Radioactive tracer techniques as an aid in resolving sedimentation problems /

Mode of access: Internet.

Application of radiochemistry techniques in food processing research : radioisotopic tracer techniques in evaluation and improvement of industry practices for removal of pesticide residues from foods : annual report no. 4 /

"D-1548."

Application of radiochemistry techniques in food processing research : radioisotopic tracer techniques in evaluation and improvement of industry practices for removal of pesticide residues from foods : annual report /

"TID-4500."

Application of radiochemistry techniques in food processing research : radioisotopic tracer techniques in evaluation and improvement of industry practices for removal of pesticide residues from foods /

Report prepared by project team and edited by Walter A. Mercer.

Quantifying nitrogen cycling rates in freshwater marshes of the southern Everglades using nitrogen-15 tracer techniques

This dissertation research project addressed the question of how hydrologic restoration of the Everglades is impacting the nutrient dynamics of marsh ecosystems in the southern Everglades. These effects were analyzed by quantifying nitrogen (N) cycle dynamics in the region. I utilized stable isotope tracer techniques to investigate nitrogen uptake and cycling between the major ecosystem components of the freshwater marsh system. I recorded the natural isotopic signatures (δ15N and δ 13C) for major ecosystem components from the three major watersheds of the Everglades: Shark River Slough, Taylor Slough, and C-111 basin. Analysis of δ15 N and δ13C natural abundance data were used to demonstrate the spatial extent to which nitrogen from anthropogenic or naturally enriched sources is entering the marshes of the Everglades. In addition, I measured the fluxes on N between various ecosystem components at both near-canal and estuarine ecotone locations. Lastly, I investigated the effect of three phosphorus load treatments (0.00 mg P m-2, 6.66 mg P m-2, and 66.6 mg P m-2) on the rate and magnitude of ecosystem N-uptake and N-cycling. The δ15N and δ13C natural abundance data supported the hypothesis that ecosystem components from near-canal sites have heavier, more enriched δ 15N isotopic signatures than downstream sites. The natural abundance data also showed that the marshes of the southern Everglades are acting as a sink for isotopically heavier, canal-borne dissolved inorganic nitrogen (DIN) and a source for "new" marsh derived dissolved organic nitrogen (DON). In addition, the 15N mesocosm data showed the rapid assimilation of the 15N tracer by the periphyton component and the delayed N uptake by soil and macrophyte components in the southern Everglades.

Deposit-feeding in benthic macrofauna : Tracer studies from the Baltic Sea

A low content of organic matter, which is largely refractory in nature, is characteristic of most sediments, meaning that aquatic deposit-feeders live on a very poor food source. The food is derived mainly from sedimenting phytodetritus, and in temperate waters like the Baltic Sea, from seasonal phytoplankton blooms. Deposit-feeders are either bulk-feeders, or selective feeders, which preferentially ingest the more organic-rich particles in the sediment, including phytodetritus, microbes and meiofauna. The soft-bottom benthos of the Baltic Sea has low species biodiversity and is dominated by a few macrobenthic species, among which the most numerous are the two deposit-feeding amphipods Monoporeia affinis and Pontoporeia femorata, and the bivalve Macoma balthica. This thesis is based on laboratory experiments on the feeding of these three species, and on the priapulid Halicryptus spinulosus. Feeding by benthic animals is often difficult to observe, but can be effectively studied by the use of tracers. Here we used the radioactive isotope 14C to label food items and to trace the organic matter uptake in the animals, while the stable isotopes 13C and 15N were used to follow feeding on aged organic matter in the sediment. The abundance of M. balthica and the amphipods tends to be negatively correlated, i.e., fewer bivalves are found at sites with dense populations of amphipods, with the known explanation that newly settled M. balthica spat are killed by the amphipods. Whether the postlarvae are just accidentally killed, or also ingested after being killed was tested by labelling the postlarvae with 14C and Rhodamine B. Both tracer techniques gave similar evidence for predation on and ingestion of postlarval bivalves. We calculated that this predation was likely to supply less than one percent of the daily carbon requirement for M. affinis, but might nevertheless be an important factor limiting recruitment of M. balthica. The two amphipods M. affinis and P. femorata are partly vertically segregated in the sediment, but whether they also feed at different depths was unknown. By adding fresh 14C-labelled algae either on the sediment surface or mixed into the sediment, we were able to distinguish surface from subsurface feeding. We found M. affinis and P. femorata to be surface and subsurface deposit-feeders, respectively. Whether the amphipods also feed on old organic matter, was studied by adding fresh 14C-labelled algae on the sediment surface, and using aged, one-year-old 13C- and 15N-labelled sediment as deep sediment. Ingestion of old organic matter, traced by the stable isotopes, differed between the two species, with a higher uptake for P. femorata, suggesting that P. femorata utilises the older, deeper-buried organic matter to a greater extent. Feeding studies with juveniles of both M. affinis and P. femorata had not been done previously. In an experiment with the same procedure and treatments as for the adults, juveniles of both amphipod species were found to have similar feeding strategies. They fed on both fresh and old sediment, with no partitioning of food resources, making them likely to be competitors for the same food resource. Oxygen deficiency has become more wide-spread in the Baltic Sea proper in the last half-century, and upwards of 70 000km2 are now devoid of macrofauna, even though part of that area does not have oxygen concentrations low enough to directly kill the macrofauna. We made week-long experiments on the rate of feeding on 14C-labelled diatoms spread on the sediment surface in different oxygen concentrations for both the amphipod species, M. balthica and H. spinulosus. The amphipods were the most sensitive to oxygen deficiency and showed reduced feeding and lower survival at low oxygen concentrations. M. balthica showed reduced feeding at the lowest oxygen concentration, but no mortality increase. The survival of H. spinulosus was unaffected, but it did not feed, showing that it is not a surface deposit-feeder. We conclude that low oxygen concentrations that are not directly lethal, but reduce food intake, may lead to starvation and death in the longer term.

Relationships between nitrogen and phosphorus forms and ratios and the development of dinoflagellate blooms in the East China Sea

During late spring and early summer of 2005, large-scale (> 15 000 km(2)), mixed dinoflagellate blooms developed along the the coast of the East China Sea. Karenia mikimotoi was the dominant harmful algal bloom species in the first stage of the bloom (late May) and was succeeded by Prorocentrum donghaiense approximately 2 wk later. Samples were collected from different stations along both north-south and west-east transects, from the Changjiang River estuary to the south Zhejiang coast, during 3 cruises of the Chinese Ecology and Oceanography of Harmful Algal Blooms Program, before and during the bloom progression. Nitrogen isotope tracer techniques were used to measure rates of NO3-, NH4+, urea, and glycine uptake during the blooms. High inorganic nitrogen (N), but low phosphorus (P) loading from the Changjiang River led to high dissolved inorganic N:dissolved inorganic P ratios in the sampling area and indicate the development of P limitation. The rates of N-15-uptake experiments enriched with PO43- were enhanced compared to unamended samples, suggesting P limitation of the N-uptake rates. The bloom progression was related to the change in availability of both organic and inorganic N and P. Reduced N forms, especially NH4+, were preferentially taken up during the blooms, but different bloom species had different rates of uptake of organic N substrates. K mikimotoi had higher rates of urea uptake, while P. donghaiense had higher rates of glycine uptake. Changes in the availability of reduced N and the ratios of N:P in inorganic and organic forms were suggested to be important in the bloom succession. Nutrient ratios and specific uptake rates of urea were similar when compared to analogous blooms on the West Florida Shelf.

Temporal and spatial variability in nitrogen uptake kinetics during harmful dinoflagellate blooms in the East China Sea

Temporal and spatial variability in the kinetic parameters of uptake of nitrate (NO3-), ammonium (NH4+), urea, and glycine was measured during dinoflagellate blooms in Changjiang River estuary and East China Sea coast, 2005. Karenia mikimotoi was the dominant species in the early stage of the blooms and was succeeded by Prorocentrum donghaiense. The uptake of nitrogen (N) was determined using N-15 tracer techniques. The results of comparison kinetic parameters with ambient nutrients confirmed that different N forms were preferentially taken up during different stages of the bloom. NO3- (V-max 0.044 h(-1); K-s 60.8 mu M-N) was an important N source before it was depleted. NH4+ (V-max 0.049 h(-1); K-s 2.15 mu M-N) was generally the preferred N. Between the 2 organic N sources, urea was more preferred when K. mikimotoi dominated the bloom (V-max 0.020 h(-1); K-s 1.35 mu M-N) and glycine, considered as a dominant amino acid, was more preferred when P. donghaiense dominated the bloom (V-max 0.025 h(-1); K-s 1.76 mu M-N). The change of N uptake preference by the bloom-forming algae was also related to the variation in ambient N concentrations. Published by Elsevier B.V.

SIZE STRUCTURES OF MICROPLANKTON BIOMASS AND PRODUCTION IN JIAOZHOU BAY, CHINA

Seasonal investigations of size-fractionated biomass and production were carried out from February 1992 to May 1993 in Jiaozhou Bay, China. Microplankton assemblages were separated into three fractions: pico-(0.7-2 mu m), nano- (2-20 mu m) and netplankton (20-200 mu m). The biomass was measured as chlorophyll a (Chl a), particulate organic carbon (POC) and particulate organic nitrogen (PON). The production was determined by C-14 and N-15 tracer techniques. The seasonal patterns in biomass, though variable, were characterized by higher values in spring and lower values in autumn and summer (for Chl a only). The seasonal patterns in production, on the other hand, were more clear with higher values occurring in summer and spring, and lower values occurring in autumn and winter. Averaged over the whole study period, the respective proportions of total biomass accounted for by net-, nano- and picoplankton were 26, 45 and 29% for Chl a, 32, 33 and 35% for POC, and 26, 32 and 42% for PON. The contributions to total primary production by net-, nano- and picoplankton were 31, 35 and 34%, respectively. The respective proportions of total NH4+-N uptake accounted for by net-, nano- and picoplankton were 28, 33 and 39% in the daytime, and 10, 29 and 61% at night. The respective contributions to total NO3--N uptake by net-, nano- and picoplankton were 37, 40 and 23% in the daytime, and 13, 23 and 64% at night. Some comprehensive ratios, including C/N biomass ratio, Chl a/C ratio, C uptake/Chl a ratio, C:N uptake ratio and the f-ratio, were also calculated size separately, and their biological and ecological meanings are discussed.

The PRIME 1996 cruise: an overview

The UK PRIME cruise, June-July 1996 in the NE Atlantic, consisted of two legs. During the first, detailed chemical and biological observations were made in time-series mode adjacent to the centre of a cold-core eddy in the vicinity of 59 degreesN 20 degreesW using SF6 tracer techniques as the basis for the Lagrangian study. The eddy, which appeared to have been formed the previous winter, remained coherent over the 9 days of the survey and advected only slowly. The phytoplankton community in the eddy was dominated by the coccolithophorid Coccolithus huxleyi. High microzooplankton grazing rates indicated minimal export losses from the surface layer. Significant shifts in many, but not all, of the chemical and biological properties measured were observed over the course of the experiment, especially after the passage of a storm event, which resulted in considerable deepening of the mixed layer followed by a return to fully stratified conditions. The second leg consisted of a transect from 59 degreesN 20 degreesW to 37 degreesN 19 degreesW, with a further Lagrangian time-series study based on a drogue marker initiated at the southern end of the transect. Maximal biological activity was generally encountered in the region between two fronts located at 52.5 degreesN and 48 degreesN, while to the south of 48 degreesN oligotrophic conditions prevailed. At the southern Lagrangian site, a deep chlorophyll maximum was present and high column new production was recorded as a result of the euphotic zone extending below the depth of the nutricline. Microzooplankton grazing rates were lower at this location than at the northern eddy site. The influx of a warm, saline water body into the upper layers during the southern survey led to a major shift in many of the biological and chemical properties being measured. At both the northern and southern Lagrangian sites, the biomass of the mesozooplankton exceeded that of the microzooplankton. (C) 2001 Elsevier Science Ltd. All rights reserved.

Inhibition of the human epithelial calcium channel TRPV6 by 2-aminoethoxydiphenyl borate (2-APB)

TRPV6, a highly calcium-selective member of the transient receptor potential (TRP) channel superfamily, is a major pathway for calcium absorption in the fetal and adult body. It is expressed abundantly in the duodenum, the placenta and exocrine tissues. TRVP6 was postulated to contribute to store-operated calcium channel (SOC) activity in certain cell types such as exocrine cells. In this study, we tested 2-APB, a widely used SOC inhibitor on human TRPV6 (hTRPV6) activity using fluorescence imaging, patch clamp and radioactive tracer techniques in transiently and stably transfected HEK293 cells. We found that the basal calcium and cadmium influx was higher in HEK293 cells transfected with hTRPV6 than in non-transfected cells. 2-APB inhibited hTRPV6 activity in both transient and stably transfected cells. This effect was slightly sensitive toward extracellular calcium. The extracellular sodium concentration did not affect the inhibition of hTRPV6 by 2-APB. However, N-methyl-d-glucamine significantly diminished the inhibitory effect of 2-APB presumably through direct interaction with this compound. Furthermore, 2-APB inhibited the activity of TRPV6 orthologs but not human TRPV5. 2-APB may serve as a parental compound for the development of therapeutic strategies specifically targeting the hTRPV6 calcium channel.