Periodicity of the growth-band formation in vertebrae of juvenile scalloped hammerhead shark Sphyrna lewini from the Mexican Pacific Ocean

The age of 296 juvenile scalloped hammerhead sharks Sphyrna lewini caught by several fisheries in the Mexican Pacific Ocean from March 2007 to September 2017 were estimated from growth band counts in thin-sectioned vertebrae. Marginal-increment analysis (MIA) and centrum-edge analysis (CEA) were used to verify the periodicity of formation of the growth bands, whereas elemental profiles obtained from LA-ICP-MS transect scans in vertebrae of 15 juveniles were used as an alternative approach to verify the age of the species for the first time. Age estimates ranged from 0 to 10+ years (42–158.7 cm total length; L_T). The index of average percentage error (I_APE 3.6%), CV (5.2%), bias plots and Bowker's tests of symmetry showed precise and low-biased age estimation. Both MIA and CEA indicated that in the vertebrae of juveniles of S. lewini a single translucent growth band was formed during winter (November–March) and an opaque band during summer (July–September), a period of faster growth, apparently correlated with a higher sea surface temperature. Peaks in vertebral P and Mn content spatially corresponded with the annual banding pattern in most of the samples, displaying 1.19 and 0.88 peaks per opaque band, respectively, which closely matched the annual deposition rate observed in this study. Although the periodicity of growth band formation needs to be verified for all sizes and ages representing the population of the species in the region, this demonstration of the annual formation of the growth bands in the vertebrae of juveniles should lead to a re-estimation of the growth parameters and productivity of the population to ensure that it is harvested at sustainable levels.     

Elemental Markers in Elasmobranchs: Effects of Environmental History and Growth on Vertebral Chemistry

Differences in the chemical composition of calcified skeletal structures (e.g. shells, otoliths) have proven useful for reconstructing the environmental history of many marine species. However, the extent to which ambient environmental conditions can be inferred from the elemental signatures within the vertebrae of elasmobranchs (sharks, skates, rays) has not been evaluated. To assess the relationship between water and vertebral elemental composition, we conducted two laboratory studies using round stingrays, Urobatis halleri, as a model species. First, we examined the effects of temperature (16°, 18°, 24°C) on vertebral elemental incorporation (Li/Ca, Mg/Ca, Mn/Ca, Zn/Ca, Sr/Ca, Ba/Ca). Second, we tested the relationship between water and subsequent vertebral elemental composition by manipulating dissolved barium concentrations (1x, 3x, 6x). We also evaluated the influence of natural variation in growth rate on elemental incorporation for both experiments. Finally, we examined the accuracy of classifying individuals to known environmental histories (temperature and barium treatments) using vertebral elemental composition. Temperature had strong, negative effects on the uptake of magnesium (D_Mg) and barium (D_Ba) and positively influenced manganese (D_Mn) incorporation. Temperature-dependent responses were not observed for lithium and strontium. Vertebral Ba/Ca was positively correlated with ambient Ba/Ca. Partition coefficients (D_Ba) revealed increased discrimination of barium in response to increased dissolved barium concentrations. There were no significant relationships between elemental incorporation and somatic growth or vertebral precipitation rates for any elements except Zn. Relationships between somatic growth rate and D_Zn were, however, inconsistent and inconclusive. Variation in the vertebral elemental signatures of U. halleri reliably distinguished individual rays from each treatment based on temperature (85%) and Ba exposure (96%) history. These results support the assumption that vertebral elemental composition reflects the environmental conditions during deposition and validates the use of vertebral elemental signatures as natural markers in an elasmobranch. Vertebral elemental analysis is a promising tool for the study of elasmobranch population structure, movement, and habitat use.     

Evaluation of copper, iron and lead substitution techniques in elasmobranch age determination

Copper and lead substitution reactions were found to be useful in demonstrating growth patterns in vertebrae from several elasmobranch species. In contrast, iron substitution was ineffective in detecting variations in vertebral mineralization. The present study provides two alternative methods for elasmobranch age determination that may prove useful when traditional methods are deemed ineffective. However, species-specific variations in the efficiency of these methods reemphasize the observation that different types of vertebrae often require different methods of staining.     

Age and growth of the tropical oviparous shark,Chiloscyllium punctatum in Indonesian waters

The brown-banded bamboo shark, Chiloscyllium punctatum, is the most common shark caught in coastal commercial fisheries throughout Southeast Asia, yet there is a lack of the life-history information necessary for reliable stock assessments. The authors estimated growth rates and age at maturity using analysis of growth bands in vertebral centra. They trialled four different techniques to enhance the visibility and improve identification of the putative annual growth bands necessary for age estimation. The authors found that the burn method on whole vertebral centra provided the most readable and consistent results for age analysis. The logistic model was chosen as the best-fit growth model for age estimation of 330 individual C. punctatum from Indonesia. Several age verification methods, including marginal increment ratio and length-frequency analysis, were performed with the support of age validation through the use of calcein-labelled vertebrae from two sharks maintained in captivity. This study found that C. punctatum from Indonesian waters is a fast-growing species that can grow up to 18 cm year⁻¹, reach an estimated maximum total length of 1 m, mature at c. 6.5 years and live for up to 14 years.     

Natural variability and effects of cleaning and storage procedures on vertebral chemistry of the blacktip shark Carcharhinus limbatus

Key methodological assumptions regarding the degree of natural variability and influence of sample handling and storage of elasmobranch vertebral chemistry were assessed using laser‐ablation inductively coupled plasma mass spectrometry. Vertebral chemistry of juvenile blacktip sharks Carcharhinus limbatus was examined to identify whether differences existed among different regions of the vertebral column, between thoracic vertebrae of individual fish or within individual vertebrae. Additionally, the effects of bleach exposure and storage in ethanol on vertebral chemistry were compared. No significant variation in vertebral chemistry was found among different regions of the vertebral column or between thoracic vertebrae, but significant differences among transect locations within individual vertebrae were observed. The variation at all three levels appears comparable with published data on sagittal otoliths of bilaterally symmetrical teleost fishes. The experimental assessment of potential treatment effects indicated vertebral chemistry was not significantly affected by bleach or exposure to ethanol. Taken together, these results support the idea that vertebrae taken from the same region of the vertebral column can be treated as equivalent and at least certain elements remain robust to exposure to bleach and ethanol.     

Assessing the accuracy of using whole and sectioned vertebrae to determine the age of an endemic sisorid catfish, <Emphasis Type="Italic">Glyptosternon maculatum</Emphasis>, in Tibet,China

The purpose of this study was to develop a reliable method of age estimation for Glyptosternon maculatum (Sisoridae, Siluriformes) in the Yarlung Tsangpo River basin of Tibetan Plateau using whole and sectioned vertebrae. Sectioned vertebrae showed that translucent bands were separated by distinct opaque bands through the focus to the margin. Although whole vertebrae showed clear band patterns in the intermediate region, such band patterns are obscure around the region of the focus of the vertebrae in older fish. The whole and sectioned vertebrae methods produced consistent ages until age 6, but there was a trend towards greater differences between the two methods as age increased. These findings indicate that sectioned vertebrae are suitable for age estimation of this species.     

An analysis of the feasibility of using caudal vertebrae for ageing the spinetail devilray, Mobula japanica (Müller and Henle, 1841)

Age assessment of Mobula japanica may be possible using the caudal vertebrae, below the origin of the dorsal fin. This is supported by the significant linear relationship found between disc width (DW) and centrum radius (CR, n?=?55), the continuous record of growth bands in the vertebrae, the clarity to distinguish and count growth bands, and the precision of the band counts. Assuming an annual band formation, the preliminary assessment of the age suggests that M. japanica lives at least 14 years and has a low growth rate (K?=?0.28 year^?1). The minimum number of growth bands was one for spinetail devilrays with a 1,210–1,390 mm DW, while the maximum was 14 for a 2,300 mm DW devilray. While age validation is still required, results indicate the feasibility of the use of caudal vertebrae for age estimation. To provide robust estimates of validated age and growth for the spinetail devilray, the sampling coverage needed might imply an international cooperation.     

Investigations of Δ 14C, δ 13C, and δ 15N in vertebrae of white shark (Carcharodon carcharias) from the eastern North Pacific Ocean

The white shark, Carcharodon carcharias, has a complex life history that is characterized by large scale movements and a highly variable diet. Estimates of age and growth for the white shark from the eastern North Pacific Ocean indicate they have a slow growth rate and a relatively high longevity. Age, growth, and longevity estimates useful for stock assessment and fishery models, however, require some form of validation. By counting vertebral growth band pairs, ages can be estimated, but because not all sharks deposit annual growth bands and many are not easily discernable, it is necessary to validate growth band periodicity with an independent method. Radiocarbon (¹⁴C) age validation uses the discrete ¹⁴C signal produced from thermonuclear testing in the 1950s and 1960s that is retained in skeletal structures as a time-specific marker. Growth band pairs in vertebrae, estimated as annual and spanning the 1930s to 1990s, were analyzed for Δ¹⁴C and stable carbon and nitrogen isotopes (δ¹³C and δ¹⁵N). The aim of this study was to evaluate the utility of ¹⁴C age validation for a wide-ranging species with a complex life history and to use stable isotope measurements in vertebrae as a means of resolving complexity introduced into the ¹⁴C chronology by ontogenetic shifts in diet and habitat. Stable isotopes provided useful trophic position information; however, validation of age estimates was confounded by what may have been some combination of the dietary source of carbon to the vertebrae, large-scale movement patterns, and steep ¹⁴C gradients with depth in the eastern North Pacific Ocean.     

Insights into vertebral band pair deposition rate in the juvenile common thresher shark (Alopias vulpinus) in the northeastern Pacific Ocean

Validation of band pair deposition rates in elasmobranch vertebrae is essential for accurate age estimation using band pair counting techniques. We present a validation study of the vertebral band pair deposition rate for juvenile common thresher sharks Alopias vulpinus in the northeastern Pacific Ocean (NEPO) using tag and recapture with oxytetracycline (OTC) injection. A total of 14 juvenile A. vulpinus marked with OTC from 1998 through 2013 were recaptured with times at liberty ranging from 1.08 to 3.81 years with an average of 2.14 years (±0.97 years standard deviation, SD). Shark size ranged from 80 to 128 cm fork length (LF) at the time of OTC injection and from 112 to 168 cm LF for those measured at recapture. The slopes of the relationships between band pairs post OTC and years at liberty for each reader ranged from 0.84 to 0.95, slightly lower than the 1.0 slope expected from annual band pair formation. These findings preliminarily support previous age and growth assumptions based on a one band pair per year deposition rate. However, high variation in band pair deposition rates between samples, coupled with regression slopes falling just under one band pair per year, indicates that further investigation is needed to refine band pair deposition rate estimates.     

Effects of insulin-like growth factor-I, corticosterone, and 3,3', 5-tri-iodo-L-thyronine on glycosaminoglycan synthesis in vertebral cartilage of the clearnose skate, Raja eglanteria.

Effects of insulin-like growth factor-I (IGF-I), corticosterone, and triiodothyronine (T(3)) on in vitro growth of vertebral cartilage of the clearnose skate, Raja eglanteria, were investigated. Uptake of [(35)S]sulfate in cultured vertebrae was used to characterize glycosaminoglycan (GAG) synthesis and cartilage growth. IGF-I significantly enhanced cartilage growth when concentrations of 1.28 and 12.8 nM were present in the culture system. Corticosterone significantly inhibited vertebral GAG synthesis at concentrations of 1, 10, and 100 nM. This effect was markedly pronounced in cartilage exposed to 1 and 10 nM corticosterone, in which GAG synthesis was virtually ceased. In contrast, T(3) (0.75, 7.5, and 75.0 nM) had no significant effect on sulfate uptake. These data suggest that IGF-I and corticosteroids may play important roles in regulating skeletal growth of elasmobranchs, as they appear to do in other vertebrates. While T(3) does not appear to exert an immediate, direct effect on vertebral growth, it may still influence elasmobranch chondrogenesis over longer culture periods or indirectly through other regulatory pathways. Thus, further information is necessary to characterize the role of thyroid hormones in the skeletal growth of these fishes. The present study is the first in vitro investigation on the hormonal regulation of elasmobranch cartilage growth. As such, the methods described herein provide a useful technique for examining these physiological processes. J. Exp. Zool. 284:549-556, 1999.     

A modified decalcification technique for enhancing growth bands in deep-coned vertebrae of elasmobranchs

A technique for enhancing growth bands on vertebrae was assayed using 477 centra from blackmouth catsharks, Galeus melastomus. Specimens were caught at depths between 200 and 800 meters off the southern Portuguese coast (Algarve) during an IPIMAR research cruise in August 1994. Total lengths ranged from 12 to 71 cm. Standard vertebral ageing techniques, such as silver nitrate impregnation, haematoxylin staining, grinding and shading failed to produce clearly contrasting bands. A method solely based on vertebra decalcification with nitric acid (5%) yielded the most satisfactory results. This technique provided clear band differentiation and was used by two independent readers. The analysis of differences in band counts between readers was based on: percent of disagreements, percent of disagreement by length group, index of average percent error and Wilcoxon's signed ranks test. The majority of differences corresponded to only one band of disagreement and disagreements have been found to increase with length, which stresses the need for a larger sampling effort on the larger and older individuals. The index of average percent error was estimated as 4.6% and no statistical differences were detected between readers' results.     

Bomb dating and age validation using the spines of spiny dogfish (Squalus acanthias)

Bomb radiocarbon has previously been used to validate the age of large pelagic sharks based on incorporation into vertebrae. However, not all sharks produce interpretable vertebral growth bands. Here we report the first application of bomb radiocarbon as an age validation method based on date-specific incorporation into spine enamel. Our results indicate that the dorsal spines of spiny dogfish, Squalus acanthias, recorded and preserved a bomb radiocarbon pulse in growth bands formed during the 1960s with a timing which was very similar to that of marine carbonates. Using radiocarbon assays of spine growth bands known to have formed in the 1960s and 1970s as a dated marker, we confirm the validity of spine enamel growth band counts as accurate annual age indicators to an age of at least 45 year. Radiocarbon incorporation into northeast Atlantic dogfish spines occurred in similar years as those in the northwest Atlantic and northeast Pacific, although the amount of radiocarbon differed in keeping with the radiocarbon content of the different water masses. Published reports suggesting that Pacific dogfish are longer lived and slower growing than their Atlantic counterparts appear to be correct, and are not due to errors in interpreting the spine growth bands. Radiocarbon assays of fin spine enamel appears to be well suited to the age validation of sharks with fin spines which inhabit the upper 200 m of the ocean.     

Age and Growth of Endangered Smalltooth Sawfish (Pristis pectinata) Verified with LA-ICP-MS Analysis of Vertebrae

Endangered smalltooth sawfish (Pristis pectinata) were opportunistically sampled in south Florida and aged by counting opaque bands in sectioned vertebrae (n = 15). Small sample size precluded traditional age verification, but fish collected in spring and summer had translucent vertebrae margins, while fish collected in winter had opaque margins. Trends in Sr:Ca measured across vertebrae with laser ablation-inductively coupled plasma-mass spectrometry corresponded well to annual salinity trends observed in sawfish estuarine nursery habitats in south Florida, thus serve as a chemical marker verifying annual formation of opaque bands. Based on that finding and assumptions about mean birth date and timing of opaque band formation, estimated age ranged from 0.4 y for a 0.60 m total length (TL) male to 14.0 y for a 4.35 m TL female. Von Bertalanffy growth parameters computed from size at age data were 4.48 m for L_∞, 0.219 y⁻¹for k, and −0.81 y for t₀. Results of this study have important implications for sawfish conservation as well as for inferring habitat residency of euryhaline elasmobranchs via chemical analysis of vertebrae.     

A quantitative study of morphological and histological changes in the skeleton of Salmo salar during its anadromous migration

A quantitative study of morphological and histological changes in the skeleton (cranial bones and vertebrae) of adult Atlantic salmon Salmo salar during its anadromous migration was performed in order to specify various aspects of its skeletal biology in relationship to this migration. At the beginning of the ascent, there was no morphological difference in the cranial bones between males and females. As the spawning season approached, males showed marked secondary sexual characters particularly allometric breeding growth of some bones of the skull. The histological analysis of the vertebral bone tissue along the vertebral axis showed that bone compacity and number of trabeculae vary depending on their localization on the vertebral axis. Moreover, bone compacity decreased significantly with the sexual maturation of the fish whereas the number of trabeculae grew in both sexes. Thus, the vertebrae (like scales) represent an important source of calcium and other elements during anadromous migration in Atlantic salmon.     

Age and growth of the round stingray Urobatis halleri at Seal Beach, California

The age and growth of the round stingray Urobatis halleri was determined using vertebral sections from animals collected at Seal Beach, California from 2002 to 2005. Annual periodicity was validated from U. halleri injected with oxytetracycline and maintained in captivity over a 2 year period ( n = 7). The coefficients estimated by the von Bertalanffy growth model were the disc width asymptote ( W _D∞) (286 mm for males and 224 mm for females) and K (0·09 year⁻¹ for males and 0·15 year⁻¹ for females). The age structure of the population consisted of mostly older, mature males and females. Age at maturity was estimated at 3·80 years for females and 3·75 years for males, and the maximum assessed age was 14 years old. Males were more numerous than females throughout the year; however, from May to September, females outnumbered males. The U. halleri age and growth coefficients were comparable to other species in the family Urolophidae. Based on the seasonality and age structure of this population, Seal Beach offers warm-water refuge for U. halleri of reproductive maturity, and the U. halleri at Seal Beach may garner some behavioural thermoregulation benefit.     

Building an evolutionary innovation: Differential growth in the modified vertebral elements of the zebrafish Weberian apparatus

The Weberian apparatus, a complex assemblage of greatly modified vertebral elements, significantly enhances hearing within Otophysi. Ultimately we are interested in investigating the genetic mechanisms responsible for the origin, development and morphological diversification of these vertebral elements in the Weberian apparatus of otophysan fishes. However, a necessary first step involves identifying changes in growth of this region as compared with the vertebrae from which these modified elements purportedly derive. Using an ontogenetic series of the zebrafish, Danio rerio, we collected growth data for specific elements within the Weberian apparatus, including neural arches, ribs, and parapophyses. These data are compared to both serially homologous structures in posterior thoracic vertebrae (which act as internal controls) and vertebral elements from the same axial levels in three other non-otophysan teleosts. Significant differences in growth rate were found among serially homologous structures, as well as at equivalent axial levels in different species. Uniform changes in growth rates (in which all structures derived from a specific somite were equally affected) were not found, suggesting precise targeting of morphological change to specific structures. The variation in growth of anterior vertebrae in and among species was greater than expected. This variation in growth rates created developmental patterns unique to each species. Such patterns of growth may help illuminate the specific heterochronic mechanisms required for the origin and subsequent morphological diversification of the Weberian apparatus. This morphological diversity is exemplified by the multitude of forms seen in the cypriniform Weberian apparatus. Understanding patterns of growth in discrete elements of the Weberian apparatus allows us to hypothesize as to the specific developmental changes, likely constituting differences in gene expression in pathways involved in bone and cartilage differentiation, responsible for this morphological diversity.     

Analysis of variability in vertebral morphology and growth ring counts in two Carcharhinid sharks

Inter- and intra-regional variations in vertebrae morphology and growth increment counts (band counts) were analyzed for two carcharhinid shark species, Carcharhinus plumbeus (n = 10) and C. limbatus (n = 11). Five sequential vertebrae were removed from the cervical region, above the branchial chamber and posterior to the chondrocrainium, and thoracic region, below the first dorsal fin. Dorsal–ventral height, medial–lateral breadth, and caudal–cranial length were measured for each sampled vertebra. Results indicate no significant difference in vertebral morphology within a sampled region of the vertebral column. However, a significant difference in vertebral morphology was noted between regions for both shark species, with thoracic vertebrae consistently larger than cervical vertebrae. A sub-set of three vertebrae was taken from each sampled region of each shark for sectioning and counting of growth increments. Analyses of growth increment counts by two readers indicated no significant difference in band counts within and between sampled regions.     

Seasonal and environmental codes in the chemistry of the scales of the hoki Macruronus novaezelandiae

Hoki (Macruronus novaezelandiae: Merluccidae) scales do not have conventional annual (annuli) marks, but when observed by transmitted light showed light and dark bands of the type produced by intra-annual variation in the growth rate of the internal lamellae of the scale. The calcium, fluorine, strontium and zinc content of the hoki scale was measured with a proton microprobe and showed cycles in elemental composition that could be interpreted as seasonal.     

Differential segmental growth of the vertebral column of the rat (Rattus norvegicus)

Despite the pervasive occurrence of segmental morphologies in the animal kingdom, the study of segmental growth is almost entirely lacking, but may have significant implications for understanding the development of these organisms. We investigate the segmental and regional growth of the entire vertebral column of the rat (Rattus norvegicus) by fitting a Gompertz curve to length and age data for each vertebra and each vertebral region. Regional lengths are calculated by summing constituent vertebral lengths and intervertebral space lengths for cervical, thoracic, lumbar, sacral, and caudal regions. Gompertz curves allow for the estimation of parameters representing neonatal and adult vertebral and regional lengths, as well as initial growth rate and the rate of exponential growth decay. Findings demonstrate differences between neonatal and adult rats in terms of relative vertebral lengths, and differential growth rates between sequential vertebrae and vertebral regions. Specifically, relative differences in the length of vertebrae indicate increasing differences caudad. Vertebral length in neonates increases from the atlas to the middle of the thoracic series and decreases in length caudad, while adult vertebral lengths tend to increase caudad. There is also a general trend of increasing vertebral and regional initial growth and rate of growth decay caudad. Anteroposterior patterns of growth are sexually dimorphic, with males having longer vertebrae than females at any given age. Differences are more pronounced (a) increasingly caudad along the body axis, and (b) in adulthood than in neonates. Elucidated patterns of growth are influenced by a combination of developmental, functional, and genetic factors.