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.     

Age and growth information available from the otoliths of the Hawaiian snapper,Pristipomoides filamentosus

The otoliths of tropical fish may provide important life history information incorporated within their structural and chemical constituents. All three otoliths (sagitta, lapillus, asteriscus) of the tropical fish Pristipomoides filamentosus were examined internally by Scanning Electron Microscope methods to observe micro-increments and externally to determine three dimensional structure. It was discovered that the sagitta contained four cores and that the plane chosen to be sectioned for micro-increment enumeration could result in errors if more than one core were transversed. The medial cross sectional plane was consequently resolved to effer the most accurate micro-increment counts. Obserations of lapilli also revealed micro-increments and subsequent counts were closely correlated to those detected in the sagittae. The visualization of increments made it feasible to assess age and evolve a growth model. In addition, sagitta weight was found to be related to growth rate and may provide a quick estimate of relative growth. Chemical analyses of otoliths for stable isotopes and Sr/Ca ratios all suggested that an individual fish inhabited warmer waters as it became older. A combination of otolith structural and chemical information can provide age and growth data which is essential to the calculation of accurate population parameters.     

A comparison of different structures and methods for estimating age of northern-form Dolly Varden <Emphasis Type="Italic">Salvelinus malma malma</Emphasis> from the Canadian Arctic

Assessment of anadromous northern-form Dolly Varden Salvelinus malma malma in the Western Canadian Arctic requires reliable methods for estimating ages. Additionally, conservation efforts warrant determining whether fin rays provide a non-lethal alternative to otoliths. Precision and bias of whole and sectioned otoliths, and sectioned pectoral and pelvic fin rays were examined. Two age readers with different levels of experience ageing this species read each structure three times. Coefficient of variation (CV) was calculated to measure precision, and age bias plots were created for each method of preparation/structure within and between readers. The experienced reader demonstrated the highest precision with sectioned otoliths (CV = 1.6 %) followed by whole otoliths (CV = 4.2 %) while pectoral and pelvic fins were the lowest, CV = 7.7 % and 7.5 %, respectively. The age bias plot showed little difference between whole and sectioned otoliths, although greater imprecision/bias was evident for whole otoliths at age ≥9. Compared to otoliths, fin rays produced younger age estimates starting at 5 years; however, pelvic fins were more biased towards younger estimates than pectoral fins. The less experienced reader had greater inconsistencies, tending to overage younger and underage older samples for all methods compared to the more experienced reader, underscoring the importance of experience when estimating age for this species. We conclude that both types of fin rays are a poor non-lethal alternative to otoliths for fish ≥5 years and recommend an experienced ager could use whole otoliths up to age 8 and sectioned otoliths for fish ≥9 years (>500 mm fork length).     

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.     

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.     

Aging three-spined sticklebacks Gasterosteus aculeatus: comparison of estimates from three structures

In order to assess the accuracy and reliability of age estimates from calcified structures in the three-spined stickleback Gasterosteus aculeatus, we evaluated intra and inter-reader repeatability from three structures: otoliths, gill covers and pelvic spines). Average age estimates were also compared between the structures. The overall intra-reader repeatability of age estimates were highest for otoliths (69%), lowest for gill covers (53%) and intermediate for spine cross-sections (63%). Although four of the seven readers had the highest intra-reader repeatability score for spine cross-sections, the inter-reader variance in this structure was much higher than in others. Otoliths were the easiest in terms of their pre-analysis treatment and exchange of materials (as digital images) between readers. In addition, otoliths are more well-studied compared with the other structures with respect to their development through ontogenesis; hence, age estimates based on otoliths should be the most reliable. Therefore, our recommendation is that whenever possible, analysis of otoliths should be the preferred approach for aging G. aculeatus.     

Precision of age determination from otoliths,opercular bones,scales and vertebrae in the threatened freshwater snakehead,Channa punctata (Bloch, 1793)

Ages were estimated from otoliths, opercular bones, scales, and vertebrae of 514 specimens of Channa punctata from three Indian rivers – the Ganga, Gomti, and Yamuna – to evaluate the potential bias of age estimates between readers and between pairs of aging structures. Standard procedures were followed to prepare and study the age structures. Among all structures, otoliths showed highest (97.4%) values of agreement between readers' age estimates followed by the opercular bones (89.5%), scales (84.2%) and vertebrae (78.9%). Because of the highest percentage of agreement and lowest average percentage of error (0.05%) and coefficient of variation (0.76%) values between readers, otoliths were considered the most suitable aging structure for C. punctata. When otoliths ages were compared with those of the other structures viz., opercular bones, scales and vertebrae, percentage of agreement was found highest between estimates (90.8%) of otoliths and opercular bones.     

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.     

Growth and otolith microstructure of cod (Gadus morhua L.) larvae

Cod larvae from Irish Sea stocks were reared under differentgrowth conditions, and the otolith growth and increment formationexamined in sagittae and lapilli. Otolith increments were firstdeposited around the time of hatching and increment counts,on average, reflected larval age. The growth rate of fish larvaereared in different sized tanks was significantly different(P < 0.001), but there was no detectable effect on incrementformation. Otolith size was independent of larval size for individuals<5 mm in length. In larvae >6 mm, larger, faster growingindividuals had larger and faster growing otoliths.     

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.     

Residence time and drift patterns of larval June sucker Chasmistes liorus in the lower Provo River as determined by otolith microstructure

Estimates of age derived from daily ring counts from otoliths and capture rates of larval June sucker Chasmistes liorus were used to determine the relationship between discharge rates of the Provo River and residence time and patterns of larval drift. During 1997, larval drift occurred over a 22 day period when discharge rates were low (mean ±s.d. 3·2 ± 0·0 m³ s^?1). In 1998, larval drift occurred in two separate events over a 40 day period. Discharge was higher during the first larval drift period (19 days; 24·8 ± 1·3 m³ s^?1) and lower during the second larval drift period (17 days; 7·0 ± 0·9 m³ s^?1). In 1997, no larval fish were collected at the lowermost transect on the Provo River (nearest Utah Lake), and few larvae >21 days of age were found. During the first drift period of 1998, larval C. liorus were collected at all transects, and mean age of larvae collected between upstream and downstream transects increased by c. 7 days. During the second drift period of 1998, only a few were collected in the lowermost transects, and age did not increase with proximity to the lake. Patterns in catch and age distribution of larval C. liorus in the lower Provo River suggest that recruitment failure occurs during the larval drift period in years with insufficient discharge to transport larvae into the lake.     

Effects of growth rates on the otolith increments deposition rate in capelin larvae (Mallotus villosus)

Otolith microstructure analysis was applied to known age capelin larvae (Mallotus villosus) in order to examine the formation of daily increments. In two validation experiments, newly hatched yolk sac larvae were stocked into eight 10 m³ plastic bags where environmental conditions were kept as natural as possible. The bags were terminated after 35-79 days, the surviving larvae were collected and the otoliths were analysed. Survival in the bags varied between 39 - 71% with average individual length growth rates of 0.25 mm day^- 1. The ages of most larvae were underestimated and the accuracy in age estimation was generally low. Highest accuracy was found for fast growing larvae. On average, the larvae started to form increments about 12 days after hatching, and the increment width increased with age and/or length of the larvae. Larvae showing low body growth rates had fast otolith growth relative to body length.     

The back-calculation of size and growth from otoliths: Validation and comparison of models at an individual level

Multiple internal and external tagging experiments tested the applicability of five back-calculation models (Biological Intercept, Modified-Fry, Body Proportional Hypothesis, Time-Varying Growth, and an Age-Effects model) as predictors of individual growth trajectories of two marine cleaning gobies, Elacatinus evelynae and E. prochilos, that were raised in aquaria under conditions that resulted in variable growth. Mixed-effect model analyses of longitudinal records of otoliths and somatic growth collected at the individual level revealed that E. evelynae and E. prochilos produced daily increments on their otoliths for up to two months post-settlement and that the Modified-Fry model provided the most accurate size-at-age estimates despite the presence of age, growth and time-varying growth effects in the dataset. Very large errors in predicted size were produced by the Age-Effects model. The four other back-calculation models all provided slightly biased estimates of back-calculated size-at-age, with the Modified-Fry model providing the least biased estimates. Regardless of bias, both experimental and theoretical evidence indicated that back-calculated size was a better proxy of fish length than otoliths radius. Relationships between fish length and otoliths radius were allometric at the level of individuals, which explained why the Modified-Fry model performed better. However, this allometry was undetectable at the population level. This study represents the first attempt to validate modern back-calculation models using longitudinal data collected and analysed at the individual level. Our results suggest the use of the Modified-Fry model as a conservative approach in routine back-calculations of fish size at age from otoliths.     

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 effect of sire selection on the response of lambs to vaccination with irradiated Trichostrongylus colubriformis larvae

Dineen, J. K. and Windon, R. G. (1980). The effect of sire selection on the response of lambs to vaccination with irradiated Trichostrongylus colubriformis larvae. International Journal for Parasitology10: 189–196. Rams selected for responsiveness and unresponsiveness to vaccination with irradiated T. colubriformis larvae at an early age were mated to unselected random bred ewes. Progeny were vaccinated with 20,000 irradiated larvae at 8 and 12 weeks of age, given anthelmintic treatment at 16 weeks and challenged with 20,000 normai larvae at 17 weeks. The results, based on wether worm counts and ewe faecal egg counts, showed significant differences between responder and non-responder progeny. There was a significant correlation between worm counts and faecal egg counts of half-sibs from the same sire group. The occurrence of globule leucocytes was inversely related to worm burdens of wether progeny, however, no clear relationship was found with eosinophils. In vitro lymphocyte stimulation using T. colubriformis L₃ antigen, concanavalin A and lipopolysaccharide showed that statistically defined responder progeny, pooled from both responder and non-responder sire groups, gave higher responses than non-responder lambs after vaccination. The results confirm that genetically-determined factors are involved in the response of lambs to vaccination at an early age, and indicate that rapid genetic progress may be achieved in the type of mating usually carried out under fleld conditions.     

The effect of selection of both sire and dam on the response of F1 generation lambs to vaccination with irradiated Trichostrongylus colubriformis larvae

Windon R. G. and Dineen J. K. (1981). The effect of selection of both sire and dam on the response of F₁ generation lambs to vaccination with irradiated Trichostrongylus colubriformis larvae. International Journal for Parasitology11: 11–18. Rams and ewes, tested for responsiveness to vaccination with irradiated T. colubriformis larvae at an early age, were mated on the basis of responder × responder and non-responder × non-responder. Progeny were vaccinated at 8 and 12 weeks of age with 20,000 irradiated larvae, treated with anthelmintic at 16 weeks and challenged with 20,000 normal larvae at 17 weeks. Faecal egg counts of progeny from responder matings were significantly lower than progeny from non-responders, and within each mating type, ewe lambs had markedly lower egg counts than ram lambs. The level of circulating complement-fixing antibodies to T. colubriformis larval extract were inversely related to egg counts. Thus, ewe progeny from responder matings had the highest serum antibody levels, non-responder ram progeny had the lowest levels and responder rams and non-responder ewes had similar intermediate levels. In vitro responses of cells stimulated with T. colubriformis L₃ antigen were greater in progeny from responder matings, whereas responses to bacterial lipopolysaccharide were higher in progeny from non-responder matings. The results confirm that the response to vaccination at an early age is genetically determined, and show that the response of progeny is most vigorously expressed when both sires and dams have been selected.     

Validation of gum-line recession as a reliable technique to age tigers

Like other members of the Panthera genus, tigers are threatened throughout their range. Given their conservation status research focused on their population dynamics in the wild is needed, including population growth, lifespan, and breeding success, which requires reliable estimates of age. Current techniques to estimate age in tigers are either not reliable and/or difficult to apply in the field. Gum-line recession accurately estimates age for mountain lions, but has never been tested for tigers. The goal of this study was to determine whether gum-line recession of the upper canine teeth is a reliable indicator of age in tigers and if so, to define the equation that enables age estimation. We measured gum-line recession in 12 individuals of known ages and performed linear regression analysis to investigate the validity of this technique for tigers. We found a strong relationship between gum-line recession and age, where the model provided reliable age estimates for animals within 1-year age classes in 10 out of 12 tigers measured, providing increased accuracy over current methods.     

Pelagic larval duration and population connectivity in New Zealand triplefin fishes (Tripterygiidae)

The relationship between pelagic larval duration (PLD) and population connectivity in marine fishes has been controversial, but most studies to date have focused on tropical taxa. Here, we examine PLD in 11 species of triplefin fishes from a temperate environment in the Hauraki Gulf, New Zealand, to describe daily increment patterns and settlement marks in the otoliths. The formation of daily increments was validated using larvae of known age and tetracycline marking of settled juveniles. Settlement mark identity was verified by comparing total increment counts from otoliths of recently settled fishes with PLD counts from post-settlement fishes. A similar pattern of three groups of increments across the otolith was found in all specimens examined. The settlement mark was similar in all species and occurred as a sharp drop in increment width within the area of transition in optical density. PLD was lengthy, compared to species of triplefins from elsewhere, and ranged between 54.4 ± 1.7 SE days in Bellapiscis lesleyae to 86.4 ± 2.6 SE days in Forsterygion malcolmi. Variation in PLD within species was high but did not mask interspecific differences. PLD was not phylogenetically constrained, as sister species differed significantly in PLD. PLD was compared with genetic population connectivity for eight of the study species using mitochondrial gene flow data from Hickey, Lavery, Hannan, Baker, Clements. Mol Ecol 18:680–696 (2009). The observed lack of correlation between PLD and gene flow suggests that dispersal is limited by other factors, such as larval behaviour and the availability of settlement habitat.     

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.     

Age and growth of the Antarctic dragonfish Parachaenichthys charcoti (Pisces, Bathydraconidae) from the southern Scotia Arc

The Antarctic dragonfish Parachaenichthys charcoti is commonly found in shelf waters of islands in the southern Scotia Arc. Its northern congener P. georgianus is distributed on the shelves of South Georgia and the South Sandwich Islands. Biological information on P. charcoti is limited and restricted largely to reproductive traits and feeding habits. The present study fills this gap, providing the first data on age and growth of this species by otolith reading. Age was estimated in juveniles and adults by counting annuli on otoliths. Age of early juveniles was determined by microincrement counts (considered to be daily rings). Age estimates of juveniles and adults ranged from 1 to 9?years, with a high percentage agreement between readings and low values of counting variability indices APE (3.3?%) and CV (4.7?%). Age of early juveniles ranged from 160 to 204?days, all showing an evident check at 32–35?days of age, tentatively linked to the first larval exogenous feeding. The estimated values of von Bertalanffy growth parameters L _∞ and k were 53.55?cm and 0.22, respectively, and the index of growth performance P was 2.33. Hatching in P. charcoti takes place in late winter, from August to September. Compared to other notothenioids inhabiting the Seasonal Pack-ice Zone, P. charcoti is a relatively fast growing species with moderate longevity, sharing several biological characteristics with its sister species P. georgianus.