Bias, precision and validation of ageing 0+ European barbel Barbus barbus (L.) from their otoliths

The European barbel Barbus barbus L. is considered a ‘flag’ species for river conservation and sport fishing, but it is increasingly threatened in its native range of distribution. To provide accurate age estimates during early life for appropriate management and conservation measures, the bias and precision of otolith (daily) micro-increment counts were evaluated and age determinations validated on laboratory-reared embryos and larvae. Out of the three pairs of otoliths, the lapillus and sagitta provided reliable age estimates for free embryos and larvae up to 17 days of (known) age post-fertilisation, with first micro-increment formation occurring five days postfertilisation. On the other hand, micro-increments on asterisci formed only 16–17 days post-fertilisation. There was agreement in microincrement counts based on lapilli and sagittae, but not between interpreters, indicating that despite consistency between the two pairs of otoliths extensive training and experience are required for reliable age interpretation. The ability to estimate the ages of 0+ B. barbus from their otoliths will contribute to a better understanding of growth rates from both hatchery-stocked and native/introduced cohorts.     

Forensic biological pursuits of exotic fish origins: piranha in Hawaii

Synopsis The otoliths of an adult red-bellied piranha, Pygocentrus nattereri, captured from a reservoir in Hawaii were scrutinized to determine the fish's origin and growth history. Sagittal otoliths of the piranha, P. nattereri, contained internal microincrements visible by Scanning Electron Microscopy. The medial cross sectional plane of the sagitta was resolved to provide counts for the most visible micro-increments. An opportune spawning of confiscated adults provided samples for verification of daily increment formation. Daily formation of microincrements was verified from hatched individuals, and confirmed the suitability of otoliths for revealing daily patterns in the age and growth of piranhas. The central area of the sagitta was diffuse in regards to otolith microstructure and indicated the fish was held in an unchanging environment (aquarium). Therefore, otoliths provide important life history or forensic information incorporated within their structural components. The visualization of daily microincrements in the otolith of a juvenile allowed the determination of age at and since release. Fish grew rapidly after being released into the wild. From otolith increments the date of release for an individual fish can be calculated with acceptable accuracy. As presented, otolith structural information can provide age and growth data which are essential to the management of introduced species.     

Age estimation and validation for South Pacific albacore Thunnus alalunga

Validated estimates of age are presented for albacore Thunnus alalunga, sampled from a large part of the south‐western Pacific Ocean, based on counts of annual opaque growth zones from transverse sections of otoliths. Counts of daily increments were used to estimate the location of the first opaque growth zone, which was completed before the first assumed birthday. The periodicity of opaque zones was estimated by marginal increment analysis and an oxytetracycline mark–recapture experiment. Both validation methods indicated that opaque zones formed over the austral summer and were completed by autumn to winter (April to August). The direct comparison of age estimates obtained from otoliths and dorsal‐fin spines of the same fish indicated bias, which was assumed to be due to poor increment clarity and resorption of early growth zones in spines, resulting in imprecise age estimates. As such, age estimates from otoliths are considered to be more accurate than those from spines for T. alalunga. This is consistent with results for a growing number of tropical and temperate tuna Thunnini species. It is recommend that validated counts of annual growth zones from sectioned otoliths is used as the preferred method for estimating age‐based parameters for assessment and management advice for these important stocks.     

Rhythmic structural and chemical patterns in otoliths of the Antarctic fish Notothenia larseni: Their application to age determination

Summary Sagittal otoliths from Notothenia larseni contain microincremental growth rings which are distinctly visible in otolith sections using Scanning Electron Microscopy. These microincrements are similar to those deposited daily in the otoliths of fishes from temperate and tropical waters. Microincrements were easily enumerated and fish length was related to increment number by a logarithmic curve. Otolith microstructure analysis appears to provide a technique to accurately determine age and growth rates in these fish. Sr/Ca ratios in otolith aragonite were analyzed along a radius from the outside edge to the core of an otolith section using an electron microprobe. The strontium/calcium (Sr/Ca) ratios varied with a cyclic periodicity apparently related to seasonal water temperature cycles and the number of cycles agreed closely with age estimated from daily microincrement counts. Sr/ca cycles can potentially be used to determine age, validate growth rates determined by other methods, and establish thermal conditions experienced during the life of a fish. Microstructural and chemical analyses of otoliths demonstrate great potential in helping to answer many questions about the growth processes and ecology of Antarctic fishes.     

Age validation,and comparison of otolith,vertebra and opercular bone for estimating age of <Emphasis Type="BoldItalic">Schizothorax o’connori</Emphasis> in the Yarlung Tsangpo River,Tibet

A collection of 514 Schizothorax o’connori was made between August 2008 and August 2009 from Yarlung Tsangpo River to assess the suitability of three bony structures for age estimation. The annulus characteristics of otolith, vertebra and opercular bone were described. Location of the first annulus was validated by daily growth increment (DGI) analysis in the otoliths. Annual periodicity was verified by marginal increment ratio (MIR) analysis in otoliths and edge analysis in vertebrae and opercular bones. Annuli formed, once a year, between March and May for all three bony structures. Otoliths, vertebrae and opercular bones were examined to determine which structure produced the most precise and accurate age estimates in S. o’connori. Vertebrae and otoliths matched closely for the first 21 years of life, while opercular bones appeared to underestimate age. For older fish, the counts diverged and otoliths consistently providing higher age estimates. Sectioned otoliths proved to be the most precise and accurate structure for age estimation. The oldest observed schizothoracine fish was 50, more than twice the longevity previously accepted in S. o’connori.     

Somatic growth and otolith growth in juveniles of a small subtropical flatfish, the fringed flounder, Etropus crossotus

A growth experiment was conducted with juvenile fringed flounder (Etropus crossotus) from North Inlet (South Carolina, USA) to provide information on the growth of a small, shortlived flatfish with a subtropical and tropical distribution. The fringed flounder has a maximum life span of 1.5 y and its long spawning period from March through October complicates the determination of growth rates based on length frequency data. Otoliths of juveniles with a standard length (SL) 23.1-53.0 mm were marked with Alizarin complexone and the fish were held in the laboratory for 66 days at 14, 20, 24 and 29 degrees C while being fed ad libitum. The mean somatic growth increased with temperature from 0.1 mm SL day(-1) at 14 degrees C to 0.4 mm SL day(-1) at both 24 and 29 degrees C. The maximum observed somatic growth rate was 0.7 mm SL day(1) at 29 degrees C. The number of micro-increments formed in otoliths was not significantly different from the expected value, validating formation of one increment per day. The significant relationship between increment width and somatic growth rate can be used to estimate somatic growth rates of individual wild fish based on daily increment information in their otoliths.     

Preliminary notes on the formation of daily increments in otoliths of Oreochromis aureus

Daily growth increments were studied in otoliths of early stage Oreochromis aureus (Cichlidae, Teleostei). A laboratory experiment was carried out on the effect of temperature and food ratio on the otolith growth of juvenile fish. Juvenile O. aureus were reared at two different temperatures, 17°C and 28°C respectively. The young fish were fed two different ratios Trouvit beginning with the first day of swimming and external feeding. Samples were taken at random from each group and the sagitta otoliths were examined. Otolith growth was linearly related to somatic growth of individual fish. Otolith microstructure analysis showed that increment formation began two to three days prior to the transition to the free-swimming stage and continued thereafter following a daily pattern. Temperature and food ratios had a direct influence on the increment widths of the otouths.     

Current methodological refinements for the acquisition of life history information in fishes: paradigms from pan-oceanic billfishes.

1. Many factors contribute to hinder progress in estimating and verifying age, growth, and life history strategies of billfishes. It is necessary to discern growth of an individual fish to circumvent problems in substantiating age, growth, longevity and age at maturity. These biological characteristics allow us to discover what life history periods are critical to survival and reproduction. 2. Two theoretical approaches may be considered for studying the age of billfish: Age may be delineated in terms of the passage of time (chronological age), or may be examined as the progression of physiological development (physiological age). 3. Hard tissues such as otoliths, spines, vertebrae and post-mitotic tissues contain a plethora of biological and ecological information documenting a fish's chronological and physiological age and individual life history. In calcified tissues, this information can be obtained when the proper techniques are used to investigate the structural and chemical components of billfish hard parts. Chemical analyses of stable isotopes and elemental concentrations in calcified tissues suggest that calcified tissues contain hydrographic, nutritional and migrational information--life history information. 4. Utilization of structural and chemical analyses of billfish otoliths, vertebrae, and other hard parts, enables specific phases of growth to be linked to environmental occurrences. Measurement of the metabolically accumulated cellular pigment, lipofuscin, in post-mitotic tissue may make it possible to define physiological age or readiness. Internal and external structural and chemical analyses of otoliths, vertebrae, and other hard parts, combined with biochemical assays of post-mitotic tissue will greatly enhance studies of life history. The methods presented provide insights into means by which life history knowledge can be obtained.     

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.     

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.     

Mediterranean juvenile bluefin tuna: life history patterns

As there is a lack of information on the growth and migrations of bluefin tuna, information about them was gathered using the structural and chemical characteristics of their otoliths and mercury levels in body tissues as indicators of physiological and habitat characteristics. The otoliths of juvenile tuna caught in the Spanish Mediterranean littoral were studied. Otolith increments, assumed to be formed daily, were enumerated. Measurements by wavelength dispersive electron microprobe confirmed the presence of strontium in otolith tissue, and an inverse relationship between strontium/calcium (Sr/Ca) concentration ratio and temperature is suggested. Electron microprobe analyses combined with daily increment analyses of otoliths provided life history profiles for individual fish. Additional Sr/Ca concentration ratio data on fish supported the idea that Sr/Ca ratios can provide information on the environmental history of individual fish. Body concentrations of mercury were related to otolith analyses to suggest age structure, critical life history periods, growth environment, stock structure, food web position, and migration history. The techniques applied present an innovative approach to management-related problems, and the combination of chemical analyses with structural analyses promises to expand our knowledge of the life history of migratory fishes.     

Age and growth of manybar goatfish Parupeneus multifasciatus (Mullidae) from the Nha Trang Bay of the South China Sea

The growth of individuals is analyzed in the mass tropical species manybar goatfish Parupeneus multifasciatus from the Nha Trang Bay of the South China Sea. The relationship between the body weight and body length is determined for the females and males. Among the fishes larger 180 mm fork length (FL), the males prevail. Linear growth of the sagitta is characterized by negative allometry in relation to body length, and sagitta growth in length is faster than in width. Age of fish assessed based on the analysis of daily increments on the sagitta does not exceed three years, and the majority of individuals reach the first sexual maturity by the age of one year. The growth rate of the fish is characterized by a large individual variation.     

Otolith and somatic growth rates in Atlantic salmon parr, Salmo salar L: evidence against coupling

It is often assumed that otolith growth is in some way dependent on somatic growth (i.e. that the two processes are coupled). We examined the relationships between sagitta radius and fork length in 0+ Atlantic salmon parr that would subsequently smolt aged 1 + (UMG fish) or 2+ (LMG fish). Repeated measurements of fork lengths of individually marked parr, taken over a 211-day period from first feeding, were compared to sagitta radii on the same measuring dates (obtained by analysis of daily increments). The results showed that there was a linear relationship between fork length and otolith radius in UMG parr. However, this was not true for LMG parr. These fish enter a state of natural anorexia in their first autumn (despite excess food), but their otoliths continued to grow at the same rate despite the virtual cessation of somatic growth; they had therefore developed disproportionately large otoliths by the end of the study period. The relative growth rates of soma and otoliths first changed in LMG fish in late July/early August; this is the most precise estimate yet obtained of the timing of divergence in the developmental pathways of UMG and LMG parr. The rate of sagitta accretion was consistently lower in LMG parr, possibly indicating a lower metabolic rate in these fish. The results are discussed in relation to previous theories of the relationship between otolith and somatic growth.     

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.     

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.     

Ageing studies done 50 years apart for an inshore fish species from southern Australia—contribution towards determining current stock status

The status of the stocks of Southern Garfish (Hyporhamphus melanochir) in South Australia is currently uncertain. Knowing the extent to which populations have become truncated from fishing activity would help resolve this. Estimates of population structure are available for this species from market sampling done across the two-year periods of 1954 and 1955 and 2006 and 2007. During the latter, fish age was estimated from transverse sections of otoliths using a modern quality assessment and quality control (QA/QC) protocol. The resulting age structures were dominated by the 1+ and 2+ age classes, with older fish contributing less than 16% of harvested numbers. Also, there were few fish >30 cm CFL in size. In comparison, in 1954 and 1955, fish age was determined from whole otoliths, whilst QA/QC procedures were poorly developed. The resulting age structures were dominated by 3+ and 4+ age classes, whilst >25% of the numbers were >30 cm CFL in size. Before concluding that populations were now truncated, it was necessary to reconcile the difference in ageing methods between studies. This was done by assessing the tractability of obtaining accurate estimates of fish age from whole otoliths. Otoliths from a broad age range collected in 2006 and 2007, when immersed in water and displaying the distal face, displayed interpretable patterns of increments. Furthermore, the growth of these otoliths in length slowed but had not stopped even for relatively old fish, indicating that the anterior edge remained a growing edge. A comparison of counts between whole and sectioned otoliths from the same fish gave comparable estimates of age. The studies indicated the tractability of obtaining accurate estimates of age from whole otoliths, when using modern laboratory equipment. However, since cruder techniques were used in the 1950s the accuracy of those ages remained in question. Fortuitously, a small set of otoliths collected in 1955 were available to this study. These were sectioned, counted and compared to counts of the whole otoliths done in the 1950s. Again the counts were comparable, thus conferring confidence about age estimates from the 1950s. The populations of Southern Garfish in 2006 and 2007 were considerably truncated relative to the 1950s, as a consequence of heavy commercial exploitation over the past 50 years. Historical datasets on population structure have made a significant contribution towards understanding current fishery status.     

Sexual dimorphism in the otolith shape of shi drum,Umbrina cirrosa (L.), in the eastern Mediterranean Sea: Fish size–otolith size relationships

Little is known about possible differences in sagitta otolith size and shape between sexes of the shi drum, Umbrina cirrosa, and relationships between their body and otolith size. Thus, this study aimed to fill this knowledge gap via examination of 414 sagittal otoliths from 108 male (total length 13.8–26.8 cm) and 99 female (13.5–26.7 cm) U. cirrosa caught between May 2017 and April 2018 in gillnets set at a depth of ~15 m in Mersin Bay, Eastern Mediterranean Sea. No statistical differences were observed between the shape indices of the left-sided and right-sided sagitta. However, there were significant differences in the size and shape of otoliths between males and females. The slopes of allometric power functions from otolith width × fish sizes gave significant differences between males and females (ANCOVA, P < 0.05). The relationship for length × weight of otoliths from both males and females showed isometric growth, whereas the relationship of otolith width × otolith weight showed positive allometry. Negative allometric growth was observed for the relationship otolith length × otolith width. In summary, this study revealed the presence of sexual dimorphism in the otolith shape of U. cirrosa, and the data on regression relationships of fish-otolith sizes can be used to estimate fish size from U. cirrosa otolith sizes.     

Vaterite Otoliths from the Opah,Lampris immaculatus,and Two Species of Sunfish,Mola mola and M. ramsayi

The cosmopolitan opah Lampris immaculatus (also known as the moonfish or mariposa) occurs commonly in New Zealand waters. The otoliths of 23 individuals were examined by light microscopy. All three of the otoliths of the endolymphatic sac of the opah have the characteristic appearance of the Stolkowski crystal form of the vaterite morph of calcium carbonate. Five randomly chosen pairs of asterici and sagittae and two lapilli otoliths were shown by X-ray diffraction to be composed of vaterite. In addition to their mineral composition, the sagitta and astericus of the opah present an unusual combination of anatomical features found in both what are generally regarded as primitive and as advanced fish. Vaterite otoliths also occur in the sunfish Mola mola and M. ramsayi, but with a different crystalline habit to that found in the opah, and in a distinctively different anatomical arrangement of the endolymphatic sac. It is argued that, based on their otolith mineralogy and morphology, the opah and the sunfish are each side of one of the biochemical and anatomical boundaries involved in the separation of the teleosts from other actinopterygian fish (sturgeons, paddle fish, gars and bowfins).     

Evaluating the effectiveness of teeth and dorsal fin spines for non‐lethal age estimation of a tropical reef fish,coral trout Plectropomus leopardus

This study investigated whether teeth and dorsal fin spines could be used as non‐lethal methods of age estimation for a vulnerable and highly valued tropical fisheries species, coral trout Plectropomus leopardus. Age estimation of individuals from 2 to 9 years old revealed that dorsal spines represent an accurate ageing method (90% agreement with otoliths) that was more precise [average per cent error (APE) = 4·1, coefficient of variation (c.v .) = 5·8%] than otoliths (APE = 6·2, c.v . = 8·7%). Of the three methods for age estimation (otoliths, dorsal spines and teeth), spines were the most time and cost efficient. An aquarium‐based study also found that removing a dorsal spine or tooth did not affect survivorship or growth of P. leopardus. No annuli were visible in teeth despite taking transverse and longitudinal sections throughout the tooth and trialling several different laboratory methods. Although teeth may not be suitable for estimating age of P. leopardus, dorsal spines appear to be an acceptably accurate, precise and efficient method for non‐lethal ageing of individuals from 2 to 9 years old in this tropical species.     

Eco-morphological patterns of the sagitta of Antarctic fish

Morphology and morphometry of the sagittae otolith were studied in pelagic and mesopelagic fish. The shape, margins and rostrum of four groups of otoliths from several species were analyzed: group 1 (pelagic fish associated with the under ice cover N = 42), group 2 (pelagic fish associated with water offshore N = 9), group 3 (mesopelagic fish associated with extensive vertical migration N = 57) and group 4 (mesopelagic fish associated with short vertical migration N = 54). E (maximum width of the sagitta /maximum length of the sagitta %), R (rostrum length (RL)/maximum length of the sagitta %) and S (sulcus area (SS)/otolith area (OS) %) indexes were calculated for each species. Sagittae of pelagic groups (1 and 2) showed the smallest sagitta dimensions in relation to the total length of the fish, in this group the sagitta shape is variable. Sagittae of mesopelagic fish (groups 3 and 4) showed variable shape and edges. The shape in group 4 was polygonal and these species have more width than length. Statistical analysis showed significant differences in the E, R and S indexes. These results were compared with other 19 species, belonging to six families, taken from a publisher-edited literature. E, R and S-values could be used to characterize the sagittae of the Antarctic fish and could be considered as a useful tool for fish ecology studies.