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.

Environmental Dynamics of Dissolved Black Carbon in Aquatic Ecosystems

Black carbon (BC), the incomplete combustion product from biomass and fossil fuel burning, is ubiquitously found in soils, sediments, ice, water and atmosphere. Because of its polyaromatic molecular characteristic, BC is believed to contribute significantly to the global carbon budget as a slow-cycling, refractory carbon pool. However, the mass balance between global BC generation and accumulation does not match, suggesting a removal mechanism of BC to the active carbon pool, most probable in a dissolved form. The presence of BC in waters as part of the dissolved organic matter (DOM) pool was recently confirmed via ultrahigh resolution mass spectrometry, and dissolved black carbon (DBC), a degradation product of charcoal, was found in marine and coastal environments. However, information on the loadings of DBC in freshwater environments and its global riverine flux from terrestrial systems to the oceans remained unclear. The main objectives of this study were to quantify DBC in diverse aquatic ecosystems and to determine its environmental dynamics. Surface water samples were collected from aquatic environments with a spatially significant global distribution, and DBC concentrations were determined by a chemical oxidation method coupled with HPLC detection. While it was clear that biomass burning was the main sources of BC, the translocation mechanism of BC to the dissolved phase was not well understood. Data from the regional studies and the developed global model revealed a strong positive correlation between DBC and dissolved organic carbon (DOC) dynamics, indicating a co-generation and co-translocation between soil OC and BC. In addition, a DOC-assistant DBC translocation mechanism was identified. Taking advantage of the DOC-DBC correlation model, a global riverine DBC flux to oceans on the order of 26.5 Mt C yr-1 (1 Mt = 1012 g) was determined, accounting for 10.6% of the global DOC flux. The results not only indicated that DOC was an important environmental intermediate for BC transfer and storage, but also provided an estimate of a major missing link in the global BC budget. The ever increasing DBC export caused by global warming will change the marine DOM quality and may have important consequences for carbon cycling in marine ecosystem.

The importance of marine subsidies for terrestrial consumers in coastal Peru

The Peruvian coast is one the best examples of cross-ecosystem food web exchanges, in which resources from one of the richest marine ecosystems subsidize consumers in one of the driest deserts on Earth. Marine subsidies are resources that originate in the marine ecosystem, and that contribute to increase the density of consumers in the recipient ecosystem. I examined the effects of marine subsidies on animal populations in the Peruvian coastal desert. I combined several approaches to study the linkages between marine resources and terrestrial consumers, such as surveying the spatial distribution and estimating the relative abundance of terrestrial consumers, studying the diet of geckos and lizards through stomach content analyses, and examining the desert food web with carbon and nitrogen stable isotope analyses. I found that the distribution and diet of desert consumers were tightly coupled to the availability of marine subsidies. I revealed linkages along two pathways of nutrient fluxes: tidal action that washes ashore macroalgae and cadavers of marine organisms, and animal transport in places where pinnipeds and seabirds congregate for reproduction. In the first pathway, intertidal algivivores made marine resources available to terrestrial consumers by moving between the intertidal and supratidal zone. The relative contribution of terrestrial and algal carbon sources varied among terrestrial consumers, because scorpions assimilated a lower proportion of energy from macroalgae than did geckos and solifuges. In the second pathway, I found that pinniped colonies influenced the diet of desert consumers, and contributed to support large populations of lizards and geckos. By combining field observations, and stomach and stable isotope analyses, I constructed a simplified food web for a large sea lion colony, showing the number of trophic levels that originate from pinniped-derived nutrients. My study demonstrates the enormous importance of marine resources for the diet of desert consumers. The near absence of rainfall along the Peruvian coast promotes an extreme dependence of terrestrial consumers on marine resources, and causes permanent food web effects that are affected by temporal variability in marine productivity, rather then temporal patterns of desert plant growth.

Development of molecular approaches to assess the diversity and abundance of marine nematodes assemblages for assessment of Good Environmental Status of the estuarine and marine ecosystems

In Europe, the concerns with the status of marine ecosystems have increased, and the Marine Directive has as main goal the achievement of Good Environmental Status (GES) of EU marine waters by 2020. Molecular tools are seen as promising and emerging approaches to improve ecosystem monitoring, and have led ecology into a new era, representing perhaps the most source of innovation in marine monitoring techniques. Benthic nematodes are considered ideal organisms to be used as biological indicator of natural and anthropogenic disturbances in aquatic ecosystems underpinning monitoring programmes on the ecological quality of marine ecosystems, very useful to assess the GES of the marine environment. dT-RFLP (directed Terminal-Restriction Fragment Length Polymorphism) allows to assess the diversity of nematode communities, but also allows studying the functioning of the ecosystem, and combined with relative real-time PCR (qPCR), provides a high-throughput semi-quantitative characterization of nematode communities. These characteristics make the two molecular tools good descriptors for the good environmental status assessment. The main aim of this study is to develop and optimize the dT-RFLP and qPCR in Mira estuary (SW coast, Portugal). A molecular phylogenetic analysis of marine and estuarine nematodes is being performed combining morphological and molecular analysis to evaluate the diversity of free-living marine nematodes in Mira estuary. After morphological identification, barcoding of 18S rDNA and COI genes are being determined for each nematode species morphologically identified. So far we generated 40 new sequences belonging to 32 different genus and 17 families, and the study has shown a good degree of concordance between traditional morphology-based identification and DNA sequences. These results will improve the assessment of marine nematode diversity and contribute to a more robust nematode taxonomy. The DNA sequences are being used to develop the dT-RFLP with the ability to easily process large sample numbers (hundreds and thousands), rather than typical of classical taxonomic or low throughput molecular analyses. A preliminary study showed that the digest enzymes used in dT-RFLP for terrestrial assemblages separated poorly the marine nematodes at taxonomic level for functional group analysis. A new digest combination was designed using the software tool DRAT (Directed Terminal Restriction Analysis Tool) to distinguished marine nematode taxa. Several solutions were provided by DRAT and tested empirically to select the solution that cuts most efficiently. A combination of three enzymes and a single digest showed to be the best solution to separate the different clusters. Parallel to this, another tool is being developed to estimate the population size (qPCR). An improvement in qPCR estimation of gene copy number using an artificial reference is being performed for marine nematodes communities to quantify the abundance. Once developed, it is proposed to validate both methodologies by determining the spatial and temporal variability of benthic nematodes assemblages across different environments. The application of these high-throughput molecular approaches for benthic nematodes will improve sample throughput and their implementation more efficient and faster as indicator of ecological status of marine ecosystems.

Chitosan Nanoparticles Loaded The Herbicide Paraquat: The Influence Of The Aquatic Humic Substances On The Colloidal Stability And Toxicity.

Polymeric nanoparticles have been developed for several applications, among them as carrier system of pesticides. However, few studies have investigated the fate of these materials in the environment in relation to colloidal stability and toxicity. In nature, humic substances are the main agents responsible for complexation with metals and organic compounds, as well as responsible for the dynamics of these nanoparticles in aquatic and terrestrial environments. In this context, the evaluation of the influence of aquatic humic substances (AHS) on the colloidal stability and toxicity of polymeric nanoparticles of chitosan/tripolyphosphate with or without paraquat was performed. In this study, the nanoparticles were prepared by the ionic gelation method and characterized by size distribution measurements (DLS and NTA), zeta potential, infrared and fluorescence spectroscopy. Allium cepa genotoxicity studies and ecotoxicity assays with the alga Pseudokirchneriella subcapitata were used to investigate the effect of aquatic humic substances (AHS) on the toxicity of this delivery system. No changes were observed in the physical-chemical stability of the nanoparticles due to the presence of AHS using DLS and NTA techniques. However some evidence of interaction between the nanoparticles and AHS was observed by infrared and fluorescence spectroscopies. The ecotoxicity and genotoxicity assays showed that humic substances can decrease the toxic effects of nanoparticles containing paraquat. These results are interesting because they are important for understanding the interaction of these nanostructured carrier systems with species present in aquatic ecosystems such as humic substances, and in this way, opening new perspectives for studies on the dynamics of these carrier systems in the ecosystem.

Convergence Of Soil Nitrogen Isotopes Across Global Climate Gradients.

Quantifying global patterns of terrestrial nitrogen (N) cycling is central to predicting future patterns of primary productivity, carbon sequestration, nutrient fluxes to aquatic systems, and climate forcing. With limited direct measures of soil N cycling at the global scale, syntheses of the (15)N:(14)N ratio of soil organic matter across climate gradients provide key insights into understanding global patterns of N cycling. In synthesizing data from over 6000 soil samples, we show strong global relationships among soil N isotopes, mean annual temperature (MAT), mean annual precipitation (MAP), and the concentrations of organic carbon and clay in soil. In both hot ecosystems and dry ecosystems, soil organic matter was more enriched in (15)N than in corresponding cold ecosystems or wet ecosystems. Below a MAT of 9.8°C, soil δ(15)N was invariant with MAT. At the global scale, soil organic C concentrations also declined with increasing MAT and decreasing MAP. After standardizing for variation among mineral soils in soil C and clay concentrations, soil δ(15)N showed no consistent trends across global climate and latitudinal gradients. Our analyses could place new constraints on interpretations of patterns of ecosystem N cycling and global budgets of gaseous N loss.

Restricted-Range Fishes and the Conservation of Brazilian Freshwaters

Background: Freshwaters are the most threatened ecosystems on earth. Although recent assessments provide data on global priority regions for freshwater conservation, local scale priorities remain unknown. Refining the scale of global biodiversity assessments (both at terrestrial and freshwater realms) and translating these into conservation priorities on the ground remains a major challenge to biodiversity science, and depends directly on species occurrence data of high taxonomic and geographic resolution. Brazil harbors the richest freshwater ichthyofauna in the world, but knowledge on endemic areas and conservation in Brazilian rivers is still scarce. Methodology/Principal Findings: Using data on environmental threats and revised species distribution data we detect and delineate 540 small watershed areas harboring 819 restricted-range fishes in Brazil. Many of these areas are already highly threatened, as 159 (29%) watersheds have lost more than 70% of their original vegetation cover, and only 141 (26%) show significant overlap with formally protected areas or indigenous lands. We detected 220 (40%) critical watersheds overlapping hydroelectric dams or showing both poor formal protection and widespread habitat loss; these sites harbor 344 endemic fish species that may face extinction if no conservation action is in place in the near future. Conclusions/Significance: We provide the first analysis of site-scale conservation priorities in the richest freshwater ecosystems of the globe. Our results corroborate the hypothesis that freshwater biodiversity has been neglected in former conservation assessments. The study provides a simple and straightforward method for detecting freshwater priority areas based on endemism and threat, and represents a starting point for integrating freshwater and terrestrial conservation in representative and biogeographically consistent site-scale conservation strategies, that may be scaled-up following naturally linked drainage systems. Proper management (e. g. forestry code enforcement, landscape planning) and conservation (e. g. formal protection) of the 540 watersheds detected herein will be decisive in avoiding species extinction in the richest aquatic ecosystems on the planet.

Gaseous and fluvial carbon export from an Amazon forest watershed

The transfer of carbon (C) from Amazon forests to aquatic ecosystems as CO(2) supersaturated in groundwater that outgases to the atmosphere after it reaches small streams has been postulated to be an important component of terrestrial ecosystem C budgets. We measured C losses as soil respiration and methane (CH(4)) flux, direct CO(2) and CH(4) fluxes from the stream surface and fluvial export of dissolved inorganic C (DIC), dissolved organic C (DOC), and particulate C over an annual hydrologic cycle from a 1,319-ha forested Amazon perennial first-order headwater watershed at Tanguro Ranch in the southern Amazon state of Mato Grosso. Stream pCO(2) concentrations ranged from 6,491 to 14,976 mu atm and directly-measured stream CO(2) outgassing flux was 5,994 +/- A 677 g C m(-2) y(-1) of stream surface. Stream pCH(4) concentrations ranged from 291 to 438 mu atm and measured stream CH(4) outgassing flux was 987 +/- A 221 g C m(-2) y(-1). Despite high flux rates from the stream surface, the small area of stream itself (970 m(2), or 0.007% of watershed area) led to small directly-measured annual fluxes of CO(2) (0.44 +/- A 0.05 g C m(2) y(-1)) and CH(4) (0.07 +/- A 0.02 g C m(2) y(-1)) per unit watershed land area. Measured fluvial export of DIC (0.78 +/- A 0.04 g C m(-2) y(-1)), DOC (0.16 +/- A 0.03 g C m(-2) y(-1)) and coarse plus fine particulate C (0.001 +/- A 0.001 g C m(-2) y(-1)) per unit watershed land area were also small. However, stream discharge accounted for only 12% of the modeled annual watershed water output because deep groundwater flows dominated total runoff from the watershed. When C in this bypassing groundwater was included, total watershed export was 10.83 g C m(-2) y(-1) as CO(2) outgassing, 11.29 g C m(-2) y(-1) as fluvial DIC and 0.64 g C m(-2) y(-1) as fluvial DOC. Outgassing fluxes were somewhat lower than the 40-50 g C m(-2) y(-1) reported from other Amazon watersheds and may result in part from lower annual rainfall at Tanguro. Total stream-associated gaseous C losses were two orders of magnitude less than soil respiration (696 +/- A 147 g C m(-2) y(-1)), but total losses of C transported by water comprised up to about 20% of the +/- A 150 g C m(-2) (+/- 1.5 Mg C ha(-1)) that is exchanged annually across Amazon tropical forest canopies.

Spatial variation in the strength of mutualism between a jumping spider and a terrestrial bromeliad: Evidence from the stable isotope N-15

Psecas chapoda, a neotropical jumping spider strictly associated with the terrestrial bromeliad Bromelia balansae in cerrados and semi-deciduous forests in South America, effectively contributes to plant nutrition and growth. In this study, our goal was to investigate if spider density caused spatial variations in the strength of this spider-plant mutualism. We found a positive significant relationship between spider density and delta N-15 values for bromeliad leaves in different forest fragments. Open grassland Bromeliads were associated with spiders and had higher delta N-15 values compared to forest bromeliads. Although forest bromeliads had no association with spiders their total N concentrations were higher. These results suggest that bromeliad nutrition is likely more litter-based in forests and more spider-based in open grasslands. This study is one of the few to show nutrient provisioning and conditionality in a spider-plant system. (c) 2008 Elsevier Masson SAS. All rights reserved.

Soil mineralogy of mangrove forests from the state of Sao Paulo, Southeastern Brazil

The minerals of the clay fraction in estuarine plains are mainly detrital being a mixture of marine and continental sediments, but can also be authigenic. Because of the importance of mangrove ecosystems in tropical estuarine areas and the relatively few existing studies of the mineralogical composition of soils in these environments, the aim of this study was to determine the mineralogical assemblage and identify potential contrasts along the coast of the State of Sao Paulo. Soils from I I mangroves distributed along the coastal plain of the State of Sao Paulo were sampled at depths of 0 to 20 and 60 to 80 cm, and samples of suspended sediments from the Ribeira do Iguape River were collected for analysis. Mineralogical analyses were performed on the clay and silt fractions by x-ray diffraction (XRD) and transmission electron microscopy, and fresh soil samples were analyzed by scanning electron microscopy-energy dispersive spectrometry and suspended sediments by XRD. The silt fraction contained quartz, feldspars, gibbsite, kaolinite, illite, and vermiculite, and the clay fraction contained smectite, kaolinite, illite, gibbsite, quartz, and feldspars. Locally, vermiculite, biotite, anatase, halloysite, and goethite may occur because of recent transport of sediments to the system. Pyrite was identified in fresh samples. The allochthonous minerals found either were terrestrial and transported by rivers or had originated from the continental platform by past transgressive events. We suggest that the neoformation of smectite and kaolinite occurs in mangrove soils. Different geomorphological settings along the Sao Paulo coast appear to regulate mineral distribution in mangrove soils.

Do Terrestrial Tank Bromeliads in Brazil Create Safe Sites for Palm Establishment or Act as Natural Traps for Its Dispersed Seeds ?

This study shows for the first time that terrestrial tank bromeliads from Brazilian restinga can act as natural traps for dispersed palm Euterpe edulis seeds. Such bromeliads, which are shade intolerant, gain benefits by limiting palm recruitment since they hinder canopy formation and, consequently, increase luminosity over its aggregates.

Microcystins in South American aquatic ecosystems: Occurrence, toxicity and toxicological assays

The acute poisoning of chronic renal patients during hemodialysis sessions in 1996 in Caruaru City (Pernambuco State, Brazil) stimulated an intensive search for the cause of this severe complication. This search culminated in the identification of microcystins (MC), hepatotoxic cyclic heptapeptides produced by cyanobacteria, as the causative agents. More than ten years later, additional research data provides us with a better understanding of the factors related to cyanobacterial bloom occurrence and production of MC in Brazil and other South American countries. The contamination of water bodies and formation of toxic blooms remains a very serious concern, especially in countries in which surface water is used as the main source for human consumption. The purpose of this review is to highlight the discoveries of the past 15 years that have brought South American researchers to their current level of understanding of toxic cyanobacteria species and that have contributed to their knowledge of factors related to MC production, mechanisms of action and consequences for human health and the environment. (C) 2010 Elsevier Ltd. All rights reserved.

The N-15 natural abundance (delta N-15) of ecosystem samples reflects measures of water availability

We assembled a globally-derived data set for site-averaged foliar delta(15)N, the delta(15)N of whole surface mineral soil and corresponding site factors (mean annual rainfall and temperature, latitude, altitude and soil pH). The delta(15)N of whole soil was related to all of the site variables (including foliar delta(15)N) except altitude and, when regressed on latitude and rainfall, provided the best model of these data, accounting for 49% of the variation in whole soil delta(15)N. As single linear regressions, site-averaged foliar delta(15)N was more strongly related to rainfall than was whole soil delta(15)N. A smaller data set showed similar, negative correlations between whole soil delta(15)N, site-averaged foliar delta(15)N and soil moisture variations during a single growing season. The negative correlation between water availability (measured here by rainfall and temperature) and soil or plant delta(15)N fails at the landscape scale, where wet spots are delta(15)N-enriched relative to their drier surroundings. Here we present global and seasonal data, postulate a proximate mechanism for the overall relationship between water availability and ecosystem delta(15)N and, newly, a mechanism accounting for the highly delta(15)N-depleted values found in the foliage and soils of many wet/cold ecosystems. These hypotheses are complemented by documentation of the present gaps in knowledge, suggesting lines of research which will provide new insights into terrestrial N-cycling. Our conclusions are consistent with those of Austin and Vitousek (1998) that foliar (and soil) delta(15)N appear to be related to the residence time of whole ecosystem N.

The last glacial cycle from the humid tropics of northeastern Australia: comparison of a terrestrial and a marine record

A detailed pollen record from the Ocean Drilling Program Site 820 core, located on the upper part of the continental slope off the coast of northeast Queensland, was constructed to compare with the existing pollen record from Lynch's Crater on the adjacent Atherton Tableland and allow the production of a regional picture of vegetation and environmental change through the last glacial cycle. Some broad similarities in patterns of vegetation change are revealed, despite the differences between sites and their pollen catchments, which can be related largely to global climate and sea-level changes. The original estimated time scale of the Lynch's Crater record is largely confirmed from comparison with the more thoroughly dated ODP record. Conversely, the Lynch's Crater pollen record has assisted in dating problematic parts of the ODP record. In contrast to Lynch's Crater, which reveals a sharp and sustained reduction in drier araucarian forest around 38,000 yrs BP, considered to have been the result of burning by Aboriginal people, the ODP record indicates, most likely, a stepwise reduction, dating from 140,000 yrs BP or beyond. The earliest reduction shows lack of a clear connection between Araucaria decline and increased burning and suggests that people may not have been involved at this stage. However, a further decline in araucarian forest, possibly around 45,000 yrs BP, which has a more substantial environmental impact and is not related to a time of major climate change, is likely, at least partially, the result of human burning. The suggestion, from the ODP core oxygen isotope record, of a regional sea-surface temperature increase of around 4 degrees C between about 400,000 and 250,000 yrs BP, may have had some influence on the overall decline in Araucaria and its replacement by sclerophyll vegetation. (C) 2000 Elsevier Science B.V. All rights reserved.

Unifying principles in terrestrial locomotion: Do hopping Australian marsupials fit in?

Mammalian terrestrial locomotion has many unifying principles. However, the Macropodoidea are a particularly interesting group that exhibit a number of significant deviations from the principles that seem to apply to other mammals. While the properties of materials that comprise the musculoskeletal system of mammals are similar, evidence suggests that tendon properties in macropodoid marsupials may be size or function dependent, in contrast to the situation in placental mammals. Postural differences related to hopping versus running have a dramatic effect on the scaling of the pelvic limb musculoskeletal system. Ratios of muscle fibre to tendon cross-sectional areas for ankle extensors and digital flexors scale with positive allometry in all mammals, but exponents are significantly higher in macropods. Tendon safety factors decline with increasing body mass in mammals, with eutherians at risk of ankle extensor tendon rupture at a body mass of about 150 kg, whereas kangaroos encounter similar problems at a body mass of approximately 35 kg. Tendon strength appears to limit locomotor performance in these animals. Elastic strain energy storage in tendons is mass dependent in all mammals, but exponents are significantly larger in macropodid. Tibial stresses may scale with positive allometry in kangaroos, which result in lower bone safety factors in macropods compared to eutherian mammals.