a brown alga. ¹This research was partly supported by Grants-in-Aid from theMinistry of Education, Science and Culture, Japan (No. 06454013and 06304023) and Research Fellowship of the Japan Society forthe Promotion of Science for Young Sciencists. ²Research Fellow (PD) of the Japan Society for the Promotionof Science.     

Food selection and growth of the planktonic ciliate Coleps hirtus isolated from a monomictic subtropical lake

A strain of Coleps hirtus (Ciliophora, Prorodontida) was isolatedfrom the epilimnion of monomictic Lake Kinneret. Growth of thisciliate was tested in response to 12 species of planktonic algaeand seven species of cultured bacteria from lake isolates whichwere offered as food. Eight species of algae (one Cryptophyceaeand seven Chlorophyceae) and four bacteria supported good toexcellent growth of C.hirtus. Growth rates (µ) and doublingtimes (DT) ranged from 0.008 to 0.029 h^–1 and from 23.9to 90.8 h respectively. C.hirtus was able to grow on bacteriaat concentration levels as low as 2–8 x 10⁵ cells ml^–1.No correlation was observed between growth rate of C.hirtusand cell volume of the prey. ^aPresent address: Istituto di Ecologia, Universita di Parma,43100 Parma, Italy     

In situ determinations of bacterial selectivity and filtration rates by five cladoceran zooplankters in a hypertrophic subtropical reservoir

Cladoceran in situ feeding rates on natural bacteria labelledwith [methyl-³H] were studied in parallel with feeding ratedeterminations on ¹⁴C-labelled Chlorella in a hypertrophic subtropicalreservoir (Lake Hartbeespoort) through spring and summer (1986/87).Community filtration rates (CFR5) on bacteria and algae weresimilar, but selection for Chlorella (relative to natural bacteria)increased from midsummer in association with declining bacterialdensity and increasing dominance of ‘inedible’ componentsof the natural phytoplankton. Species-specific filtration rates(SSFRs) were determined for Daphnia pulexllongispina, Ceriodaphniareticulata, Diaphanosoma excisum, Bosmina longirostris and Moinamicrura during their respective seasonal occurrence in the studyperiod. SSFRs on algae and bacteria increased with body length(L, mm) in all species apart from Bosmina. Species-specificdifferences in absolute feeding rate (FR, ml animal^–1day^–1), the slope of the FR-L relationship and bacteriaselectivity were evident. The feeding rate of all cladoceranson bacteria is described by the power equation FR 5.231L^1.42FR values on bacteria relative to FR values on algae averaged     

Firefly assay of adenine nucleotides from algae: Comparison of extraction methods

Pools of ATP, ADP and AMP in algae were compared after extractionwith Trisbuffer, ethanol, chloroform, sulfuric acid, trichloroaceticacid and perchloric acid. Experimental organisms were Anacystisnidulans, Euglena gracilis, Chlorella kessleri and Scenedesmusobtusiusculus. Adenine nucleotides were determined by fireflyluciferase assay. Extraction of illuminated algae in situ with the different methodsresulted in great variations in the proportions between ATP,ADP and AMP. The sum of adenine nucleotides varied less. Extractionwith trichloroacetic acid or perchloric acid generally yieldedthe highest energy charge and ATP/ADP ratio. Experiments withilluminated Anacystis and Scenedesmus showed that extractionwith perchloric acid in situ gave higher yields of ATP in theextracts than if samples were withdrawn for extraction (timefor sample withdrawal {small tilde}1 sec). For Anacystis, itwas demonstrated that the extraction methods giving the highestproportions of ATP were suitable for studies on fluctuationsof the adenine nucleotide pools at transitions from dark tolight. ¹ Permanent address: National Defence Research Institute, Department4, S-172 04 Sundbyberg, Sweden. (Received September 4, 1978; )     

Ammonium release by zooplankton in suspensions of heat-killed algae and an evaluation of the flow-cell method

The maximum excretion rate of NH₄ (39 nmol mg dry wt^–1h^–1) was directly measured for Daphnia pulex by measuringNH₄ accumulation in bottles containing D. pulex and dense, satiatingsuspensions of heat-killed algae. Ammonium release rates inthe algal suspensions were compared to those of individual animalsremoved from the suspension and placed in flow cells. Ammoniumrelease rate, R (nmol mg dry wt^–1 h^–1). in the flowcell decreased very rapidly with time, t (min), after removalaccording to the relation R = 26 + 25e^–0.16t. Ammoniumexcretion obtained by the flow cell method after extrapolationto time zero was not significantly different from that obtainedin the bottles. The considerable experiment-to-experiment variationin NH₄ excretion was in large part correlated (r² = 0.73) withthe feeding rate on the algae.