Sofia Henriques

Gene transcription of TLR2, TLR4, LPS ligands and prostaglandin synthesis enzymes are up-regulated in canine uteri with cystic endometrial hyperplasia–pyometra complex

Escherichia coli (E. coli) is the most frequent bacterium isolated in cases of cystic endometrial hyperplasia-pyometra complex, the most frequent endometrial disorder in the bitch. Toll-like receptors (TLRs) play an essential role in the innate immune system. The aim of this study was to compare transcription of genes encoding TLR2, TLR4 and LPS ligands (CD14, MD-2, LBP), prostaglandin synthesis enzymes (COX1, COX2, PGES1 and PGFS), and to compare COX1 and COX2 protein expression and PGE(2) and PGF(2alpha) endometrial content in the endometrium of canine diestrous uteri with (n=7) or without (n=7) pyometra. All cases of pyometra were hyperplastic and E. coli was the only isolated bacteria, while diestrous normal uteri did not present signs of cystic endometrial hyperplasia and were negative for bacteriology. Except for COX1, transcription of all genes was significantly higher in pyometra than in normal endometria. COX1 protein was observed in both normal and pyometra uteri, but COX2 protein was only present in pyometra cases. Endometrial PGE(2) and PGF(2alpha) content were significantly higher in pyometra than in normal diestrous endometria. In conclusion, data obtained in this study provides evidence that pyometra-isolated E. coli induces the up-regulation of TLR2 and TLR4 genes in the canine diestrous endometrium. This up-regulation, which is probably the result of the stimulation by LPS and lipoprotein E. coli constituents, leads to the endometrial up-regulation of PG synthesis genes. This, in turn, results in a higher endometrial concentration of PGE(2) and PGF(2alpha), which may further regulate the local inflammatory response.

Global patterns and predictors of fish species richness in estuaries.

1. 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.

Development of a fish-based multimetric index to assess the ecological quality of marine habitats: the Marine Fish Community Index

In this paper the Marine Fish Community Index (MFCI) for the assessment of ecological status of marine environment is proposed. The MFCI was divided into 4 typologies: Rocky subtidal; shallow, intermediate and deep soft-bottoms. Based on the typical community associated to each typology and the DPSIR analysis performed, a set of metrics were selected and tested through a multiple correlation matrix (Pearsons coefficient) and the core ones included in the index. The MFCI was applied in all typologies and the scores obtained with each metric were analyzed. In order to test the robustness of the MFCI the final ecological value of each zone was recalculated by removing successively one metric at a time. The MFCI showed a sensitive and robust response in the ecological status assessment. Since it incorporates both functional and structural community information, the MFCI can be useful in the context of the Marine Strategy Framework Directive as well as in other contexts of conservation and sustainable management of the marine environment.

Efficacy of adapted estuarine fish-based multimetric indices as tools for evaluating ecological status of the marine environment

The assessment of ecological status of marine fish communities required by the marine strategy framework directive (MSFD) emphasises the need for fish-based ecological indices in marine waters. In this study we adapt five estuarine multimetric indices to the marine environment and apply them in three types of substrates, analysing the metrics responsible for the obtained patterns of ecological status. The results show inefficiency of the community degradation index (CDI) and the biological health index (BHI) in ecological status assessment and disagreement between the estuarine biotic integrity index (EBI), the estuarine fish community index (EFCI) and the transitional fish classification index (TFCI). Analysis of individual metrics suggests lack of representativeness and consideration for the particularities of each substrates typical fish communities. None of the tested indices were efficient on the marine environment, urging the need for new marine indices that account for differences between types of substrate and depth.

Structural and functional trends indicate fishing pressure on marine fish assemblages

pressure on marine fish assemblages Sofia Henriques*, Miguel P. Pais, Rita P. Vasconcelos, Alberto Murta, Manuela Azevedo, Maria J. Costa and Henrique N. Cabral Centro de Oceanografia, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal; Instituto Português do Mar e da Atmosfera, Av. de Bras ılia, 1449-006 Lisboa, Portugal; and Departamento de Biologia Animal, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal

Current limitations of global conservation to protect higher vulnerability and lower resilience fish species

Estuaries are threatened by intense and continuously increasing human activities. Here we estimated the sensitivity of fish assemblages in a set of estuaries distributed worldwide (based on species vulnerability and resilience), and the exposure to cumulative stressors and coverage by protected areas in and around those estuaries (from marine, estuarine and freshwater ecosystems, due to their connectivity). Vulnerability and resilience of estuarine fish assemblages were not evenly distributed globally and were driven by environmental features. Exposure to pressures and extent of protection were also not evenly distributed worldwide. Assemblages with more vulnerable and less resilient species were associated with estuaries in higher latitudes (in particular Europe), and with higher connectivity with the marine ecosystem, moreover such estuaries were generally under high intensity of pressures but with no concomitant increase in protection. Current conservation schemes pay little attention to species traits, despite their role in maintaining ecosystem functioning and stability. Results emphasize that conservation is weakly related with the global distribution of sensitive fish species in sampled estuaries, and this shortcoming is aggravated by their association with highly pressured locations, which appeals for changes in the global conservation strategy (namely towards estuaries in temperate regions and highly connected with marine ecosystems).

Oestrous cycle-dependent expression of Fas and Bcl2 family gene products in normal canine endometrium

During the oestrous cycle canine endometrium undergoes cyclical cellular proliferation, apoptosis and differentiation. To study the regulation of endometrial apoptosis and proliferation events the expression of apoptosis-related genes was analysed by real-time polymerase chain reaction and cellular expression of their proteins was identified through immunohistochemistry. Cellular apoptosis and proliferation events were detected by TdT-mediated dUTP-biotin nick end labeling (TUNEL) and proliferation marker Ki67 immunostaining, respectively. The highest proliferative index was observed in the follicular phase (all endometrial cellular components) and at early dioestrus (basal glands). This was associated with a low apoptotic index and a strong expression of anti- (Bcl2) and pro-apoptotic proteins (Fas, FasL, Bax). Subsequently (Days 11-45 of dioestrus), basal glandular epithelium experienced the highest apoptotic index, coincidental with a decrease of Bcl2 expression and a low ratio of Bcl2/Bax transcription. An increase in the apoptotic index of crypts, stromal and endothelial cells was observed at late dioestrus and the beginning of anoestrus. These results indicate that pro- and anti-apoptotic proteins regulate the balance between cell proliferation and death in the canine endometrium during the oestrous cycle. High Bcl2 expression in both the follicular and early dioestrous phases stimulate glandular proliferation and prevent apoptosis but, in the non-pregnant uterus, a decrease in Bcl2 expression together with an increase in pro-apoptotic proteins induces apoptosis of basal glandular epithelium cells.

Ichthyofauna of the Selvagens Islands. Do small coastal areas show high species richness in the northeastern Atlantic

Abstract The Selvagens Islands are located in the northeastern Atlantic between the Canary Islands and Madeira Island. As a result of their small size, remote location and harsh sea conditions only a few studies have been conducted to describe their marine species diversity. We were able to identify 29 new coastal fish species, an increase of 33% in the ichthyofauna described for these islands (n = 88). There is a prevalence of species with tropical affinities and only 2.3% (n = 2) are endemic to Macaronesia. Considered a stepping-stone colonization vector from the nearest continental shore, as proposed by other authors for this region, the Selvagens Islands host 34.1% of the ichthyofauna described for the much larger Canary Islands (nspecies = 258, submerged area nSelvagensIs. = 2.3%) and 47.3% of the ichthyofauna described for the more distantly located Madeira Island (nspecies = 186, submerged area nSelvagensIs. = 17.9%). Interestingly, 6.8% (n = 6) of the species failed to bridge the gap between the Selvagens Islands and Madeira Island. Data collected so far showed no trend toward an increasing number of species with high dispersal capability. The Selvagens Islands are an example of a high coastal species diversity occurring even in very small areas of the northeastern Atlantic Ocean.