Metazoan parasites as biological tags for stock discrimination of whitemouth croaker Micropogonias furnieri

Examination of 248 adult specimens of whitemouth croaker Micropogonias furnieri from five localities along the Brazilian coast revealed 8735 parasites belonging to 41 metazoan species. Samples from Ceará to Bahia and Rio de Janeiro to Santa Catarina showed a high level of correct assignation (92 and 87%, respectively) and cross assignation (i.e. almost all specimens misidentified in Ceará were assigned to Bahia and almost all specimens misidentified in Bahia were classified as Ceará), so samples were pooled in the northern and south‐eastern samples, and Rio Grande do Sul was considered a southern area. Eight parasite species were characteristic of the northern localities, five species were found just in the area associated with south‐eastern localities and two species were characteristic of the southern area providing first evidence of stock discreteness. The multivariate discriminant analysis successfully discriminated three groups of localities associated with three stocks of M. furnieri in Brazil: a northern stock associated with Ceará and Bahia, a south‐eastern stock related to Rio de Janeiro and Santa Catarina and a southern stock in the area of Rio Grande do Sul, which could be considered as the northern limit of the stock associated with the Common Fishing Zone of Uruguay and Argentina.     

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.     

Ontogenetic and environmental effects on otolith shape variability in three Mediterranean European eel (Anguilla anguilla, L.) local stocks

Otolith morphology is an efficient tool for the discrimination of fish stocks, populations and species when comparative genetic data are not available. Currently, the relationship between environmental factors and otolith shape is poorly characterized for the European eel (Anguilla anguilla), a highly migratory catadromous species constituting a single, randomly mating stock. The present study analyses the differences in otolith morphology between three Mediterranean eel local stocks from different environmental contexts (i.e. two brackish lagoons and one river). The relationship between otolith shape and otolith size was studied by means of Elliptic Fourier analysis and multivariate statistics. Otolith profile was digitally acquired and Cartesian coordinates were extracted. Partial Least Square (PLS) analysis pointed to continuous allometric growth in size and shape in otoliths from all three sites. In the three environments, shape variations occurred during growth as indicated by the presence of a significant and positive relationship between otolith size and the first PLS latent vector (i.e. which bears most of the information regarding otolith outline). Differences between smaller and larger sized otoliths were investigated using PLS Discriminant Analysis (PLSDA) and cluster analysis. Results indicate that otolith shape is highly uniform at smaller than at larger sizes. These shape differences apparently overlap the initial differentiation of the small otolith outlines acquired by eels during the growing phase as elvers in the marine environment. Data were discussed considering that the physical and chemical habitat variability in brackish lagoons and river could underlie a marked change in otolith shape during the animals' growth.     

Otolith size and location in digestive tracts of northern fur seals (Callorhinus ursinus): Implications for dietary interpretations

Walleye pollock (Theragra chalcogramma) otoliths (n= 2,706) recovered from stomachs, small intestines, and colons of 43 northern fur seals (Callorhinus ursinus) were evaluated for size and wear by location in the digestive tract. Pollock fork length was regressed on otolith length after correction for erosion, and age was estimated from the calculated body size. Age‐1+ pollock otoliths (≥6.3‐mm length) were concentrated in stomachs while age‐0 otoliths (≤6.2‐mm length) were concentrated in colons. Less than 10% of otoliths were found in the small intestines. Pollock age decreased with progression along seal gastrointestinal tracts. Otolith quality increased along gastrointestinal tracts in numbers ≥20, which was typical of age‐0 otoliths recovered from colons. Otolith distribution by age and quality along gastrointestinal tracts suggests that small (≤12 cm) schooling prey are consumed in large volume and passed as a bolus rapidly through the digestive tract before significant erosion of bony remains occurs; while larger prey are eaten in smaller volume and subjected to otolith erosion due to longer retention in the stomach. Our results illustrate the importance of multiple sampling strategies to comprehensively represent prey size in pinniped diet.     

Validation of daily otolith increments in larval and juvenile Chinese sucker, <Emphasis Type="Italic">Myxocyprinus asiaticus</Emphasis>

Otolith development was observed and the formation of daily growth increments in otoliths of Chinese sucker, Myxocyprinus asiaticus, was validated by monitoring known-age larvae and juveniles in the laboratory from 2003 to 2005. Otolith shape changed with larval and juvenile development, and there was an exponential relationship until a body length of 16 mm or so, and a linear relationship after a body length of 16 mm between otolith size and fish size. The first increment was identified in larvae 1 day after hatching. The regressed equations between daily age (D) and increment number in otoliths (N) were N = −0.64 + 0.96D in lapillus, and N = −0.31 + 0.98D in sagitta. The slopes were not significantly different than 1.0. This demonstrated that otolith increments in this species were formed daily and can be used for daily age determination.     

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.     

Factors affecting morphological development of the sagittal otolith in juvenile and adult small yellow croaker (Larimichthys polyactis Bleeker, 1877)

The morphology and morphometrics of the sagittal otoliths of small yellow croaker (Larimichthys polyactis) from the southern Yellow Sea were investigated. Study objectives were to evaluate the shape variability and morphometric variables of sagittae of juveniles and adults as related to developmental changes and habit shift. A total of 152 fish were sampled from April to June of 2012 and 2013, along the coastal waters of the Lüsi spawning ground in the southern Yellow Sea. Changes in otolith shapes from the juvenile to the adult were presented with the rim development through the entire‐lobed‐entire transition and with the curvature of the cauda toward the ventral margin. The otolith elongation in the juvenile stage occurred at 10–20 mm standard length (SL) and was likely associated with the formation of otolith accessory growth centers from larvae to juvenile. The L. polyactis sagittal otoliths acquired their definitive shape at 130 mm SL maturity. Ontogeny on otolith shape might be related, for example, to the factors of metamorphic development, feeding habitat, and ambient water salinity, which varied throughout the growth of L. polyactis.     

Stock discrimination of the Japanese Spanish mackerel (Scomberomorus niphonius) based on the otolith shape analysis in the Yellow Sea and Bohai Sea

Geographic variability in sagittal otoliths shape of the Japanese Spanish mackerel (Scomberomorus niphonius) was studied to test whether stock discrimination might be possible using this method. It was hypothesized that S. niphonius would show little difference in otolith shape, likely as a result of mixing between seas. A total of 164 sagittal otoliths were collected from commercial fishing vessels between April and June 2010 in three spawning grounds, that of Qingdao in the middle Yellow Sea, Lvsi in the southern Yellow Sea, Huanghua in the the Bohai Sea. To minimize the potential effect of the fish size among the three fishing grounds, a narrow size range (400–550 mm fork length) was selected. The discriminant function analysis conducted with standardized otolith shape indices and Fourier harmonics produced classification success rates ranging from 57.0 to 88.2% and worked well in the separation between the Bohai Sea group and the Yellow Sea group. No significant differences were detected among the three groups for the Lvsi spawning grounds. The otolith variables showed a distinct gradual variation tendency with the movement of fish schools from south to north. The results suggest that stock from the Bohai Sea could be managed as a separate entity from those found in the Yellow Sea.     

Otolith shape: a population marker for Atlantic herring Clupea harengus

Otolith shape variation of seven Atlantic herring Clupea harengus populations from Canada, the Faroe Islands, Iceland, Ireland, Norway and Scotland, U.K., covering a large area of the species' distribution, was studied in order to see if otolith shape can be used to discriminate between populations. The otolith shape was obtained using quantitative shape analysis, transformed with Wavelet and analysed with multivariate methods. Significant differences were detected among the seven populations, which could be traced to three morphological structures in the otoliths. The differentiation in otolith shape between populations was not only correlated with their spawning time, indicating a strong environmental effect, but could also be due to differing life‐history strategies. A model based on the shape differences discriminates with 94% accuracy between Icelandic summer spawners and Norwegian spring spawners, which are known to mix at feeding grounds. This study shows that otolith shape could become an accurate marker for C. harengus population discrimination.     

The use of otolith shape to determine stock structure of <Emphasis Type="Italic">Engraulis encrasicolus</Emphasis> along the Tunisian coast

Engraulis encrasicolus is of great economic importance in the Mediterranean. However, little is known about its stock structure. Otolith shape analysis has been successfully used for fish stock identification. In this study, the stock structure of anchovy caught off the open sea and the coastal area of the Gulf of Tunis, lagoon of Bizerte and Lake of Ichkeul were investigated using otolith shape. Otolith shape was determined by Fourier analysis and then compared among specimens sampled from different areas with forward stepwise canonical discriminant analysis. Significant differences in otolith shape between the open sea and inshore anchovy groups were detected. Otolith shape of anchovy collected in the Lake of Ichkeul was distinct from the other groups. This finding suggests a clear discreteness of the open sea and the continental groups. The data highlighted the potential for using otolith shape analysis for anchovy stock identification, as well as the role of oceanographic features in determining stock separation. These findings will have major implications for anchovy fisheries management in Tunisia. By using a precautionary approach and considering the three areas as separate stocks, fisheries management strategies should be adjusted to achieve optimum sustainable production from each stock and to avoid decreases in genetic variety.     

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.     

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.     

Otolith Microstructure of Larval Gymnocypris potanini Herzenstein from the Minjiang River in China

Synopsis Otolith microstructure of about 120 Gymnocypris potanini larvae from the Minjiang River in China was examined and analyzed. Larvae had multiple primordia in most lapilli and sagittae, while had only one primordium in a few specimens. There had only one nucleus in otoliths of the larvae, except for some few specimens with 2 nuclei. The transparence of many otoliths differed from center to edge, and part of them could be divided into inner low optically dense zone (LODZ) and outer optically dense zone (ODZ). Based on increment clarity, otoliths of this species could be classified into three types, which were otolith with subtile increments, otolith with almost identified increments, and otolith with fairly clear increments characterized by high contrast. The last two types of otolith accounted for 87.07% in lapilli and 94.46% in sagittae, respectively. Increment clarity of sagitta was higher than that of lapillus. Natural checks were identified in 32.50% lapilli and 48.33% sagittae. These checks primarily located in the first to sixth increment. According to the number of increments in otoliths, the age of this batch larvae was 14 – 22 days, birth date was on June 17 – 25, and average growth rate of body length was 0.8936 ± 0.08769 mm/d.     

Molecular and morphological diversity in species of Kronichthys (Teleostei,Loricariidae) from Atlantic coastal rivers of Brazil

The Neotropical catfish genus Kronichthys contains three species distributed along coastal rivers of southern and southeastern Brazil. Although phylogenetic hypotheses are available, the molecular and morphological diversity and species boundaries within the genus remain unexplored. In this study, the authors generated mitochondrial data for 90 specimens combined with morphometric and meristic data to investigate species diversity, species boundaries and putative morphological signatures in Kronichthys. Phylogenetic and species delimitation results clearly show the presence of four genetic lineages, three within Kronichthys heylandi along the coast from Rio de Janeiro to southern São Paulo and a single lineage encompassing both the nominal species Kronichthys lacerta and Kronichthys subteres from the Ribeira de Iguape basin to Santa Catarina in southern Brazil. Nonetheless, morphological data show overlapped ranges in morphometrics and a definition of only two morphotypes, with clear phenotypic differences in the teeth number: K. heylandi differs from K. subteres + K. lacerta by the higher number of premaxillary teeth (30–52 vs. 19–28) and higher number of dentary teeth (28–54 vs. 17–28). Headwater captures and connections of paleodrainages because of sea-level fluctuations represent the two major biogeographic processes promoting species diversification and lineage dispersal of Kronichthys in the Atlantic coastal range of Brazil.     

Zum Wachstum der Otolithen bei Jungheringen

Zusammenfassung Etwa 1000 Heringslarven und Jungheringe aus den Fängen der Jahre 1957 und 1958 wurden untersucht. Die Proben bestanden in erster Linie aus den Abkömmlingen der im Sommer/Herbst laichenden Bankheringe der mittleren und nördlichen Nordsee. Downslarven und -jungheringe spielten daneben nur eine geringe Rolle. Die Ausmessung des Otolithen und seines Zentralfeldes erfolgte auf Grund von Photographien.Die Ausformung des Otolithen wird anhand von Umrißzeichnungen beschrieben (Abb. 3).In den Abb. 4–6 sind die Allometrieverhältnisse Otolithenlänge : Totallänge, Kopflänge : Totallänge und Otolithenlänge : Kopflänge dargestellt. Die Allometrieverhältnisse von Kopf und Otolith zur Körperlänge ändern sich während des ersten Lebensjahres erheblich. Otolith und Kopf wachsen dagegen nach der Metamorphose annähernd isometrisch zueinander.Die Ausmessung des Zentralfeldes stößt infolge seiner oft unscharfen Begrenzung auf erhebliche Schwierigkeiten. Bei vielen Otolithen liegt zwischen dem eigentlichen Zentralfeld und der ersten opaken Zone eine semi-opake Übergangszone. Die Strukturen der Otolithen von Heringslarven werden kurz beschrieben.Nach Abschluß der Zentralfeldbildung scheint sich die Größe des Zentralfeldes — zumindest während der ersten beiden Lebensjahre — nicht zu verändern (Abb. 10). Längen-Rückenberechnungen an Zentralfeldern und direkte Beobachtungen ergaben, daß die Larven des Bankherings Ende März in der Deutschen Bucht mit der Bildung des ersten opaken Sommerringes beginnen.Der erste opake Ring des Heringsotolithen besteht wie bei anderen Fischen aus einer Aufeinanderfolge von Zonen unterschiedlicher Transparenz. Im Frühjahr werden sehr kräftige opake Strukturen angelegt, denen später transparentere folgen. Für manche Otolithen — besonders bei Jungheringen, die im August im Küstenwasser gefangen worden waren — war es nicht zu entscheiden, ob die diffuse hyaline Zone, die bereits im Sommer angelegt wird, als Sekundärring oder als erster Winterring bewertet werden muß.Weitere Untersuchungen über die Struktur der Otolithen von Heringen verschiedener Herkunft und über das Wachstum der Jungheringe der Deutschen Bucht sind in Vorbereitung.

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Summary This preliminary communication deals with the development of the herring otolith mainly during the first year of life. The material was collected in 1957 and 1958 within the German Bight. It mainly consists of the offspring of summer/autumn spawning Bank herring.The different developmental stages of otoliths of O-group herrings are shown in fig. 3. The typical outline of the herring otolith is reached at about 90 mms of total length. The otolith length (largest diameter) was determined from photographs. The allometry of the otolith growth to body size shows four main different phases: 1. The larval stage up to metamorphosis, 2. the growth during and immediately after metamorphosis, 3. a disturbance at about 80–90 mms of total length and 4. after that a growth similar to that of the larval stage (fig. 4). These conditions are about the same as in the relation of head to body length (fig. 5). Therefore the relation between head and otolith remains constant in the main. Only one significant break of the curve was found: Up to metamorphosis the otolith grows negatively allometric. Afterwards isometry is nearly established (fig. 6). The measurements on size of the otolith centre were hampered by the lack of a sharp contour of the centre in many otoliths. A transition zone lying between the centre and the opaque ring and the structure of larval otoliths is described (fig. 7–8).There is no difference between the size of otolith centres just after they have been formed and about 1 year later (fig. 10). Back calculations and direct observations showed, that the formation of the first opaque ring starts in larvae of about 28–30 mms at the end of March.The first opaque zone of the herring otolith consists of a succession of opaque and hyaline rings. In some otoliths it is not possible to come to a decision wether the broad zone of semi-opaque or hyaline elements following the first strong opaque ring has not to be reckoned as secondary ring meaning a part of the summer zone followed by a further opaque zone in late summer or rather as first winter ring.Further investigations are planned on otoliths of different origin and on the growth pattern of young herring in the German Bight.

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(Mt 10 Abbildungen und 2 Tabellen im Text)

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Ein Teil der zu diesen Untersuchungen verwendeten Ausrüstung wurde aus Mitteln der Deutschen Wissenschaftlichen Kommission für Meeresforschung angeschafft.     

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.     

Mn<Superscript>2+</Superscript> concentrations in coastal fish otoliths: understanding environmental and biological influences from EPR

The Mn²⁺ concentrations in the sagittae otoliths of 12 fish families (and 19 species) that co-occur in a coastal area of southeastern Brazil (~21°S) were quantified using electron paramagnetic resonance (EPR). Inferences were made about the relationship between fish habitat and trace element incorporation. Inferences were made on the relationship between trace element concentration and otolith shape. The differences in Mn²⁺ concentrations among the species suggest that habitat (and feeding habits) might drive the incorporation of this trace element into fish otoliths, with higher values in bottom-associated fish species than in surface-associated species. In surface-associated fish species, the correlation between trace element concentrations and otolith shape was stronger than in bottom-associated species. Thus, while the Mn bioavailability in a fish’s habitat, especially from feeding resources, is a local driving influence of trace element incorporation in sagittae otoliths, species-specific requirements also have an influence. Quantitative EPR is a non-destructive technique that is very useful when the available samples cannot be damaged, like with otolith collections.