基于多测点LA-ICPMS的耳石核区元素指纹分析技术及其在鱼类群体识别中的实证研究

构建了多测点激光剥蚀取样（LA）-ICPMS元素分析-剥蚀点显微结构验证的耳石核区元素指纹分析技术,以此进行了五个刀鲚群体（辽东湾、渤海湾、黄河口、胶州湾及长江口群体）识别的实证研究。结果发现,耳石核区Sr：Ca和Ba：Ca比值是有效识别各地理群体的元素指纹,基于二者的各群体判别成功率在46.2%—92.3%之间,总体...     

渤、黄海大泷六线鱼耳石微化学特征与群体判别

为探究中国近海大泷六线鱼(Hexagrammosotakii)耳石微化学成分特征及群体分化状况,采集了连云港、青岛、威海、烟台、秦皇岛、大连、丹东7个野生群体和1个威海养殖群体的大泷六线鱼样本。利用LA-ICPMS技术测定耳石中的微量元素,以微量元素与钙的比值进行了分析,结果显示大泷六线鱼耳石中, Na/Ca、Sr/Ca、Fe/Ca、Mg/Ca较高,而Mn/Ca、Ba/Ca、Zn/Ca、Li/Ca较低。耳石1龄前元素比值的组间比较(S-N-K检验)结果显示:Na/Ca、Sr/Ca、Fe/Ca在群体间具显著性差异(P<0.05)。耳石边缘区元素与环境中元素比较发现,Li、Mn元素在耳石中富集程度较大,Sr在耳石的中元素含量远远大于水环境中的浓度,说明耳石中的元素沉积与海水中元素并不是简单的线性关系。判别分析结果显示:耳石1龄前元素指纹的判别分析结果表明,大泷六线鱼群体的判别成功率为63.0%,判别正确范围为30.0%～80.0%,耳石边缘区元素的判别正确率为57.1%,判别正确范围20.0%～80.0%,秦皇岛群体有高的判别成功率80.0%。耳石元素指纹可以用于不同大泷六线鱼群体的...     

Evidence for discrete subpopulations of sea perch (Helicolenus ercoides) across four fjords in Fiordland,New Zealand

In coastal populations of invertebrates and fishes, the distribution of discrete subpopulations is influenced by adult and larval dispersal, as well as by the effects of habitat heterogeneity on site fidelity or connectivity. Here, we examine evidence for spatial structure of sea perch, Helicolenus percoides, populations among four fjords in the Fiordland region of southwestern New Zealand. We examine patterns in adult morphology, length-at-age, δ¹³C and δ¹⁵N of muscle tissue, and trace elemental composition of whole otoliths as proxies for population isolation among the four inner fjord regions. A multivariate analysis of morphometrics reveals significant differences among populations from each of the four sites, suggesting existence of four distinct subpopulations. These patterns are consistent with observed differences in δ¹³C and δ¹⁵N, and length-at-age estimates among the four subpopulations. Differences in whole otolith concentrations of Sr, Ba, Mg and Li, and high classification scores based on the whole otolith elemental fingerprint are also consistent with significant subdivision among areas. Patterns across all four markers are consistent with discrete subpopulation structure of adult sea perch among the four study sites. These data indicate that the newly implemented network of marine protected areas in Fiordland is likely to contain discrete populations of sea perch.     

Oxygen isotopic distribution along the otolith growth axis by secondary ion mass spectrometry: Applications for studying ontogenetic change in the depth inhabited by deep-sea fishes

This study using tuna otoliths as working standards established a high lateral resolution and precision analysis to measure δ¹⁸O_otolith by secondary ion mass spectrometry. This analytical approach of the ion probe was applied to deep-sea fishes to reconstruct the likely depths inhabited by the fishes at different life history stages based on the measured δ¹⁸O_otolith values as a proxy of water temperature. Dramatic increases up to 5–6‰ in δ¹⁸O_otolith, representing a temperature decrease of approximately 20 °C, were detected in a blind cusk eel (Barathronus maculatus) otolith and in the otoliths of Synaphobranchus kaupii during leptocephalus metamorphosis to glass eel, inferred from the drop of otolith Sr/Ca ratios and increase of otolith growth increment width. δ¹⁸O_otolith profiles clearly divided the fish's life history into a planktonic stage in the mixed layer of the ocean and a benthic stage on the deep-sea ocean bottom. The habitat shift signal was recorded within a 150 µm width of otolith growth zone, which was too narrow to be clearly detected by mechanical drilling and conventional isotopic ratio mass spectrometry. However, variations down to −7‰ were found in δ¹⁸O_otolith profiles as the result of Cs²⁺ beam sputter in the core and larval portions of the otoliths. Carbon mapping by electron probe microanalyzer and staining by toluidine blue suggested abundant proteins existed in the areas with anomaly negative δ¹⁸O_otolith values, which cannot be interpreted as a habitat change but due to the isotopic fractionation by O emission from the proteins. These results implied that careful design and understanding of the chemical composition of the analytical areas or tracks on the heterogeneous otolith was essential for highly accurate and precise analysis.     

我国瓯江口和珠江口凤鲚洄游模式和生境利用研究

Habitat use of the tapertail anchovy(Coilia mystus Linnaeus, 1758) from the Oujiang River Estuary and the Zhujiang(Pearl) River Estuary was studied by examining the environmental signatures of Sr and Ca in otoliths using electron probe microanalysis. Individuals from the Oujiang River had higher and varied Sr:Ca ratios(expressed as(Sr:Ca)×1 000, 3.83–13.0 average) in the otolith core regions, suggesting that they were born in brackish or sea waters, and that a freshwater habitat might not be necessary for egg hatching and larval growth.While, individuals from the Zhujiang River had lower Sr:Ca ratios(0.39–2.51 average) in the core regions,suggesting a freshwater origin. After hatching, anchovies from the Zhujiang River migrate downstream to the river estuary close to brackish water. Our results demonstrated varied habitat use for spawning during stages of early life history between the two populations, and suggested that such variations are promoting diversity of life history strategies of this species.     

Spatio-temporal and interspecific variation in otolith trace-elemental fingerprints in a temperate estuarine fish assemblage

We tested whether estuarine fishes have site-specific differences in the concentrations of trace elements in their otoliths that can be used as ‘fingerprints’ to identify them to their estuary of origin. To evaluate the robustness of this approach, we tested whether elemental fingerprints were consistent among individuals of five species that were collected in 1996 from three temperate estuaries in southern California. We also tested whether elemental fingerprints were consistent between spring and autumn 1996 for three species in one of the sites, Carpinteria Marsh. The species evaluated comprised a mid-water-dwelling smelt (Atherinops affinis), two benthic gobies (Clevelandia ios and Ilypnus gilberti), and two flatfish (Paralichthys californicus and Hypsopsetta guttulata). The concentrations of six elements (Mn, Cu, Zn, Sr, Ba, and Pb) were determined in the otoliths using inductively coupled plasma-mass spectrometry (ICP-MS). Within estuaries, the five species exhibited strong variation in elemental concentration, indicating substantial interspecific differences in otolith environmental history. When the five fish species were considered separately, multivariate (MANOVA) and univariate (ANOVA) analyses of variance indicated that the elemental composition of otoliths differed significantly among the estuaries in four of the five species. Based on linear discriminant function analyses (DFA), differences were strong enough that trace element composition could be used to accurately assign fish to their site of origin [mean (range): 93.5% (74–100%)]. However, elemental signatures within Carpinteria Marsh were not consistent between spring and autumn 1996, and this was reflected in a substantial reduction in the accuracy of assigning fish to their true site of origin. When we compared site differences between fish species (site×species interactions), the elemental fingerprints were most similar between closely related species (e.g. the two gobies and the two flatfish) and most dissimilar between distantly related species, both phylogenetically and ecologically. Among the six elements analyzed, Sr and Ba exhibited the most inconsistent pattern among species, with significant differences in 80 and 70% of the pairwise species comparisons, respectively. The remaining four elements showed ≥70% consistency in the pattern of variation among sites for the different species. Thus, while otolith elemental fingerprinting can be a useful tool for inferring estuarine residency, such fingerprints may be temporally variable and species specific.     

Stock Identification of Chum Salmon (<Emphasis Type="Italic">Oncorhynchus keta</Emphasis>) Using Trace Elements in Otoliths

The relationship between trace elements in chum salmon otoliths and in their rearing water was investigated to develop ways to distinguish chum salmon stocks in Korea. Rearing water and otoliths of hatchery-reared chum salmon fry were collected from three major hatcheries (Yangyang, Samchuk, and Uljin) on the east coast of Korea in spring 2001 and 2002. Trace elements in the otoliths and rearing water were analyzed using inductively coupled plasma mass spectrometry (ICP-MS) at the Korea Basic Science Institute. The chemical composition of rearing water and otoliths of the salmon fry at specific sites did not vary significantly through the study period. The ratios of some trace elements to Ca in rearing water, such as Sr/Ca and Ba/Ca, was clearly reflected in the chemical composition of otoliths, although the absolute concentrations were different. These two elements may be useful in distinguishing between salmon hatcheries, which have different chemical environments. Discriminant analysis with the ratios of eight elements to Ca (Na/Ca, Mg/Ca, Al/Ca, Cu/Ca, Sr/Ca, Ba/ Ca, Mn/Ca, and Zn/Ca) revealed a distinct separation of natal area in stocks. The analysis of otolith chemistry may be an effective technique for identifying the origins of wild salmon caught at sea.     

黄海南部野生银鲳和鮸鱼的耳石元素微化学研究

利用电子显微探针元素分析技术(EPMA)对黄海南部野生银鲳和鮸鱼的耳石进行了锶和钙沉积特征的初步分析。定量线分析结果表明,两种鱼类耳石的Sr/Ca之间存在显著的种间差异。银鲳耳石Sr/Ca比,在耳石核心及相邻处为低值区(5.86±0.92);3段Sr/Ca高值区分别为近核心部(7.88±1.28)、第1龄处(9.44±1.82)及耳石边缘(7.91±1.38);揭示银鲳孵化和早期发育应需要盐度适中的生境,当龄鱼在后期的生长中需洄游经过两段高盐生境(其中之一在第1龄时)。鮸鱼耳石Sr/Ca比波动表现为耳石核心处(7.72±0.97)高于其余部分,反映了鮸鱼孵化及初期发育阶段可能生活在高盐度生境,而当龄鱼随后阶段的生长和发育过程则会洄游至盐度有所降低的生境中进行。     

Connectivity clues from short-term variability in settlement and geochemical tags of mytilid mussels

The use of geochemical tags in calcified structures of fish and invertebrates is an exciting tool for investigating larval population connectivity. Tag evaluation over relatively short intervals (weeks) may detect environmental and ecological variability at a temporal scale highly relevant to larval transport and settlement. We collected newly settled mussels (Mytilus californianus and M. galloprovincialis) weekly during winter/spring of 2002 along the coast of San Diego, CA, USA, at sites on the exposed coast (SIO) and in a protected coastal bay (HI), to investigate temporal patterns of geochemical tags in mussel shells. Analyses of post-settlement shell via LA-ICP-MS revealed statistically significant temporal variability for all elements we examined (Mg, Mn, Cu, Sr, Cd, Ba, Pb and U). Despite this, our ability to distinguish multielemental signatures between sites was largely conserved. Throughout our 13-week study, SIO and HI mussels could be chemically distinguished from one another in 78–87% of all cases. Settlement varied between 2 and 27 settlers gram-byssus^?1 week^?1 at SIO and HI, and both sites were characterized by 2–3 weeks with “high” settlement. Geochemical tags recorded in early larval shell of newly settled mussels differed between “high” and “low” settlement weeks at both sites (MANOVA), driven by Mg and Sr at SIO (p = 0.013) and Sr, Cd, Ba and Pb at HI (p < 0.001). These data imply that shifts in larval sources or transport corridors were responsible for observed settlement variation, rather than increased larval production. In particular, increased settlement at HI was observed concurrent with the appearance of geochemical tags (e.g., elevated Cd), suggesting that those larvae were retained in upwelled water near the mouth of the bay. Such shifts may reflect short-term changes in connectivity among sites due to altered transport corridors, and influence the demography of local populations.     

Habitat use by the European eel Anguilla anguilla in Irish waters

The apparent use of marine and freshwater habitats by European eel Anguilla anguilla was examined by analyzing the strontium (Sr) and calcium (Ca) concentrations in otoliths of the eels collected from Irish coastal and fresh waters. The age and growth of eels were also examined using their otolith annuli. The sizes and ages of the female eels were greater than those of the males. The somatic growth rates ranged from 15 to 62 mm/year, which is typical for Ireland and other European countries. Analyses of Sr:Ca ratios along a life history transect in each otolith showed peaks (maximum more than 25 × 10⁻³) between the core and elver mark corresponding to the period of their leptocephalus and early glass eel stages in the ocean. Outside the elver mark, the Sr:Ca ratios indicated that eels had remained in different habitats that included freshwater (average Sr:Ca ratios, 0.98–1.78 × 10⁻³) and areas with relatively high salinities (average Sr:Ca ratios, 6.73–8.89 × 10⁻³). Some individuals showed clear evidence of shifts from sea to fresh waters. These findings suggest that Irish eels have the same behavioral plasticity regarding whether or not to enter freshwater or remain in marine environments as has been recently documented in this species and several other temperate anguillid species. However, patterns of habitat use in Irish waters were somewhat different than those previously reported for other habitats.     

长江口水域四种鱼类的耳石微化学研究

利用X-射线电子探针微区技术(electron probe microanalysis, EPMA),对长江口不同水域中捕获的焦氏舌鳎、皮氏叫姑鱼、鮸鱼和光泽黄颡鱼耳石中的Sr含量进行耳石微化学研究,结果发现,焦氏舌鳎是典型的海水鱼类,不仅Sr:Ca比(按惯例标准化为Sr:Ca×10~3)的移动平均值高(7),而且Sr含量面分析图亦呈现为对应高盐度海水的黄色或红色图谱。虽然皮氏叫姑鱼和鮸鱼从出生直至被捕Sr:Ca比的移动平均值都在3—7间波动,总体上属河口半咸水栖息鱼类,但从Sr含量面分析图来看,皮氏叫姑鱼生活史履历更为复杂,可包括淡水(对应蓝色图谱)和半咸水生境(对应绿色图谱);而鮸鱼仅在长江口较高盐度半咸水或海水生境中活动,未见进入过淡水的履历。光泽黄颡鱼表现出仅利用淡水生境的履历,其Sr:Ca比的移动平均值仅在1.5—3间窄幅波动,整个生活史均在淡水区域里活动,表现为长江口典型的淡水栖息鱼类。本研究从新的角度提供了较为客观、直观和最新的信息,用以较为准确地重建和掌握长江口这些不同鱼类的生境利用特征。     

基于耳石中锶和钙元素分析破解日本六角川和筑后川河口水域刀鲚的生境履历

为了解日本有明海沿岸六角川和筑后川河口水域鳀科鲚属鱼类刀鲚（Coilia nasus）的洄游特征和生境履历,利用电子探针微区分析（EPMA）技术,对采于两水域刀鲚的矢耳石进行了锶（Sr）元素含量的X射线强度面分布及耳石最长轴锶钙元素比值（Sr:Ca）的定量线分析。结果发现10尾六角川刀鲚（LJC）中的7尾以及15尾筑后川刀鲚（ZHC）耳石核心区域的Sr:Ca比值≤ 3,显示出这些个体在淡水生境出生并早期生活于淡水的履历。紧邻核心区域随后的生长阶段,Sr面分布图谱显示出了多样对应于含量水平差异的不同颜色同心环轮变化样式,Sr:Ca比值的波动亦可相应地分成1-6个差异极其显著的阶段,显示出这些个体在随后生活史阶段中极富变化的生境转换状况。另外3尾六角川刀鲚耳石核心的Sr:Ca比值较高（3-6.7）,对应于半咸水生境;但在核心区域之外,Sr:Ca比值则高低变化不一,显示出这些六角川刀鲚个体在河口半咸水生境孵化的特征。本研究表明,二维Sr元素含量面分布图谱和基于耳石最长轴的Sr:Ca比值完全可以作为重建日本有明海沿岸各河口水域刀鲚生境履历的有效工具。     

Otolith microchemical evidence revealing multiple spawning site origination of the anadromous tapertail anchovy (Coilia nasus) in the Changjiang (Yangtze) River Estuary

The estuarine tapertail anchovy(Coilia nasus) is a high-value commercial fish. Estimating the spawning site or hatchery origin and habitat is essential for its conservation. This study aimed to determine the habitat use and life history characteristics of C. nasus from the Changjiang River Estuary. We investigated the environmental signatures of strontium(Sr) and calcium(Ca) in the otoliths of the collected specimens using electron probe microanalysis; additionally, we examined their gonadal mat...     

Classification of juvenile flatfishes to estuarine and coastal habitats based on the elemental composition of otoliths

The elemental composition of otoliths from juvenile English sole Pleuronectes vetulus and speckled sanddab Citharichthys stigmaeus living in estuaries and sandy coastal habitats were compared to determine if a chemical habitat tag existed that could be used to differentiate fish living in the alternative juvenile habitats. Juveniles of both species were collected from seven estuaries and 11 coastal sites along 500 km of the central California coast. Collections occurred in three years: 1998, 1999 and 2000. The chemical composition of the otoliths was analyzed using solution-based inductively coupled plasma mass spectrometry. The multi-elemental composition (Sr, Li, Ba, Mn) of otoliths from estuarine and coastal fish differed significantly for all models (global, region and years models) examined in this study. For each species, discriminant function analysis (DFA) for the global model, which pooled juveniles collected from three regions over multiple years, classified fish into estuarine and coastal groups with close to 80% accuracy. The two main elements in the discriminant models, Sr and Li, differed consistently between habitats; Sr was higher and Li was lower in estuarine fish. Classification success was modestly improved by generating separate discriminant functions for each region (regions model) because of some regional differences in Sr and Li concentrations. Classification success also was improved in some cases by generating separate discriminant functions for each year (years model) because Ba and Mn differed between habitats in only some years. Despite this variability in the concentration of elements in some regions and years, a chemical habitat tag was present in each species that could be used to discriminate coastal and estuarine juveniles over a large geographic area and over three years with very different oceanographic conditions (e.g., El Niño, La Niña). In addition, I found that English sole and speckled sanddab had striking similarities in their chemical habitat tags and that, in some cases, one species could be used as a proxy to classify juveniles of the other species without compromising the accuracy of the habitat tag. The ability to use a proxy classification model would significantly reduce the number of juvenile fish that would need to be collected and analyzed in order to classify members of an ecologically similar species. The chemical habitat tags found in this study appears to be promising tools for identifying fish that have lived in alternative juvenile habitats and, ultimately, the proportional contribution of estuarine and coastal habitats to the central California populations of English sole and speckled sanddab.     

Spatial variation in elemental composition of otoliths of three species of fish (family Sparidae)

Determining the nursery habitat of fishes that have moved from estuarine nursery habitats is difficult. The elemental fingerprints of otoliths of three species of sparids were determined to investigate their utility as a natural tag of the nursery habitat. Juvenile Pagrus auratus (snapper), Rhabdosargus sarba (tarwhine) and Acanthopagrus australis (bream) were collected from two sites in each of 15, six and three estuaries, respectively, and their otoliths analysed by solution-based inductively coupled plasma-mass spectrometry. Significant differences in otolith chemistry were found for all three species of juveniles collected from different estuaries. The same patterns among estuaries were not seen for all species, although it was not possible to sample the same sites within an estuary for all species. For bream, significant differences in otolith chemistry were found among all three estuaries, whereas for tarwhine the six estuaries were separated into three groups. For snapper, a number of estuaries could be separated, but there was some overlap for other estuaries. All three species were collected from the same site within one estuary and their otoliths analysed. Significant differences were found among species, but the implication of this finding remains unclear as the three species show differences in microhabitat use and may also differ in age. Because the elemental fingerprints of juveniles vary among estuaries or groups of estuaries, the nursery or recruitment estuary of adult fish could now be determined by analysing the juvenile region of adult otoliths. Thus, connectivity between estuaries and open coastal populations could be determined. Such information will have major implications for fisheries management because it will provide information on the distance that fish have moved from their recruitment estuary and the number of estuaries that contribute to each adult population.     

Stock identification and life history interpretation using trace element signatures in salmon otoliths

Chum salmon (Oncorhynchus keta) in the North Pacific Ocean are anadromous fish, and spend most of their life in the sea until spawning in natal streams. To identify the stock and habitat characteristics of chum salmon, the composition of chemical elements (Ca, Mn, Sr, Zn, and Ba) in otolith was examined using laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS). Two main types of analytical work have been carried out; discrete spot analysis and line scan analysis of otolith sections. Salmon otoliths were obtained from the eastern (Canada and USA) and western (Japan and Korea) North Pacific during 1997–1999 spawning seasons. Spot analysis of otolith cores demonstrated significant differences in the element concentration among countries (p = 0.003). Line scanning from the core to the margin showed that Sr concentrations were elevated at the core of the otoliths, decreased during the freshwater stage, increased suddenly at a certain point, and oscillated periodically towards the margin matching with year-ring. The elevated Sr concentration at the core may reflect the maternal contribution to the egg, and the oscillations toward the margin may reflect salinity gradients between onshore/offshore or north/south migrations. The Zn profiles also oscillated and corresponded to the annual ring of the otolith. However, the profiles of Sr and Zn oscillated oppositely after salmon migrated to saline water and the Zn uptake declined toward the rim of the otolith while Sr uptake increased.     

Barium variation in Pagrus auratus (Sparidae) otoliths: A potential indicator of migration between an embayment and ocean waters in south-eastern Australia

Chronological variation in otolith chemistry can be used to reconstruct migration histories of fish. The use of otolith chemistry to study migration, however, requires knowledge of relationships between the chemical properties of the water and elemental incorporation into otoliths, and how water chemistry varies in space and time. We explored the potential for otolith chemistry of snapper, Pagrus auratus, to provide information on movement history between a large semi-enclosed bay, Port Phillip, and coastal waters in south-eastern Australia. Comparisons of water chemistry across two years demonstrated that ambient barium (Ba) levels in Port Phillip Bay were approximately double those in coastal waters (11 μg L⁻¹ versus 6 μg L⁻¹). Ba levels in otolith margins of wild juvenile snapper were highly positively correlated with ambient levels across 17 sampling locations, and levels in otolith margins of adult snapper collected from Port Phillip Bay were approximately double those of snapper collected in coastal waters. Mean partition coefficients for Ba (D_Ba) were similar for juvenile (0.43) and adult (0.46) otoliths, suggesting that otolith Ba incorporation relative to ambient levels was similar across life-stages. Low Ba variation across otoliths from adult snapper maintained in tanks for three years indicated that annual temperature and/or growth cycles did not strongly influence otolith Ba variation. We concluded that chronological Ba variation in snapper otoliths would be a reliable proxy for life-history exposure to variable ambient Ba. We used water chemistry data and Ba levels across otoliths of ocean resident snapper to estimate otolith Ba levels indicative of residence in Port Phillip Bay (>10 μg g⁻¹) or coastal waters (<6 μg g⁻¹). Peaks in Ba exceeding 10 μg g⁻¹ were common across otoliths of snapper collected in Port Phillip Bay and a nearby coastal region. The location of strong Ba peaks within otoliths was consistent with residence in Port Phillip Bay during the spring/summer when snapper move into the Bay from coastal waters to spawn. Our results for snapper support the use of otolith Ba as a proxy for ambient levels throughout the life-history, however, confident interpretation of migration history from otolith Ba chronologies will most likely require matching time series of ambient Ba in the water bodies of interest.     

Application of laser ablation ICPMS to trace the environmental history of chum salmon Oncorhynchus keta

Trace element levels in otoliths of chum salmon Oncorhynchus keta were examined by means of laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS). A close linear relationship in the Sr:Ca ratios between EPMA (X-ray analysis with an electron microprobe) and LA-ICPMS analyses was found (p<0.0001), suggesting that the latter technique could be used to separate the marine and freshwater life phases. Mg:Ca, Cr:Ca, Zn:Ca and Ba:Ca ratios in either the core region or the oceanic growth zone of the otoliths varied among sites. These differences suggest that elemental compositions may reflect environmental variability among spawning (breeding) or habitat sites. Thus, those element ratios demonstrate the potential to be used to distinguish between fish spawning (breeding) sites and habitats for this species of salmon.     

The effect of habitat and environmental history on otolith chemistry of barramundi Lates calcarifer in estuarine populations of a regulated tropical river

We examine the microchemistry of otoliths of cohorts of a fished population of the large catadromous fish, barramundi Lates calcarifer from the estuary of a large tropical river. Barramundi from the estuary of the large, heavily regulated Fitzroy River, north-eastern Australia were analysed by making transects of ⁸⁷Sr/⁸⁶Sr isotope and trace metal/Ca ratios from the core to the outer edge. Firstly, we examined the Sr/Ca, Ba/Ca, Mg/Ca and Mn/Ca and ⁸⁷Sr/⁸⁶Sr isotope ratios in otoliths of barramundi tagged in either freshwater or estuarine habitats that were caught by the commercial fishery in the estuary. We used ⁸⁷Sr/⁸⁶Sr isotope ratios to identify periods of freshwater residency and assess whether trace metal/Ca ratios varied between habitats. Only Sr/Ca consistently varied between known periods of estuarine or freshwater residency. The relationships between trace metal/Ca and river flow, salinity, temperature were examined in fish tagged and recaptured in the estuary. We found weak and inconsistent patterns in relationships between these variables in the majority of fish. These results suggest that both individual movement history within the estuary and the scale of environmental monitoring were reducing our ability to detect any patterns. Finally, we examined fish in the estuary from two dominant age cohorts (4 and 7 year old) before and after a large flood in 2003 to ascertain if the flood had enabled fish from freshwater habitats to migrate to the estuary. There was no difference in the proportion of fish in the estuary that had accessed freshwater after the flood. Instead, we found that larger individuals with of each age cohort were more likely to have spent a period in freshwater. This highlights the need to maintain freshwater flows in rivers. About half the fish examined had accessed freshwater habitats before capture. Of these, all had spent at least their first two months in marine salinity waters before entering freshwater and some did not enter freshwater until four years of age. This contrasts with the results of several previous studies in other parts of the range that found that access to freshwater swamps by larval barramundi was important for enhanced population productivity and recruitment.     

Evidence of estuarine nursery origin of five coastal fish species along the Portuguese coast through otolith elemental fingerprints

Connectivity is a critical property of marine populations, particularly for species with segregated juvenile and adult habitats. Knowledge of this link is fundamental in understanding population structure and dynamics. Young adults of commercially important fish species Solea solea, Solea senegalensis, Platichthys flesus, Diplodus vulgaris and Dicentrarchus labrax were sampled off the Portuguese coast in order to establish preliminary evidence of estuarine nursery origins through otolith elemental fingerprints. Concentrations of Li, Na, Mg, K, Mn, Cu, Zn, Sr, Ba and Pb in the otolith section corresponding to juvenile's nursery life period were determined through laser ablation inductively coupled plasma mass spectrometry. Element: Ca ratios in coastal fish differed significantly amongst collection areas, except for Platichthys flesus, and were compared with the elemental fingerprints previously defined for age 0 juveniles in the main estuarine nurseries of the Portuguese coast. Identification of nursery estuaries was achieved for four of the species. Assigned nursery origins varied amongst species and differences in the spatial scale of fish dispersal were also found. Diplodus vulgaris was not reliably assigned to any of the defined nurseries. Overall, results give evidence of the applicability of estuarine habitat tags in future assessments of estuarine nursery role. Research developments on the links between juvenile and adult habitats should contribute for the integrated management and conservation of nurseries and coastal stocks.