New polymorphic microsatellite loci for population studies in the European anchovy, Engraulis encrasicolus (L.)

Eleven microsatellite loci were developed in the European anchovy, Engraulis encrasicolus, and tested in samples from two geographically distant populations (Atlantic and Mediterranean Sea). Number of alleles ranged from eight to 28 and observed heterozygosity from 0.440 to 0.920. There was no evidence of linkage disequilibrium, although two loci are indeed linked. All loci were in Hardy-Weinberg equilibrium, except for one locus in the Atlantic and two loci in the Mediterranean sample. These three loci plus two more showed evidence for null alleles.     

Field mortality estimates of anchovy larvae, Engraulis encrasicolus, in the western Mediterranean

Eggs and larvae of anchovy were obtained from plankton collections made along the Catalan coast (north-western Mediterranean) from 1983 to 1985 in order to determine spawning seasons and areas and other aspects of its life history. Previous studies have shown the existence of two main spawning areas, to the north and south of the Catalan coast, respectively. The northern one is characterized by a shorter spawning period related with the thermal cycle.
Mortality rates of larvae aged 4.9-15 days were calculated from the peak spawning month (June) of the three years, separately by spawning areas. Data on abundance by length classes were adjusted for escape and net avoidance and for duration of growth through a size class as derived from otolith readings. Mortality rates ranged from 0.17 to 0.58. Mortality was higher in 1983 than in 1984 and 1985, coinciding with a high production of anchovy eggs in that year. Mortality at the northern spawning area was lower than at the southern one.     

Modelling the presence of anchovy Engraulis encrasicolus in the Aegean Sea during early summer, based on satellite environmental data

Acoustic and satellite environmental data as well as bathymetry data were used to model the presence of anchovy, Engraulis encrasicolus during early summer in the northern Aegean Sea (Eastern Mediterranean). Generalized Additive Models (GAMs) were used for modelling and subsequently applied in a predictive mode to identify those areas in the Greek Seas and the entire Mediterranean basin that could support species’ presence. Model results were evaluated with the estimation of Receiver Operating Characteristic (ROC)-plots as well as qualitatively, based on (a) acoustic data from concurrent studies in certain areas of the northern Aegean Sea that were not included in the estimation of the GAM model and (b) historical acoustic data from the central Aegean and Ionian Seas. Mapping the estimated environmental conditions in the Mediterranean basin indicated areas that generally agree with the known distribution grounds of anchovy, such as the straits of Sicily and coastal waters of Tunisia, areas in the Tyrrhenian Sea, the Adriatic Sea, the Gulf of Lions and the Catalan Sea. Guest editor: V. D. Valavanis Essential Fish Habitat Mapping in the Mediterranean     

Mitochondrial DNA Sequence Variation Suggests the Lack of Genetic Heterogeneity in the Adriatic and Ionian Stocks of Sardina pilchardus

A genetic stock structure analysis of 11 sardine samples from the Adriatic Sea and Ionian neighboring area was carried out through sequence variation analysis of a 307-bp cytochrome b gene fragment in order to identify self-recruiting units in the Adriatic Sardina pilchardus stock. The overall lack of genetic subdivision among samples detected by analysis of molecular variance, pairwise Φ_st values, and the exact test of population differentiation indicates this sardine stock is part of a larger self-recruiting population whose boundaries are larger than the investigated area. This conclusion is in agreement with preliminary allozymic and mitochondrial DNA restriction fragment length polymorphism data, but contradicts the previous identification of 2 subpopulations of sardines in the Adriatic Sea argued on morphologic differences, which could be rather attributed to different hydrographic or ecologic conditions occurring in different areas of the Adriatic Sea. The reduced gene flow observed between Adriatic-Ionian and Spanish sardine geographic samples (P < 0.001) suggests that reproductively isolated populations of sardines may occur in the Mediterranean Sea.     

Lipid biomarkers and trophic linkages between phytoplankton, zooplankton and anchovy (Engraulis encrasicolus) larvae in the NW Mediterranean

Identification and quantification of the main fatty acids inphytoplankton, zooplankton and Engraulis encrasicolus larvaewere used to study the relationship among the different trophiclevels. Three northwestern Mediterranean sea areas were studiedas representative of potentially heterogeneous environments.HPLC pigment analysis was performed to assess the contributionof different phytoplankton groups to total chlorophyll biomass.In spite of the physical variability, a homogeneous patternin phytoplankton composition among areas was found, with thePrymnesiophyceae representing more than half of the phytoplanktonchlorophyll biomass. This homogeneous spatial pattern was alsoreflected in the phytoplankton fatty acid proportions. Thus,significant amount of 14:0, 18:1(n-9) and 18:4(n-3) could berelated to the dominance of the prymnesiophytes; the low proportionof fatty acid markers of diatoms and dinoflagellates agreedwith the low abundances of those phytoplankton groups. Zooplanktonfatty acids also showed homogeneous proportions in the studiedareas, with a preferential accumulation of 16:1(n-7) and 20:5(n-3),and 22:6(n-3). This observation may indicate a preferentialgrazing on diatoms and dinoflagellates. Anchovy larvae displayeddiffering fatty acids contributions related to size. Large larvaecontained a higher proportion of polyunsaturated fatty acid(PUFA). Small larvae showed a high percentage of 18:1(n-9) and18:4(n-3) fatty acids, considered Prymnesiophyceae biomarkers.These results suggest an additional food resource for the anchovythat is complementary to a zooplankton diet, but probably ofprotozoan origin (i.e. microzooplankton). Significant logarithmicrelationships were found between dry mass and 20:5(n-3) and22:6(n-3) fatty acid proportions in E. encrasicolus larvae.These PUFA are considered essential for the fish-larvae development.     

Diet composition and feeding of European anchovy Engraulis encrasicolus in Kiel Bight,western Baltic Sea

Diet composition of the expanding southern species European anchovy Engraulis encrasicolus in the western Baltic Sea was investigated. Results revealed an interesting case of bentho‐pelagic coupling with potential implications for local fish species through competition for food resources.     

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.     

Growth and length–weight relationship of the juvenile anchovy, Engraulis encrasicolus, in the nursery ground (Zrmanja River estuary–eastern Adriatic Sea)

Growth and length–weight relationships of 2564 juvenile specimens of anchovy, Engraulis encrasicolus (Linnaeus, 1758), were examined over a 2‐year period (August 1989 to April 1991) in the Zrmanja River estuary. Fish sizes ranged from 4.4 to 12.5 cm total length and 0.5 and 13.8 g, corresponding to age (in years) 0+ (98%) and 1+ (2%). Length–weight regression coefficient (b) varied between 2.95 and 3.42 and the regression constant (a) between 2.1 and 6.0 (×10^?3). The von Bertalanffy growth equation of juvenile anchovy was: l_t = 13.2 (1 ? e^?0.82(t+0.5)). Maturity was reached in June in males above 8.2 cm and females above 9.0 cm total length.     

Genetic analysis of the populations of Japanese anchovy Engraulis japonicus from the Yellow Sea and East China Sea based on mitochondrial cytochrome b sequence

Japanese anchovy Engraulis japonicus was not only a commercial species but also a key link in the marine food chain in the Yellow Sea and East China Sea. However, in recent years, E. japonicus experienced a deep recession caused by over fishing. Therefore, determination of population genetic structure is essential to underpin resource recovery and to aid delineating and monitoring populations for fishery management. In this study, sequence variation of a 402-bp fragment of mitochondrial cytochrome b (cyt b) gene from ten populations of E. japonicus in the Yellow Sea and East China Sea was determined, detecting 73 haplotypes. Haplotype diversities and nucleotide diversities for the ten populations ranged from 0.4706 ± 0.1177 (D) to 0.9935 ± 0.210 (WH), and from 0.0014 ± 0.0013 (D) to 0.0090 ± 0.0053 (WH), respectively. Analysis of molecular variance (AMOVA) showed no significant differences at all hierarchical levels, and almost all F_ST values were non-significant, indicating no significant population genetic structure in the Yellow Sea and East China Sea. Two indistinct clades with low support values were detected and the analysis of haplotype frequency distribution showed no significant geographic differences among populations. These results supported the null hypothesis that E. japonicus within the Yellow Sea and East China Sea constituted a panmictic mtDNA gene pool. Based on the present study, the existence of separate genetic stocks in the Yellow Sea and East China Sea were not detected. Strong dispersal capacity of planktonic larvae, and ocean current could be the reasons for genetic homogeneity in this species in the Yellow Sea and East China Sea.     

Interannual fluctuations in recruitment and growth of the sardine,Sardinops melanostictus,in the sea of Japan and adjacent waters

Recruitment and growth of the sardineSardinops melanostictus fluctuated markedly in the Sea of Japan and adjacent waters between 1978 and 1993. Stock size was calculated using Virtual Population Analysis and average body length in each age class was determined by the number of annual rings on the scales. There is an inverse correlation between average water temperature at a depth of 50 m in the coastal area of the mainland of Japan in winter (January to March) and recruitmentR defined as the number of individuals at 1 year old. There is also an inverse correlation between spawning stock sizeE and reproductive success in (R/E). A multiple regression model using spawning stock size and water temperature in winter as independent variables can explain 73% of variance in reproductive success. It suggests that both density-dependent and density-independent factors perform important roles determining reproductive success. There is an inverse correlation between body length and stock size and this suggests that there is a density-dependent effect on the growth of the sardine.     

A Second Look at Mitochondrial DNA Variability in European Anchovy (Engraulis encrasicolus): Assessing Models of Population Structure and the Black Sea Isolation Hypothesis

Genetic architectures of marine fishes are generally shallow because of the large potential for gene flow in the sea. European anchovy, however, are unusual among small pelagic fishes in showing large differences among sub-basins and in harbouring two mtDNA phylogroups (‘A’ & ‘B’), representing 1.1–1.85 million years of separation. Here the mtDNA RFLP dataset of Magoulas et al. [1996, Mol. Biol. Evol. 13: 178–190] is re-examined to assess population models accounting for this subdivided population structure and to evaluate the zoogeographical origins of the two major phylogroups. Haplotype and nucleotide diversities are highest in the Ionian Sea and lowest in the Aegean and Black seas. However, this gradient is absent when ‘A’ and ‘B’ haplotypes are examined separately. Neither the self-sustaining nor the basin population models adequately describe anchovy population behaviour. Tests for neutrality, mismatch and nested clade analyses are concordant in depicting recent expansions of both phylogroups. Unimodel mismatch distributions and haplotype coalescences dating to the last (Eemian) interglacial (‘B’) and the Weichselian pleniglacial period (‘A’) indicate separate colonizations of the Mediterranean Basin. Phylogroup ‘A’ is unlikely to have arisen through continuous long-term isolation in the Black Sea because of climate extremes from displaced subpolar weather systems during the ice ages. Ancestors of both groups appear to have colonized the Mediterranean from the Atlantic in the late Pleistocene. Hence, zoogeographic models of anchovy in the Mediterranean must also include the eastern (and possibly southern) Atlantic.     

Comparison of mesenteric and tissue fat content in relation to sexual cycle of the sardine, Sardina pilchardus (Walb., 1792), in the eastern Middle Adriatic fishery grounds (Croatia)

The study objective was to analyse Sardina pilchardus (Walb., 1792) for mesenteric fat, which is easier to evaluate than the muscle lipid, for comparisons thereof with the sexual cycle and condition of the examined specimens. Fat reserves, moisture and the sexual cycle of sardines were studied from monthly random samples of purse seine catches from March 2004 to February 2005. A total of 1209 specimens were collected (668 males; 541 females). Total length and mass ranged from 13.0 to 19.0 cm and from 16.7 to 51.5 g, respectively. Lipid analyses in muscle tissues were done using the Soxhlet method. An inverse relation between fat content and the sexual cycle was established. Greatest fat quantities were observed in August (72%) during the gonadal resting phase; lowest values were noted in winter, and in spring during the spawning period peak. Moisture content and sexual cycle were positively correlated ( r  = 0.7913). Total lipids in sardine tissues were higher in females than in males; however, the moisture content was higher in males than in females.     

Nutrient-dependent variation in growth and longevity of the red bandfish, Cepola macrophthalma, in the Aegean Sea

Statistically significant regressions are presented between the maximum observed age, maximum gross weight, mean total length, mean gross weight, and mean weighted back-calculated total length at ages 3 and 4 years for red bandfish, Cepola macrophthalma , and the total nitrogen in three Aegean gulfs. In contrast, the regressions of all the above mentioned biological variables on phosphates in the three gulfs were not significant, a fact consistent with the view that nitrogen, as opposed to phosphorus, is the limiting factor in eastern Mediterranean marine ecosystems. After discussing possible genetic discontinuities between the three populations, being inferred from the paleoceanography of the study area, it is suggested that the three populations form a single gene pool expressing phenotypic plasticity shaped by the contrasting environmental conditions experienced.     

Population Dynamics,Growth and Productivity of Abra ovata (Mollusca,Bivalvia) in the Evros Delta (North Aegean Sea)

Monthly samples of Abra ovata were collected during February 1983-January 1984 in the Evros Delta (N. Aegean Sea). Population density (mean annual value = 2407.5 ind · m⁻²) was characterized by seasonal variation. An analysis of the length frequency distributions shows that one annual recruitment of juveniles (> 2 mm) occurred in October-January; and also that, throughout the year, two age groups existed in the population. One growth ring was formed on the shells of the oldest age group during July-August. Mean growth in shell length can be described by Bertalanffy function. A positive correlation existed between shell length and decalcified dry weight. Secondary production in A. ovata, calculated by the instantaneous growth method, showed a mean biomass of 29.221 g dry weight m⁻²yr⁻¹, a productivity of 17.086 g dry weight m⁻²yr⁻¹ and an annual turnover ratio of 0.59.     

Sex ratio, spawning season and size at maturity of red bandfish in the western Aegean Sea

The red bandfish Cepola rubescens is characterized by a normal gonochoristic sexual organization. The overall male: female ratio did not differ significantly ( x ²=4.57, P <0.05) from the theoretical 1: 1. However, the male: female ratio differed with season and length class and these differences are most probably related to sexual differences in growth rate, natural mortality rate and energetic cost of reproduction. Red bandfish spawns over an extended period, from late spring to mid autumn, with larger females spawning earlier than smaller ones. Males reach maturity at a larger total length, 299 mm, and age, 2.6 years, than females, 219 mm and 1.9 years respectively, a fact most probably indicating the existence of social structure in red bandfish. Finally, red bandfish matures at a comparatively smaller length and age than other fish species.     

Age, growth and mortality of red bandfish, Cepola macrophthalma (L.), in the western Aegean Sea (Greece)

Age, growth and mortality of red bandfish, Cepola macrophrhalma (L.), from the western Aegean Sea were studied using the otoliths from 1113 fish and the length and weight of 3351 fish. Two regions were established (north and south of the Euripos Strait). The red bandfish grows allometrically (slope of length-weight regressions ≤2) and relatively rapidly until age 4 or 5 years. It is proposed that this rapid linear growth represents an adaptation which evolved to reduce the vulnerability of red bandfish to whole-fish swallowers. Back-calculated lengths-at-age showed significant differences between regions and between sexes. Males were larger than females after age 2 years. Maximum age of northern fish was also higher than southern fish (8 and 5 years respectively). Southern fish grow at a faster rate (K=0.379) but to a significantly smaller size (L_x=424.8mm t.l. , W_x=25.1 g) than northern fish (K=0.214, L_x=676.1 mm t.l. , W_x=90.9 g). Mortality rates (total and natural instantaneous mortalities) of southern fish were also higher than northern fish. We suggest that these variations represent responses to the different conditions of temperature and food availability prevailing in the two regions.     

Genetic architecture of differences in oviposition preference between ancestral and derived populations of the seed beetle Acanthoscelides obtectus

We investigated the additive, dominance and epistatic genetic effects underlying differentiation in oviposition preference between two populations of the seed beetle Acanthoscelides obtectus evolved in the laboratory for 102 generations on bean and chickpea seeds. We reared and tested females on each of two host legumes. The populations differed in mean oviposition preference; the preference for chickpea was stronger in population reared on the chickpea (C) than in population maintained on common bean (P). Observations in the parental populations indicated that females tend to prefer ovipositioning their eggs on the seeds they have already experienced. The patterns of the means in each of the parental populations and 12 types of hybrids (two F(1), two F(2) and eight backcrosses) indicated that population differences in oviposition preference from both rearing hosts could be explained by nonadditive genetic effects. Statistically detectable additive and dominance genetic effects were observed in the most parsimonious model only when females were reared on the chickpea. The most parsimonious models on both rearing hosts suggested a contribution of negative additive x additive epistasis to the divergence of oviposition preference between the P and C populations. This indicates a positive effect of epistasis on the performance of the second generations of hybrids.     

The presence of the potentially toxic genera Ostreopsis and Coolia (Dinophyceae) in the North Aegean Sea, Greece

The examination of macrophyte, water and sediment samples, collected at depths less than 1.5 m from 50 different sites along the North Aegean coasts, has revealed, for the first time in Greek coastal waters, the presence of two Ostreopsis species (O. ovata and O. cf. siamensis) and Coolia monotis in the majority of the sampling sites (94% and 100%, respectively). Other epiphytic dinoflagellates of the genera Prorocentrum and Amphidinium and diatoms were accompanying species in this epiphytic community. Morphometric features, plate formula and thecal ornamentation were used for species identification. O. ovata cells were smaller in dorsoventral (DV) diameter and width (W) (26.18–61.88 μm and 13.09–47.60 μm, respectively) in comparison with O. cf. siamensis (35.70–65.45 μm and 23.80–49.98 μm, respectively). In contrast, the anterioposterior (AP) diameter of O. cf. siamensis was smaller (14.28–26.18 μm) resulting in DV/AP ≈ 3, whereas the above ratio for O. ovata was less than 2 (AP ranging between 14.28–35.70 μm). Moreover, the theca of O. ovata cells was ornamented with scattered pores, which fluctuated in a wider range (0.07–0.32 μm) than those of O. cf. siamensis (0.23–0.29 μm). Coolia monotis cells were almost round with average DV diameter 26.88 μm, AP 25.66 μm and width 26.76 μm. Small and large cells were recorded in both field and culture populations of Ostreopsis spp. and C. monotis, while hyaline cysts were observed for O. ovata. The presence of O. ovata and O. cf. siamensis exhibited a clear seasonal pattern dominating (maximum abundance up to 4.05 × 10⁵ cells gr⁻¹ fwm) the period from midsummer to late autumn in years 2003 and 2004, while C. monotis was found also in winter and spring months.     

A direct approach to the measurement of genetic variation in fish populations: applications of the polymerase chain reaction to studies of Atlantic cod, Gadus morhua L.

We used the polymerase chain reaction (PCR) and direct DNA sequencing to study genetic variation within and among populations of Atlantic cod, Gadus morhua , in the western North Atlantic. In a 307 bp region of the mitochondrial cytochrome b gene, 24 variable nucleotide positions define 24 genotypes, which differ by from one to six nucleotide substitutions. Greenland cod ( G. ogac ) differs from the most similar G. morhua genotype by an additional 12 nucleotide substitutions. Silent transitions dominate both intra- and interspecific comparisons, however four nucleotide substitutions within morhua result in amino acid replacements. Direct sequencing of DNA reveals substantially more of the genetic variation that exists within and between species than do previous indirect methods based on restriction fragment length polymorphisms, and thus has far greater potential to quantify such differences as may exist among fish stocks. Preliminary experiments also indicate that automation of DNA sequencing provides an efficient, rapid, and accurate means for detection of genetic variation in natural populations offish.