Genetic differentiation of Mediterranean horse mackerel (Trachurus mediterraneus) populations as revealed by mtDNA PCR-RFLP analysis

The genetic population structure of Mediterranean horse mackerel, Trachurus mediterraneus , from seven locations throughout the Black, Marmara, Aegean and eastern Mediterranean seas was investigated using restriction fragment length polymorphism (RFLP) analysis of the mtDNA 16S rDNA region. An approximately 2000-bp segment was screened in 280 individuals using six restriction enzymes, resulting in 10 composite haplotypes. The most common haplotype was present in 56.42% individuals; the next most frequent haplotype was present in 22.85% individuals. Average haplotype diversity within samples was moderate (0.38), and nucleotide diversity was low (0.00435). Mean nucleotide divergence for the seven sampling sites was 0.0028. Nucleotide divergence among samples was moderate, with the highest value detected between the Aegean Sea (Izmir) and the eastern Black Sea (Trabzon) populations (0.007055), and the lowest (−0.000043) between the Marmara Sea (Adalar) and the western Black Sea (Sile) populations. In Monte Carlo pairwise comparisons of haplotype frequencies, the Sinop from the middle Black Sea, Trabzon from the eastern Black Sea, and Iskenderun Bay from the north-eastern Mediterranean Sea exhibited highly significant (P   <   0.001) geographical differentiation from each other and from all other populations. Mantel's test indicated that the nucleotide divergence among populations of T. mediterraneus was not significantly associated with their geographical isolation ( r  = −0.2963; P   >   0.05). Consequently, the mtDNA 16S rDNA region provided evidence for the existence of three distinct T. mediterraneus populations (Sinop, Trabzon and Iskenderun Bay) in the Black and north-eastern Mediterranean seas.     

Genetic and morphological analyses of tub gurnard Chelidonichthys lucerna populations in Turkish marine waters

Genetic and morphological structure of tub gurnard Chelidonichthys lucerna populations in Turkish marine waters were investigated with mtDNA sequencing of 16S rRNA and morphological characters. C. lucerna samples were collected from the Black Sea, Marmara, Aegean and northeastern Mediterranean coasts of Turkey. The lowest genetic diversity was found in the northeastern Mediterranean (Iskenderun Bay) population, while the highest was in the Marmara population with overall average value of genetic diversity within populations. A total of 14 haplotypes was found, and the highest haplotype diversity was in the Black Sea whereas the lowest was in the northeastern Mediterranean population (Iskenderun Bay). The Black Sea and Iskenderun Bay populations showed the least genetic divergence (0.001081), while the highest was between the Marmara Sea and northeastern Mediterranean (Antalya Bay) populations (0.002067). Pairwise comparisons of genetic distance revealed statistically significant differences (P < 0.05) between the Marmara and both the Aegean and northeastern Mediterranean (Antalya Bay) samples. Neighbour joining tree analyses clustered the northeastern Mediterranean populations (Antalya Bay and Iskenderun Bay) as genetically more interrelated populations, whereas the Aegean Sea population was clustered as most isolated one. Discriminant function analysis of morphological characters showed that only the Black Sea population is differentiated from the other populations.     

Interspecific variations of otolith chemistry in estuarine fish nurseries

Otolith chemical composition differed between juveniles of five fish species ( Solea solea , Solea senegalensis , Platichthys flesus , Diplodus vulgaris and Dicentrarchus labrax ) in nursery areas of six estuaries along the Portuguese coast (Douro, Ria de Aveiro, Mondego, Tejo, Sado and Mira). Spatially consistent patterns in the concentration of some elements were responsible for differences between species. Discrimination of estuaries using data from all species simultaneously averaged 44·7% of correctly classified cases, whilst discrimination of species averaged 76·2%. Moreover, species-related patterns in otolith fingerprints were highlighted when comparing species for each estuarine nursery area, with intra estuarine species discrimination averages ranging from 86·2 to 100·0%. Similarities in the otolith elemental fingerprints were larger between species with close phylogeny and ecology, particularly between flatfish and perciform species. In addition to the differences in physiological regulation of species, specific microhabitat use in a common environment was suggested as a relevant factor for the differentiation of otolith chemistry among species occurring in the same locations. Despite positive results in specific estuaries, variation in otolith composition limited the use of species as proxies to classify others to their system of origin.     

Microsatellite DNA reveals genetically different populations of Atlantic bonito Sarda sarda in the Mediterranean Basin

Microsatellites were used to investigate population genetic structure of Atlantic bonito Sarda sarda from the Black Sea, Marmara Sea, Aegean Sea, north-eastern Mediterranean Sea and Adriatic Sea. Overall average observed heterozygosity was high (0.93). Average observed heterozygosity per locus ranged from 0.79 to 0.98. Pairwise F_ST estimates for all loci between populations ranged from 0 to 0.07626, and significant F_ST values (P < 0.001) were detected between populations; the Blacks Sea and Marmara Sea samples were not significantly different from each other, but significant different from the other samples, and Aegean Sea and north-eastern Mediterranean Sea samples were also not significantly different from each other, but significantly different from all other samples. The Adriatic Sea sample was significant different from all other samples. The Mantel test revealed a significant (P < 0.001, r = 0.68) isolation-by-distance for these 11 populations. Neighbour-joining analysis clustered the Black Sea and Marmara Sea samples together while collections from Aegean Sea and north-eastern Mediterranean Sea were clustered close to each other and far from the others. On the other hand, the Adriatic Sea collection presented very distinctive relationship from the others.     

Isolation by distance and selection effects on genetic structure of sardines Sardina pilchardus Walbaum

Allozyme data were used to analyse the genetic structure of Sardina pilchardus populations. Fifty samples from 15 locations between the North Sea and Mauritania, including samples from the Azores, Madeira and the Mediterranean Sea, were surveyed. A weak but significant structure was found between all samples ( F _ST= 0.057, P < 0·001). This structure results from a change in the most common allele of SOD* between the North African and the Azores populations separated by the greatest distance. This locus seemed to be under selective pressure according to the test of neutrality, and the variations in allele frequencies may be explained due to isolation by distance (IBD) of coastal populations (from Mauritania to the North Sea) ( r = 0·86, P < 0·001). When SOD* was removed from the analyses, IBD was not observed in coastal populations ( r = 0·236, P > 0·05) but was observed over the whole range ( r = 0·321, P = 0·05). The genetic structure of S. pilchardus is driven by both IBD and selective processes.     

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.     

Genetic structuring of European anchovy (Engraulis encrasicolus) populations through mitochondrial DNA sequences

Mitochondrial DNA sequence variation in 655?bpfragments of the cytochrome oxidase c subunit I gene, known as the DNA barcode, of European anchovy (Engraulis encrasicolus) was evaluated by analyzing 1529 individuals representing 16 populations from the Black Sea, through the Marmara Sea and the Aegean Sea to the Mediterranean Sea. A total of 19 (2.9%) variable sites were found among individuals, and these defined 10 genetically diverged populations with an overall mean distance of 1.2%. The highest nucleotide divergence was found between samples of eastern Mediterranean and northern Aegean (2.2%). Evolutionary history analysis among 16 populations clustered the Mediterranean Sea clades in one main branch and the other clades in another branch. Diverging pattern of the European anchovy populations correlated with geographic dispersion supports the genetic structuring through the Black Sea-Marmara Sea-Aegean Sea-Mediterranean Sea quad.     

First documentation of the otoliths of the species of Gouania (Teleostei: Gobiesocidae) in the Mediterranean Sea

Otolith morphology is a widely accepted tool for species identification in teleost fish, but whether this holds true for very small species remains to be explored. Here, the saccular otoliths of the cryptobenthic Mediterranean clingfish Gouania (Gobiesocidae) are described for the first time. The new data, although preliminary, indicate that otolith morphology and morphometry support the recognition of the recently differentiated five species of Gouania in the Mediterranean Sea. Furthermore, otoliths of phylogenetically closely related Gouania species resemble each other more than do those of the more distantly related species.     

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.     

Temperature effects on the incorporation of strontium in otolith of Japanese eel Anguilla japonica

The effects of temperature on somatic and otolith growth and the incorporation of strontium in otolith of the Japanese eel, were studied in laboratory-reared and field-caught eels. The somatic and otolith growth rates of the eel increased significantly with temperature and were estimated as approximately 0·096 mm t.l, (P<0·01) and 0·36 μm in otolith diameter per degree-day (0·01相似文献   

Mitochondrial DNA reveals a mosaic pattern of phylogeographical structure in Atlantic and Mediterranean populations of anchovy (Engraulis encrasicolus)

This study extends the geographic coverage of a previous study of mitochondrial DNA restriction fragment length polymorphism in European anchovy. Both studies together include 24 samples representing 17 localities extending from the Black Sea, through the Mediterranean Sea to the eastern Atlantic as far south as Dakar, Senegal. Eighty-eight haplotypes define two clades (A and B) separated by 3.2% sequence divergence. Clade A has a star-like genealogy indicative of a recent population expansion. Clade B has a more complex genealogy, consisting of several haplotypes at intermediate frequencies. The distributions of these clades consist of a mosaic with abrupt changes between some areas and gradients between other areas. Clade A predominates the Black and Aegean seas, but is present throughout the Mediterranean. Unexpectedly, new data show that clade A is also at a high frequency in the Atlantic, from Portugal to at least Senegal. Overall, the level of genetic differentiation among populations is high (F(ST)=0.148, p<0.0001), with the greatest differences between basins. AMOVA reveals four main geographical groups: Atlantic, central Mediterranean, Aegean Sea, and Black Sea. Mismatch distribution clearly indicates historical bottleneck and population expansion for clade A, while for clade B such evidence is equivocal. This difference may reflect a range expansion for both clades, but with higher gene flow (Nm values) between demes for clade A. Both contemporary and historical processes are important in shaping the complex genetic population structure of European anchovy.     

Comparison of otolith growth and morphology with somatic growth and age in young-of-the-year bluefin tuna

Otolith morphological characteristics were studied using image analysis techniques and the relationships between otolith growth and somatic growth and age, as estimated from counting daily otolith increments, were examined in young-of-the-year (YOY) bluefin tuna Thunnus thynnus ranging in fork length ( L _F) from 8·5 to 55·5 cm. Whole otolith length, width, area and perimeter, and three shape indexes, circularity, E value and rectangularity, were extracted for each pair of sagittae. Since no statistical significant differences between left and right otolith morphometrics were found, only one otolith from each fish was used for correlations. Statistically significant relationships were observed between otoliths measurements and fish somatic growth when a linear regression was applied after logarithmic transformation of all variables tested. Among the variables, otolith length was the one that showed the highest correlation with L _F, followed by otolith area and perimeter, whereas otolith rectangularity exhibited the lowest correlation. Statistically significant relationships were also observed between the otolith variables tested and the age of the fish, which ranged from 20 to 129 days. The ages estimated using otolith mass were very close to those assessed using daily increment counts (bias ranged from 1 to 24 days). Therefore, otolith mass could represent a valuable criterion for age estimation in YOY bluefin tuna that is objective, economic and easy to perform compared to daily increment counting method.     

Genetic differentiation among populations of the shortfinned eel Anguilla australis from East Australia and New Zealand

Variability at seven microsatellite loci was used to survey the genetic population structure of the shortfinned eel Anguilla australis . Samples were collected from six estuaries along the east coast of Australia and from three estuaries around New Zealand. Hierarchical analysis of molecular variance of the five loci with good fit to Hardy–Weinberg genotypic proportions detected highly significant differences among samples ( F _ST= 0·016, P < 0·001). The fixation index between countries ( F _CT= 0·012, P < 0·001) was more than double the index among samples within countries ( F _SC= 0·005, P < 0·05). An unweighted pair-group method with arithmetic mean (UPGMA) tree also supported the separation of Australian and New Zealand populations, as did assignment tests, which correctly assigned 80 and 84% of the individuals to Australia and New Zealand, respectively. Isolation-by-distance appeared among samples overall ( r = 0·807, P < 0·001), but not among samples within countries ( r = 0·027, P > 0·05 in Australia; r = 0·762, P > 0·05 in New Zealand). These findings indicate that populations of A. australis in East Australia and in New Zealand may be reproductively isolated from one another. Genetic differentiation among populations of A. australis was two- to 10-fold higher than that among populations of other temperate eels in the North Atlantic Ocean, suggesting that two group of A. australis may reflect sub-species. Anguilla australis in the two countries have different genetic structures and thus require separate management. Genetic isolation between Australian and New Zealand populations indicates that juveniles recruit independently into these two regions from geographically or temporally isolated spawning areas.     

An eco-morphological explanation of individual variability in the shape of the fish otolith: comparison of the otolith of Hoplostethus atlanticus with other species by depth

Variation in otolith shape (otolith polymorphism) in Hoplostethus atlanticus, Hoplostethus mediterraneus, Paratrichichthys trailli, Pagrus major and Trachurus murphyi , quantified using principal components analysis based on Fourier transform decompositions of outlines of otolith shape, was particularly high among the samples of H. atlanticus from the North Atlantic, New Zealand, Australia and Namibia. The scatter was uniform, however, and did not show any significant differences among regions. The implication drawn from the high variability in otolith shape of H. atlanticus was that otolith shape polymorphism was maintained by some form of balancing selection across many small local environments which may result in k-selection with consequent poor response by H. atlanticus to maximum sustainable yield harvesting strategies. The variation in otolith shape defined by the otolith morphospace of the five species that were measured, showed a decreasing trend in scatter (i.e. decreasing complexity of shape) proceeding from the species with the deepest habitat ( H. atlanticus ) to the most shallow ( T. murphyi ).     

Discrepancies between otoliths of larvae and juveniles of the American eel: is something fishy happening at metamorphosis?

The use of otoliths to interpret early life history in fishes depends upon the assumptions that otoliths record past events accurately and consistently and that records of events in otoliths are continuous. Both the number of growth microincrements ( I ), and the radii ( R ,μm) of otoliths of American eel Anguilla rostrata , leptocephali increased linearly and highly significantly with leptocephalus body length ( L , mm), as expected on the above assumptions ( I , =2·29 L , − 5·75 and R , =1·05 L , + 12·02, r ²,=0·938 and 0·931, n , =20). In contrast, the number of increments and the radii of the leptocephalus growth zones of otoliths of glass-phase American eels were not related to body length, and they were lower than predicted by the relationships developed for leptocephali. Thus, otoliths of American eels apparently violate one or both assumptions. Possibly, the margin of the otolith is resorbed during metamorphosis from leptocephalus to glass eel, perhaps as part of calcium metabolism as skeletal elements are being formed.     

Latitudinal Gradient in Otolith Shape among Local Populations of Atlantic Herring (Clupea harengus L.) in Norway

Otolith shape analysis of Atlantic herring (Clupea harengus) in Norwegian waters shows significant differentiation among fjords and a latitudinal gradient along the coast where neighbouring populations are more similar to each other than to those sampled at larger distances. The otolith shape was obtained using quantitative shape analysis, the outlines were transformed with Wavelet and analysed with multivariate methods. The observed morphological differences are likely to reflect environmental differences but indicate low dispersal among the local herring populations. Otolith shape variation suggests also limited exchange between the local populations and their oceanic counterparts, which could be due to differences in spawning behaviour. Herring from the most northerly location (69°N) in Balsfjord, which is genetically more similar to Pacific herring (Clupea pallasii), differed in otolith shape from all the other populations. Our results suggest that the semi-enclosed systems, where the local populations live and breed, are efficient barriers for dispersal. Otolith shape can thus serve as a marker to identify the origin of herring along the coast of Norway.     

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.     

The relationship between somatic growth and otolith dimensions of Mediterranean horse mackerel (Trachurus mediterraneus) from the Black Sea

In this study the relationships between somatic growth and otolith dimensions of Mediterranean horse mackerel (Trachurus mediterraneus) were investigated from samples obtained in the southeastern Black Sea area. It was concluded that there is a significant relationship between fish size and otolith dimensions. Therefore, age of Mediterranean horse mackerel can be estimated from otolith length and weight with sufficient certainty. This research was the first study of these relationships for T. mediterraneus from the Black Sea.     

Age and growth of the dusky grouper Epinephelus marginatus (Lowe 1834) in an exploited population of the western Mediterranean Sea

The age and growth of the dusky grouper, Epinephelus marginatus , in the Balearic Islands (western Mediterranean) were studied by otolith analysis from a sample of 358 specimens ranging in total length ( L _T) from 6·6 to 105·6 cm. The specimens came from commercial artisanal and recreational spear fisheries between 1999 and 2003. Otoliths grew asymmetrically throughout the range of L _T studied, showing a clear pattern of alternating translucent and opaque bands. Marginal increment analysis of specimens up to 8 years-old indicated that a single opaque band was formed each year during spring and summer. Whole otoliths allowed ageing specimens up to 10 years old, but above that age whole otoliths yielded lower age estimates than sectioned otoliths. The maximum estimated age was 61 years, which significantly extends the estimated life span of the species from a maximum of 36 years in a previous study. The von Bertalanffy growth parameters were estimated as L _∞= 95·5 cm L _T, K = 0·087 and T _O=−1·12. The study revealed differences in mean L _T at age and age structure between the shallow- and deep-water samples which may be attributed to different fishing pressure and environmental conditions.     

The taxonomic status of the Black Sea and Marmara Sea populations of the Broadnosed Pipefish Syngnathus cf. argentatus Pallas (Teleostei: Syngnathidae) based on morphological and molecular characters

As the Black Sea and Marmara Sea population of the Broadnosed Pipefish Syngnathus cf. argentatus show some morphological differences from the Mediterranean Sea populations, some authors regard it as an endemic species Syngnathus argentatus Pallas, 1814, while others consider it as a synonym of S. typhle Linnaeus, 1758. The aim of this study is to compare the populations of the Black Sea and Sea of Marmara with the Aegean Sea population, using a combination of morphological and molecular characters, in order to clarify their taxonomic status. Sampling was carried out at three stations in the Black Sea, two in the Sea of Marmara and three in the Aegean Sea, and a total of 24 morphometric and 6 meristic characters were examined. Metric data were analysed by Principal Component Analysis (PCA) and phylogenetic relationships between the populations were analysed using both cytochrome oxidase subunit 1 (COI) gene and cytochrome b (cyt-b) gene sequences. Although constant differences were observed in snout depths between the Black Sea/Marmara Sea and the Aegean Sea populations, other morphological features and genetic analysis did not enable these populations to be differentiated. These findings indicate that S. argentatus is a synonym of S. typhle.