Calcium exchange and the measurement of calcification rates in the calcareous coralline red alga Amphiroa foliacea

⁴⁵Ca washout data of living thalli of Amphiroa foliacea Lamouroux in the light and dark show that there are three kinetically distinct Ca²⁺-exchanging compartments with approximate half-times (t 1/2) of 300, 20 and 3 min. The two slower compartments appear to be exchange from organic Ca²⁺-binding components of the cell wall, while the fast compartment probably represents exchange on the CaCO₃ crystal surface. Killed and decalcified thalli have a fourth compartment, with a t 1/2 of 20 to 35 min (other compartment half-times are 300, 40, 3 min), which has been identified as the greatly increased intercellular space produced during drying and decalcification. The ⁴⁵Ca and ¹⁴C uptake data show that a large proportion of the label initially taken up is into compartments other than the CaCO₃. As a result of this uptake, binding, and exchange of radioisotope, significant errors occur during the measurement of calcification rates, unless a kinetic analysis is carried out. Using such a technique, CaCO₃ calcification rates of A. folicea were measured with ⁴⁵Ca or ¹⁴C as tracers. Light stimulates calcification by up to 2.6 times, depending upon the age of the plant. Young segments have a markedly higher rate of calcification and photosythesis than do the older segments.     

A method for the determination of copepod feeding rates during short time intervals

An experimental technique for measuring the feeding behavior of copepods on small time scales is important because grazer/micro-patch interactions in the ocean are probably short (minutes to hours). By placing copepods with empty guts into a known food concentration and monitoring the rate of increase in gut contents until defecation begins, usually 20 to 100 min, the volume of water swept clear of particles per min can be calculated. This process is easily quantified by using phytoplankton as food and measuring the gut contents by fluorometry. The utility of this technique is demonstrated in the laboratory and at sea.     

Determination of photosynthetic rates for the marine algae Fucus vesiculosus and Laminaria digitata

A continuously recording, flow-through oxygen electrode system for the measurement of oxygen exchange is described and applied to an investigation of photosynthetic rates in the marine algae Fucus vesiculosus L. and Laminaria digitata (Huds.) Lam. The photosynthetic rate (mg O₂.g dry weight^-1.h^-1) at 15°C and 21.5 mW.cm^-2 (usually just saturating) ranges in F. vesiculosus from 1.20 in basal portions of the thallus to 9.27 at the apices and in L. digitata from 1.19 mg O₂ at the thallus base to 3.97 mg O₂ at distances of several centimetres behind the upper thallus margin. This variation is reduced when the photosynthetic rate is expressed in terms of fresh weight or surface area.This research was carried out while one of us (R.J.K.) was an Alexander von Humboldt fellow at the University of Kiel, and is part of the programme Sonderforschungsbereich 95, Wechselwirkung Meer-Meeresboden, Universität Kiel.     

Parthenogenetic reproduction of Diaphanosoma celebensis (Crustacea: Cladocera). Effect of algae and algal density on survival, growth, life span and neonate production

The cladoceran Diaphanosoma celebensis Stingelin is reported on for the first time from Indian waters (Mandovi estuary, Goa). Amictic females were maintained in the laboratory (temperature 24 ± 1 °C and salinity 17 psu) for three successive generations in order to follow the parthenogenetic reproductive behaviour, growth, survival and neonate production. The mean life span and body length of adult females in the three generations showed some variations and ranged from 9 to 12.5 d and 842 to 932 μm, respectively. The mean length of the neonates produced also varied (283 to 446 μm) in the three generations. Cladoceran preference for three phytoplankton food sources, i.e. Isochrysis galbana (Parke), Chaetoceros calcitrans (Paulsen) and Tetraselmis gracilis (Kylin), was determined. Growth was faster in the initial stage with all three diets but slowed down in later life. Increased food concentrations resulted in higher neonate production but reduced the life span of females. However, long-term feeding experiments revealed that the percentage survival was high with I. galbana and low with C. calcitrans. Received: 23 June 1999 / Accepted: 20 September 1999     

Impact of calcium and magnesium on growth and morphological acclimations of Nitella: implications for calcification and nutrient dynamics

The impact of calcium (Ca) and magnesium (Mg) on the growth and morphology of a Charophyte, Nitella pseudoflabellata, and the influence of Mg on calcification and phosphorous (P) speciation were studied in laboratory experiments for variable concentrations (≤q120 mg · L^?1) of Ca and Mg. It was clearly identified that Mg aided shoot elongation. An increase in Ca concentrations produced intensified shoot elongation also, but at a lesser rate than the equivalent levels of Mg. Depending on the availability of Ca and Mg, the morphological appearance differed significantly, suggesting significant levels of ecoplasticity. Furthermore, Mg was observed to produce less calcite encrustation. Plant P-speciation suggested a higher Mg concentration corresponding to a more water-soluble and less carbonate-bound P fraction. This indicates that upon senescence and decomposition, a large fraction of P is supplied to the water column; ultimately behaving similar to a typical vascular plant.     

Application of a 15N tracer method to the study of dissolved organic nitrogen uptake during spring and summer in Chesapeake Bay

The dissolved organic nitrogen (DON) pool in marine waters contains a diverse mixture of compounds. It is therefore difficult to accurately estimate planktonic uptake of DON using the limited number of radiolabeled compounds commercially available. We describe a method to estimate DON uptake rates using ¹⁵N-labeled DON recently released from phytoplankton. To make ¹⁵N-labeled DON, we incubated surface water with ¹⁵NH ₄ ⁺ and then isolated the DON, including any recently released DO¹⁵N, with ion retardation resin. This DON was then added to a freshly collected water sample from the same environment to quantify the rate of DON uptake. The technique was applied to investigate rates of DON uptake relative to inorganic nitrogen in the mesohaline Chesapeake Bay during May 1990 and August 1991. The May experiment took place after the spring bloom, and rates of DON uptake [ranging from 0.31 to 0.53 g-atom (g-at) Nl^-1 h^-1] often exceeded rates of NH ₄ ⁺ and NO ₃ ^- uptake combined. The rates of DON uptake at this time were higher than estimated bacterial productivity and were not correlated with bacterial abundance or bacterial productivity. They were, however, correlated with rates of NO ₃ ^- uptake. In May, we estimate that only 7 to 32% of DON uptake was a result of urea utilization. In contrast, in August, when regenerated nutrients predominate in Chesapeake Bay, rates of DON uptake (ranging from 0.14 to 0.51 g-atom Nl^-1 h^-1) were an average of 50% of the observed rates of NH ₄ ⁺ uptake. Consistent with the May experiment, rates of DON uptake were not correlated with bacterial production. A sizable fraction of DON uptake, however, appeared to be due to urea utilization; rates of urea uptake, measured independently, were equivalent to an average of 74% of the measured rates of DON uptake. These findings suggest that, during both periods of study, at least a fraction of the measured DON uptake may have been due to utilization by phytoplankton.     

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.     

Simplified method for the determination of acid-soluble sulfides in marine sediments

A simple, rapid method for the measurement of acid-soluble sulfides from marine sediments, in the range of 0 to 350 g sulfide, was developed. Using the mean of 4 replicates, the maximum sensitivity of the method is ±5.48 g. The method is based on iodometric back titration after the reaction of a standard iodine solution with the sulfide, which is precipitated on paper wicks as zinc, sulfide. The mean coefficient of variation, based on raw titration data, is 0.71% for aliquots from field samples, 0.41% for standards, and 0.21% for blanks. The method is used to compare zinc acetate-fixed subsamples, one group having been stored for 1 month in a refrigerator at 5°C and the other having been analyzed immediately after collection. No significant differences in sulfide content between subsamples, at the 95% confidence level, were noted.     

Effect of temperature and light on the growth of micro- nano- and pico-plankton: impact on algal succession

The influence of light and temperature on the phytoplankton succession in a temperate sea area was investigated in laboratory experiments with natural assemblages of micro-, nano-, and picoplankton collected from the northern Baltic Sea during 1989 and 1990. Respiration increased from 0 to 30°C in all groups of phytoplankton, while gross photosynthesis stabilised at 10 to 15°C. Light saturation occurred at 25 to 75 mol quanta m^-2 s^-1, indicating low light adaptation of the algae. Picoplankton showed the strongest temperature response, and at temperatures above 10°C picoplankton obtained a higher biomass specific photosynthesis than that of the other groups. Different light treatments had no effect on the species composition in experiments with natural algal assemblages, while different temperature conditions had a marked effect. With a temperature increase from 0 to 10°C, the algal community changed from a typical spring community, with diatoms and dinoflagellates, to a summer community, dominated by mixotrophic nanoflagellates. The small, or often non-existing, autumn bloom in the sea area studied, can be explained by short day lengths combined with relatively high temperatures, causing high community respiration rates and low gross photosynthesis, resulting in a negative energy balance. The net energy gain depends on a differential temperature effect on gross photosynthesis and endogenous respiration in various plankton groups. This gives the phytoplankton groups diverse competitive advantages during different seasons and thus may be an important factor in controlling algal succession.     

A method for the assessment of light-induced oxidative stress in embryos of fucoid algae via confocal laserscan microscopy

A method was developed for measurement of active oxygen production in embryonic stages of the brown seaweed Fucus spiralis, using the label CM-DCFH-DA. Active oxygen species convert the label into the green fluorescent CM-DCF (exc/em 488/530 nm) that is detected via confocal laserscan microscopy and quantitative image analysis. Loading of the label did not harm the embryos; loading efficiency was age-independent, and the esterases needed for conversion to CM-DCFH were not inhibited by the effective UV dose (2 W m⁻²) applied in the experiments. After correction for daily variation of the laser power, and calibration with DCF standard solutions, this automated analysis of confocal images rendered active oxygen concentrations in fucoid embryos (μM DCF). An experiment was designed for the assessment of active oxygen production following irradiance stress in the light-sensitive embryos. Dim-light-acclimated, 1-, 2- and 4-day-old embryos were transferred for 60 min to defined high-light conditions (4π-irradiance 300 μmol photons m⁻² s⁻¹), optionally without UV radiation, including UVA, or including UVA plus UVB. PSII yield measurements (PAM fluorometer) were carried out in order to assess the degree of photoinhibition under these light conditions. The imposed light stress initially caused a rapid decrease of the PSII yields (Φ_P). With increasing embryo age, minimum Φ_P values attained under light stress remained higher. Consequently, electron transport rates (ETR) would increase with embryo age, i.e., with the development of their photosynthetic apparatus. Active oxygen production increased with ETR, and when UVB was included, relatively greater amounts of active oxygen were produced. A slow, second-phase decrease of Φ_P under light stress that was proportional to active oxygen production indicated that some photooxidative damage was caused, in particular during UVB exposure. Recovery from light stress was a rapid process in the absence of UVB; in such cases Φ_P was almost restored to the initial values within 60 min. The relative state of recovery of Φ_P was correlated with both the effective UV dose and active oxygen production rate (DCF). Recovery was slowest in embryos exposed for 60 min to an experimental UVB dose, which was representative of a situation at low tide, on a sunny day. The results suggest that active oxygen may cause an in situ inhibition of growth of the earliest life stages of F. spiralis. Received: 26 January 2000 / Accepted: 4 September 2000     

A sensitive spectrophotometric method for the determination of arsenic in environmental samples

We developed a cost-effective and sensitive spectrophotometric method for the determination of arsenic at trace level using a new reagent, leuco malachite green. Here we show that, arsenic reacts with potassium iodate in acidic conditions to liberate iodine, and the liberated iodine selectively oxidizes leuco malachite green to malachite green dye. We studied the Beer’s law at 617 nm, which showed linearity over the concentration range 0.09–0.9 μg ml⁻¹ of arsenic. We show that the molar absorptivity, Sandell’s sensitivity and detection limit of the method are 6.1 × 10⁴ l mol⁻¹ cm⁻¹, 0.0012 μg cm⁻² and 0.025 μg ml⁻¹, respectively. We applied the developed method for the determination of arsenic in environmental samples.     

Effect of temperature and temperature adaptation on calcification rate in the hermatypic coral Pocillopora damicornis

Using ⁴⁵Ca incorporation into the coral skeleton as a measure of calcification rate, the effect of temperature on clacification rate was studied in the hermatypic coral Pocillopora damicornis. Both immediate and long-term (adaptation) effects were investigated. Temperature has a marked effect on rate — an effect that varies depending on the temperature history of the coral (i.e., temperature adaptation occurs). P. damicornis showed both 27° and 31°C temperature optima, one or the other being dominant depending on the natural water temperature to which the coral was adapted. The two optimum temperatures may indicate two isoenzymes or two alternate metabolic pathways involved in the calcification process.     

Caste determination and differential diapause within the first brood of Halictus rubicundus in New York (Hymenoptera: Halictidae)

Summary The partially bivoltine, primitively eusocial sweat bee Halictus rubicundus produces two female castes, gynes and non-gynes, in its first brood in New York. Castes in this brood differentiate within the first few days of adult life, with gynes leaving the population to overwinter as early as mid-June (non-gynes further differentiate into replacement queen and worker subcastes, with older females typically dominant). Analysis of possible mechanisms of caste determination reveals that although gynes average significantly larger than non-gynes within the first brood, this appears largely due to a late mean emergence data coupled with an increase in the size of emerging females over the course of brood emergence, rather than a causal relationship. A strong correspondence between male abundance (relative to newly-emerged females) and the pattern of gyne production, along with data from dissections, suggests that females that mate when young become diapausing gynes, while those that do not mate promptly become non-gynes and do not diapause even though many mate later. Although alternatives to this simple mechanism cannot be ruled out entirely, it nevertheless offers profound implications for theoretical and empirical understanding of the evolutionary origins of the worker caste.     

Light regime in an Arctic fjord: a study related to stratospheric ozone depletion as a basis for determination of UV effects on algal growth

Solar radiation as a primary abiotic factor affecting productivity of seaweeds was monitored in the Arctic Kongsfjord on Spitsbergen from 1996 to 1998. The radiation was measured in air and underwater, with special emphasis on the UV-B (ultraviolet B, 280–320 nm) radiation, which may increase under conditions of stratospheric ozone depletion. The recorded irradiances were related to ozone concentrations measured concurrently in the atmosphere above the Kongsfjord with a balloon-carried ozone probe and by TOMS satellite. For comparison, an ozone index (a spectroradiometrically determined irradiance of a wavelength dependent on ozone concentration, standardized to a non-affected wavelength) was used to indicate the total ozone concentration present in the atmosphere. Weather conditions and, hence, solar irradiance measured at ground level were seldom stable throughout the study. UV-B irradiation was clearly dependent on the actual ozone concentration in the atmosphere with a maximal fluence rate of downward irradiance of 0.27 W m⁻² on the ground and a maximal daily fluence (radiation exposure) of 23.3 kJ m⁻². To characterize the water body, the light transmittance, temperature and salinity were monitored at two different locations: (1) at a sheltered shallow-water bay and (2) at a wave-exposed, deep-water location within the Kongsfjord. During the clearest water conditions in spring, the vertical attenuation coefficient (K _d) for photosynthetically active radiation (PAR) was 0.12 m⁻¹ and for UV-B 0.34 m⁻¹. In spring, coinciding with low temperatures and clear water conditions, the harmful UV radiation penetrated deeply into the water column and the threshold irradiance negatively affecting primary plant productivity was still found at about 5–6 m depth. The water body in spring was characterized as a Jerlov coastal water type 1. With increasing temperature in summer, snow layers and glacier ice melted, resulting in a high discharge of turbid fresh water into the fjord. This caused a stratification in the optical features, the salinity and temperature of the water body. During melt-water input, a turbid freshwater layer was formed above the more dense sea water. Under these conditions, light attenuation was stronger than defined for a Jerlov coastal water type 9. Solar radiation was strongly attenuated in the first few metres of the water column. Consequently, organisms in deeper water are protected against harmful UV-B radiation. In the surface water, turbidity decreased when rising tide caused an advection of clearer oceanic water. In the course of the summer season, salinity continuously decreased and water temperature increased particularly in shallow water regions. The impact of global climate change on the radiation conditions under water and its effects on primary production of seaweeds are discussed, since organisms in the eulittoral and upper sublittoral zones are affected by UV radiation throughout the polar day. In clearer water conditions during spring, this may also apply to organisms inhabiting greater depths. Received: 20 June 2000 / Accepted: 17 October 2000     

A fast and reliable method for quantitative determination of total mercury in vegetables

A cloud-point extraction (CPE) process using the nonionic surfactant, polyethylene glycol tert octylphenyl ether (Triton X-114) was employed for determination of Hg(II) ions in aqueous solutions. The method is based on the ion-pairing reaction of Hg(II) with Pyronin B (PyrB⁺) in the presence of excess iodide at pH 6.0 and extraction of the complex formed. The chemical variables affecting CPE efficiency were studied, and the analytical characteristics of the method were obtained. The calibration curves were linear in the range of 1–40 μg L^?1 with the detection limits of 0.35 and 0.30 μg L^?1 at 556 and 521 nm. Selectivity was also tested. The coefficients of variation of the method are 2.4% and 5.2% for five replicate measurements of mercury at levels of 10 and 25 μg L^?1, respectively. The results obtained for two certified reference samples were in a good agreement with the certified values. The method was applied to the determination of total mercury in vegetable samples.     

Automated method for the determination of total arsenic and selenium in natural and drinking water by HG-AAS

A multicommutated flow system was designed and evaluated for the determination of total arsenic and selenium by Hydride Generation Atomic Absorption Spectrometry (HG-AAS). It was applied to the determination of arsenic and selenium in samples of natural and drinking water. Detection limits were 0.46 and 0.08 μg l⁻¹ for arsenic and selenium, respectively; sampling frequency was 120 samples h⁻¹ for arsenic and 160 samples h⁻¹ for selenium. Linear ranges found were 1.54–10 μg l⁻¹ (R = 0.999) for arsenic and 0.27–27 μg l⁻¹ (R = 0.999) for selenium. Accuracy was evaluated by spiking various water samples and using a reference material. Recoveries were in the range 95–116%. Analytical precision (s _r (%), n = 10) was 6% for both elements. Compared with the Standard Methods, APHA, 3114B manual method, the system consumes at least 10 times less sample per determination, and the quantities of acid and reducing agent used are significantly lower with a reduction in the generation of pollutants and waste. As an additional advantage, the system is very fast, efficient and environmentally friendly for monitoring total arsenic and selenium levels in waters.     

A new derivatization method for a determination of MX and related hydroxyfuranones in water

An extremely potent mutagen, 3‐chloro‐4(dichloromethyl)‐5‐hydroxy‐2(5/f)‐furanone (MX) is commonly present in chlorinated drinking water. Due to its high mutagenic activity and according to WHO guidelines its concentration should be controlled in drinking waters. Determination of MX is difficult due to ppt levels at which the compound usually exists in drinking waters. Derivatization with 2‐propanol is presented as a method which significantly lowers GC/MS detection level of MX. Suitability of 2‐propylation for derivatization of other hydroxyfuranones is also shown.     

Effect of nutrient enrichment in the field on the biomass, growth and calcification of the giant clam Tridacna maxima

Nutrients were added separately and combined to an initial concentration of 10 μM (ammonium) and/or 2 μM (phosphate) in a series of experiments carried out with the giant clam Tridacna maxima at 12 microatolls in One Tree Island lagoon, Great Barrier Reef, Australia (ENCORE Project). These nutrient concentrations remained for 2 to 3 h before returning to natural levels. The additions were made every low tide (twice per day) over 13 and 12 mo periods for the first and second phase of the experiment, respectively. The nutrients did not change the wet tissue weight of the clams, host C:N ratio, protein content of the mantle, calcification rates or growth rates. However, ammonium (N) enrichment alone significantly increased the total population density of the algal symbiont (Symbiodinium sp.: C = 3.6 · 10⁸ cell clam⁻¹, N = 6.6 · 10⁸ cell clam⁻¹, P = 5.7 · 10⁸ cell clam⁻¹, N + P = 5.7 · 10⁸ cell clam⁻¹; and C = 4.1 · 10⁸ cell clam⁻¹, N = 5.1 · 10⁸ cell clam⁻¹, P = 4.7 · 10⁸ cell clam⁻¹, N + P = 4.5 · 10⁸ cell clam⁻¹, at the end of the first and second phases of the experiment, respectively), although no differences in the mitotic index of these populations were detected. The total chlorophyll a (chl a) content per clam but not chlorophyll a per cell also increased with ammonium addition (C = 7.0 mg chl a clam⁻¹, N = 13.1 mg chl a clam⁻¹, P = 12.9 mg chl a clam⁻¹, N + P = 11.8 mg chl a clam⁻¹; and C = 8.8 mg chl a clam⁻¹, N = 12.8 mg chl a clam⁻¹; P = 11.2 mg chl a clam⁻¹, N + P = 11.3 mg chl a clam⁻¹, at the end of the first and second phases of the experiment, respectively). The response of clams to nutrient enrichment was quantitatively small, but indicated that small changes in inorganic nutrient levels affect the clam–zooxanthellae association. Received: 2 June 1997 / Accepted: 9 June 1997     

Application of an inclusion complex for determination of dithianon residues in water and fruits

A 3,3′-benzidine/β-cyclodextrin fluorescence derivative (BCDFD) was prepared and characterized by means of elemental analysis and ¹H NMR. An inclusion complex was formed between BCDFD and the pesticide dithianon based on the internal cavity of β-cyclodextrin (β-CD), and the binding constant (K_S ) of the inclusion complex was investigated. The inclusion complex could be applied for the determination of dithianon residues. Based on this, a sensitive method for the determination of dithianon was established. The linear dynamic range was from 0.63 to 12.5?mg?L^?1 with a correlation coefficient of 0.9927. The limit of detection for an aqueous solution of standard and the relative standard deviation (RSD) were 10.6?µg?L^?1 and 0.76%, respectively. The proposed method has been successfully applied for the determination of dithianon residues in water and fruit samples with satisfactory results, and recoveries in the range of 96.0–108% were obtained. The method is rapid, direct, economical, and sensitive for dithianon analysis.     

不同蒸腾作用对番茄幼苗吸收Pb、Cd的影响

蔬菜中重金属污染是人们关注的热点问题之一,其中Pb、Cd的污染在我国种植蔬菜的土壤中较为突出,蔬菜对Pb、Cd的吸收除了受到土壤中Pb、Cd含量的影响之外,蔬菜本身的蒸腾作用是否会对吸收过程产生作用.通过在人工气候箱中,调节环境的温度和湿度,形成不同的环境蒸汽压,使植物产生不同的蒸腾作用,利用水培实验研究了在不同蒸腾作用下两个番茄品种(Lycopersicon esculentu,品种名分别为FA-189和合作903)的幼苗对重金属Pb、Cd的吸收富集规律.结果表明,高蒸腾作用下植株Cd和Pb含量比低蒸腾作用下分别增加了1.47～1.73倍和1.25～1.75倍,单株的积累量则分别增加了1.71～3.18倍和1.67～2.21倍.通过分析蒸腾水量对吸收Pb、Cd的影响,表明番茄幼苗通过蒸腾作用拉力吸收进入体内的重金属的量占总吸收量的比例极小,一般在0.000 8%～0.2%之间,说明番茄幼苗对Pb和Cd的吸收主要为代谢吸收.同时在比较了番茄对Pb和Cd的被动吸收比例时,发现Pb较Cd的比例要高几倍到几十倍,说明番茄对Pb和Cd的吸收富集规律可能存在不同的机理.