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At least 19 records

Sturgeon and paddlefish (Acipenseridae) saggital otoliths are composed of the calcium carbonate polymorphs vaterite and calcite: acipenseridae otoliths are vaterite and calcite

The otoliths of modern fishes are most commonly comprised of the metastable aragonite polymorph of calcium carbonate (CaCO3); however, sturgeons have otoliths reportedly comprised of the least stable of the three most-common polymorphs, vaterite. In this study, we used neutron diffraction to characterize CaCO3 polymorph composition of lake sturgeon and paddlefish otoliths. Based on previous summaries of CaCO3 composition over fish evolutionary history, we hypothesized that sturgeon and paddlefish otoliths would have similar polymorph composition. We found that despite previous reports of sturgeon otoliths being comprised entirely of vaterite, that all otoliths we examined in this study also had a calcite fraction that ranged from 17.9+ 6.0 wt. % to 35.9 + 2.9 wt. %. We also conducted a grinding experiment that demonstrated that calcite fractions were due to biological variation and not an artifact of polymorph transformation during preparation. Our study provides the initial characterization of the polymorph composition of the otoliths of lake sturgeon, and paddlefish and also provides the first-ever report of otoliths of Acipenserids as having a calcite fraction.

60 APPLIED LIFE SCIENCES↗

Significance of otolith calcium carbonate crystal structure diversity to microchemistry studies

Otoliths, calcium carbonate (CaCO 3 ) ear stones of fish, contain a wealth of information about fish life and environmental history yet the CaCO 3 polymorph form the otolith is made of is a critical, but seldom considered, piece of information during otolith analysis. Otolith trace element chemistry data increasingly informs management decisions, but recent work has shown that CaCO 3 polymorphs—aragonite, vaterite, and calcite—can bear on incorporation of trace elements in a non-trivial way. Most fishes are thought to have otoliths of the aragonite CaCO 3 form, but this construct is potentially outdated with many recent literature reports showing otherwise. Our study used previously unpublished neutron diffraction data and reports from published literature to address three objectives: (1) summarize the relative effects of otolith CaCO 3 polymorphism on otolith microchemistry, (2) summarize reports of otolith polymorphs to gain a better understanding of the extent of non-aragonite otoliths among fishes, (3) outline future research needed to align interpretations of microchemistry with our current understanding of otolith polymorph diversity. We found that while aragonite otoliths are the most common, so are exceptions. To illustrate, the ostensibly rare (among species) CaCO 3 form vaterite was reported in at least some otoliths of 40% of the species surveyed. Our work suggests that examination of the CaCO 3 polymorph composition of otoliths should become more common particularly in studies where results will or may be used to inform management decisions. Future research should work to attribute controls on otolith CaCO 3 polymorph expression using a combination of -omics and material characterization methods to enrich the life history and environmental information output from otoliths and increase our understanding of the assumptions made in otolith trace element chemistry studies.

58 GEOSCIENCES↗

The incorporation of environmentally derived 87 Sr / 86 Sr and Sr/Ca in early otolith formation of Chinook salmon

Abstract Effective species management often requires understanding patterns of movement and habitat use. A common approach in identifying where individuals reside relies upon chemical tracers from the environment that are incorporated into an individual's tissues. For fish, isotopes in their otoliths, specifically the portion of their otolith formed during their larval stage, have been used to identify the natal origin. Complicating this work, however, is the fact that during this life stage, there is a shift in the source of isotopes deposited onto the growing otolith from maternally to environmentally derived. The objective of this study was to identify the portion of the otolith representing this transition to environmentally derived isotopes so as to accurately investigate questions of natal origin for a threatened population of fall Chinook salmon ( Oncorhynchus tshawytscha ). We exposed developing larvae to four treatments that differed in terms of their water strontium isotope ratio ( 87 Sr/ 86 Sr) and used change‐point analysis of otolith 87 Sr/ 86 Sr and strontium to calcium ratio (Sr/Ca) to identify the otolith radius corresponding to the transition to environmentally derived isotopes. Our results indicated this transition occurred, on average, at 132 μm ( 87 Sr/ 86 Sr; ±50 μm standard deviation) and 127 μm (Sr/Ca; ±29 μm) from the otolith core, which corresponded to the developmental time between hatching and exogenous feeding. A substantial proportion of our otoliths (i.e., 61%) did not show convergence between otolith and water 87 Sr/ 86 Sr by the end of the 113‐day experiment, which was likely due to the dietary contribution of marine‐based feed. Therefore, we were unable to recommend an otolith radius to target for the purposes of reconstructing natal origin apart from being beyond approximately 130 μm.

Chittaro, Paul↗

Influence of ontogenetic development, temperature, and pCO2 on otolith calcium carbonate polymorph composition in sturgeons

Changes to calcium carbonate (CaCO 3 ) biomineralization in aquatic organisms is among the many predicted effects of climate change. Because otolith (hearing/orientation structures in fish) CaCO 3 precipitation and polymorph composition are controlled by genetic and environmental factors, climate change may be predicted to affect the phenotypic plasticity of otoliths. We examined precipitation of otolith polymorphs (aragonite, vaterite, calcite) during early life history in two species of sturgeon, Lake Sturgeon, (Acipenser fulvescens) and White Sturgeon (A. transmontanus), using quantitative X-ray microdiffraction. Both species showed similar fluctuations in otolith polymorphs with a significant shift in the proportions of vaterite and aragonite in sagittal otoliths coinciding with the transition to fully exogenous feeding. We also examined the effect of the environment on otolith morphology and polymorph composition during early life history in Lake Sturgeon larvae reared in varying temperature (16/22 °C) and pCO 2 (1000/2500 µatm) environments for 5 months. Fish raised in elevated temperature had significantly increased otolith size and precipitation of large single calcite crystals. Interestingly, pCO 2 had no statistically significant effect on size or polymorph composition of otoliths despite blood pH exhibiting a mild alkalosis, which is contrary to what has been observed in several studies on marine fishes. These results suggest climate change may influence otolith polymorph composition during early life history in Lake Sturgeon.

54 ENVIRONMENTAL SCIENCES↗

Quantifying fish otolith mineralogy for trace-element chemistry studies

Otoliths are frequently used to infer environmental conditions or fish life history events based on trace-element concentrations. However, otoliths can be comprised of any one or combination of the three most common polymorphs of calcium carbonate - aragonite, calcite, and vaterite - which can affect the ecological interpretation of otolith trace-element results. Previous studies have reported heterogeneous calcium carbonate compositions between left and right otoliths but did not provide quantitative assessments of polymorph abundances. In this study, neutron diffraction and Raman spectroscopy were used to identify and quantify mineralogical compositions of Chinook salmon Oncorhynchus tshawytscha otolith pairs. We found mineralogical compositions frequently differed between otoliths in a pair and accurate calcium carbonate polymorph identification was rarely possible by visual inspection alone. The prevalence of multiple polymorphs in otoliths is not well-understood, and future research should focus on identifying otolith compositions and investigate how variations in mineralogy affect trace-element incorporation and potentially bias environmental interpretations.

60 APPLIED LIFE SCIENCES↗

Partitioning maternal and exogenous diet contributions to otolith 87 Sr/ 86 Sr in kokanee salmon ( Oncorhynchus nerka )

Strontium isotopes ( 87 Sr/ 86 Sr) recorded in the otoliths of Pacific salmon (Oncorhynchus spp.) are commonly used to identify natal origin. For species that migrate at or soon after emergence, the embryonic region of the otolith provides the only record of provenance. However, maternal contribution of Sr from the yolk can confound the isotopic signature of the natal site. We experimentally quantified maternal and exogenous diet contributions to otolith 87 Sr/ 86 Sr over embryonic development in kokanee salmon (Oncorhynchus nerka). Eggs from two populations in isotopically distinct lakes were incubated and reared in a common water source. Timing of developmental events and proportional contribution from yolk to otolith 87 Sr/ 86 Sr differed significantly between the two populations. We suggest that the magnitude of difference in 87 Sr/ 86 Sr between yolk and water, the relative concentrations of Sr and Ca in these isotopic sources, and population-specific effects on otolith growth and composition contribute to this variation. Furthermore, understanding how these factors affect otolith 87 Sr/ 86 Sr could extend the use of otolith geochemistry for determining provenance to species and populations in which natal site rearing is limited.

59 BASIC BIOLOGICAL SCIENCES↗

Evaluation of micromilling/conventional isotope ratio mass spectrometry and secondary ion mass spectrometry of δ 18 O values in fish otoliths for sclerochronology

Stable oxygen isotope ratios (δ 18 O values) measured in fish otoliths can provide valuable detailed information on fish life history, fish age determination, and ocean thermography. Traditionally, otoliths are sampled by micromilling followed by isotope ratio mass spectrometry (IRMS), but direct analysis by secondary ion mass spectrometry (SIMS) is becoming more common. However, these two methods have not been compared to determine which, if either, is better for fish age validation studies. Hence, the goals were to: (1) determine if the δ 18 O signatures from the two different methods are similar, (2) determine which method is better for fish age validation studies, and (3) examine biogeographic and migration history. Both analytical techniques, micromilling/IRMS and SIMS, were used to measure δ 18 O values in six Pacific cod (Gadus macrocephalus) otoliths. A series of measurements was made from the center of each otolith to its edge to develop a life-history δ 18 O signature for each fish. The sampling resolution of SIMS analyses was 2–3 times greater than that obtained by micromilling/IRMS. We found an offset between SIMS and micromilling/IRMS δ 18 O values, about 0.5‰ on average, with SIMS yielding lower values. However, the δ 18 O patterns from both methods (i.e., the number of δ 18 O maxima) correspond to the estimated age determined by otolith growth-zone counts, validating fish age determination methods. Both techniques resolved δ 18 O life-history signatures and showed patterns consistent with seasonal variation in temperatures and changes due to fish migration. When otoliths are large, micromilling/IRMS can provide adequate resolution for fish age validation. Finally, SIMS is the better option if greater sampling resolution is required, such as when otoliths are small or specimens are longer lived and have compact growth zones.

54 ENVIRONMENTAL SCIENCES↗

Characterization of Incremental Markings in the Sagittal Otolith of the Pacific Sardine ( Sardinops sagax ) Using Different Imaging Modalities

Teleost fish possess calcium carbonate otoliths located in separate chambers (utriculus, sacculus, and lagena) of their membranous labyrinth. This study analyzed the surface topography of the sagittal otolith of the Pacific sardine (Sardinops sagax) and the daily and annual increments in these otoliths. The otolith surface, characterized by laser scanning confocal microscopy for the first time, consisted of a system of prominent ridges and valleys (grooves), but it is unclear whether these structures are functional or represent time-resolving markings reflecting growth periodicity. Within the first-year volume, daily increments, each consisting of an incremental (more mineralized) and a discontinuous (less mineralized) zone, were resolved by optical microscopy and backscattered electron (BSE) imaging in the scanning electron microscope (SEM). Daily growth increments could, however, not be resolved in volumes formed after the first year, presumably because otolith growth markedly slows down and spacing of incremental markings narrows in older fish. Throughout otolith growth, the crystalline network continues across the discontinuous zones. Fluorochrome labeling provides additional information on growth after the first year. Compared with optical and BSE imaging, synchrotron microComputed Tomography of intact otoliths (with 0.69 µm volume elements) was less able to resolve daily increments; X-ray phase contrast reconstructions provided more detail than reconstructions with absorption contrast. Future research directions are proposed.

59 BASIC BIOLOGICAL SCIENCES↗

Texture Analysis of Polycrystalline Vaterite Spherulites from Lake Sturgeon Otoliths

Fish otoliths, or ear bones, are comprised of the CaCO 3 polymorphs (aragonite, calcite and vaterite), which can occur either alone or in combination. The polymorph phase abundance in an otolith depends on, as yet, unexplained genetic and environmental factors. Most fish otoliths are comprised of the densest CaCO 3 polymorph, aragonite. Sturgeon otoliths, on the other hand, contain significant amounts of the rare and the structurally enigmatic polymorph, vaterite. Sturgeon otoliths are frequently comprised of agglomerations of small microcrystalline vaterite spherulites (<300 μm in diameter), that range in shape from nearly perfect spheres to oblate spheroids. These spherulites are similar to the synthetic vaterite microspheres employed in laser trapping applications. Vaterite spherulites from both hatchery-reared (juvenile) and wild (adult) Lake Sturgeon exhibit extreme crystallographic texture as evidenced by X-ray diffraction patterns and their reconstructed pole-figures determined here. The vaterite crystallites making up the spherulites have excellent registry in both the axial and equatorial directions. Whether synthesized or natural, the texture manifested in these spherulites suggests that vaterite nucleates and grows similarly in vivo otolith formation as well as from laboratory synthesis. The uniaxial optical character of the vaterite spherulites, confirmed by these diffraction experiments and combined with their large birefringence, makes them well suited for laser trapping applications.

36 MATERIALS SCIENCE↗

Morphology and composition of Goldeye (Hiodontidae; Hiodon alosoides) otoliths

In this work, we provide up-todate morphological and compositional data on otoliths of the osteoglossomorph Goldeye (Hiodon alosoides). Using computed tomography (CT) X-ray, we documented the location of each of the three pairs of otoliths (lapilli, sagittae, and asterisci) in relation to the swim bladder, which extended forward in close proximity to the sagittae and asterisci. The lappili were the largest otoliths in terms of surface area and volume, but the sagittae were highly modified, appearing spiral in shape when viewed dorsally, with a surface area to volume ratio more than double that of the lapilli. Using scanning electron microscopy, the surface of each otolith was viewable in great detail, and small otoconia (~10.5 μm diameter) were observed on each, but were most numerous on the sagittae. Additionally, on scanning electron micrographs, the sagittae appeared to be bi-lobed, with asymmetrical lobes each oriented in the same general direction. Using neutron and X-ray diffraction methods, we found three polymorphs of calcium carbonate crystals (aragonite, vaterite, and calcite), sometimes all within the same otolith. However, in general, lapilli and sagittae were composed predominately of aragonite whereas asterisci were composed chiefly of vaterite. With these results, we provide information on a unique species, whose inclusion in future studies would benefit our understanding of fish hearing, fish evolution, and fisheries ecology.

59 BASIC BIOLOGICAL SCIENCES↗

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

In this paper we quantified the percent CaCO 3 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).

59 BASIC BIOLOGICAL SCIENCES↗

Illuminating otoliths: New insights for life history of Balistes triggerfishes

Our understanding of fish life-history strategies is informed by key biological processes, such as growth, survival/mortality, recruitment and sexual maturation, used to characterize fish stocks (populations). Characterizing the life-history traits of fish populations requires the application of accurate age estimation for managed species. Grey triggerfish Balistes capriscus and queen triggerfish Balistes vetula are important reef-associated species for commercial and recreational fisheries in the Atlantic Ocean. Both species exhibit a unique reproductive strategy for large-bodied fisheries-targeted reef fishes in that they are nesting benthic spawners and invest substantial energy in defence and care of their benthic nests and fertilized eggs. Until recently, our understanding of the life-history strategies of triggerfishes assumed the main method used to obtain age estimates, increments counted from thin sections of the first dorsal spine, provided an accurate characterization of population age-based parameters. However, results from bomb radiocarbon validation studies on the two Balistes species demonstrated that spines do not provide accurate ages, but sagittal otoliths do. The main goal of the current study was to provide an updated understanding for triggerfish life-history strategies by using otolith-based age estimates to characterize population age structure and growth for grey triggerfish and queen triggerfish from waters of the south-eastern U.S. Atlantic. The current study is the first to report on sex-specific age and growth information for grey triggerfish using the Δ 14 C-validated otolith-based age estimation method and the results indicate that the previous characterization of Balistes species as exhibiting moderately rapid growth and as relatively short-lived, based on spine-derived age estimates, are flawed. Otolith-based ages indicated that grey triggerfish and queen triggerfish are moderately slow-growing and long-lived species, attaining maximum ages of 21 and 40 years, respectively. Management efforts for triggerfishes should evaluate these new insights and incorporate the results of otolith-based age estimation into future population monitoring efforts.

59 BASIC BIOLOGICAL SCIENCES↗

Temperature-dependent oxygen isotope fractionation in otoliths of juvenile Chinook salmon ( Oncorhynchus tshawytscha )

Oxygen thermometry has become a widely used technique for reconstructing thermal history in calcifying organisms but interpretation can be subject to predictive error from vital effects. To better understand these processes in Chinook salmon (Oncorhynchus tshawytscha) we experimentally constructed a temperature-dependent, otolith–water fractionation relationship for oxygen isotopes (δ 18 O) by rearing post-yolk absorptive juvenile fish at nominal temperatures from 6 to 21 °C. Temperature and otolith precipitation rate had significant effects on the otolith–water δ 18 O fractionation, but somatic growth rate did not. The slope of the δ 18 O fractionation equation also differed significantly from that of synthetic aragonite. Furthermore, our results suggest the expression of kinetic effects on δ 18 O fractionation in otoliths of Chinook salmon that are increasingly constrained at higher temperatures by other physical and physiological processes involved in mineral formation. Species-specific δ 18 O fractionation equations have considerable utility to reconstruct temperature history in Pacific salmon, but applications should recognize the potential for inferential uncertainty arising from interacting vital effects.

59 BASIC BIOLOGICAL SCIENCES↗

Chemical clocks: using otolith geochemistry to enhance estimation of age and growth of white hake (Urophycis tenuis)

The white hake (Urophycis tenuis) is a groundfish distributed throughout the Gulf of Maine. Catch advice is based on stock assessments done with age-based population dynamics models; however, otolith aging is challenging because of unclear growth increments. To address this concern, we compared the consistency of aging with counts of visual annuli to that of aging with cycles of elemental concentrations measured by using laser ablation inductively coupled plasma mass spectrometry. We tested the hypothesis that oscillations in both environmental conditions and internal physiology through time influence uptake of elements during otolith mineralization. Concentrations of manganese, in comparison with those of the other investigated trace elements (magnesium, strontium, and barium), had the most promising correlation with visual growth increments (~100% age agreement, ±1 year), offering an additional tool to enhance increment identification. In our examination of 550 otoliths collected during 2007–2021, we found that white hake lived a maximum of 10.3 years and exhibited sexual dimorphism in maximum length and age. By using generated von Bertalanffy growth functions, L(t)=110(1–e–0.113(t+0.45)) for males and L(t)=140(1–e–0.113(t–0.30)) for females (where L(t) is length at time t), size and age at maturity were calculated for males (37.4 cm in total length (TL), 3.3 years) and females (47.4 cm TL, 4.2 years). These results demonstrate that otolith geochemistry can be used to improve the accuracy and precision of the estimation of fish age and maturity, even for challenging species.

59 BASIC BIOLOGICAL SCIENCES↗

Otolith 87 Sr / 86 Sr identifies natal origin, movement and life history of Burbot Lota lota in the Kootenai River following 45 years of impoundment

Abstract Hydropower has long been an effective form of renewable energy, but the development of the resource has negatively impacted fishes throughout the world. Formulating recovery plans to mitigate these effects requires a firm understanding of species‐specific provenance, life history and life stage habitat requirements. However, the importance of this information is often only recognised after a substantial population decline, when obtaining it becomes demographically challenging. We provide a case study for this scenario and present a post hoc approach to gain insight into these critical data gaps for Burbot Lota lota in the upper Kootenai River in Montana and British Columbia. The population once supported a robust tribal and recreational harvest but declined following impoundment and these fisheries have remained closed or curtailed for nearly 20 years. The reasons for the decline are unknown and actions to assist recovery have been constrained by limited information regarding natal origin, life history and habitat use. We addressed this issue using strontium isotope ratios ( 87 Sr/ 86 Sr) in otoliths collected over four decades in Lake Koocanusa, the reservoir on the Kootenai River above Libby Dam. Due to the geologic heterogeneity of the basin and resulting variation in water 87 Sr/ 86 Sr, the assignment probability for Burbot to their region of natal origin approached unity. Otolith 87 Sr/ 86 Sr also indicated the population was comprised of mainstem‐ and tributary‐origin forms and their relative abundance changed over time. Mainstem Burbot was most abundant during the 1970s and 2000s, whereas tributary forms dominated between these periods. We suggest ecosystem changes following impoundment contributed to this shift in life history forms and the overall decline of the population. Our findings underscore the need for additional investigations of these impacts to guide restoration efforts.

Dunnigan, James L.↗

Identifying Whitemouth Croaker (Micropogonias furnieri) Populations along the Rio de Janeiro Coast, Brazil, through Microsatellite and Otolith Analyses

The inshore area of the Southwestern Atlantic between 22 °S and 29 °S (South Brazilian Bight) is a transitional climatic zone, where the tropical and warm temperate provinces mix. In its northern part, i.e., in the coastal waters of Rio de Janeiro, Brazil, local oceanographic conditions, such as upwelling in the north, and great bays with different degrees of anthropogenic influences in the center and south can determine the population structure of several fish stocks. The Whitemouth croaker (Micropogonias furnieri) is one the most heavily exploited fishing resources in this area, but there are still some doubts about its population structure. In this study, through combined analyses using nuclear genetic markers and morphological and geochemical signatures of otoliths, a divergence of individuals between two populations was identified using microsatellites, while a finer spatial structure with three populations (north, center and south, respectively) was found based on otolith shapes and elemental signatures. This regional population structure may have direct implications for rational fisheries management and conservation of the species.

Franco, Taynara Pontes (ORCID:0000000167381096)↗

Vaterite Optical Petrography in Lake Sturgeon Otoliths

Vaterite occurring in fish otoliths exhibits plumose and spherulitic habits, the latter being like those grown from aqueous solutions. Vaterite spherulites are known to have sheaf-like texture and can be up to 100 μm in size. In thin section, the spherulites typically show uniaxial positive interference figures between cross-polarizers using a polarizing light microscope but without conoscopic or Bertrand lenses because the spherulites mimic the effect of these lenses. Plumose vaterite can show both uniaxial (+) and biaxial (+) interference figures with a small 2V, which suggests that crystal structural models for vaterite should have the plane of the carbonate group parallel to the optic axis or acute bisectrix. Finally, vaterite spherulites grown from aqueous solution exhibit similar structure and growth habits.

58 GEOSCIENCES↗

Empirically testing vaterite structural models using neutron diffraction and thermal analysis

Otoliths, calcium carbonate (CaCO 3 ) ear bones, are among the most commonly used age and growth structures of fishes. Most fish otoliths are comprised of the most dense CaCO 3 polymorph, aragonite. Sturgeon otoliths, in contrast, have been characterized as the rare and structurally enigmatic polymorph, vaterite a metastable polymorph of CaCO 3 . Vaterite is an important material ranging from biomedical to personal care applications although its crystal structure is highly debated. We characterized the structure of sturgeon otoliths using thermal analysis and neutron powder diffraction, which is used non-destructively. We confirmed that while sturgeon otoliths are primarily composed of vaterite, they also contain the denser CaCO 3 polymorph, calcite. For the vaterite fraction, neutron diffraction data provide enhanced discrimination of the carbonate group compared to x-ray diffraction data, owing to the different relative neutron scattering lengths, and thus offer the opportunity to uniquely test the more than one dozen crystal structural models that have been proposed for vaterite. Of those, space group P6 5 22 model, a = 7.1443(4)Å , c = 25.350(4)Å , V = 1121.5(2)Å 3 provides the best fit to the neutron powder diffraction data, and allows for a structure refinement using rigid carbonate groups.

60 APPLIED LIFE SCIENCES↗