The use of otolith chemistry to characterize diadromous migrations

Chemical constituents in otoliths have become a valuable tool for fish ecologists seeking to reconstruct migratory patterns and life-history diversity in a wide range of species worldwide. This approach has proved particularly effective with fishes that move across substantial salinity gradients over the course of their life, including many diadromous species. Freshwater endmembers of several elemental and isotope ratios (e.g. Sr:Ca, Ba:Ca and (87)Sr:(86)Sr) are typically identifiably distinct from marine values, and often differ among freshwater tributaries at fine spatial scales. Because these chemical tags are generally incorporated in proportion to their ambient dissolved concentrations, they can be effective proxies for quantifying the presence, duration and frequency of movements between freshwater and marine habitats. The development of high precision probe-based analytical techniques, such as laser ablation inductively coupled plasma mass spectrometry (ICP-MS) and microbeam methods, has allowed researchers to glean increasingly detailed life-history profiles of these proxies across otoliths. Researchers are also combining multiple chemical proxies in an attempt to refine interpretations of habitat residence patterns. A thorough understanding of the spatial and temporal variation in water chemistry as well as environmental and physiological controls on incorporation of specific elements into otoliths is required for confident estimation of lifetime salinity experience. First some assumptions, methodological considerations and data processing options that are particularly relevant to diadromous otolith chemistry studies are discussed. Insights into diadromous migrations obtained from decades of otolith chemistry research, highlighting the increasingly recognized importance of contingent behaviour and partial migration are then discussed. Finally, areas for future research and the need to integrate otolith chemistry studies into comprehensive assessments of the effects of global environmental change are identified.     

Incorporation of strontium into otoliths of an estuarine fish

Patterns of Sr/Ca variability in fish otoliths have been widely applied as tracers of movement between freshwater and marine habitats, with the assumption that low salinity habitats correspond to lower otolith levels of Sr/Ca. On the other hand, fluvial estuaries can contain steep gradients in Sr/Ca, and in some estuaries, freshwater values of Sr/Ca can exceed marine values, which are relatively constant across marine habitats. Therefore, to interpret Sr/Ca variability in otoliths of fish that move through estuaries, information is needed about both the incorporation of strontium into otoliths and the nature of the gradient of Sr/Ca in the water. We conducted four experiments to evaluate the incorporation of strontium into fish otoliths under estuarine conditions, using white perch (Morone americana) as a model estuarine fish. One laboratory and the two field experiments tested the relationship between Sr/Ca in the otolith and that in the water. A fourth experiment investigated the effect of salinity, independently of the water chemistry (Sr was manipulated while maintaining a constant salinity and Ca level). All four experiments supported a direct relationship between Sr/Ca in the otolith and the water, across a range of estuarine salinities. Results also indicated that the incorporation of strontium into otoliths of estuarine fishes should be constant across broad gradients of Sr/Ca in estuarine waters. While the experiments supported past applications of tracing estuarine and diadromous movements with otolith Sr/Ca chronologies, we emphasize the need to understand the underlying nature of Sr/Ca gradients in estuaries, which may limit or confound reconstructions of estuarine habitat use.     

Use of otolith chemistry to examine patterns of diadromy in the threatened Australian grayling Prototroctes maraena

Otolith chemical analyses (proton-induced X-ray emission and laser ablation inductively coupled plasma mass spectrometry) were used to examine patterns of diadromy in Australian grayling Prototroctes maraena collected from three rivers in south-eastern Australia. Concentrations of Sr:Ca and Ba:Ca were measured in the sagittal otoliths of 25 fish and in water samples collected on two occasions from multiple sites within freshwater, estuarine and marine reaches of the rivers. The results provide evidence of marine residency during the juvenile phase, with high Sr:Ca and low Ba:Ca in the inner region of the otoliths that were consistent across all three rivers. This general pattern was apparent for all individuals examined, suggesting that diadromous migration may be an obligate aspect of the life history. The chemical signatures of the outer regions of the otoliths were distinct between the three rivers, whilst there was no difference in the inner regions of the otoliths. This suggests that juveniles from different rivers may reside in a relatively homogenous chemical environment, such as the sea, and populations in coastal Victorian rivers may share a common marine recruitment source.     

Spatial and temporal variations in otolith chemistry and relationships with water chemistry: a useful tool to distinguish Atlantic salmon Salmo salar parr from different natal streams

Otolith elemental (Sr:Ca, Ba:Ca, Mn:Ca, Mg:Ca and Rb:Ca) and isotopic (⁸⁷Sr:⁸⁶Sr) profiles from several annual cohorts of juvenile Atlantic salmon Salmo salar were related to the physico‐chemical characteristics (chemical signatures, flow rate, temperature and conductivity) of their natal rivers over an annual hydrological cycle. Only Sr:Ca, Ba:Ca and ⁸⁷Sr:⁸⁶Sr in otoliths were determined by their respective ratios in the ambient water. Sr:Ca ratios in stream waters fluctuated strongly on a seasonal basis, but these fluctuations, mainly driven by water flow regimes, were not recorded in the otoliths. Otolith Sr:Ca ratios remained constant during freshwater residency at a given site and were exclusively related to water Sr:Ca ratios during low flow periods. While interannual differences in otolith elemental composition among rivers were observed, this variability was minor compared to geographic variability and did not limit classification of juveniles to their natal stream. Success in discriminating fish from different sites was greatest using Sr isotopes as it remained relatively constant across years at a given location.     

Low interbasin connectivity in a facultatively diadromous fish: evidence from genetics and otolith chemistry

Southern smelts (Retropinna spp.) in coastal rivers of Australia are facultatively diadromous, with populations potentially containing individuals with diadromous or wholly freshwater life histories. The presence of diadromous individuals is expected to reduce genetic structuring between river basins due to larval dispersal via the sea. We use otolith chemistry to distinguish between diadromous and nondiadromous life histories and population genetics to examine interbasin connectivity resulting from diadromy. Otolith strontium isotope (⁸⁷Sr:⁸⁶Sr) transects identified three main life history patterns: amphidromy, freshwater residency and estuarine/marine residency. Despite the potential for interbasin connectivity via larval mixing in the marine environment, we found unprecedented levels of genetic structure for an amphidromous species. Strong hierarchical structure along putative taxonomic boundaries was detected, along with highly structured populations within groups using microsatellites (F_ST = 0.046–0.181), and mtDNA (Φ_ST = 0.498–0.816). The presence of strong genetic subdivision, despite the fact that many individuals reside in saline water during their early life history, appears incongruous. However, analysis of multielemental signatures in the otolith cores of diadromous fish revealed strong discrimination between river basins, suggesting that diadromous fish spend their early lives within chemically distinct estuaries rather than the more homogenous marine environment, thus avoiding dispersal and maintaining genetic structure.     

基于耳石微化学的长江安徽和县江段刀鲚生境履历重建

为了解长江刀鲚(Coilia nasus)的“生境履历”, 利用电子探针微区分析技术研究了2015年5月17日在长江安徽和县江段捕获的刀鲚短颌鲚与长颌鲚类型耳石Sr和Ca微化学特征。短颌鲚的耳石Sr/Ca值变化动态为2类, 一类比值稳定在2.00左右(1.65±0.87—2.03±0.96), 反映了其纯淡水的生境履历; 另一类比值波动显著, 不仅具有对应淡水生境的低值(2.39±1.18—2.63±0.85), 还具有对应半咸水生境的高值(3.22±1.17—3.29±1.14), 显现了其江海生活的生境履历。长颌鲚耳石的Sr/Ca值均波动显著, 同时具有对应淡水生境的低值(1.35±1.07—2.37±1.04)和半咸水、海水生境的高值(3.47±1.09—8.35±1.25), 表现了其均为溯河洄游的生境履历。Sr面分析结果也与上述Sr/Ca值的结果相印证。长江安徽和县江段刀鲚资源组成复杂, 群体可由纯淡水型和江海生活型的个体共同组成。     

Freshwater habitat use by a moray eel species,Gymnothorax polyuranodon,in Fiji shown by otolith microchemistry

Freshwater eels of the Anguillidae are diadromous because they migrate between ocean and freshwater environments, but other anguilliform fishes are generally considered to be strictly marine species. A few marine eels of the Muraenidae and Ophichthidae have occasionally been found in freshwater or estuaries, indicating that anguillids are not the only anguilliform eels that can use freshwater in some parts of the world. The moray eel Gymnothorax polyuranodon is one species that is known to be present in freshwater in the Indo-Pacific, but its life history is unknown. One way to evaluate what types of habitats are used by fishes is to determine the ratio of strontium (Sr) to calcium (Ca) in their otoliths, because this can show if they have used freshwater or saltwater environments. To evaluate the patterns of freshwater use by this unusual species of marine eel, the otolith Sr/Ca ratios of four G. polyuranodon (275–344 mm) caught in a freshwater stream of Fiji were analyzed. The consistently low Sr/Ca values (0–4) indicated upstream movement after settlement and freshwater or estuarine residence of all four individuals. These eels did not appear to have entered freshwater just for a short time period, which is consistent with other reports that this species is present in estuarine and freshwater habitats. This suggests that G. polyuranodon may be a catadromous species of marine eel. The similarities and differences between the life histories of anguillid eels and the few marine eels that have evolved the ability to invade freshwater habitats is discussed in relation to the evolutionary origin of diadromy in anguilliform fishes that originated in the marine environment.     

Reconstructing larval growth and habitat use in an amphidromous goby using otolith increments and microchemistry

High‐resolution analysis of growth increments, trace element chemistry and oxygen isotope ratios (δ¹⁸O) in otoliths were combined to assess larval and post‐larval habitat use and growth of Awaous stamineus, an amphidromous goby native to Hawai‘i. Otolith increment widths indicate that all individuals experience a brief period of rapid growth during early life as larvae and that the duration of this growth anomaly is negatively correlated with larval duration. A protracted high‐growth period early in larval life is associated with a lower ratio of Sr:Ca, which may reflect low salinity conditions in nearshore habitats. A distinct shift in δ¹⁸O (range: 4–5‰) is closely associated with the metamorphic mark in otoliths, indicating that larval metamorphosis occurs promptly upon return to fresh water. Strontium and other trace elements are not as tightly coupled to the metamorphosis mark, but confirm the marine‐to‐freshwater transition. Integration of microstructural and microchemical approaches reveals that larvae vary substantially in growth rate, possibly in association with habitat differences. Although time and financial costs make it difficult to achieve large sample sizes, present results show that examining even a small number of individuals can lead to novel inferences about early life history in diadromous fishes and illustrates the value of integrating analyses.     

Regional variation in otolith Sr:Ca ratios of African longfinned eel Anguilla mossambica and mottled eel Anguilla marmorata: a challenge to the classic tool for reconstructing migratory histories of fishes

Otolith Sr:Ca ratios of the African longfinned eel Anguilla mossambica and giant mottled eel Anguilla marmorata from nine freshwater sites in four rivers of South Africa were analysed to reconstruct their migratory life histories between freshwater and saltwater habitats. For A. mossambica, the Sr:Ca ratios in the otolith edge differed significantly among rivers and had large effect sizes, but did not differ among sites within a river. Otolith Sr:Ca ratios did not differ among rivers for A. marmorata. When rivers were pooled, the edge Sr:Ca ratios of A. mossambica were not significantly different from those of A. marmorata. According to the river-specific critical Sr:Ca ratio distinguishing freshwater from saltwater residence, most A. mossambica and A. marmorata had saltwater habitat experience after settlement in fresh water. This was primarily during their elver stage or early in the yellow eel stage. During the middle and late yellow eel stage, freshwater residency was preferred and only sporadic visits were made to saltwater habitats. The data also suggest that regional variations in otolith Sr:Ca ratios affect the critical Sr:Ca value and are a challenge for the reconstruction of migratory life histories that should be explicitly considered to avoid bias and uncertainty.     

Habitat residency and movement patterns of Centropomus parallelus juveniles in a subtropical estuarine complex

Sixty Centropomus parallelus juveniles were collected in March 2013 in two locations (Tromomó and Guaraguaçu) inside the Paranaguá estuarine complex, southern Brazil. The habitat residency and movement patterns of the individuals were inferred from Sr:Ca ratios and age recorded in the otoliths. Data suggest that the species spawns preferentially in brackish areas mainly from October to January, and that growth rate during the early juvenile stage could be influenced by environmental salinity. Furthermore, the data also show that C. parallelus can occupy diverse salinity habitats and migrate among marine, brackish and freshwater areas within the Paranaguá estuarine complex, showing a high environmental plasticity and adaptation.     

洞庭湖中是否存在溯河洄游型刀鲚

利用电子探针微区分析(EPMA)技术, 分析了2018年8月采自湖南省岳阳市洞庭湖畔水产市场2尾刀鲚(Coilia nasus)干制个体的矢耳石中锶和钙微化学特征。定量线分析结果显示, 2尾样本的耳石锶钙比值(Sr/Ca×1000)波动显著, 从耳石核心到边缘可分别出现对应淡水生境的锶钙比值<3的低值区, 对应于半咸水生境的3—7高值区和再次为对应于淡水生境<3的低值区。面分布分析结果更直观显示两尾刀鲚个体的耳石从核心到边缘对应于不同盐度生境的锶含量变化图谱, 即从耳石核心到边缘可分别出现对应于淡水生境的蓝色区域, 对应长江口外半咸水生境的黄绿色区域和再次对应于淡水生境的蓝色区域。这种淡水-半咸水-淡水的生境转换显示出了上述个体的典型溯河洄游生活史“履历”。研究显示目前距长江河口约1400 km的洞庭湖中仍可能存在溯河洄游型刀鲚个体。     

Using otolith chemical and structural analysis to investigate reservoir habitat use by juvenile Chinook salmon Oncorhynchus tshawytscha

Isotopic composition of ⁸⁷Sr:⁸⁶Sr and natural elemental tracers (Sr, Ba, Mg, Mn and Ca) were quantified from otoliths in juvenile and adult Chinook salmon Oncorhynchus tshawytscha to assess the ability of otolith microchemistry and microstructure to reconstruct juvenile O. tshawytscha rearing habitat and growth. Daily increments were measured to assess relative growth between natal rearing habitats. Otolith microchemistry was able to resolve juvenile habitat use between reservoir and natal tributary rearing habitats (within headwater basins), but not among catchments. Results suggest that 90% (n = 18) of sampled non‐hatchery adults returning to the Middle Fork Willamette River were reared in a reservoir and 10% (n = 2) in natal tributary habitat upstream from the reservoir. Juveniles collected in reservoirs had higher growth rates than juveniles reared in natal streams. The results demonstrate the utility of otolith microchemistry and microstructure to distinguish among rearing habitats, including habitats in highly altered systems.     

Extrinsic factors affecting otolith chemistry – implications for interpreting migration patterns in a diadromous fish

Much has been revealed about fish migration, including diadromous behaviour, through the use of otolith chemistry. Manipulative experiments assist with unravelling information on otolith chemical composition and incorporation thereby answering specific questions on diadromous movements. In this study, a laboratory-based experiment was used to determine the relative and interactive effects of salinity and water temperature on the composition of three key elements (Sr, Ba and Mg) within the otolith of a catadromous fish, Percalates novemaculeata, endemic to south-eastern Australia. Otolith incorporation of Sr and Ba was positively related to ambient water concentration, whereas Mg incorporation was not. Sr and Ba increased and decreased significantly across salinity gradients, respectively, with minor positive effects of temperature also being detected. Salinity and temperature interacted to significantly affect the elemental concentration ratios for Ba: Ca in otoliths. Discrimination between fresh water and marine environments shows promise for interpreting P. novemaculeata residency based on these elements alone. However, deciphering finer scale movements within estuarine environments may be difficult. Our data highlights the importance of multifactorial validation experiments and suggests complementary use of multiple approaches for unravelling species-specific patterns of fish movement and habitat use.     

Microchemical provenancing of prey remains in cormorant pellets reveals the use of diverse foraging grounds

Piscivorous birds in aquatic ecosystems exert predation pressure on fish populations. But the site-specific impact on fish populations, including stocked and commercially used fish species, remains disputed. One of the key questions for the management of piscivorous birds and fish is determining the origin of prey and thus which fish populations are targeted by the birds. We addressed this question by provenancing otoliths (earstones) of fish obtained from regurgitated pellets of piscivorous birds by otolith microchemistry analysis. We retrieved otoliths from regurgitated pellets of great cormorants (Phalacrocorax carbo sinensis) collected every 2 weeks for 2 years from breeding and roosting colonies at Chiemsee in Bavaria, Germany, and classified them according to family or species. We collected water samples from Chiemsee and potential surrounding foraging grounds. We measured the strontium (Sr) ⁸⁷Sr/⁸⁶Sr isotope ratio and Sr mass fraction of water and otoliths using (laser ablation) inductively coupled plasma-mass spectrometry. We assigned otoliths from regurgitated pellets to habitat clusters of origin by comparing the Sr isotopic and elemental composition of otoliths and waterbodies. In 36% of cormorant pellets collected at Chiemsee, prey was assigned to waterbodies distinct from Chiemsee. Furthermore, cormorants used different foraging sites during 1 day. Microchemical provenancing of prey remains can contribute to identifying foraging sites of piscivorous birds and to what extend the birds switched among foraging sites.     

Flexible Life History Strategies of Ninespine Sticklebacks,Genus <Emphasis Type="BoldItalic">Pungitius</Emphasis>

Synopsis We studied the life histories of the ninespine sticklebacks, Pungitius pungitius and Pungitius tymensis, collected from Japanese freshwater and brackish (sea) water habitats by examining the strontium (Sr) and calcium (Ca) concentrations in their otoliths. The Sr:Ca ratios in the otoliths changed with the salinity of the habitat regardless of identification as freshwater or brackish water type based on morphological characteristics. The ninespine sticklebacks living in a freshwater environment showed consistently low Sr:Ca ratios throughout the otolith. These samples were identified as a standard freshwater type. In contrast, all freshwater-type fishes collected from the intertidal zone showed higher otolith Sr:Ca ratios than those in the standard freshwater type, and the ratios fluctuated with the growth phase. All brackish water-type fishes collected in the intertidal zone showed the highest otolith Sr:Ca ratio throughout the otolith. In the present study, besides the two representative life history types of P. pungitius, i.e., freshwater and brackish water life history types, other sticklebacks had an anadromous life history type. These findings clearly indicate that the ninespine stickleback has a flexible migration strategy with a high degree of behavioral plasticity and an ability to utilize the full range of salinity in its life history.     

Reconstructing habitat use of Coilia mystus and Coilia ectenes of the Yangtze River estuary, and of Coilia ectenes of Taihu Lake, based on otolith strontium and calcium

The habitat use and migratory patterns of Osbeck's grenadier anchovy Coilia mystus in the Yangtze estuary and the estuarine tapertail anchovy Coilia ectenes from the Yangtze estuary and Taihu Lake, China, were studied by examining the environmental signatures of strontium and calcium in their otoliths using electron probe microanalysis. The results indicated that Taihu C. ectenes utilizes only freshwater habitats, whereas the habitat use patterns of Yangtze C. ectenes and C. mystus were much more flexible, apparently varying among fresh, brackish and marine areas. The present study suggests that the spawning populations of Yangtze C. ectenes and C. mystus in the Yangtze estuary consist of individuals with different migration histories, and individuals of these two Yangtze Coilia species seem to use a variety of different habitats during the non-spawning seasons.     

Facultative amphidromy involving estuaries in an annual amphidromous fish from a subtropical marginal range

Variation in the life cycle of diadromous fishes can be explained by differential food availability between marine and freshwater habitats, since migration is often interpreted as a mechanism for exploiting food resources. Theoretically, a migration pattern of mainly remaining in freshwater occurs in tropical and subtropical habitats where fluvial productivity possibly exceeds marine productivity. However, in Yakugachi River, Amami-Oshima Island, southern Japan, low nutrient concentrations in the river suggest that food availability is limited for the subtropical Ryukyu-ayu Plecoglossus altivelis ryukyuensis. Since Ryukyu-ayu is an amphidromous fish that mainly grows in rivers after spending 2 months in the sea, limited food availability in rivers would force this species to migrate to other habitats with better food availability. Otolith increment and Sr:Ca analyses of 48 adult Ryukyu-ayu collected from the Yakugachi River revealed that all individuals visited estuaries more than three times after moving upstream. Although the specific growth rates of this species in the river had no correlation with the salinity profile in the fluvial period, this movement may be an adaptive choice because the salinity profile significantly affected the body size at maturity. Our results highlighted individual-based variations in amphidromous migration for utilizing estuaries, which could be explained by relatively higher productivity in estuarine than in freshwater and marine habitats.     

Trace element and strontium isotopic analysis of Gulf Sturgeon fin rays to assess habitat use

Trace element and ⁸⁷Sr/⁸⁶Sr isotope analyses of fish pectoral fin rays offer non-destructive methods for determining habitat use. In this study, water and fin ray samples were analyzed for Gulf Sturgeon Acipenser oxyrinchus desotoi from the Choctawhatchee River Basin (FL and AL, USA) and compared with reference samples from Atlantic Sturgeon A. o. oxyrinchus held at controlled salinities (0, 10, 33 ppt). Samples were analyzed using inductively coupled plasma mass spectrometry, with a multi-collector for ⁸⁷Sr/⁸⁶Sr. In water, Sr, Ba, Mn and Zn differed between freshwater and saline habitats, with increases in Sr and decreases in Ba, Mn and Zn. ⁸⁷Sr/⁸⁶Sr decreased upstream to downstream with lowest values in saline habitats. In the reference study, water trace element concentrations and ⁸⁷Sr/⁸⁶Sr corresponded to those in pectoral fin rays. ⁸⁷Sr/⁸⁶Sr was higher in pectoral fin ray than water, due to influence of diet, which differed with salinity. In wild fish, trace elements in pectoral fin rays indicated freshwater emigration to saline habitats primarily occurred in the second to third growth zone with some heterogeneity in the population (4% <0.3 years, 39% 0.5–1.3 years, 39% 1.5–2.3 years, 17% 2.5–3.3 years). Analyses of ⁸⁷Sr/⁸⁶Sr indicated initial locations of Gulf Sturgeon were in the middle river, with few fish in the upper or lower river. Most (74%) juvenile Gulf Sturgeon utilized more than one river region prior to freshwater emigration and 48% moved upstream temporarily based on increased ⁸⁷Sr/⁸⁶Sr. After initial freshwater emigration, fish utilized lower-river to saline habitats. Collectively, these studies demonstrate the usefulness of trace element and ⁸⁷Sr/⁸⁶Sr analyses in sturgeon pectoral fin rays.     

Environmental signature in the otolith elemental fingerprint of the tapertail anchovy, Coilia mystus, from the Changjiang estuary, China

The ontogenetic patterns of habitat use and the migratory history of the tapertail anchovy, Coilia mystus, collected in the Changjiang estuary around Chongming Island, China, were studied by examining the environmental signature in the otolith strontium (Sr) and calcium (Ca) fingerprints using electron probe microanalysis (EPMA). Our results suggest that the migration strategy of C. mystus is much more flexible than supposed in the literature to date. The spring spawning population of C. mystus from the studied area was found to consist of individuals with different migration histories. Although the tapertail anchovy seems to be an anadromous fish that spawns and hatches in a freshwater habitat, it can also use a freshwater environment in non‐spawning seasons. The otolith EPMA of the elemental fingerprint (Sr x‐ray maps and Sr : Ca ratios) is an environmental indicator that can be applied to the migratory ecology of other important diadromous species in China.     

Unravelling the life-history patterns and habitat preferences of the Japanese eel (Anguilla japonica) in the Pearl River,China

Eels have fascinated biologists for centuries due to their amazing long-distance migrations between freshwater habitats and very distant ocean spawning areas. The migratory life histories of the Japanese eel, Anguilla japonica, in the waters of south China are not very clear despite its ecological importance, and the need for fishery regulation and management. In this study, strontium (Sr) and calcium (Ca) microchemical profiles of the otoliths of silver eels were measured by X-ray electron probe microanalysis based on data collected from different habitats (including freshwater and brackish habitats), in the large subtropical Pearl River. The corresponding habitat preference characteristics were further analysed using redundancy analysis (RDA). A total of 195 Japanese eels were collected over 6 years. The collected individuals ranged from 180 to 771 mm in total length and from 8 to 612 g in body weight. Two-dimensional pictures of the Sr:Ca concentrations in otoliths revealed that the A. japonica in the Pearl River are almost entirely river eels, spending the majority of their lives in fresh water without exposure to salt water, while the catadromous migration time has delayed about 1 month in the Pearl River estuary in the past 20 years. RDA analysis further indicated that juveniles and adults preferred water with high salinity and high tide levels. Youth preferred habitats with high river fractals. Our findings contribute to a growing body of evidence showing that the eels are extremely scarce currently and conservation measures against them are imminent, including the protection of brackish and freshwater areas where they live in south China.