Growth and age validation in high-Antarctic fish

The growth of three demersal species dominant in biomass and abundance in the Weddell Sea, Chionodraco myersi (channichthyid), Trematomus eulepidotus (nototheniid) and Dolloidraco longedorsalis (artedidraconid), was studied on specimens caught in summer. Ages were determined using ground and polished otolith sections and an image analysis system to measure the otolith radius and distances of each growth increment to the otolith core. Ageing precision and accuracy have been estimated. Indirect validation methods, based on the increment growth pattern, were applied to support the annual periodicity of the increments. Due to the lack of some age classes, back-calculated age-length keys were determined and used to fit the von Bertalanffy growth parameters by sex and species. The determined values showed sexual dimorphism, with females having faster growth rates, except in D. longedorsalis. The growth performance index was between 1 and 2, as in other high-Antarctic fish. For T. eulepidotus, age and growth data were available, but this is the first report for C. myersi and D. longedorsalis. Moreover, this is the first time that accuracy and precision have been estimated for high-Antarctic fish age determination. Accepted: 20 March 2000     

Age determination in common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis>: history,relative utility of ageing structures,precision and accuracy

The common carp Cyprinus carpio is one of the most widely-distributed freshwater fishes in the world. Due to its value for conservation and fisheries in several native/translocated areas of distribution and its detrimental effects on the aquatic ecosystem in most invasive areas, robust age-based population dynamics models are required for successful management of this species. The present study provides a global review of age determination in carp, including a historical account of ageing methods, an assessment of the relative utility of ageing structures, and an evaluation of precision and accuracy (i.e. validation) of age estimates. Historically, scales were by far the most widely-employed structure, followed by the operculum, otolith, dorsal spine, vertebra and fin ray. However, in countries where carp is categorised as ‘high risk’ of impact, use of alternative structures to the scale was predominant. Causal criteria analysis showed scales and opercula to provide inconsistent evidence for successful annulus identification/counting, whereas consistent evidence was found for otoliths, dorsal spines, vertebrae and (pectoral) fin rays. Precision was always above reference thresholds for scales, whereas for otoliths, dorsal spines and fin rays was in several cases below. Accuracy was addressed sporadically and mostly in high-risk countries. It is suggested that dorsal spines or pectoral fin rays should be used in lieu of scales as non-lethal ageing structures, and otoliths (or vertebrae, pending more research) otherwise, and that validation should always be attempted as part of the set-up of more appropriate ageing protocols and use of correct terminology.     

High resolution bomb dating for testing the accuracy of age interpretations for a short-lived pelagic fish, the Atlantic herring

Although stock assessments of Atlantic herring (Clupea harengus harengus) in the northwest Atlantic have relied on tens of thousands of annual age determinations each year for more than 20 years, recent analyses have suggested that there may have been systemic ageing error. Tracking of dominant year-classes and otolith exchanges confirmed the presence of substantial ageing bias among some readers, although these approaches could not be used to identify an accurate set of ages (if any). We applied bomb radiocarbon in a high resolution dating approach targeted at the 1962 year-class to assess ageing accuracy by multiple age readers and laboratories. Although bomb radiocarbon age validation studies are typically restricted to long-lived species, the availability of archived otoliths through the 1960s and 1970s made herring an ideal candidate for an approach targeted at a single year-class, and allowed the extent of any ageing error to be quantified. The results clearly demonstrated that current age reading practices under-aged fish >6 yr of age by up to 45%, although younger fish were aged accurately. Age underestimation was due to incorrect annulus interpretation rather than non-annual otolith growth. By focusing on the period of most rapid radiocarbon increase (1962), the margin of uncertainty around the targeted bomb radiocarbon ages was reduced to 0.66 yr. This study represents the first time the bomb dating method has reached sub-annual accuracy, which makes it well suited to the age validation of short lived fish species. The use of the targeted approach has considerable promise for improving the accuracy of other bomb radiocarbon studies without the problematic assumptions associated with curve estimation and environmental effects.     

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.     

New insights into estimating the age of old Scots pine from increment cores with stem rot

Trunk inner rot is a common phenomenon in some old-growth pine dominated forests, making it impossible to determine tree age by counting annual rings. We compared the efficiency of five methods to estimate the age of hollow pine trees (Pinus sylvestris L.). Our main aims were to select the best-performing method and to test whether the age of the tree or the proportion of rot influences the accuracy of estimation. We used full increment cores (reaching the pith or within 1 cm of it) from 100 trees (54–562 years old) collected in northern Sweden and simulated rotten centres of three different sizes in order to test the methods. The lowest error rates were obtained when less than a third of the sample was missing (down to 5.0 % error rate), and by using a method based on the growth pattern of a set of healthy trees. Using linear extrapolation of the mean radial growth led to large overestimates (up to three times the number of absent rings) with error rates up to 27.3 %. We also found that the performance of all methods was reduced in cores from older trees. Our main conclusion is that non-linear methods should be preferred for age estimation of hollow pines. We also argue that more precision in the age estimation could be gained already in the field by collecting multiple cores from rotten trees or by developing alternative coring methods.     

A comparison of two methods for the microscopic determination of age at death.

The precision and accuracy of the Kerley and Ahlqvist-Damsten microscopic methods of age determination are compared. Both methods were applied to the same sample of 40 femoral thin sections of documented age at death. The results indicate that (1) both methods can be used with equal precision, as suggested by comparable observer errors; and (2) the Kerley method produces overall more accurate age estimates. The low previously published standard error of the Ahlqvist-Damsten method (6.71 years) apparently results from the uneven age distribution and small size (20) of their sample.     

Otolith annulus validation and variables influencing false annuli formation in dorsal spines of saugeye

Marginal increment analysis is a common technique for validating formation of a single annual growth ring on an ageing structure. False annuli can form on ageing structures when environmental variables affect growth of a fish, potentially resulting in age estimation bias. Therefore, validating ageing structures is essential to ensure that accurate and precise age estimates are collected when assessing fish population dynamics. Saugeye (Sander vitreus, [Mitchill, 1818]) and S. Canadensis, [Griffith and Smith, 1834]) are highly valued sport fish that are stocked across the Midwest United States. Using marginal increment analysis, we confirmed that a single annulus was formed yearly in otoliths of juvenile saugeye, however two annuli formed in dorsal spines in a single year. Timing of the first annulus formation in both otoliths and dorsal spines was completed after a slow growth period during winter (otoliths forming in April; dorsal spines forming in March). The second annulus (false annulus) that formed during August in dorsal spines did not form in otoliths. To understand what environmental factors may influence the false annulus to form, we collected monthly water temperatures and percent empty stomachs of juvenile saugeye. The highest water temperatures of the year occur during July and August, which resulted in saugeye seeking thermal refuge and affecting feeding habits. Mean monthly temperature and percent empty stomachs were positively correlated, so during times of high temperatures foraging rates declined, suggesting the formation of false annulus on dorsal spines of juvenile saugeye. This study demonstrates how thermal stress affected accuracy of non‐lethal aging structures and further emphasizes the need for age validation studies prior to using non‐lethal ageing structures to estimate age for a particular species from different aquatic systems.     

Biological clocks as age estimation markers in animals: a systematic review and meta-analysis

Various biological attributes associated with individual fitness in animals change predictably over the lifespan of an organism. Therefore, the study of animal ecology and the work of conservationists frequently relies upon the ability to assign animals to functionally relevant age classes to model population fitness. Several approaches have been applied to determining individual age and, while these methods have proved useful, they are not without limitations and often lack standardisation or are only applicable to specific species. For these reasons, scientists have explored the potential use of biological clocks towards creating a universal age-determination method. Two biological clocks, tooth layer annulation and otolith layering have found universal appeal. Both methods are highly invasive and most appropriate for post-mortem age-at-death estimation. More recently, attributes of cellular ageing previously explored in humans have been adapted to studying ageing in animals for the use of less-invasive molecular methods for determining age. Here, we review two such methods, assessment of methylation and telomere length, describing (i) what they are, (ii) how they change with age, and providing (iii) a summary and meta-analysis of studies that have explored their utility in animal age determination. We found that both attributes have been studied across multiple vertebrate classes, however, telomere studies were used before methylation studies and telomere length has been modelled in nearly twice as many studies. Telomere length studies included in the review often related changes to stress responses and illustrated that telomere length is sensitive to environmental and social stressors and, in the absence of repair mechanisms such as telomerase or alternative lengthening modes, lacks the ability to recover. Methylation studies, however, while also detecting sensitivity to stressors and toxins, illustrated the ability to recover from such stresses after a period of accelerated ageing, likely due to constitutive expression or reactivation of repair enzymes such as DNA methyl transferases. We also found that both studied attributes have parentally heritable features, but the mode of inheritance differs among taxa and may relate to heterogamy. Our meta-analysis included more than 40 species in common for methylation and telomere length, although both analyses included at least 60 age-estimation models. We found that methylation outperforms telomere length in terms of predictive power evidenced from effect sizes (more than double that observed for telomeres) and smaller prediction intervals. Both methods produced age correlation models using similar sample sizes and were able to classify individuals into young, middle, or old age classes with high accuracy. Our review and meta-analysis illustrate that both methods are well suited to studying age in animals and do not suffer significantly from variation due to differences in the lifespan of the species, genome size, karyotype, or tissue type but rather that quantitative method, patterns of inheritance, and environmental factors should be the main considerations. Thus, provided that complex factors affecting the measured trait can be accounted for, both methylation and telomere length are promising targets to develop as biomarkers for age determination in animals.     

Age determination of corvina reina (Cynoscion albus) in the Gulf of Nicoya, Costa Rica, based on examination and analysis of hyaline zones, morphology and microstructure of otoliths

This is the first age determination study for Cynoscion albus , a large tropical sciaenid, using otolith morphology and daily increment analysis. The practicality of both methods for age determination is illustrated by their consistent estimates of age and von Bertalanffy growth parameters. The daily increment analysis was used to validate the surface readings. An alternative, otolith morphometrics, is shown to hold promise for rapid prediction of fish age. Two multivariate linear regression models using gross otolith dimensions can estimate the age of C. albus to within 1 year. Growth parameter estimates are: from surface readings: L =127.5cm with K =0.12; from daily increment readings: L = 122.1 cm with K =0.17. Implications for the stock assessment of tropical fish using size instead of age are discussed.     

Separating vascular and neuronal effects of age on fMRI BOLD signals

Accurate identification of brain function is necessary to understand the neurobiology of cognitive ageing, and thereby promote well-being across the lifespan. A common tool used to investigate neurocognitive ageing is functional magnetic resonance imaging (fMRI). However, although fMRI data are often interpreted in terms of neuronal activity, the blood oxygenation level-dependent (BOLD) signal measured by fMRI includes contributions of both vascular and neuronal factors, which change differentially with age. While some studies investigate vascular ageing factors, the results of these studies are not well known within the field of neurocognitive ageing and therefore vascular confounds in neurocognitive fMRI studies are common. Despite over 10 000 BOLD-fMRI papers on ageing, fewer than 20 have applied techniques to correct for vascular effects. However, neurovascular ageing is not only a confound in fMRI, but an important feature in its own right, to be assessed alongside measures of neuronal ageing. We review current approaches to dissociate neuronal and vascular components of BOLD-fMRI of regional activity and functional connectivity. We highlight emerging evidence that vascular mechanisms in the brain do not simply control blood flow to support the metabolic needs of neurons, but form complex neurovascular interactions that influence neuronal function in health and disease.This article is part of the theme issue ‘Key relationships between non-invasive functional neuroimaging and the underlying neuronal activity’.     

Validation of determination of lead (Pb) in blood by electrothermal atomic absorption spectrometry (ETAAS) on the basis of interlaboratory comparison data

ISO 15189 standard establishes a requirement to periodically revalidate analytical methods for the determination of trace elements like Pb in blood, as conditions change and technical advances are made. The aim of this study was to revalidate an electrothermal atomic absorption spectrometry (ETAAS) method for determination of Pb in blood over the microrange 25–35 μg/dL, on the basis of historical results of interlaboratory comparison programmes. Precision and inaccuracy were estimated by analysis of records of an external quality control programme for Pb (PICC-PbS). The precision and inaccuracy values obtained were both less than 5%, highly satisfactory in view of the validation requirement that precision and inaccuracy be less than 10%. These findings demonstrate the effectiveness of this new validation methodology, which does not require any disruption of the laboratory's routine activity, and which can be used even if the method in question has not been validated previously at that laboratory.     

Development of an alternative ageing technique for blue shark (Prionace glauca) using the vertebra

Accurate determination of age is essential for the comprehensive assessment and management of exploited shark populations. Enhancement of growth bands is necessary to accurately and efficiently determine age. However, most traditional techniques do not describe an efficiency of a series of procedures and the detailed protocol for different‐sized sharks. We describe a simple and highly successful technique for ageing vertebrae of blue shark (Prionace glauca) that we refer to as “burn method”, derived from the “break and burn method” and “shadowing method”—a simple procedure requiring an alkaline treatment to clean the vertebral centra and a burning treatment to enhance growth band visualization. We described optimal times for sharks of precaudal length of 50, 100, 150, and 200 cm as 44.9, 88.7, 134.0, 183.5 s of alkaline treatment, and 6.8, 8.9, 10.2, and 11.5 min of burning treatment; both longer and shorter exposure times resulted in higher failed sample frequencies. Using our burn method, it is possible to do ageing a sample from a wide angle, with the reader able to obtain information on growth bands from different perspectives. To critique different technique efficiencies, the index of average percent error (IAPE) and mean coefficient of variation (CV) are compared for independent readers using the burn method and other techniques (silver nitrate impregnation and the unstained shadowing methods). The precision of the burn method (IAPE, 4.1%; CV, 5.7%) was similar to that of silver nitrate impregnation (IAPE, 5.8%; CV, 8.2%) and unstained shadowing method (IAPE, 8.3%; CV, 11.8%). For younger specimens, the IAPE and CV of the burn method were lower than those of the other techniques, but, compared with other ageing methods, precision decreased for older sharks. We demonstrate that the burn method to be accurate for age determination of blue sharks, especially for specimens with less than 10 bands, but recommend simultaneous use of other methods, such as thin sectioning (the most common for structure‐based ageing) and bomb carbon dating, to accurately determine the age of older individuals.     

Quantification of the age-pigment lipofuscin in known-age octopus (Octopus pallidus): A potential tool for age determination

Stylet increment analysis (SIA) is the key method to age octopus, however, currently it is not reliable for all species. The suitability of the age-pigment lipofuscin as an alternative ageing method for octopus was examined. To determine the relationship between age and lipofuscin known-age octopus (Octopus pallidus) were reared in the laboratory from hatching to eight months old. Twenty-eight individuals at three different ages (3, 6 and 8 months old) were collected for lipofuscin analysis. The first two age groups (n = 5 each) were reared under ambient temperatures, while the oldest group (n = 18) was reared under three different controlled temperature regimes (n = 6 per treatment). For comparison, five wild O. pallidus were also collected for lipofuscin analysis and aged using SIA. Lipofuscin was analysed in the brain tissue and quantified at a commercial ageing centre using standard histological methods. Lipofuscin granules were clearly discernable in the brain tissue, and there was a strong exponential relationship between age and lipofuscin (R² = 0.86). Lipofuscin concentration was not related to sex, temperature or body weight in same-age individuals. Except for one individual, the predicted age of the wild animals, based on the relationship between lipofuscin and age, was close to the age determined using SIA. This study is the first to report lipofuscin in an octopus species and shows that lipofuscin has excellent potential as an alternative ageing method for octopus. This research will have important applications for species which cannot be reliably aged using current ageing methods.     

Ageing with fisheries length-frequency data, using information about growth

A new approach to break down fisheries length-frequency data into age groups, using information on relative growth rate with age, is introduced. The method was verified with data used to test previously published methods. Good starting values for the mean lengths of the fish with age in the statistical analysis of distribution mixtures are often critical. Incorporation of relative growth rate information significantly improves the precision of the ageing procedure even when starting values are less accurate.     

Epigenetic age prediction

Advanced age is the main common risk factor for cancer, cardiovascular disease and neurodegeneration. Yet, more is known about the molecular basis of any of these groups of diseases than the changes that accompany ageing itself. Progress in molecular ageing research was slow because the tools predicting whether someone aged slowly or fast (biological age) were unreliable. To understand ageing as a risk factor for disease and to develop interventions, the molecular ageing field needed a quantitative measure; a clock for biological age. Over the past decade, a number of age predictors utilising DNA methylation have been developed, referred to as epigenetic clocks. While they appear to estimate biological age, it remains unclear whether the methylation changes used to train the clocks are a reflection of other underlying cellular or molecular processes, or whether methylation itself is involved in the ageing process. The precise aspects of ageing that the epigenetic clocks capture remain hidden and seem to vary between predictors. Nonetheless, the use of epigenetic clocks has opened the door towards studying biological ageing quantitatively, and new clocks and applications, such as forensics, appear frequently. In this review, we will discuss the range of epigenetic clocks available, their strengths and weaknesses, and their applicability to various scientific queries.     

Determination of Zygosity in Adult Chinese Twins Using the 450K Methylation Array versus Questionnaire Data

Previous studies have shown that both single nucleotide polymorphisms (SNPs) and questionnaires-based method can be used for twin zygosity determination, but few validation studies have been conducted using Chinese populations. In the current study, we recruited 192 same sex Chinese adult twin pairs to evaluate the validity of using genetic markers-based method and questionnaire-based method in zygosity determination. We considered the relatedness analysis based on more than 0.6 million SNPs genotyping as the golden standards for zygosity determination. After quality control, qualified twins were left for relatedness analysis based on identical by descent calculation. Then those same sex twin pairs were included in the zygosity questionnaire validation analysis. Logistic regression model was applied to assess the discriminant ability of age, sex and the three questions in zygosity determination. Leave one out cross-validation was used as a measurement of internal validation. The results of zygosity determination based on 65 SNPs in 450k methylation array were all consistent with genotyping. Age, gender, questions of appearance confused by strangers and previously perceived zygosity consisted of the most predictable model with a consistency rate of 0.8698, cross validation predictive error of 0.1347. For twin studies with genotyping and\or 450k methylation array, there would be no need to conduct other zygosity testing for the sake of costs consideration.     

基于树木起源、立地分级和龄组的单木生物量模型

以马尾松(Pinus massoniana)和落叶松(Larix)的大样本实测资料为建模样本,以独立抽取的样本为验证样本,把样本按起源、立地和龄组进行分级,采用与材积相容的两种相对生长方程,分普通最小二乘和两种加权最小二乘,对地上部分总生物量、地上各部分生物量和地下生物量进行模型拟合和验证,使用决定系数、均方根误差、总相对误差和估计精度等8项统计量对结果进行分析。结果表明:两个树种地上部分总生物量,立地分类方法,模型的拟合结果和适用性都最优;马尾松VAR模型较优,而落叶松CAR模型较好;两种加权最小二乘方法,在建模样本和验证样本中表现得不一致。在建模样本中,加权回归2(权重函数1/f^0.5)略优于加权回归1(权重函数1/y^0.5),但在验证样本中,加权回归1却明显优于加权回归2。而同时满足建模样本拟合结果最优和验证样本检验结果最优的组合中,只有加权回归1。两个树种地上部分各分量生物量,模型拟合结果和适用性,均为干材最优,树叶最差、树枝和树皮居中,样本分类、模型类型和加权最小二乘方法对干材生物量的影响,规律和地上部分总生物量相同;样本分类、模型类型和加权最小二乘方法的最优组合,用验证样本检验的结果,总相对误差树枝不超过±10.0%,树皮不超过±5.0%,树叶马尾松不超过±30.0%,落叶松不超过±20.0%。两个树种地下部分(根)生物量,样本按龄组分类方法,模型拟合结果最优,与材积相容的模型总体上优于与地上部分总生物量相容模型。     

Age and growth studies of chondrichthyan fishes: the need for consistency in terminology, verification, validation, and growth function fitting

Validated age and growth estimates are important for constructing age-structured population dynamic models of chondrichthyan fishes, especially those which are exploited. We review age and growth studies of chondrichthyan fishes, using 28 recent studies to identify areas where improvements can be made in describing the characteristics of ageing structures (both traditional and novel) utilized to estimate ages of sharks, rays, and chimaeras. The topics identified that need consistency include the: (1) terminology used to describe growth features; (2) methods used to both verify and validate age estimates from chondrichthyan calcified structures, especially edge and marginal increment analyses; and (3) the functions used to produce and describe growth parameters, stressing the incorporation of size at birth (L ₀) and multiple functions to characterize growth characteristics, age at maturity and longevity.     

Do reef corals age?

Hydra is emerging as a model organism for studies of ageing in early metazoan animals, but reef corals offer an equally ancient evolutionary perspective as well as several advantages, not least being the hard exoskeleton which provides a rich fossil record as well as a record of growth and means of ageing of individual coral polyps. Reef corals are also widely regarded as potentially immortal at the level of the asexual lineage and are assumed not to undergo an intrinsic ageing process. However, putative molecular indicators of ageing have recently been detected in reef corals. While many of the large massive coral species attain considerable ages (>600 years) there are other much shorter‐lived species where older members of some populations show catastrophic mortality, compared to juveniles, under environmental stress. Other studies suggestive of ageing include those demonstrating decreased reproduction, increased susceptibility to oxidative stress and disease, reduced regeneration potential and declining growth rate in mature colonies. This review aims to promote interest and research in reef coral ageing, both as a useful model for the early evolution of ageing and as a factor in studies of ecological impacts on reef systems in light of the enhanced effects of environmental stress on ageing in other organisms.