Age determination of orange roughy,Hoplostethus atlanticus (Pisces: Trachichthyidae) using210Pb:226Ra disequilibria

Natural levels of²¹⁰Pb:²²⁶Ra in otoliths of orange roughy,Hoplostethus atlanticus, from south-east Australian waters, were measured to determine fish ages radiometrically. Up to maturity, radiometric age estimates were consistent with a single constant otolith growth rate. Radiometric ages for juveniles were comparable with, but greater than, those obtained in a recent, validated New Zealand study which employed counts of annuli on the surface of otoliths. Beyond maturity, radiometric ages were obtained by modelling with an otolith growth rate set at 45% of the juvenile rate. Radiometric ageing confirms that orange roughy is very slow-growing, with an age at maturity (32 cm standard length, SL) of ~ 32 yr, and is very long-lived, with fish 38 to 40 cm being 77 to 149 yr old. These results have important implications for the management of the fishery.     

Stock structure of the southern bluefin tuna Thunnus maccoyii: an investigation based on probe microanalysis of otolith composition

Analysis of the chemistry of calcified tissues has been suggested to be a source of useful information on the population structure and environmental history of fishes. We have investigated this approach as a means of determining the number of spawning areas and diversity of migration routes in the large pelagic scombrid, Thunnus maccoyii (southern bluefin tuna). Analysis was based on ontogenetic variation in the composition of sagittal otoliths, as measured using two probe microanalysers (wavelength dispersive electron probe microanalysis and proton-induced X-ray emission microanalysis), of 9 larvae collected on the single known spawning ground (NE Indian Ocean), of 29 juveniles caught at different points along the known migration routes (off western Australia, southern Australia, and South Africa), and of 14 adults caught in the high-seas fishery (off SE Australia). Fifteen elements were detected in T. maccoyii sagittae, but only six (Ca, Na, Sr, K, S, and Cl) were consistently present at concentrations above minimum detection limits. No attempt was made to measure the concentrations of C, N and O, which are assumed to also be present. Comparisons among different samples indicated that: (1) variation in the composition of the otolith primordium was unimodal and, generally, normally distributed; (2) this composition varied among specimens as a function of their size or, apparently, year-class; (3) individuals collected from widely separated locations did not differ clearly in the composition of the most recently deposited sections of their otoliths; and (4) all variation in the composition of adult otoliths was encompassed in the range of variation of juveniles collected along the major known migration route. These observations are consistent with the hypothesis of a single spawning area for T. maccoyii, but also indicate that the range of environmentally correlated variation in composition is too low to provide a robust test of the diversity of migration routes. It is not clear why this variation is so low, but we suspect that it reflects both the relative homogeneity of the pelagic environment and a weak effect of environmental factors on the concentration of elements present in otoliths at levels1 ppm.     

Elemental composition of otolith growth marks in three geographically separated populations of European hake (Merluccius merluccius)

The composition of the opaque and translucent bands in the otoliths of European hake (Merluccius merluccius) from three different populations (Celtic Sea, Galicia shelf and Cretan Sea) was analysed with wavelength dispersive spectrometry (WDS) and differences in Ca, Sr and Na were compared. The translucent bands (annual marks and checks altogether) were significantly richer in Sr and Ca and significantly poorer in Na than opaque bands. Results support the idea of a more compact arrangement of crystals in translucent bands compared to opaque. Analyses did not help to discriminate between putative annual marks and checks. Clear differences in the range of Sr/Ca variation across the otolith as well as in the composition of the otolith core were observed among the three populations. As a result it was possible to discriminate the natal origin of each fish using the composition of the otolith core in Ca, Sr and Na. Nevertheless, the overall pattern across the otolith was similar among the three populations suggesting the existence of ontogenetic trends. Variation of Na/Ca ratios was largely uninformative. Based on these results, a conceptual model for hake is proposed which integrates the different sources of otolith Sr/Ca variation: ambient Sr/Ca, endogenous cues (ontogeny) and exogenous cues (environment).     

Chemistry of tuna otoliths: assessment of base composition and postmortem handling effects

Protocols used to collect and prepare otoliths for chemical analysis may result in either contamination or loss of elements, thus biasing population studies in unknown ways. We evaluated precision and bias associated with collection and cleaning procedures for three Atlantic tuna species: Atlantic bluefin tuna (Thunnus thynnus), yellowfin tuna (T. albacares), and blackfin tuna (T. atlanticus). Elemental concentrations were measured using solution-based inductively coupled plasma mass spectrometry and atomic absorption spectrophotometry. Seven elements were present above detection limits in all samples (Na, Mg, K, Ca, Mn, Sr, Ba). Mean concentrations of all seven elements were statistically indistinguishable in fresh pairs of otoliths of T. thynnus (mean error 5%, range 2-8%) and T. albacares (mean error 5%, range 3-7%); no indication of a left versus right otolith effect was observed. Otolith elemental concentrations were size dependent and significant inverse relationships were observed for Mg, Na, and K. Deliberate contamination of previously cleaned samples using a 10-ppm solution of a mixture of elements demonstrated that otoliths easily acquire surface contamination. Recleaning contaminated otoliths restored elemental concentrations to levels similar to control otoliths. Precision of paired comparisons between cleaned otoliths and those exposed to contamination and recleaned was high (mean error 6%). The effects of storage at two temperatures (7 days at -20°C, 3 days at 1°C) were investigated. For K, Ca, Sr, Mn, and Ba, variation between control (removed immediately) and treatment otoliths (in situ freezing or chilling) was similar to variation observed within fresh otolith pairs (mean error: fresh vs frozen 5%, fresh vs iced 5%). Statistically significant but small (<10%) postmortem storage effects were observed for Na and Mg. Estimates of error indexed to natural ranges in otolith chemistry of T. thynnus and T. albacares from different geographic regions in the Atlantic and Pacific showed that error values of several elements (Mg, Mn, Ba, Na, K) accounted for a small proportion of the natural range, suggesting levels of precision achieved in this study are suitable for the purpose of stock delineation.     

In situ experiment of ontogenetic variability in the otolith chemistry of Stegastes partitus

Otolith chemistry can be used to assess pelagic larval fish connectivity by comparing spatially variable otolith edge chemistry (corresponding to the site of collection) to otolith core chemistry (corresponding to the site of hatching). However, because the otolith’s edge and core represent different life stages, the deposition of elements may differ, thus complicating direct comparisons of edge and core chemistry to investigate connectivity. Here we present data from a field experiment in which otoliths from embryos (3 days post-fertilization) and juveniles of Stegastes partitus were collected at the same site and time, and chemically analyzed to assess whether elemental concentrations of otoliths vary ontogenetically. Separate multivariate analyses, each investigating the spatial/temporal variability in the chemistry of either embryo otoliths or the edges of juvenile otoliths, revealed significant differences, suggesting an environmental influence to the chemical signals of otoliths. A nested multivariate analysis assessing whether otolith chemistry varied with life history stage (i.e., ontogenetic variability) indicated that elemental concentrations of embryo otoliths were significantly greater than that of juvenile otolith edges. Specifically, embryo elemental concentrations of Mn, Zn, Sn, Ba, Ce, and Pb were between 2 and 163 times greater than those of the corresponding juvenile otoliths, and thus the environment was not the primary determinant of embryo otolith chemistry. Consequently, caution is warranted when interpreting environmental patterns of otolith cores, particularly when using them as a proxy for natal signatures.     

Population variation in the mitochondrial cytochrome b gene of the orange roughy Hoplostethus atlanticus and the hoki Macruronus novaezelandiae

To describe the genetic relationship among regional populations of two commercially valuable species of marine fish, the orange roughy Hoplostethus atlanticus and the hoki Macruronus novaezelandiae, the mitochondrial (mt) DNA haplotypes of 59 individuals were defined by direct sequencing of the cytochrome b gene. Samples of orange roughy were collected on four fishing grounds around New Zealand, one off the west coast of Tasmania and one near South Africa from 1990–1991. Samples from hoki were collected on three fishing grounds around New Zealand and one off Tasmania during the same period. An average of 252 nucleotides were sequenced from each of 32 orange roughy and an average of 372 nucleotides from each of 27 hoki. Sequence variation allowed the definition of 9 unique orange roughy haplotypes and 5 hoki haplotypes. Genetic variation, as measured by both average sequence divergence and haplotype diversity, was high in the orange roughy (nucleotide diversity=0.590%, haplotype diversity=0.782) and low in the hoki (nucleotide diversity=0.076%, haplotype diversity=0.279) relative to a similar survey of the Atlantic cod, Gadus morhua. Differences in haplotype frequencies of orange roughy from New Zealand, Tasmania and South Africa were not significant, and the most common haplotype was found in similar frequencies in these three geographically distant regions. Differences in haplotype frequencies between the New Zealand and Tasmanian samples of hoki were significant, suggesting restricted gene flow between these two regions. The contrasting patterns of low but regionally subdivided genetic variation in the hoki versus high but geographically undifferentiated genetic variation in the orange roughy may be attributed to the low fecundity, slow maturation and long lifespan of the orange roughy relative to the hoki.     

Effects of specimen handling and otolith preparation on concentration of elements in fish otoliths

The use of otolith chemistry as a tool for analysing the biology of fishes assumes that the procedures used to collect and prepare otoliths for analysis do not alter their composition. With otoliths of Nemadactylus macropterus, Hoplostethus atlanticus, and Rhombosolea tapirina, we show that this assumption is not valid for the elements that can be detected using electron-probe microanalysers (those present at concentrations greater than ≃100 parts per million): all six elements routinely measured using these techniques were affected by at least one post-mortem procedure tested. Measured concentrations of calcium and strontium were relatively insensitive to most procedures tested, whereas concentrations of sodium, potassium, sulphur and chlorine were affected substantially by many commonly used procedures. The ease with which otolith composition could be modified post-mortem suggests that apparent geographic, habitat-specific or ontogenetic differences in otolith composition should be interpreted with extreme caution because of easily induced artefacts and the problem of pseudo-replication associated with the ways otoliths are sampled and prepared for analysis. Received: 23 September 1996 / Accepted: 11 April 1998     

Estuarine dependency in a marine fish evaluated with otolith chemistry

Life-long strontium patterns in otoliths of the sciaenid Micropogonias furnieri caught in the southwestern Atlantic Ocean were examined to evaluate estuarine dependency and habitat use. Otolith Sr concentrations were on average 820?±?55?μg?g^?1 for freshwater, 1,751?±?101?μg?g^?1 for estuarine, and ranged from 2,000 to over 4,000?μg?g^?1 for marine waters. The examination of life-long otolith Sr revealed that 71?% of the marine-sampled fish moved toward brackish waters from age 0 to age 1, and that estuarine egress ranged from ages 2.1 to 4.1?years depending on the sampling area. Three different long-term patterns of Sr accumulation were observed and inferred to be the result of ontogeny and habitat shifts. Given that an estuarine Sr signature was consistently present in all sampled fish, M. furnieri is suggested to be a true estuarine-dependent species during its early life history.     

Elevated manganese concentrations at the cores of clupeid otoliths: possible environmental,physiological, or structural origins

Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was used to investigate the distribution of manganese (Mn) in Atlantic herring (Clupea harengus L.) otoliths. An unusual pattern of Mn concentration, which has not previously been reported in fish otoliths, was observed. Time-resolved plots (time roughly equivalent to ablation depth) of the Mn signal recorded in the ICP-MS detector during ablation of otoliths contained a distinct peak, suggesting that elevated Mn concentrations were specific to certain regions. Ablation inside and outside of the otolith core revealed that the region of high Mn concentration coincided with the ablation of the core. It was initially suggested that Mn from the gravel substrate on which herring eggs are incubated is incorporated into the otolith core, producing elevated Mn concentrations in this region. It was hypothesised that otoliths from fish whose eggs are not incubated in gravel will not contain high Mn concentrations at the core. To test this, otoliths of sprat (Sprattus sprattus), which have a pelagic embryological stage, and from reared herring, which were incubated on glass plates, were analysed using LA-ICP-MS. Peaks in the Mn signal were observed during the ablation of cores from some sprat and reared herring otoliths, indicating that incubation on a gravel substrate is not necessary for the accumulation of Mn at the otolith core and that otoliths from both pelagic and demersally spawning fish species can contain elevated Mn concentrations. The results suggest that Mn concentrations at the otolith core do not reflect the external environment of the developing embryo. Two other hypotheses are considered. A dramatic increase in Mn concentrations in the egg, associated with embryological development, may produce elevated Mn levels at the otolith core. Alternatively, the high levels of Mn observed at the otolith core may reflect spatial variation in the crystal structure of the calcium carbonate portion of the otolith. The possible influence of physiological or structural factors on the chemical composition of the otolith core may confound the environmental signal in this region of the otolith. This has important implications for the interpretation of otolith chemistry data.Communicated by J.P. Thorpe, Port Erin     

Exploring the potential of otolith microchemistry to enhance diet analysis in pinnipeds

Increasing the scope and accuracy of information about pinniped diets obtainable from non-invasive techniques is increasingly important, particularly in cases where pinniped species are threatened or endangered. This study is the first to explore the potential for using elemental analysis of the otoliths found in scat to enhance the information available for diet analyses. We investigated the effects of pinniped digestion on otolith microchemistry using laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS). We compared the elements contained in the edges (adult stage) and cores (larval/juvenile stage) of otoliths from Pacific herring (Clupea pallasii), Atka mackerel (Pleurogrammus monopterygius), and Walleye pollock (Theragra chalcogramma) recovered from the scat of captive Steller sea lions (Eumetopias jubatus) to elements in a sample of pristine (undigested) otoliths. We found that digestion had a significant effect on four of the six sampled combinations of species and otolith region (herring edges and cores, mackerel edges, and pollock cores), and that Rb most frequently showed significant differences in concentration after digestion. We could significantly discriminate among species of both pristine and digested otoliths using either otolith edges or cores with the elements Ba, Rb, Sr, Y, and Mg. When compared to previously identified digested otoliths, unknown samples of the three species of digested otoliths could be discriminated with 55–100% accuracy depending on species and otolith region. When compared to a library of previously identified pristine and digested otoliths, unknown samples of digested otoliths could be discriminated to species with 65–88% accuracy. When the group of unknown digested otoliths was compared to known pristine otoliths, discrimination ranged from 45 to 65%. These results indicate that elemental analysis could be used to supplement visual identification of otoliths from scat. However, further research is required to determine whether elemental analysis of digested otoliths could be useful for prey fish population studies.     

Scales of variation in otolith elemental chemistry of juvenile staghorn sculpin (<Emphasis Type="Italic">Leptocottus armatus</Emphasis>) in three Pacific Northwest estuaries

Although distinct otolith elemental signatures are often observed in fish collected from different estuaries, significant differences are also observed among sites within estuaries. Variation at these smaller spatial scales is not well quantified and has the potential to lead to inappropriate interpretations of otolith elemental data. To quantify variation at multiple scales, the otolith elemental composition (Mg:Ca, Mn:Ca, Sr:Ca, Ba:Ca, and Pb:Ca) of juvenile staghorn sculpin (Leptocottus armatus Girard, 1854) collected from five sites within three estuaries, the Columbia River (two sites) and Coos Bay (one site), Oregon, and Humboldt Bay, California (two sites), was examined. Using laser ablation-inductively coupled plasma mass spectrometry, each otolith was sampled at three zones: (1) within the primordium, which represents the egg and early larval periods; (2) at the outer edge, which represents the juvenile period just prior to collection; and (3) midway between the primordial and edge samples, which represents the late larval and early juvenile period. There were significant differences in otolith metal-to-calcium ratios at all scales examined. Using multi-element otolith signatures, fish were classified to estuary and site within estuary with relatively high levels of accuracy (av = 70–90%). The largest differences in metal-to-calcium ratios were observed between sites within estuaries (<5 km apart) and the smallest differences were observed among otolith zones. Variation in otolith chemistry may be used to provide information on probable habitat use by estuarine fish but studies must be carefully designed. Electronic supplementary material  Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Episodes of starvation are recorded in the otoliths of juvenile Ambassis vachelli (Chandidae), a tropical estuarine fish

Juvenile Ambassis vachelli, 40 to 50 d old, were used to test the effects of starvation and recovery-feeding on the widths of otolith increments. Three laboratory treatments (fed continuously; fed for 9d then starved for 15 d; starved for 9 d then fed for 15 d), were compared. Fish were also collected throughout the experiment from a field site to assess the natural fluctuations of increment widths. Growth, starvation and recovery were reflected in the widths of otolith increments. The width of increments deposited during starvation were similar in all fish, regardless of prior feeding history, despite the lower mortality recorded in starved fish with a history of intense feeding. Increment-width data were confounded by an ontogenetic decrease in increment widths with age. The results indicate that starvation and recovery are recorded in the increment widths of otoliths and are distinguishable from the effects of ontogenetic development. However, ontogenetic changes in the width of otolith increments confounded the interpretation of otolith-increment series. Therefore, the ontogenetic pattern of otolith increments must be known for each species before valid interpretation of otolith microstructure is possible.     

Isolation of Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>) nursery areas

This study examines the degree of isolation among 0-group Atlantic cod in a region where a number of resident adult congregations have been found. The location of key nursery areas in Scottish coastal waters was identified from trawl surveys of settled 0-group cod around the Scottish coast. Segregation among four of these key areas; within the Clyde, Shetland Isles, Moray Firth and Buchan was investigated using otolith elemental composition. Isotope concentrations in the otoliths of cod were analysed by solution-based inductively coupled plasma mass spectrometry of the whole otolith. Discriminant analyses showed a clear separation between nursery areas based on the concentrations of three elements (manganese, barium and magnesium). The concentration of these elements was not affected by the age or size of individuals. Classification accuracy exceeded 90% for all four sites. Evidence of temporal persistence in the relative spatial differences in elemental concentrations between two sites for 2 year-classes was found. The results suggest limited exchange between sites during some period of the early life history. The significance of this limited exchange is discussed in relation to recent evidence for adult fidelity and the utility of closed areas for protecting nursery grounds.     

Genetically isolated stocks of orange roughy (Hoplostethus atlanticus), but not of hoki (Macruronus novaezelandiae), in the Tasman Sea and Southwest Pacific Ocean around New Zealand

Restriction fragment-length polymorphisms of mitochondrial (mt) DNA, generated by six-base restriction enzymes, were used to examine the stock structure of the two major commercial species in New Zealand fisheries, orange roughy (Hoplostethus atlanticus Collett) and hoki (Macruronus novaezelandiae Hector). Samples of orange roughy from six spawning sites around New Zealand and one site off Tasmania show significant genetic heterogeneity. Samples from two sites around the south of New Zealand were genetically similar to each other, but dissimilar to samples from all other sites. The common haplotype observed in northern sites was absent from the two southern sites. Low numbers of unique haplotypes were restricted to the Challenger Plateau in the Tasman Sea. There was no significant genetic heterogeneity in five samples of hoki from two spawning sites off New Zealand and one site off Tasmania.     

Sclerochronological studies reveal that patterns of otolith growth of adults of two co-occurring species of Platycephalidae are synchronised by water temperature variations

Chronologies are developed from the otolith growth-increment widths of adult rock flathead (Platycephalus laevigatus) and longhead flathead (Leviprora inops) collected from three inter-connected embayments in temperate south-western Australia. Marginal increment trends on otoliths, in combination with the dendrochronological technique of crossdating, provide strong evidence that an opaque zone is formed annually in the otoliths of both species. Increment widths between opaque zones in the otoliths of individuals of P. laevigatus (maximum age 20 years) and L. inops (maximum age 21 years) were synchronised within and between species. The correlation between the two master otolith chronologies, i.e. mean standardised increment width for each year, was highly significant (P < 0.01). The master otolith chronology for each species was positively correlated with mean sea surface temperatures from 1 July to 30 June, which encompassed the austral summer when growth is typically greatest, and with mean monthly sea surface temperature between September and March. Leeuwin Current strength, which influences the biotic characteristics of some marine species on the south coast, has little influence on the pattern of otolith growth, presumably because the inter-connected embayments from which fish were obtained were partially enclosed. On the basis of the above data and the geographical distributions of P. laevigatus and L. inops, it is proposed that, while both species would grow faster with increasing temperature, the temperatures on the south coast are closer to the optimum for the former species.     

Low levels of global genetic differentiation and population expansion in the deep-sea teleost <Emphasis Type="Italic">Hoplostethus atlanticus</Emphasis> revealed by mitochondrial DNA sequences

The orange roughy Hoplostethus atlanticus is a well-known commercial species with a global distribution. There is no consensus about levels of connectivity among populations despite a range of techniques having been applied. We used cytochrome c oxidase subunit I (COI) and cytochrome b sequences to study genetic connectivity at a global scale. Pairwise Φ_ST analyses revealed a lack of significant differentiation among samples from New Zealand, Australia, Namibia, and Chile. However, low but significant differentiation (Φ_ST = 0.02–0.13, P < 0.05) was found between two Northeast Atlantic sites and all the other sites with COI. AMOVA and the haplotype genealogy confirmed these results. The prevalent lack of genetic differentiation is probably due to active adult dispersal under the stepping-stone model. Demographic analyses suggested the occurrence of two expansion events during the Pleistocene period.     

Changes in otolith strontium:calcium ratios in metamorphosing Conger myrisaster leptocephali

Content ratios of strontium (Sr) to calcium (Ca) in the otolith of Conger myriaster metamorphosing leptocephali and elvers increased with increasing increment number from the core to the 110th increment and subsequently decreased. The otolith region from the 110th increment to the edge corresponded to the metamorphic stage. The Sr:Ca ratios in otolith edges of metamorphosing leptocephali were inversely related to metamorphic stage, suggesting that the changes in otolith Sr:Ca ratios were influenced by some physiological factor(s) rather than by environmental factors. Sr concentration in leptocephalus somatic tissues was high and decreased as metamorphosis progressed until the late metamorphic stage when the preanal myomere to total myomere ratio was 0.4. Ca concentration was constant throughout ontogenesis. Body Sr:Ca ratios markedly decreased as metamorphosis progressed. Decrease in somatic Sr concentration and the consequent decrease in body Sr:Ca ratios seemed to be associated with the breakdown of glycosaminoglycan (GAG) in gelatinous matrix, which is the major constituent of soft tissue in leptocephali. Catabolism of GAG may also cause a decrease in otolith Sr:Ca ratios during metamorphosis. In leptocephalus otoliths, Sr:Ca ratios may change in association with the synthesis and breakdown of GAGs during ontogeny. Received: 29 November 1996 / Accepted: 6 January 1997     

Climatic effects on the growth of a temperate reef fish from the Southern Hemisphere: a biochronological approach

Increments in the hard parts of marine organisms (otoliths, skeletons, shells) can provide long-term chronologies of growth analogous to tree rings. For the first time in the Southern Hemisphere, we use a dendrochronological (tree-ring analysis) approach to develop a multidecadal chronology of growth for a temperate reef fish, Girella tricuspidata, from the coast of northern New Zealand. Growth patterns in the otoliths of this species were strongly synchronous among individual fish over a period spanning 27 years (1980–2006). We then compared our otolith chronology to climatic records and found strong positive correlations of growth with sea surface temperature, and weak negative correlations with the multivariate El Nino Southern Oscillation (ENSO) index. Strongest correlations were found between summer sea surface temperature and otolith growth. This relationship was consistent across all years and explained 44 % of the variation (y = −2.0 + 0.1785 × temperature, r ² = 0.4367, P = 0.0002) in the G. tricuspidata growth chronology. Our study illustrates how otolith chronologies provide remarkable records of annual growth patterns over decadal time scales that will be useful for forecasting the likely effects of climate change on marine ecosystems.