Growth acceleration,behaviour and otolith check marks associated with sex change in the wrasse <Emphasis Type="Italic">Halichoeres miniatus</Emphasis>

In protogynous sex-changing fishes, females are expected to compete for the opportunity to change sex following the loss of a dominant male and may exhibit growth and behavioural traits that help them maintain their dominant status after sex change. A male removal experiment was used to examine changes in female growth and behaviour associated with sex change in the haremic wrasse Halichoeres miniatus and to test whether any changes in growth associated with sex change were recorded in otolith microstructure. Dominant females began displaying male-characteristic behaviour almost immediately after the harem male was removed. The frequency of interactions between females increased following male removal. In contrast, feeding frequency of females decreased. The largest one to three females in each social group changed sex following male removal and exhibited an increase in growth associated with sex change. Sex changers grew more than twice as fast as non-sex changers during the experimental period. This growth acceleration may enable new sex-changed males to rapidly reach a size where they can defend the remaining harem from other males. An optical discontinuity (check mark) was present in the otoliths of sex-changed fish, and otolith accretion rate increased significantly after the check mark, corresponding with the increased growth rate of sex-changing females. Wild caught males, but not females, exhibited an analogous check mark in their otoliths and similar increases in otolith increment widths after the check. This indicates that an increase in growth rate is a regular feature of sex-change dynamics of H. miniatus. Communicated by Environment Editor Prof. Rob van Woesik     

Evidence of perturbations induced by reproduction on somatic growth and microincrement deposition in Oreochromis niloticus otoliths

The direct effects of reproduction (energy expenditure for gametogenesis) were studied in gonadectomized Oreochromis niloticus females, incapable of reproducing. The indirect effects of reproduction were studied in intact females that had either mouth brooded their eggs or not. In gonadectomized females the slowing of growth observed in the marginal zone of their otoliths coincided with the time of gonad regeneration. The responses were very varied in the group of intact females. In these females, there was less otolith growth and more checks were observed on the otoliths. The results showed that reproduction influenced the growth of fish and their otoliths leading to a significant underestimate of the number of primary increments during this period of life.     

Estimating the timing of growth rings in Atlantic cod otoliths using stable oxygen isotopes

A technique involving micro‐scale sampling of otolith carbonate and analyses of stable oxygen isotope composition was used to relate the zone appearance of the otolith to the seasonal temperature cycle. Otolith opacity could then be related to the timing of zone formation. Otoliths from two groups of Atlantic cod Gadus morhua held under known temperature conditions over a period of 4 and 6 years were examined. The otolith translucency followed the same pattern as the estimated temperature (from otolith δ¹⁸O values) in the yearly increments three and four, meaning that the translucent zones were deposited at the seasonal highest temperature in late summer and early autumn. The relative light intensities of otolith yearly increments five and six of older fish (deposited in the same years), however, were not significantly correlated to the estimated temperatures since increased otolith translucency also occurred at low temperatures. This might have been caused by stress in connection with gonad development or starvation during the spawning period. The results showed that this method of coupling otolith opacity and stable oxygen isotope composition can be used to estimate the timing of zone formations in otoliths.     

An evaluation of the otolith characteristics of Conger conger during metamorphosis

A sample of 20 metamorphosing conger eel Conger conger leptocephali were collected from the Minho River, Portugal, in February 1999 and their sagittal otoliths were analysed by scanning electron microscopy. Four different etching agents were applied along both sagittal and frontal sections during otolith preparation to examine the microstructural growth in this species. Otolith growth increments were visible throughout the increment countable zone using all four treatments, but a permanent peripheral diffuse zone, where the daily increments were unclear, appeared on all otoliths, preventing accurate age estimation. To understand more about the nature of the diffuse zone, otoliths of 10 other metamorphosing leptocephali reared in aquaria were marked by immersion in tetracycline hydrochloride. The distance between the fluorescent marks and otolith edge, measured over a fixed period of time, was used to estimate the otolith growth rate. The application of this technique led to an anomalously high estimated otolith growth rate, probably as a result of the capture, marking and handling stress.     

Increased H-Y antigen levels associated with behaviorally induced, female-to-male sex reversal in a coral-reef fish

It has been proposed that H-Y antigen is the synthetic product of sex-determining genes, and that H-Y antigen controls ontogenetic differentiation of the heterogametic sex throughout vertebrates. The coral-reef fish Anthias squamipinnis is a protogynous hermaphrodite in which all individuals mature initially as females. Males result when adult females change sex as a consequence of alterations in behavioral interactions within social groups. Three assay methods were used to measure H-Y antigen levels in the spleens, gonads, and epidermal tissue of 16 adult females and in 16 males that had been induced to change sex from a prior female phase by the removal of a pre-existing male from each of 16 social groups. In 15 male-female pairs, the H-Y antigen levels were higher in male than in female spleen, gonad, and epidermis tissues. The precise temporal relationship between the onset of sex change and the increase in the H-Y antigen level was not examined. If, as we strongly suspect, the temporal relationship proves to be close, the inference will be that the behavioral cues inducing sex change also influence the synthetic activity of genes controlling H-Y antigen production.     

The effect of otolith storage methods on the concentrations of elements detected by laser-ablation ICPMS

Otoliths of male (1+ year old) Terubok, tropical shad Tenualosa toli from the Lupar River, Sarawak, were treated in different ways to assess the effect of post-capture storage on element concentrations in the otolith core and edge detected by laser-ablation inductively coupled plasma mass spectrometry (ICPMS). Five treatments were compared: rapid removal and dry storage; fish kept 5 h after capture then otoliths removed; rapid removal and stored in alcohol; fish rozen for 24 h then otoliths removed; and storage of intact head in alcohol for several weeks. Seven elements were detected at the core and edge of otoliths (Li, Na, Mg, Mn, Co, Sr, Ba). Of these, Na, Mg, Co and Ba differed among treatments, being highest in treatments where the otolith remained in the head. Variation in element measurements due to treatment effects was similar to within-otolith variation for the other elements and was least variable in the fresh treatment. The variation in concentration between the core and edge was more significant than other sources of variation (up to 100 times) for some elements. These differences were probably due to physiological factors (ontogenetic) or to yearly changes in water chemistry. The results indicate that the method of otolith storage, especially freezing whole fish, can have a small, but measurable, effect on the concentrations of Na, Mg, Co and Ba.     

Three-dimensional imaging of walleye pollock otoliths: reconstruction from serial sections and fluorescent laser cytometry

Two techniques have been developed to examine the three-dimensional internal structure of otoliths. In the first, otoliths were sectioned serially, images were digitized, and the otolith was reconstructed as a computer model. In the second method growth increments were marked in vivo during their formation by immersing the fish in a fluorescent dye, and then the internal structure of the otolith visualized using laser cytometry. The results are useful for evaluating the potential for bias in otolith measurements and for determining the sectional plane with the least bias.     

Enigmatic ear stones: what we know about the functional role and evolution of fish otoliths

Otoliths in bony fishes play an important role in the senses of balance and hearing. Otolith mass and shape are, among others, likely to be decisive factors influencing otolith motion and thus ear functioning. Yet our knowledge of how exactly these factors influence otolith motion is incomplete. In addition, experimental studies directly investigating the function of otoliths in the inner ear are scarce and yield partly conflicting results. Herein, we discuss questions and hypotheses on how otolith mass and shape, and the relationship between the sensory epithelium and overlying otolith, influence otolith motion. We discuss (i) the state‐of‐the‐art knowledge regarding otolith function, (ii) gaps in knowledge that remain to be filled, and (iii) future approaches that may improve our understanding of the role of otoliths in ear functioning. We further link these functional questions to the evolution of solid teleost otoliths instead of numerous tiny otoconia as found in most other vertebrates. Until now, the selective forces and/or constraints driving the evolution of solid calcareous otoliths and their diversity in shape in teleosts are largely unknown. Based on a data set on the structure of otoliths and otoconia in more than 160 species covering the main vertebrate groups, we present a hypothetical framework for teleost otolith evolution. We suggest that the advent of solid otoliths may have initially been a selectively neutral ‘by‐product’ of other key innovations during teleost evolution. The teleost‐specific genome duplication event may have paved the way for diversification in otolith shape. Otolith shapes may have evolved along with the considerable diversity of, and improvements in, auditory abilities in teleost fishes. However, phenotypic plasticity may also play an important role in the creation of different otolith types, and different portions of the otolith may show different degrees of phenotypic plasticity. Future studies should thus adopt a phylogenetic perspective and apply comparative and methodologically integrative approaches, including fossil otoliths, when investigating otoconia/otolith evolution and their function in the inner ear.     

Influence of water temperature and feeding regime on otolith growth in Anguilla japonica glass eels and elvers: does otolith growth cease at low temperatures?

The influences of water temperature and feeding regime on otolith growth in Anguilla japonica glass eels and elvers were investigated using individuals reared at 5, 10, 15, 20, 25 and 30° C and in fed or unfed conditions at salinity 32 after their otoliths were marked with alizarin complexone (ALC). To eliminate the difficulty of observing the edges of otoliths with optical (OM) or scanning electron (SEM) microscopes, three to 10 individuals were sampled from each tank at 10, 20 and 30 days during the experiment and reared for an additional 10 days at 25° C after their otoliths were marked a second time. Otolith growth and the number of increments were measured using both OM and SEM. Most A. japonica commenced feeding after 10 days at 20–30° C or after 20 days at 15° C, but no feeding occurred at 5 and 10° C. No otolith growth occurred at 5 and 10° C except in two individuals with minimal increment deposition at 10° C. Otolith growth was proportional to water temperature within 15–25° C and not different between 25 and 30° C. At 15, 25 and 30° C, the mean otolith growth rate in fed conditions was higher than in unfed conditions. The number of increments per day was significantly different among water temperatures (0·00–0·01 day⁻¹ at 5 and 10° C, 0·43–0·48 day⁻¹ at 15° C and 0·94–1·07 day⁻¹ at 20–30° C). These results indicated that otolith growth in A. japonica glass eels and elvers was affected by temperature and ceased at ≤10° C under experimental conditions. Hence, future studies analysing the otoliths of wild-caught A. japonica glass eels and elvers need to carefully consider the water temperatures potentially experienced by the juveniles in the wild.     

Sexual selection explains sex-specific growth plasticity and positive allometry for sexual size dimorphism in a reef fish

In 1950, Rensch noted that in clades where males are the larger sex, sexual size dimorphism (SSD) tends to be more pronounced in larger species. This fundamental allometric relationship is now known as ‘Rensch''s rule’. While most researchers attribute Rensch''s rule to sexual selection for male size, experimental evidence is lacking. Here, we suggest that ultimate hypotheses for Rensch''s rule should also apply to groups of individuals and that individual trait plasticity can be used to test those hypotheses experimentally. Specifically, we show that in the sex-changing fish Parapercis cylindrica, larger males have larger harems with larger females, and that SSD increases with harem size. Thus, sexual selection for male body size is the ultimate cause of sexual size allometry. In addition, we experimentally illustrate a positive relationship between polygyny potential and individual growth rate during sex change from female to male. Thus, sexual selection is the ultimate cause of variation in growth rate, and variation in growth rate is the proximate cause of sexual size allometry. Taken together, our results provide compelling evidence in support of the sexual selection hypothesis for Rensch''s rule and highlight the potential importance of individual growth modification in the shaping of morphological patterns in Nature.     

Sexual dimorphism in the otolith shape of shi drum,Umbrina cirrosa (L.), in the eastern Mediterranean Sea: Fish size–otolith size relationships

Little is known about possible differences in sagitta otolith size and shape between sexes of the shi drum, Umbrina cirrosa, and relationships between their body and otolith size. Thus, this study aimed to fill this knowledge gap via examination of 414 sagittal otoliths from 108 male (total length 13.8–26.8 cm) and 99 female (13.5–26.7 cm) U. cirrosa caught between May 2017 and April 2018 in gillnets set at a depth of ~15 m in Mersin Bay, Eastern Mediterranean Sea. No statistical differences were observed between the shape indices of the left-sided and right-sided sagitta. However, there were significant differences in the size and shape of otoliths between males and females. The slopes of allometric power functions from otolith width × fish sizes gave significant differences between males and females (ANCOVA, P < 0.05). The relationship for length × weight of otoliths from both males and females showed isometric growth, whereas the relationship of otolith width × otolith weight showed positive allometry. Negative allometric growth was observed for the relationship otolith length × otolith width. In summary, this study revealed the presence of sexual dimorphism in the otolith shape of U. cirrosa, and the data on regression relationships of fish-otolith sizes can be used to estimate fish size from U. cirrosa otolith sizes.     

Onset of maturity and cohort composition at spawning of Baltic sprat Sprattus sprattus on the basis of otolith macrostructure analysis

Data on catch per unit effort (CPUE) from the Danish commercial fishery showed dense aggregations of sprat Sprattus sprattus born in 1997 in the main spawning area of the Baltic Sea during May 1999. After the analysis of their otolith macrostructure, these fish were found to have a shorter distance to the beginning of the first winter zone compared to fish from the same cohort caught in other months. At the same time, the number of fish with large otoliths at the period of winter zone formation tended to decrease or even disappear. The otolith size at winter zone formation ( O _W) was found to be a good predictor of sprat total length ( L _T) and mass ( M ) in age group 0 years. Changes in the otolith size distribution over time were related to variations in cohort composition depending on the L _T and M attained during the juvenile stage. Examination of the gonads suggested two life history patterns: 1) individuals achieving a larger L _T and M during the first growing season may contribute at age 1 years to the spawning stock and the fishery, whereas 2) individuals attaining smaller L _T and M contribute in larger amounts to both the spawning stock and the fishery later, when they are at age 2 years. Moreover, changes in the relationship between O _W and L _T and M were related to differences in growth trajectories for sprat born during the same spawning season. Smaller sprat as group 0 years continued growing during the second growing season delaying maturity compared to larger individuals from the same cohort.     

Age and growth of the round goby Neogobius melanostomus in the Gulf of Gdańsk several years after invasion. Is the Baltic Sea a new Promised Land?

The ages of 8 to 23·5 cm total length (L(T)) round goby Neogobius melanostomus collected monthly during 2006 and 2007 in the Gulf of Gdańsk (Baltic Sea) ranged from 2 to 6 years, with age class 4+ years dominant. Males were larger at age than females. The fastest growth occurred in the first 2 years of life in both sexes. Females were heavier at a given L(T) than males, but only for fish > c. 15 cm. A strong relationship between N. melanostomus otolith size and fish size was found, with no difference between males and females, and a significant relationship between fish growth rate and otolith growth rate, which enabled backcalculation of growth rates. Marginal increment width analysis confirmed the periodicity of annual ring formation in otoliths and showed that the most intense opaque zone formation occurs in July to August, while hyaline zone formation starts as early as September to October. It was concluded that the N. melanostomus that have colonized the southern Baltic Sea exhibit the largest size and longest life span ever recorded for this species.     

Age validation and growth of three commercially important hemiramphids in south-eastern Australia

The method of using sectioned otoliths to estimate age in three species of garfishes (family Hemiramphidae) was validated by: (1) staining fishes with the vital stain alizarin complexone (ALC) and periodically examining their otolith growth, and (2) marginal increment analyses. Staining fishes with ALC indicated that opaque zones were formed during winter and spring, but did not become visible on the otolith edge until late spring and summer. Hyporhamphus australis were found to be similar to the hemiramphids of the Atlantic in having fast growth rates and a maximum observed age of 4+ years old. Hyporhamphus regularis ardelio and Arrhamphus sclerolepis krefftii were found to be more similar to the southern sea garfish, Hyporhamphus melanochir , in being moderately long-lived, with maximum observed ages of 7+ years old for both species. Females grew faster and attained greater fork lengths than males for each species. Sectioned otoliths showed large variation in the appearance of opaque zones between the three species studied, with those from the wide-ranging, oceanic H. australis appearing inconsistent and diffuse when compared to the estuarine H. r. ardelio and A. s . krefftii . This variation was also apparent from fishes kept in aquaria, suggesting that the appearance of opaque zones in otoliths of these species is largely influenced by physiology, rather than by environmental conditions.     

Do otolith annular structures correspond to the first freshwater entry for yellow European eels Anguilla anguilla in the Baltic countries?

To examine the relationship between freshwater entry and otolith annular structures, a total of 113 naturally recruited European eels Anguilla anguilla from Lithuania and Latvia that entered fresh water at least once were collected. In some individuals (8·3–11·3%), the first freshwater entry coincided with a dark check that was distinctly different from neighbouring annuli. In most individuals (81·7–84·9%), the first freshwater entry occurred on rings and increments indistinguishable from other annuli. For the remaining individuals (3·8–10%), the first freshwater entry did not correspond to any otolith ring, band or annulus. According to recent evidence, the observed high correspondence between the first freshwater entry and otolith annuli was more likely due to the movement into fresh water during winter when the annulus was deposited, rather than stress resulting from habitat change. Consequently, the age estimation based on otoliths might be less influenced by this habitat change during the yellow eel stage.     

Growth and morphological development of sagittal otoliths of larval and early juvenile Trachurus japonicus

The daily periodicity of growth increment formation in sagittal otoliths of jack mackerel Trachurus japonicus was validated by marking otoliths with alizarin complexone (ALC). Analysis of otoliths of known‐age juveniles confirmed that the first increment formed on day 3 after hatching, and was associated with first feeding. A total of 198 specimens, ranging from 2·6 to 49·2 mm in body length (notochord length or standard length) and from 7 to 78 days in age, were collected in the East China Sea and Tosa Bay, and used to examine the association between otolith morphological development and ontogenetic development. The relationship between body length ( L ) and otolith radius ( R ) was significantly described by the linear function L  = 2·65 + 0·0425 R ( n  = 198, r ² = 0·99, P  < 0·001), indicating that somatic growth history can be reconstructed from otolith growth patterns. The otolith was primarily spherical in the preflexion larval stage, and became elongated with notochord flexion. The first secondary primordium formed at c . 25 days, during the middle postflexion stage, and was associated with metamorphosis. By c . 42 days the sagittal otolith was adult‐like in morphology, with the primary growth zone enclosed by the marginal growth zone, except in the anterior rostrum area. Thus age, growth and developmental stages were recorded in sagittal otoliths during the larval and early juvenile stages of jack mackerel.     

THE COSTS OF CHANGING SEX AND THE ONTOGENY OF MALES UNDER CONTEST COMPETITION FOR MATES

In its simplest form, the size-advantage hypothesis predicts that individuals should change sex in order to increase their reproductive success. In terms of lifetime expectations, this must be true for the hypothesis to hold. However, as we review here, some loss of reproductive success may occur immediately after sex change. Unavoidable costs (e.g., those resulting from a restructuring of the gonad) have not been adequately distinguished from adaptive allocations of resources which diminish current reproduction in favor of large increases in future mating success. This strategy can become particularly important for species in which a few males monopolize matings. To illustrate this idea, we describe the changes in mating frequency as mature females become sexually active males in three species of protogynous wrasses. In one species, a male defends a permanent, all-purpose territory composed of up to 12 females. When he is removed, a single female changes sex and successfully completes mating sequences with all females in the territory within an average of 5.6 days. This duration roughly corresponds to the time required for functional transformation of gonads; thus, individuals in this species suffer few reproductive losses as a result of changing sex. The largest males in two other species mate with an average of 25 to 50 females per day, but only by successfully defending reproductive territories. In one of those species, individuals that changed sex mated infrequently over a two-year period after sexual transformation and, by the end of the study, were still well below the average size of males that consistently obtained territories. Sex-changed individuals in the other species had very low reproductive success for up to 45% of the maximum lifespan as a male. It is improbable that the substantial cost of changing sex in the latter two species results from gonad restructuring or from mistakes due to imprecise cues for sex change. Instead, the cost appears to represent an investment in growth rather than current reproduction as a means of rapidly attaining a size advantage when individuals face intense competition for extraordinarily successful mating territories.     

Historical aspects of protandrous sex change in the anemonefish Amphiprion melanopus (Pomacentridae, Teleostei)

Gonadal structure and cellular composition were examined in juveniles, males and females of the protandric hermaphrodite, Amphiprion melanopus. Functional sex change was experimentally induced in the field and gonad structure was histologically examined both qualitatively and quantitatively at 10, 20, 30 and 45 days after its initiation. Juvenile gonads consist primarily of immature ovarian tissue. Functional male gonads are ovotestes with co-existing mature spermatogenic tissue and immature ovarian tissue, while females possess only ovarian tissue. The initiation of sex change is marked by a rapid maturation of spermatogenic tissue and proliferation of putative oogonia. Gonads were essentially female by 20 days into sex change, but evidence of mature female function (marked by the initiation of vitellogenesis) was not observed until 45 days. Considerable variation between individuals was seen in quantitative measures of gonadal change in the early stages of sex change, but not in later stages. Progress in sex change as indicated by histological indicators was, however, consistent within stages. Duet systems for gamete transport changed from the male to the female form after all male tissue had been replaced.     

The periodicity of primary increment formation in the otoliths of Arctic charr Salvelinus alpinus (L.)

The role of environmental factors in the regulation of sub-annual increment formation in fish otoliths appears to differ markedly between species. To examine the periodicity of primary increment formation in the otoliths of O + Arctic charr, Salvelinus alpinus (L.), and the effects of temperature, and photoperiod on their formation, fish were held under controlled environmental conditions. Primary growth increments were found in the otoliths of fish held at constant temperature (18° C) and at ambient temperature [fluctuating with a circadian and circannual rhythm (4–18° C)]. Consistent and significant disruptions in increment formation occurred however, in experimental groups subjected to rapid change from ambient photoperiod to a 6L: 6D photo-period for 96 h. Disruptions in increment formation were also observed immediately following transportation of fish between holding facilities and following disease treatment. The number of otolith increments formed in fish held on an ambient photoperiod regime, correlated closely with time elapsed in days since checkmark formation ( r = 0.989, P ≤0.001) in fish sampled sequentially over a period of 10 to 105 days. Thus we demonstrate that under conditions of ambient photoperiod, primary increments are formed daily.     

The differentiation of Stegastes partitus populations using lapillar and sagittal otolith chemistry

The comparison of elemental concentrations of sagittal and lapillar otoliths from the same individuals of Stegastes partitus indicated significant differences for several elements. Sagittal otoliths were superior at differentiating individuals, yet the differentiation of individuals was further improved when the elemental concentrations of both otolith types were used in the same analysis.