Genetic characterization and intra-generic relationships of Artemia monica Verrill and A. urmiana Günther

Allozyme variation encoded for by 22 enzyme loci analyzed with starch-gel electrophoresis from samples of gonochoristic Artemia monica and A. urmiana collected from Lake Urmia, Iran and from Mono Lake, CA, USA, respectively, are compared and contrasted with data from representative gonochoristic populations of A. franciscana, A. persimilis and A. salina, as well as diploid, triploid, tetraploid, and pentaploid parthenogenetic forms. Values for parameters measuring degree of genetic variability (number of alleles per locus, proportion of polymorphic loci, and expected heterozygosity) in A. monica and A. urmiana were among the highest in the genus (1.76, 0.46, 0.19, and 2.19, 0.55, and 0.14, respectively). For A. monica, possible factors promoting its high genetic variability are discussed. Mean values for Nei's D between A. monica and A. franciscana were very low (0.09) and consistent with reported cytological and morphological similarities, thus the physiological differentiation which distinguishes the former must involve differences at very few loci. D values from A. urmiana to rest of the gonochoristic species (0.95) are consistent with its taxonomic status and distinctive morphology, while its low genetic distance to the monophyletic Artemia parthenogenetic lineage (mean D = 0.48) suggests a recent common ancestry.     

Global conservation priorities for marine turtles

Where conservation resources are limited and conservation targets are diverse, robust yet flexible priority-setting frameworks are vital. Priority-setting is especially important for geographically widespread species with distinct populations subject to multiple threats that operate on different spatial and temporal scales. Marine turtles are widely distributed and exhibit intra-specific variations in population sizes and trends, as well as reproduction and morphology. However, current global extinction risk assessment frameworks do not assess conservation status of spatially and biologically distinct marine turtle Regional Management Units (RMUs), and thus do not capture variations in population trends, impacts of threats, or necessary conservation actions across individual populations. To address this issue, we developed a new assessment framework that allowed us to evaluate, compare and organize marine turtle RMUs according to status and threats criteria. Because conservation priorities can vary widely (i.e. from avoiding imminent extinction to maintaining long-term monitoring efforts) we developed a “conservation priorities portfolio” system using categories of paired risk and threats scores for all RMUs (n = 58). We performed these assessments and rankings globally, by species, by ocean basin, and by recognized geopolitical bodies to identify patterns in risk, threats, and data gaps at different scales. This process resulted in characterization of risk and threats to all marine turtle RMUs, including identification of the world''s 11 most endangered marine turtle RMUs based on highest risk and threats scores. This system also highlighted important gaps in available information that is crucial for accurate conservation assessments. Overall, this priority-setting framework can provide guidance for research and conservation priorities at multiple relevant scales, and should serve as a model for conservation status assessments and priority-setting for widespread, long-lived taxa.     

Conservation Genetics of the East Pacific Green Turtle (Chelonia mydas) in Michoacan, Mexico

The main continental nesting rookeries of the east Pacific green turtle (EPGT), Chelonia mydas, on the Michoacan (Mexico) coast suffered drastic population declines following intense exploitation in the 1960s--1970s with annual abundance of nesting females plummeting from about 25,000 to an average of about 1400 between 1982 and 2001. Analyses of data from three nDNA microsatellite loci and 400 bp mtDNA control region sequences from a total of 123 nesting females sampled from four Michoacan rookeries found no evidence of population sub-structuring. The recent order of magnitude reduction in the population size shows no apparent impact on genetic diversity in either control region sequences (overall h = 0.48; pi = 0.0036) or microsatellite loci (overall Na = 20.8; Hexp = 0.895). Our estimates of annual effective female population size (Nef; from theta = 2Nemicron) of 1.9-2.3 x 10(3), in spite of being an order of magnitude below historical records, appear to be sufficient to allow recovery of this population without significant loss of genetic diversity. These findings highlight the importance of continued conservation to reverse the decline of this population before it becomes vulnerable to genetic erosion.     

A comparison between total viable count by spread plating and AquaPlak for enumeration of bacteria in water from a shrimp farm

A new system named AquaPlak^® designed for evaluating heterotrophic bacteria and those belonging to the genus Vibrio was compared to the standard method of total viable count by spread plating. Water samples from a shrimp farm were evaluated by the two procedures over a 3-month period. The spread plating technique gave significantly higher viable counts for both Vibrio and heterotrophic bacteria than the AquaPlak^® system.     

Geographic Patterns of Genetic Variation in a Broadly Distributed Marine Vertebrate: New Insights into Loggerhead Turtle Stock Structure from Expanded Mitochondrial DNA Sequences

Previous genetic studies have demonstrated that natal homing shapes the stock structure of marine turtle nesting populations. However, widespread sharing of common haplotypes based on short segments of the mitochondrial control region often limits resolution of the demographic connectivity of populations. Recent studies employing longer control region sequences to resolve haplotype sharing have focused on regional assessments of genetic structure and phylogeography. Here we synthesize available control region sequences for loggerhead turtles from the Mediterranean Sea, Atlantic, and western Indian Ocean basins. These data represent six of the nine globally significant regional management units (RMUs) for the species and include novel sequence data from Brazil, Cape Verde, South Africa and Oman. Genetic tests of differentiation among 42 rookeries represented by short sequences (380 bp haplotypes from 3,486 samples) and 40 rookeries represented by long sequences (∼800 bp haplotypes from 3,434 samples) supported the distinction of the six RMUs analyzed as well as recognition of at least 18 demographically independent management units (MUs) with respect to female natal homing. A total of 59 haplotypes were resolved. These haplotypes belonged to two highly divergent global lineages, with haplogroup I represented primarily by CC-A1, CC-A4, and CC-A11 variants and haplogroup II represented by CC-A2 and derived variants. Geographic distribution patterns of haplogroup II haplotypes and the nested position of CC-A11.6 from Oman among the Atlantic haplotypes invoke recent colonization of the Indian Ocean from the Atlantic for both global lineages. The haplotypes we confirmed for western Indian Ocean RMUs allow reinterpretation of previous mixed stock analysis and further suggest that contemporary migratory connectivity between the Indian and Atlantic Oceans occurs on a broader scale than previously hypothesized. This study represents a valuable model for conducting comprehensive international cooperative data management and research in marine ecology.     

Biosynthesis of heterocyst glycolipids of Anabaena cylindrica

The incorporation of sodium acetate-[1-¹⁴C] into the heterocyst glycolipids of Anabaena cylindrica cultures from 60–234 hr old is reported. Incorporation of radioactivity was maximal in 88 hr old cultures. In 60 hr and 88 hr cultures about 90 % of the radioactivity of the heterocyst glycolipids was found in the non-saponifiable glycolipid fraction, whereas in older cultures this fraction contained only 75 % of the radioactivity. Acid hydrolysis of non-saponifiable heterocyst glycolipid fractions showed that in 60 hr cultures, 81 % of the radioactivity occurs in the lipid moiety, whereas in older cultures a greater proportion (40–53 %) of the radioactivity was found in the sugar residue. The lipid fraction obtained by acid hydrolysis contained a mixture of labelled long chain mono-, di- and trihydric alcohols. In young (60 hr) cultures the primary alcohol fraction was most heavily labelled (57.3 % of the radioactivity in the non-saponifiable glycosides) with much smaller amounts in the diol and triol (8.4 and 15.1 % respectively), whereas in older cultures (234 hr) the primary alcohol (23.6 %) diol (22.5 %) and triol (18.9 %) fractions contained ca equal amounts of radioactivity.     

Príncipe island hawksbills: Genetic isolation of an eastern Atlantic stock

The hawksbill turtle is a critically endangered species that has been extensively exploited for centuries. Príncipe Island off western Africa harbours one of the species' major nesting populations in the eastern Atlantic, as well as hosting year-round foraging aggregations of juveniles, subadults and adult males. To gain insight into the population's genetic structure and relationships with regional stocks, we analysed mitochondrial DNA (mtDNA) sequences of nesting females (N = 9), foraging adult females (N = 11), adult males (N = 32), subadults (N = 15) and juveniles (N = 80). The nesting population was found to be fixed for a single haplotype (EATL), which had been previously reported in both western and eastern Atlantic hawksbill foraging sites but had no known rookery source prior to this study. Thus it is now possible to confirm the westward transoceanic movement by hawksbills originating from Príncipe Island. Our analyses demonstrated that the Príncipe Island nesting colony is genetically distinct from breeding populations in the western Atlantic and is phylogenetically linked with Indo-Pacific hawksbill clades, suggesting that Príncipe Island was most likely colonised by migrants from the Indian Ocean via the Cape of Good Hope in southern Africa. Mixed stock analyses revealed that the eastern Atlantic appears to be the primary foraging area for Príncipe hawksbills (75%) while most of the foraging juveniles in Príncipe waters originate from the Príncipe rookery (84%). Furthermore, the presence of Caribbean haplotypes at low frequencies (< 5%) suggests that eastward transatlantic movements by juveniles to distant foraging and developmental habitats also take place. Depleted hawksbill populations in the eastern Atlantic combined with the low genetic variability and high genetic distinctiveness found in the Príncipe nesting and foraging aggregations with respect to the western Atlantic, underscore the high degree of isolation and vulnerability of this eastern Atlantic stock. These characteristics are highly relevant for the development of effective conservation programmes and highlight the urgent need to consolidate international cooperation across regional boundaries.     

Regional management units for marine turtles: a novel framework for prioritizing conservation and research across multiple scales

Background

Resolving threats to widely distributed marine megafauna requires definition of the geographic distributions of both the threats as well as the population unit(s) of interest. In turn, because individual threats can operate on varying spatial scales, their impacts can affect different segments of a population of the same species. Therefore, integration of multiple tools and techniques — including site-based monitoring, genetic analyses, mark-recapture studies and telemetry — can facilitate robust definitions of population segments at multiple biological and spatial scales to address different management and research challenges.

Methodology/Principal Findings

To address these issues for marine turtles, we collated all available studies on marine turtle biogeography, including nesting sites, population abundances and trends, population genetics, and satellite telemetry. We georeferenced this information to generate separate layers for nesting sites, genetic stocks, and core distributions of population segments of all marine turtle species. We then spatially integrated this information from fine- to coarse-spatial scales to develop nested envelope models, or Regional Management Units (RMUs), for marine turtles globally.

Conclusions/Significance

The RMU framework is a solution to the challenge of how to organize marine turtles into units of protection above the level of nesting populations, but below the level of species, within regional entities that might be on independent evolutionary trajectories. Among many potential applications, RMUs provide a framework for identifying data gaps, assessing high diversity areas for multiple species and genetic stocks, and evaluating conservation status of marine turtles. Furthermore, RMUs allow for identification of geographic barriers to gene flow, and can provide valuable guidance to marine spatial planning initiatives that integrate spatial distributions of protected species and human activities. In addition, the RMU framework — including maps and supporting metadata — will be an iterative, user-driven tool made publicly available in an online application for comments, improvements, download and analysis.     

Dispersal, recruitment and migratory behaviour in a hawksbill sea turtle aggregation

We investigated the dispersal, recruitment and migratory behaviour of the hawksbill sea turtle ( Eretmochelys imbricata ), among different life-history stages and demographic segments of the large hawksbill turtle aggregation at Mona Island, Puerto Rico. There were significant differences in both mitochondrial DNA (mtDNA) haplotype diversity and haplotype frequencies among the adult males, females and juveniles examined, but little evidence for temporal heterogeneity within these same groups sampled across years. Consistent with previous studies and the hypothesis of strong natal homing, there were striking mtDNA haplotype differences between nesting females on Mona Island and nesting females in other major Caribbean rookeries. Breeding males also showed strong, albeit weaker, genetic evidence of natal homing. Overall, Bayesian mixed-stock analysis suggests that Mona Island was the natal rookery for 79% (65–94%) of males in the aggregation. In contrast, the Mona Island rookery accounted for only a small subset of the new juvenile recruits to the foraging grounds or in the population of older juvenile hawksbills turtles on Mona. Instead, both new recruits and the older juvenile hawksbill turtles on Mona more likely recruited from other Caribbean rookeries, suggesting that a mechanism besides natal homing must be influencing recruitment to feeding habitats. The difference in the apparent degree of natal homing behaviour among the different life-history stages of hawksbill turtles at Mona Island underscores the complexity of the species' life-history dynamics and highlights the need for both local and regional conservation efforts.