Otolith shape analysis and daily increment validation during ontogeny of larval and juvenile European chub Squalius cephalus

This assesses features of otoliths from laboratory-reared embryos, larvae and juvenile European chub Squalius cephalus from hatching to 180 days post-hatching (dph). We observed the development of the three pairs of otoliths (lapilli, sagittae and asterisci) and more precisely shape changes, as well as timing and deposition rate of increments of the lapilli. The lapilli and the sagittae were present at hatching, whereas the asterisci formed between 20 and 30 dph. The lapillus and sagitta shapes were round until 20 dph. From 60 dph the anterior and the posterior rostra of the sagittae were well developed, but very thin, making this otolith too fragile to manipulate for further studies of shape and validation of otolith increment deposition rate. The lapilli provided reliable age estimates for free embryos, larvae and juveniles up to 120 dph. However, caution should be taken when ageing fish older than 150 dph as an underestimation was noticeable. The regression of the number of otolith increments on age showed a slope and an intercept not significantly different from 1 and 0, respectively, which indicated that otolith growth increments were deposited on a daily basis, with the first microincrement occurring at hatching. Increment counts were consistent between three interpreters, indicating a consistent and reliable age estimate. This study validates that the otolith increment deposition rate can be used to assess hatching dates and daily growth of wild S. cephalus under 150 dph and in environments similar to the conditions used in this study.     

An evaluation of the otolith characteristics of Conger conger during metamorphosis

A sample of 20 metamorphosing conger eel Conger conger leptocephali were collected from the Minho River, Portugal, in February 1999 and their sagittal otoliths were analysed by scanning electron microscopy. Four different etching agents were applied along both sagittal and frontal sections during otolith preparation to examine the microstructural growth in this species. Otolith growth increments were visible throughout the increment countable zone using all four treatments, but a permanent peripheral diffuse zone, where the daily increments were unclear, appeared on all otoliths, preventing accurate age estimation. To understand more about the nature of the diffuse zone, otoliths of 10 other metamorphosing leptocephali reared in aquaria were marked by immersion in tetracycline hydrochloride. The distance between the fluorescent marks and otolith edge, measured over a fixed period of time, was used to estimate the otolith growth rate. The application of this technique led to an anomalously high estimated otolith growth rate, probably as a result of the capture, marking and handling stress.     

Otolith Microstructure of Larval Gymnocypris potanini Herzenstein from the Minjiang River in China

Synopsis Otolith microstructure of about 120 Gymnocypris potanini larvae from the Minjiang River in China was examined and analyzed. Larvae had multiple primordia in most lapilli and sagittae, while had only one primordium in a few specimens. There had only one nucleus in otoliths of the larvae, except for some few specimens with 2 nuclei. The transparence of many otoliths differed from center to edge, and part of them could be divided into inner low optically dense zone (LODZ) and outer optically dense zone (ODZ). Based on increment clarity, otoliths of this species could be classified into three types, which were otolith with subtile increments, otolith with almost identified increments, and otolith with fairly clear increments characterized by high contrast. The last two types of otolith accounted for 87.07% in lapilli and 94.46% in sagittae, respectively. Increment clarity of sagitta was higher than that of lapillus. Natural checks were identified in 32.50% lapilli and 48.33% sagittae. These checks primarily located in the first to sixth increment. According to the number of increments in otoliths, the age of this batch larvae was 14 – 22 days, birth date was on June 17 – 25, and average growth rate of body length was 0.8936 ± 0.08769 mm/d.     

Validation of a technique for reconstructing daily patterns in the recruitment of coral reef damselfish

Counting growth increments in otoliths recently has become an accepted method of ageing tropical fishes, however, verification is essential for each new species. In this study, growth increments in otoliths of the juveniles of several coral reef damselfishes (Pomacentridae) were deposited daily and a distinct transition from wide to narrow increments coincided with settlement from the pelagic larval phase into the demersal habitat. Thus, the data of settlement for each individual fish could be calculated with acceptable accuracy. The daily pattern of recruitment to a coral reef was successfully reconstructed using the otoliths from a large sample of juvenile fish collected at the end of the breeding season. This was because the original pattern of settlement was preserved in the age distribution for at least 4 to 5 months. This application of otolith ageing techniques may be extended to reveal the temporal patterns of recruitment to many localities encompassing spatial scales larger than would be logistically possible using visual censuses.     

Otolith formation,microstructure and daily increment validation in juvenile perch Perca fluviatilis

The otoliths of laboratory‐reared larval and juvenile perch Perca fluviatilis of known age were analysed to determine the age of otolith formation and validate the formation of daily increments. There was a linear relationship between number of increments and age in days post‐hatching, although by 82 days post‐hatching daily increment counts underestimated actual age by an average of 5 days. Otolith dimensions in relation to standard length indicated allometric growth of otoliths until completion of yolk absorption, and isometric growth thereafter, up to 82 days post‐hatching.     

The use of sectioned otoliths to age barramundi (Lates calcarifer) (Bloch, 1790) [Centropomidae]

The relationship between the number of rings present in sagittal otoliths and the age of barramundi, Lates calcarifer (Bloch, 1790) [Centropomidae], was investigated by examining cross sectioned otoliths of 37 tagged fish of known age between 1 and 5 years from the Johnstone River, north Queensland. Concentric rings were clearly visible in all otolith sections and were validated as annual marks. The technique was then used to estimate the age and calculate von Bertalanffy growth parameters for 70 barramundi from the Fitzroy River, central Queensland. Growth appeared to be rapid but variable in the first year; the von Bertalanffy growth parameters for length versus age were L =690 mm, K=0.53, t ₀= 0.003 years. October 1 was designated as the birth date. Whole otolith length, width and thickness were also approximated well by the von Bertalanffy equation. We suggest that examination of otoliths is a useful technique for ageing barramundi but note that further validation of the ageing method is still needed for fish older than six years.     

Correlations between body length and otolith size in smallmouth bass Micropterus dolomieu Lacépède, 1802 with implications for retrospective growth analyses

Reconstructing individual growth history from analysis of increments in otoliths, scales, or spines can provide information on past growth responses to environmental variation, which in turn can be useful for predicting population‐level response to climate change. The objective of this study was to examine correlations between body length and different metrics of otolith size for Micropterus dolomieu. Three metrics corresponding to commonly‐used microstructural and ultrastructural otolith dimensions were measured using image analysis of digital micrographs from a sample of 214 M. dolomieu ranging from 115 to 438 mm total length collected in 2011–2013. It was found that anteroposterior length of whole otoliths provided much improved regression relationships with body size as well as ease of data collection and faster sampling throughput compared with microstructural measures from polished sections. When applying these metrics to reconstruct growth history the biological intercept model generally produced more reasonable back‐calculated estimates of length‐at‐age, although this was not consistent across all otolith metrics. Results suggest that whole otolith measures should be employed due to efficiency of data collection and greater reliability for reconstructing growth history in M. dolomieu.     

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.     

Validation of daily otolith increments in larval and juvenile Chinese sucker, <Emphasis Type="Italic">Myxocyprinus asiaticus</Emphasis>

Otolith development was observed and the formation of daily growth increments in otoliths of Chinese sucker, Myxocyprinus asiaticus, was validated by monitoring known-age larvae and juveniles in the laboratory from 2003 to 2005. Otolith shape changed with larval and juvenile development, and there was an exponential relationship until a body length of 16 mm or so, and a linear relationship after a body length of 16 mm between otolith size and fish size. The first increment was identified in larvae 1 day after hatching. The regressed equations between daily age (D) and increment number in otoliths (N) were N = −0.64 + 0.96D in lapillus, and N = −0.31 + 0.98D in sagitta. The slopes were not significantly different than 1.0. This demonstrated that otolith increments in this species were formed daily and can be used for daily age determination.     

Comparison of growth rate and formation of otolith increments in age-0 red snapper

For wild red snapper Lutjanus campechanus , mean otolith increment deposition rate after marking with oxytetracycline dihydrate (OTC) was daily (0.97 increments day⁻¹) when growth rates were fast (0.63 mm fork length, L _F day⁻¹), but were not daily (0.82 increments day⁻¹) when somatic growth was slow (0.2 mm L _F day⁻¹). For reared larvae ( n =8), increment deposition rates were daily (0.99–1.03 increments day⁻¹), and growth rates ranged from 0.6 to 0.9 mm L _F day⁻¹. Growth rate affected increment deposition rate as a threshold function, i.e. when growth rate was <0.3 mm L _F day⁻¹, deposition was less than daily, but above this level increment deposition did not exceed a daily rate. As growth rates increased increment widths increased. Examination of a sub-sample ( n =8) of the otoliths from the slowest growing wild fish by scanning electron microscopy did not increase increment counts. Because L. campechanus are late spring-early summer spawners, young fish can expect maximum growth due to warm summer temperatures. Thus, daily ageing methods should be well suited to this species.     

Do otolith increments allow correct inferences about age and growth of coral reef fishes?

Otolith increment structure is widely used to estimate age and growth of marine fishes. Here, I test the accuracy of the long-term otolith increment analysis of the lemon damselfish Pomacentrus moluccensis to describe age and growth characteristics. I compare the number of putative annual otolith increments (as a proxy for actual age) and widths of these increments (as proxies for somatic growth) with actual tagged fish-length data, based on a 6-year dataset, the longest time course for a coral reef fish. Estimated age from otoliths corresponded closely with actual age in all cases, confirming annual increment formation. However, otolith increment widths were poor proxies for actual growth in length [linear regression r ² = 0.44–0.90, n = 6 fish] and were clearly of limited value in estimating annual growth. Up to 60 % of the annual growth variation was missed using otolith increments, suggesting the long-term back calculations of otolith growth characteristics of reef fish populations should be interpreted with caution.     

Preliminary estimates of the age and growth of immature smooth oreo Pseudocyttus maculatus Gilchrist 1906 (Oreosomatidae) in the Falkland Islands region of the South Atlantic

Sectioned otoliths were used to estimate the age of 29 specimens of the smooth oreo (Pseudocyttus maculatus Gilchrist 1906) from the Falkland Islands region of the South Atlantic. This represents the first ageing study of this species in this region of the world. All specimens were immature (sub-adults), the largest having a maximum total length of 347 mm. Growth increments were observed in sectioned otoliths and assumed to be annual in periodicity. The maximum age estimate of individuals in this study was 20 years and the form of somatic growth was similar to previous research on this species from Australian waters. The otolith microstructure consisted of an inner, mostly opaque zone with wide increments followed by an outer, mostly translucent zone with narrower increments. It is suggested that these zones correspond to the two growth phases in the life history of this species, namely the pelagic juvenile phase (opaque zone) and the demersal sub-adult/adult phase (translucent zone). Received: 27 May 1997 / Accepted: 31 October 1997     

Daily growth increments in otoliths of milkfish, Chanos chanos (Forsskål), larvae

The formation of daily growth increments of otoliths was studied in the reared larvae of milkfish. The first growth increment was formed during the yolk–sac reabsorption period c .2 days after hatching, and increment formation continued on a daily schedule regardless of growth rate. The initial incremental zone was of amorphous structure, and the subsequent incremental zone was of needle–shaped crystalline structure; the former structure was formed in the yolk–sac reabsorption period of the larva and the latter in the exogenous feeding period. Accordingly, ageing of milkfish larvae is possible by counting growth increments, and timings within the developmental stage of the larvae can be understood by examining the otolith microstructure.     

Factors affecting morphological development of the sagittal otolith in juvenile and adult small yellow croaker (Larimichthys polyactis Bleeker, 1877)

The morphology and morphometrics of the sagittal otoliths of small yellow croaker (Larimichthys polyactis) from the southern Yellow Sea were investigated. Study objectives were to evaluate the shape variability and morphometric variables of sagittae of juveniles and adults as related to developmental changes and habit shift. A total of 152 fish were sampled from April to June of 2012 and 2013, along the coastal waters of the Lüsi spawning ground in the southern Yellow Sea. Changes in otolith shapes from the juvenile to the adult were presented with the rim development through the entire‐lobed‐entire transition and with the curvature of the cauda toward the ventral margin. The otolith elongation in the juvenile stage occurred at 10–20 mm standard length (SL) and was likely associated with the formation of otolith accessory growth centers from larvae to juvenile. The L. polyactis sagittal otoliths acquired their definitive shape at 130 mm SL maturity. Ontogeny on otolith shape might be related, for example, to the factors of metamorphic development, feeding habitat, and ambient water salinity, which varied throughout the growth of L. polyactis.     

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.     

Three-dimensional imaging of walleye pollock otoliths: reconstruction from serial sections and fluorescent laser cytometry

Two techniques have been developed to examine the three-dimensional internal structure of otoliths. In the first, otoliths were sectioned serially, images were digitized, and the otolith was reconstructed as a computer model. In the second method growth increments were marked in vivo during their formation by immersing the fish in a fluorescent dye, and then the internal structure of the otolith visualized using laser cytometry. The results are useful for evaluating the potential for bias in otolith measurements and for determining the sectional plane with the least bias.     

Comparison of otolith growth and morphology with somatic growth and age in young-of-the-year bluefin tuna

Otolith morphological characteristics were studied using image analysis techniques and the relationships between otolith growth and somatic growth and age, as estimated from counting daily otolith increments, were examined in young-of-the-year (YOY) bluefin tuna Thunnus thynnus ranging in fork length ( L _F) from 8·5 to 55·5 cm. Whole otolith length, width, area and perimeter, and three shape indexes, circularity, E value and rectangularity, were extracted for each pair of sagittae. Since no statistical significant differences between left and right otolith morphometrics were found, only one otolith from each fish was used for correlations. Statistically significant relationships were observed between otoliths measurements and fish somatic growth when a linear regression was applied after logarithmic transformation of all variables tested. Among the variables, otolith length was the one that showed the highest correlation with L _F, followed by otolith area and perimeter, whereas otolith rectangularity exhibited the lowest correlation. Statistically significant relationships were also observed between the otolith variables tested and the age of the fish, which ranged from 20 to 129 days. The ages estimated using otolith mass were very close to those assessed using daily increment counts (bias ranged from 1 to 24 days). Therefore, otolith mass could represent a valuable criterion for age estimation in YOY bluefin tuna that is objective, economic and easy to perform compared to daily increment counting method.     

Growth and morphological development of sagittal otoliths of larval and early juvenile Trachurus japonicus

The daily periodicity of growth increment formation in sagittal otoliths of jack mackerel Trachurus japonicus was validated by marking otoliths with alizarin complexone (ALC). Analysis of otoliths of known‐age juveniles confirmed that the first increment formed on day 3 after hatching, and was associated with first feeding. A total of 198 specimens, ranging from 2·6 to 49·2 mm in body length (notochord length or standard length) and from 7 to 78 days in age, were collected in the East China Sea and Tosa Bay, and used to examine the association between otolith morphological development and ontogenetic development. The relationship between body length ( L ) and otolith radius ( R ) was significantly described by the linear function L  = 2·65 + 0·0425 R ( n  = 198, r ² = 0·99, P  < 0·001), indicating that somatic growth history can be reconstructed from otolith growth patterns. The otolith was primarily spherical in the preflexion larval stage, and became elongated with notochord flexion. The first secondary primordium formed at c . 25 days, during the middle postflexion stage, and was associated with metamorphosis. By c . 42 days the sagittal otolith was adult‐like in morphology, with the primary growth zone enclosed by the marginal growth zone, except in the anterior rostrum area. Thus age, growth and developmental stages were recorded in sagittal otoliths during the larval and early juvenile stages of jack mackerel.     

Tetracycline tagging in coregonid embryos and larvae

Coregonus peled (Gmelin) embryos at the eyed stage were immersed in tetracycline hydrochloride (TC) solution (600 mg l⁻¹) and fluorescent marks on the otoliths from fish were identified under a UV light microscope. Three weeks after treatment, when the larval fish had hatched, multiple primordia of the sagitta had fluorescent bands whose locations suggest that calcification begins before formation of the otolith's core. Upon hatching, increments were few or absent, and daily increments started forming after hatching. In coregonid larvae immersed in TC solution immediately after hatching, the fluorescent band was only 1 day's growth increment wide in the otolith. The age of larvae based on ring counts after marking corresponded to the actual age, but further studies are required. Triple marks due to 35-h immersions in TC solution separated by 5–7 day feeding periods were clearly readable in the otoliths. One month after treatment the fluorescent marks were identified under UV light on otoliths from every fish. In treated embryos, 38% of fish had marks 3 months after TC immersion, whereas in treated larvae 60–80% of fish grown up to 38.7 mm were marked.     

有明银鱼耳石显微结构和微化学研究

1998年从鸭绿江捕取有明银鱼成鱼 ,生物学测量后剖出耳石。每尾鱼 1对矢耳石中的 1个用于制片 ,显微镜下观察其显微结构 ,另 1个制片镀碳膜后 ,用扫描电子微探针测定Sr和Ca的浓度比率。Sr和Ca分析的标准样用SrSO4 和CaSO4 。 31尾鱼 (体长 116～ 15 7mm)耳石长半径 (Y)与鱼体长 (X)成线性关系 ,其方程式为Y =2 72 16X +9 2 2 92 ,r =0 9178,P <0 0 1。耳石中心和耳石原基的平均直径分别为 2 2 5 5± 2 88μm和6 95± 1 30 μm。 13尾鱼耳石上的日轮数为 30 5～ 35 4,大约在 45～ 70日轮处出现过渡轮。日轮间距 0 94～ 1 14μm ,最大间距在 190～ 2 2 0日轮处 ,最小间距出现在 2 80日轮之后。耳石中Sr、Ca浓度比例在第 5至第 7测点出现第 1个高峰 ,这与稚鱼为了越冬洄游到海中时间相吻合。第 2个高峰出现在 2 8～ 32测点 ,这与鱼产卵上溯洄游时间相符。个体发育过程中生理和生态条件的变化 ,可引起耳石中Sr、Ca比率的改变 ,但栖息水域盐度的突然改变是导致Sr、Ca比率显著增高的主导因素。