Redundancy analysis, genome-wide association studies and the pigmentation of brown trout (Salmo trutta L.).

The association of molecular variants with phenotypic variation is a main issue in biology, often tackled with genome-wide association studies (GWAS). GWAS are challenging, with increasing, but still limited, use in evolutionary biology. We used redundancy analysis (RDA) as a complimentary ordination approach to single- and multitrait GWAS to explore the molecular basis of pigmentation variation in brown trout (Salmo trutta) belonging to wild populations impacted by hatchery fish. Based on 75,684 single nucleotide polymorphic (SNP) markers, RDA, single- and multitrait GWAS allowed the extraction of 337 independent colour patterning loci (CPLs) associated with trout pigmentation traits, such as the number of red and black spots on flanks. Collectively, these CPLs (i) mapped onto 35 out of 40 brown trout linkage groups indicating a polygenic genomic architecture of pigmentation, (ii) were found to be associated with 218 candidate genes, including 197 genes formerly mentioned in the literature associated to skin pigmentation, skin patterning, differentiation or structure notably in a close relative, the rainbow trout (Onchorhynchus mykiss), and (iii) related to functions relevant to pigmentation variation (e.g., calcium- and ion-binding, cell adhesion). Annotated CPLs include genes with well-known pigmentation effects (e.g., PMEL, SLC45A2, SOX10), but also markers associated with genes formerly found expressed in rainbow or brown trout skins. RDA was also shown to be useful to investigate management issues, especially the dynamics of trout pigmentation submitted to several generations of hatchery introgression.

A genomic perspective on an old question: Salmo trouts or Salmo trutta (Teleostei: Salmonidae)?

There are particular challenges in defining the taxonomic status of recently radiated groups due to the low level of phylogenetic signal. Members of the Salmo trutta species-complex, which mostly evolved during and following the Pleistocene, show high morphological and ecological diversity that, along with their very wide geographic distribution, have led to morphological description of 47 extant nominal species. However, many of these species have not been supported by previous phylogenetic studies, which could be partly due to lack of significant genetic differences among them, the limited resolution offered by molecular methods previously used, as well as the often local scale of these studies. The development of next-generation sequencing (NGS) and related analytical tools have enhanced our ability to address such challenging questions. In this study, Genotyping-by-Sequencing (GBS) of 15,169 filtered SNPs and mitochondrial DNA (mtDNA) D-loop sequences were combined to assess the phylogenetic relationships among 166 brown trouts representing 21 described species and three undescribed groups collected from 84 localities throughout their natural distribution in Europe, west Asia, and North Africa. The data were analysed using different clustering algorithms (admixture analysis and discriminant analysis of principal components-DAPC), a Bayes Factor Delimitation (BFD) test, species tree reconstruction, gene flow tests (three- and four-population tests), and Rogue taxa identification tests. Genomic contributions of the Atlantic lineage brown trout were found in all major sea basins excluding the North African and Aral Sea basins, suggesting introgressive hybridization of native brown trouts driven by stocking using strains of the Atlantic lineage. After removing the phylogenetic noise caused by the Atlantic brown trout, admixture clusters and DAPC clustering based on GBS data, respectively, resolved 11 and 13 clusters among the previously described brown trout species, which were also supported by BFD test results. Our results suggest that natural hybridization between different brown trout lineages has probably played an important role in the origin of several of the putative species, including S. marmoratus, S. carpio, S. farioides, S. pellegrini, S. caspius (in the Kura River drainage) and Salmo sp. in the Danube River basin. Overall, our results support a multi-species taxonomy for brown trouts. They also resolve some species in the Adriatic-Mediterranean and Black Sea drainages as members of very closely related genomic clusters that may need taxonomic revision. However, any final conclusions pertaining to the taxonomy of the brown trout complex should be based on an integrative approach combining genomic, morphological, and ecological data. To avoid challenges in taxonomy and conservation of species complexes like brown trouts, it is suggested to describe species based on genomic clusters of populations instead of describing species based only on morphologically differentiated single type populations.

Development of a large SNPs resource and a low-density SNP array for brown trout (Salmo trutta) population genetics.

BACKGROUND: The brown trout (Salmo trutta) is an economically and ecologically important species for which population genetic monitoring is frequently performed. The most commonly used genetic markers for this species are microsatellites and mitochondrial markers that lack replicability among laboratories, and a large genome coverage. An alternative that may be particularly efficient and universal is the development of small to large panels of Single Nucleotide Polymorphism markers (SNPs). Here, we used Restriction site Associated DNA sequences (RADs) markers to identify a set of 12,204 informative SNPs positioned on the brown trout linkage map and suitable for population genetics studies. Then, we used this novel resource to develop a cost-effective array of 192 SNPs (96 × 2) evenly spread on this map. This array was tested for genotyping success in five independent rivers occupied by two main brown trout evolutionary lineages (Atlantic -AT- and Mediterranean -ME-) on a total of 1862 individuals. Moreover, inference of admixture rate with domestic strains and population differentiation were assessed for a small river system (the Taurion River, 190 individuals) and results were compared to a panel of 13 microsatellites. RESULTS: A high genotyping success was observed for all rivers (< 1% of non-genotyped loci per individual), although some initially used SNP failed to be amplified, probably because of mutations in primers, and were replaced. These SNPs permitted to identify patterns of isolation-by-distance for some rivers. Finally, we found that microsatellite and SNP markers yielded very similar patterns for population differentiation and admixture assessments, with SNPs having better ability to detect introgression and differentiation. CONCLUSIONS: The novel resources provided here opens new perspectives for universality and genome-wide studies in brown trout populations.

Genomic consequences of a recent three-way admixture in supplemented wild brown trout populations revealed by local ancestry tracts.

Understanding the evolutionary consequences of human-mediated introductions of domesticated strains into the wild and their subsequent admixture with natural populations is of major concern in conservation biology. However, the genomic impacts of stocking from distinct sources (locally derived vs. divergent) on the genetic integrity of wild populations remain poorly understood. We designed an approach based on estimating local ancestry along individual chromosomes to provide a detailed picture of genomic admixture in supplemented populations. We used this approach to document admixture consequences in the brown trout Salmo trutta, for which decades of stocking practices have profoundly impacted the genetic make-up of wild populations. In southern France, small local Mediterranean populations have been subject to successive introductions of domestic strains derived from the Atlantic and Mediterranean lineages. To address the impact of stocking, we evaluate the extent of admixture from both domestic strains within populations, using 75,684 mapped SNPs obtained from double-digested restriction site-associated DNA sequencing. Then, the chromosomal ancestry profiles of admixed individuals reveal a wider diversity of hybrid and introgressed genotypes than estimated using classical methods for inferring ancestry and hybrid pedigrees. In addition, the length distribution of introgressed tracts retained different timings of introgression between the two domestic strains. We finally reveal opposite consequences of admixture on the level of polymorphism of the recipient populations between domestic strains. Our study illustrates the potential of using the information contained in the genomic mosaic of ancestry tracts in combination with classical methods based on allele frequencies for analysing multiple-way admixture with population genomic data.

Salmo macrostigma (Teleostei, Salmonidae): Nothing more than a brown trout (S. trutta) lineage?

We examined specimens of the macrostigma trout Salmo macrostigma, which refers to big black spots on the flanks, to assess whether it is an example of taxonomic inflation within the brown trout Salmo trutta complex. Using new specimens, publicly available data and a mitogenomic protocol to amplify the control and cytochrome b regions of the mitochondrial genome from degraded museum samples, including one syntype specimen, the present study shows that the macrostigma trout is not a valid species. Our results suggest the occurrence of a distinct evolutionary lineage of S. trutta in North Africa and Sicily. The name of the North African lineage is proposed for this lineage, which was found to be sister to the Atlantic lineage of brown trout, S. trutta.

Phylogeography of the European sturgeon (Acipenser sturio): A critically endangered species.

The European sturgeon (Acipenser sturio) was once a common species throughout Europe, but the sole remaining natural population presently inhabits the Gironde Estuary in France (Atlantic coast). The species was classified as 'Critically Endangered' in 1996, and the Gironde population is now on the verge of extinction. In this setting, and for the first time, we present the past phylogeographical features of this species throughout Europe along with an assessment of its former genetic diversity. This study was based on a molecular analysis (mtDNA CR sequencing) of 10 living specimens from the Gironde Estuary, 55 museum specimens that had been caught along 19th and 20th centuries, and 59 archaeological remains dating back to 260-5000years BP, from which mitochondrial DNA was extracted and amplified. Although discontinuous, the produced data provided a realistic image of the former structure of A. sturio in Europe. Reconstruction of the phylogenetic trees and haplotypes network led to the identification of several clades. The mitochondrial genetic diversity of this species was found to be much greater at the core (Iberian Peninsula, Mediterranean and Adriatic regions) than along the margins (Atlantic-Northern Europe, Black Sea) of its range. A series of hypotheses on the dates and causes of changes in the species' major structures are put forward on the basis of these data. Finally, competition with A. oxyrinchus, a sibling species whose presence in Northern Europe was recently reconsidered, is presented as a major factor in the evolution of this species.

Evolution of Polyploidy and Functional Diploidization in Sturgeons: Microsatellite Analysis in 10 Sturgeon Species.

Sturgeons (family Acipenseridae) are one of the most endangered groups of animals. Two hundred million years of evolution and multiple ploidy levels make this group a unique subject for studying the evolution of polyploidy in animals. As most sturgeon species have gone through significant functional diploidization, 2 scales of ploidy levels can be distinguished: the "evolutionary scale," which indicates the maximum ploidy level achieved and the "recent scale," which indicates the current functional ploidy level. This study analyzes published and new microsatellites to check the ploidy level and to determine the degrees of functional diploidization in 10 sturgeon species from Europe and Asia. We screened 50 primer pairs newly developed for Acipenser gueldenstaedtii and 40 primer pairs previously developed in other studies for other sturgeon species. The maximal number of alleles per individual of a given species was assessed at 20 microsatellite loci, which showed consistent amplification in most of the 10 analyzed species. Taken together, our data on the percentage of disomic loci in different species suggest that functional diploidization is an ongoing process in sturgeons. We observed lower levels of diploidization in tetraploid species from the Atlantic clade than in the species from the Pacific clade, which can be explained by the more recent genome duplication in tetraploid species from the Atlantic clade. Based on the recent findings and results of this study, we propose that the evolution of sturgeons has been affected by at least 3 different polyploidization events.

Genetic diversity and conservation of the Prespa trout in the Balkans.

The Balkans are known to have a high level of biodiversity and endemism. No less than 15 taxa have been recorded in salmonids of the Salmo genus. Among them, the Prespa trout is found in only four river systems flowing into Lake Macro Prespa, three in the Former Yugoslav Republic of Macedonia and one in Greece. This is the first comprehensive survey of all streams located within the Macro Prespa Basin, encompassing the whole taxon range. A large genetic sample of 536 Prespa trout was collected mainly between 2005 and 2007. The sampling included 59 individuals from the Golema river system, 93 from the Kranska, 260 from the Brajcinska, 119 from the Agios Germanos, and five individuals from the lake itself. These specimens were analyzed with six microsatellite markers and by sequencing the mitochondrial control region. Nuclear data were examined through multidimensional analysis and assignment tests. Five clusters were detected by assignment: Golema, Kranska, Brajcinska upstream, Rzanska Brajcinska tributary and Brajcinska downstream. Most of these river systems thus hosted differentiated Prespa trout populations (with past gene flows likely dating before the construction of dams), except Agios Germanos, which was found to be composed of 5% to 32% of each cluster. Among the five trout individuals from the lake, four originated from Kranska River and one was admixed. Supported parsimonious hypotheses are proposed to explain these specificities. Conservation of this endemic taxon should take these results into account. No translocation should be performed between different tributaries of the lake and preservation of the Brajcinska populations should address the upstream-downstream differentiation described.

Evolutionary history of a widespread Indo-Pacific goby: the role of Pleistocene sea-level changes on demographic contraction/expansion dynamics.

Compared to endemics, widespread species are of particular interest to retrace recent evolutionary history. These species have a large population size which provides a clearer genetic signature of past events. Moreover, their wide geographic range increases the potential occurrence of evolutionary events (expansion, divergence, etc.). Here, we used several coalescent-based methods to disentangle the evolutionary history of a widespread amphidromous goby (Sicyopterus lagocephalus), in the light of sea-level variations during the Pleistocene. Using 75 samples recovered from three biogeographic regions (Western Indian Ocean, Melanesia and Polynesia), we analysed a portion of the cytochromeb gene and confirmed three major haplogroups, each specific to a region. Furthermore, we found that: (1) the Melanesian haplogroup was the oldest while the two peripheral regions hosted daughter haplogroups; (2) two centrifugal colonisation events occurred from Melanesia to the periphery, each synchronised with periods of strong paleo-ENSO episodes; (3) the demographic contraction-expansion events were linked to Pleistocene sea-level changes; (4) Melanesia and Polynesia acted as efficient refuges during the Last Glacial Maximum. These results highlight the importance of studying widespread species to better understand the role of climate changes and paleo-oceanography on the evolution of biodiversity.

Site heteroplasmy in the mitochondrial cytochrome b gene of the sterlet sturgeon Acipenser ruthenus.

Sturgeons are fish species with a complex biology. They are also characterized by complex aspects including polyploidization and easiness of hybridization. As with most of the Ponto-Caspian sturgeons, the populations of Acipenser ruthenus from the Danube have declined drastically during the last decades. This is the first report on mitochondrial point heteroplasmy in the cytochrome b gene of this species. The 1141 bp sequence of the cytb gene in wild sterlet sturgeon individuals from the Lower Danube was determined, and site heteroplasmy evidenced in three of the 30 specimens collected. Two nucleotide sequences were identified in these heteroplasmic individuals. The majority of the heteroplasmic sites are synonymous and do not modify the sequence of amino acids in cytochrome B protein. To date, several cases of point heteroplasmy have been reported in animals, mostly due to paternal leakage of mtDNA. The presence of specific point heteroplasmic sites might be interesting for a possible correlation with genetically distinct groups in the Danube River.

Phylogeographic structure and demographic patterns of brown trout in North-West Africa.

The objectives of the study were to determine the phylogeographic structure of brown trout (Salmo trutta) in Morocco, elucidate their colonization patterns in North-West Africa and identify the mtDNA lineages involved in this process. We also aimed to resolve whether certain brown trout entities are also genetically distinct. Sixty-two brown trout from eleven locations across the Mediterranean and the Atlantic drainages in Morocco were surveyed using sequence analysis of the mtDNA control region and nuclear gene LDH, and by genotyping twelve microsatellite loci. Our study confirms that in Morocco both the Atlantic and Mediterranean basins are populated by Atlantic mtDNA lineage brown trout only, demonstrating that the Atlantic lineage (especially its southern clade) invaded initially not only the western part of the Mediterranean basin in Morocco but also expanded deep into the central area. Atlantic haplotypes identified here sort into three distinct groups suggesting Morocco was colonized in at least three successive waves (1.2, 0.4 and 0.2-0.1 MY ago). This notion becomes more pronounced with the finding of a distinct haplotype in the Dades river system, whose origin appears to coalesce with the nascent stage of the basal mtDNA evolutionary lineages of brown trout. According to our results, Salmo akairos, Salmo pellegrini and "green trout" from Lake Isli do not exhibited any character states that distinctively separate them from the other brown trout populations studied. Therefore, their status as distinct species was not confirmed.

Nuclear markers of Danube Sturgeons hybridization.

Acipenseriformes are composed of 25 sturgeon species and two paddlefish species distributed exclusively in the northern hemisphere. The Danube River and the Black Sea were originally inhabited by six sturgeon species but two are extinct and only four are still reproducing currently in the Lower Danube: Huso huso, Acipenser stellatus, A. gueldenstaedtii and A. ruthenus. Sturgeon species hybridize more easily than other fish and the determination of pure species or hybrid status is important for conservation and for breeding in fish farms. This survey demonstrated that morphological determination of this status is not reliable and a molecular tool, based on eight microsatellites genotypes is proposed. This method, based on three successive statistical analyses including Factorial Correspondence Analysis (FCA), Structure assignation and NewHybrids status determination, showed a high efficiency in discriminating pure species specimens from F1, F2 and two kinds of backcross individuals involving three of the four reproducing Lower Danube sturgeon species.

Evidence of unidirectional gene flow in a fragmented population of Salmo trutta L.

Selection, genetic drift, and gene flow affect genetic variation within populations and genetic differences among populations. Both drift and selection tend to decrease variation within populations and increase differences among populations, whereas gene flow increases variation within populations but leads to populations being related. In brown trout (Salmo trutta L.), the most important factor in population fragmentation is disrupted river-segment connectivity. The main goal of the study was to use genetic analysis to estimate the level of gene flow among resident and migratory brown trout in potential hybridization areas located downstream of impassable barriers in one river basin in the southern Baltic Sea region. First, spawning redds were counted in the upper river basin downstream of impassable barriers. Next, samples were collected from juveniles in spawning areas located downstream of barriers and from adults downstream and upstream of barriers. Subsequently, genetic analysis was performed using a panel of 13 microsatellite loci and the Salmo trutta 5 K SNP microarray. The genetic differentiation estimated between the resident form sampled upstream of the barriers and the anadromous specimens downstream of the barriers was high and significant. Analysis revealed that gene flow occurred between the two forms in the hybridization zone investigated and that isolated resident specimens shared spawning grounds with sea trout downstream of the barriers. The brown trout population from the river system investigated was slightly, internally diversified in the area accessible to migration. Simultaneously, the isolated part of the population was very different from that in the rest of the basin. The spawning areas of the anadromous form located downstream of the barriers were in a hybridization zone and gene flow was confirmed to be unidirectional. Although they constituted a small percentage, the genotypes typical upstream of the barriers were admixed downstream of them. The lack of genotypes noted upstream of the barriers among adult anadromous individuals might indicate that migrants of upstream origin and hybrids preferred residency.

Phylogenetic relationships of hexaploid large-sized barbs (genus Labeobarbus, Cyprinidae) based on mtDNA data.

The phylogenetic relationships among species of the Labeobarbus genus (Teleostei, Cyprinidae) which comprises large body-sized hexaploid taxa were inferred using complete cytochrome b mitochondrial gene sequences. Molecular data suggest two main evolutionary groups which roughly correspond to a Northern (Middle East and Northwest Africa) and a sub-Saharan lineage. The splitting of the African hexaploids from their Asian ancestors and their subsequent diversification on the African continent occurred in the Late Miocene, a period in which other cyprinins also invaded Africa and radiated in the Mediterranean region. Finally, systematic implications of these results to the taxonomic validity of genera or subgenera such as Varicorhinus, Kosswigobarbus, Carasobarbus and Capoeta are further discussed.

Nuclear and mitochondrial DNA sequence data reveal the evolutionary history of Barbus (Cyprinidae) in the ancient lake systems of the Balkans.

Freshwater fauna of ancient lakes frequently contain endemic taxa thought to have originated during the long existence of these lakes, yet uncertainties remain as to whether they represent distinct genetic lineages with respect to more widespread relatives and to the relative roles of isolation and dispersal in their evolution. Phylogenetic analyses of sequence variation at nuclear and mitochondrial genes were used to examine these issues for the freshwater fish genus Barbus in two European ancient lake systems on the Balkan Peninsula. The nuclear and mitochondrial data yielded concordant phylogeographic patterns though incomplete sorting of nuclear haplotypes between some mitochondrial clades was detected. The distributions of two currently recognized species investigated here do not match the distributions of evolutionary lineages revealed by phylogenetic analyses. The Prespa barbel, Barbus prespensis, is not endemic to the lakes Prespa as previously thought but is instead found to be widespread in the south-eastern Adriatic Sea basin, with a distribution largely corresponding to the basin of the now extinct Lake Maliq historically connected with Lake Prespa. On the other hand, a cryptic phylogenetic subdivision in a widespread species, B. rebeli, was discovered to be more distant from B. rebeli than from other Barbus species and to be endemic to the system of connected lakes Ohrid and Shkodra. The division coincides with the hydrogeographical boundary delimiting distributions of other freshwater fishes, and we suggest that this newly discovered evolutionary lineage represents a distinct species. These findings support the emerging pattern that endemic taxa have evolved not through isolation of individual lakes, but in systems of currently and historically interconnected lakes and their wider basins.

Unsuspected intraspecific variability in the toxin production, growth and morphology of the dinoflagellate Alexandrium pacificum R.W. Litaker (Group IV) blooming in a South Western Mediterranean marine ecosystem, Annaba Bay (Algeria).

Physiological plasticity gives HABs species the ability to respond to variations in the surrounding environment. The aim of this study was to examine morphological and physiological variability in Alexandrium pacificum R.W. Litaker (Group IV) (former Alexandrium catenella) blooming in Annaba bay, Algeria. Monoclonal cultures of up to 30 strains of this neurotoxic dinoflagellate were established by the germination of single resting cysts from the surface sediment of this southern Mediterranean marine ecosystem. Ribotyping confirmed formally for the first time that A. pacificum is developing in Eastern Algerian waters. Toxin analyses of A. pacificum strains revealed substantial intraspecific variability in both the profile and toxin amount. However, the toxin profile of most strains is characterized by the dominance of GTX6 (up to 96 mol %) which is the less toxic paralytic molecule. The toxin concentrations in the isolated strains varied widely between 3.8 and 30.82 fmol cell-1. We observed an important variation in the growth rate of the studied A. pacificum strains with values ranging from 0.05 to 0.33 d-1. The lag time of the studied strains varied widely and ranged from 4 to 20 days. The intraspecific diversity could be a response to the selection pressure which may be exerted by different environmental conditions over time and which can be genetically and in turn physiologically expressed. This study highlights, for the first time, that the sediment of a limited area holds an important diversity of A. pacificum cysts which give when germinate populations with noticeable physiological plasticity. Consequently, this diversified natural populations allow an exceptional adaptation to specific environmental conditions to outcompete local microalgae and to establish HABs which could explain why this dinoflagellate is successful and expanding worldwide.

Unexpected genetic diversity among and within populations of the toxic dinoflagellate Alexandrium catenella as revealed by nuclear microsatellite markers.

Since 1998, blooms of Alexandrium catenella associated with paralytic shellfish poisoning have been repeatedly reported for Thau Lagoon (French Mediterranean coast). Based on data obtained for rRNA gene markers, it has been suggested that the strains involved could be closely related to the Japanese temperate Asian ribotype of the temperate Asian clade. In order to gain more insight into the origin of these organisms, we carried out a genetic analysis of 61 Mediterranean and 23 Japanese strains using both ribosomal and microsatellite markers. Whereas the phylogeny based on ribosomal markers tended to confirm the previous findings, the analysis of microsatellite sequences revealed an unexpected distinction between the French and Japanese populations. This analysis also highlighted great intraspecific diversity that was not detected with the classical rRNA gene markers. The Japanese strains are divided into two differentiated A. catenella lineages: the Sea of Japan lineage and the east coast lineage, which includes populations from the Inland Sea and the Pacific Ocean. A. catenella strains isolated from Thau Lagoon belong to another lineage. These findings indicate that microsatellite markers are probably better suited to investigations of the population genetics of this species that is distributed worldwide. Finally, application of the population genetics concepts available for macroorganisms could support new paradigms for speciation and migration in phytoplankton assemblages.

Genetic architecture of trout from Albania as revealed by mtDNA control region variation.

To determine the genetic architecture of trout in Albania, 87 individuals were collected from 19 riverine and lacustrine sites in Albania, FYROM and Greece. All individuals were analyzed for sequence variation in the mtDNA control region. Among fourteen haplotypes detected, four previously unpublished haplotypes, bearing a close relationship to haplotypes of the Adriatic and marmoratus lineages of Salmo trutta, were revealed. Ten previously described haplotypes, characteristic of S. ohridanus, S. letnica and the Adriatic and Mediterranean lineages of S. trutta, were also detected. Haplotypes detected in this study were placed in a well supported branch of S. ohridanus, and a cluster of Mediterranean-Adriatic-marmoratus haplotypes, which were further delimited into three subdivisions of Mediterranean, marmoratus, and a previously non-described formation of four Adriatic haplotypes (Balkan cluster). Haplotypes of the Balkan cluster and the other Adriatic haplotypes, do not represent a contiguous haplotype lineage and appear not to be closely related, indicating independent arrivals into the Adriatic drainage and suggesting successive colonization events. Despite the presence of marmoratus haplotypes in Albania, no marbled phenotype was found, confirming previously reported findings that there is no association between this phenotype and marmoratus haplotypes.

Parentage analysis with few contributing breeders: validation and improvement.

Validation of parental allocation using PAPA software (Duchesne P, Godbout MH, Bernatchez L. 2002. PAPA (package for the analysis of parental allocation): a computer program for simulated and real parental allocation. Mol Ecol Notes. 2:191-193.) was investigated under the assumption that only a small proportion of potential breeders contributed to the offspring sample. Inbreeding levels proved to have a large impact on allocation error rate. Consequently, simulations from artificial, unrelated parents may strongly underestimate allocation error, and so, whenever possible, simulations based on the actual parental genotypes should be run. An unexpected and interesting finding was that ambiguity (the highest likelihood is shared by several parental pairs) rates below 10% stood very close to exact allocation error rates (true proportions of wrong allocations). Hence, the ambiguity rate statistic may be viewed as a ready-made indicator of the resolution power of a specific parental allocation run and, if not exceeding 10%, used as an estimate of allocation error rate. It was found that the PAPA simulator, even with few contributing breeders, can be trusted to output reasonably accurate estimates of allocation error as long as those estimates do not exceed 15%. Indeed, most discrepancies between exact and estimated error then stood below 3%. Reproductive success variance had little impact on error estimate discrepancies within the same range. Finally, a (focal set) method was described to correct the estimated family sizes computed directly from parental allocations. Essentially, this method makes use of the detailed structure of the allocation probabilities associated with each parental pair with at least 1 allocated offspring. The allocation probabilities are expressed in matrix form, and the subsequent calculations are run based on standard matrix algebra. On average, this method provided better estimates of family sizes for each investigated combination of parameter values. As the size of offspring samples increased, the corrections improved until a plateau was finally reached. Typically, samples comprising 250, 500, and 1000 offspring would bring corrections in the order of 10-20%, 20-30%, and 30-40%, respectively.

The ecology of sexual dimorphism in size and shape of the freshwater blenny Salaria fluviatilis.

Sexual selection is considered the major cause of sexual dimorphism, but recent observations suggest that natural selection may play a more important role in the evolution of sex differentiation than previously recognized. Therefore, studying the trade-offs between natural selection and sexual selection is crucial to a better understanding of the ecology underlying the evolution of sexual dimorphism. The freshwater blenny Salaria fluviatilis, a fish inhabiting lakes and rivers around the Mediterranean Sea, displays strong sexual dimorphism in size, shape, and behavior (i.e., larger body and head size for males and higher swimming requirements for females during the reproductive period). We tested for differences in sexual dimorphism in size and shape between the populations from lake and river habitats with the goal of identifying the trade-offs between natural and sexual selection that underlie variations in sexual dimorphism in this species. Our results show i) differences in sexual size dimorphism (SSizeD) in accordance to Rensch's rule (i.e., larger individuals in rivers associated with higher SSizeD), and ii) a decrease in shape differentiation between males and females in lake populations. Together, this suggests that the different environmental conditions between lake and river habitats (e.g., resource limitations, predation pressure, water velocity) affect the relative importance of sexual selection in the display of sexual dimorphism within the species. This study highlights the importance of considering the environmental conditions to which populations are exposed to better understand the ecology underlying the evolution of sexual dimorphism.