Daily and sub-daily otolith increments of larval and juvenile walleye pollock, Theragra chalcogramma (Pallas), as validated by alizarin complexone experiments

Walleye pollock (Theragra chalcogramma) were reared from eggs to the juvenile life stage to study daily increment formation in the sagittae otoliths, which are routinely used for age and growth analyses. The apparent deposition of sub-daily growth increments becomes problematic for determining fish age from the late larval stage throughout the juvenile (young-of-the-year) development stage. Otolith marking experiments were conducted to determine interpretation criteria to differentiate between daily and sub-daily increments. Immersion of larval and transforming walleye pollock in 25 mg/l of alizarin complexone (ALC) for 6 h once a week produced a fluorescent mark on the day of staining. Evidence of six well defined and equally spaced increments counted between the weekly ALC marks validated the deposition of daily increments. The daily increments gradually increased in width as the fish/otolith grew. The criteria for determining the presence of sub-daily increments between the daily increments were (1) weak optical definition and (2) a sudden change in incremental distance that lasted for one or two increments and were approximately <0.5 μm in width. Growth problems that occurred during the experiments were identified on otoliths as reductions in daily incremental widths and optical definition, which continued for several days. Otoliths from field-collected fish have also shown similar changes in daily increment properties during the juvenile stage, which may be an indicator of an environmental influence. The criteria for defining different increment types help to resolve our current age determination issues for late larval and early juvenile stage walleye pollock from the Gulf of Alaska.     

Otolith development and daily increment formation in laboratory-reared larval and juvenile black-spot tuskfish <Emphasis Type="Italic">Choerodon schoenleinii</Emphasis>

Microstructures of lapilli were examined for reared larvae and juveniles of black-spot tuskfish Choerodon schoenleinii. Lapilli of larvae at 1 day after hatching have one diffuse and obscure ring with an otolith radius of 4.3 ± 0.50 μm (mean ± SD, N = 8). The slope and intercept of the regression between the number of days after hatching and increment counts did not differ significantly from one and zero, respectively, indicating that lapillus increments were formed on a daily basis after hatching. There was an ontogenetic shift in the relative values of somatic and otolith growth, which corresponded to the transition from pelagic larvae to settlement stage. Simultaneously, the daily increment width reached the maximum value. These findings suggest that age at maximum value of increment width can be used as an indicator of the planktonic larval duration while settlement mark is not found. Since ontogenetic shift in the relationship between otolith radius and body size was observed, back-calculation of somatic growth in black-spot tuskfish using the otolith radius during the early life stages should be analyzed with caution, and the method requires further validation.     

Otolith microstructure of brown sole <Emphasis Type="Italic">Pseudopleuronectes herzensteini</Emphasis>: validation of daily ring formation and the occurrence of microstructure denoting metamorphosis

Brown sole Pseudopleuronectes herzensteini larvae and juveniles were reared to validate daily otolith ring formation. At 15°C, a check (a distinct ring) formed on the sagittae and lapilli at 6 days after hatching, and clear increments regularly formed outside the check. For both otoliths, the relationship between the number of days after hatching and number of increments was linear, and the slope of the line was approximately 1; therefore, daily formation was validated. At 12°C, the check formed on the lapillus 8 days after hatching. Accessory primordia (AP) began forming on the sagittae of metamorphosing larvae, and the shape of the sagittae became complicated. AP were not formed on the lapillus; concentric rings were formed throughout larval and juvenile stages. Wide and obscure increments formed on the lapilli during metamorphosis (metamorphosing zone, MZ). Based on MZ, concentric rings that have formed on the lapilli of juveniles can be separated into larval and juvenile rings. The morphs of large juveniles’ lapilli were bilaterally asymmetric, and the blind-side lapilli were most suitable for otolith microstructure analysis. This study provides fundamental information for otolith microstructure analysis in wild brown sole.     

Otolith microstructure of chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis>: formation of sea entry check and daily deposition of otolith increments in seawater conditions

To apply otolith microstructure to examination of age and growth of juvenile chum salmon Oncorhynchus keta inhabiting coastal waters, formation of otolith increments was investigated for juveniles reared in a seawater aquarium and in net pens. In all otoliths examined, a distinctive check was formed at the time of sea entry of the fish. The deposition of otolith increments after the check was daily for rearing both in the aquarium (57 days) and in the net pens (26 days). Check formation associated with sea entry was also observed in otoliths of juvenile salmon collected 1 km off the coast of Shari, Hokkaido, Japan. Transmitted light observation of otoliths of those fish revealed a transition in otolith increment appearance from dark to light. Otolith Sr: Ca ratio remarkably changed from a low to a high level, coinciding with the transition in otolith appearance. It is suggested that the transition was associated with individual sea entry. This study demonstrated that the check and/or transition associated with sea entry are applicable to a benchmark for otolith increment counts of juvenile chum salmon inhabiting coastal waters.     

Validation of otolith increment daily periodicity in captive juvenile sablefish (Anoplopoma fimbria) experimentally immersed in strontium chloride (SrCl2)

We used a chemical marking experiment to validate the daily periodicity of otolith increment deposition in juvenile sablefish, Anoplopoma fimbria. The sagittal otoliths of 26 live juvenile sablefish were marked twice by immersion in saltwater baths containing elevated levels of strontium chloride (SrCl₂) during June–August of their first year of life (age 0). The number of otolith increments detected between strontium bands from three readings (median 15–20) was compared to the number of days between marking events (15–17 days). Median discrepancies between otolith increment counts and days between strontium bands were small (mean 1.1, S.E. 0.81, n = 20) and suggest that otolith increment counts from age 0 juvenile sablefish may provide a useful proxy for daily age. However, median discrepancies in June (mean 4.7, S.E. 0.65, n = 9 otoliths) were significantly (^*P ≤ 0.05) higher than those in July (mean −1.8, S.E. 0.40, n = 6) and August (mean −1.8, S.E. 0.66, n = 5). Increment banding patterns were more difficult to identify in otoliths marked in June (mean sablefish size 84.9 mm FL) because of changes in the structure of the sagittae associated with the formation of accessory primordia.     

Validation of otolith daily increments for larval and juvenile Japanese halfbeak Hyporhamphus sajori

Daily ring formation was verified by examining the growth of the marginal increment on sagittal otoliths of larval and juvenile Japanese halfbeak Hyporhamphus sajori. The relationship between age (x) and number of increments (y) is y = 1.0x + 2.0. The first daily increment was formed during the 2 days before hatching. The relationship between the number of rings deposited after the alizarin complexone (ALC) mark (x) and the number of increments (y) is given by: y = 1.0x − 0.2. The index of completion of the marginal increment was 99 ± 4.1% (mean ± S.D.) at 04:00, and from 24 ± 4.4% at 08:00 it increased with time of day and reached 98 ± 3.6% until the next day at 04:00. Growth of the incremental zone started a few hours after sunrise prior to which the discontinuous zone seemed to be formed. Light rhythms tend to be one of the most important factors for the formation of the marginal increment on otoliths. Based on the relationships between time of day and the marginal increment on otoliths, it would be possible to estimate the predation time for specimens retrieved from stomach contents, and also clearer analysis of the growth history immediately prior to the sampling time.     

Age determination and growth of juveniles of the European hake,Merluccius merluccius (L., 1758), in the northern Tyrrhenian Sea (NW Mediterranean)

The aim of the present study was to provide an estimation of growth of juvenile European hake, Merluccius merluccius (L., 1758) (OSTEICHTHYES; MERLUCCIIDAE), by means of the analysis of otolith daily increments. Hake specimens were collected during trawl surveys carried out in the northern Tyrrhenian Sea (NW Mediterranean). The sagittae were removed from hakes ≤20 cm total length. Left otoliths were ground and polished to obtain thin frontal sections. Otolith microstructure was analysed under a compound green light-polarising microscope. A power curve with intercept was fitted to the length-age data to describe the growth of M. merluccius. According to the growth curve, a mean length of 18 cm was reached at the end of the first year of life. The validation of the otolith increment periodicity was performed by means of two indirect methods.     

Age validation,growth and annual reproductive cycle of chub mackerel <Emphasis Type="Italic">Scomber japonicus</Emphasis> off the waters of northern Kyushu and in the East China Sea

The age and growth of chub mackerel Scomber japonicus collected from the East China Sea and the northern waters off Kyushu between June 2000 and June 2001 were determined by observing the otolith surface after dipping it in xylene. The translucent and opaque zones on the otolith surface were identified, and the number of translucent zones was counted. Monthly changes in the frequency of fish with translucent zones on the otolith margin, and in the marginal increments, indicated that the translucent zones were formed between April and June. The seasonal pattern of annulus formation on the otolith became clear by observing the otoliths of fish with known ages, and the otolith formation in wild fish was consistent with that of fish with known ages. The mean gonadosomatic index of male and female fish was high from March to May, and spawning females were observed from mid-March to mid-May. The estimated ages were 1–5 years for males and 1–6 years for females. The von Bertalanffy growth parameters did not significantly differ between male and female. The model was obtained as FL _t=406×{1−exp[−0.372×(t+1.68)]     

Elemental analysis of otoliths of Japanese anchovy: trial to discriminate between Seto Inland Sea and Pacific stock

The ability to discriminate local stocks of Japanese anchovy Engraulis japonicus was assessed based on data from four elements (K, Na, P, and Sr) using an electron probe micro analyzer (EPMA) and data from three elements (Ba, Mn, and Sr) using inductively coupled plasma mass spectrometry (ICP-MS) from the otoliths of 40 anchovy (23.6–47.0 mm body length). Anchovy were caught at three sites (Aki-nada, Hiuchi-nada, and Osaka Bay) in the Seto Inland Sea, and one site (Kuroshio extension) in the Pacific Ocean in 2002. In order to discriminate different spawning grounds, EPMA data from the core portion (from core to 30 μm in the core-posterior axis) were used. Results showed that it was difficult to discriminate between the Seto Inland Sea and the Pacific anchovy by EPMA data. Conversely, it was possible to discriminate between the Seto Inland Sea and the Pacific anchovy by ICP-MS data from bulk otoliths. Our results showed that Mn contents of otoliths using ICP-MS discriminate between spawning grounds most, and Ba and Sr discriminate less. The difference in elemental compositions in anchovy otoliths between the Seto Inland Sea and the Pacific Ocean might be reflected by cumulative experienced elemental composition of ambient sea water during life history between the Seto Inland Sea and the Pacific anchovy.     

Age, growth and life history of gunnel, Pholis fangi, in the Yellow Sea

We examined the formation of annuli by marginal observations on otoliths of gunnel (Pholis fangi) in the Yellow Sea to validate the age determination method and to derive the growth equation covering from larval to adult stages. Gunnels, ranging from 46 to 173 mm in total length, were collected by a bag net fishery from the western coastal waters off Korea from November 1998 to October 1999. Marginal observations indicated that the translucent zone (annual mark) on adult otolith was formed during the winter, whereas the opaque zone was formed during the summer. However, a translucent zone was formed between May and June in juvenile otoliths. This false ring was formed when the fish transited from the inshore pelagic life of larvae to the offshore bottom life of juveniles. The observed maximum age was 58 months. Using observed length-at-monthly age, growth in length was expressed by von Bertalanffy growth curve; L_t = 144.0 (1 − e^{−0.11 (t+0.43)}). P. fangi spawned in winter recruit to inshore, and grow quickly in the nursery habitats in spring. Gunnel inhabit the bottom offshore area during the summer season, and reappear inshore thereafter.     

Responses in growth rate of larval northern anchovy (Engraulis mordax) to anomalous upwelling in the northern California Current

We examined variability in growth rate during the larval stage of northern anchovy (Engraulis mordax) in response to physical and biological environmental factors in 2005 and 2006. The onset of spring upwelling was anomalously delayed by 2–3 months until mid‐July in 2005; in contrast, spring upwelling in 2006 began as a normal year in the northern California Current. Larval and early juvenile E. mordax were collected in August, September, and October off the coast of Oregon and Washington. Hatch dates ranged from May to September, with peaks in June and August in 2005 and a peak in July in 2006, based on the number of otolith daily increments. Back‐calculated body length‐at‐age in the June 2005 hatch cohort was significantly smaller than in the August 2005 cohort, which had comparable growth to the July 2006 cohort. Standardized otolith daily increment widths as a proxy for seasonal variability in somatic growth rates in 2005 were negative until late July and then changed to positive with intensification of upwelling. The standardized increment width was a positive function of biomass of chlorophyll a concentration, and neritic cold‐water and oceanic subarctic copepod species sampled biweekly off Newport, Oregon. Our results suggest that delayed upwelling in 2005 resulted in low food availability and, consequently, reduced E. mordax larval growth rate in early summer, but once upwelling began in July, high food availability enhanced larval growth rate to that typical of a normal upwelling year (e.g., 2006) in the northern California Current.     

茜素络合物浸泡标记秦岭细鳞鲑发眼卵及仔鱼耳石

采用茜素络合物(ALC)在秦岭细鳞鲑(Brachymystax lenok tsinlingensis)发眼卵和仔鱼阶段进行浸泡标记试验,为确定合适浸泡浓度和持续浸泡时间,试验设置了6个的浓度组和4个时间梯度组.结果显示:在荧光显微镜下观察被浸泡的仔鱼和经浸泡处理的发眼卵而出膜后的仔鱼,其耳石都出现橘红色荧光标记.浸泡...     

Prevalence of stocked whitefish in River Kemijoki,Finland, inferred by micro X‐ray fluorescence analysis of otoliths

Micro X‐ray fluorescence (µ‐XRF) analysis of otoliths was evaluated as a method to estimate the proportion of stocked one‐summer‐old whitefish Coregonus lavaretus L. in catches of adult fish (n = 20) ascending the River Kemijoki to spawn. Laser ablation inductively coupled plasma mass spectrometry (LA‐ICP‐MS) analysis was applied as control. Polished otoliths were scanned with µ‐XRF to obtain strontium maps that were used to infer visually the provenance of the whitefish. Thirteen of the fish showed signs of being stocked as one‐summer‐old fingerlings. LA‐ICP‐MS was applied to determine the elemental composition in a spot outside the core of the otolith. The results were largely consistent with the visual inspection of the µ‐XRF mapped otoliths. In conclusion, µ‐XRF mapping successfully identified whitefish stocked as one‐summer‐old fingerlings. The vast majority of whitefish returning to the River Kemijoki to spawn were stocked fish.     

Somatic growth and otolith microstructure of larval and juvenile pacific cod <Emphasis Type="Italic">Gadus macrocephalus</Emphasis>

ABSTRACT:   Microstructures of sagittae and lapilli were examined in relation to somatic growth for reared larvae and juveniles of Pacific cod. The Laird–Gompertz model was fitted to the daily age and somatic growth relationship. Growth increments were deposited on a daily basis in both kinds of otoliths, with a check formed at hatching. Two subsequent checks and an accessory primordia (AP) occurred in the sagittae. The lapillus was adequate for increment width measurement through the early life stages. Sagittal and frontal plane of sagitta was adequate for measurement in the pre-AP and post-AP formation stages, respectively. The shift of desirable plane was caused by changes in otolith and increment shapes with AP formation. Back-calculated total lengths using the biological intercept method did not significantly differ with certain body lengths ( P  > 0.05), suggesting validity of back-calculation in this species. Using the back-calculated total length, morphological and ecologic changes that seemed to affect checks and AP formations are discussed.     

Early life history of the black anglerfish Lophius budegassa Spinola, 1807 in the Mediterranean Sea using otolith microstructure

The early life history of the black anglerfish, Lophius budegassa was investigated by otolith (lapilli) increment analysis. Samples of demersal juvenile L. budegassa ranging from 54 to 196 mm total length were collected during bottom trawl surveys in the central Adriatic Sea. By counting increments presumed to be deposited daily in the lapillar otoliths, 88 specimens of L. budegassa were successfully aged. Age estimates of juveniles ranged between 79 and 204 days, indicating that probably the pelagic phase of this species is relatively short and settlement occurs at less than 3 months of life. The analysis of check marks in the core area of lapilli enabled us to determine the period of endogenous feeding, which would last between 15 and 24 days after hatching. Back-calculated hatching dates and, consequently, the spawning season of L. budegassa in the Adriatic Sea was spread over a long period, lasting at least from February to June. The length at age relationship gave an estimate of mean growth rate of approximately 0.8–1.02 mm/day, indicating a faster growth rate of 0+ juveniles L. budegassa than previously thought. The implications of these findings on age estimates discrepancies between previous ageing studies on L. budegassa carried out using different calcified structure (sagittae or illicia) are discussed.     

Can calcein and alizarin complexone be used for double immersion marking of juvenile qingbo <Emphasis Type="Italic">Spinibarbus sinensis</Emphasis>?

Calcein (CAL) from 50 to 250 mg/l and alizarin complexone (ALC) from 100 to 300 mg/l were used for double immersion marking of juvenile qingbo Spinibarbus sinensis. With the exception of the scales, double immersion for 24 h produced detectable double marks in otoliths (sagittas and asterisci), barbs, fin rays (dorsal, pectoral, ventral, anal, and caudal), and fin spines (dorsal, pectoral, ventral, and anal) after 90 days in a laboratory growth experiment. Green fluorescent rings produced by CAL were considerably closer to the inside of the bony structures (including otoliths, barbs, fin rays, and fin spines) than red fluorescent rings produced by ALC. Sagittas, asterisci, and barbs showed acceptable fluorescent marks at higher concentrations (250 mg/l CAL and ≥ 200 mg/l ALC, ≥ 200 mg/l CAL and 300 mg/l ALC, 150–250 mg/l CAL and 250–300 mg/l ALC, respectively). Fin rays and fin spines treated by 200 mg/l CAL and 250 mg/l ALC and 250 mg/l CAL and 300 mg/l ALC simultaneously had both acceptable CAL and ALC marks. There was no statistically significant difference on the survival or growth of marked fish compared to the controls throughout the experiment (p > 0.05). The results suggest that double immersion with CAL and ALC is suitable for double mass-marking of juvenile S. sinensis, and these double marks are useful in the experimental development of biological research or restocking methodologies.     

Seasonal changes in otolith and somatic growth in age-0 Pacific saury <Emphasis Type="Italic">Cololabis saira</Emphasis>

We evaluated the seasonal changes in otolith and somatic growth of age-0 Pacific saury Cololabis saira in 223 fish collected between June and November 2002. We calculated the age in days of each individual by measuring otolith growth increments under a scanning electron microscope. The age was correlated with body length and otolith radius. We also observed seasonal changes in the rate of increase in body length and otolith radius and in the pattern of otolith growth. Until August, both body length and otolith radius increased with age. Thereafter, the otolith radius continued to increase, whereas the rate of somatic growth decreased. As a result, the ratio of otolith radius to body length increased. After August, the percentage of otoliths with unreadable increments on their edge increased due to the formation of hyaline zones. Otoliths grew both radially and in thickness until July, but gradually stopped growing in thickness after August. Beginning in October, more than 80% of otoliths only grew radially. After August, the otolith not only continued growing but the morphological growth pattern also changed.     

Growth of Northwest Iberian juvenile hake estimated by combining sagittal and transversal otolith microstructure analyses

Daily growth of Atlantic juvenile hake from Northwest Iberia has been estimated employing a new approach combining analyses of transversal and sagittal sections of the otoliths along the ventral radius. Age of juvenile hake ranging from 3 to 25 cm collected during a spring 2002 survey was estimated. Somatic growth followed a power fit: Fish size (TL) = 3.3254*age^0.7336 (r² = 0.87, p < 0.001, n = 76), yielding an average individual growth rate of 0.66 mm/day (±0.06). The growth model indicates that after a year's life a juvenile can reach 25 cm. Otolith ventral radius ranged from 401 to 1842 μm and daily increments were between 104 and 387. Fish growth and otolith growth were closely related (r² = 0.92 p < 0.001, n = 76). These first results of daily growth rates for the Southern stock corroborate the fast-growth hypothesis of this species. The evolution of increment widths from hatch dates onwards reveals important seasonal growth peaks during July–August and October–November. A comparison with prior data and discussion is also presented in the light of recent work on hake juveniles and tagging-recapture experiences.     

Validation of the otolith increment deposition ratio using alizarin marks in juveniles of the sparid fishes, Diplodus vulgaris and D. puntazzo

Deposition rate of otolith increments was validated by two immersions in alizarin complexone 10 days apart in juvenile Diplodus vulgaris and D. puntazzo. Alizarin complexone produced a well-defined scarlet band on the otoliths of all individuals. One increment:one day ratio was found for both species.     

Growth of juvenile chub mackerel Scomber japonicus in the western North Pacific Ocean: with application and validation of otolith daily increment formation

The growth of juvenile chub mackerel Scomber japonicus collected in the western North Pacific Ocean in 2007 and 2009 was examined based on the evidence of otolith daily increment formation in captive specimens. There was a significant difference in the relationship between known age and number of increments in the frontal and sagittal planes. Repeated markings on the otolith using Alizarin complexone and the coefficient of variation in number of increments suggest that the increments in the frontal plane of the otolith are more suitable for age estimation than those in the long and short axes of the sagittal plane. The increments in the frontal plane formed daily, and the first ring was usually deposited 3 days after hatch. Age of wild juveniles ranged from 24 to 211 days after hatch based on the frontal plane method. The estimated hatching periods of specimens ranged from February to June, but the April-hatched specimens were collected throughout the sampling periods of 2007 and 2009. The Gompertz growth model showed a difference in growth pattern in specimens between 2007 and 2009. The juveniles in 2009 appeared to grow more quickly than those in 2007 until summer, but thereafter the 2009 specimens seemed to grow more slowly.