Optimal assay conditions for aspartate transcarbamylase (ATCase) activity in mesozooplankton

The optimal conditions for storing and assaying mesozooplanktonaspartate transcarb amylase (ATCase) (EC 2.1.3.1 [EC] .) are definedin order to enable the specific activity of this enzyme to beused as an index of mesozooplankton productivity. ATCase activitywas found to be stable for more than 2 weeks when stored inliquid nitrogen, with a negligible loss of activity, whateverthe state of frozen material (cell-free homogenate or wholecells). As an alternative, –20C, –90C freezersand dry ice (–80C) can give more flexibility for storingand transporting material, provided initial freezing has beencarried out at –196C. ATCase specific activity was stableover a large range of total protein concentrations (2–55mg ml^–1) for most mesozooplankton samples, and the enzymeactivity was linear between 0.8 and 80 nmol carbamyl aspartatemin^–1 mg^–1 of total protein. The optimal incubationtime (30 min), temperature (35C) and pH range (9–9.5)for assay of ATCase were similar forCalanus helgolondicus (Crustacea:Copepoda) (G.O.Sars) and three mesozooplankton assemblages sampledoff Plymouth (UK) in May, August and November. However, theKm values for both substrateswere variable: from 3.9 to 79.1for aspartate, and from 0.83 to 3.46 for carbamyl phosphate.Such variations arelikely to be due to the presence of differentATCase catalytic potential, rather than changes in ATCase assemblage.     

Egg production and hatching success in the calanoid copepods Calanus helgolandicus and Calanus finmarchicus in the North Sea from March to September 2001

Spatial and seasonal egg production rates (E_r) and egg hatchingsuccess in the copepods Calanus finmarchicus and Calanus helgolandicuswere measured in the North Sea from March to September. Foodavailability was monitored by chlorophyll and protist concentrationsand three size fractions of seston fatty acids. Seasonal andspatial distribution and production differed between the species.Calanus finmarchicus was found only offshore of the 50-m isobath,with decreasing E_r (37–28 eggs female^–1 day^–1)from March to July. Calanus helgolandicus had two abundancepeaks, in spring and autumn, with a low in May during whichtime the highest E_r were observed (38 eggs female^–1 day^–1).At other times, E_r in C. helgolandicus remained lower than inC. finmarchicus (     

Temporal variability in the night-time distribution of epipelagic siphonophores in the North Atlantic Ocean at Bermuda

The temporal variability of siphonophores was compared on night-to-night,month-to-month and year-to-year scales in open-net collectionsmade at or near the Bermuda Atlantic Time Series (BATS) site.These collections were made as oblique tows to 100 m on (i)one summer night in each of 1986, 1989 and 1991, (ii) one nightper month on 13 cruises between April 1991 and August 1992,and (iii) three consecutive summer nights of 1992. The meanwet displacement volume (WDV) of sipho nophores in the thesecollections was 1.4 ± 0.6 ml per 100 m³ or 20 ±10% of the total catch biovolume. The most common species belongedto the Calycophorae families Diphyidae and Abylidae: the combinednumerical abundance of Chelophyes appendiculala, Eudoxoidesmitra, Lensia subtilis. Eudox-oides spiralis, Bassia bassensisand Abytopsis eschscholtzi averaged 300–400 per 100 m³.Combined numbers of these common species varied little whensampled hour by hour at the same location during consecutivesummer nights or at different night-time sampling locations10¹–10² km apart when these were sampled within 3–5h. In contrast, most of these six species exhibited 2- to 3-folddifferences in night-time abundance between summers of differentyears and between seasons. Three seasonal groups were detected,with a winter group being the most distinct from the others.Changes in the abundance of several species were correlatedwith seasonal changes in the depth of the mixed layer.     

Copepod abundance in the western Irish Sea: relationship to physical regime, phytoplankton production and standing stock

The distinct patterns of stratification in the North Channeland stratified region of the western Irish Sea influence theseasonal abundance of phytoplankton. The 3–4 month productionseason in the stratified region was characterized by productionand biomass peaks in the spring (up to 2378 mg C m² day^–1and 178.4 mg chlorophyll m^–2) and autumn (up to 1280 mgC m^–2 day^–1 and 101.9 mg chlorophyll m^–2).Phytoplankton in the North Channel exhibited a short, late productionseason with a single summer (June/July) peak in production (4483mg Cm^–2 day^–1) and biomass (–160.6 mg chlorophyllm^–2). These differences have little influence on copepoddynamics. Both regions supported recurrent annual cycles ofcopepod abundance with similar seasonal maxima (182.8–241.810³ind. m^–2) and dominant species (Pseudocalanus elongatusand Acartia clausi). Specific rates of population increase inthe spring were 0.071 and 0.048 day¹ for the North Channel andstratified region, respectively. Increased copepod abundancein the stratified region coincided with the spring bloom, andwas significantly correlated with chlorophyll standing stock.Increased copepod abundance preceded the summer production peakin the North Channel. This increase was not correlated withchlorophyll standing crop, suggesting that a food resource otherthan phytoplankton may be responsible for the onset of copepodproduction prior to the spring bloom. Hetero-trophic microplanktonas an alternative food source, and advection of copepods fromthe stratified region, are proposed as possible explanationsfor copepod abundance increasing in advance of the summer peakin primary production.     

Predation, production and the organization of an estuarine copepod community

The copepod community of the estuaries near Beaufort, NC underwenta consistent seasonal succession from a spring assemblage dominatedby the medium-sized copepod Acartia tonsa (1 mm) to a summer—fallassemblage dominated by the small-bodied copepods Parvocalanuscrizsrirostris and Oithona colcarva (both 0.5 mm). However,in enclosure experiments during this period, A. tonsa dominatedthe community, due to higher growth rates and its predationon the nauplii of other species. Nutrient additions enhancedthe dominance by A. tonsa. The decline in abundance of A. tonsain the estuary was associated with increased abundance of planktivorousanchovies and silversides. In other enclosure experiments, planktivorousfish eliminated A. tonsa and other large copepods, althoughthey persisted in enclosures lacking fish. I conclude that predationby size-selective planktivorous fish prevents dominance by A.tonsa during summer—fall. ¹Present address: Institute of Marine Sciences, University ofNorth Carolina, 3407 Arendell Street, Morehead City, NC 28557,USA     

Mesozooplankton community indicates climate changes in a shelf area of the inner Bay of Biscay throughout 1988 to 1990

The mesozooplankton community was monitored at both coastaland offshore sites of the Basque shelf (inner Bay of Biscay)during 1988–1990 This was considered of interest becausethe local climate change toward dry conditions was found tobe responsible for changes in shelf-water properties. In additionto seasonal changes, interannual variations in mesozooplanktonabundance and composition were evident, and primarily relatedto the year-to-year increase in water temperature and salinity.The increasing trends in copepod dominance, coupled with theincreasing abundance of species with oceanic affinity (e.g.Euchaeta hebes), were presumably a response to the increasein oceanic features in the shelf area. In the same way, thesubstantial year-to-year increase in the abun dance of specieswith summer-autumn development was attributed to the progressivewarming. Among these species, the case of Temora stylifera isdiscussed specifically because this species was very scarcein 1988, but showed unusual high abundances for the study area,and dominated the summer-autumn assemblage in 1990. We thinkthat T.stylifera can be a key species in monitoring climate-oceanologicalchanges affecting the Bay of Biscay. Results are also discussedin the context of the long climate-oceanological series forthe Bay of Biscay, and its biogeographical status.     

The occurrence of Calanus finmarchicus (Gunnerus) and Calanus helgolandicus (Claus) in the western Irish Sea

The seasonal abundance of Calanus finmarchicus and Calanus helgolandicusin the North Channel and stratified region of the western IrishSea is reviewed using data collected between 1992 and 1996.Both species occur in the western Irish Sea, but were more abundantin the stratified region during spring. Increased abundanceduring May/June was attributed to an increase in copepoditestages. Calanus helgolandicus dominated in both regions, exhibitingspring and autumn peaks in abundance in the stratified region.It is argued that the presence of ripe females and naupliarstages in the stratified region is evidence of an in situ breedingpopulation, rather than advection of individuals from populationcentres outside the Irish Sea. The lack of geographical separationof the two species in the western Irish sea, and reports thatboth species occur in the Celtic Sea and Malin Shelf, limitthe use of either species as indicators of exchange processesbetween the Irish Sea and neighbouring waters.     

The production and ingestion of faecal pellets by nauplii of marine calanoid copepods

The downward transport of organic matter as zooplankton faecalmaterial is influenced by copepods which fragment, ingest andrecycle some of the pellet contents. Most of this activity hasbeen attributed to the later copepodite stages and the adults,but little is known about the role of nauplii. Stage-relateddefaecation rates during the naupliar development of two speciesof copepod, Calanus helgolandicus and Pseudocalanus elongatus,were quantified in a series of laboratory experiments. The productionof faecal material commenced soon after the appearance of theNIII in both species and increased throughout naupliar development.The causes of the increase were the formation of larger pelletsby later stages in Calanus and an increased rate of productionby Pseudocalanus. Calanus nauplii, when supplied with algalfood at concentrations that would support full naupliar development,ingested or broke up the pellets of the smaller Pseudocalanusspecies at rates of 1.15 pellets nauplius^–1 h^–1This consumption increased to 2.96 pellets nauplius^–1h^–1 when the concentration of algal food was reduced toa limiting level. Pseudocalanus was not able to consume thepellets of Calanus. Ingestion of Pseudocalanus faecal pelletsby Calanus could supply a nutritional benefit to a food-limitednauplius.     

Population biomass, feeding, respiration and growth rates, and carbon budget of the scyphomedusa Aurelia aurita in the Inland Sea of Japan

We investigated the seasonal occurrence, wet : dry : carbon: nitrogen weight ratios, population biomass, gastric pouchcontents, and rates of feeding, growth and respiration of thescyphomedusa Aurelia aurita in the central part of the InlandSea of Japan. Aurelia aurita medusae began to appear in January/Februaryas ephyrae, reached annual maximum body size in July/August,and disappeared, presumably due to death, by November. Initialslow growth in early spring was followed by a period of exponentialgrowth (mean growth rate: 0.069 d^–1) between April andJuly. In the Ondo Strait, which is characterized by strong tidalmixing, the A. aurita population (mean carbon biomass: 66.0mg C m^–3) overwhelmingly dominated the zooplankton-communitybiomass (mean biomass of micro- and mesozooplankton: 23.7 mgC m^–3) between May and early August The gastric contentanalysis revealed that A. aurita ate almost all micro- and mesozooplankters,of which small copepods were most important. On the basis ofdigestion time for small copepods (60 min) and their abundancein the gastric pouch of field-collected A. aurita, we determinedthe weight specific feeding rates and clearance rates. The formerincreases linearly with increasing copepod abundance, but thelatter was relatively constant irrespective of the food supply.We also measured the respiration rates of A. aurita and expressedthem as functions of body weight and temperature. These physio-ecologicalparameters enabled us to construct the carbon budget of theA. aurita population typical of early summer in the Ondo Strait.Predicted population-feeding rate (6.07 mg C m^–3 d^–1)was higher than the population-food requirement for both metabolismand growth (4.55 mg C m^–3 d^–1), indicating thatfood supply was sufficient to sustain the observed growth rate.This feeding rate was equivalent to 26% of micro- and mesozooplanktonbiomass, a significant impact on zooplankton.     

Zooplankton diversity and the predatory impact by larval and small juvenile fish at the Fisher Banks in the North Sea

The biomass and diversity of the mesozooplankton and fish larvaecommunity were investigated across a frontal zone in the centralNorth Sea in the early summer, to investigate whether larvalfish predation is a regulator of mesozooplankton production.Pronounced changes in the mesozooplankton community were observedacross the front off the Jutlandic coast. The biomass and thediversity of copepods changed across the front as the populationof Calanus finmarchicus became abundant in the deeper water.The crustaceans (Acartia spp. and Evadne spp.) and polychaetelarvae dominated the mesozooplankton in the coastal water. Thebiomass of fish larvae was dominated by gadoid larvae. As inthe copepods, a shift in fish diversity was observed in thefrontal zone. On the coastal side of the front, whiting (Merlangiusmerlangus) dominated the biomass, while offshore from the frontwhiting were absent and cod (Gadus morhua) was the dominantlarval fish species on the deeper stations. The present investigationdemonstrates two different trophic pathways related to hydrographyin the central North Sea. First, in the shallow coastal water,the abundant small neritic copepods are predominately predatedby whiting, while in the offshore region the larger oceaniccopepods are predated by cod larvae. However, the predationpressure by the fish larvae was in general low (<10%) relativeto copepod production per day. Consequently, in the early summer,the copepod production potentially results in a build-up ofcopepod biomass on both sides of the front.     

Seasonal Dynamics of Mesozooplankton in Brazilian Coastal Waters

The composition, numerical abundance and seasonal distribution of mesozooplankton, in addition to copepod biomass, were studied in the São Sebastião Channel (SSC) during different seasonal periods and hydrographic regimes. Two stations were sampled every 3 months from January 1996 to July 1997 and also in July 1998 and January 1999. Temporal differences in oceanographic conditions in the SSC were observed; there was clear seasonal variation in the thermohaline structure. Phytoplankton composition, standing-stock and biomass also showed consistent seasonal variation, peaking during summer due to advection of cold and nutrient-rich South Atlantic Central Water (SACW). Sixty-nine mesozooplankton taxa were identified and counted. Mesozooplankton density also increased during summer, ranging from 93 to 12,774 ind  m^?3. In contrast, a unimodal peak of copepod biomass was recorded during winter and under the influence of the oligotrophic Coastal Water (CW), suggesting that copepod biomass at SSC is driven by resource control. There was no significant evidence of seasonal pattern in overall community composition or evidence of changes due to water mass advection. The mesozooplankton community was continuously dominated by coastal and warm-water species, particularly the copepods Acartia lilljeborgi, Paracalanus aculeatus, P. quasimodo, Parvocalanus crassirostris, Corycaeus giesbrechti and Oithona oswaldocruzi, the cladocerans Penilia avirostris and Evadne tergestina, in addition to the appendicularians Oikopleura longicauda and O. dioica, all of which are perennial species in the SSC. This taxonomic composition may represent an efficient biotic structure for rapid recycling of primary production during periods of SACW influence.     

Fecundity limitation of Calanus helgolandicus, by the parasite Ellobiopsis sp.

The effect of the parasite Ellobiopsis sp., on the fecundityof Calanus helgolandicus and Calanoides carinatus in the Bayof Biscay, was investigated in May 2003. An average of 6.8%of C. helgolandicus females were infected with Ellobiopsis sp.,whereas none of the C. carinatus were found to be infected.An objective method of estimating gonad development was appliedto quantitatively measure the effect of the parasitism on thereproduction of the copepod. Parasitism by Ellobiopsis sp. hasthe potential to reduce the fecundity of C. helgolandicus females.     

Existence and significance of Acartia grani resting eggs (Copepoda: Calanoida) in sediments of a coastal station in the Alboran Sea (SE Spain)

The seasonal abundance of copepod eggs in bottom sediments ofMalaga harbour was documented during an annual cycle betweenJune 1989 and June 1990. The concentration noted ranged from6.6.10⁶ to 0.1910⁶ m^–2. The number of adults in thewater column and eggs recovered from the bottom sediments fluctuatedmarkedly with season. The highest egg abundance in sedimentswas found coincidentally with the early population growth season,although the values remained high until the adults disappearedfrom the water column. During the winter, the number of eggsdecreased until the development of the next copepod population.When bottom sediments were incubated in the laboratory, a largenumber of Acartia nauplii hatched, which showed that eggs laidin the autumn remain viable at the bottom until late spring,thus providing a large pool of potential recruits for the planktonicpopulation. ¹Present address: Departamento de Biologa Animal, Vegetal yEcologa, Facultad de Ciencias Experimentales, Universidad deMan Paraje Las Lagunillas s/n, 23071 Man, Spain     

Response of ciliates and Cryptomonas to the spring cohort of a cyclopoid copepod in a shallow hypereutrophic lake

The impact of a cyclopoid copepod population on the protozoacommunity (two ciliate categories and Cryptomonas) was assessedweekly during the spring cohort of Cyclops vicinus (one monthduration) in hypereutrophic Lake Søbygård by insitu gradient experiments with manipulation of ambient zooplanktonabundance. As C.vicinus always made up >92% of the zooplanktonbiomass, the response of protozoa is assumed to be a resultof predation by the copepod. Significant effects of copepodbiomass on protozoa net population growth rates were obtainedin the four experiments. Copepod clearance rates were significantlyhigher on oligotrichs than on prostomatids and Cryptomonas butdeclined for all three protozoa categories during the firstthree weeks of the copepod cohort, probably because of the changein developmental instar composition of the copepod population.Grazing impact on protozoa at ambient copepod abundance wasconsiderable (range, 0.05–0.87 day^–1) and could,together with the estimated reproductive potential of protozoans(range, –0.20–0.87 day^–1), account for thedecline in abundance and biomass of protozoa during the cohortdevelopment. Carbon flow from the protozoa to C.vicinus (range,2.8–23.5 µg C l^–1 day^–1) documents thepresence of a trophic link between protozoa and the spring cohortof C.vicinus in Lake Søbygård.     

Seasonal variation in the species composition of tintinnid cilates in Hiroshima Bay, the Seto Inland Sea of Japan

Seasonal changes in the species composition of tintinnid ciliateswere examined based on time-series samples taken at 2 week intervalsover a 3 year period in Hiroshima Bay, the Seto Inland Sea ofJapan. The maximum abundance of total tintinnids over the entireperiod was 5.7 x 10³ indi viduals l^– Among 32 speciesidentified, a consistent seasonal occurrence was recognizedin 22 species. The relationships between various environmentalfactors and the abundance of each species of tintinnids wereanalyzed using principal component analysis From this analysis,the abundance of many tintinnids was revealed to be associatedwith temperature, the <20 µm size fraction of chlorophylla and water column stability, but not with the <20 µmsize fraction of chlorophyll a, nor with salinity. From theseresults, tintinnid species were divided into five associationtypes: species whose abundance increased with increasing temperature,decreasing temperature, nanophytoplankton abundance, increasingwater column mixing, or increasing water stratification coupledwith low temperature.     

Production of Penilia avirostris in Kingston Harbour, Jamaica

The cladoceran Penilia avirostris is one of the more abundantand widespread members of the crustacean zooplankton in nearshoretropical and subtropical waters. Its abundance, biomass, fecundity,development rate and production were estimated in Kingston Harbour,Jamaica, during an 18 month period. Mean annual abundance ofPenilia was 1821 m^–3, while biomass (excluding eggs/embryos)was 2.87 mg ash-free dry-weight (AFDW) m^–3 (43.1 mg AFDWm^–2), accounting for 13% of the copepod community biomass.Fecundity increased with body size. There was no clear seasonalpattern of abundance, size or fecundity, nor were physical orbiological variables correlated to these variations. Developmenttime averaged 20.5 h for juveniles and 41.4 h for adult femalesduring incubations; there was no clear evidence of a diel patternto molting. Growth rate appeared to be exponential, with correspondingsomatic growth rates, averaging 0.27 day^–1 for juveniles,and 0.34 day^–1 for somatic plus reproductive growth inadult females. Annual production was estimated as 173 kJ m^–2year^–1,     

Direct Somatic Embryoid Formation on Immature Embryos of Trifolium repens, T. pratense and Medicago sativa, and Rapid Clonal Propagation of T. repens

By direct somatic embryogenesis in vitro a clone of asepticplantlets can be raised from a single immature embryo of Trifoliumrepens (white clover) within about 6 weeks of pollination. Embryoidsare induced directly from intact zygotic embryonic tissue ona culture medium containing 0·025 or 0·05 mg 1^–1BAP and 1·0 g 1^–1 yeast extract. Similar directsomatic embryogenesis has also been achieved for Trifolium pratense(red clover) and Medicago sativa (lucerne). Applications ofembryo propagation by direct somatic embryogenesis are discussed,particularly in relation to multiple screening of host genotypesfor analysis of host/pathogen and legume/Rhizobium interactions. Trifolium repens L., Trifolium pratense L., Medicago sativa L., clover, lucerne, tissue culture, embryoid, somatic embryogenesis, legumes     

There is No Somatic Meiosis in Embryogenic Leaves of Cichorium

Several papers dealing with carrot cell cultures describe meiosis-likedivisions and haploid cells prior to somatic embryogenesis.We have studied the first division in embryogenic mesophyllcells of a diploidCichorium intybus L. and of a tetraploid hybridC.intybus L.xC. endivia L. which undergo direct somatic embryogenesisfrom single cells when leaf fragments are placed in a liquidagitated inductive medium (modified MS with 1x10^-7M NAA and2.5x10^-6M 2-iP), in darkness, at 35°C. MicrosporogenesisinC. intybus provided aspects of meiosis for comparison. Inleaves incubated in inductive conditions, DAPI staining of nucleishowed normal mitosis on days 3–6; about 0.6% cells inprophase had undergone spontaneous endoreduplication leadingto a tetraploid somatic embryo. Immunocytochemistry of tubulinrevealed the constant presence of a preprophase band, as ina normal mitosis. The first pluricellular somatic embryos becamevisible on day 5 of culture. Flow cytometric determination ofnuclear DNA on days 4, 5 and 6 did not show any peak correspondingto the 1C DNA level for the diploid plant or to the 2C DNA levelfor the tetraploid. Instead there was a weak but constant peakat the 4C and 8C levels. We conclude that inCichorium leaves,the first division of somatic embryogenesis is a normal mitosis,with a small shift to endoreduplication. In our opinion, somaticmeiosis is not a prerequisite during direct somatic embryogenesis. Cichorium ; chicory; somatic embryogenesis; cell division; flow cytometry; tubulin     

Direct Somatic Embryogenesis in Roots of Cichorium: Is Callose an Early Marker?

Direct somatic embryogenesis can be obtained from epidermaland cortical cells in roots from in vitro Cichorium plantlets.The first embryogenic cells are seen after six days of culturein darkness, at 35 °C, in a liquid medium supplemented withNAA (1 x 10^–7 M), 6-dimethylallyl-amino-purine (2·5x 10^–6 M), sucrose (0.03 M) and glutamine (1·7x 10^–3 M). Embryogenic cells undergo first a linear andthen a globular segmentation, with increasing cytoplasmic density.These cells and young embryoids show aniline blue fluorescence.SEM allows the same microglobular pattern to be seen on thesurface of young embryoids and on young microspores of Cichoriumused as controls. In this root system, callose deposition seemsto be an early marker in somatic embryogenesis. Somatic embryogenesis, callose, Cichorium     

Auxin and heat shock activation of a novel member of the calmodulin like domain protein kinase gene family in cultured alfalfa cells

A calmodulin like domain protein kinase (CPK) homologue wasidentified in alfalfa and termed MsCPK3. The full-length sequenceof cDNA encoded a 535 amino acid polypeptide with a molecularweight of 60.2 kDa. The deduced amino acid sequence showed allthe conserved motifs that define other members of this kinasefamily, such as serine-threonine kinase domain, a junction regionand four potential Ca²⁺-binding EF sites. The recombinant MsCPK3protein purified from E. coli was activated by Ca²⁺and inhibitedby calmodulin antagonist (W-7) in in vitro phosphorylation assays.The expression of MsCPK3 gene increased in the early phase ofthe 2,4-D induced alfalfa somatic embryogenesis. Heat shockalso activated this gene while kinetin, ABA and NaCl treatmentdid not result in MsCPK3 mRNA accumulation. The data presentedsuggest that the new alfalfa CPK differs in stress responsesfrom the previously described homologues and in its potentialinvolvement in hormone and stress-activated reprogramming ofdevelopmental pathways during somatic embryogenesis. Key words: Medicago sativa, CPK, stress, 2,4-D, phosphorylation, somatic embryogenesis.