Variation in Shrimp Allergens: Place of Origin Effects on Food Safety Assessment.

Due to the widespread use of shellfish ingredients in food products, accurate food labelling is urgently needed for consumers with shellfish allergies. Most crustacean allergen detection systems target the immunorecognition of the allergenic protein tropomyosin. However, this mode of detection may be affected by an origin-dependent protein composition. This study determined if the geographic location of capture, or aquaculture, influenced the allergenic protein profiles of Black Tiger Shrimp (Penaeus monodon), one of the most farmed and consumed shrimp species worldwide. Protein composition was analysed in shrimp from nine different locations in the Asia-Pacific by SDS-PAGE, immunoblotting, and mass spectrometry. Ten of the twelve known shrimp allergens were detected, but with considerable differences between locations. Sarcoplasmic calcium-binding protein, myosin light chain, and tropomyosin were the most abundant allergens in all locations. Hemocyanin-specific antibodies could identify up to six different isoforms, depending on the location of origin. Similarly, tropomyosin abundance varied by up to 13 times between locations. These findings suggest that allergen abundance may be related to shrimp origin and, thus, shrimp origin might directly impact the readout of commercial crustacean allergen detection kits, most of which target tropomyosin, and this should be considered in food safety assessments.

Evaluating the Application of Portable Handheld X-ray Fluorescence (XRF) Scanner for Determining Seafood Provenance: A Case Study on Penaeus monodon.

Seafood elemental profiling (SEP) is the quantification of a range of elements in seafood products and may serve in addressing questions of seafood provenance and quality. Traditional methods for analyzing soft tissue present several limitations for the industry-level use of SEP. Portable handheld X-ray fluorescence (pXRF) analysis is a promising alternative to conventional methods; however, its application for biological analysis has not been fully established. Intact giant tiger prawn (Penaeus monodon) abdomens were analyzed with a Vanta M series XRF portable analyzer following a novel soft tissue protocol. Exploratory statistics (principal component analysis, nonmetric multidimensional scaling, and canonical discriminant analysis), as well as random forest models, have been implemented with pXRF profiles, yielding 81% accuracy when assigning the geographical origin of P. monodon. The results of this study highlight that SEP via pXRF is a viable industry-level analysis, and its application will depend on improved instrument calibration to account for fluctuating wetness factors that are influenced by cooking, storage, and other pre- and post-harvest treatments.

Developing a MySQL Database for the Provenance of Black Tiger Prawns (Penaeus monodon).

As the demand for seafood increases, so does the incidence of seafood fraud. Confirming provenance of seafood is important to combat fraudulent labelling but requires a database that contains the isotopic and elemental "fingerprints" of authentic seafood samples. Local isotopic and elemental databases can be scaled up or combined with other databases to increase the spatial and species coverage to create a larger database. This study showcases the use of isotopic and elemental fingerprints of the black tiger prawn (Penaeus monodon) to develop a database that can be used to securely store the data necessary for determining provenance. The utility of this database was tested through querying and building seven different datasets that were used to develop models to determine the provenance of P. monodon. The models built using the data retrieved from the database demonstrated that the provenance of P. monodon could be determined with >80% accuracy. As the database was developed using MySQL, it can be scaled up to include additional regions, species, or methodologies depending on the needs of the users. Combining the database with methods of determining provenance will provide regulatory bodies and the seafood industry with another provenance tool to combat fraudulent seafood labelling.

Tracking the provenance of octopus using isotopic and multi-elemental analysis.

Octopus play an increasingly important role in ocean ecosystems and global fisheries, yet techniques for authenticating provenance are sorely lacking. For the first time, we investigate whether chemical profiling can distinguish geographical origins of octopus on international and domestic scales. Our samples consisted of wild-caught octopus from south-east Asia and southern Australia, regions with high seafood trade. We used a novel combination of stable carbon (δ13C) and oxygen (δ18O) isotope analyses (Isotope-Ratio Mass Spectrometry) of internal calcified structures called statoliths, with elemental analyses (X-Ray Fluorescence using Itrax) of soft-tissue. We found that multivariate profiles exhibited distinctive regional signatures, even across species, with high classification success (∼95%) back to region of origin. This study validates isotopic and multi-elemental profiling as an effective provenance tool for octopus, which could be used to support transparency and accountability of seafood supply chains and thus encourage sustainable use of ocean resources.

Tracking the impacts of nutrient inputs on estuary ecosystem function.

Estuaries are one of the most impacted coastal environments globally, subjected to multiple stressors from urban, industry and coastal development. With increasing anthropogenic activity surrounding estuarine systems, sewage inputs have become a common concern. Stable isotope analysis provides a well-established tool to investigate the incorporation of nitrogen into marine organisms and identify major nutrient sources. Benthic macroinvertebrate communities are often used as bioindicators in ecological studies as they typically display predictable responses to anthropogenic pressures, however have a suite of limitations and costs associated with their use. 16S rDNA amplicon sequencing techniques allow for investigation of the microbial communities inhabiting complex environmental samples, with potential as a tool in the ecological assessment of pollution. These communities have not yet been adequately considered for ecological studies and biomonitoring, with a need to better understand interactions with environmental stressors and implications for ecosystem function. This study used a combination of stable isotope analysis to trace the uptake of anthropogenic nitrogen in biota, traditional assessment of benthic macroinvertebrate communities, and 16S rDNA genotyping of benthic microbial communities. Stable isotope analysis of seagrass and epiphytes identified multiple treated and untreated sewage inputs, ranges of 5.2-7.2‰ and 1.9-4.0‰ for δ15N respectively, as the dominant nitrogen source at specific locations. The benthic macroinvertebrate community reflected these inputs with shifts in dominant taxa and high abundances of polychaetes at some sites. Microbial communities provided a sensitive indication of impact with a breadth of information not available using traditional techniques. Composition and predicted function reflected sewage inputs, particularly within sediments, with the relative abundance of specific taxa and putative pathogens linked to these inputs. This research supports the growing body of evidence that benthic microbial communities respond rapidly to anthropogenic stressors and have potential as a monitoring tool in urban estuarine systems.

Correction: Diet type influences the gut microbiome and nutrient assimilation of Genetically Improved Farmed Tilapia (Oreochromis niloticus).

[This corrects the article DOI: 10.1371/journal.pone.0237775.].

Rainfall as a trigger of ecological cascade effects in an Australian groundwater ecosystem.

Groundwaters host vital resources playing a key role in the near future. Subterranean fauna and microbes are crucial in regulating organic cycles in environments characterized by low energy and scarce carbon availability. However, our knowledge about the functioning of groundwater ecosystems is limited, despite being increasingly exposed to anthropic impacts and climate change-related processes. In this work we apply novel biochemical and genetic techniques to investigate the ecological dynamics of an Australian calcrete under two contrasting rainfall periods (LR-low rainfall and HR-high rainfall). Our results indicate that the microbial gut community of copepods and amphipods experienced a shift in taxonomic diversity and predicted organic functional metabolic pathways during HR. The HR regime triggered a cascade effect driven by microbes (OM processors) and exploited by copepods and amphipods (primary and secondary consumers), which was finally transferred to the aquatic beetles (top predators). Our findings highlight that rainfall triggers ecological shifts towards more deterministic dynamics, revealing a complex web of interactions in seemingly simple environmental settings. Here we show how a combined isotopic-molecular approach can untangle the mechanisms shaping a calcrete community. This design will help manage and preserve one of the most vital but underrated ecosystems worldwide.

Refining trophic dynamics through multi-factor Bayesian mixing models: A case study of subterranean beetles.

Food web dynamics are vital in shaping the functional ecology of ecosystems. However, trophic ecology is still in its infancy in groundwater ecosystems due to the cryptic nature of these environments. To unravel trophic interactions between subterranean biota, we applied an interdisciplinary Bayesian mixing model design (multi-factor BMM) based on the integration of faunal C and N bulk tissue stable isotope data (δ13C and δ15N) with radiocarbon data (Δ14C), and prior information from metagenomic analyses. We further compared outcomes from multi-factor BMM with a conventional isotope double proxy mixing model (SIA BMM), triple proxy (δ13C, δ15N, and Δ14C, multi-proxy BMM), and double proxy combined with DNA prior information (SIA + DNA BMM) designs. Three species of subterranean beetles (Paroster macrosturtensis, Paroster mesosturtensis, and Paroster microsturtensis) and their main prey items Chiltoniidae amphipods (AM1: Scutachiltonia axfordi and AM2: Yilgarniella sturtensis), cyclopoids and harpacticoids from a calcrete in Western Australia were targeted. Diet estimations from stable isotope only models (SIA BMM) indicated homogeneous patterns with modest preferences for amphipods as prey items. Multi-proxy BMM suggested increased-and species-specific-predatory pressures on amphipods coupled with high rates of scavenging/predation on sister species. SIA + DNA BMM showed marked preferences for amphipods AM1 and AM2, and reduced interspecific scavenging/predation on Paroster species. Multi-factorial BMM revealed the most precise estimations (lower overall SD and very marginal beetles' interspecific interactions), indicating consistent preferences for amphipods AM1 in all the beetles' diets. Incorporation of genetic priors allowed crucial refining of the feeding preferences, while integration of more expensive radiocarbon data as a third proxy (when combined with genetic data) produced more precise outcomes but close dietary reconstruction to that from SIA + DNA BMM. Further multidisciplinary modeling from other groundwater environments will help elucidate the potential behind these designs and bring light to the feeding ecology of one the most vital ecosystems worldwide.

Diet type influences the gut microbiome and nutrient assimilation of Genetically Improved Farmed Tilapia (Oreochromis niloticus).

Nile tilapia, Oreochromis niloticus is the third most commonly farmed finfish species in the world, accounting for nearly 5% of global aquaculture production. In the past few decades much of the success of this species has been attributed to the development and distribution of Genetically Improved Farmed Tilapia (GIFT). Despite the increasing availability of GIFT, the productivity of small-scale farming remains highly variable, particularly in developing nations. Commercial fish-feed pellets can increase fish farm productivity; however, many small-scale farmers rely on other means of feeding fish due to the high cost and limited availability of commercial fish feed pellets. Therefore, understanding how locally-sourced feeds affect the production of GIFT is an important step towards improving feeding practices, particularly for farmers with low financial capital. This study used stable isotope analysis (SIA) and 16S rRNA gene sequencing to compare the effects of a locally-sourced vegetable-based diet and commercial pellet-based diets on the relative condition, nutrient assimilation patterns and gastrointestinal microbiota of GIFT. GIFT fed a locally-sourced diet were smaller, and in a significantly poorer condition than those fed with commercial fish feeds. SIA showed no differences in dietary carbon between the two diets; however, δ13C, poor fish condition and the abundance of specific bacterial taxa (of such as Fusobacteria) were correlated. SIA revealed that GIFT fed locally-sourced diets that predominantly consisted of vegetables were significantly enriched in δ15N despite a perceived lack of dietary protein. This enrichment suggests that GIFT fed a locally-sourced diet may be supplementing their diet via cannibalism, a behaviour representative of poor farming practice. Overall this study highlights the need to increase the availability of suitable GIFT feeds in developing nations. The development a low-cost feed alternative could improve the success of small-scale GIFT farmers in PNG, increasing both food and income security within the region.

Elucidating stygofaunal trophic web interactions via isotopic ecology.

Subterranean ecosystems host highly adapted aquatic invertebrate biota which play a key role in sustaining groundwater ecological functioning and hydrological dynamics. However, functional biodiversity studies in groundwater environments, the main source of unfrozen freshwater on Earth, are scarce, probably due to the cryptic nature of the systems. To address this, we investigate groundwater trophic ecology via stable isotope analysis, employing δ13C and δ15N in bulk tissues, and amino acids. Specimens were collected from a shallow calcrete aquifer in the arid Yilgarn region of Western Australia: a well-known hot-spot for stygofaunal biodiversity. Sampling campaigns were carried out during dry (low rainfall: LR) and the wet (high rainfall: HR) periods. δ13C values indicate that most of the stygofauna shifted towards more 13C-depleted carbon sources under HR, suggesting a preference for fresher organic matter. Conversion of δ15N values in glutamic acid and phenylalanine to a trophic index showed broadly stable trophic levels with organisms clustering as low-level secondary consumers. However, mixing models indicate that HR conditions trigger changes in dietary preferences, with increasing predation of amphipods by beetle larvae. Overall, stygofauna showed a tendency towards opportunistic and omnivorous habits-typical of an ecologically tolerant community-shaped by bottom-up controls linked with changes in carbon flows. This study provides baseline biochemical and ecological data for stygofaunal trophic interactions in calcretes. Further studies on the carbon inputs and taxa-specific physiology will help refine the interpretation of the energy flows shaping biodiversity in groundwaters. This will aid understanding of groundwater ecosystem functioning and allow modelling of the impact of future climate change factors such as aridification.

Macro-charcoal accumulation in floodplain wetlands: Problems and prospects for reconstruction of fire regimes and environmental conditions.

Floodplain wetland ecosystems respond dynamically to flooding, fire and geomorphological processes. We employed a combined geomorphological and environmental proxy approach to assess allochthonous and autochthonous macro-charcoal accumulation in the Macquarie Marshes, Australia, with implications for the reconstruction of fire regimes and environmental conditions in large, open-system wetlands. After accounting for fluvial macro-charcoal flux (1.05 ± 0.32 no. cm-2 a-1), autochthonous macro-charcoal in ~1 m deep sediment profiles spanning ~1.7 ka were highly variable and inconsistent between cores and wetlands (concentrations from 0 to 438 no. cm-3, mean accumulation rates from 0 to 3.86 no. cm-2 a-1). A positive correlation existed between the number of recent fires, satellite-observed ignition points, and macro-charcoal concentrations at the surface of the wetlands. Sedimentology, geochemistry, and carbon stable isotopes (δ13C range -15 to -25 ‰) were similar in all cores from both wetlands and varied little with depth. Application of macro-charcoal and other environmental proxy techniques is inherently difficult in large, dynamic wetland systems due to variations in charcoal sources, sediment and charcoal deposition rates, and taphonomic processes. Major problems facing fire history reconstruction using macro-charcoal records in these wetlands include: (1) spatial and temporal variations in fire activity and ash and charcoal products within the wetlands, (2) variations in allochthonous inputs of charcoal from upstream sources, (3) tendency for geomorphic dynamism to affect flow dispersal and sediment and charcoal accumulation, and (4) propensity for post-depositional modification and/or destruction of macro-charcoal by flooding and taphonomic processes. Recognition of complex fire-climate-hydrology-vegetation interactions is essential. High-resolution, multifaceted approaches with reliable geochronologies are required to assess spatial and temporal patterns of fire and to reconstruct in order to interpret wetland fire regimes.

Wetland carbon storage controlled by millennial-scale variation in relative sea-level rise.

Coastal wetlands (mangrove, tidal marsh and seagrass) sustain the highest rates of carbon sequestration per unit area of all natural systems1,2, primarily because of their comparatively high productivity and preservation of organic carbon within sedimentary substrates3. Climate change and associated relative sea-level rise (RSLR) have been proposed to increase the rate of organic-carbon burial in coastal wetlands in the first half of the twenty-first century4, but these carbon-climate feedback effects have been modelled to diminish over time as wetlands are increasingly submerged and carbon stores become compromised by erosion4,5. Here we show that tidal marshes on coastlines that experienced rapid RSLR over the past few millennia (in the late Holocene, from about 4,200 years ago to the present) have on average 1.7 to 3.7 times higher soil carbon concentrations within 20 centimetres of the surface than those subject to a long period of sea-level stability. This disparity increases with depth, with soil carbon concentrations reduced by a factor of 4.9 to 9.1 at depths of 50 to 100 centimetres. We analyse the response of a wetland exposed to recent rapid RSLR following subsidence associated with pillar collapse in an underlying mine and demonstrate that the gain in carbon accumulation and elevation is proportional to the accommodation space (that is, the space available for mineral and organic material accumulation) created by RSLR. Our results suggest that coastal wetlands characteristic of tectonically stable coastlines have lower carbon storage owing to a lack of accommodation space and that carbon sequestration increases according to the vertical and lateral accommodation space6 created by RSLR. Such wetlands will provide long-term mitigating feedback effects that are relevant to global climate-carbon modelling.

Mangrove dynamics and blue carbon sequestration.

We monitored coastal wetland vertical accretion, elevation gain and surface carbon (C) at Homebush Bay, Australia over 18 years (2000-2017) in three settings initially characterized by saltmarsh, mixed saltmarsh-mangrove ecotone and mangrove-dominated zones. During this time, the saltmarsh transitioned to mixed saltmarsh-mangrove ecotone, and the mixed saltmarsh-mangrove ecotone transitioned to mangrove, consistent with vegetation transitions observed across the east Australian continent in recent decades. In spite of mangrove recruitment and thickening in the former saltmarsh zone, and the dominance of mangrove root material as a contributing C source, the rate of C accumulation in the former saltmarsh zone did not change over the study period, and there was no significant increase in surface elevation. This contrasted with the response of sites with a longer history of mangrove colonization, which showed strong accretion and C accumulation over the period. The result suggests that the C accumulation and surface elevation gains made as a result of mangrove colonization may not be observable over initial decades, but will be significant in the longer term as forests reach maturity.

New light in the dark - a proposed multidisciplinary framework for studying functional ecology of groundwater fauna.

Groundwaters provide the vast majority of unfrozen freshwater resources on the planet, but our knowledge of subsurface ecosystems is surprisingly limited. Stygofauna, or stygobionts -subterranean obligate aquatic animals - provide ecosystem services such as grazing biofilms and maintaining water quality, but we know little about how their ecosystems function. The cryptic nature of groundwaters, together with the high degree of local endemism and stygofaunal site-specific adaptations, represent major obstacles for the field. To overcome these challenges, and integrate biodiversity and ecosystem function, requires a holistic design drawing on classical ecology, taxonomy, molecular ecology and geochemistry. This study presents an approach based on the integration of existing concepts in groundwater ecology with three more novel scientific techniques: compound specific stable isotope analysis (CSIA) of amino acids, radiocarbon analysis (14C) and DNA analyses of environmental samples, stygofauna and gut contents. The combination of these techniques allows elucidation of aspects of ecosystem function that are often obscured in small invertebrates and cryptic systems. Carbon (δ13C) and nitrogen (δ15N) CSIA provides a linkage between biogeochemical patterns and ecological dynamics. It allows the identification of stygofaunal food web structures and energy flows based on the metabolic pathway of specific amino groups. Concurrently, 14C provides complementary data on the carbon recycling and incorporation within the stygobiotic trophic webs. Changes in groundwater environmental conditions (e.g. aquifer recharge), and subsequent community adaptations, can be pinpointed via the measurementof the radiocarbon fingerprint of water, sediment and specimens. DNA analyses are a rapidly expanding approach in ecology. eDNA is mainly employed as a biomonitoring tool, while metabarcoding of individuals and/or gut contents provides insight into diet regimes. In all cases, the application of the approaches in combination provides more powerful data than any one alone. By combining quantitative (CSIA and 14C) and qualitative (eDNA and DNA metabarcoding) approaches via Bayesian Mixing Models (BMM), linkages can be made between community composition, energy and nutrient sources in the system, and trophic function. This suggested multidisciplinary design will contribute to a more thorough comprehension of the biogeochemical and ecological patterns within these undervalued but essential ecosystems.

Itrax micro X-ray fluorescence (µXRF) for soft biological tissues.

Determination of the elemental composition of soft biological tissue is a time-consuming and tedious process when using traditional analytical techniques. In this method, micro X-ray fluorescence (µXRF) via Itrax, a scanning instrument, was used to determine elemental abundance at a resolution of 200 µm. Itrax µXRF was initially designed for elemental profiling of geological cores, and the capability of this technique was extended to soft biological tissue samples. The samples were dried and ground into a fine powder before analysis. The scanner generates elemental values as counts per 1 mm and these values are standardised to obtain the relative elemental abundance of the elements present in the samples. The acquired data can be used for environmental and biological research. •No literature could be found whereby the capability of Itrax µXRF has been extended to soft biological tissue samples.•The major advantages Itrax has over conventional methods is that it is a simultaneous technique which allows data to be acquired for over 30 elements at once with minimal sample preparation.•It is a non-destructive process where the samples can be re-used for additional analyses if necessary; this is especially useful when there is only a limited amount of sample available for other analyses.

Novel detection of provenance in the illegal wildlife trade using elemental data.

Despite being the fourth largest criminal market in the world, no forensic tools have been sufficiently developed to accurately determine the legal status of seized animals and their parts. Although legal trading is permissible for farmed or captive-bred animals, many animals are illegally removed from the wild and laundered by masquerading them as captive bred. Here we present high-resolution x-ray fluorescence (XRF) as a non-invasive and cost-effective tool for forensic classification. We tested the efficacy of this technique by using machine learning on a training set of zoo specimens and wild-caught individuals of short-beaked echidnas (Tachyglossus aculeatus), a small insectivorous monotreme in Australia. XRF outperformed stable isotope analysis (δ13C, δ15N), reducing overall classification error below 4%. XRF has the added advantage of providing samples every 200 µm on a single quill, enabling 100% classification accuracy by taking the consensus of votes per quill. This accurate and cost-effective forensic technique could provide a much needed in situ solution for combating the illegal laundering of wildlife, and conversely, assist with certification of legally bred animals.

Radiocarbon as a Novel Tracer of Extra-Antarctic Feeding in Southern Hemisphere Humpback Whales.

Bulk stable isotope analysis provides information regarding food web interactions, and has been applied to several cetacean species for the study of migration ecology. One limitation in bulk stable isotope analysis arises when a species, such as Southern hemisphere humpback whales, utilises geographically distinct food webs with differing isotopic baselines. Migrations to areas with different baselines can result in isotopic changes that mimic changes in feeding relations, leading to ambiguous food web interpretations. Here, we demonstrate the novel application of radiocarbon measurement for the resolution of such ambiguities. Radiocarbon was measured in baleen plates from humpback whales stranded in Australia between 2007 and 2013, and in skin samples collected in Australia and Antarctica from stranded and free-ranging animals. Radiocarbon measurements showed lower values for Southern Ocean feeding than for extra-Antarctic feeding in Australian waters. While the whales mostly relied on Antarctic-derived energy stores during their annual migration, there was some evidence of feeding within temperate zone waters in some individuals. This work, to our knowledge, provides the first definitive biochemical evidence for supplementary feeding by southern hemisphere humpback whales within temperate waters during migration. Further, the work contributes a powerful new tool (radiocarbon) for tracing source regions and geographical feeding.

Metal Transfer among Organs Following Short- and Long-Term Exposures Using Autoradiography: Cadmium Bioaccumulation by the Freshwater Prawn Macrobrachium australiense.

The uptake, depuration, and organ distribution of the radioisotope 109Cd were used to explore the internal kinetics of this nonessential metal following accumulation from waterborne cadmium by the freshwater decapod crustacean Macrobrachium australiense. Short- (6 h) and long-term (7 to 14 days) exposures to the radioisotope in solutions of 0.56 µg Cd/L were followed by depuration in metal- and isotope-free water for up to 21 days. The anatomical distribution of the radionuclide was visualized using autoradiography at predefined time points. The gills did not become saturated with cadmium after 14 days of exposure and demonstrated a greater rate of cadmium uptake relative to the hepatopancreas. Cadmium concentrations decreased rapidly during depuration from both gills and hepatopancreas after short exposures but slowly following long-term exposures. This suggests that the duration of cadmium exposure influences the depuration rate for this organism. The study demonstrates the complex behavior of cadmium accumulated by M. australiense and improves our understanding of how exposure duration will influence the internal location and potential toxicity of metals.

Inputs of anthropogenic nitrogen influence isotopic composition and trophic structure in SE Australian estuaries.

Urban development in coastal settings has increased the input of nitrogen into estuaries globally, in many cases changing the composition of estuarine ecosystems. By focussing on three adjacent estuaries with a gradient of anthropogenic N loadings, we used stable isotopes of N and C to test for changes due to increased anthropogenic N input on the structure of some key trophic linkages in estuaries. We found a consistent enrichment in δ(15)N corresponding to increased anthropogenic N at the three ecosystem levels studied: fine benthic organic matter, grazing invertebrate, and planktivorous fish. The degree of enrichment in δ(15)N between fine benthic organic matter and the grapsid crab Parasesarma erythrodactyla was identical across the three sites. The glassfish Ambassis jacksoniensis showed lower levels of enrichment compared to basal food sources at the higher N-loaded sites, suggesting a possible effect of anthropogenic N in decreasing food-chain length in these estuaries.

Bioaccumulation kinetics and organ distribution of cadmium and zinc in the freshwater decapod crustacean Macrobrachium australiense.

This study used the radioisotopes (109)Cd and (65)Zn to explore the uptake, retention and organ distribution of these nonessential and essential metals from solution by the freshwater decapod crustacean Macrobrachium australiense. Three treatments consisting of cadmium alone, zinc alone, and a mixture of cadmium and zinc were used to determine the differences in uptake and efflux rates of each metal individually and in the metal mixture over a three-week period, followed by depuration for 2 weeks in metal-free water using live-animal gamma-spectrometry. Following exposure, prawns were cryosectioned and the spatial distribution of radionuclides visualized using autoradiography. Metal uptake and efflux rates were the same in the individual and mixed-metal exposures, and efflux rates were close to zero. The majority of cadmium uptake was localized within the gills and hepatopancreas, while zinc accumulated in the antennal gland at concentrations orders of magnitude greater than in other organs. This suggested that M. australiense may process zinc much faster than cadmium by internally transporting the accumulated zinc to the antennal gland. The combination of uptake studies and autoradiography greatly increases our understanding of how metal transport kinetics and internal processing may influence the toxicity of essential and nonessential metals in the environment.