Summary1. Knowledge of global patterns of biodiversity and regulating variables is indispensable to develop predictive models. 2. The present study used predictive modelling approaches to investigate hypotheses that explain the variation in fish species richness between estuaries over a worldwide spatial extent. Ultimately, such models will allow assessment of future changes in ecosystem structure and function as a result of environmental changes. 3. A comprehensive worldwide data base was compiled of the fish assemblage composition and environmental characteristics of estuaries. Generalized Linear Models were used to quantify how variation in species richness among estuaries is related to historical events, energy dynamics and ecosystem characteristics, while controlling for sampling effects. 4. At the global extent, species richness differed among marine biogeographic realms and continents and increased with mean sea surface temperature, terrestrial net primary productivity and the stability of connectivity with a marine ecosystem (open vs. temporarily open estuaries). At a smaller extent (within a marine biogeographic realm or continent), other characteristics were also important in predicting variation in species richness, with species richness increasing with estuary area and continental shelf width. 5. The results suggest that species richness in an estuary is defined by predictors that are spatially hierarchical. Over the largest spatial extents, species richness is influenced by the broader distributions and habitat use patterns of marine and freshwater species that can colonize estuaries, which are in turn governed by history contingency, energy dynamics and productivity variables. Species richness is also influenced by more regional and local parameters that can further affect the process of community colonization in an estuary including the connectivity of the estuary with the adjacent marine habitat, and, over smaller spatial extents, the size of these habitats. In summary, patterns of species richness in estuaries across large spatial extents seem to reflect from global to local processes acting on community colonization. The importance of considering spatial extent, sampling effects and of combining history and contemporary environmental characteristics when exploring biodiversity is highlighted.

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Aim We present the first global biogeographical regionalization of estuaries, assessing how dispersal limitation and/or environmental filtering mechanisms drive the patterns of fish assemblage composition among and within biogeographical regions. Location Estuaries worldwide.Methods A bootstrapped hierarchical cluster analysis was applied to define biogeographical regions based on pairwise beta diversity (bsim) of fish assemblages among 393 estuaries worldwide. Variables representing dispersal limitation and environmental features were used to disentangle the possible effects of assembly processes acting among and within biogeographical regions, through hierarchical partitioning of variation. Finally, we assessed the contribution of species to beta diversity through a method newly developed in this article.Results Estuaries were grouped into seven major biogeographical regions explained by geographical dispersal variables. Dissimilarity in species composition within each biogeographical region was mainly determined by ecosystemconnectivity dispersal variables, with a minor influence of environmental variables (chiefly sea surface temperature). Marine species contributed highly to beta diversity between estuaries but less than expected by chance, whilst freshwater species had low contributions yet also less than expected. Estuarine (i.e. brackish) and diadromous species had low contributions to beta diversity but higher than expected by chance.Main conclusions Fish assemblage composition in estuaries seems to be driven by dispersal limitation processes both among and within biogeographical regions, with only a minor role of environmental filtering evident within biogeographical regions. These findings contrast with the acknowledged importance of environmental variables as drivers of species richness patterns. Irrespective of biogeographical region, the ecosystem affinity of a species determined its contribution to beta diversity between estuaries. The new method presented here offers new opportunities for assessing the effects of taxonomic and functional aspects on beta diversity (bsim) across taxa and ecosystems, and thus contributes to improving knowledge of assembly processes.

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We hypothesized that cytokines influence luteal angiogenesis in mares, while angiogenic factors themselves can also regulate luteal secretory capacity. Therefore, the purpose of this study was to evaluate the role of cytokines--tumor necrosis factor alpha (TNF), interferon gamma (IFNG) and Fas ligand (FASL)--on in vitro modulation of angiogenic activity and mRNA level of vascular endothelial growth factor A (VEGF), its receptor VEGFR2, thrombospondin 1 (TSP1), and its receptor CD36 in equine corpus luteum (CL) throughout the luteal phase. After treatment, VEGF protein expression was determined in midluteal phase (mid) CL cells. The role of VEGF on regulation of luteal secretory capacity was assessed by progesterone (P(4)) and prostaglandin E(2) (PGE(2)) production and by mRNA levels for steroidogenic enzymes 3-beta-hydroxysteroid dehydrogenase (3betaHSD) and PGE synthase (PGES). In early CL cells, TNF increased angiogenic activity (bovine aortic endothelial cell viability) and VEGF and VEGFR2 mRNA levels and decreased CD36 (real-time PCR relative quantification). In mid-CL cells, TNF increased VEGF mRNA and protein expression (Western blot analysis) and reduced CD36 mRNA levels, while FASL and TNF+IFNG+FASL decreased VEGF protein expression. In late CL cells, TNF and TNF+IFNG+FASL reduced VEGFR2 mRNA, but TNF+IFNG+FASL increased TSP1 and CD36 mRNA. VEGF treatment increased mRNA levels of 3betaHSD and PGES and secretion of P(4) and PGE(2). In conclusion, these findings suggest a novel auto/paracrine action of cytokines, specifically TNF, on the up-regulation of VEGF for angiogenesis stimulation in equine early CL, while at luteolysis, cytokines down-regulated angiogenesis. Additionally, VEGF stimulated P(4) and PGE(2) production, which may be crucial for CL establishment.

Assessing trait–environment relationships is crucial for predicting effects of natural and human‐induced environmental change on biota. We compiled a global database of fish assemblages in estuaries, functional traits of fishes and ecosystem features of estuaries. And we quantified the relative importance of ecosystem features as drivers of patterns of fish functional traits among estuaries worldwide (i.e. drivers of the proportions of fish traits). In addition to biogeographical context, two main environmental gradients regulate traits patterns: firstly temperature, and secondly estuary size and hydrological connectivity of the estuary with the marine ecosystem. Overall, estuaries in colder regions, with larger areas and with higher hydrological connectivity with the marine ecosystem, have higher proportions of marine fish (versus freshwater), macrocarnivores and planktivores (versus omnivores, herbivores and detritivores) and larger fish, with greater maximum depth of distribution and longer lifespan. The observed trait patterns and trait–environment relationships are likely generated by multiple causal processes linked to physiological constraints due to temperature and salinity, size‐dependent biotic interactions, as well as habitat availability and connectivity. Biogeographical context and environmental conditions drive species richness and composition, and present results show that they also drive assemblage traits. The observed trait patterns and trait–environment relationships suggest that assemblage composition is determined by the functional role of species within ecosystems. Conservation strategies should be coordinated globally and ensure protection of an array of estuaries that differ in ecosystem features, even if some of those estuaries do not support high species richness.

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