Frequency of vateritic otoliths and potential consequences for marine survival in hatchery-reared Atlantic salmon

Otoliths are inner-ear structures of all teleost fish with functional importance for hearing and balance. The otoliths usually consist of aragonite, a polymorph of calcium carbonate, but may also take the form partly or entirely of vaterite, a different polymorph of calcium carbonate. Vateritic otoliths occur sporadically in wild fish, but with a higher frequency in hatchery-reared fish. Abnormal otoliths have direct consequences for the inner-ear functions of fish and may be a symptom of environmental stress. In this study, the authors assess the differences in the frequency of abnormal otoliths and degree of abnormality (% vaterite) for different groups of hatchery-reared Atlantic salmon (Salmo salar) smolt and adults. The groups differed in parental broodstock origin (number of generations in hatchery) and treatment temperature. Smolt from the same groups were also released to complete their ocean migration. The otoliths of the returning and recaptured adults were subsequently extracted to assess the difference in frequency and degree of abnormality between the adults and the smolt from corresponding groups. Return rate varied among groups (0.2%–2.6%). The frequency of vateritic otoliths was high (11.4%–64.4%) and differed among smolt groups. The lowest return rates corresponded with the highest frequency of abnormal otoliths for the groups, suggesting that abnormal otoliths may have negative consequences for marine survival. Furthermore, indications of an effect of fast growth on the formation of abnormal otoliths were found for only one of the experimental groups, and for none of the groups after correcting for Type 1 error. This contradicts previous reports, suggesting rapid growth as the main cause of abnormal otoliths. Adult return rates were generally low, but abnormal otoliths were common, with high coverage (% vaterite).     

Larval history of glass eels, Anguilla anguilla (Linné, 1758) in migration in coastal and estuary areas (Adour, Gulf of Gascogne) as revealed by otolith test]

The embryonic past of glass eels was studied from the interpretation of microstructures registered on otoliths. The aim of this work is to put in evidence possible seasonal modifications of the growth of otoliths so that differences between otoliths of glass eels caught off marine and estuarine environment. So during the season 1999-2000, from November till March, otolith sampling was realised in the southwestern part of France, in an estuarine and coastal zone. We observed a spatial and temporal evolution of proportions of the three various types of otoliths taken into account. Glass eels sampled at sea sometimes have a mark on their otoliths indicating the transition in the estuary, especially at the end of the fishing season. Measures of growth marks of otoliths showed that there were no seasonal differences during phases of the transoceanic migration and the crossing of the continental shelf. The radius of otoliths of glass eels sampled at sea was significantly smaller than those sampled in estuary. These results translated homogeneous environmental modifications met by the various larvae groups during the oceanic crossing and during the principal migration season as well as a turn over of these groups during the transition between marine and continental environment.     

An assessment of otoliths,dorsal spines and scales to age the long‐finned gurnard,Lepidotrigla argus,Ogilby, 1910 (Family: Triglidae)

Sagittal otoliths, dorsal spines and scales were critically assessed as structures to potentially determine the age of the long‐finned gurnard, Lepidotrigla argus. Counts were made of opaque growth increments and a readability score was assigned to each structure. Comparisons of growth increment counts were made between structures and between readings. All three structures showed some degree of readability and quantifiable growth increments, but this varied within fishes and between structures. Initial results showed that whole otoliths were more suitable to determine age estimates than dorsal spines and scales. Scales were considered unsuitable due to between reading ageing bias, variation in age estimates between structures, low precision and poor readability for this species. Dorsal spines showed evidence of loss of growth increments due to hollowing of the vascular core, which resulted in underestimation of older individuals in comparison to whole otoliths. Further analysis showed that growth increment counts from whole otoliths were lower for older individuals in comparison to sectioned otoliths. It is suggested that this is because of decreased clarity of growth increments towards the outer margin of whole otoliths in older individuals; this problem was not present with sectioned otoliths. It was concluded that sectioned otoliths were a more suitable structure from which to estimate age of L. argus than were whole otoliths, dorsal spines and/or scales.     

Effect of DNA extraction on ageing success in coral trout (Plectropomus leopardus) otoliths

To maximize the information commonly collected from otoliths, the effect of DNA extraction on the estimation of age with otoliths was evaluated by comparing sagittal otolith samples from common coral trout ( Plectropomus leopardus ) for clarity and ageing discrepancies in DNA-extracted and untreated control otoliths. The DNA extraction process had no significant effect, indicating that archived otoliths can be used as a source of DNA while retaining their utility for age estimation.     

Otolith microstructure pattern in Oreochromis niloticus (L.)

The microstructure of otoliths from young and old Oreochromis niloticus (L.) were studied. Otoliths were prepared histologically except for those from newly hatched fish. Hatching results in the formation of a check in the otoliths, which appeared 1 day later. Other checks are rare in juvenile otoliths but common in adult otoliths. Faint and non-daily increments were observed within the hatching check. After hatching, increments were deposited daily. Sub-daily increments were faint and narrow, they were present in the area along the dorso-ventral axis of the otolith but did not continue into the lateral region. Discontinuous zones in the medial area appeared different from those in the lateral area. New growth centres were not only found in the juvenile fish otoliths, but also in adult fish otoliths.     

Fish otoliths: do sizes correlate with taxonomic group,habitat and/or luminescence?

Otoliths are dense structures in the ears of fishes that function in hearing and gravity perception. Otolith (sagitta) diameters, as percentages of standard length (% SL), are calculated for 247 marine fish species in 147 families and compared by taxonomic group (usually order), habitat and presence or absence of luminescence. Otolith sizes range from 0.4-31.4 mm and 0.08-11.2% SL. The eel and spiny eel orders Anguilliformes and Notacanthiformes have small to very small otoliths, as do the triggerfish order Tetraodontiformes, pipefish order Gasterosteiformes, billfish suborder Scombroidei and many of the dragonfish order Stomiiformes. The soldierfish order Beryciformes has moderate to very large otoliths. The perch order Perciformes has a wide range of otolith sizes but most have small to moderate otoliths 2-5% SL. Only 16 out of the 247 species have the relatively largest otoliths, over 7% SL. Seven out of these 16 species are also luminous from a variety of habitats. Luminous species have slightly to much larger otoliths than non-luminous species in the same family Both beryciforms and luminous fishes live in low-light environments, where acute colour vision is probably impossible. Most fishes of the epipelagic surface waters have very small otoliths, perhaps due to background noise and/or excessive movement of heavy otoliths in rough seas. Bathypelagic species usually have small otoliths and regressed or absent swimbladders. Other habitats have species with a range of otolith sizes. While the relationship between hearing ability and otolith length is unknown, at least some groups with modified swim-bladders have larger otoliths, which may be associated with more acute hearing.     

A comparison of different structures and methods for estimating age of northern-form Dolly Varden <Emphasis Type="Italic">Salvelinus malma malma</Emphasis> from the Canadian Arctic

Assessment of anadromous northern-form Dolly Varden Salvelinus malma malma in the Western Canadian Arctic requires reliable methods for estimating ages. Additionally, conservation efforts warrant determining whether fin rays provide a non-lethal alternative to otoliths. Precision and bias of whole and sectioned otoliths, and sectioned pectoral and pelvic fin rays were examined. Two age readers with different levels of experience ageing this species read each structure three times. Coefficient of variation (CV) was calculated to measure precision, and age bias plots were created for each method of preparation/structure within and between readers. The experienced reader demonstrated the highest precision with sectioned otoliths (CV = 1.6 %) followed by whole otoliths (CV = 4.2 %) while pectoral and pelvic fins were the lowest, CV = 7.7 % and 7.5 %, respectively. The age bias plot showed little difference between whole and sectioned otoliths, although greater imprecision/bias was evident for whole otoliths at age ≥9. Compared to otoliths, fin rays produced younger age estimates starting at 5 years; however, pelvic fins were more biased towards younger estimates than pectoral fins. The less experienced reader had greater inconsistencies, tending to overage younger and underage older samples for all methods compared to the more experienced reader, underscoring the importance of experience when estimating age for this species. We conclude that both types of fin rays are a poor non-lethal alternative to otoliths for fish ≥5 years and recommend an experienced ager could use whole otoliths up to age 8 and sectioned otoliths for fish ≥9 years (>500 mm fork length).     

Validation of age readings from otoliths of juvenile roundnose grenadier, Coryphaenoides rupestris, a deep-water macrourid fish

Validation of the ageing of deep-water fish is difficult and there are only a few instances where the rings on the otoliths have been shown to be laid down annually. Roundnose grenadier have been fished commercially in the North Atlantic since the 1960s and the adult fish have frequently been aged by counting the rings in otoliths or scales. All the ageing was done on the assumption that the rings in the otoliths or scales were annual. Between 1975 and 1992, the Scottish Association for Marine Science carried out seasonal trawling surveys in the Rockall Trough using a fine-meshed trawl, and collected otoliths from a wide size range of roundnose grenadier. An examination of the growing edge of otoliths from juvenile fish from these collections suggests that the rings in the otoliths are laid down annually. The broader, opaque zones which represent the growth phase were dominant between September and March. The thinner, hyaline zones were dominant between April and July. The apparent delay in the growth phase compared with most shallow-water species is discussed in relation to the availability of mesopelagic prey.     

Selenium detected in fish otoliths: a novel tracer for a polluted lake?

To test if otoliths can be used to track fish migration in polluted areas, fish sampled from Onondaga Lake, heavily polluted with mercury, were used in an assay to determine whether mercury was detectable in the fishes’ otoliths using synchrotron-based scanning X-ray fluorescence microscopy (SXFM). Mercury was undetectable, but selenium, rarely reported in otoliths and known as a physiological antagonist to mercury, was. Strontium was also present but appeared to be taken up independently of selenium, and thus these serve as independent biogeochemical markers. Both selenium and mercury were detected in fish tissues, but selenium was below levels considered toxic. Selenium was low in otoliths of fishes collected in nearby Oneida Lake. Synoptic surveys of water chemistry revealed that Se is regionally highest in Onondaga Lake and in particular its main inlet, Onondaga Creek. SXFM appears to be a sensitive method for detecting selenium in otoliths.     

Suitability of glycerin-preserved otoliths for age validation using bomb radiocarbon

The effect of glycerin storage on the bomb radiocarbon content of otoliths was determined experimentally. Storage in either pre- or post-bomb glycerin had no detectable effect on the bomb radiocarbon content of either pre- or post-bomb otoliths. Therefore bomb-dated age validation studies need not be restricted to freshly-collected samples or dry otoliths, implying that the large numbers of glycerin-archived otoliths around the world are suitable for age validation studies using bomb radiocarbon.     

Crystalline otoliths in teleosts: Comparisons between hatchery and wild coho salmon (<Emphasis Type="Italic">Oncorhynchus kisutch</Emphasis>) in the Strait of Georgia

Otoliths, or ‘ear stones’, are calcium carbonate structures found in all vertebrates. In teleosts, they have a number of sensory functions, including hearing. Daily growth increments of these structures have permitted advanced age and population studies of teleosts. Whereas ‘normal’ otoliths are composed of crystals imbedded within a protein matrix as aragonite, a ‘crystalline’ form of calcium carbonate termed vaterite is also found. A review of the otolith literature demonstrates a significant level of understanding of the structure and function of otoliths, but the cause for crystalline otolith structure remains speculative. Pairs of otoliths from hatchery and wild juvenile and adult coho salmon (Oncorhynchus kisutch) were examined visually for determination of otolith microstructure type. The vateritic or crystalline otoliths were found in much higher percentages in juvenile hatchery-reared coho salmon than in juvenile wild coho salmon, supporting previous studies. There did not seem to be any negative impact on size or survival. There was also no correlation between crystalline otoliths and premature maturation in coho males. A preliminary study of adult coho salmon returning to Big Qualicum and Chilliwack hatcheries showed even higher ratios of vateritic otoliths than observed in juveniles.     

Morphometry and composition of aragonite and vaterite otoliths of deformed laboratory reared juvenile herring from two populations

Vaterite otoliths were sampled from two reared populations (Celtic and Clyde Seas) of juvenile herring Clupea harengus. The crystallography, elemental composition and morphometry were analysed and compared with those of normal aragonite otoliths. The incidence of vaterite otoliths in the juveniles sampled (n = 601) ranged from 7·8% in the Clyde population to 13·9% in the Celtic Sea population, and was 5·5% in the small sample (n = 36) of wild adults examined. In all but one case fish had only one vaterite otolith; the corresponding otolith of the pair was completely aragonite. Although the majority of the juveniles sampled showed craniofacial deformities, there was no link between the skull or jaw malformation and the incidence of vaterite otoliths. All vaterite otoliths had an aragonite inner area, and vaterite deposition began sometime after the age of 90 days. The vaterite otoliths were larger and lighter than their corresponding aragonite partners, and were less dense as a consequence of the vaterite crystal structure. The vaterite areas of the otoliths were depleted in Sr, Na and K. Concentrations of Mn were higher in the vaterite areas. The transition between the aragonite inner areas and the vaterite areas was sharply delineated. Within a small spatial scale (20 μm³) in the vaterite areas, however, there was co‐precipitation of both vaterite and aragonite. The composition of the aragonite cores in the vaterite otoliths was the same as in the cores of the normal aragonite otoliths indicating that the composition of the aragonite cores did not seed the shift to vaterite. Vaterite is less dense than aragonite, yet the concentrations of Ca analysed with wavelength‐dispersive spectrometry (WDS) were the same between the two polymorphs, indicating that Ca concentrations measured with WDS are not a good indicator of hypermineralized zones with high mineral density. The asymmetry in density and size of the otoliths may cause disruptions of hearing and pressure sensitivity for individual fish with one vaterite otolith, however, the presence of vaterite otoliths did not seem to affect the growth of these laboratory reared juvenile herring.     

Morphology and microchemistry of abnormal otoliths in the ayu, <Emphasis Type="Italic">Plecoglossus altivelis</Emphasis>

The sagittal otolith morphology and microchemistry of reared juvenile ayu, Plecoglossus altivelis, were examined to describe the occurrence and microchemical characteristics of the abnormal otoliths in this species. Juvenile ayu (N = 31) were collected in June 2004 at three different locations, Wakayama, Kumamoto, and Biwa Lake in Japan, where they were being reared in freshwater aquaculture ponds after having been collected in the wild as larvae. Otolith abnormality was found in the sagittae of 26% (N = 8) of the individuals examined, of which five fish had abnormal otoliths only on one side, while the otolith on the other side was normal. Abnormal otoliths were more transparent and crystalline in appearance with irregular shapes compared to normal ones that were more opaque and less irregular. Abnormal otoliths were divided into two types, semi-abnormal (Type 1) with a normal part in the center, and fully-abnormal (Type 2) that were completely crystalline in appearance. The line transects and whole otolith concentration maps showed that the contents of Sr, Na and K were lower in the abnormal otolith regions compared to the normal ones, while those of Ca and S were almost constant in both. The appearance and microchemical properties of the abnormal ayu otoliths were similar to the abnormal otoliths in other species in which vaterite replaces the aragonite. Abnormal formation of otoliths occurred in ayu from Biwa Lake (30%) and Kumamoto (45%), while the Wakayama samples had no abnormality. The microchemical analyses of the normal and abnormal otoliths indicated that some abnormal otoliths had formed before the fish were captured and transferred to the hatchery, so the possible causes of otolith abnormality in ayu are discussed.     

Otoliths of sub-adult Lake Sturgeon Acipenser fulvescens contain aragonite and vaterite calcium carbonate polymorphs

In this study we quantified the percent CaCO₃ polymorph composition in otoliths of larval and juvenile Lake Sturgeon Acipenser fulvescens via X-ray microdiffraction. Sagittal otoliths of sub-adults were primarily composed of aragonite (> 90%) while the lapilli otoliths were 100% vaterite. This is the first time the presence of aragonite in otoliths has been reported in an acipenseriform and is surprising given that the ability to form aragonite otoliths was not thought to have evolved until the separation of teleost and holostean species from other Actinopterygian fishes (e.g., sturgeon, paddlefish, gar).     

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

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

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

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

Otolith resorption induced by anaerobic stress in the goldfish, Carassius auratus

To examine otolith resorption induced by anaerobic stress, ⁴⁵Ca-prelabelled goldfish, Carassius auratus , were kept in oxygen-deficient ambient water (O₂ < 0.5 .1 1⁻¹, 26° C) for 2 days and otoliths (asterisci) were analysed for ⁴⁵Ca retention. In a second experiment, fish were anaerobically stressed for 2 days, and then received a single intraperitoneal injection of ⁴⁵Ca. They were maintained under stress for one more day and killed to examine ⁴⁵Ca deposition in otoliths. Plasma was analysed for total and radioactive calcium. Otoliths (lapilli) were examined for stress-induced check formation by scanning electron microscopy (SEM).
Stress significantly reduced plasma calcium levels and the rate of calcium retention in otoliths, which was calculated from the ⁴⁵Ca specific activity of the plasma. On the other hand, the rate of calcium deposition in otoliths was the same in the stressed as in the control fish. SEM observation revealed that the applied stress resulted in a check formation in otoliths. These results indicate that a 48-h stress of oxygen deficiency induces calcium resorption in otoliths.     

Effect of somatic and otolith growth rate on stable isotopic composition of early juvenile cod (Gadus morhua L) otoliths

The relative amounts of the stable isotopes of carbon and oxygen in fish otoliths can be used to reveal the environmental history experienced by the fish. This requires that the relative amounts of the isotopes are deposited in equilibrium with the surrounding environment, or that the offset from this equilibrium is known and can be quantified. It is known that carbon isotopes in biogenic carbonates are a mixture of carbon from the seawater and metabolically derived carbon, but the effect of the somatic growth rate of the fish is still unclear. The possible effect of otolith growth rate and fractionation of both carbon and oxygen isotopes are also not established. We carried out a controlled laboratory experiment where we reared cod (Gadus morhua L.) larvae and early juveniles at two temperatures (6 and 10 °C) and generated different growth rates within each temperature by manipulation of prey levels. The otoliths of the resulting fish were analysed for carbon and oxygen isotopes. We found no effect of otolith precipitation rate on fractionation of either carbon or oxygen isotopes. However, there was a depletion of ¹³C in the otoliths of fish with elevated metabolism. The proportion of metabolically derived carbon in the otoliths was estimated to be 28-32%. Our results suggest that measurements of oxygen isotopes in otoliths can be a reliable tool to estimate ambient temperature since the oxygen isotopes seem to be deposited in the otoliths independently of kinetic and metabolic effects. Fractionation of carbon isotopes in otoliths on the other hand can give valuable insight into metabolism and feeding pattern of fish.     

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

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