Spatiotemporal variation in the population structure of the European hake in the NW Mediterranean

The bathymetric distribution of Merluccius merluccius was studied as a function of length, age and maturity of specimens caught by commercial trawl and longline in Different seasons. Males matured first at 28·8 cm (3 years old), and females at 38·0 cm (3·5 years old). Reproductive activity was noted practically throughout the year with its most pronounced spawning peak in the autumn. Most fish <37–40 cm were males and >46 cm were females. Specimens occurred between 50 and 750 m depth, although density was low at >400 m. Adults were found at all depth strata studied. Recruits and juveniles were limited to inshore waters <400 m, most were found between 100 and 200 m. Spring and summer were the preferred seasons for recruitment; although for both seasons there was some interannual variation. Adult distribution also varied, according to the season. Young adults were spread over the entire depth range, with the biggest ones concentrated at the edge of the shelf (150–350 m), especially in autumn and winter. The main spawning peak coincided with this concentration of adults suggesting that spawning occurred in autumn/winter at the edge of the shelf.     

First record of microplastics ingestion by European hake MERLUCCIUS MERLUCCIUS from the Tyrrhenian Sicilian coast (Central Mediterranean Sea)

A sample of 67 European hake Merluccius merluccius were examined to highlight the ingestion of microplastics in the Tyrrhenian Sea. In all samples, 31 black fibres were found in the stomach contents corresponding to 46.3% of the specimens. The data presented here could be important for the implementation of the EU Marine Strategy Framework Directive in Mediterranean waters.     

Temporal and spatial genetic variation in spawning grounds of European hake (Merluccius merluccius) in the Bay of Biscay

Polymorphism at five microsatellite loci were screened to determine the genetic variability and the temporal stability of population structure in natural populations of European hake (Merluccius merluccius, L.) within the Bay of Biscay. In addition, the control region (900 bp) and two protein coding genes (ATPase, subunits 6 and 8, 842 bp and a partial sequence of the ND1, 800 bp) of the mitochondrial DNA (mtDNA) were sequenced from geographically distant populations from the extremes of the species range. One hundred individuals from either side of a supposed stock boundary within the bay were collected in autumn 1997. This sampling strategy was repeated during hake spawning seasons in late spring of 1998 and 1999. Low levels of population subdivision were found between putative populations within years. Similarly, low levels of differentiation were found between autumn 1997 northern samples and spring 1998 southern samples which were collected 7 months later on spawning grounds. These results are discussed in relation to ecological, behavioural and oceanographic information. Sampling effects, which may influence these results, are also discussed. Theta (theta) estimates were significantly different from zero in every other pairwise comparison between geographical areas (north and south of the Bay of Biscay) and between years within the same area (P<0.05). Hierarchical analysis of molecular variance (AMOVA) does not confirm the temporal persistence of population structure. These results are discussed in relation to variance in reproductive success, and temporal spawning patterns, which may exist within the bay. mtDNA variability was very low between geographically distant samples from Norway and the Mediterranean Sea with only 10 variable sites found in a total of 2542 bp of mtDNA, these differences being exclusively in the D-loop.     

Macrogeographical population differentiation in oceanic environments: a case study of European hake (Merluccius merluccius), a commercially important fish

Polymorphism at six microsatellite loci was used to study genetic variability and population structure in six geographically distant natural populations of European hake (Merluccius merluccius L.). Four hundred and eighty-three individuals were sampled from Trondheimsfjord in Norway, the Celtic Sea, the southern Bay of Biscay, Faro off Portugal, the Mediterranean Sea north of the coast of Tunisia and the Adriatic Sea. Population subdivision was found between Mediterranean and Atlantic samples, theta = 0.029 (P < 0. 001). No substructuring was found between samples within the Mediterranean Sea, theta = 0.003 and RST = 0.007 (P > 0.05). The Atlantic population structure appears to be more complex than previously suggested by the placement of stock boundaries by the International Council for the Exploration of the Seas (ICES). Analyses based on various models of microsatellite evolution all suggest that differentiation exists between Bay of Biscay and Portugese samples, theta = 0.013 (P < 0.001), RST = 0.036 (P < 0. 001) which are currently managed as one stock. By contrast, fixation indices indicated no differentiation between southern Bay of Biscay samples and Celtic Sea samples, theta = 0.003 (P = 0.02), phiST = 0. 007 (P = 0.10) which are managed as separate stocks. These results suggest that if the observed trends are stable through time, current management policy of European hake may need revision.     

Parasites of Cynoscion guatucupa along South American Atlantic coasts: evidence for stock discrimination

Two stocks of striped weakfish Cynoscion guatucupa in the south‐west Atlantic Ocean, were identified using parasites as biological tags. A total of 297 fish caught in Brazil, Uruguay and Argentina, contained 29 species of metazoan parasites. Univariate analysis on parasite populations, as well as multivariate discriminant analysis, calculated for juvenile and adult fish separately, allowed the identification of the two stocks, one from Argentina and Uruguay and the other from Brazil, to be made. Southern samples were characterized by higher prevalences and abundance of larval endohelminths, whereas in the northern stock, gastrointestinal and ectoparasitic species were common. Discriminant analyses on parasite infracommunities of Brazilian fish showed notable differences between juvenile and adult hosts; no such trend was observed in fish from Argentina and Uruguay, despite differences between juveniles and adults at the population level. Different oceanographic conditions and their influence on the distribution of parasites as well as of other hosts involved in their life cycles could be key factors for the differences observed among stocks of C. guatucupa .     

Molecular identification of Anisakis simplex sensu stricto and Anisakis pegreffii (Nematoda: Anisakidae) from fish and cetacean in Japanese waters

Parasites morphologically consistent with Anisakis simplex sensu lato collected from the coast of Japan and Western North Pacific Ocean were examined by PCR-RFLP of the ITS region (ITS1, 5.8 subunit rRNA gene and ITS2) and mtDNA cox1. The RFLP patterns of rDNA generated by HinfI and HhaI showed that 100% of the larvae collected from Hokkaido and 94% of adults collected from Western North Pacific Ocean were identified as A. simplex sensu stricto. On the other hand, 97% of the larvae collected from Fukuoka prefecture were identified as A. pegreffii. A hybrid genotype was found in adults in Western North Pacific Ocean and larva in Fukuoka prefecture. These findings revealed that A. simplexs. str. is primarily distributed in the North Pacific Ocean and A. pegreffii is primarily distributed in the southern Sea of Japan. RFLP analysis of mtDNA cox1 showed different patterns between A. simplex s. str. and A. pegreffii after digestion with HinfI. This polymorphism obtained by RFLP analysis of mtDNA cox1 proved the usefulness as new genetic markers to distinguish two sibling species.     

Ontogenetic shift and feeding habits of the European hake (Merluccius merluccius L., 1758) in Central and Southern Tyrrhenian Sea (Western Mediterranean Sea): A comparison between past and present data

The present paper aims to investigate the ecological role of Merluccius merluccius, Linnaeus, 1758, in southern and central Tyrrhenian Sea (GSA 10, Resolution GFCM/33/2009/2 General Fisheries Commission for the Mediterranean), analyzing ontogenetic diet shifts, geographical variations on prey composition, and feeding habits. A total of 734 hake specimens ranging in size between 6 cm and 73 cm (Total Length, TL) were collected in 2018. In order to evaluate ontogenetic shifts in prey composition, samples were divided into five size classes and for each class the quantitative feeding indices have been calculated. The statistical analysis, based on index of relative importance percentage (%IRI), resulted in three trophic groups. The most abundant prey found in the immature hake specimens (size class I) were the Euphausiids, Stylocheiron longicorne and Mysidacea, while for samples with a total length over 10.5 cm were crustaceans and fish. Engraulis encrasicolus was the most abundant fish prey identified, followed by Boops boops and Myctophids. The high presence of Euphausiids, Mysids, Myctophidae, and Sternoptychidae in classes I, II, II, and IV (6–23 cm) showed the relevant role of mesopelagic fauna in hake diets, with an essential organic matter and energy flow from the mesopelagic to the epipelagic environment. Additionally, decapod crustaceans were found in the stomach contents of hakes belonging to class V (with size over 36 cm TL), which is notable considering that our study area includes an important decapod crustacean fishing area.     

Morphological and molecular characterization of Anisakis larvae (Nematoda: Anisakidae) in Beryx splendens from Japanese waters

The third-stage (L3) larvae of Anisakis, which are the etiological agents of human anisakiasis, have been categorized morphologically into Anisakis Type I larvae and Anisakis Type II larvae. Genetic analysis has allowed easy identification of these larvae: Anisakis Type I larvae include the species Anisakis simplex sensu stricto, Anisakis pegreffii, Anisakis simplex C, Anisakis typica, Anisakis ziphidarum, and Anisakis nascettii, whereas Anisakis Type II larvae include the species Anisakis physeteris, Anisakis brevispiculata, and Anisakis paggiae. Since human consumption of raw fish and squid is common in Japan, we investigated Anisakis L3 larvae in 44 specimens of Beryx splendens from Japanese waters. A total of 730 Anisakis L3 larvae collected from B. splendens were divided morphologically into 4 types: Type I, Type II, and 2 other types that were similar to Anisakis Type III and Type IV described by Shiraki (1974). Anisakis Type II, Type III, and Type IV larvae all had a short ventriculus, but their tails were morphologically different. In addition, data from genetic analysis indicated that Anisakis Type II, Type III, and Type IV larvae could be identified as A. physeteris, A. brevispiculata, and A. paggiae, respectively. Therefore, A. physeteris, A. brevispiculata, and A. paggiae can be readily differentiated not only by genetic analysis but also by morphological characteristics of L3 larvae.     

Electrophoretic studies on the Anisakis simplex complex (Ascaridida:Anisakidae) from the Mediterranean and North-East Atlantic

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and 1986. Electrophoretic studies on the Anisakis simplex complex (Ascaridida: Anisakidae) from the Mediterranean and North-East Atlantic. International Journal for Parasitology 16: 633–640. The genetic variation of the sibling species Anisakis simplex A and A. simplex B was investigated by electrophoretic analysis of 22 gene-enzyme systems. The two species are reproductively isolated and no gene flow takes place between them. Three loci, Sod, Adk-2 and Lap-1, show distinct alleles in A. simplex A and A. simplex B, allowing their reliable identification both at the larval and adult stages. A fourth locus, Got, appears to be diagnostic at the 95% level. The value of Nei's genetic distance found between A. simplex A and A. simplex B is 0.28. Parameters of genetic variability (He, P, A) are given for both species. The geographic distribution of A. simplex A and A. simplex B appears to be mainly Mediterranean for the former, and mainly North Atlantic for the latter. Several paratenic hosts (fish and squid) and one cetacean definitive host are identified for each of the two species. The names A. pegreffii and A. simplex are tentatively proposed for A. simplex A and A. simplex B respectively.     

Parasites of Argentine anchovy in the south-west Atlantic: latitudinal patterns and their use for discrimination of host populations

Four stocks of Argentine anchovy Engraulis anchoita in the south-west Atlantic, south of 34°S were identified by the use of parasites as biological tags. A total of 671 fish were examined for parasites and 19 parasite species were found in 664 Argentine anchovies. The parasite populations and both infra- and component communities showed marked latitudinal gradients. Cluster analyses allowed the identification of four groups, one composed of autumn samples (35–37°S) and three of spring ones. Among the latter, a Patagonian group (43–46°S) was clearly separated from a Bonaerense group, which was divided into two groups: North Bonaerense (34–40°S) and South Bonaerense (40–43°S). Autumn samples were characterized by low values of prevalence for most parasite species. Among spring samples Pseudanthocotyloides heterocotyle , Lecithochirium microstomum , Parahemiurus merus , Scolex polymorphus and Corynosoma australe were characteristic of the northern region, whereas Cardiocephaloides sp., Anisakis simplex , Contracaecum sp. and Hysterothylacium aduncum were more prevalent in southern waters. Latitudinal environmental gradients and their influence on the distribution of both zooplankton and definitive hosts, could be determinants of the clines observed.     

Identification of fish species by an egg identification key: an alternative to the incubation of eggs and rearing of larvae for species recognition

Comparison of the results of two methods for species recognition, an egg identification key and rearing of larvae, showed only small qualitative differences in the identification of fish species present in a stretch of the River Elbe, Germany. Both the identification key and rearing methods complement each other as methods of identification, and the selection of which method to be used should depend primarily on the aim of the investigation.     

Metazoan parasites of Sebastes capensis from two localities in northern Chile as tools for stock identification

Examination of 290 red rockfish Sebastes capensis from two fishing grounds in northern Chile (Coquimbo 71°30' W; 30°0' S and Antofagasta 70°40' W; 23°30' S) revealed 8770 metazoan parasites, belonging to 18 taxa: Udonella caligorum (Udonellidae); Interniloculus chilensis , Paramicrocotyle sp. and Neobenedenia melleni (Monogenea); larval Gnathia sp., Cirolana sp. and Rocinela sp. (Isopoda); Helicometrina nimia , Diphtherostomum sp., Lecithochirium sp. and Pseudopecoelus sp. (Digenea); Ascarophis sebastodis , Anisakis sp. and larval Hysterothylacium sp. (Nematoda); Caligus cheilodactylus , Lepeophtheirus chilensis and Trifur tortuosus (Copepoda); larval Corynosoma australe (Acanthocephala). Ten species were found only in Coquimbo and three were found only in Antofagasta, species common to both localities were Pseudopecoelus sp., C. australe , Anisakis sp. and A. sebastodis . Evidence, based on qualitative differences between the parasite faunas of the red rockfish, suggests the existence of two discrete stocks.     

Seasonal stability and spatial variability of parasites in Brazilian sandperch Pinguipes brasilianus from the Northern Argentine Sea: evidence for stock discrimination

The use of parasites as biological tags allowed the identification of two stocks of Brazilian sandperch Pinguipes brasilianus , in the Bonaerense region of the Argentine Sea. A total of 156 adult specimens of P. brasilianus were examined for parasites. Temporal variability in parasite burdens was assessed from fish caught seasonally in coastal waters of two zones, south Bonaerense (during autumn and winter) and north Bonaerense (during summer). Additional data from a previous study, comprising P. brasilianus caught during spring in these two zones, as well as in two populations from north Patagonian gulfs were used for comparative analyses of spatial variability. A total of 14 193 metazoan parasites belonging to 19 species were found. Comparisons of seasonal variability in pooled samples and within each locality showed that locality effects exceeded seasonal ones, suggesting the possible existence of two discrete stocks in the Bonaerense region. These findings were strongly supported by discriminant analyses and comparisons of prevalence and abundance between zones, after pooling seasonal samples within each zone. Further evidence of the discreteness of both stocks was assessed by inclusion of samples from Patagonian gulfs in the discriminant analysis, confirming that their differences were at a inter-population level. The parasite species that contributed most to the separation of the samples were generally those identified as biological markers in previous studies. Differing oceanographic conditions are discussed as potential causes of inter-population variation of parasite burdens.     

Genetic signatures in an invasive parasite of Anguilla anguilla correlate with differential stock management

In this article, it is shown that available genetic tools for the omnipresent parasite Anguillicoloides crassus in European eels Anguilla anguilla are sensitive to different immigration rates into local A. anguilla stocks for two separated river systems. Relying on four highly polymorphic microsatellite markers, it was inferred that under natural recruitment, nematode samples meet Hardy–Weinberg expectations for a single panmictic population, while genetic signals show signs for a strong Wahlund effect most likely due to very recent population mixing under frequent restocking of young A. anguilla. This was indicated by a low but significant F_ST value among within‐host populations (infrapopulations) along with high inbreeding indices F_IS consistent over all loci. The latter signal is shown to stem from high levels of admixture and the presence of first‐generation migrants, and alternative explanations such as marker‐ and sex‐specific biases in the nematode populations could be dismissed. Moreover, the slightly increased degree of relatedness within infrapopulations in the stocked river system cannot explain the excessive inbreeding values found and are most likely a direct consequence of recent influx of already infected fish harbouring parasites with different genetic signatures. Applying a simulation approach using known variables from the nematode's invasion history, only the artificial introduction of a Wahlund effect leads to a close match between simulated and real data, which is a strong argument for using the parasite as a biological tag for detecting and characterizing fish translocation.     

Genetic composition of Atlantic and Mediterranean recruits of European eel Anguilla anguilla based on EST-linked microsatellite loci

Anguilla anguilla glass eels arriving at two Mediterranean and two Atlantic sites were tested for differences in genetic composition between regions using a total of 23 microsatellite loci developed from an expressed sequence tag (EST) library. Hierarchical analysis of molecular variance indicated a non-significant difference between regions (Mediterranean v . Atlantic), which contrasted with the significant differences observed between samples within regions. The existence of a single spawning site for all A. anguilla individuals and extensive migration loop with great opportunity for mixing of individuals might explain the homogeneity in genetic composition found between regions. The observation of a (small-scale) pattern of genetic patchiness among intra-annual samples (arrival waves) within geographic regions does not conflict with the lack of (large-scale) geographic sub-structuring found between the Mediterranean and Atlantic regions, but most likely is a consequence of the strong dependence of A. anguilla on oceanic conditions in the Sargasso Sea that might result in a limited parental contribution to each spawning event. The comparison of Atlantic and Mediterranean A. anguilla glass eel recruits based on EST-linked microsatellite loci provides evidence supporting the hypothesis of panmixia A. anguilla across Europe.     

Use of parasite and genetic markers in delineating populations of winter flounder from the central and south‐west Scotian Shelf and north‐east Gulf of Maine

Discriminant function analyses of infection parameters of parasitic helminths revealed that abundances of seven helminth species contributed significantly to the delineation of four host populations of winter flounder Pleuronectes americanus from the central and south‐west Scotian Shelf and the north‐east Gulf of Maine (NAFO subdivision 4WX‐5Z). These were adult digeneans, Derogenes varicus , Genolinea laticauda , Steganoderma formosum and Steringophorus furciger , metacercariae of the digenean, Stephanostomum baccatum , and larval nematodes, Anisakis simplex and Hysterothylacium aduncum . The correct classification rate was 84% overall, with Georges Bank (5Z) and Sable Island Bank (4W) winter flounder being the most accurately classified samples at 98 and 88%, respectively. Winter flounder from south‐west Nova Scotia (4X), an inshore sample from St Marys Bay and offshore fish from Browns Bank, had the lowest rates of correct classification (76 and 71%, respectively) due, primarily, to cross‐misclassification between the two samples. Winter pairwise comparisons of four microsatellite markers identified significant genetic differences between all populations sampled with the Georges Bank population being the most genetically distinct overall, and St Marys Bay and Browns Bank fish being the least dissimilar.     

Distribution of Anisakis larvae, identified by genetic markers, and their use for stock characterization of demersal and pelagic fish from European waters: an update

In the present paper, recent results obtained on the use of different distributions observed in larval species of Anisakis, genetically identified by means of allozyme markers, for stock characterization of demersal (Merluccius merluccius), small (Trachurus trachurus) and large pelagic (Xiphias gladius) finfish species in European waters, are reviewed and discussed. Several species of Anisakis were identified in the three fish hosts: A. simplex (s.s.), A. physeteris, A. typica, A. ziphidarum, A. pegreffii, A. brevispiculata and A. paggiae. Canonical discriminant analysis performed on all the samples of the three fish species collected in areas comprising their geographical range, according to the different species of Anisakis identified, showed distinct fish populations in European waters. In all the three fish hosts, the pattern of distribution of Anisakis larvae allowed discrimination of Mediterranean stocks from Atlantic stocks. In the case of swordfish, the possible existence of a southern Atlantic stock separated from a northern one is also suggested. Congruence and discordance with the population genetic data inferred from allozyme markers on the same samples of the three fish species are also discussed.     

Comparison of the feeding apparatus and diet of European sardines Sardina pilchardus of Atlantic and Mediterranean waters: ecological implications

In this study, the feeding apparatus (gill rakers, GR) and the diet composition of European sardine Sardina pilchardus populations living in two contrasting environments were compared: the upwelling area off western Iberia and the comparatively less productive region of the north‐western Mediterranean Sea. The importance of local adaptations in the trophic ecology of this species was estimated. Sardina pilchardus from the Atlantic Iberian coast and from the north‐western Mediterranean Sea have clear differences in the feeding apparatus and diet compositions. Those from the Atlantic Iberian coast have significantly more GRs than S. pilchardus of the same size range in the Mediterranean Sea. While S. pilchardus from the Mediterranean Sea mostly depend on prey ranging between 750–1500 and 3000–4000 µm, corresponding mostly to cladocerans, decapods and copepods, those from the Atlantic depend on smaller prey (50–500 and 1000–1500 µm) that include phytoplankton and copepods, particularly during summer months, and S. pilchardus eggs during the winter. The marked difference between the trophic ecology of S. pilchardus in the two areas studied appears to have originated from different dietary strategies that the two populations have adopted in contrasting feeding environments. These differences are shown to profoundly affect the size and quality of prey consumed, and the effect of cannibalism on the populations.