Determination of 14C labelled carbonate in solution

A direct and a diffusion method to be applied when measuring ¹⁴C labelled carbon dioxide by productivity measurements in natural water have been described. The labelled carbonate was trapped by 1 N solutions of NaOH, KOH or Ba(OH)₂. Results comparable to those obtainable the biological method are achieved when using 100 l of a 1 N KOH solution as a fixation reagent. The radiocarbonate fixed in this way was found to be stable for several days when dried, cooled, and stored in a desiccator. The reliability of these methods was demonstrated by analysing solutions containing various amounts of ¹⁴C labelled carbonate.     

Note on the variability of heterotrophic activity measurements by the 14C method in sea water

Twenty five measurements in triplicate of heterotrophic activity in sea water showed good reproducibility. The mean coefficient of variation was 6.2%, which is lower than the coefficient of variation obtained with a similar method for primary production measurements. This result indirectly confirms that most heterotrophic activity can be attributed to a great number of small cells, i.e., bacteria which, if not free-swimming, are fixed to particles of smaller size than the average phytoplankton. The good reproducibility obtained also suggests that there was no extraneous contamination despite non-sterile working conditions.     

14C and 125I Labelling of Humic Material for Use in Environmental Studies

Humic and fulvic acids are present in nearly all natural waters. These acids are known to affect the transport of environmental contaminants such as metals and hydrophobic organics through the terrestrial environment. an understanding of their role in the transport of contaminants is therefore essential and is facilitated if the acid is labelled with a suitable radioactive label. This paper reports the use of ¹⁴C-methylamine and ¹²⁵I to label humic acid with either ¹⁴C or ¹²⁵I and investigates factors which affect the yield of these reactions. the stability and mobility of the labelled humic material through sand is also reported.     

Coral illumination through an optic glass-fiber: incorporation of 14C photosynthates

The pathways of ¹⁴C incorporation into the three major compartments of the coral body were analysed in colonies of Stylophora pistillata. We used the optic glass-fiber method to carry out two sets of experiments: in the first, 11 different colonies were sampled immediately after incubation; in the second, 3 colonies were returned to the reef at the termination of incubation for a further period of 29 h. Within the tissue compartment, significantly more ¹⁴C labeled products were incorporated into illuminated tips or bases than into unilluminated sections. Tips located above illuminated bases contained amounts of ¹⁴C products similar to unilluminated tips. Within the organic matrix compartment, illumination of tip or base segments again resulted in increased amounts of ¹⁴C fixation, and again unilluminated tips located above the illuminated bases did not accumulate more ¹⁴C photosynthates than other tips on the same branches. The absence of detectable translocation was also confirmed after a post-incubation period of 29 h, and raises questions as to the validity of the widely accepted theory of upward translocation. Within the skeletal carbonate compartment, although the results were associated with a high coefficient of variation, significantly more ¹⁴C accumulated in the tips than in the bases. Illumination of tips or bases did not enhance ¹⁴C uptake. A light-independent carbon assimilation (dark fixation) is significant in S. pistillata within the three tested compartments (the tissue, the organic matrix of the skeleton, and the skeletal carbonate). It is suggested that the dark fixation process in corals in a result of accumulation of respiratory CO₂ and CO₂ from sea water as malate or other titratable acids during the night. During the day these acids are broken down, releasing free CO₂ for C₃ pathway photosynthesis.     

Resource translocation in a marine bryozoan: quantification and visualization of 14C and 35S

Colonial marine invertebrates are characterized by their ability to share resources among the modules of a colony. In most colonial groups, but particularly the Bryozoa, the dynamics of resource transport among modules is unknown. We developed radioisotope techniques to visualize and quantify the movement of carbon and sulfur-based compounds within colonies of the marine bryozoan Membranipora membranacea. The research was conducted in 1991 and 1992 in Friday Harbor, Washington, USA. Autoradiography, using X-ray film, was used to visualize the transport of both ¹⁴C and ³⁵S, and a liquid scintillation counter was used to quantify transport of metabolites. We were able to localize feeding by introducing 10 l aliquots of labelled algal cells with a microinjection syringe into a containment ring on the surface of the colony. The labelled cells were consumed by zooids feeding within the ring, but not by those outside. In time-course within the ring, but not by those outside. In time-course experiments, 15% of the ingested carbon radioisotope was transported from a source in the center of the colonies to the growing edges over a period of 48 h. Approximately 10% of the sulfur was transported from central to edge regions of colonies over 72 h. Transport of carbon isotope was unidirectional in all experiments, irrespective of whether colonies were fed near the edge or the center. Pulse-chase experiments revealed that up to 46% of the initial ¹⁴C radioisotope was lost from the colony to respiration and egestion in the 24 h following ingestion.     

14C of grasses as an indicator of fossil fuel CO2 pollution

Measuring the amount of fossil fuel carbon stored in the vegetation is now crucial to understand the mechanisms ruling climate changes. In this respect, highly polluted areas such as major towns represent “natural” laboratories because fossil fuel CO₂ (¹⁴C-free) is isotopically distinct from mean atmospheric CO₂ (¹⁴C-labeled). Here, a¹⁴C study of urban grasses near a major highway in Paris, France, shows that plants store up to 13% of fossil fuel carbon. The ¹⁴C composition of urban grasses is thus a novel parameter to assess the fossil fuel CO₂ pollution.     

Pharmacokinetics of [14C]-atrazine in rhesus monkeys,single-dose intravenous and oral administration

This is the first study regarding the pharmacokinetics of [¹⁴C]-atrazine conducted with rhesus monkeys. The animals received one dose (0.25 mg) intravenously (IV) or three doses (1, 10, or 100 mg) orally. Plasma, urine, and feces were collected at defined times up to 7 days post-dosing. Sample radioactivity was measured to determine the mass equivalent. IV administered [¹⁴C]-atrazine disappeared rapidly from blood, with an elimination half-life of about 5.5 ± 1.1 h. The pharmacokinetic profiles of [¹⁴C]-atrazine following oral administration at the three dose levels show that kinetic parameters such as AUC and C _max are linearly correlated with the dose. Seven days after dosing, urinary and fecal excretion of [¹⁴C]-atrazine reached 99% of total administered dose in the IV group and 91–95% in the three oral dose groups. In the IV-administered monkeys, approximately 85% of the dose was excreted in urine and 12% in feces. In three oral dose groups, urinary and fecal radioactivity recoveries approximated 57% and 21%, 58% and 25%, and 53% and 35%, respectively. More than 50% of the total urinary excreted radioactivity was found within the first 24 h after dosing. In conclusion, the principal elimination of [¹⁴C]-atrazine, IV and orally administered, is via urine. The oral bioavailability was 60% or higher. There was a significant linear correlation between administered oral dose and plasma concentration. Overall oral dose accountability ranged from 91% to 95%. Data generated may be useful in the risk assessment of human exposure to environmental atrazine contamination.     

Variations in the progress of 14C uptake as a source of error in estimates of primary production

The progress curves of ¹⁴C retention for samples of phytoplankton from the Irish Sea incubated at contrasting light intensities have been obtained by two methods. The first method (A) involved the incubation of the samples for various periods up to 6 h, while the second method (B) consisted of making a series of short-term incubations over the same 6 h period. Over this period, the cumulative uptake was tenerally less when estimated by Method A than by Method B. The difference was greater in the samples incubated at the lower light intensity, the light history of the samples having no effect on the difference. The differences has a kinetic basis, with two combinations of progress curves obtained by use of the two methods. The first combination was associated with samples collected in the early morning, while the second combination was exhibited by samples taken in the afternoon irrespective of sampling depth. In certain samples, no increase in the ¹⁴C retained by the cells as measured by Method A was observed after 4 h. The cumulative retention of ¹⁴C by the cells after 2 h was generally greater when estimated by Method A than by Method B, this situation being reversed after 4 h. This reversal indicated a change in uptake kinetics between 2 and 4 h and it is suggested that this interval represents the time necessary for the ¹⁴C to work through the cellular pool of carbon. The findings are discussed in relation to the methodology for obtaining both estimates of primary production and ¹⁴C uptake-light intensity curves for marine phytoplankton.     

Turnover of tracer (14C, 3H labelled) alanine in inshore marine sediments

The turnover rate constants (k) of alanine in coastal marine sediments were measured using ³H-and ¹⁴C-alanine in tracer amounts (less than in situ concentrations of alanine in the porewater). After incubation ¹⁴C-label was recovered in free dissolved alanine, adsorbed alanine, volatile fatty acids and carbon dioxide pools. Alanine left the free dissolved pool by two processes: 1) adsorption (k=0.06 min^-1) and 2) biological uptake (k=0.16 min^-1). Adsorption of alanine was deduced from the persistence of ¹⁴C-alanine, which was slowly metabolized after an initial rapid rate of degradation. Adsorption was confirmed in biologically inactivated sediments. The adsorbed ¹⁴C-alanine was only partially exchanged by the addition of excess non-labelled alanine, indicating the existence of at least 2 different adsorbed pools. The rates of adsorption and desorption were equal, but the k-values were different, indicating that the adsorbed pool was 2500 fold greater than the free pool. From the biological turnover rate constant (0.16 min^-1), the mineralization percentage (80%) and the concentration of alanine (800 nmol l^-1 of porewater) a mineralization rate in the sediment of 75 nmol cm^-3 d^-1 was determined. This was in excess of the measured total NH ₄ ⁺ production. It is concluded that much of the dissolved alanine (800 nm) was biologically unavailable and a more realistic free dissolved pool would be 10 nM.     

Uptake and metabolism of [<Superscript>14</Superscript>C]rinderine and [<Superscript>14</Superscript>C]retronecine in leaf-beetles of the genus <Emphasis Type="Italic"> Platyphora</Emphasis> and alkaloid accumulation in the exocrine defensive secretions

Summary. Sequestration and processing of pyrrolizidine alkaloids (PAs) by leaf beetles of the genus Platyphora were investigated. Tracer experiments with labeled alkaloids were performed with P. eucosma feeding on Koanophyllon panamense (Asteraceae, tribe Eupatorieae). P. eucosma catalyzes the same reactions previously demonstrated for P. boucardi specialized to Prestonia portobellensis (Apocynaceae): (i) epimerization of rinderine to intermedine; (ii) esterification of retronecine yielding insect-specific PAs; (iii) efficient transport of the PAs as free bases into the defensive secretions. P. bella feeding on Tournefortia cuspidata (Boraginaceae) shows the same sequestration behavior and ability to synthesize the specific retronecine esters. P. ligata, a species phylogenetically closely related to the PA adapted species and clustering in the same clade, but feeding on a host plant devoid of PAs, feeds easily on PA treated host-plant leaves, but does not sequester or metabolize PAs. P. kollari a species clustering outside the PA clade refused to feed on its food-plant leaves painted with PAs. The results are discussed in relation to host-plant selection of the PA adapted species and the role of PAs in chemical defense. Received 20 September 2002; accepted 18 November 2002.     

Transfer of the insecticide [<Superscript>14</Superscript>C] imidacloprid from soil to tomato plants

The fate of imidacloprid was investigated in tomato plants during 75 days in soil contaminated by ¹⁴C-imidacloprid. Leaves and fruits were separately analysed for total radioactivity and metabolites. Almost 85% of plant radioactivity was translocated to shoots. Radioactivity concentrations decreased from bottom leaves to top leaves. Desnitro-imidacloprid was the main metabolite in leaves. Nevertheless, more than 50% of the leave radioactivity corresponded to imidacloprid. Residue concentrations were similar in all fruits (62.9 ng g⁻¹), irrespective of their position on plant. In fruits more than 85% of the radioactivity was due to imidacloprid. The small fraction of residues translocated to fruits depended on the low xylem flow in fruits.     

Enhancing the efficiency of nitrogen removing bacterial population to a wide range of C:N ratio (1.5:1 to 14:1) for simultaneous C & N removal

• Simultaneous C & N removal in Methammox occurs at wide C:N ratio. • Biological Nitrogen Removal at wide C:N ratio of 1.5:1 to 14:1 is not reported. • Ammonia removal shifted from mixotrophy to heterotrophy at high C:N ratio. • Acetogenic population compensated for ammonia oxidizers at high C:N ratio. • Methanogens increase the plasticity of nitrogen removers at high C:N ratio. High C:N ratio in the wastewater limits biological nitrogen removal (BNR), especially in anammox based technologies. The present study attempts to improve the COD tolerance of the BNR process by associating methanogens with nitrogen removing bacterial (NRB) populations. The new microbial system coined as ‘Methammox’, was investigated for simultaneous removal of COD (C) and ammonia (N) at C:N ratio 1.5:1 to 14:1. The ammonia removal rate (11.5 mg N/g VSS/d) and the COD removal rates (70.6 mg O/g VSS/d) of Methammox was close to that of the NRB (11.1 mg N/g VSS/d) and the methanogenic populations (77.9 mg O/g VSS/d), respectively. The activities established that these two populations existed simultaneously and independently in ‘Methammox’. Further studies in biofilm reactor fetched a balanced COD and ammonia removal (55%–60%) at a low C:N ratio (≤2:1) and high C:N ratio (≥9:1). The population abundance of methanogens was reasonably constant, but the nitrogen removal shifted from mixotrophy to heterotrophy as the C:N ratio shifted from low (C:N≤2:1) to high (C:N≥9:1). The reduced autotrophic NRB (ammonia- and nitrite-oxidizing bacteria and Anammox) population at a high C:N ratio was compensated by the fermentative group that could carry out denitrification heterotrophically. The functional plasticity of the Methammox system to adjust to a broad C:N ratio opens new frontiers in biological nitrogen removal of high COD containing wastewaters.     

Anreicherung,verteilung, umwandlung und ausscheidung von DDT-14C bei Solea solea (Pisces: Soleidae)

Common soles Solea solea (L.) are extremely suitable for studying the fate of pesticides in marine fish by means of laboratory experiments involving small-sized, accurately controlled, closed aquarium systems. In 5 different experiments, a total of 16 fishes of Age Group I were maintained in water of 10°C and 20‰ S continuously filtered through charcoal, and given oral doses of 0.85 μg DDT-¹⁴C up to 5 times/week. A total application of 1.7, 17 or 35 μg DDT-¹⁴C, corresponding to 3, 28, or 57 experimental days, resulted in 72, 60 or 43% DDT-¹⁴C accumulation. After feeding with 17 μg unlabelled DDT over a period of 4 weeks and final feeding with 1.7 μg ¹⁴C-labelled DDT for 2 days, S. solea displayed the same percentage (74%) of accumulated DDT-¹⁴C as after feeding with 1.7 μg DDT-¹⁴C without prior feeding. Therefore, the decreasing accumulation percentages with inereasing doses, i.e., with longer application periods, are due to elimination during the application period. During a period of 2 months in pesticide-free water, S. solea eliminated 62% of the DDT-¹⁴C which it had accumulated after feeding with 17 μg DDT-¹⁴C over a period of 4 weeks. The gastro-intestinal tract is assumed to be a major route of DDT elimination. Independent of dosage, there was a characteristic distribution pattern of accumulated DDT: brain, liver and gastro-intestinal tract ranked highest, while the concentration in skeletal muscle was lowest. Even during the elimination period the pattern appeared unchanged. DDE, DDD and a polar component occurred as metabolites, but in all organs more than 80% of the accumulated DDT remained unchanged. Percentages of metabolites were higher in liver and gastro-intestinal tract than in skeletal muscle. Prolonged exposure to DDT in the diet induced DDT transformation in the gastro-intestinal contents, most probably in the bacterial flora of the gut.     

Radio (<Superscript>14</Superscript>C)- and fluorescent-doubly labeled silica nanoparticles for biological and environmental toxicity assessment

A new and efficient synthetic route to fluorescent and ¹⁴C-double-labeled silica-based nanoparticles (NPs) is described. The synthesis has been carried out using the “oil-in-water” micro-emulsion technique. Fluorescent and radioactive labeling have been achieved entrapping labeled poly(ethylene glycol) (PEG) molecules in the NPs. The produced particles have been analyzed by means of scanning electron microscopy, photon correlation spectroscopy, confocal microscopy, scintillation counting and oxidation/combustion experiments. Fluorescence quenching experiments confirm that the label is entrapped in the particles. The results presented suggest that the silica matrix does not block the β-radiations emitted from the labeled PEG molecules entrapped in the NPs.     

Dual <Superscript>14</Superscript>C/residue analysis method to assess the microbial accessibility of native phenanthrene in environmental samples

The aim of this work was to develop a method to assess the microbial accessibility of native phenanthrene present in soils and sediments. We developed an accelerated biodegradation assay, characterized by (a) inoculation with a sufficient number of phenanthrene-degrading microorganisms, (b) monitoring of the biodegradation activity through ¹⁴C-mineralization measurements, and (c) single-step chemical analysis of the native compound in the residue. The use of ¹⁴C-labeling allowed the determination of the time period needed for biodegradation of the bioaccessible fraction of the native chemical. The method was tested with environmental samples having a wide range of phenanthrene concentrations, i.e., from background levels (μg kg^-1) originating in soil from atmospheric deposition, to acute concentrations (g kg^-1) corresponding to industrial pollution of soils and sediments. The results showed a wide range of bioaccessibility (15–95% of the initial amount). The method can be used for the assessment of bioaccessibility involved in the management of polycyclic aromatic hydrocarbon (PAH) pollution.     

Fish-bone oil: Percent total body lipid and carbon-14 uptake following feeding of 1-14C-palmitic acid

Bones contain the majority of body lipid in some marine fish. In the sheepshead wrasse Pimelometopon pulchrum and the sablefish Anoplopoma fimbria, the bone lipid comprised 79 to 93% and 52 to 82% of the total body lipid, respectively. The senorita Oxyjulus california, another species of wrasse, has only 14% of its body lipid in bone. To determine whether dietary lipid is deposited quickly in the bone lipid, three species of fish were fed 1-¹⁴C-palmitic acid. Radioactivity appeared in the bone lipid as soon as 12 h after feeding, with the species rich in bone lipid incorporating the highest activity relative to the flesh. Roughly 80% of the radioactivity recovered in A. fimbria bone lipid was found in triglyceride. Radioactivity was equally distributed between phospholipid and triglyceride in P. pulchrum bone lipids. The results suggest that in some marine fish the bones contain the majority of the organism's reserve energy.     

Comparison between the carbon-14 and oxygen consumption method for the determination of the activity of heterotrophic bacterial populations

The activity of the heterotrophic microbial population in the saline Lake Grevelingen (The Netherlands) and the Mediterranean Etang Salses Leucate (France) was determined by measuring the oxygen consumption rate, and the uptake of ¹⁴C-labelled glycollate, pyruvate and an amino acid mixture. The maximum uptake rate of the applied organic compounds in Lake Grevelingen was generally less than 10% of the carbon mineralization rate calculated from the oxygen consumption experiments. Only for pyruvate and glycollate higher values were found of about 30 to 40% with one exceptionally high value for pyruvate of 149%. However, these higher percentages were found in winter, when the activity of the heterotrophic microbial population was very low. In Etang Salses Leucate higher maximum uptake rates of the ¹⁴C-labelled compounds were found, relating this uptake to the oxygen consumption rate. Yet the maximum uptake rate is still always lower than 35% of the carbon mineralization calculated from the oxygen uptake rate. Taking into account that maximum uptake rates were considered, the results demonstrate that the uptake of ¹⁴C-labelled organic compounds represents a serious underestimation of the activity of the bacterial population in situ. The extent of the underestimation depends on the water type. It was concluded that the determination of the heterotrophic activity by measuring oxygen consumption rates offers a better insight into the carbon mineralization process in natural waters than the uptake experiments with ¹⁴C-labelled substrates.Communication no. 228 of the Delta Institute for Hydrobiological Research, Yerseke, The Netherlands     

Long-distance transport of 14C-labelled assimilates in the Fucales: directionality,pathway and velocity

A directed translocation of ¹⁴C-labelled photoassimilates was shown to occur in the thalli of six species of brown seaweeds belonging to the Fucales. The apical growing regions of these algae act as strong sinks in the source-tosink relationship of the long-distance transport. Physical interruptions of the thalli showed that the midrib is the main pathway of the assimilates in Fucus vesiculosus and F. serratus. ¹⁴C-histoautoradiography has shown that translocation takes place through the filamentous cells of the medulla. The translocation velocity of the labelled assimilates, calculated from time source profiles of the tracer, was estimated to be 2 to 4 cm · h^-1. It is suggested that the sieve elements of the Fucales act in a way similar to the conducting sieve elements of the Laminariales.