A microautoradiographic method for fresh-frozen sections to reveal the distribution of radionuclides at the cellular level in plants.

Microautoradiography (MAR) is a conventional imaging method based on the daguerreotype. The technique is used to visualize the distribution of radionuclide-labeled compounds within a tissue section. However, application of the classical MAR method to plant tissue sections is associated with several difficulties. In this study, we report an MAR method applicable to fresh-frozen plant sections. Our method had two features: (i) the sample was kept frozen from plant tissue collection to radioisotope detection, making it possible to fix solutes without solvent exchange; and (ii) 1.2 µm thick polyphenylene sulfide film was inserted between the fresh-frozen plant section and the photosensitive nuclear emulsion to separate the section from the emulsion before autoradiography was conducted, which significantly improved the quality of the section until microscopic detection, the quality of the MAR image and the success rate. Then, the passage of cadmium (Cd) through vegetative rice stem tissue after 24 h of (109)Cd absorption was described for the first time using the MAR method. MAR clearly revealed the distribution of (109)Cd at the tissue level with high resolution. The (109)Cd concentration in phloem cells was found to be particularly high, whereas the xylem cells contained only small amounts of (109)Cd. The MAR method was also applicable for detecting (109)Cd and [(33)P]phosphate in roots. The MAR method developed here is expected to provide distribution images for a variety of compounds and ions in plant tissue.

In situ analysis of cadmium uptake in four sections of the gastro-intestinal tract of rainbow trout (Oncorhynchus mykiss).

This study links results from past in vitro and in vivo experiments, by implementing an in situ experiment in order to determine the relative importance for cadmium (Cd) uptake of different sections of the gastro-intestinal tract (GIT) of rainbow trout. Transport of Cd from four sections of the GIT of adult rainbow trout (~220 g) was individually examined by infusing ligated sections of the GIT in live, free-swimming fish with 50 µM Cd spiked with radiolabelled (109)Cd (0.5 µCi ml(-1)). Fish were exposed for an 8-h period. The percentage of the total injected (109)Cd which was internalized from the different segments was only between ~0.1 and ~7%, indicating low uptake efficiency. The stomach is the most important GIT segment for Cd transport into the internal compartment of the animal, while the posterior intestine also plays a significant role. The majority of (109)Cd recovered at the end of the flux period was detected within gut material (ranging from 28 to 95%); the portion of Cd which was internalized was largely found in the carcass (32 to 60%). Distribution between the measured organs varied with uptake from the various GIT sections. Our results also confirm that the GIT acts as a protective barrier against Cd uptake from dietary exposure.

Real-time imaging and analysis of differences in cadmium dynamics in rice cultivars (Oryza sativa) using positron-emitting 107Cd tracer.

BACKGROUND: Rice is a major source of dietary intake of cadmium (Cd) for populations that consume rice as a staple food. Understanding how Cd is transported into grains through the whole plant body is necessary for reducing rice Cd concentrations to the lowest levels possible, to reduce the associated health risks. In this study, we have visualized and quantitatively analysed the real-time Cd dynamics from roots to grains in typical rice cultivars that differed in grain Cd concentrations. We used positron-emitting 107Cd tracer and an innovative imaging technique, the positron-emitting tracer imaging system (PETIS). In particular, a new method for direct and real-time visualization of the Cd uptake by the roots in the culture was first realized in this work. RESULTS: Imaging and quantitative analyses revealed the different patterns in time-varying curves of Cd amounts in the roots of rice cultivars tested. Three low-Cd accumulating cultivars (japonica type) showed rapid saturation curves, whereas three high-Cd accumulating cultivars (indica type) were characterized by curves with a peak within 30 min after 107Cd supplementation, and a subsequent steep decrease resulting in maintenance of lower Cd concentrations in their roots. This difference in Cd dynamics may be attributable to OsHMA3 transporter protein, which was recently shown to be involved in Cd storage in root vacuoles and not functional in the high-Cd accumulating cultivars. Moreover, the PETIS analyses revealed that the high-Cd accumulating cultivars were characterized by rapid and abundant Cd transfer to the shoots from the roots, a faster transport velocity of Cd to the panicles, and Cd accumulation at high levels in their panicles, passing through the nodal portions of the stems where the highest Cd intensities were observed. CONCLUSIONS: This is the first successful visualization and quantification of the differences in whole-body Cd transport from the roots to the grains of intact plants within rice cultivars that differ in grain Cd concentrations, by using PETIS, a real-time imaging method.

Calibration of (109)Cd KXRF systems for in vivo bone lead measurements: the guiding role of the assumptions for least-squares regression in practical problem solving.

The use of least-squares regression to probe the level of lead contamination of plaster of Paris standards in the calibration of (109)Cd KXRF systems for bone lead measurement, as well as the use of iteratively reweighted least-squares (IRLS) in the case of violation of the assumptions for ordinary least-squares (OLS), is discussed here. One common violation is non-uniform residual variance, which makes the use of OLS inappropriate due to strong influence of points with large variance on the calibration line and variance of the slope and intercept. Comparison between OLS and IRLS in that case showed that IRLS estimates of the intercept are significantly smaller and more precise than OLS estimates, while a less marked increase in the calibration slope is observed when IRLS is used. Moreover, OLS underestimates bone lead concentrations at low levels of lead exposure and overestimates those concentrations at higher levels. These discrepancies are smaller in magnitude than the measurement uncertainty of conventional systems, except for high concentrations. For the newly developed cloverleaf systems, the suggested differences at bone lead concentrations below 17 ppm are comparable to the minimum detection limit, but are larger than the measurement uncertainty for bone lead concentrations above 60 ppm.

Calibration of (109)Cd KXRF systems for in vivo bone lead measurements: weighted least-squares regression with different weighting functions.

The use of iteratively reweighted least squares (IRLS) has recently been described as an alternative to ordinary least squares with heteroscedastic data, in the calibration of (109)Cd KXRF systems for in vivo bone lead measurements. This work addresses the use of weighted least squares (WLS) with two different weighting functions and no iteration, with that same data set. The functions are defined as the inverse of the variance of observed ratios of lead to coherent peak amplitudes and the inverse of the square of the error reported by the Marquardt fitting program for these ratios. The results show that if no iteration is implemented when using WLS, then the two weighting functions are highly inefficient in homogenizing the residual variance. Moreover, both methods estimate much more imprecise calibration intercepts and slopes than did the IRLS method. Work is in progress to investigate the implementation of IRLS with these weighting functions, with the focus on the selection of the best function for residuals to be used in each iteration stage.

Using isotopes to trace freshly applied cadmium through mineral phosphorus fertilization in soil-fertilizer-plant systems.

Applications of mineral phosphorus (P) fertilizer can lead to cadmium (Cd) accumulation in soils and can increase Cd concentrations in edible crop parts. To determine the fate of freshly applied Cd, a Cd source tracing experiment was conducted in three soil-fertilizer-wheat systems by using a mineral P fertilizer labeled with the radio isotope 109Cd and by exploiting natural differences in Cd stable isotope compositions (δ114/110Cd). Source tracing with stable isotopes overestimated the proportion of Cd in plants derived from the P fertilizer, because the isotope ratios of the sources were not sufficiently distinct from those of the soils. Despite indistinguishable extractable Cd pools between control and treatments, the addition of P fertilizer resulted in a more negative apparent isotope fractionation between soil and wheat. Overall, the radio isotope approach provided more robust results and revealed that 6.5 to 15% of the Cd in the shoot derived from the fertilizer. From the introduced Cd, a maximum of 2.2% reached the wheat shoots, whilst 97.8% remained in the roots and soils. The low recoveries of the fertilizer derived Cd suggest that continuous P fertilizer application in the past decades can lead to a build-up of a residual Cd pool in soils.

Comparative accumulation of (109)Cd and (75)Se from water and food by an estuarine fish (Tetractenos glaber).

Few data are available on the comparative accumulation of metal(loid)s from water and food in estuarine/marine fish. Smooth toadfish (Tetractenos glaber), commonly found in estuaries in south-eastern Australia, were separately exposed to radio-labelled seawater (14kBqL(-1) of (109)Cd and 24kBqL(-1) of (75)Se) and food (ghost shrimps; Trypaea australiensis: 875Bqg(-1)(109)Cd and 1130Bqg(-1)(75)Se) for 25 days (uptake phase), followed by exposure to radionuclide-free water or food for 30 days (loss phase). Toadfish accumulated (109)Cd predominantly from water (85%) and (75)Se predominantly from food (62%), although the latter was lower than expected. For both the water and food exposures, (109)Cd was predominantly located in the gut lining (60-75%) at the end of the uptake phase, suggesting that the gut may be the primary pathway of (109)Cd uptake. This may be attributed to toadfish drinking large volumes of water to maintain osmoregulation. By the end of the loss phase, (109)Cd had predominantly shifted to the excretory organs - the liver (81%) in toadfish exposed to radio-labelled food, and in the liver, gills and kidney (82%) of toadfish exposed to radio-labelled water. In contrast, (75)Se was predominantly located in the excretory organs (gills, kidneys and liver; 66-76%) at the end of the uptake phase, irrespective of the exposure pathway, with minimal change in percentage distribution (76-83%) after the loss phase. This study emphasises the importance of differentiating accumulation pathways to better understand metal(loid) transfer dynamics and subsequent toxicity, in aquatic biota.

Effects of ocean acidification on 109Cd, 57Co, and 134Cs bioconcentration by the European oyster (Ostrea edulis): Biokinetics and tissue-to-subcellular partitioning.

The uptake and depuration kinetics of dissolved 109Cd, 57Co and 134Cs were determined experimentally in the European flat oyster Ostrea edulis (Linnaeus, 1758) under different pH conditions (i.e., 8.1, 7.8 and 7.5) for 59 days. Uptake and depuration rates were variable within these elements; no effects were observed under different pH conditions for the uptake biokinetics of 109Cd and 57Co and depuration of 109Cd and 134Cs in oyster. The uptake and depuration rate constants of 134Cs differed during the exposure phase between treatments, while the steady state concentration factors (CFss) were similar. The resulting Cs activity that was purged during short- and long-term depuration phases differed, while the remaining activities after thirty-nine days depuration phase (RA39d) were similar. Co-57 depuration was affected by pCO2 conditions: RA39d were found to be significantly higher in oysters reared in normocapnia (pCO2 = 350 µatm) compared to high pCO2 conditions. Co-57 tissue distribution did not differ among the variable pCO2 conditions, while 109Cd and 134Cs accumulated in soft tissue of oysters were found to be higher under the highest pCO2. Additionally, Cd, Co and Cs were stored differently in various compartments of the oyster cells, i.e. cellular debris, metal-rich granules (MRG) and metallothionein-like proteins (MTLP), respectively. The subcellular sequestration of the elements at the end of the depuration phase did not differ among pH treatments. These results suggest that bioconcentration and tissue/subcellular distribution are element-specific in the oyster, and the effects of higher pCO2 driven acidification and/or coastal acidification variably influence these processes.

Influence of food quality and salinity on dietary cadmium availability in Mytilus trossulus.

Surficial sediments (a combination of re-suspended and suspended sediments denoted as RSS) were collected from two distinct marine intertidal habitats. The two habitats differed with respect to salinity (25ppt versus 15ppt) and RSS % organic carbon content (24% versus 15%). Feeding experiments were conducted simulating the conditions in the two habitats to determine if salinity and RSS % organic carbon content affected cadmium accumulation in the pacific blue mussel Mytilus trossulus. Eleven different treatments including pure phytoplankton, collected RSS and control clay were radiolabeled with (109)Cd and pulse-fed to M. trossulus under both high (25ppt) and low salinities (15ppt). Metal uptake and accumulation was determined using the DYMBAM biodynamic metal bioaccumulation model. Although M. trossulus ingestion rates (IR) were significantly higher at 25ppt as compared to 15ppt, assimilation efficiencies (AEs) and [(109)Cd] tissue levels were significantly lower at high as compared to low salinity exposures. Of the abiotic and biotic parameters examined and in contrast to other studies, differences in salinity rather than ingestion rate or food quality (as defined by % organic carbon content) seemed to best define the observed differences in (109)Cd AE by M. trossulus.

Improvements in the calibration of 109Cd K x-ray fluorescence systems for measuring bone lead in vivo.

A 109Cd K x-ray fluorescence (XRF) system using a point source in a back-scatter geometry is described. The suitability of plaster-of-Paris phantoms as targets for intercalibration standards was evaluated. When the phantom concentrations were measured by inductively-coupled-plasma mass spectrometry (ICPMs), the calculated phantom concentrations underestimated true concentrations by an average of 15%. Since calculated values are used to calibrate the K XRF system, in vivo bone-lead concentrations may be similarly underestimated. The difference between calculated and measured concentration is attributable to impurities in the plaster of Paris (e.g. calcium carbonate). The ICPMS-measured concentrations were used to calibrate the K XRF system. The same phantoms were also measured as 'unknowns' by a bone-lead measurement system (Abiomed, Danvers, MA, USA). The commercial system overestimated the lowest-concentration phantoms and underestimated the phantoms with concentrations above 15 microg Pb/g plaster of Paris. The commercial system and our system were compared by measurement of the new phantoms in air and in water. The K XRF system exhibited better precision in both situations. On the basis of this work, we recommend that plaster-of-Paris phantoms used to calibrate K XRF measurement systems be analysed first by ICPMS or another valid analytical technique.

Remobilisation of 109Cd, 65Zn and 54Mn from freshwater-labelled river sediments when mixed with seawater.

A major fraction of trace metals transported by rivers is associated with sediments, especially during flooding, when erosion and resuspension increase sediment loads. Upon contact with seawater in estuaries, changes in ionic strength and pH may remobilise trace metals from sediment surfaces into more bioavailable forms. The objective of the present work was to investigate time-dependent interactions between trace metals and freshwater sediments and their potential remobilisation upon contact with seawater. Two river sediments (one organic and one inorganic) were labelled with 109Cd2+, 65Zn2+ and 54Mn2+ radioactive tracers for periods up to 6 months. Sorption of tracers occurred rapidly (> or = 80% sorption, < 1 h), followed by a slower approach to pseudoequilibrium. Kd(6 months) were estimated as 460, 480 and 2200 ml/g (inorganic sediment) compared to 5300, 4000 and 1200 ml/g (organic sediment) for 109Cd, 65Zn and 54Mn, respectively. Remobilisation of tracers from labelled sediments was studied using sequential extractions. Artificial seawater extracts simulated an estuarine environment. Subsequent extractions provided information about more strongly sorbed tracer fractions within sediments. Remobilisation of 109Cd by seawater was significant (> 65%) and least affected by sediment type or freshwater labelling time. Redistribution of Cd to strongly bound phases was minimal (4% and 1% of 109Cd in strongly oxidisable fractions). Seawater remobilisation of 65Zn was significantly greater from the organic sediment (54%) compared to the inorganic sediment (8%), where a large fraction of 65Zn (14%) became irreversibly bound. Similarly, more 54Mn was remobilised by seawater from the organic sediment than the inorganic sediment (66% and 3% remobilised, respectively), i.e., 54Mn became more strongly bound in the inorganic sediment. A simple three-box model, based on first-order differential equations, was used to describe the interaction between tracers in spiked freshwater and two operationally defined sediment fractions ("seawater exchangeable" and "seawater unexchangeable") up to 6 months of freshwater labelling. Model simulations were fitted to experiment data and apparent rate constants were calculated using numerical optimisation methods. Sorption ratios from modelling data (i.e., k1/k2) were greater for organic compared to inorganic sediments, while fixation ratios were higher in inorganic sediments. In conclusion, trace metals can be remobilised from sediments on contact with seawater in estuaries. High organic content in sediments increased initial sorption of tracers but inhibited redistribution to more strongly bound fractions over time, resulting in greater remobilisation of tracers when in contact with seawater.

Estimation of apparent rate coefficients for radionuclides interacting with marine sediments from Novaya Zemlya.

To assess the impact of radionuclides entering the marine environment from dumped nuclear waste, information on the physico-chemical forms of radionuclides and their mobility in seawater-sediment systems is essential. Due to interactions with sediment components, sediments may act as a sink, reducing the mobility of radionuclides in seawater. Due to remobilisation, however, contaminated sediments may also act as a potential source of radionuclides to the water phase. In the present work, time-dependent interactions of low molecular mass (LMM, i.e. species < 10 kDa) radionuclides with sediments from the Stepovogo Fjord, Novaya Zemlya and their influence on the distribution coefficients (Kd values) have been studied in tracer experiments using 109Cd2+ and 60Co2+ as gamma tracers. Sorption of the LMM tracers occurred rapidly and the estimated equilibrium Kd(eq)-values for 109Cd and 60Co were 500 and 20000 ml/g, respectively. Remobilisation of 109Cd and 60Co from contaminated sediment fractions as a function of contact time was studied using sequential extraction procedures. Due to redistribution, the reversibly bound fraction of the gamma tracers decreased with time, while the irreversibly (or slowly reversibly) associated fraction of the gamma tracers increased. Two different three-compartment models, one consecutive and one parallel, were applied to describe the time-dependent interaction of the LMM tracers with operationally defined reversible and irreversible (or slowly reversible) sediment fractions. The interactions between these fractions were described using first order differential equations. By fitting the models to the experimental data, apparent rate constants were obtained using numerical optimisation software. The model optimisations showed that the interactions of LMM 60Co were well described by the consecutive model, while the parallel model was more suitable to describe the interactions of LMM 109Cd with the sediments, when the squared sum of residuals were compared. The rate of sorption of the irreversibly (or slowly reversibly) associated fraction was greater than the rate of desorption of the reversibly bound fractions (i.e. k3 > k2) for both radionuclides. Thus, the Novaya Zemlya sediment are supposed to act as a sink for the radionuclides under oxic conditions, and transport to the water phase should mainly be attributed to resuspended particles.

Radiotracer analysis of cadmium speciation in soil solutions using electrophoresis, dialysis, centrifugation, and ultrafiltration.

Speciation of carrier-free 109Cd, added in cationic form to pre-filtered extracts obtained by leaching forest soil samples with distilled water, was analyzed using electrophoresis, dialysis, centrifugation, and ultrafiltration. Rapid establishment of isotopic equilibrium between the added 109Cd and stable cadmium present in the extracts was observed. All the data obtained indicated that 109Cd and also stable cadmium were present in the analyzed extracts in the form of neutral or negatively charged organic complexes or small colloids. The results of electrophoresis enabled the characterization, at least semiquantitative, of the abundance and electrophoretic mobility of the forms present. The incomplete dialysis of 109Cd from the soil extracts through cellophane membrane against water proved the presence of organic associates with molecular weights higher than 10(4). Dialysis against the same, but unlabeled extract was always complete, indicating the reversible (labile) nature of the organic forms of cadmium. Assessment of the stability constants of the organic forms using a simple discrete two-site model suggested that humate and/or fulvate complexes of cadmium were formed.

Comprehensive RNAA of cadmium, cobalt, nickel, and copper using 109Cd, 57Co, and reactor-produced 67Cu as radioisotopic yield monitors.

An existing radiochemical NAA procedure for Cd, Co, and Cu was improved to allow determination of individual radiochemical yields by the radioisotopic tracer technique, thus eliminating errors owing to variable recovery. 109Cd was used as tracer for Cd determination via 115Cd/115mIn, 57Co for Co via 60Co, and potentially for Ni via 58Co, whereas as a novelty 67Cu, produced by reactor irradiation of ZnO of natural isotopic composition (by the 67Zn [n,p] 67Cu reaction) was used for Cu via the indicator nuclide 64Cu. The simple production and purification of 67Cu by anion exchange is described. Results for biological RMs are given and discussed.

Standardization of 125I and 109Cd by the photon-photon coincidence method in PTKMR-BATAN.

A photon-photon coincidence system was constructed for the standardization of (125)I and (109)Cd in PTKMR-BATAN, Indonesia. Two NaI(Tl) detectors of 76 mm diameter × 6mm thickness with 0.5mm aluminum window were used, which were positioned approximately symmetrically to the source holder. The electronic chain was almost the same as for a 4πß-γ system. The CANBERRA Multiport II multi channel analyzer was used for energy calibration and a Philips type PM3092 oscilloscope for visualization of the pulses. A polyethylene plastic was used as the source substrate for the (125)I and (109)Cd samples. The activity of a (125)I solution was measured by the photon-photon coincidence and the efficiency extrapolation method (Schrader and Walz, 1987), whereas the activity of a (109)Cd solution was determined by a tracer method using (125)I (Schrader, 2006). The result of the (125)I activity showed good agreement with the result of measurements using a calibrated ionization chamber, and the result of (109)Cd also showed good agreement with the measurements result using a LEGe detector.

Cadmium bioaccumulation and retention kinetics in the Chilean blue mussel Mytilus chilensis: seawater and food exposure pathways.

The Chilean blue mussel (Mytilus chilensis, Hupe 1854) represents the most important bivalve exploited along the Chilean coast and is a major food source for the Chilean population. Unfortunately, local fish and shellfish farming face severe problems as a result of bioaccumulation of toxic trace metals into shellfishes. Blue mussels collected along the Chilean coasts contain levels of Cd above the regulatory limits for human consumption. In this study, we examined the bioaccumulation, depuration and organ distribution of Cd in the M. chilensis, from 109Cd-labelled bulk seawater and from feeding with 109Cd-labelled algae. The uptake of 109Cd via seawater displayed a simple exponential kinetic model suggesting that cadmium activity tends to reach an equilibrium value of 1.838+/-0.175 ng g(-1) (mean+/-asymptotic standard error, p < 0.001) after 78+/-9 days. The depuration rate for 109Cd accumulated via seawater was slow, with only 21% of the total 109Cd accumulated in the whole mussel being eliminated after 52 days. Total elimination of Cd in mussels was adequately described by a double component kinetic model, in which the biological half-life for the long-lived component represents more than 6 months. In contrast, depuration after radiolabelled food uptake was fast, reaching only 20% of retention in 10 days. This knowledge of the long half-life of cadmium accumulated via seawater as well as the non-negligible level of cadmium accumulated into the shells is relevant to the management of Cd levels in this species and the refinement of detoxification processes in order to comply with authorized Cd levels.

The barnacle Balanus amphitrite as a biomonitor for Cd: radiolabelled experiments.

Radiolabelled experiments were carried out to measure necessary parameters in the development of a biodynamic ecotoxicological simulation model of Cd accumulation in the barnacle biomonitor Balanus amphitrite. The Cd uptake rate constant from the dissolved phase, the Cd assimilation efficiency (AE) from suspended particulate matter (SPM) and the efflux rate constant were obtained using (109)Cd. A Cd uptake rate constant from the dissolved phase (k(u)) of 0.0072 Lg(-1)h(-1) was determined for the barnacle under environmentally realistic dissolved Cd concentrations (maximum of 400 ng L(-1)). Cd AE from SPM was determined from the barnacle feeding on SPM with low and high chl a concentrations, resulting in AEs of 39.0% and 48.7%, respectively, and an efflux rate of 0.0072 d(-1). The difference between the AEs resulted from differences in chl a:SPM ratios suggesting a general tendency of higher AE when SPM is enriched with chl a. These results reinforce that the accuracy of ecotoxicological models for metal accumulation in organisms depends on how representative the selected food items are of the organism's natural diet.

New best estimates for radionuclide solid-liquid distribution coefficients in soils. Part 3: miscellany of radionuclides (Cd, Co, Ni, Zn, I, Se, Sb, Pu, Am, and others).

New best estimates for the solid-liquid distribution coefficient (K(d)) for a set of radionuclides are proposed, based on a selective data search and subsequent calculation of geometric means. The K(d) best estimates are calculated for soils grouped according to the texture and organic matter content. For a limited number of radionuclides this is extended to consider soil cofactors affecting soil-radionuclide interaction, such as pH, organic matter content, and radionuclide chemical speciation. Correlations between main soil properties and radionuclide K(d) are examined to complete the information derived from the best estimates with a rough prediction of K(d) based on soil parameters. Although there are still gaps for many radionuclides, new data from recent studies improve the calculation of K(d) best estimates for a number of radionuclides, such as selenium, antimony, and iodine.

Bioavailability of cadmium from infant diets in newborn rats.

Infants are exposed to higher levels of cadmium (Cd) from infant and follow-on formulas than from breast milk. We studied the bioavailability of 109CdCl2 from cows' milk formula, soy formula, wheat/oat/milk formula, wholemeal/milk formula and water in 11-day-old rat pups. The pups received a single oral dose of one diet labelled with 109Cd, 0.1 or 0.3 mg Cd/kg body weight. After 2 or 24 h or 4, 9 or 12 days the fractional retention of 109Cd in the whole body, in segments of rinsed small intestine and in tissue was measured in a gamma counter. Pups receiving 109Cd in water or cows' milk formula had the highest mean whole-body retention. It ranged from 67% of the dose in the water group to 52% in the wholemeal/milk formula group 4 days after dosing. The retention of 109Cd in the rinsed small intestine was significantly higher in the water group and the cows' milk formula group than in the cereal-based formula groups at 24 h and 4 days after dosing. It was still high in all groups on day 9, ranging from 26 to 11%. Initially most of the 109Cd was retained in the duodenum but by day 4 it had moved further down into the jejunum. In the liver, the highest and lowest retention on day 4 was 16%, and 3 per thousand of the dose in the water group and wholemeal/milk formula group, respectively. In the kidney, 109Cd was still increasing 12 days after exposure in all groups. Whole-body retention and tissue levels were higher than previously reported in adult animals. The lower bioavailability of 109Cd from the cereal-based formulas compared to water and cows milk formula on the longer survival times is most likely explained by Cd binding to dietary fibre and phytic acid in the cereal-based formulas reducing the intestinal binding and decreasing the bioavailability of Cd. The high retention of 109Cd in the small intestine, leading to a prolonged absorption period, emphasizes the importance of extending studies on neonatal Cd absorption over a long time period in order to detect for example, endpoints, accumulation of Cd in the kidney.