Fish Inner Ear Research Articles

Diversity of fish otoliths from the Gulf of Mexico and the Caribbean Sea: Report on the first digital collection of fish otoliths (Actinopterygii) from the Atlantic region of Mexico

The Otolith Collection of Fishes from the Gulf of Mexico and Caribbean Sea was created with the objective to conserve and illustrate the diversity of these structures from species living in the Gulf of Mexico, the Caribbean Sea, and the freshwater and brackish systems of the Yucatan Peninsula, incorporating morphological descriptions and morphometric data. Otoliths, non-skeletal calcareous structures that develop in the inner ear of fish, are essential for balance and hearing. They have become pivotal tools for age and growth determination, population analysis, and ecological, trophic, and archaeological studies due to their resistance to degradation and the extensive information they provide about the environment and physiology of fish. The collection now includes otoliths from 214 species, representing 67 families, obtained through sampling campaigns and collaborations with local fishermen. The otoliths are extracted using techniques that ensure the integrity of the structures for later preservation. High-resolution images were obtained using optical and scanning electron microscopy, and these images are stored in an online database. This database facilitates research and teaching by providing public access to digital specimens and associated data. In addition to fostering academic development, this collection represents a significant step towards the creation of a national platform for otolith data analysis, aligned with international efforts to digitize biological collections. Despite the financial and logistical challenges involved in building and maintaining biological collections, this collection demonstrates its value as an essential resource for taxonomic, systematic, and ecological studies, as well as for biodiversity education and awareness. The online availability of the collection not only facilitates access to data but also promotes innovation and international collaboration in the study of fishes and their habitats.

Using teacher-student neural networks based on knowledge distillation to detect anomalous samples in the otolith images

Otoliths are small calcium carbonate structures found in the inner ear of fish and they, as one of important information carriers, are applied in diverse ecological fields. Otoliths are usually photographed and used to explore many unsolved biological and ecological questions. However, many anomalies may occur in the large volume of otolith image data due to natural or artificial consequences, which brings a huge bias to the aimed study and even misleading results. In this study, we first propose a specific definition of otolith anomalies and provide a dataset of otolith anomalies with Electrona carlsbergi, one of the most abundant species of lanternfishes, as the study subject. We modify a multiresolution knowledge distillation neural network model, the state-of-the-art anomaly detection model to a multiresolution knowledge distillation network model with asymmetric inputs, which uses grayscale maps to align the features of color maps in the feature space, to help improve otolith anomalies detection. Our fine-tuned anomaly detection network obtains a better anomaly identification performance with a Receiving Operating Characteristic Area Under the Curve value of 0.9843. Our result shown that multiresolution knowledge distillation networks can efficiently identify abnormal otolith image sample, which is of great importance for conducting otolith-based science.

Otolin-1, an otolith- and otoconia-related protein, controls calcium carbonate bioinspired mineralization

BackgroundOtoliths and otoconia are calcium carbonate biomineral structures that form in the inner ear of fish and humans, respectively. The formation of these structures is tightly linked to the formation of an organic matrix framework with otolin-1, a short collagen-like protein from the C1q family as one of its major constituents. MethodsIn this study, we examined the activity of recombinant otolin-1 originating from Danio rerio and Homo sapiens on calcium carbonate bioinspired mineralization with slow-diffusion method and performed crystals characterization with scanning electron microscopy, two-photon excited fluorescence microscopy, confocal laser scanning microscopy and micro-Raman spectroscopy. ResultsWe show that both proteins are embedded in the core of CaCO3 crystals that form through the slow-diffusion mineralization method. Both of them influence the morphology but do not change the polymorphic mineral phase. D.rerio otolin-1 also closely adheres to the crystal surface. General significanceThe results suggest, that otolin-1 is not a passive scaffold, but is directly involved in regulating the morphology of the resulting calcium carbonate biocrystals.

Morphologic, morphometric and contour shape variations of sagittal otoliths of Lepidorhombus spp. in the Aegean Sea.

Otoliths are calcareous anatomical structures in the inner ear of fishes, and they can be used in the discrimination of fish species and stocks due to their species-specific shape. Sagittal otoliths in the Lepidorhombus whiffiagonis and Lepidorhombus boscii, two flatfish species of the same genus distributed in the Aegean Sea, were compared using morphology, morphometry and contour shape analyses. Blind and eyed side otoliths of Lepidorhombus species were evaluated separately due to their morphological features and statistical differences in morphometric measurements (p < 0.05). Four analysis groups were formed: blind side otolith of L. whiffiagonis, eyed side otolith of L. whiffiagonis, blind side otolith of L. boscii and eyed side otolith of L. boscii. Morphometric differences in otolith shape were performed by canonical discriminant analysis, and the first discriminant axis explained 97.4% (Wilks λ=0.270) of the variance between the groups, and the second axis explained 2.3% (Wilks λ=0.620). Classification success between Lepidorhombus species based on CDA is 100%, and the overall CDA classification score between groups is 76.5%. The wavelet functions obtained in the contour analysis showed high variability in the anterior, posterior-dorsal, antero-dorsal and ventral regions of the otoliths among these four analysis groups. Consequently, it is thought that this study will contribute to the taxonomic classification of fish with the morphological and morphometric differences in the blind and eyed side otoliths and the analyses made with the high discrimination success obtained in Lepidorhombus species.

Does Vaterite Otolith Deformation Affect Post-Release Survival and Predation Susceptibility of Hatchery-Reared Juvenile Atlantic Salmon?

Sagittal otoliths are calcareous structures in the inner ear of fishes involved in hearing and balance. They are usually composed of aragonite; however, aragonite can be replaced by vaterite, a deformity which is more common in hatchery-reared than in wild fish. Vaterite growth may impair hearing and balance and affect important fitness-related behaviours such as predator avoidance. Captive rearing techniques that prevent hearing loss may have the potential to improve fish welfare and the success of restocking programmes. The aim of this study was to test the effect of structural tank enrichment on vaterite development in the otoliths of hatchery-reared juvenile Atlantic salmon Salmo salar, and to assess the effects of vaterite on immediate predation mortality and long-term survival after release into the wild. Fry were reared in a structurally enriched or in a conventional rearing environment and given otolith marks using alizarin during the egg stage to distinguish between the treatment groups. Otoliths were scrutinised for the presence and coverage of vaterite at 6, 13, and 16 weeks after start feeding, and the growth traits were measured for enriched and control fry when housed in tanks. In a subsequent field experiment, juveniles were released in the Rasdalen river (western Norway), and otoliths of enriched reared and control reared fry were scrutinised from samples collected immediately prior to release, from predator (trout Salmo trutta) stomachs 48 h after release and from recaptures from the river 2–3 months after release. Vaterite otoliths occurred as early as 6 weeks after start feeding in hatchery-reared S. salar. Vaterite occurrence and coverage increased with fish length. Enriched rearing had no direct effect on vaterite formation, but enriched reared fry grew slower than control fry. After release into the wild, fewer salmon fry with vaterite otoliths had been eaten by predators, and a higher proportion of fry with vaterite otoliths than those lacking vaterite were recaptured in the river 2–3 months after release. Contrary to expectations, this suggests that vaterite does not increase predation mortality nor reduce survival rates in the wild during the early life stages.

The inner ear proteome of fish.

The mechanisms that underpin the formation, growth and composition of otoliths, the biomineralized stones in the inner ear of fish, are largely unknown, as only a few fish inner ear proteins have been reported. Using a partial transcriptome for the inner ear of black bream (Acanthopagrusbutcheri), in conjunction with proteomic data, we discovered hundreds of previously unknown proteins in the otolith. This allowed us to develop hypotheses to explain the mechanisms of inorganic material supply and daily formation of growth bands. We further identified a likely protein mediator of crystal nucleation and an explanation for the apparent metabolic inertness of the otolith. Due to the formation of both daily and annual increments, otoliths are routinely employed as natural chronometers, being used for age and growth estimation, fisheries stock assessments, and the reconstruction of habitat use, movement, diet and the impacts of climate change. Our findings provide an unprecedented view of otolith molecular machinery, aiding in the interpretation of these essential archived data.

Analysis of cod otolith microchemistry by continuous line transects using LA-ICP-MS

Fish otoliths, also called ear stones or statoliths, are calcified structures functioning as movement and equilibrium indicators in the inner ear of fish (Fig. 1). From hatching to death these structures grow incrementally, with new material accreted daily (Pannella 1971) in successive layers of protein (1–8%, Degens et al. 1969) and calcium carbonate. The accretion rate of otoliths varies with fish growth, and in temperate species it is usually lowest during the winter season (Hüssy et al. 2010). This results in concentric growth resembling the ringed structure in trees (Fig. 1D), enabling the use of dendrochronological techniques to approximate the age and growth history of fish. During growth, certain elements are incorporated into the otolith structure, some associated with proteins and some with the calcium carbonate component (Thomas et al. 2017), supplying a valuable record of different aspects in fish life history and serving as a potential environmental record. Previous studies show that trace element and isotopic compositions of otoliths can be used as a proxy for reconstructing water chemistry, temperature and salinity (Patterson et al. 1993; Thorrold &amp; Shuttleworth 2000). Other studies demonstrate that elemental histories can be used to investigate fish spawning and migration patterns (e.g. Sturrock et al. 2012), and more recent studies use elements such as Zn, Cu and Mg as indicators of seasonality (Hüssy et al. 2016; Limburg et al. 2018). Combining this knowledge of elemental variation with the micro-beam capabilities of laser ablation inductively coupled plasma mass spectrometry (LAICPMS) turns otolith microchemistry into a powerful tool for studying important parameters fundamental for establishing modern, sustainable fisheries management policies (e.g. stock identification, migration, pollution indicators, spawning habitats, duration of larval and juvenile stages, and magnitude and timing of spawning). We present an analytical method developed by the Geological Survey of Denmark and Greenland (GEUS) in collaboration with the National Institute of Aquatic Resources, Technical University of Denmark (DTU Aqua), for element abundance analysis in otoliths. Analyses of otoliths from Baltic Cod (Gadus morhua; Fig. 1) are used as an example for its application.

Trace element-protein interactions in endolymph from the inner ear of fish: implications for environmental reconstructions using fish otolith chemistry.

Otoliths, the biomineralised hearing "ear stones" from the inner ear of fish, grow throughout the lifespan of an individual, with deposition of alternating calciferous and proteinaceous bands occurring daily. Trace element : calcium ratios within daily increments measured by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) are often used in fisheries science to reconstruct environmental histories. There is, however, considerable uncertainty as to which elements are interacting with either the proteinaceous or calciferous zones of the otolith, and thus their utility as indicators of environmental change. To answer this, we used size exclusion chromatography-inductively coupled plasma-mass spectrometry (SEC-ICP-MS) of endolymph, the otolith growth medium, to determine the binding interactions for a range of elements. In addition, we used solution ICP-MS to quantify element concentrations in paired otolith and endolymph samples and determined relative enrichment factors for each. We found 12 elements that are present only in the proteinaceous fraction, 6 that are present only in the salt fraction, and 4 that are present in both. These findings have important implications for the reconstruction of environmental histories based on changes in otolith elemental composition: (1) elements occurring only in the salt fraction are most likely to reflect changes in the physico-chemical environment experienced during life; (2) elements occurring only in the proteinaceous fraction are more likely to reflect physiological rather than environmental events; and (3) elements occurring in both the salt and proteinaceous fractions are likely to be informative about both endogenous and exogenous processes, potentially reducing their utility in environmental reconstructions.

A rotation-invariant feature space according to environmental applications needs in a data mining system using fish otoliths

Otoliths are microcalcifications found in the inner ear of fishes and their shape can be analyzed to determine sex, age, populations and species, and thus they can provide necessary and relevant information for ecological and environmental studies. This paper compares two different approaches to de al with an arbitrary rotation of a query image in a data mining system that classifies fish otoliths from their images according to the needs of a real environmental application. The different approaches proposed in this paper will allow the successful use of images that are not normalized or are normalized in a different positioning than the images in the database, something that could be very useful in real field applications. The first approach tested is based on a Rotation-invariant Feature Space derived from the Elliptical Fourier Descriptors (EFD), and the second approach relies on a rotation estimation module; in both cases the data mining system is based in a multiclass classifier implemented in this test case with a simple k-Nearest Neighbours after a complex pre-processing step that provides the Feature extraction.

Not All Inner Ears are the Same: Otolith Matrix Proteins in the Inner Ear of Sub-Adult Cichlid Fish, Oreochromis Mossambicus, Reveal Insights Into the Biomineralization Process.

The fish ear stones (otoliths) consist mainly of calcium carbonate and have lower amounts of a proteinous matrix. This matrix consists of macromolecules, which directly control the biomineralization process. We analyzed the composition of this proteinous matrix by mass spectrometry in a shotgun approach. For this purpose, an enhanced protein purification technique was developed that excludes any potential contamination of proteins from body fluids. Using this method we identified eight proteins in the inner ear of Oreochromis mossambicus. These include the common otolith matrix proteins (OMP-1, otolin-1, neuroserpin, SPARC and otoconin), and three proteins (alpha tectorin, otogelin and transferrin) not previously localized to the otoliths. Moreover, we were able to exclude the occurrence of two matrix proteins (starmaker and pre-cerebellin-like protein) known from other fish species. In further analyses, we show that the absence of the OMP starmaker corresponds to calcitic otoliths and that pre-cerebellin-like protein is not present at any stage during the development of the otoliths of the inner ear. This study shows O. mossambicus does not have all of the known otolith proteins indicating that the matrix proteins in the inner ear of fish are not the same across species. Further functional studies of the novel proteins we identified during otolith development are required.

Directional sound sensitivity in utricular afferents in the toadfish Opsanus tau.

The inner ear of fishes contains three paired otolithic end organs, the saccule, lagena and utricle, which function as biological accelerometers. The saccule is the largest otolith in most fishes and much of our current understanding on auditory function in this diverse group of vertebrates is derived from anatomical and neurophysiological studies on this end organ. In contrast, less is known about how the utricle contributes to auditory functions. In this study, chronically implanted electrodes were used, along with neural telemetry or tethers to record primary afferent responses from the utricular nerve in free-ranging and naturally behaving oyster toadfish Opsanus tau Linnaeus. The hypothesis was that the utricle plays a role in detecting underwater sounds, including conspecific vocalizations, and exhibits directional sensitivity. Utricular afferents responded best to low frequency (80-200 Hz) pure tones and to playbacks of conspecific boatwhistles and grunts (80-180 Hz fundamental frequency), with the majority of the units (∼75%) displaying a clear, directional response, which may allow the utricle to contribute to sound detection and localization during social interactions. Responses were well within the sound intensity levels of toadfish vocalization (approximately 140 SPL dBrms re. 1 µPa with fibers sensitive to thresholds of approximately 120 SPL dBrms re. 1 µPa). Neurons were also stimulated by self-generated body movements such as opercular movements and swimming. This study is the first to investigate underwater sound-evoked response properties of primary afferents from the utricle of an unrestrained/unanesthetized free-swimming teleost fish. These data provide experimental evidence that the utricle has an auditory function, and can contribute to directional hearing to facilitate sound localization.

Otolith atlas of fish of the Sinos River

Otoliths are calcium carbonate structures located in the inner ear of fish; they are responsible for hearing and balance. The inner ear has three pairs of otoliths: the lapilli, the sagittae and the asterisci. The sagittae otoliths are the largest and their format is species-specific. Because of their composition, otoliths can resist to the digestive tract of ichthyophagous species, and they can be used as an important tool for identifying species of fish found in stomach contents. The purpose of this work is to provide a photographic guide of the sagittae otoliths of the main fish species from the Sinos River. This atlas consists of photographs of the sagittae otoliths of 36 species belonging to 15 families distributed in five orders.

Novel Natural Transdermal Otoliths/Collagen/Bacterial Cellulose Patch for Osteoporosis Treatment

In the present work, we report the novel natural transdermal otoliths/collagen/bacterial cellulose patch for osteoporosis treatment. This biomaterial is an osteoinductor, or be, stimulates the bone regeneration, enabling bigger migration of the cells for formation of the bone fabric. Otolith is a typical biomaterial that is composed of calcium carbonate and organic matrix. Otoliths are calcareous concrescences present in the inner ear of fishes. Since they are rich in minerals, they are considered essential to the bone mineralization process on a protein matrix (otolin). The objective in this study was to analyze the regeneration capacity of bone defects treated with otoliths network preparation. Collagen and nano-otoliths influences in bacterial cellulose was analyzed using transmission infrared spectroscopy (FTIR). In vivo analysis shows bone surface tissue with high regularity, higher osteoblast activity, and osteo-reabsorption activities areas. These results indicated that the transdermal permeation of otollith using this patch system was sufficient for the treatment of bone diseases. These findings indicate that our novel transdermal delivery system for otolith/collagen/bacterial cellulose is a promising approach to improve compliance and quality of life of patients in the treatment of bone diseases.

Estudo preliminar da utilização de otólitos de Cynoscion acoupa sobre o processo de neoformação óssea em ratos

INTRODUÇÃO: Otólitos são concrescências calcárias presentes no ouvido interno de peixes. Por serem ricos em minerais considerados essenciais ao processo de mineralização óssea sobre uma matriz proteica (otolina), sugere-se que otólitos poderiam funcionar como biominerais. OBJETIVO: Avaliar a capacidade de regeneração de defeitos ósseos tratados com preparado gelatinoso estéril de otólitos. MATERIAL E MÉTODO: O experimento foi realizado com 20 ratos Wistar, distribuídos aleatoriamente em dois grupos, nos quais se realizou defeito ósseo na tíbia, sendo o grupo 1 (G1) experimental, cujas lojas ósseas foram preenchidas com preparado de otólitos, e o grupo 2 (G2) controle, cujas lojas ósseas não receberam tratamento adicional. Os animais foram sacrificados 14 dias após os procedimentos cirúrgicos; as tíbias foram removidas, fixadas em formalina a 10%, descalcificadas em ácido nítrico a 5% e processadas histologicamente. Antes dos sacrifícios, foram coletadas amostras sanguíneas para avaliação das dosagens séricas de cálcio (Ca) e fosfatase alcalina (Alkp). RESULTADOS: As secções histológicas revelaram que o trabeculado ósseo neoformado mostrou-se mais denso e com atividade osteorreabsortiva periosteal menos conspícua no grupo experimental. Entretanto, não demonstraram diferenças estatisticamente significativas na área de neoformação óssea entre G1 (1,9 ± 0,3 mm²) e G2 (1,5 ± 0,4 mm²) (p = 0,0617). Os parâmetros bioquímicos (Ca e Alkp) apresentaram-se dentro dos limites de normalidade. DISCUSSÃO/CONCLUSÃO: Os dados sugerem que os otólitos podem desempenhar papel adjuvante na dinâmica da regeneração óssea.

Auditory hair cell regeneration in teleost fishes: A review.

Exposure to high levels of sound can cause damage to the auditory hair cells of many organisms, including fishes, resulting in hearing deficits. While such exposure can lead to permanent hearing loss in mammals, other phylogenetically older nonmammalian vertebrates, such as birds, reptiles, amphibians, and fishes, can spontaneously regenerate hair cells in both the vestibular and auditory portions of the inner ear following trauma. The general mechanism of hair cell regeneration is currently being examined in a number of different taxa, but data are limited on hair cell regeneration in the inner ear of fishes. This presentation will summarize the few studies that have examined auditory hair cell regeneration in teleost fishes and compare the results reported for fishes to those reported for other species. While understanding the process of hair cell regeneration in fishes has biomedical relevance, this knowledge could also benefit marine managers attempting to mitigate the effects of anthropogenic noise on fishes. [Work supported by NIH P20 RR16481.]

Histochemical localisation of carbonic anhydrase in the inner ear of developing cichlid fish, Oreochromis mossambicus

Inner ear otolith growth in terms of mineralisation mainly depends on the enzyme carbonic anhydrase (CAH). CAH is located in specialised, mitochondria-rich macular cells (ionocytes), which are involved in the endolymphatic ion exchange, and the enzyme is responsible for the provision of the pH-value necessary for otolithic calcium carbonate deposition. In the present study, for the first time the localisation of histochemically demonstrated CAH was analysed during the early larval development of a teleost, the cichlid fish Oreochromis mossambicus. CAH-reactivity was observed already in stage 7 animals (onset of otocyst development; staging follows Anken et al. [Anken, R., Kappel, T., Slenzka, K., Rahmann, H. The early morphogenetic development of the cichlid fish, Oreochromis mossambicus (Perciformes, Teleostei). Zool. Anz. 231, 1–10, 1993]). Neuroblasts (from which sensory and supporting cells are derived) proved to be CAH-positive. Already at stage 12 (hatch), CAH-positive regions could be attributed to ionocyte containing regions both in the so-called meshwork and patches area of the macula (i.e., clearly before ionocytes can be identified on ultrastructural level or by employing immunocytochemistry). In contrast to the circumstances observed in mammalian species, sensory hair cells stained negative for CAH in the cichlid. With the onset of stage 16 (finray primordia in dorsal fin, yolk-sac being increasingly absorbed), CAH-reactivity was observed in the vestibular nerve. This indicates the onset of myelinisation and thus commencement of operation. The localisation of CAH in the inner ear of fish (especially the differences in comparison to mammals) is discussed on the basis of its role in otolith calcification. Since the vestibular system is a detector of acceleration and thus gravity, also aspects regarding effects of altered gravity on CAH and hence on the mineralisation of otoliths in an adaptive process are addressed.

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.

3-D-orientation with the octavolateralis system

Fish detect and localize a sound source with inner ear receptors and with the mechanosensory lateral line. The inner ear of fish is sensitive to the water displacements caused by sound waves through a direct, inertial response by hair cell epithelia of the ear. Hearing specialists, such as goldfish and herring, have accessory peripheral structures that provide additional sensitivity to the pressure component of a sound wave. While the inner ear of fish responds to the whole body motions caused by sound waves and––in case of hearing specialists––to sound pressure, the lateral line is only sensitive to water motions relative to the surface of the fish and to local pressure gradients. Using lateral line and/or acoustic input, some fish can determine the direction and the distance to a sound source. Most likely they do so by exploiting some of the mechanisms described in this paper. Piscivorous fish may use lateral line input to detect the wakes caused by swimming fish. Even in the absence of light catfish, for instance, can follow a 10 s old, three-dimensional wake left by a prey fish over distances up to 55 prey-body length.

No correlation between multilamellar bodies in the inner ear and further organs of mutant (backstroke, bks) and wildtype zebrafish embryos

The origin of the proteinacious matrix of the inner ear stones (otoliths) of vertebrates has not yet been clarified. Using the backstroke mutant (bks) of the zebrafish Danio rerio, which is characterized by a complete lack of otoliths, we searched for possibly missing or aberrant structural components within the macular epithelia of the inner ears of embryos on the ultrastructural level. Numerous multilamellar bodies (MLBs) were found. The MLBs were, however, not restricted to the inner ears of mutants but were also found in wildtype individuals and in further organs such as brain and liver. MLBs have hitherto never been described from the inner ear of fish and are generally estimated to be rare structures. Their occurrence in fish liver can, however, be induced by using particular chemical substances, which seem to effect adaptive compensatory processes on the cellular level. Such a chemical treatment also affects the ultrastructure of further organelles. Since the occurrence of MLBs in the liver of zebrafish was not accompanied by an alteration of the morphology of other organelles, their occurrence seems not to be due to environmental stress. The findings indicate that the MLBs cannot be correlated with bks-inherent features as well as with missing otolith development/growth. Since the occurrence of MLBs was independent from the developmental stage of a specimen and its overall tissue preservation, it can moreover be excluded that these MLBs merely represent fixation artifacts. Their presence more likely indicates cellular remodelling processes of hitherto unknown significance.

Biological factors controlling the carbon isotope record in fish otoliths: Principles and evidence

Otoliths are calcium carbonate structures in the endolymph of the inner ear of fish used to estimate age of individual fish based on the annual and the daily periodicities of the internal banding structure of the otolith. The otolith is about 10% by weight carbon and changes in carbon isotope disequilibrium in the endolymph over the life of the fish are recorded in the pattern of 13C deposition in the otolith. The record of carbon isotope disequilibrium in the otolith can provide useful information about two aspects of the biology of the fish: position in the food chain, and the changes in metabolic rate over the life of the fish. Measurements of σ 13C distribution in otoliths show an increasing 13C enrichment over the part of the life of the fish when respiration rate can be expected to increase, and a decline in 13C when respiration rate can be expected to be decreasing or stationary. Basic models of 13C fractionation in biological systems predict that 13C enrichment of metabolic CO 2 should occur at higher respiration rates. The similarity between observed and expected 13C distributions in otoliths suggests that the enrichment pattern of 13C in otoliths is an analog of the metabolic rate of fishes, which provides a useful life history record.