Age determination manual of the Alaska Fisheries Science Center Age and Growth Program

The Age and Growth Program at the Alaska Fisheries Science Center is tasked with providing age data in order to improve the basic understanding of the ecology and fisheries dynamics of Alaskan fish species. The primary focus of the Age and Growth Program is to estimate ages from otoliths and other calcified structures for age-structured modeling of commercially exploited stocks; however, the program has recently expanded its interests to include numerous studies on topics ranging from age estimate validation to the growth and life history of non-target species. Because so many applications rely upon age data and particularly upon assurances as to their accuracy and precision, the Age and Growth Program has developed this practical guide to document the age determination of key groundfish species from Alaskan waters. The main objective of this manual is to describe techniques specific to the age determination of commercially and ecologically important species studied by the Age and Growth Program. The manual also provides general background information on otolith morphology, dissection, and preparation, as well as descriptions of methods used to measure precision and accuracy of age estimates. This manual is intended not only as a reference for age readers at the AFSC and other laboratories, but also to give insight into the quality of age estimates to scientists who routinely use such data.

Age, growth and mortality of the toadfish, Halobatrachus didactylus (Schneider, 1801) (Pisces: Batrachoididae), in the Bay of Cádiz (southwestern Spain)

Age, growth and mortality of the toadfish, Halobatrachus didactylus, were determined by examination of the whole sagittal otoliths of fish sampled in the Bay of Cádiz (southwestern Spain) from March 1999 to March 2000. A total of 844 specimens (425 males, 416 females, and 3 of indeterminate sex), ranging from 95 to 470 mm in total length were examined. Eighty-nine percent of the otoliths could be read allowing an age estimation. The opaque zone was formed between April and May coincident with the maximum reproductive peak, while the translucent zone formed mainly in summer-fall (June to December). Maximum ages for males and females were 12 and 10 years, respectively. The samples were dominated by 2- to 6-year-old specimens. Males matured at an age of approximately 2 years and females at 3 years. Fish total length and otolith radius were closely related. The von Bertalanffy growth curve was used to describe growth. The parameters were derived from back-calculated length-at-age. Significant differences in the growth parameters were found between sexes. Although the growth analysis revealed that this species is slow-growing, males reached larger sizes than females. Females appeared to experience higher natural mortality rates than males.

Use of skeletochronological analysis to estimate the age of leatherback sea turtles Dermochelys coriacea in the western North Atlantic

Although growth rate and age data are essential for leatherback management, estimates of these demographic parameters remain speculative due to the cryptic life history of this endangered species. Skeletochronological analysis of scleral ossicles obtained from 8 captive, known-age and 33 wild leatherbacks originating from the western North Atlantic was conducted to characterize the ossicles and the growth marks within them. Ages were accurately estimated for the known-age turtles, and their growth mark attributes were used to calibrate growth mark counts for the ossicles from wild specimens. Due to growth mark compaction and resorption, the number of marks visible at ossicle section tips was consistently and significantly greater than the number visible along the lateral edges, demonstrating that growth mark counts should be performed at the tips so that age is not underestimated. A correction factor protocol that incorporated the trajectory of early growth increments was used to estimate the number of missing marks in those ossicles exhibiting resorption, which was then added to the number of observed marks to obtain an age estimate for each turtle. A generalized smoothing spline model, von Bertalanffy growth curve, and size-at-age function were used to obtain estimates of age at maturity for leatherbacks in the western North Atlantic. Results of these analyses suggest that median age at maturation for leatherbacks in this part of the world may range from 24.5 to 29 yr. These age estimates are much greater than those proposed in previous studies and have significant implications for population management and recovery.

Sources of age determination errors for sablefish (Anoplopoma fimbria)

This study was undertaken to resolve problems in age determination of sablefish (Anoplopoma fimbria). Aging of this species has been hampered by poor agreement (averaging less than 45%) among age readers and by differences in assigned ages of as much as 15 years. Otoliths from fish that had been injected with oxytetracycline (OTC) and that had been at liberty for known durations were used to determine why age determinations were so difficult and to help determine the correct aging procedure. All fish were sampled from Oregon southwards, which represents the southern part of their range. The otoliths were examined with the aid of image processing. Some fish showed little or no growth on the otolith after eight months at liberty, whereas otoliths from other fish grew substantially. Some fish lay down two prominent hyaline zones within a single year, one in the summer and one in the winter. We classified the otoliths by morphological type and found that certain types are more likely to lay down multiple hyaline zones and other types are likely to lay down little or no zones. This finding suggests that some improvement could be achieved by detailed knowledge of the growth characteristics of the different types. This study suggests that it may not be possible to obtain reliable ages from sablefish otoliths. At the very least, more studies will be required to under-stand the growth of sablefish otoliths.

Age determination and growth of the night shark (Carcharhinus signatus) off the northeastern Brazilian coast

Age and growth of the night shark (Carcharhinus signatus) from areas off northeastern Brazil were determined from 317 unstained vertebral sections of 182 males (113–215 cm total length [TL]), 132 females (111.5–234.9 cm) and three individuals of unknown sex (169–242 cm). Although marginal increment (MI) analysis suggests that band formation occurs in the third and fourth trimesters in juveniles, it was inconclusive for adults. Thus, it was assumed that one band is formed annually. Births that occur over a protracted period may be the most important source of bias in MI analysis. An estimated average percent error of 2.4% was found in readings for individuals between two and seventeen years. The von Bertalanffy growth function (VBGF) showed no significant differences between sexes, and the model derived from back-calculated mean length at age best represented growth for the species (L∞=270 cm, K=0.11/yr, t0=–2.71 yr) when compared to the observed mean lengths at age and the Fabens’ method. Length-frequency analysis on 1055 specimens (93–260 cm) was used to verify age determination. Back-calculated size at birth was 66.8 cm and maturity was reached at 180–190 cm (age 8) for males and 200–205 cm (age ten) for females. Age composition, estimated from an age-length key, indicated that juveniles predominate in commercial catches, representing 74.3% of the catch. A growth rate of 25.4 cm/yr was estimated from birth to the first band (i.e. juveniles grow 38% of their birth length during the first year), and a growth rate of 8.55 cm/yr was estimated for eight- to ten-year-old adults.

Effects of density-dependence and sea surface temperature on interannual variation in length-at-age of chub mackerel (Scomber japonicus) in the Kuroshio-Oyashio area during 1970–1997

Annual mean fork length (FL) of the Pacific stock of chub mackerel (Scomber japonicus) was examined for the period of 1970–97. Fork length at age 0 (6 months old) was negatively correlated with year-class strength which fluctuated between 0.2 and 14 billion in number for age-0 fish. Total stock biomass was correlated with FL at age but was not a significant factor. Sea surface temperature (SST) between 38–40°N and 141–143°E during April–June was also negatively correlated with FL at age 0. A modified von Bertalanffy growth model that incorporated the effects of population density and SST on growth was well fitted to the observed FL at ages. The relative FL at age 0 for any given year class was maintained throughout the life span. The variability in size at age in the Pacific stock of chub mackerel is largely attributable to growth during the first six months after hatching.

Age and growth of sailfish (Istiophorus platypterus) in waters off eastern Taiwan

Age and growth of sailfish (Istiophorus platypterus) in waters off eastern Taiwan were examined from counts of growth rings on cross sections of the fourth spine of the first dorsal fin. Length and weight data and the dorsal fin spines were collected monthly at the fishing port of Shinkang (southeast of Taiwan) from July 1998 to August 1999. In total, 1166 dorsal fins were collected, of which 1135 (97%) (699 males and 436 females) were aged successfully. Trends in the monthly mean marginal increment ratio indicated that growth rings are formed once a year. Two methods were used to back-calculate the length of presumed ages, and growth was described by using the standard von Bertalanffy growth function and the Richards function. The most reasonable and conservative description of growth assumes that length-at-age follows the Richards function and that the relationship between spine radius and lower jaw fork length (LJFL) follows a power function. Growth differed significantly between the sexes; females grew faster and reached larger sizes than did males. The maximum sizes in our sample were 232 cm LJFL for female and 221 cm LJFL for male.

Maturity, ovarian cycle, fecundity, and age-specific parturition of black rockfish (Sebastes melanops)

From 1995 to 1998, we collected female black rockfish (Sebastes melanops) off Oregon in order to describe their basic reproductive life history and determine age-specific fecundity and temporal patterns in parturition. Female black rockfish had a 50% probability of being mature at 394 mm fork length and 7.5 years-of-age. The proportion of mature fish age 10 or older significantly decreased each year of this study, from 0.511 in 1996 to 0.145 in 1998. Parturition occurred between mid-January and mid-March, and peaked in February. We observed a trend of older females extruding larvae earlier in the spawning season and of younger fish primarily responsible for larval production during the later part of the season. There were differences in absolute fecundity at age between female black rockfish with prefertilization oocytes and female black rockfish with fertilized eggs; fertilized-egg fecundity estimates were considered superior. The likelihood of yolked oocytes reaching the developing embryo stage increased with maternal age. Absolute fecundity estimates (based on fertilized eggs) ranged from 299,302 embryos for a 6-year-old female to 948,152 embryos for a 16-year-old female. Relative fecundity (based on fertilized eggs) increased with age from 374 eggs/g for fish age 6 to 549 eggs/g for fish age 16.

Distribution, age, and growth of young-of-the year greater amberjack (Seriola dumerili) associated with pelagic Sargassum

Patterns of distribution and growth were examined for young-of-the-year (YOY) greater amberjack (Seriola dumerili) associated with pelagic Sargassum in the NW Gulf of Mexico. Seriola dumerili were collected off Galveston, Texas, from May to July over a two-year period (2000 and 2001) in both inshore (<15 nautical miles [nmi]) and offshore zones (15−70 nmi). Relative abundance of YOY S. dumerili (32−210 mm standard length) from purse-seine collections peaked in May and June, and abundance was highest in the offshore zone. Ages of S. dumerili ranged from 39 to 150 days and hatching-date analysis indicated that the majority of spawning events occurred from February to April. Average daily growth rates of YOY S. dumerili for 2000 and 2001 were 1.65 mm/d and 2.00 mm/d, respectively. Intra-annual differences in growth were observed; the late-season (April) cohort experienced the fastest growth in both years. In addition, growth was significantly higher for S. dumerili collected from the offshore zone. Mortality was approximated by using catch-curve analysis, and the predicted instantaneous mortality rate (Z) of YOY S. dumerili was 0.0045 (0.45%/d).

Radiometric validation of age, growth, and longevity for the blackgill rockfish (Sebastes melanostomus)

As nearshore fish populations decline, many commercial fishermen have shifted fishing effort to deeper continental slope habitats to target fishes for which biological information is limited. One such fishery that developed in the northeastern Pacific Ocean in the early 1980s was for the blackgill rockfish (Sebastes melanostomus), a deep-dwelling (300−800 m) species that congregates over rocky pinnacles, mainly from southern California to southern Oregon. Growth zone-derived age estimates from otolith thin sections were compared to ages obtained from the radioactive disequilibria of 210Pb, in relation to its parent, 226Ra, in otolith cores of blackgill rockfish. Age estimates were validated up to 41 years, and a strong pattern of agreement supported a longevity exceeding 90 years. Age and length data fitted to the von Bertalanffy growth function indicated that blackgill rockfish are slow-growing (k= 0.040 females, 0.068 males) and that females grow slower than males, but reach a greater length. Age at 50% maturity, derived from previously published length-at-maturity estimates, was 17 years for males and 21 years for females. The results of this study agree with general life history traits already recognized for many Sebastes species, such as long life, slow growth, and late age at maturation. These traits may undermine the sustainability of blackgill rockfish populations when heavy fishing pressure, such as that which occurred in the 1980s, is applied.

Age validation, growth, mortality, and additional population parameters of the goldband snapper (Pristipomoides multidens) off the Kimberley coast of northwestern Australia

Goldband snapper (Pristipomoides multidens) collected from commercial trap and line fishermen off the Kimberley coast of northwestern Australia were aged by examination of sectioned otoliths (sagittae).A total of 3833 P. multidens, 80–701 mm fork length (98–805 mm total length), were examined from commercial catches from 1995 to 1999. The oldest fish was estimated to be age 30+ years. Validation of age estimates was achieved with marginal increment analysis. The opaque and translucent zones were each formed once per year and are considered valid annual growth increments (the translucent zone was formed once per year between January and May). A strong link between water temperature and translucent zone formation was evident in P. multidens. The von Bertalanffy growth function was used to describe growth from length-at-age data derived from sectioned otoliths.

Age-validation of a leopard shark (Triakis semifasciata) recaptured after 20 years

On 10 July 1999, vertebrae bearing an oxytetracycline (OTC) time mark were retrieved from a tagged leopard shark (Triakis semifasciata) recaptured in San Francisco Bay, CA, after being at liberty for almost 20 years. An additional long-term leopard shark tag return was received in June 2001, for which growth information (but not vertebrae) was obtained. The first recapture is significant in that it represents the longest at-liberty period for an age-validated (OTC-injected) shark, extends and completes age validation for this species, spanning all age classes up to its estimated average maximum age, and provides an example of the persistence of the OTC time mark in an elasmobranch at liberty for almost 20 years. The recaptured leopard shark made in 2001 also provides valuable information on long-term growth from time of release to time of recapture. Findings are documented here so that other researchers are aware that validation is complete for this species, to present pertinent evidence of considerable interannual variability in growth in this species, and to report observations on processing difficulties relating to the ephemeral nature of the 20-yr-old OTC mark.

Demographic assessment of the blue crab (Callinectes sapidus) in Chesapeake Bay using extractable lipofuscins as age markers

The blue crab (Callinectes sapidus) plays an important economic and ecological role in estuaries and coastal habitats from the Gulf of Mexico to the east coast of North America, but demographic assessments are limited by length-based methods. We applied an alternative aging method using biochemical measures of metabolic byproducts (lipofuscins) sequestered in the neural tissue of eyestalks to examine population age structure. From Chesapeake Bay, subsamples of animals collected from the 1998–99 (n=769) and 1999–2000 (n=367) winter dredge surveys were collected and lipofuscin was measured. Modal analysis of the lipofuscin index provided separation into three modes, whereas carapace-width data collected among the same individuals showed two broad modes. Lipofuscin modal analysis indicated that most adults (carapace width >120 mm) were <2 years old. The results indicate that use of extractable lipofuscin can provide a more accurate and better resolved estimation of demographic structure of blue crab populations in the field than size alone.

Age and growth of the estuarine dolphin (Sotalia guianensis) (Cetacea, Delphinidae) on the Paraná Coast, southern Brazil

Teeth of 71 estuarine dolphins (Sotalia guianensis) incidentally caught on the coast of Paraná State, southern Brazil, were used to estimate age. The oldest male and female dolphins were 29 and 30 years, respectively. The mean distance from the neonatal line to the end of the first growth layer group (GLG) was 622.4 ±19.1 μm (n=48). One or two accessory layers were observed between the neonatal line and the end of the first GLG. One of the accessory layers, which was not always present, was located at a mean of 248.9 ±32.6 μm (n=25) from the neonatal line, and its interpretation remains uncertain.The other layer, located at a mean of 419.6 ±44.6 μm (n=54) from the neonatal line, was always present and was first observed between 6.7 and 10.3 months of age. This accessory layer could be a record of weaning in this dolphin. Although no differences in age estimates were observed between teeth sectioned in the anterior-posterior and buccal-lingual planes, we recommend sectioning the teeth in the buccal-lingual plane in order to obtain on-center sections more easily. We also recommend not using teeth from the most anterior part of the mandibles for age estimation. The number of GLGs counted in those teeth was 50% less than the number of GLGs counted in the teeth from the median part of the mandible of the same animal. Although no significant difference (P>0.05) was found between the total lengths of adult male and female estuarine dolphins, we observed that males exhibited a second growth spurt around five years of age. This growth spurt would require that separate growth curves be calculated for the sexes. The asymptotic length (TL∞), k, and t0 obtained by the von Bertalanffy growth model were 177.3 cm, 0.66, and –1.23, respectively, for females and 159.6 cm, 2.02, and –0.38, respectively, for males up to five years, and 186.4 cm, 0.53 and –1.40, respectively, for males older than five years. The total weight (TW)/total length (TL) equations obtained for male and female estuarine dolphins were TW = 3.156 × 10−6 × TL 3.2836 (r=0.96), and TW = 8.974 × 10−5 × TL 2.6182 (r=0.95), respectively.