The effect of otolith storage methods on the concentrations of elements detected by laser-ablation ICPMS

Otoliths of male (1+ year old) Terubok, tropical shad Tenualosa toli from the Lupar River, Sarawak, were treated in different ways to assess the effect of post-capture storage on element concentrations in the otolith core and edge detected by laser-ablation inductively coupled plasma mass spectrometry (ICPMS). Five treatments were compared: rapid removal and dry storage; fish kept 5 h after capture then otoliths removed; rapid removal and stored in alcohol; fish rozen for 24 h then otoliths removed; and storage of intact head in alcohol for several weeks. Seven elements were detected at the core and edge of otoliths (Li, Na, Mg, Mn, Co, Sr, Ba). Of these, Na, Mg, Co and Ba differed among treatments, being highest in treatments where the otolith remained in the head. Variation in element measurements due to treatment effects was similar to within-otolith variation for the other elements and was least variable in the fresh treatment. The variation in concentration between the core and edge was more significant than other sources of variation (up to 100 times) for some elements. These differences were probably due to physiological factors (ontogenetic) or to yearly changes in water chemistry. The results indicate that the method of otolith storage, especially freezing whole fish, can have a small, but measurable, effect on the concentrations of Na, Mg, Co and Ba.     

Otolith elemental fingerprints of juvenile Pacific swordfish Xiphias gladius

The trace element composition of young‐of‐the‐year (YOY) juvenile swordfish Xiphias gladius sagittal otoliths were analysed as a preliminary test of the value of otolith elemental fingerprints for determining swordfish nursery ground origins in the central Pacific Ocean. A suite of five elements (Mg, Zn, Sr, Ba and Pb) was assayed with isotope dilution ICP‐MS; all elemental concentrations were roughly comparable to otoliths of other marine fishes. Multivariate analyses of elemental fingerprints based on Ba and Sr revealed differences between sample sites, and the magnitude of the differences increased with latitudinal separation. With more comprehensive sampling of nursery grounds, it should be possible to identify origin of nursery ground for adult swordfish by analysing the YOY juvenile portion of the sagittal otolith.     

The detection of elements in larval otoliths from Atlantic herring using laser ablation ICP-MS

Trace element concentrations of otoliths from larval herring Clupea harengus collected from known spawning beds in the Celtic and Irish Seas, were investigated using laser ablation ICP-MS and compared with concentrations in the larval cores of juvenile otoliths from the same populations and year class. A range of elements (Mg, Zn, Sr, Ba and Pb) was detectable in early larval otoliths (20–40 µm diameter). Larval otolith concentrations exceeded the larval core concentrations of juvenile otoliths and also the concentrations reported in the literature, for Mg, Zn, Ba and Pb, indicating that the measurement of elements in larval otoliths was severely affected by post-mortem contamination, most likely due to adherence of tissue and endolymph residue on the otolith surface. Comparison of otolith composition between larvae from two freezing treatments showed that contamination from Mg and Zn was more serious in otoliths that had remained in frozen larvae for prolonged periods. Larval populations from the two seas showed significant differences in otolith Sr concentrations, which were consistent over two sampling years. Similar differences were seen in the corresponding juvenile populations. The results show that while early larval otoliths are extremely susceptible post-mortem contamination, Sr concentrations can be reliably measured using laser ablation ICP-MS and for this element, the detection of region specific differences is possible.     

The differentiation of Stegastes partitus populations using lapillar and sagittal otolith chemistry

The comparison of elemental concentrations of sagittal and lapillar otoliths from the same individuals of Stegastes partitus indicated significant differences for several elements. Sagittal otoliths were superior at differentiating individuals, yet the differentiation of individuals was further improved when the elemental concentrations of both otolith types were used in the same analysis.     

Mn<Superscript>2+</Superscript> concentrations in coastal fish otoliths: understanding environmental and biological influences from EPR

The Mn²⁺ concentrations in the sagittae otoliths of 12 fish families (and 19 species) that co-occur in a coastal area of southeastern Brazil (~21°S) were quantified using electron paramagnetic resonance (EPR). Inferences were made about the relationship between fish habitat and trace element incorporation. Inferences were made on the relationship between trace element concentration and otolith shape. The differences in Mn²⁺ concentrations among the species suggest that habitat (and feeding habits) might drive the incorporation of this trace element into fish otoliths, with higher values in bottom-associated fish species than in surface-associated species. In surface-associated fish species, the correlation between trace element concentrations and otolith shape was stronger than in bottom-associated species. Thus, while the Mn bioavailability in a fish’s habitat, especially from feeding resources, is a local driving influence of trace element incorporation in sagittae otoliths, species-specific requirements also have an influence. Quantitative EPR is a non-destructive technique that is very useful when the available samples cannot be damaged, like with otolith collections.     

Elemental composition of otoliths from Atlantic croaker along an estuarine pollution gradient

Elemental compositions of otoliths of young-of-the-year Atlantic croaker Micropogonias undulatus at stations along a pollution gradient in Galveston Bay, Texas, were compared with those of sediments and livers to establish the level of consonance between otolith and habitat chemistries. Both coupled and uncoupled trends were indicated, with the bulk of the results pointing to the need for additional research to test the working hypothesis that the otolith stores a temporal record of elemental chemistry of resident waters. Lack of simple correlations between elemental concentrations in sediment or liver and in otoliths indicate that other biological and geochemical factors influence otolith composition.     

Spatial variability of trace elements in fish otoliths: comparison with dietary items and habitat constituents in seagrass meadows

Juvenile trumpeters Pelates sexlineatus , two main prey items (amphipods and polychaetes) and seagrass constituents (detritus, young and old leaves of Zostera capricorni ) were collected from two sites within six estuaries, to compare the spatial variability of elemental composition (Li, Mn, Sr, Cd, Ba and Pb). Average elemental values were lower in the fish otoliths except for Sr. Significant differences in elemental composition of otoliths were detected among estuaries (Mn and Pb). Spatial differences in concentrations of Cd were found in amphipods and polychaetes; the latter also showed differences for Ba and Pb. Detritus and Z. capricorni leaves showed appreciable spatial differences for most of the elements. Spatial variability between sites was generally high. There were differences in the multivariate composition of trace elements found in the otoliths of fish among estuaries. One estuary was consistently separated from the other estuaries in the ordinations. Significant correlations were detected between concentrations of Mn in otoliths and concentrations in prey, Z. capricorni and detritus. A significant relationship was also detected between Pb in otoliths and detritus. There were significant correlations between concentrations of Mn, Sr and Ba in detritus and that in otoliths. The study demonstrated that trophic transfer and habitat constituents may be considered as a potential source for some of the elemental accumulation in fish otoliths.     

Can otolith elemental chemistry retrospectively track migrations in fully marine fishes?

Otolith microchemistry can provide valuable information about stock structure and mixing patterns when the magnitude of environmental differences among areas is greater than the cumulative influence of any vital effects. Here, the current understanding of the underlying mechanisms governing element incorporation into the otolith is reviewed. Hard and soft acid and base (HSAB) theory is employed to explore the differences in chemical behaviours, distributions and affinities between elements. Hard acid cations (e.g. Mg(2+) , Li(+) and Ba(2+) ) tend to be less physiologically influenced and accepted more readily into the otolith crystal lattice but are relatively homogeneous in seawater. Soft acid cations (e.g. Zn(2+) and Cu(2+) ) on the other hand, exhibit more varied distributions in seawater, but are more likely to be bound to blood proteins and less available for uptake into the otolith. The factors influencing the geographical distribution of elements in the sea, and their incorporation into the otoliths of marine fishes are reviewed. Particular emphasis is placed on examining physiological processes, including gonad development, on the uptake of elements commonly used in population studies, notably Sr. Finally, case studies are presented that either directly or indirectly compare population structuring or movements inferred by otolith elemental fingerprints with the patterns indicated by additional, alternative proxies. The main obstacle currently limiting the application of otolith elemental microchemistry to infer movements of marine fishes appears to lie in the largely homogeneous distribution of those elements most reliably measured in the otolith. Evolving technologies will improve the discriminatory power of otolith chemistry by allowing measurement of spatially explicit, low level elements; however, for the time being, the combination of otolith minor and trace element fingerprints with alternative proxies and stable isotopic ratios can greatly extend the scope of migration studies. Among the otolith elements that routinely occur above instrument detection limits, Ba, Mn and Li were deemed the most likely to prove reliable geographic markers in marine species.     

Small-scale spatial variation in the elemental composition of otoliths of Stegastes nigricans (Pomacentridae) in French Polynesia

Solution-based inductively coupled plasma-mass spectrometry (ICP-MS) was used to determine if Stegastes nigricans collected from 15 sites in French Polynesia could be distinguished by the trace element composition of their otoliths. A total of 293 adults were collected by spearing and their otoliths were analysed. We found that elemental signatures differed significantly among sites within and between the islands of Tahiti and Moorea (p<0.001), primarily due to variation in concentrations of the elements Ba, Ca, Li, Mg, Mn, Na, Sr and Y. The otoliths of fish collected within Papeete Harbour in Tahiti had distinctive elemental signatures characterised by relatively high concentrations of Mn. Otoliths of these fish could be distinguished from others that were collected only a small distance (200 m) from the harbour. This is the first time that differences in chemical composition of otoliths have been reported at such small spatial scales and this trait may prove useful for the studies of connectivity of populations at within reef scales.     

Analysis of the soluble matrix of vaterite otoliths of juvenile herring (Clupea harengus): do crystalline otoliths have less protein?

Otoliths are calcium carbonate concretions laid down in the inner ear of fish and used in fish age estimation. Otoliths precipitate in the form of aragonite but aberrant precipitation may result in vaterite formation instead of aragonite. Vaterite otoliths are more translucent than aragonite. The quantity of HCl-soluble proteins (SP) was measured in the vaterite otoliths and their aragonite pairs of one year old reared herring Clupea harengus to assess the changes induced by the precipitation of vaterite in the amount of soluble proteins in the otolith. Results showed that vaterite otoliths had as much soluble proteins as their aragonite pairs (p>0.05). Due to the lower density of the vaterite, vaterite otoliths were lighter than their aragonite pairs (p<0.05) which explained that protein concentrations were significantly higher (p<0.05) than in aragonite otoliths. These results indicate that the precipitation of vaterite in otoliths did not affect the inclusion of soluble proteins. Furthermore, they suggest that soluble proteins do not affect the translucent or opaque appearance of otoliths. Differences in translucency may instead be caused by the amounts of insoluble proteins or by differences in the physical properties of proteins. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of the otolith proteins revealed two bands at 50 and 62 kDa in both aragonite and vaterite otoliths suggesting that the precipitation of vaterite in the otolith is not controlled by either of these two proteins present in the otolith.     

Otolith growth and age estimation in the European hake

The internal sulcal rings in Merluccius merluccius otoliths cannot be considered as annuli. In the absence of a strong seasonal Zeitgeber, hake otoliths in the Mediterranean did not lay down an interpretable ring pattern that would be useful for age determination. A total of 484 sagittal otoliths from specimens ranging between 6 and 94 cm L T was studied in monthly samples from the Gulf of Lions in 1989–1990. Transverse, burnt otolith sections was analysed with an image analysis system using enhanced and filtered images and using a fast Fourier transform (FFT) function to avoid subjectivity in ring interpretation. The otolith radius-fish length relationship showed allometric growth and sexual dimorphism. The ring pattern was consistent for the sulcal rings in both sexes. Changes in the marginal increment showed the formation of multiple sulcal rings, of both environmental and physiological origin. The ring pattern depended on the sex and sexual activity.     

An evaluation of the otolith characteristics of Conger conger during metamorphosis

A sample of 20 metamorphosing conger eel Conger conger leptocephali were collected from the Minho River, Portugal, in February 1999 and their sagittal otoliths were analysed by scanning electron microscopy. Four different etching agents were applied along both sagittal and frontal sections during otolith preparation to examine the microstructural growth in this species. Otolith growth increments were visible throughout the increment countable zone using all four treatments, but a permanent peripheral diffuse zone, where the daily increments were unclear, appeared on all otoliths, preventing accurate age estimation. To understand more about the nature of the diffuse zone, otoliths of 10 other metamorphosing leptocephali reared in aquaria were marked by immersion in tetracycline hydrochloride. The distance between the fluorescent marks and otolith edge, measured over a fixed period of time, was used to estimate the otolith growth rate. The application of this technique led to an anomalously high estimated otolith growth rate, probably as a result of the capture, marking and handling stress.     

Analysis of otolith chemistry in Nassau grouper (Epinephelus striatus) from the Bahamas and Belize using solution-based ICP-MS

 We examined the utility of otolith minor and trace element chemistry, assayed with inductively coupled plasma mass spectrometry (ICP-MS), as a means of delineating population structure in the Nassau grouper (Epinephelus striatus). We characterized the elemental composition of otoliths collected in 1993 from three locations in Exuma Sound, Bahamas and from Glover Reef, Belize in 1995. A single location in Exuma Sound was sampled in 1994 to test temporal variability in otolith composition. Five elements (Ca, Zn, Sr, Ba and Pb) were routinely detected, at levels significantly above background, by solution-based ICP-MS. Results from analysis of variance of elemental data, expressed as a ratio to Ca, indicated that there were no significant differences among the Exuma locations for any element, but significant variability was found between Glover Reef and the pooled Exuma localities for Zn/Ca, Sr/Ca and Ba/Ca ratios. Significant inter-annual differences at one Exuma Sound location was restricted to Ba/Ca ratios. Discriminant function analysis correctly classified 86% and 95% of the Belize and pooled Exuma sites, respectively. Otoliths from Belize were characterized by low Zn/Ca and high Ba/Ca and Pb/Ca ratios compared to otoliths from fish collected in Exuma Sound. Although differences in Ba levels may be related to upwelling at Glover Reef, more data are needed to definitely link otolith composition with regional differences in water chemistry. Accepted: 15 February 1999     

Trace elements in the otoliths and dorsal spines of albacore tuna (Thunnus alalunga, Bonnaterre, 1788): An assessment of the effectiveness of cleaning procedures at removing postmortem contamination

Trace element analysis or “elemental fingerprinting” is widely used in stock structure analyses. Postmortem contamination of bony structures can confound the results of microconstituent studies or introduce an additional source of noise to the data, thus reducing the ability of the technique to detect real variation in trace element concentrations. Despite the potential for postmortem contamination during sample preparation, the effectiveness of the procedures used to remove potential contaminants from sectioned otoliths and other calcareous structures prior to laser ablation inductively coupled plasma mass spectrometry (LA ICP-MS) has not previously been addressed. Otoliths and dorsal spine sections of albacore tuna (Thunnus alalunga) collected from the North East Atlantic Ocean and the Mediterranean Sea were deliberately contaminated prior to analysis of trace element composition using LA ICP-MS. The effectiveness of three cleaning treatments (rinsing in ultrapure water, 30% hydrogen peroxide and ultrapure 5% nitric acid) at removing this postmortem contamination were compared. Magnesium and strontium were relatively robust to postmortem effects when exposed to contamination at concentrations of 50 ppm and 200 ppm respectively. Soaking in a solution containing Mn, Cs and Ba (50 ppm) caused a marked increase in the detected concentration of each element in both structures. Translucent bands in both structures were more susceptible to contamination. Rinsing in ultrapure water or hydrogen peroxide was not effective at removing Mn, Cs and Ba contamination from either calcareous structure. Washing the otoliths and spines in nitric acid successfully removed postmortem contaminants.The removal of otoliths from tuna damages the appearance of the fish and has an adverse effect on market value. However spines are easily removed, do not affect the appearance or value of the fish and are the most commonly used structure for age determination. A weak but significant correlation was observed between Ba in opaque zones in otoliths and dorsal spines. All other spine to otolith correlations were not significant. The results do not provide support for the use of spines as an alternative to otoliths in trace elemental analyses.     

Contribution of water chemistry and fish condition to otolith chemistry: comparisons across salinity environments

This study quantified the per cent contribution of water chemistry to otolith chemistry using enriched stable isotopes of strontium (⁸⁶Sr) and barium (¹³⁷Ba). Euryhaline barramundi Lates calcarifer, were reared in marine (salinity 40), estuarine (salinity 20) and freshwater (salinity 0) under different temperature treatments. To calculate the contribution of water to Sr and Ba in otoliths, enriched isotopes in the tank water and otoliths were quantified and fitted to isotope mixing models. Fulton's K and RNA:DNA were also measured to explore the influence of fish condition on sources of element uptake. Water was the predominant source of otolith Sr (between 65 and 99%) and Ba (between 64 and 89%) in all treatments, but contributions varied with temperature (for Ba), or interactively with temperature and salinity (for Sr). Fish condition indices were affected independently by the experimental rearing conditions, as RNA:DNA differed significantly among salinity treatments and Fulton's K was significantly different between temperature treatments. Regression analyses did not detect relations between fish condition and per cent contribution values. General linear models indicated that contributions from water chemistry to otolith chemistry were primarily influenced by temperature and secondly by fish condition, with a relatively minor influence of salinity. These results further the understanding of factors that affect otolith element uptake, highlighting the necessity to consider the influence of environment and fish condition when interpreting otolith element data to reconstruct the environmental histories of fish.     

Otolith chemistry of Champsodon nudivittis (Ogilby, 1895) and Nemipterus randalli (Russell, 1986) in Iskenderun Bay,Turkey

The study objective was to investigate the chemical composition of otoliths of two Lessepsian fish migrants, namely, Champsodon nudivittis and Nemipterus randalli, which thrive in the Iskenderun Bay, Turkey. The study specifically investigated the age structure and explored differences in chemical otolith composition in relation to age. Samples were collected using a traditional Mediterranean bottom trawl (mesh size 44 mm) at depths of 45 to 90 m. A total of 78 Champsodon nudivittis (size range, 6.0 to 14.0 cm) and 60 Nemipterus randalli (size range, 6.1 to 17 cm) were captured in May 2012. Age readings were carried out (sectioning technique). Additionally, the concentrations of Na, K, Li, and Ca were determined using flame photospectrometry. The results revealed that the concentrations of Na (5.70 mg/g) and K (4.45 mg/g) in otoliths of Nemipterus randalli were predominant elements after Ca (128.71 mg/g). The concentration of Li in otoliths was also statistically different in the two species. This study contributes to the knowledge of the otolith chemistry in the two Lessepsian fish species now living in the same (but new) geographical region.     

Elemental signatures of Acanthochromis polyacanthus otoliths from the Great Barrier Reef have significant temporal, spatial, and between-brood variation

We evaluated the spatial and temporal scales over which otolith signatures varied in a reef fish on the Great Barrier Reef (GBR) using the non-dispersing damselfish Acanthochromis polyacanthus. We found a robust multi-element separation in otolith signatures from reef clusters in the northern and southern GBR. Variance components indicated that this spatial scale accounted for the majority of the variation in two elemental ratios (Ba/Ca and Sr/Ca) over the 2 years of the study. There was also significant variation in elemental signatures between otoliths collected over two consecutive years, as well as within a season. Individual reefs within clusters were less distinguishable based on otolith chemistry and were probably observed by differences within reefs (among sites and broods within sites). These results indicate that it may be difficult to determine the reef of origin for individual fish using otolith chemistry, while determining natal region seems a realistic goal.     

Effect of diet on otolith composition in Pomatomus saltatrix, an estuarine piscivore

To test the hypothesis that elemental composition of otoliths (sagittae) could be influenced by differences in natural prey type, young‐of‐the‐year bluefish Pomatomus saltatrix were captured immediately after their migration from oceanic waters into mid‐Atlantic Bight estuaries and fed either shrimp, Crangon septemspinosa and Palaemonetes spp. or fish Menidia menidia under similar temperature and salinity regimes in two separate 60 day experiments. Unlimited rations of fish and shrimp prey were provided in the first experiment which led to differences in bluefish growth rate between the two prey treatments; fish prey was limited in the second experiment to ensure that growth rates of bluefish in the two prey treatments were similar. Concentrations of seven elements in bluefish otoliths were determined using solution‐based inductively coupled plasma mass spectrometry (ICPMS). There was no significant effect of diet on five of the seven elements examined (Na, Mg, K, Ca and Mn). The levels of Sr and Ba in the otoliths of shrimp‐fed bluefish, however, were significantly higher than fish‐fed bluefish in both experiments. Concentrations of Ba in shrimp‐fed bluefish otoliths were double that found in fish‐fed bluefish. The results suggest that diet can explain some of the variation in otolith chemistry previously attributed to physical and chemical properties of the water.     

Comparison of spatial variation in otolith chemistry of two fish species and relationships with water chemistry and otolith growth

Spatial variation in the chemistry (Mg, Mn, Sr and Ba) of recently deposited otolith material (last 20–30 days of life) was compared between two demersal fish species; snapper Pagrus auratus (Sparidae) and sand flathead Platycephalus bassensis (Platycephalidae), that were collected simultaneously at 12 sites across three bays in Victoria, south-eastern Australia. Otolith chemistry was also compared with ambient water chemistry and among three sampling positions adjacent to the proximal otolith margin. For both species, variation in otolith chemistry among bays was significant for Ba, Mn and Sr; however, differences among bays were only similar between species for Ba and Mn. Only Ba showed significant variation at the site level. Across the 12 sites, mean otolith Ba levels were significantly positively correlated between species. Further, although incorporation rates differed, mean ambient Ba levels for both species were positively correlated with ambient Ba levels. Spatial variation in multi-element otolith chemistry was also broadly similar between species and with multi-element water chemistry. Partition coefficients clearly indicated species-specific incorporation of elements into otoliths. Mg and Mn were consistently higher in snapper than sand flathead otoliths (mean ±s .d ., Mg snapper 22·1 ± 3·8 and sand flathead 9·9 ± 1·5 μg g⁻¹, Mn snapper 4·4 ± 2·6 and sand flathead 0·5 ± 0·3 μg g⁻¹), Sr was generally higher in sand flathead otoliths (sand flathead 1570 ± 235 and snapper 1346 ± 104 μg g⁻¹) and Ba was generally higher in snapper otoliths (snapper 12·1 ± 12·8 and sand flathead 1·8 ± 1·4 μg g⁻¹). For both species, Mg and Mn were higher in the faster accreting regions of the otolith margin, Sr was lower in the slower accreting region and Ba showed negligible variation among the three sampling regions. This pattern was consistent with the higher Mg and Mn, and generally lower Sr observed in the faster accreting snapper otoliths. It is hypothesized that the differences between species in the incorporation of these elements may be at least partly related to differences in metabolic and otolith accretion rate. Although rates of elemental incorporation into otoliths appear species specific, for elements such as Ba where incorporation appears consistently related to ambient concentrations, spatial variation in otolith chemistry should show similarity among co-occurring species.     

基于椭圆傅里叶分析的次南极电灯鱼矢耳石形态多样性

次南极电灯鱼矢耳石形态特征具有多样性.为了深入研究其形态特征,利用南设得兰群岛外侧水域采集的456尾次南极电灯鱼(体长范围6.0～8.8 cm)样本,对其矢耳石形态进行分析和判别.根据形态特征将次南极电灯鱼耳石分为4种类型,并采用椭圆傅里叶分析法选取表征耳石类型的77个傅里叶特征系数进行了分析.结果表明: 对4种耳石类型两两比较后发现,具有显著性差异的傅里叶特征系数最多及最少分别占总体的61%和28.6%;对77个傅里叶系数进行主成分分析,前22个主成分解释了总变异的76.6%;选取了17个傅里叶特征系数进行判别分析,建立判别函数,总体判别率为87.2%;根据椭圆傅里叶分析重建的耳石轮廓反映了4种耳石类型间的差异.4种耳石类型在不同体长及体质量的次南极电灯鱼中皆有出现,表明耳石类型具有随机性,且左右耳石类型不一致,表明其左右耳石外形具有差异性.4种耳石类型中,Ⅰ型和Ⅱ型占总体的72.6%,为次南极电灯鱼耳石的主要形态;Ⅲ型和Ⅳ型占总体的27.4%,为次要形态.