Morphometry and composition of aragonite and vaterite otoliths of deformed laboratory reared juvenile herring from two populations

Vaterite otoliths were sampled from two reared populations (Celtic and Clyde Seas) of juvenile herring Clupea harengus. The crystallography, elemental composition and morphometry were analysed and compared with those of normal aragonite otoliths. The incidence of vaterite otoliths in the juveniles sampled (n = 601) ranged from 7·8% in the Clyde population to 13·9% in the Celtic Sea population, and was 5·5% in the small sample (n = 36) of wild adults examined. In all but one case fish had only one vaterite otolith; the corresponding otolith of the pair was completely aragonite. Although the majority of the juveniles sampled showed craniofacial deformities, there was no link between the skull or jaw malformation and the incidence of vaterite otoliths. All vaterite otoliths had an aragonite inner area, and vaterite deposition began sometime after the age of 90 days. The vaterite otoliths were larger and lighter than their corresponding aragonite partners, and were less dense as a consequence of the vaterite crystal structure. The vaterite areas of the otoliths were depleted in Sr, Na and K. Concentrations of Mn were higher in the vaterite areas. The transition between the aragonite inner areas and the vaterite areas was sharply delineated. Within a small spatial scale (20 μm³) in the vaterite areas, however, there was co‐precipitation of both vaterite and aragonite. The composition of the aragonite cores in the vaterite otoliths was the same as in the cores of the normal aragonite otoliths indicating that the composition of the aragonite cores did not seed the shift to vaterite. Vaterite is less dense than aragonite, yet the concentrations of Ca analysed with wavelength‐dispersive spectrometry (WDS) were the same between the two polymorphs, indicating that Ca concentrations measured with WDS are not a good indicator of hypermineralized zones with high mineral density. The asymmetry in density and size of the otoliths may cause disruptions of hearing and pressure sensitivity for individual fish with one vaterite otolith, however, the presence of vaterite otoliths did not seem to affect the growth of these laboratory reared juvenile herring.     

Significance of otolith calcium carbonate crystal structure diversity to microchemistry studies

Otoliths, calcium carbonate (CaCO₃) ear stones of fish, contain a wealth of information about fish life and environmental history yet the CaCO₃ polymorph form the otolith is made of is a critical, but seldom considered, piece of information during otolith analysis. Otolith trace element chemistry data increasingly informs management decisions, but recent work has shown that CaCO₃ polymorphs—aragonite, vaterite, and calcite—can bear on incorporation of trace elements in a non-trivial way. Most fishes are thought to have otoliths of the aragonite CaCO₃ form, but this construct is potentially outdated with many recent literature reports showing otherwise. Our study used previously unpublished neutron diffraction data and reports from published literature to address three objectives: (1) summarize the relative effects of otolith CaCO₃ polymorphism on otolith microchemistry, (2) summarize reports of otolith polymorphs to gain a better understanding of the extent of non-aragonite otoliths among fishes, (3) outline future research needed to align interpretations of microchemistry with our current understanding of otolith polymorph diversity. We found that while aragonite otoliths are the most common, so are exceptions. For example, the ostensibly rare (among species) CaCO₃ form vaterite was reported in at least some otoliths of 40% of the species surveyed. Our work suggests that examination of the CaCO₃ polymorph composition of otoliths should become more common particularly in studies where results will or may be used to inform management decisions. Future research should work to attribute controls on otolith CaCO₃ polymorph expression using a combination of -omics and material characterization approaches to enrich the life history and environmental information output from otoliths and increase our understanding of the assumptions made in otolith trace element chemistry studies.

     

First record of anomalous otoliths of Menticirrhus americanus in the South Atlantic

Sagitta otoliths are usually formed of calcium carbonate polymorphs as aragonite. The objective of this study was to verify which carbonate polymorph is predominant in the sagitta otolith of Menticirrhus americanus and check whether this pattern remains in otoliths with morphological alterations. Otoliths of M. americanus were obtained from five sites on the southeast‐south coast of Brazil (São Sebastião (SS) 23°45′S–45°24′O, n = 29; Cananéia‐Iguape Estuarine Complex (CI) 25°02′S–47°54′O, n = 30; Paranaguá Estuarine Complex (PEC) 25°28′S–48°20′O, n = 35; Itapoá (IT) 26°07′S–48°36′O, n = 31; Laguna (LA) 28°28′S–48°46′O, n = 13). The characterization of carbonate polymorphs of otoliths was performed through Raman spectroscopy, a photonic and non‐destructive technique that analyzes molecular vibrations induced by laser. We analyzed 138 pairs of M. americanus otoliths, of which eight otoliths from different pairs presented morphological alterations (SS n = 1, CEP n = 5, IT n = 1, LA n = 1). The Raman spectra show that normal otoliths, that is, without morphological alterations, presented only aragonite in their structure. Among the otoliths that presented morphological alterations, the Raman spectra allowed to identify in six otoliths the deposition of aragonite and in only two otoliths the deposition of vaterite (one specimen of the PEC and one of SS).     

High incidence of otolith abnormality in juvenile European flounder Platichthys flesus from a tidal freshwater area

Otolith abnormality has been reported for a wide range of freshwater and marine fish species. In this study, the sagittal otolith morphology and mineralogy of juvenile European flounder, Platichthys flesus, were examined to describe the incidence and types of structural deformities in this species. Juvenile flounder were collected over the years 2013–2015 in the tidal freshwater section of an estuarine nursery (Minho estuary, NW Portugal). Otolith abnormality was found in 43% of the individuals and, in most cases, occurred in both otoliths. Despite an abnormal mineralogy confirmed by scanning electron microscopy, the morphometry and the mineral polymorph composition of abnormal otoliths did not differ from normal ones (i.e. aragonite). This contrasts with other studies where vaterite and/or calcite were found to replace aragonite in abnormal otoliths. Further studies are needed to elucidate whether abrupt salinity changes associated with habitat transitions may have played a role in the abnormal otolith biomineralization in this species.     

Morphology and microchemistry of abnormal otoliths in the ayu, <Emphasis Type="Italic">Plecoglossus altivelis</Emphasis>

The sagittal otolith morphology and microchemistry of reared juvenile ayu, Plecoglossus altivelis, were examined to describe the occurrence and microchemical characteristics of the abnormal otoliths in this species. Juvenile ayu (N = 31) were collected in June 2004 at three different locations, Wakayama, Kumamoto, and Biwa Lake in Japan, where they were being reared in freshwater aquaculture ponds after having been collected in the wild as larvae. Otolith abnormality was found in the sagittae of 26% (N = 8) of the individuals examined, of which five fish had abnormal otoliths only on one side, while the otolith on the other side was normal. Abnormal otoliths were more transparent and crystalline in appearance with irregular shapes compared to normal ones that were more opaque and less irregular. Abnormal otoliths were divided into two types, semi-abnormal (Type 1) with a normal part in the center, and fully-abnormal (Type 2) that were completely crystalline in appearance. The line transects and whole otolith concentration maps showed that the contents of Sr, Na and K were lower in the abnormal otolith regions compared to the normal ones, while those of Ca and S were almost constant in both. The appearance and microchemical properties of the abnormal ayu otoliths were similar to the abnormal otoliths in other species in which vaterite replaces the aragonite. Abnormal formation of otoliths occurred in ayu from Biwa Lake (30%) and Kumamoto (45%), while the Wakayama samples had no abnormality. The microchemical analyses of the normal and abnormal otoliths indicated that some abnormal otoliths had formed before the fish were captured and transferred to the hatchery, so the possible causes of otolith abnormality in ayu are discussed.     

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.     

Analysis of the soluble matrix of vaterite otoliths of juvenile herring (Clupea harengus): do crystalline otoliths have less protein?

Otoliths are calcium carbonate concretions laid down in the inner ear of fish and used in fish age estimation. Otoliths precipitate in the form of aragonite but aberrant precipitation may result in vaterite formation instead of aragonite. Vaterite otoliths are more translucent than aragonite. The quantity of HCl-soluble proteins (SP) was measured in the vaterite otoliths and their aragonite pairs of one year old reared herring Clupea harengus to assess the changes induced by the precipitation of vaterite in the amount of soluble proteins in the otolith. Results showed that vaterite otoliths had as much soluble proteins as their aragonite pairs (p>0.05). Due to the lower density of the vaterite, vaterite otoliths were lighter than their aragonite pairs (p<0.05) which explained that protein concentrations were significantly higher (p<0.05) than in aragonite otoliths. These results indicate that the precipitation of vaterite in otoliths did not affect the inclusion of soluble proteins. Furthermore, they suggest that soluble proteins do not affect the translucent or opaque appearance of otoliths. Differences in translucency may instead be caused by the amounts of insoluble proteins or by differences in the physical properties of proteins. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of the otolith proteins revealed two bands at 50 and 62 kDa in both aragonite and vaterite otoliths suggesting that the precipitation of vaterite in the otolith is not controlled by either of these two proteins present in the otolith.     

Frequency of vateritic otoliths and potential consequences for marine survival in hatchery-reared Atlantic salmon

Otoliths are inner-ear structures of all teleost fish with functional importance for hearing and balance. The otoliths usually consist of aragonite, a polymorph of calcium carbonate, but may also take the form partly or entirely of vaterite, a different polymorph of calcium carbonate. Vateritic otoliths occur sporadically in wild fish, but with a higher frequency in hatchery-reared fish. Abnormal otoliths have direct consequences for the inner-ear functions of fish and may be a symptom of environmental stress. In this study, the authors assess the differences in the frequency of abnormal otoliths and degree of abnormality (% vaterite) for different groups of hatchery-reared Atlantic salmon (Salmo salar) smolt and adults. The groups differed in parental broodstock origin (number of generations in hatchery) and treatment temperature. Smolt from the same groups were also released to complete their ocean migration. The otoliths of the returning and recaptured adults were subsequently extracted to assess the difference in frequency and degree of abnormality between the adults and the smolt from corresponding groups. Return rate varied among groups (0.2%–2.6%). The frequency of vateritic otoliths was high (11.4%–64.4%) and differed among smolt groups. The lowest return rates corresponded with the highest frequency of abnormal otoliths for the groups, suggesting that abnormal otoliths may have negative consequences for marine survival. Furthermore, indications of an effect of fast growth on the formation of abnormal otoliths were found for only one of the experimental groups, and for none of the groups after correcting for Type 1 error. This contradicts previous reports, suggesting rapid growth as the main cause of abnormal otoliths. Adult return rates were generally low, but abnormal otoliths were common, with high coverage (% vaterite).     

Scanning electron microscopic and X-ray diffraction studies of otoconia in the lizard Podarcis s. sicula

Summary The otoliths of embryos and young animals of the lizard Podarcis s. sicula were studied by X-ray diffraction and scanning electron microscopy. Two types of crystal that give different X-ray diffraction patterns were found in the membranous labyrinth of Podarcis. The crystals consist of calcite or aragonite and are easily distinguished by scanning electron microscopy because of their different morphology. The two calcium carbonate crystal forms are not mixed at random but are present in the embryo from the very beginning in specific sites. The endolymphatic sac contains aragonite crystals while the saccule contains calcite crystals adjacent to the wall, in addition to a preponderance of aragonite crystals. The utricle and lagena contain only calcite crystals. The presence of two crystal forms of calcium carbonate in the membranous labyrinth are discussed in terms of differing genetic and functional significance.     

Scanning Electron Microscopy and X-ray Diffraction Studies of the Macular Crystals in the Chondrostean Fish Erpetoichthys calabaricus (Smith)

Morphologic and crystallographic studies of the otoliths of the reed-fish Erpetoichthys calabaricus showed (1) aragonite statoliths with a serrated surface, and (2) two populations of statoconia: one of numerous discoid biconvex crystals of vaterite, the other of pseudohexagonal crystals of aragonite. We suggest that the presence of two calcium carbonate polymorphs in the statoacoustic organs of this archaic fish may have an evolutionary, as well as a systematic and functional significance.     

Exposure to thyroid hormone in ovo affects otolith crystallization in rainbow trout Oncorhynchus mykiss

Calcareous otoliths in the inner ears of fishes are necessary for proper hearing and vestibular function. Sagittal otoliths are usually composed of the calcium carbonate polymorph aragonite but may contain the polymorph vaterite, a phenomenon called otolith crystallization. The causes of otolith crystallization are poorly understood. Thyroid hormone (TH) can influence the chemical microenvironment and structure of the inner ear, suggesting that TH may influence otolith crystallization. The present study examined the effect of exogenous TH treatment on sagittal otolith crystallization and growth in larval and juvenile rainbow trout, Oncorhynchus mykiss. In the first experiment, 110?C179?day-old fish raised from TH-treated oocytes had significantly fewer sagittal otoliths containing the crystalline calcium carbonate polymorph vaterite as compared to untreated fish. Vaterite-containing otoliths were significantly longer than those containing the typical polymorph aragonite, although there was no effect of TH treatment on otolith length. In the second experiment, juveniles immersed in an exogenous solution of TH for 6?weeks had slightly longer otoliths (relative to fish length) than age-matched controls, but this effect was not significant. This juvenile population had a very high percentage (88.3?%) of vaterite sagittae overall and this percentage did not change significantly with treatment, suggesting the switch from aragonite to vaterite occurred prior to inclusion of the fish in the study. These results suggest that early manipulation of TH levels may affect calcium carbonate deposition on the otolith but that later TH exposure is unable to restore typical otolith composition.     

Crystalline otoliths in teleosts: Comparisons between hatchery and wild coho salmon (<Emphasis Type="Italic">Oncorhynchus kisutch</Emphasis>) in the Strait of Georgia

Otoliths, or ‘ear stones’, are calcium carbonate structures found in all vertebrates. In teleosts, they have a number of sensory functions, including hearing. Daily growth increments of these structures have permitted advanced age and population studies of teleosts. Whereas ‘normal’ otoliths are composed of crystals imbedded within a protein matrix as aragonite, a ‘crystalline’ form of calcium carbonate termed vaterite is also found. A review of the otolith literature demonstrates a significant level of understanding of the structure and function of otoliths, but the cause for crystalline otolith structure remains speculative. Pairs of otoliths from hatchery and wild juvenile and adult coho salmon (Oncorhynchus kisutch) were examined visually for determination of otolith microstructure type. The vateritic or crystalline otoliths were found in much higher percentages in juvenile hatchery-reared coho salmon than in juvenile wild coho salmon, supporting previous studies. There did not seem to be any negative impact on size or survival. There was also no correlation between crystalline otoliths and premature maturation in coho males. A preliminary study of adult coho salmon returning to Big Qualicum and Chilliwack hatcheries showed even higher ratios of vateritic otoliths than observed in juveniles.     

Seasonal Stable Isotope Records of Otoliths from Ocean-pen Reared and Wild Cod, Gadus morhua

Otoliths of ocean-pen reared cod, Gadus morhua, provide a unique opportunity to examine the lifetime history of the fish. Here we report 18 analyses of such otoliths on seasonal (winter and summer) stable oxygen and carbon isotope ratios. The calculated isotopic temperatures from otoliths of reared cod were essentially in agreement with the temperature record during rearing, suggesting that temperature is a dominant factor in the precipitation of otolith aragonite. Carbon isotope ratios increased with age and leveled off at 4 years old, presumed to correlate with sexual maturity of cod. As compared with otoliths of wild-caught cod, significant differences were found in isotope variation and the correlation between ¹³C and ¹⁸O. These differences were probably attributable to the different environmental constraints and food supply.     

The growth ofOreochromis andersonii (Pisces: Cichlidae) from the Okavango Delta,Botswana, and a comparison of the scale and otolith methods of ageing

Synopsis Otoliths and scales were used for age and growth determination ofOreochromis andersonii from the Okavango Delta, Botswana. Marginal increment analysis showed that an annulus was formed in both the scales and otoliths during the dry summer period. Using scales, the growth ofO. andersonii was described by L_t = 285.27(1-e^{-0.26(t+2.02)}) mm SL and using otoliths by the equation L_t = 267.48(1-e^{-0.25(t+2.18)}) mm SL. Maximum age estimates of 10 years using scales and 13 years using otoliths were obtained and the growth curves were significantly different (p < 0.01). Age estimation using scales tended to over-emphasise growth inO. andersonii resulting in larger predicted lengths-at-age. For this reason, otoliths are considered to be more reliable and suitable than scales in determining the age and growth of this species.     

Strontium:cakium concentration ratios in otoliths of herring larvae as indicators of environmental histories

Synopsis Elemental analyses, using wave-length dispersive electron microprobe techniques on otoliths from reared Atlantic herring larvae, Clupea harengus, showed trace quantities of strontium relative to that of calcium, and an inverse relationship between Sr/Ca concentration ratios and rearing temperature. These data are consistent with those for coral aragonite, in that there appears to be an inverse temperature effect on physiological incorporation of strontium in the otolith aragonite. Our determinations of Sr/Ca concentration ratios of lab-reared herring larvae showed that the deposition of strontium relative to calcium and the rearing temperature were related, where: T (° C) = −2.955 [Sr/Ca] × 1000 ± 19.172. This principle thus makes it possible to use Sr/Ca concentration ratios in fish otoliths to delineate past temperatures experienced by an individual. Further, combining electron microprobe analyses with scanning electron microscope (SEM) examinations of daily increments in the same otolith makes it possible to reconstruct the temperature history for an individual fish on a time scale of days. An example of the application of the technique to an approximately six-month-old field-caught herring larva is given, and the limitations of the technique are discussed.     

Biomarkers from individual carbonate phases of an Oligocene cold‐seep deposit,Washington State,USA

An Oligocene cold‐seep limestone (Lincoln Creek Formation, Washington State, USA) was studied for its lipid biomarker inventory. Biomarker analysis on minute amounts of sample (tens of mg) and complementary ¹³C_carbonate analyses allowed us to link biogeochemical processes with individual, closely intertwined carbonate phases. The ancient seep deposit exhibits four major carbonate phases, according to the paragenetic sequence of (I) micrite, (II) yellow aragonite, (III) clear aragonite and (IV) equant calcite spar. For the micrite, varying but significant amounts of archaea‐derived isoprenoids clearly indicate that the precipitation of this phase was induced by the microbial anaerobic oxidation of methane (AOM). However, water column‐derived lipids present in this carbonate phase reflect the incorporation of organic matter from background sediment cemented by authigenic micrite. Yellow aragonite made up only a minor rock component (<10% vol.), but contained a major portion of lipid biomarkers indicative of AOM. Along with low δ¹³C_carbonate values (less than ?30‰ Pee Dee Belemnite), this points to an intimate spatial association of AOM consortia with the precipitation of yellow aragonite. Clear aragonite showed similar δ¹³C_carbonate values but much lower, if any, contents of AOM biomarkers. This suggests that AOM‐derived carbonate ions diffused over a greater distance to the site of precipitation compared with yellow aragonite. The latest phase, equant calcite spar, did not yield appreciable biomarkers, but showed a notable ¹³C_carbonate‐enrichment that is most likely caused by methanogenesis that prevailed in the sediments after AOM activity had ceased. A comparison of the ancient seep carbonates with modern counterparts from Hydrate Ridge (offshore Oregon, USA) revealed a remarkable coincidence of the respective mineral phases and their biomarker patterns. This suggests that the mechanisms of carbonate formation and the associated biogeochemical processes remained unchanged over geological times. □Biomarkers, carbonates, cold seeps, Hydrate Ridge, isotopes, Oligocene, Washington.     

Age,growth and reproduction in creole perch (Percichthys trucha) in the Negro River,Argentinean Patagonia

Age, growth and reproductive characteristics of creole perch, Percichthys trucha, were investigated in the Negro River, southern Argentina from samples collected seasonally, December 1994–December 1995. Age was estimated via scale and whole otolith reading methods. Total length (n = 413) ranged from 103 to 432 mm, and weight from 12 to 1042 g. Significant differences between the length‐weight relationships of males and females were detected (P < 0.05). Isometric growth was observed in juveniles and males, whereas total population and females exhibited positive allometric growth. The marking pattern in scales and otoliths followed an annual rhythm, with the formation of only one annulus in scales and only one hyaline band in otoliths during autumn‐winter. The oldest males were 5 years old whereas maximum age in females was 12 years from scales and 15 years from otoliths. Because scales were found to underestimate age in individuals older than 4 years, otoliths were considered to be the best structures for creole perch age determination. Gompertz growth parameters based on otolith data were L∞: 428.0 mm, k = 0.46 and t₀ = 0.43 for total population (r = 0.90), L∞: 410.7 mm, k = 0.42 and t₀ = 0.46 for males (r = 0.91), and L∞: 434.1 mm, and k = 0.49 and t₀ = 0.43 for females (r = 0.91). Lengths at first maturity (TL₅₀) were 260 and 241 mm in males and females, respectively, both of which corresponded to ages between 1 and 2 years. Macroscopic gonad inspection and the high percentage of juveniles captured during summer indicated that spawning begins at the end of spring.     

Discriminant analysis as a tool to identify catfish (Ariidae) species of the excavated archaeological otoliths

Catfish otoliths excavated from two archaeological sites in Kuwait, Sabiyah (ca. 7000 Years Before Present) and Al-Khidr, ca. 4000 YBP, were compared with those of Kuwait’s modern catfish. Otoliths from Kuwait’s four species of catfish, Netuma bilineata, N. thalassina, Plicofollis dussumieri, and P. tenuispinis were collected after recording total length and weight. Data recorded for both ancient and modern otoliths, including annual ring (age), weight, length and four otolith radii from transverse sections, were subject to discriminant analysis to differentiate among species and develop classification functions for otoliths. Comparisons of the results from the ancient and modern otoliths showed that most of the excavated otoliths (78% from Sabiyah and 100% from Al-Khidr) belong to the two presently dominate species N. bilineata and P. tenuispinis, indicating that ichthyofauna of Kuwait Bay may not have changed much in the past 7000 years.     

The use of sectioned otoliths to age barramundi (Lates calcarifer) (Bloch, 1790) [Centropomidae]

The relationship between the number of rings present in sagittal otoliths and the age of barramundi, Lates calcarifer (Bloch, 1790) [Centropomidae], was investigated by examining cross sectioned otoliths of 37 tagged fish of known age between 1 and 5 years from the Johnstone River, north Queensland. Concentric rings were clearly visible in all otolith sections and were validated as annual marks. The technique was then used to estimate the age and calculate von Bertalanffy growth parameters for 70 barramundi from the Fitzroy River, central Queensland. Growth appeared to be rapid but variable in the first year; the von Bertalanffy growth parameters for length versus age were L =690 mm, K=0.53, t ₀= 0.003 years. October 1 was designated as the birth date. Whole otolith length, width and thickness were also approximated well by the von Bertalanffy equation. We suggest that examination of otoliths is a useful technique for ageing barramundi but note that further validation of the ageing method is still needed for fish older than six years.     

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.