Genetic conectivity patterns in Holothuria Mammata considering different spatial scales

Dissertação de Mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Phylogeography of the Atlanto-Mediterranean sea cucumber Holothuria (Holothuria) mammata: the combined effects of historical processes and current oceanographical pattern

We assessed the genetic structure of populations of the widely distributed sea cucumber Holothuria (Holothuria) mammata Grube, 1840, and investigated the effects of marine barriers to gene flow and historical processes. Several potential genetic breaks were considered, which would separate the Atlantic and Mediterranean basins, the isolated Macaronesian Islands from the other locations analysed, and the Western Mediterranean and Aegean Sea (Eastern Mediterranean). We analysed mitochondrial 16S and COI gene sequences from 177 individuals from four Atlantic locations and four Mediterranean locations. Haplotype diversity was high (H = 0.9307 for 16S and 0.9203 for COI), and the haplotypes were closely related (p = 0.0058 for 16S and 0.0071 for COI). The lowest genetic diversities were found in the Aegean Sea population. Our results showed that the COI gene was more variable and more useful for the detection of population structure than the 16S gene. The distribution of mtDNA haplotypes, the pairwise FST values and the results of exact tests and AMOVA revealed: (i) a significant genetic break between the population in the Aegean Sea and those in the other locations, as supported by both mitochondrial genes, and (ii) weak differentiation of the Canary and Azores Islands from the other populations; however, the populations from the Macaronesian Islands, Algarve and West Mediterranean could be considered to be a panmictic metapopulation. Isolation by distance was not identified in H. (H.) mammata. Historical events behind the observed findings, together with the current oceanographic patterns, were proposed and discussed as the main factors that determine the population structure and genetic signature of H. (H.) mammata

Assessment of sea cucumber populations from the Aegean Sea (Turkey): first insights to sustainable management of new fisheries

Sea cucumber stocks have been overfished in many countries. As a consequence, several species (Holothuria polii, Holothuria tubulosa and Holothuria mammata) are now caught in Turkish waters without adequate knowledge on their biology and ecology. Here, we address their morphometry, relationships among gutted length and weight, population dynamics, temporal evolution of catches, and we provide the first insights about technical aspects of their fisheries. The largest size classes of H. polii are missing from our sampling collection, possibly due to the heavy fishery pressure on this species. Significant differences in the eviscerated length and weight were found among the Turkish sampled localities for H. polii and H. tubulosa, respectively. These differences could be explained by higher food availability in some areas and/or differential fishery pressure. The size and weight of H. tubulosa specimens were smaller than those registered for the same species in Greek waters, where this species is not fished. All the studied species showed allometric growth. In the last two years, the sea cucumber fishery in Turkey has been increasing rapidly, reaching a total production of ca. 555 000 kg in 2012 (80% H. polii and 20% H. tubulosa plus H. mammata). For a correct management of these species, we recommend: 1) the reestablishment of species-specific closed fishery season according to the specific reproductive cycle; 2) the assessment of the exploited stocks from the Northern Turkish coasts with estimates of recovery time of their populations; 3) the reduction of fishery efforts, mainly on H. polii and H. tubulosa and 4) the establishment of protected areas (where sea cucumber fisheries are forbidden) to conserve healthy populations which will favour the recruitment on nearby areas.

Genetic diversity and connectivity remain high in Holothuria polii (Delle Chiaje 1823) across a coastal lagoon-open sea environmental gradient

Coastal lagoons represent habitats with widely heterogeneous environmental conditions, particularly as regards salinity and temperature,which fluctuate in both space and time. These characteristics suggest that physical and ecological factors could contribute to the genetic divergence among populations occurring in coastal lagoon and opencoast environments. This study investigates the genetic structure of Holothuria polii at a micro-geographic scale across theMar Menor coastal lagoon and nearbymarine areas, estimating the mitochondrial DNA variation in two gene fragments, cytochrome oxidase I (COI) and 16S rRNA (16S). Dataset of mitochondrial sequences was also used to test the influence of environmental differences between coastal lagoon andmarine waters on population genetic structure. All sampled locations exhibited high levels of haplotype diversity and low values of nucleotide diversity. Both genes showed contrasting signals of genetic differentiation (non-significant differences using COI and slight differences using 16S, which could due to different mutation rates or to differential number of exclusive haplotypes. We detected an excess of recent mutations and exclusive haplotypes, which can be generated as a result of population growth. However, selective processes can be also acting on the gene markers used; highly significant generalized additive models have been obtained considering genetic data from16S gene and independent variables such as temperature and salinity.

West versus East Mediterranean Sea: origin and genetic differentiation of the sea cucumber Holothuria polii

We studied the genetic structure of the sea cucumber Holothuria (Roweothuria) polii (Delle Chiaje 1823) by analysing the mitochondrial DNA variation in two fragments of cytochrome oxidase I (COI) and 16S genes. Individuals were collected in seven locations along the Mediterranean Sea, which cover a wide range of the species distribution. We found high haplotype diversity for COI and moderate diversity for 16S, and low nucleotide diversity for both genes. Our results for the COI gene showed many recent and exclusive haplotypes with few mutational changes, suggesting recent or ongoing population expansion. The Western and Eastern Mediterranean populations exhibited slight but significant genetic differentiation (COI gene) with higher genetic diversity in the East. The most ancient haplotype was not present in the westernmost sampling location (SE Spain). The oldest expansion time was observed in Turkey, corresponding to mid-Pleistocene. Turkey had also the highest genetic diversity (number of total and exclusive haplotypes, polymorphisms, haplotype and nucleotide diversity). This suggests that this region could be the origin of the subsequent colonizations through the Mediterranean Sea, a hypothesis that should be assessed with nuclear markers in future research.

Spatial distribution patterns and movements of Holothuria arguinensis in the Ria Formosa (Portugal)

Holothurian populations are under pressure worldwide because of increasing demand for beche-de-mer, mainly for Asian consumption. Importations to this area from new temperate fishing grounds provide economic opportunities but also raise concerns regarding future over-exploitation. Studies on the habitat preferences and movements of sea cucumbers are important for the management of sea cucumber stocks and sizing of no-take zones, but information on the ecology and behavior of temperate sea cucumbers is scarce. This study describes the small-scale distribution and movement patterns of Holothuria arguinensis in the intertidal zone of the Ria Formosa national park (Portugal).Mark/recapture studieswere performed to record theirmovements over time on different habitats (sand and seagrass). H. arguinensis preferred seagrass habitats and did not show a size or life stage-related spatial segregation. Its density was 563 ind. ha−1 and mean movement speed was 10 m per day. Movement speed did not differ between habitats and the direction of movement was offshore during the day and shoreward during the night. Median home range size was 35 m2 and overlap among home ranges was 84%. H. arguinensis' high abundance, close association with seagrass and easy catchability in the intertidal zone, indicate the importance of including intertidal lagoons in future studies on temperate sea cucumber ecology since those systems might require different management strategies than fully submerged habitats.

Genetic diversity across geographical scales in marine coastal ecosystems: Holothuria arguinensis a model species

Coastal lagoons are considered one of the most productive areas of our planet harboring a large variety of habitats. Their transitional character, between terrestrial and marine environments, creates a very particular ecosystem with important variations of its environmental conditions. The organisms that are able to survive on these ecosystems frequently experience strong selective pressures and constrictions to gene flowwith marine populations, which could contribute to genetic divergence among populations inhabiting coastal lagoon and marine environments. Therefore, the main aims of this study are to asses the genetic diversity and population structure of Holothuria arguinensis across geographical ranges, to test the hypothesis of coastal lagoons as hotspots of genetic diversity in the Ria Formosa lagoon, and to determine the role of exporting standing genetic variation from the lagoon to open sea and their implications to recent geographical expansion events. To reach these objectives, we investigate the genetic structure of H. arguinensis using two mitochondrial DNA markers (COI and 16S) at different spatial scales: i) small, inside Ria Formosa coastal lagoon, South Portugal; 2) large, including most of the geographical distribution of this species (South and Western Portuguese coast and Canary islands); these results will allow us to compare the genetic diversity of lagoonal and marine populations of H. arguinensis. On this framework, its recent geographical expansion events, recorded by Rodrigues (2012) and González-Wangüemert and Borrero-Pérez (2012), will be analyzed considering the potential contribution from lagoonal genetic pool. Non-significant genetic structure and high haplotypic diversity were found inside the Ria Formosa coastal lagoon. Both genes were unable to detect significant genetic differentiation among lagoonal and marine localities, suggesting a high rate of gene flow. The results supported our hypotheses that coastal lagoons are not only acting as hotspots of genetic diversity, but also contributing for the genetic variability of the species, working as a source of new haplotypes and enhancing adaptation to the high variable conditions. Different genetic patterns of colonization were found on H. arguinensis, but they must be studied more deeply.

Effects of fishery protection on biometry and genetic structure of two target sea cucumber species from the Mediterranean Sea

Sea cucumber fisheries are now occurring in most of the tropical areas of the world, having expanded from its origin in the central Indo-Pacific. Due to the overexploitation of these resources and the increasing demand from Asian countries, new target species from Mediterranean Sea and northeastern Atlantic Ocean are being caught. The fishery effects on biometry and genetic structure of two target species (Holothuria polii and H. tubulosa) from Turkey, were assessed. The heaviest and largest individuals of H. polii were found into the non-fishery area of Kusadasi, also showing the highest genetic diversity. Similar pattern was detected in H. tubulosa, but only the weight was significantly higher in the protected area. However, the observed differences on the fishery effects between species, could be explained considering the different percentage of catches (80% for H. polii and 20% for H. tubulosa).

In two waters: contemporary evolution of lagoonal and marine white seabream (Diplodus sargus) populations

Brackish water ecosystems are often exposed to wide variations in environmental variables, including temperature and salinity, which may cause strong selective pressures on organisms modifying the genetic patterns of species. The aim of this work was to test whether there is a ‘divergence-with-gene flow’ in coastal lagoon populations of white seabream (Diplodus sargus) (Ria Formosa, S Portugal and Mar Menor, SE Spain) respect to four marine populations, by using partial sequences of cyt b mitochondrial gene and information from nine microsatellite loci. Genetic diversity was highest in both coastal lagoons (Mar Menor and Ria Formosa) considering mitochondrial and nuclear markers. Although some of FST population pairwise comparisons were not significant, analyses of molecular variance (AMOVAs) detected differences between groups (coastal lagoon and marine) close to significance. Also, only two haplotypes (Cytb-17 and Cytb-18) were detected in both coastal lagoon sampling sites and these localities (Mar Menor and Ria Formosa) showed the highest number of singletons, some of them with a high number of mutations, as has been already described for other Mar Menor populations (Pomatochistus marmoratus and Holothuria polii). Also, several tests detected significant positive and balancing selection considering mtDNA and microsatellite data. These data support the hypothesis of selection as one of the drivers of the genetic differences found between coastal lagoon and marine populations. The life strategy adopted by Diplodus sargus in coastal lagoons allows it to decrease its mortality rate and improve the heritability of its genes. Also, the increase time spent in coastal lagoons with different temperatures and salinities favours the fitness selection and the maintenance of exclusive haplotypes and genotypes in coastal lagoon inhabitants favouring the ‘divergence-with-gene-flow’.