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.     

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).     

Comparison of scale and otolith age readings for trahira,Hoplias malabaricus (Bloch, 1794), from Paraná River,Argentina

The aim of this work was to compare age determinations and precision using two deposition structures of trahira Hoplias malabaricus (Bloch, 1794): the scales, which are most frequently used, and otoliths (lapilli). The length‐age relationships were obtained with both structures and compared with results from previous studies. The 163 sets of trahira otoliths (lapilli) and scales were 17–46 cm standard length (SL) from Cayastá (Santa Fe) and Islas Lechiguanas (Entre Ríos), Paraná River, Argentina. Three independent readings of each structure were conducted. An age bias plot was performed to compare age estimations from scales and otoliths. To assess the precision of age determinations using both structures, the percent agreement among readers for both structures and the coefficient of variation were calculated (%CV). The age‐length relationships were plotted and fitted with the von Bertalanffy growth function for both structures and compared with previous works. Age readings recorded for scales were lower than those recorded for otoliths for ages above or equal to 3 years. Percent agreement among readers was higher than 80% for otoliths and less than 65% for scales. The%CV obtained for scales was 20% for young fish and 17% for adults. For otoliths the %CV was 7% for young fish and 3% for adults. The %CV obtained for scales was over the recommended limit (>7.6%). The von Bertalanffy parameters for scales were L_inf = 45.80 mm; k = 0.29; t₀ = ?1.34 and for otoliths were L_inf = 40.76 mm; k = 0.39; t₀ = ?1.05. Precision of age estimations assessed from the percent agreement and the coefficient of variation indicates that the scales of the trahira are inappropriate to estimate age in population studies for juvenile and adult specimens.     

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.     

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.     

Validation of scales and otoliths for estimating age of redband trout in high desert streams of Idaho

Studies validating aging structures for rainbow trout are sparse and none have been conducted for redband trout, a common western U.S. sub-species. Oxytetracycline mark-recapture methods (MR), marginal incremental analysis (MIA), and comparisons across multiple populations were used to evaluate the utility of two structures for aging redband trout in high desert streams. We assessed periodicity of annulus formation on scale and otolith samples from all age classes of trout residing in two streams, identified the location of the first annulus on otoliths, and compared age estimates and between-reader coefficient of variation on nine additional streams. The use of MIA successfully validated opaque zone periodicity for fish transitioning from age-0 to age-1, and from age 1 to age 2 in two streams. For fish at liberty 13 and 28 months in the same two streams, MR-derived age estimates from whole and sectioned otoliths were 94–100% accurate for fish from 2–9 years old. Scales were only 77% and 38% accurate for fish at liberty 13 and 28 months, respectively. Between-reader coefficient of variation (CV) for scales was high (11.5%), while CV for sectioned otoliths using whole otoliths as corroboratory structures averaged 2.3%. Scales were thus, an unacceptable aging structure for desert redband trout. Given the confusion in the literature, we suggest that more rigorous research should be conducted to define and explain otolith zone formation.     

Comparing aging precision of calcified structures in shovelnose sturgeon

Age estimates for population analysis must be precise. We assessed the usefulness of pectoral fin rays, sphenoids, opercula, and dorsal scutes of shovelnose sturgeonScaphirhynchus platorynchus (n = 30) as aging structures based on ease of collection, distinctness of annuli, and measures of precision both between and within readers. We also determined how age estimates from paired fin rays of individuals were related (n = 106). Pectoral fin rays generated the highest within‐reader precision (100% within 2 years) followed by sphenoids (58%), opercula (56%), and dorsal scutes (49%). Ages estimated by the pectoral fin ray also had higher between‐reader agreement (80% within 1 year) than did those from the operculum (60%), sphenoid (59%), or dorsal scute (56%). Likewise, age estimates from pectoral fin rays had the lowest mean coefficient of variation (8.2%) followed by sphenoids (9.9%), opercula (11.3%), and dorsal scutes (11.5%). Only the operculum produced biased estimates between readers. Ages from paired fin rays agreed poorly (36% exact, 30% within 1 year) although no aging bias occurred. The pectoral fin ray is typically used to age shovelnose sturgeon. Because uncertainty about accuracy and precision of age estimates from this structure remains, shovelnose sturgeon management objectives that result from age data should remain conservative.     

Seeking the true time: Exploring otolith chemistry as an age-determination tool

Fish otoliths' chronometric properties make them useful for age and growth rate estimation in fisheries management. For the Eastern Baltic Sea cod stock (Gadus morhua), unclear seasonal growth zones in otoliths have resulted in unreliable age and growth information. Here, a new age estimation method based on seasonal patterns in trace elemental otolith incorporation was tested for the first time and compared with the traditional method of visually counting growth zones, using otoliths from the Baltic and North seas. Various trace elemental ratios, linked to fish metabolic activity (higher in summer) or external environment (migration to colder, deeper habitats with higher salinity in winter), were tested for age estimation based on assessing their seasonal variations in concentration. Mg:Ca and P:Ca, both proxies for growth and metabolic activity, showed greatest seasonality and therefore have the best potential to be used as chemical clocks. Otolith image readability was significantly lower in the Baltic than in the North Sea. The chemical (novel) method had an overall greater precision and percentage agreement among readers (11.2%, 74.0%) than the visual (traditional) method (23.1%, 51.0%). Visual readers generally selected more highly contrasting zones as annuli whereas the chemical readers identified brighter regions within the first two annuli and darker zones thereafter. Visual estimates produced significantly higher, more variable ages than did the chemical ones. Based on the analyses in our study, we suggest that otolith microchemistry is a promising alternative ageing method for fish populations difficult to age, such as the Eastern Baltic cod.     

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.     

Abrupt changes in defence and trophic morphology of the giant threespine stickleback (Gasterosteus sp.) following colonization of a vacant habitat

To evaluate the first stages of adaptation and differentiation following colonization into a vacant and ecologically divergent habitat, 100 melanistic and low‐plated adult threespine stickleback (Gasterosteus aculeatus) exhibiting gigantism in body size and robust spines were transferred in 1993 from a large dystrophic lake (Mayer Lake, Haida Gwaii, British Columbia, Canada) with a diverse vertebrate predator regime, into an adjacent fishless eutrophic pond with macroinvertebrate piscivores. Eight generations were examined as a test of multiple functional hypotheses on the morphological traits that differentiate lake and pond stickleback throughout their distribution. Measurements of 20 defence and trophic traits were made on 275 wild‐caught fish collected from the source and transplant populations over multiple years. There have been significant reductions (males: females) in plate count (8%: 0.5%), lateral plate 2 frequency (46%: 17%), lateral plate 3 height (17%: 18%), pelvic spine length (12%: 6%), dorsal spine length (21%: 16%), gill raker number (5%: 1%), and gill raker length (37%: 43%), and an increase in gill raker spacing (+5%: 0%), jaw length (+5%: +4%), and eye diameter (+8%: +7%). These changes within eight generations represent one‐third of the morphological differences observed between naturally established large lake and pond populations in these archipelago and are all in the direction predicted from the change in habitat. Field samples indicate strong selection in the colonists although lab‐reared individuals implicate an additional role of plasticity for dorsal spine and gill raker lengths, which may contribute to the rapid adaptation into novel and highly divergent selective landscapes. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 107 , 494–509.     

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.     

How old are you—Evaluation of age reading methods for the invasive round goby (Neogobius melanostomus,Pallas 1814)

In the Baltic, the first observation of the round goby (Neogobius melanostomus, Pallas 1814) was made in 1990. Within the past decade the species became invasive and spread rapidly throughout the Baltic Sea. Studies about the fishes potential impacts on resident species promote the need for an increasing knowledge of their basic stock structures such as growth rates, longevity and mortality, which all rely on accurate estimates of age. Former studies on the round goby have used several different age reading techniques. In this study, we compared three standard otolith preparation methods for ageing and present the best procedure for the invasive round goby. The results showed significant differences in age estimates of the same fish between the different preparation methods and between readers. The estimation of the first annulus, the first year, was the most problematic. The overall agreement was lowest when reading the whole otoliths while the best performance was achieved with sectioned and stained preparation method. Depending on method used the growth estimates also differed. The results question comparability between previous studies and highlight the importance of harmonised aging procedures for the round goby for obtaining correct estimates of population parameters such as growth rate, age at maturity, and longevity.     

Age estimates derived from hard parts of swordfish Xiphias gladius from the north-western Mediterranean Sea

Accurate age estimates for fish are critical for properly understanding stock dynamics and health; this is particularly true for larger billfishes. Here we determined the most accurate aging estimation methods for swordfish (Xiphias gladius). We compared age estimates obtained from fin-ray sections, otolith sections, whole otoliths, and vertebrae collected from 87 swordfish off the east coast of Corsica. Age estimates from otolith sections were most consistently estimated across different readers (lowest average percentage error), followed by fin-ray sections, third vertebrae, and whole otoliths. When the age estimates from the otolith sections were compared with the other three age sclerochronological methods, we found the average percentage error to be lowest between the otolith section and fin-ray methods. However, age estimates from fin rays proved most useful for estimating swordfish younger than 6 years, as the fin ray-based age diverged from that of the otolith sections as the swordfish aged. Combining fin ray and otolith section techniques, we estimated the growth parameters of 1–12-year-old females (L_∞ = 259.412, k = 0.113, t₀ = −2.499) and 1–7-year-old males (L_∞ = 175.543, k = 0.202, t₀ = −2.239). We found that females grew significantly faster than males after 3 years and remained larger thereafter. Our calculated growth rates for this region of the north-western Mediterranean Sea were lower than those of the Atlantic, Pacific, and eastern Mediterranean Sea swordfish populations, and similar to growth rates recorded for the western Mediterranean Sea populations. Our study provides critical knowledge on biological-related parameters to serve as a guide for preserving the swordfish population in the Mediterranean Sea.     

Otolith development in wild populations of stickleback: Jones & Hynes method does not apply to most populations

This paper critiques Jones & Hynes (1950) findings by analysing sequential samples of otoliths from three wild populations of Gasterosteus aculeatus from North Uist, Scotland and Nottingham, England. Contrary to Jones & Hynes (1950), but coincident with the finding of later researchers, our results showed that no true translucent ring formed in the otolith of G. aculeatus during their first summer. The first translucent ring was probably starting to be formed by the end of summer and was completed by the end of their first winter. There was no second opaque ring in the otoliths of G. aculeatus before they passed their first winter. The second opaque ring was just starting to appear by early April of the second year in the southern population i.e. Nottingham, but later, by May, in the northern populations i.e. North Uist. Formation of the opaque ring in G. aculeatus mostly occurs in spring and summer, with younger fish starting earlier. In contrast, the formation of translucent rings is mostly during autumn and winter, but can be more widespread through the year, possibly as a result of reproductive investment.     

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.     

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.     

暗纹东方鲀年龄鉴定的研究

用耳石、脊椎骨、鳃盖骨等识别和鉴定暗纹东方鲀(Takifugu obscurusAbe)的年龄,在描述其轮纹特征基础上,比较其在判读暗纹东方鲀年龄和生长特征上的异同与准确性。结果表明,耳石磨片上的轮纹特征明显、清晰且规律性强,是最好的年龄鉴定材料;脊椎骨轮纹也很清晰,但测量时有一定难度,也是较好的年龄鉴定材料;鳃盖骨上年轮较难判定,只能作为辅助材料。本文还对牙齿能否作为年龄鉴定材料进行了观测。       

Evaluating the effectiveness of teeth and dorsal fin spines for non‐lethal age estimation of a tropical reef fish,coral trout Plectropomus leopardus

This study investigated whether teeth and dorsal fin spines could be used as non‐lethal methods of age estimation for a vulnerable and highly valued tropical fisheries species, coral trout Plectropomus leopardus. Age estimation of individuals from 2 to 9 years old revealed that dorsal spines represent an accurate ageing method (90% agreement with otoliths) that was more precise [average per cent error (APE) = 4·1, coefficient of variation (c.v .) = 5·8%] than otoliths (APE = 6·2, c.v . = 8·7%). Of the three methods for age estimation (otoliths, dorsal spines and teeth), spines were the most time and cost efficient. An aquarium‐based study also found that removing a dorsal spine or tooth did not affect survivorship or growth of P. leopardus. No annuli were visible in teeth despite taking transverse and longitudinal sections throughout the tooth and trialling several different laboratory methods. Although teeth may not be suitable for estimating age of P. leopardus, dorsal spines appear to be an acceptably accurate, precise and efficient method for non‐lethal ageing of individuals from 2 to 9 years old in this tropical species.     

Isolation between sympatric anadromous and resident threespine stickleback species in Mud Lake,Alaska

A sympatric pair of anadromous and resident freshwater threespine stickleback species (Gasterosteus aculeatus species complex) occurs in Mud Lake in the Matanuska-Susitna Valley, Alaska. The two forms differ in an array of morphological traits, including traits associated with predator defense (e.g., spine lengths) and trophic ecology (e.g., number of gill rakers). Mud Lake is only the third lake reported to have anadromous stickleback (which have a complete row of lateral plates) coexisting with low-plated resident stickleback in the absence of intermediate partially plated fish. Microhabitat and seasonal isolation appear to contribute to reproductive isolation between the two forms.     

Age and growth of a sub-Antarctic notothenioid, Patagonotothen ramsayi (Regan 1913), from the Falkland Islands

The Falklands’ rockcod Patagonotothen ramsayi was aged successfully using whole and sectioned otoliths. Marginal increment analysis showed that one opaque and one translucent zone were laid down each year. Counting daily rings in juvenile fish and back calculating to their assumed hatch dates validated the first annulus. Readings taken from scales and otoliths showed good agreement with no significant difference between them (P>0.05). Inter- and intra-reader comparisons also showed good agreement. The maximum estimated age was found to be 14 years and the calculated von Bertalanffy growth curve L _T =33.77(1−e ^{−0.25year(t+1.07)}) showed that P. ramsayi is a relatively slow growing fish that attains 5–6 cm L _T in its first year and after which grows approximately 3 cm per year until 4 years. Males seemed to have a slightly lower growth rate but attained a greater maximum size than females. The formation of annuli in the otoliths of P. ramsayi seems to coincide with periods of high reproductive activity with both peaks in GSI and the prevalence of translucent margins occurring in July.