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Tenualosa ilisha was found recently in the Perak River in western Peninsular Malaysia. Molecular phylogenetic and haplotype network analyses suggest that T. ilisha has two genetically distinct populations/groups: (i) Peninsular Malaysia (Malaysia population), and (ii) Peninsular Malaysia, Thailand, India and Bangladesh (Indian Ocean population). The results also suggest that the T ilisha population in Peninsular Malaysia is genetically heterogeneous with a typical anadromous migration pattern.

Rationale Otolith strontium:calcium (Sr:Ca) ratios have been extensively used to study fish migration across a river, estuary and ocean at each life stage. However, otolith Sr:Ca ratios only explain partial variations in salinity and quantitative reconstruction of salinity history of fishes remains a challenge. Using a euryhaline fish Lateolabrax japonicus as an example, this study demonstrated an isotopic method of quantitatively reconstructing the salinity histories and habitat uses of the fish. Methods Otolith oxygen stable isotopic ratios (δ18 Ooto values) were measured using a mass spectrometer for subsamples sequentially milled from the otolith core to the edge, and otolith Sr:Ca ratios were measured by an electron probe microanalyzer for the comparison. The mean water temperature within the studied area in the time period of each milled subsample was estimated from the Copernicus database. Based on an isotopic fractionation equation, each δ18 Ooto value and water temperature pair determined the water δ18 O value, which was then converted into salinity using published linear equations or an equation that was newly generated for this study. Results Individual fish clearly revealed different preferences in habitat use. The retrospectively reconstructed salinity history indicated that most L. japonicus inhabit and can spawn in seawater as well as in brackish estuaries. Few fish lived in freshwater during the young stage based on the analysis of δ18 Ooto profiles; however, otolith Sr:Ca ratios can only reveal freshwater residence and cannot distinguish residence in brackish water or seawater. Conclusions This study demonstrated that otolith temporal microstructure and δ18 Ooto values are well-suited approaches for quantitative reconstruction of salinity histories of the fish. This method can improve the understanding of the habitat uses of other fishes inhabiting diverse habitats among the rivers, estuaries and oceans.

The feeding behavior of the giant ambush-predator “Bobbit worm” (Eunice aphroditois) is spectacular. They hide in their burrows until they explode upwards grabbing unsuspecting prey with a snap of their powerful jaws. The still living prey are then pulled into the sediment for consumption. Although predatory polychaetes have existed since the early Paleozoic, their bodies comprise mainly soft tissue, resulting in a very incomplete fossil record, and virtually nothing is known about their burrows and behavior beneath the seafloor. Here we use morphological, sedimentological, and geochemical data from Miocene strata in northeast Taiwan to erect a new ichnogenus, Pennichnus. This trace fossil consists of an up to 2 m long, 2–3 cm in diameter, L-shaped burrow with distinct feather-like structures around the upper shaft. A comparison of Pennichnus to biological analogs strongly suggests that this new ichnogenus is associated with ambush-predatory worms that lived about 20 million years ago.

Anguillid eels of the genus Anguilla, which have a unique catadromous life history, are widely distributed across many parts of the world. However, little research has been conducted on the behavioural mechanisms of habitat segregation between sympatric species in tropical anguillid eels. To understand the ecological and behavioural mechanisms involved in the life history and migration of tropical anguillid eels, strontium (Sr):calcium (Ca) ratios were examined in otoliths of A. bengalensis bengalensis (41 specimens) and A. bicolor bicolor (130 specimens) collected from ten rivers in northwestern Peninsular Malaysia. The otolith Sr:Ca ratios revealed different habitat use between the two species. The broad range of otolith Sr:Ca ratios and habitat shift found in A. bicolor bicolor suggested that its habitat utilization was opportunistic in environments of varying salinity. A. bicolor bicolor prefers to live in the midstream to downstream areas with tidal influences. A. bengalensis bengalensis, however, was found to only reside in freshwater environments throughout their continental growth. A. bengalensis bengalensis tends to live in upstream area with no tidal influence. Their habitat use, migratory history, and habitat distribution indicate that habitat segregation occurs between the two species, leading to the different habitat preferences in tropical river systems.

Determination of the geographic boundaries that define a stock is a key challenge in fisheries management. Acquiring an understanding of spatial distribution and migration patterns of pikeperch Sander lucioperca populations in the south-eastern Baltic Sea region would help to delineate the stocks and contribute towards sustainable exploitation of the population. In this study, habitat use and migratory behaviour of S. lucioperca were inferred from otolith Sr:Ca ratios obtained via an Electron Probe Micro-Analyzer. S. lucioperca collected from the inland Kaunas City water reservoir, where downstream migration is precluded by a hydro-power plant, consistently showed a freshwater signature in their otoliths throughout their life history. In contrast, those collected from the Baltic Sea and Curonian Lagoon in Lithuania, and the lower reaches of the Daugava River in Latvia had ratios indicative of movement between fresh and brackish water habitats. Although S. lucioperca can inhabit brackish water bodies, they predominantly reside and reproduce in freshwater systems. It is thought that periodic post-spawning migration to the sea during spring is related to better foraging conditions and perhaps as a strategy for shedding ectoparasites. Currently, catch quotas apply only to those S. lucioperca taken from the Curonian Lagoon and not the Baltic Sea. Frequent migration between freshwater habitats and the Baltic Sea suggest that S. lucioperca stocks from a given river/estuary system should be considered as a single management unit despite individual differences in migratory behaviour. The extent to which migration into the Baltic Sea could potentially compromise the effectiveness of catch quotas was the main issue which initiated this study.

The relationships between pelagic larval duration (PLD) and geographic distribution patterns or population genetic structures of fishes remain obscure and highly variable among species. To further understand the early life history of the tidepool snake morayUropterygius micropterusand the potential relationship between PLD and population genetic structure of this species, otolith microstructure and population genetics based on concatenated mtDNA sequence (cytochromeband cytochrome oxidase subunit I, 1,336 bp) were analyzed for 195 specimens collected from eight locations around the southern Ryukyu Islands, Taiwan, and the central Philippines. Eels with longer PLD and lower otolith growth rates were observed at relatively higher latitudes with lower water temperatures (54.6 ± 7.7 days and 1.28 ± 0.16 µm day−1on Ishigaki Island, Japan, vs. 43.9 ± 4.9 days and 1.60 ± 0.19 µm day−1on Badian, the Philippines), suggesting that leptocephali grew faster and had shortened pelagic periods in warmer waters. Meanwhile, the eels along the southwest coast of Taiwan had relatively longer PLD (57.9 ± 10.5 days), which might be associated with the more complex ocean current systems compared to their counterparts collected along the east coast of Taiwan (52.6 ± 8.0 days). However, the southwestern and eastern Taiwan groups had similar otolith growth rates (1.33 ± 0.19 µm day−1vs. 1.36 ± 0.16 µm day−1). Despite the intergroup variation in PLD, genetic analysis revealed fluent gene flow among the tidepool snake morays in the study regions, implying that intraspecies PLD variation had a weak effect on genetic structure. The leptocephalus stage might have ensured the widespread gene flow among the study areas and leptocephalus growth was likely influenced by regional water temperature.

We investigated the clinical effectiveness of subcutaneous (SC) administration of recombinant feline interferon-omega (rFeIFN-ω) at a dose of 1 M unit (MU)/kg body weight (bw) for the treatment of feline chronic gingivitis-stomatitis (FCGS) in cats infected with feline calicivirus (FCV). Among the 17 cats used in this study, there were 13 FCV-positive cats (FCVI group), which were subcutaneously injected with rFeIFN-ω. The remaining four FCV-positive cats (FCVC group) were treated with SC corticosteroid. SC injection of rFeIFN-ω was given once daily on days 1, 2, 3, 7, 8, 9, 14, and 21. Corticosteroid was subcutaneously injected at a dose of 1.0 mg/kg bw, at the same intervals as rFeIFN-ω. Clinical symptoms (salivation, pain at opening the mouth, halitosis, mandibular lymphadenopathy, and all four symptoms combined [defined as “total clinical symptoms”]) and stomatitis (the degree and extent of inflammation, bleeding from the lesion, and all three items combined [defined as “total stomatitis”]) were scored on days 0, 7, 14, 21, and 28. FCV RNAs was quantified by real-time polymerase chain reaction and the percent increase in viral copy numbers was calculated using the values on days 0 and 28. In the FCVI group, significant differences were observed in the score for clinical symptom (salivation) score and in the total clinical symptom score within the group (P = 0.018 and 0.008, respectively). Significant differences within the group were also observed in the scores for the degree and extent of inflammation in stomatitis and in the total stomatitis score (P = 0.003, 0.007, and 0.003, respectively). The total score, defined as the clinical score plus the stomatitis score, was on days 7, 14, 21 and 28 than on day 0 (p = 0.006, .0003, 0.002 and 0.002, respectively). In the FCVI group, significant difference was observed between on days 0 and on 21 (p = 0.023). The percentage change in the number of polymerase chain reaction cycles required to amplify the viral RNA was positive (indicating viral reduction) in the FCVI group, but was negative in the FCVC group. These results demonstrate that SC administration of rFeIFN-ω under the current protocol improves stomatitis by inhibiting FCV proliferation in FCV-positive cats with FCGS.

The migratory life histories were investigated for 30 African longfinned eels (Anguilla mossambica) of the yellow eel stage from the Maningory River, East Madagascar using otolith Sr:Ca ratios and parasite composition. The Sr:Ca ratio patterns of nine eels resembled those of A. mossambica in South Africa and of other anguillids and were classified as “typical” Sr:Ca ratio pattern eels. Typical pattern eels were further classified into freshwater residents (five eels) and habitat shifters (four eels) depending on whether they spent their first several years in saltwater. An unique Sr:Ca ratio pattern was found in 21 eels whereby the Sr:Ca ratios in the yellow eel stage increased drastically to extremely high values of 10 to 30 × 10−3 which have not been observed for any Elopomorpha species. These individuals were categorized as “elevated” Sr:Ca ratio pattern eels. The elevated pattern eels were characterized by significantly older ages, lower somatic growth rates, a much higher intensity and abundance of the gastrointestinal parasite Heliconema africanum and a lower abundance of the swim bladder parasite Anguillicoloides papernai. The extremely elevated Sr:Ca ratios of the yellow eel stage result most likely from inhabiting freshwater areas characterized by a high strontium concentration because the ratios far exceeded the known values for Elopomorpha species. Cyclical lows in the otolith Sr:Ca ratios of the elevated type eels corresponded to annuli, revealing either an overwintering movement pattern between habitats with different Sr:Ca ratios, or a seasonal change in the ambient environment.

This study using tuna otoliths as working standards established a high lateral resolution and precision analysis to measure δ 18 O 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 δ 18 O otolith values as a proxy of water temperature. Dramatic increases up to 5–6‰ in δ 18 O 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. δ 18 O 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 δ 18 O otolith profiles as the result of Cs 2+ 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 δ 18 O 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.

The Japanese eel Anguilla japonica is an important aquaculture fish species in the East Asian countries of Japan, China, Korea, and Taiwan. All glass eel fry are captured from the wild and understanding the recruitment patterns of the glass eel is important. The larvae of A. japonica are passively transported to the East Asian Continental Shelf by the North Equatorial Current, the Kuroshio, the Kuroshio intrusion currents, and coastal currents. In each location, recruitment time is diverse: It is November in Taiwan and April in the Yalu River. How the glass eels reach recruitment areas remains poorly understood. Here, we combine information from larval ages based on otolith increments, simulated drifting paths on the East Asian Continental Shelf, and main fishing seasons in each location of East Asia. We identify five main recruitment blocks: (1) The main Kuroshio, (2) The Taiwan Strait Warm Current, (3) The Taiwan Warm Current, (4) The Yellow Sea Warm Current and (5) The branch of Yellow Sea Warm Current. The counted age of the glass eels is significantly underestimated for the later recruits, possibly due to the cessation of the otolith edge growth under low water temperatures. This study clarifies the eel&rsquo, s larval characteristics and transport mechanisms in the East Asia Continental Shelf, providing important information for its recruitment dynamics in the marine stage.

Linear discriminant analysis (LDA) and habitat discrimination critical value (HDCV) methods are alternatives for evaluating the migratory history, such as the proportion of residence in freshwater (%fwr) and estuarine/marine habitats, of individual American eels (Anguilla rostrata) via the analysis of otolith Sr:Ca data. The LDA process requires Sr:Ca data from both freshwater and estuarine/marine groups while the mean + cSD method requires only a freshwater standard. For the two group (freshwater, estuarine/marine), single predictor variable (Sr:Ca) case, the LDA process defaults to Fisher's linear discriminant where the HDCV equals the average of the group mean Sr:Ca values. The difference between freshwater and estuarine resident eel mean otolith Sr:Ca values, based on 13 published studies and the current study (n = 14), decreased with increasing freshwater group otolith mean (r = 0.80, p

Lagoons and estuaries are transitional waters (TW), saline in character but substantially influenced by freshwater (FW) flows and the most productive habitat after upwelling areas. The European eel, Anguilla anguilla, is a typical inhabitant of these habitats and a target of important lagoon fisheries since ancient times. Notwithstanding this, in the Mediterranean region, where the numerous coastal lagoons are the most eligible habitat for this species, eel habitat use and growth, which display a high inter-individual variability, are poorly studied. To gain knowledge about the migratory behaviour and the relative growth history of this species in TW in the Mediterranean area, the otolith Sr/Ca ratios of 56 individuals were analysed. The study sampling sites were two typical coastal lagoon environments, Caprolace lagoon and Lesina lagoon, located, respectively on the Tyrrhenian and the Adriatic coast of Italy, and the Tiber River (TR) estuary, in Central Italy. Otolith Sr/Ca profiles revealed that in all the sites, the resident contingent is substantial, while the proportion of nomad eels, or habitat inter-shifter, was different among sites. Mean annual grow rate of resident eels is higher in productive environments (TR and lagoon of Lesina), while in Caprolace lagoon, an oligotrophic lagoon, resident eels grow slower. The observed patterns of habitat use and growth seem linked to local ecological conditions: facultative movements of eels with a nomadic behaviour seem affected by food availability rather than by the salinity gradient. This consideration supports the hypothesis that the facultative catadromous migration of European eel in Mediterranean TW, and the trophic shifts that this species exhibits, depend primarily on the productivity of the environment rather than on its salinity gradient.

By combining the spatio-temporal distribution of fish abundance and their size structure, and a detailed lifetime Sr/Ca ratio analysis in their otoliths, this study delineates the estuarine use and the movement patterns of seven sympatrical occurring adult mullets in the Tatu Creek estuary, central western Taiwan. In the estuary Mugil cephalus are the most dominant species, whereas Liza subviridis, Liza macrolepis and Liza haematocheilus are common, and Liza affinis, Liza dussumieri and Valamugil seheli are rare. They have adapted a size-related salinity preference. A mean Sr/Ca ratio of (7.5–10.2) × 10−3 in the otolith cores demonstrated that all seven mullet species spawned in the sea. After recruiting to the estuary (mean ratios of (3.6–6.4) × 10−3 at the estuarine check in the otoliths), the ratios fluctuate between (0.1–3.5) × 10−3 and (9.5–19.5) × 10−3 indicating that the mullet shared a common movement between marine and brackish waters and probably even freshwater habitats. However, the profiles fluctuated substantially among individuals. There was high intra-specific variation among M. cephalus and L. subviridis, intermediate intra-specific variation among L. macrolepis and L. affinis, and relatively little among L. haematocheilus, L. dussumieri and V. seheli. Persistent residency in high or low saline environments was found to vary among species, and the extent of their catadromy is discussed.

European eel Anguilla anguilla migratory behaviour during continental life is still unclear due to the multiplicity of aquatic environments colonised by the species. In the Camargue area (NW Mediterranean), eel colonisation of the Fumemorte canal, a freshwater habitat that com- municates only with a vast brackish ecosystem (the Vaccares lagoon), offers a rare opportunity to test for freshwater habitat residence during continental life. To this end, both laser ablation inductively coupled plasma mass spectrometry and electron probe micro-analysis were used to measure chronological variations of strontium concentrations (Sr:Ca) in the otoliths of 58 silver eels captured in the canal between 1997 and 2007. Comparing mean Sr:Ca ratios measured on oto- lith edges with the 2 analytical methods indicated that they provide comparable measurement accuracies. Linear discriminant analysis (LDA), based on the otolith Sr:Ca values corresponding to the initial entrance of the fish into the brackish ecosystem and their final capture in the freshwater canal, allowed successful discrimination of the 2 habitats and reconstruction of migratory history for all individuals. Six different migratory behaviours were identified. Eels that entered the fresh- water canal did so either directly (67%) or after 1 to 2 yr spent in the lagoon (33%), with a subse- quent majority of freshwater residents (55%) until their silvering. These results indicate the value of LDA for reconstructing habitat use during continental life using Sr:Ca ratios. They confirm the occurrence of freshwater residence during continental life in European eels, even in Mediter- ranean continental areas where brackish environments are predominant. This observed seden- tary behaviour has implications for eel population management and conservation.

Otolith δ 18 O and δ 13 C of six species grenadiers were analyzed to reconstruct the historical residing depths and metabolic activity. During the larval to juvenile stage, Spicomacrurus kuronumai , Hymenocephalus lethonemus , and Bathygadus nipponicus gradually migrated from the mixed layer to the thermocline downward over a vertical distance >140 m after which they moved within a narrow vertical range for the remaining life. The downward migration distance was less than 65 m during the larval stage of Hymenocephalus sp. and Coryphaenoides acrolepis , which showed a second descent period from the thermocline to deeper water as juveniles. Coryphaenoides marginatus stayed at the lower thermocline during the larval stage and the juveniles migrated downward in a relative short distance around 100 m and settled in deeper water (>600 m), followed by irregular movements over a vertical range of about 200 m during juvenile and adult stages. The otolith δ 13 C profile suggested that fishes ( S. kuronumai , H. lethonemus , and B. nipponicus ) with a longer migration distance had a higher metabolic rate in their early life-history stages than in the later stages. However, the metabolic rate did not vary for those fishes ( H. sp., C. acrolepis and C. marginatus ) living within a narrow vertical range during their larval to adult stages. The otolith microchemistry suggested that ontogenetic downward migration was an important strategy for grenadiers linking the life stages between pelagic larvae and benthic settlement. Furthermore, the migration timing and distance for the pelagic larvae varied between species and habitats. S. kuronumai , H. lethonemus , H. sp. and C. acrolepis might metamorphose and settle at the same time, B. nipponicus metamorphosed during migration and C. marginatus migrated as juveniles.

To understand their migratory environmental history and habitat use in Mexican waters, 31 striped mullet Mugil cephalus and 40 white mullet M. curema (family Mugilidae) were collected from three habitats (Tamiahua Lagoon, and Cazones and Tecolutla estuaries, Veracruz) in the western Gulf of Mexico. Sr:Ca ratios in otoliths of mullets were measured from the primordium to the otolith edge with an electron probe microanalyzer. Mullets were aged by counting the otolith annuli. The patterns in mullet otolith Sr:Ca ratio differed among habitats and between species, indicating that habitat use and migratory behavior differed between species. Variability in otolith Sr:Ca ratios was consistent with that of the environment inhabited by the mullets. They showed a more diverse euryhaline preference than the simple catadromous behavior expected. The Sr:Ca ratios in the core region of the otolith indicated that mullets can spawn in both offshore waters and nearshore brackish waters, or that spawning may occur in offshore waters and that onshore currents/winds carry the fertilized eggs into onshore waters where salinities are lower. Peak Sr:Ca ratios corresponded to otolith annulus deposition, which validated the difference in timing of seaward spawning migration between striped mullet (winter) and white mullet (spring).

To examine the relationship between freshwater entry and otolith annular structures, a total of 113 naturally recruited European eels Anguilla anguilla from Lithuania and Latvia that entered fresh water at least once were collected. In some individuals (8.3-11.3%), the first freshwater entry coincided with a dark check that was distinctly different from neighbouring annuli. In most individuals (81.7-84.9%), the first freshwater entry occurred on rings and increments indistinguishable from other annuli. For the remaining individuals (3.8-10%), the first freshwater entry did not correspond to any otolith ring, band or annulus. According to recent evidence, the observed high correspondence between the first freshwater entry and otolith annuli was more likely due to the movement into fresh water during winter when the annulus was deposited, rather than stress resulting from habitat change. Consequently, the age estimation based on otoliths might be less influenced by this habitat change during the yellow eel stage.

Growth of American eels, Anguilla rostrata (Lesueur), sampled in saltwater bays and adjacent freshwater ponds in the southern Gulf of St. Lawrence, Canada, was estimated by otolith annulus back-calculation, and related to salinity of growth habitat as determined using otolith strontium:calcium ratios. Eels resident in salt water grew on average in length 2.2 times faster than freshwater residents and increased in weight 5.3 times faster than freshwater residents. Previous findings of superior American eel growth rate in brackish water are thus extended to full-strength salt water. Freshwater residents took an estimated 2.4 times longer than saltwater residents to reach the silver eel stage. The extended time to maturation of eels reared in fresh water increases their vulnerability to fisheries and to bio-accumulation of contaminants. Freshwater eel populations should be managed more conservatively than those in saline waters. The increase in eel growth with salinity suggests that rearing in brackish or salt water could improve growth rates in eel aquaculture.

The Pacific tarpon Megalops cyprinoides is a euryhaline fish that is widely distributed in offshore waters of the tropical and subtropical Indo-Pacific Ocean. It spawns in offshore waters and the larvae drift with tidal currents before recruiting to estuarine nursery areas at ~25 to 30 d post- hatching. Young Pacific tarpon are found in coastal waters, rivers, estuaries, inner bays, and man- grove areas. However, their age-specific migratory behavior and habitat selection are still unclear. The strontium (Sr) concentration in seawater is ~100× higher than in fresh water, and the Sr:Ca ratios in fish otoliths are positively correlated with ambient salinity and are thus widely used in studying fish migration between seawater and freshwater environments. To understand the migratory envi- ronmental history of the Pacific tarpon, otolith Sr:Ca ratios of leptocephali collected from the estuary and sub-adults collected from both Tadu Creek and offshore waters on the west coast of Taiwan were examined using an electron probe microanalyzer (EPMA). Fish were aged by counting otolith annuli. The Sr:Ca ratios in leptocephalus otoliths were high (7.4 ± 0.03 × 10 -3 ). However, the patterns of oto- lith Sr:Ca ratios in 1 to 3 yr old sub-adults were diverse and can be divided into 3 types: largely brack- ishwater residents, largely freshwater residents, and vagrants between brackish and fresh waters. Fish older than 4 to 5 yr all return to the marine environment. This study demonstrates that habitat selection in the Pacific tarpon after the leptocephalus stage is facultative and that their migratory environmental history can be reconstructed from otolith Sr:Ca ratios.

We examined the effects of water temperature, salinity and feeding regime on lepto- cephalus metamorphosis, and on daily growth increment deposition and strontium:calcium (Sr:Ca) ratios of otoliths in the Pacific tarpon Megalops cyprinoides. The tarpons at the pre-metamorphic lep- tocephalus stage (SI) were collected in the estuary, marked with tetracycline and then reared for 18 and 30 d in 2 independent experiments with different temperatures (20, 25 and 30°C), salinities (0, 10 and 35 psu) and feeding regimes (fed and starved)—environmental conditions that the fish may experience in the wild. Temporal changes in Sr:Ca ratios from the primordium to the otolith edge of the reared tarpon were examined with an electron probe microanalyzer. At the optimal temperature (25°C) in Expt I, the leptocephalus completed metamorphosis (SII and SIII) after ca. 2 wk irrespective of being fed or starved and reared in low (10 psu) or high (35 psu) salinity, although both somatic and otolith growth rates were lower in starved than in fed groups. However, in Expt II, metamorphosis was delayed at SII when the larvae were reared close to winter water temperature (20°C), and the duration of metamorphosis was reduced to less than 12 d when the leptocephalus was reared at 30°C. This indicated that the rate of metamorphosis was significantly influenced by water temperature. Meanwhile, the drastic change in Sr:Ca ratios and increment width in the otolith of the leptocephalus during metamorphosis was also significantly different among different temperatures, salinities and feeding regimes in different degrees. The present study indicated that these factors significantly influenced the metamorphosis rate of the leptocephalus and subsequently their daily growth increment deposition and Sr:Ca ratios in otolith.

The presence of vaterite mosaic in the aragonite otoliths of anguillid eel otoliths is read- ily determined by reflected light microscopy after etching with EDTA. The prevalence and intensity of vaterite inclusions in sagittal otoliths of 165 American eels Anguilla rostrata were examined in relation to latitude and migratory history. Prevalence varied significantly (range: 21.3 to 58.8%) among 3 rivers in Atlantic Canada and increased non-linearly with latitude. Habitat differences among sites may confound the relation because most (48%) vateritic otoliths came from eels with a history of estuarine residence and inter-habitat migration. The limited number of sites, restricted geo- graphic range and modest otolith sample sizes makes premature any conclusion that the prevalence of vateritic otoliths in American eels increases with latitude. The frequency of vaterite inclusions in otoliths (n = 63) was low (1 to 6) and did not vary among sites. Vaterite intensity was low (

Most fish otoliths are composed of aragonite, but occasionally they form a mosaic with its polymorph, vaterite. We found an unusual mosaic structure of extremely low strontium to calcium (Sr/Ca) ratio in the aragonite otoliths of European eels Anguilla anguilla. The mosaic crystal structure in the eel otolith appeared opaque under reflected light after EDTA etching and was confirmed to be a vaterite crystal by Raman spectroscopy. Analysis by laser ablation inductively coupled plasma mass spectrometry indicated that the elements sodium (Na), strontium (Sr) and barium (Ba) were lower in abundance and magnesium (Mg) and manganese (Mn) were higher in the vaterite than in the aragonite. Otoliths with vaterite inclusions were found in 48.1% of the 108 eels examined. If the mosaic vaterite in otoliths is not identified and avoided, the migratory environmental history of fish could be misidentified when otolith elemental signatures are used as biological tracers.

Analyses of otolith strontium:calcium (Sr:Ca) ratios in 162 American eels Anguilla ros- trata of the St. Jean River watershed in eastern Canada demonstrated the co-existence of 6 migratory patterns, including freshwater and brackish water residence, and the predominance of an amphidro- mous migratory behavior. We tested the hypothesis that the choice of a particular life-history tactic may be controlled by a conditional strategy with status-dependent selection. This prediction was not supported because migratory patterns did not vary as a function of individual size, age and/or sex of eels prior to migration. However, we demonstrated that the utilisation of the estuarine brackish envi- ronment, more productive than the freshwater river and lake, resulted in a higher growth rate. Fresh- water yellow eels, the typical catadromous tactic, were rare and experienced lower growth rates. Moreover, we suggest that freshwater overwintering periods are not recorded in otoliths owing to the cold winter climate that occurs at northern latitudes. Thus, Sr:Ca ratio patterns that show brackish residence may in some cases hide a seasonal migration between fresh and brackish habitats.

The use of Sr:Ca ratios in fish otoliths to reconstruct historical patterns of fish movement between aquatic habitats of different salinity ranges (fresh, estuarine, marine) can be extended to evaluate the frequency and duration of inter-habitat movements. However, the proportion of otolith growth in a habitat does not necessarily equal the proportion of time spent in that habitat and depends on the difference between otolith growth rates within each habitat. For inter-habitat migrant yellow American eels Anguilla rostrata from the East River, Nova Scotia, the mean proportion of res- idence time in freshwater slightly (2.9%), but significantly, exceeded the proportion of otolith growth in freshwater, but the magnitude of the effect was small and perhaps of little practical consequence. Although the observed effect magnitude was small, where large differences in otolith growth rates occur among habitats or with ontogeny, habitat-specific growth rates should be considered for detailed examinations of inter-habitat migration and residency. A simple habitat residence model provides results consistent with the observed data. For inter-habitat migrants, the mean difference between 2 possible methods of estimating the proportion of fish growth in freshwater from micro- probe otolith Sr:Ca ratio transects was small (2.2%). Singleton outliers of eel otolith Sr:Ca ratios may create difficulties in evaluating the frequency and duration of inter-habitat movements that remain to be resolved.

Although otolith Strontium (Sr)/calcium (Ca) ratios have been widely used to reconstruct the past salinity environmental history of anguillid eels, factors affecting the Sr/Ca ratios in otoliths are incompletely understood. Japanese Eel (Anguilla japonica) elvers (mean length 54.7±2.1 mm) were collected in the estuary during their upstream migration and reared at 5 different salinities (0, 5, 15, 25, and 35 psu) and 3 types of feeding conditions (formulated feed, tubifex, and starvation) for 30 days to evaluate the effects of salinity and diets on otolith Sr/Ca ratios. Ca and Sr concentrations in the ambient water significantly increased with salinity (SAL) as [Ca] water=15.50SAL�5.56, and [Sr] water=0.21SAL+0.03, respectively. Sr/Ca ratios in otoliths increased with salinity (SAL) of the rearing water as [(Sr/Ca)×1000] otolith=0.091SAL+3.790. In diets, Sr/Ca ratios were 4 times higher in tubifex than in formulated feed. However, in otoliths, ANOVA indicated that Sr/Ca ratios did not differ significantly between groups fed on tubifex or formulated feed (p=0.118). Otolith Sr/Ca ratios were negatively correlated with fish growth rates while the growth rates differed significantly among rearing conditions with different salinities and diets. Partition coefficients of the Sr/Ca ratios from ambient water to fish tissues and otoliths significantly increased with salinity. The Sr/Ca ratios of Japanese Eel otoliths thus were positively correlated with the ambient salinity and decreased with increasing fish growth rate, but was not affected by fish diet.

Otolith Sr:Ca ratios were examined to evaluate the contribution of the stocked eel Anguilla anguilla elvers, which have been stocked in Lithuanian waters and mixed with naturally recruited eels for several decades, to the native eel population. Stocked eels were identified by the freshwater signature (Sr:Ca ratios 3·23 x 10 -3 ) after the glass eel stage. Of 108 eels analysed, 21 eels had otolith Sr:Ca ratio profiles consistent with stocking while 87 showed patterns of natural recruitment. The ages of naturally recruited eels arriving in Lithuanian fresh waters varied from 1 to 10 years, with a mean ± S.D. age of 5.2 ± 2·1 years. Eels from the inland Lake Baluosai were all freshwater residents of stocked origin. Stocked eels, however, accounted for only 20% of the eels from the Curonian Lagoon and 2% of eels sampled in Baltic coastal waters. This finding does not support the hypothesis that the eel fishery in the Curonian Lagoon depends mostly on stocking.

The temporal patterns in the otolith Sr:Ca ratios of yellow-phase American eels Anguilla rostrata from the East River, Chester, on the Atlantic coast of Nova Scotia, Canada, indicated variable patterns of migration between river and estuarine/marine waters. About 29% of yellow eels (n = 107) had a history of migration between river and estuary other than their initial entry. Male and sexually undifferentiated eels, potentially mostly male, comprised 96% of the sample and females comprised 4%, but the proportions of migratory and non-migratory eels did not vary by sex or development stage. Most migrants (81%) made only 1 round trip, while 19% made 2 round trips. Yellow eels with a migratory history were patchily distributed, with most (84%) concentrated 9.4 km upriver. About 36% of yellow eels showed a check (false annulus) corresponding with a habitat transition from estuary to river, while for 16% of yellow eels the check occurred during the initial freshwater entry as an elver. Otolith checks not associated with a habitat transition occurred for 22% of yellow eels, primarily at intermediate ages. Mean lengths-at-age were higher for eels with a history of primarily estuarine growth than for eels of primarily freshwater growth, as were annual growth rates (26.6 ver- sus 21.7 mm yr -1 ).

The blind cavefish, Milyeringa veritas, inhabits an anchialine system, effectively a groundwater estuary, in which salinity varies between fresh and seawater at different locations and depths. Owing to the inaccessible habitat and the threatened status of the cavefish it is hard to obtain the biological information needed for their management. This paper explores the utility of otolith Sr:Ca ratio in elucidating cavefish biology. The mean Sr:Ca ratio of the water inhabited by the cavefish is correlated with both the TDS (total dissolved solids) of the habitat and with the Sr:Ca ratio of the sagittal otolith of the cavefish inhabiting that site. Mean values of Sr:Ca in otoliths suggest some cavefish inhabit sea or brackish waters while others remain in freshwater. Some individuals appear to move between waters of very different TDS at various stages but there is no consistency in the direction or apparent TDS range of the water bodies inhabited which indicates that the cavefish utilise the different water bodies opportunistically. Residual analyses indicate clear and routine changes in the TDS of the water occupied at various phases of growth, irrespective of the TDS at which the cavefish were sampled. Annular markings are present in some otoliths but they cannot be related to likely periodicities in the subterranean environment.

Sr/Ca ratios in the otoliths of 2 marine eels, the moray eel Gymnothorax reticularis and the pike eel Muraenesox cinereus, decreased greatly when the crystal structure of otolith daily growth increments changed from concentric to radiate form. This decrease is proposed to correspond to the timing of metamorphosis from the leptocephalus to glass eel stage, similar to freshwater eels (Anguilla spp.). In freshwater eels, ontogenetic and habitat shifts might influence the decrease. How- ever, marine eels do not migrate into freshwater after completing metamorphosis, so the decrease in these species must be the result of metamorphosis-related physiological, rather than environmental, effects. The mean age at metamorphosis of the moray eels was significantly lower than that of pike eels, and both were lower than that of freshwater eels (p < 0.05). Consequently, the spawning ground of marine eels is presumed to be closer than that of freshwater eels to the continental shelf.

Otolith Sr:Ca ratios were used to determine migrational histories of American eels Anguilla rostrata in an eastern Canadian coastal stream that is impounded at the head of tide. American eels sampled from the estuary were smaller (41.0 ± 9.3 cm (mean ± SD), N = 324) and younger (ages 1–4) than those in a freshwater pond (65.0 ± 7.5 cm, N = 265; ages 11–26) 2.5 km upstream of the head of tide. The Sr:Ca ratios permitted identification of three migrational contingents. In 7 of 13 (54%) American eels taken in the estuary, Sr:Ca ratios declined to 4.76 ± 1.22 × 10−3, indicating direct migration from the sea to estuarine settlement sites. In the 6 other American eels (46%), Sr:Ca ratios indicated an approach to freshwater soon after arrival in continental waters, followed by settlement in the estuary where ratios stabilized at 4.50 ± 1.23 × 10−3. In all 15 American eels sampled from the freshwater pond, Sr:Ca ratios indicated settlement in freshwater in the elver year and no subsequent change of habit...

We used an electron probe microanalyzer (EPMA) to determine the migratory environ- mental history of the catadromous grey mullet Mugil cephalus from the Sr:Ca ratios in otoliths of 10 newly recruited juveniles collected from estuaries and 30 adults collected from estuaries, nearshore (coastal waters and bay) and offshore, in the adjacent waters off Taiwan. Mean (±SD) Sr:Ca ratios at the edges of adult otoliths increased significantly from 6.5 ± 0.9 × 10 -3 in estuaries and nearshore waters to 8.9 ± 1.4 × 10 -3 in offshore waters (p < 0.01), corresponding to increasing ambi- ent salinity from estuaries and nearshore to offshore waters. The mean Sr:Ca ratios decreased sig- nificantly from the core (11.2 ± 1.2 × 10 -3 ) to the otolith edge (6.2 ± 1.4 × 10 -3 ) in juvenile otoliths (p < 0.001). The mullet generally spawned offshore and recruited to the estuary at the juvenile stage; therefore, these data support the use of Sr:Ca ratios in otoliths to reconstruct the past salinity history of the mullet. A life-history scan of the otolith Sr:Ca ratios indicated that the migratory environmen- tal history of the mullet beyond the juvenile stage consists of 2 types. In Type 1 mullet, Sr:Ca ratios range between 4.0 × 10 -3 and 13.9 × 10 -3 , indicating that they migrated between estuary and offshore waters but rarely entered the freshwater habitat. In Type 2 mullet, the Sr:Ca ratios decreased to a minimum value of 0.4 × 10 -3 , indicating that the mullet migrated to a freshwater habitat. Most mullet beyond the juvenile stage migrated from estuary to offshore waters, but a few mullet less than 2 yr old may have migrated into a freshwater habitat. Most mullet collected nearshore and offshore were of Type 1, while those collected from the estuaries were a mixture of Types 1 and 2. The mullet spawning stock consisted mainly of Type 1 fish. The growth rates of the mullet were similar for Types 1 and 2. The migratory patterns of the mullet were more divergent than indicated by previous reports of their catadromous behavior.

The strontium (Sr) and calcium (Ca) concentrations in the otoliths of the Japanese eels Anguilla japonica collected from China, Japan and Taiwan were measured by electron probe micro-analyzer. The Sr/Ca ratios indicated that the eels beyond elver stage can be classified into three types of migratory contingents. Type 1 (seawater), the Sr/Ca ratios from approximately 150 Am from primordium to edge of the otolith maintained at the level of approximately 4–10x, indicating that the eel after elver stage stayed in sea water until the silver eel stage. Type 2 (freshwater), the ratios were lower than 4x, indicating that the eel stayed in freshwater from elver stage to the silver eel stags. Type 3 (estuarine), the ratios fluctuated between those of Types 1 and 2, indicating that eel migrated between freshwater and sea water before the silver stage. The estuarine contingents constituted the majority of the eel population and grew faster than the freshwater contingents. D 2003 Elsevier Science B.V. All rights reserved.

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.

Ozdilek, Sukran Yalcin/0000-0001-8264-7606; WOS: 000287920800013 PubMed: 21366578 Otolith Sr:Ca ratios from 32 of 34 European eel Anguilla anguilla collected from three freshwater sites in the River Asi, southern Turkey, indicated that they were resident in fresh water without apparent exposure to salt water since the elver stage. The Sr:Ca ratio criterion indicative of residence in fresh water was more than twice that of values from other European countries. Otolith Sr:Ca ratios of A. anguilla from fresh waters can vary among regions, possibly reflecting regional-specific water chemistry. Hence, the use of Sr:Ca ratios determined in one region to interpret results from a different region might lead to misclassification of migratory life-history types.

Otolith strontium:calcium ratios were used to trace lifetime movements of American eels (Anguilla rostrata) captured in salt-water bays and adjoining freshwater ponds in Prince Edward Island, Canada. Eels were classified into migratory contingents based on their movement patterns. A pond with a pool-and-weir salmonid fishway and a pond drained by a low-gradient channel contained eels that had entered freshwater at all ages, but a pond with a 2.2 m vertical spillway contained only eels that had entered freshwater in the elver year. Salt-water residents were the dominant migratory contingent in salt-water bays (85% of 39), which overturns the paradigm of obligate catadromy for this species. Freshwater residency was the sole pattern found in the pond with the vertical spillway (100% of 12) and the majority contingent in the pond with the low-gradient channel (54% of 24). Inter-habitat shifting was the dominant migratory contingent in eels sampled from the pond with the pool-and-weir fishway (85% of 20). Resident eels were established in salt- and freshwater habitats by the year after their arrival in continental waters. Eels that shifted between habitats increased their rate of inter-habitat shifting with age. The high degree of plasticity in habitat use found in this study is consistent with worldwide Anguillid patterns as revealed by Sr:Ca.

The differences in geographical distribution between Anguilla reinhardtii and A. australis on the eastern coast of Australia can be understood by comparing otolith growth increments and microchemistry, the ages between species of the eels at metamorphosis from leptocephalus to glass eels and the ages of glass eels at estuarine arrival. The ages at metamorphosis were determined from where the increment width dramatically increased and the Sr/Ca ratio dropped. The mean age (± s.d.) of A. reinhardtii (n = 176) at metamorphosis was 144.5 ± 12.2 days and at estuarine arrival was182.7 ± 16.3 days. For A. australis (n = 150) it was 173.7 ± 20.5 days and 229.2 ± 29.4 days, respectively. The differences in age between species were significantly larger than the annual and seasonal variations within species. Australian eels are believed to spawn in the tropical oceans and larval eels drift in the South Equatorial Current to eastern Australia. The younger ages at estuarine arrival of A. reinhardtii suggest that the spawning grounds of this species lie closer to Australia than those of A. australis. In addition, the mean total length at recruitment of A. reinhardtii (49.9 ± 2.0 mm) was significantly smaller than for A. australis (54.6 ± 5.4 mm) (t = 3.8, P < 0.01). However, the growth rates of A. reinhardtii (0.25 ± 0.02 mm/d) were significantly faster than for A. australis (0.23 ± 0.022 mm/d)(t = 7.6, P < 0.01). The smaller sizes of A. reinhardtii at recruitment were likely due to the shorter marine larval period and faster growth rate compared with A. australis. The duration of the marine larval period and growth rate may be the principal factors in determining the geographical distribution of both A. reinhardtii, which tend to occur in tropical-subtropical waters, and A. australis, which predominate in more temperate waters. Extra keywords: otolith, early life history. M la t ar nt Jao et W A nd Y.

The environmental history of American eels Anguilla rostrata from the East River, Nova Scotia, was investigated by electron microprobe analysis of the Sr:Ca ratio along transects of the eel otolith. The mean (±SD) Sr:Ca ratio in the otoliths of juvenile American eels was 5.42 × 10 -3 ± 1.22 × 10 -3 at the elver check and decreased to 2.38 × 10 -3 ± 0.99 × 10 -3 at the first annulus for eels that migrated directly into the river but increased to 7.28 × 10 -3 ± 1.09 × 10 -3 for eels that had remained in the estuary for 1 yr or more before entering the river. At the otolith edge, Sr:Ca ratios of 4.0 × 10 -3 or less indicated freshwater residence and ratios of 5.0 × 10 -3 or more indicated estuarine residence. Four distinct but interrelated behavioural groups were identified by the temporal changes in Sr:Ca ratios in their otoliths: (1) entrance into freshwater as an elver, (2) coastal or estuarine residence for 1y r or more before entering freshwater, and, after entering freshwater, (3) continuous freshwater res- idence until the silver eel stage and (4) freshwater residence for 1 yr or more before engaging in peri- odic, seasonal movements between estuary and freshwater until the silver eel stage. Small (< 70 mm total length), highly pigmented elvers that arrived early in the elver run were confirmed as slow growing age-1 juvenile eels. Juvenile eels that remained 1 yr or more in the estuary before entering the river contributed to the production of silver eels to a relatively greater extent than did elvers that entered the river during the year of continental arrival.

To understand the migratory behavior and habitat use of the Japanese eel Anguilla japonica in the Kaoping River, SW Taiwan, the temporal changes of strontium (Sr) and calcium (Ca) contents in otoliths of the eels in combination with age data were examined by wavelength dispersive X-ray spectrometry with an electron probe microanalyzer. Ages of the eel were determined by the annulus mark in their otolith. The pattern of the Sr:Ca ratios in the otoliths, before the elver stage, was similar among all specimens. Post-elver stage Sr:Ca ratios indicated that the eels experienced different salinity histories in their growth phase yellow stage. The mean (±SD) Sr:Ca ratios in otoliths beyond elver check of the 6 yellow eels from the freshwater middle reach were 1.8 ± 0.2 x 10 -3 with a maximum value of 3.73 x 10 -3 . Sr:Ca ratios of less than 4 x 10-3 were used to discriminate the freshwater from seawater resident eels. Eels from the lower reach of the river were classified into 3 types: (1) freshwater contingents, Sr:Ca ratio