Percorrer por autor "Frade, Pedro R."
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- Confocal laser scanning microscopy reveals species-specific differences in distribution of fluorescent proteins in coral tissuesPublication . Marchioro, Giulia M.; Coelho, David; Bouderlique, Thibault; Abed-Navandi, Daniel; Schagerl, Michael; D’Angelo, Cecilia; Kruckenhauser, Luise; Adameyko, Igor; Frade, Pedro R.Reef-building corals have a variety of green fluorescent protein (GFP)-like proteins, also known as fluorescent proteins (FPs). These proteins have broad spectral properties covering most of the visible spectrum, with fluorophores fluorescing from cyan to red. However, the role of FPs is still a topic of debate and requires further investigation, particularly in the direction of mapping these FPs within the coral tissue and describing their cell- and tissue-level distributions. This study applied confocal laser scanning microscopy (CLSM) to investigate species-specific differences in the distribution of FPs in three coral taxa (Stylophora sp., Acropora sp., Echinopora sp.), combined with their photoacclimation response and that of associated symbiotic algae to light gradients in a controlled aquarium experiment. CLSM produced high-resolution images that enabled the identification of different FPs, their tissue distribution and quantification of their fluorescence intensity, as well as quantification of symbiont chlorophyll a (chl-a) fluorescence. Emission scans revealed three emission peaks between 490 - 501 nm (cyan, CFPs), 510 - 515 nm (green, GFPs), and 679 nm (chl-a fluorescence signal; Fchl) shared by all three studied species. The distribution of GFPs in Stylophora was concentrated in the intermesenterial muscle bands of the polyp, whereas CFPs were typically located at the tips of the tentacles. In contrast, Acropora and Echinopora exhibited agglomeration of CFPs and GFPs primarily in the epidermis. In general, species-specific differences in FP distribution remained unaltered during the experiment. However, linear regression models showed a significant negative relationship between CFP fluorescence intensity and light irradiance in Stylophora, whereas Echinopora exhibited a negative relation between chlorophyll fluorescence (Fchl) and light. In summary, the CLSM methodology provided a high-resolution tool to study coral FP patterns and symbiont response to irradiance, revealing ecophysiological differences among coral species at the tissue and cellular levels. CLSM has the potential to elucidate the intricacies of coral photobiology within the natural environment and to discern their adaptive responses in situ.
- Coral garden conservation and restoration: how host taxon and ex-situ maintenance affect the microbiome of soft and hard coralsPublication . Rola, Marcellina; Coelho, Márcio A. G.; Pruckner, Christian; Quiroga-Pérez, Manuela; Stock, Willem; Baylina, Núria; Engelen, Aschwin; Wägele, Heike; Serrao, Ester A.; Frade, Pedro R.Temperate coral gardens are dense coral formations, which support rich marine species diversity, enabling benthic-pelagic coupling. Over the past decades, coral gardens have been increasingly threatened by bottom fishing, oil and gas exploitation, and climate change. Microbiome research bears great potential for assisted resilience in targeted conservation and restoration approaches. Yet, fundamental parameters of the coral garden microbiome remain poorly understood. Here, we provide a first broad record of bacterial communities associated with NE Atlantic coral garden corals and their community changes as response to human maintenance in conservation research. Octocorals (10 species), scleractinians (2 species) and one black coral species, were opportunistically collected from fisheries bycatch at 60-480 m depth around Cape St. Vincent (SW Portugal). Metabarcoding of the 16S-rRNA gene using third-generation sequencing revealed a high microbial host-specificity in the wild-collected coral species analyzed, and supported the importance of bacterial families Endozoicomonadaceae (mean relative abundance +/- SE; 28.3 +/- 10.5%), Spirochaetaceae (8.2 +/- 5.8%) and Spongiibacteraceae (4.6 +/- 1.8%). Endozoicomonadaceae were particularly dominant in the octocoral order Malacalcyonacea (67.7 +/- 14.5%). The low microbial alpha diversity and limited interspecies differences among the Malacalcyonacea species suggest a conserved microbiome within this group, as compared to orders Scleralcyonacea, Antipatharia, and Scleractinia. Microbial responses to ex-situ maintenance of two branching octocoral species, Eunicella verrucosa and Paramuricea cf. grayi (Order Malacalcyonacea), were investigated (1) over 45 days under standardized aquaria conditions in the research station (Ramalhete Marine Station, CCMAR) and (2) over long-term captivity in two public aquaria, Ocean & aacute;rio de Lisboa and Zoomarine. Eunicella verrucosa displayed a stronger microbial community shift to short-term captivity (45 days), in contrast to greater microbiome stability in P. cf. grayi. However, long-term captivity in public aquaria led to microbiome shifts in both species. The strong host specificity of microbial diversity and its response to maintenance indicate that conservation and restoration of coral gardens require taxon-specific strategies.
- The microbiome of coral surface mucus has a key role in mediating holobiont health and survival upon disturbancePublication . Glasl, Bettina; Herndl, Gerhard J.; Frade, Pedro R.Microbes are well-recognized members of the coral holobiont. However, little is known about the short-term dynamics of mucus-associated microbial communities under natural conditions and after disturbances, and how these dynamics relate to the host's health. Here we examined the natural variability of prokaryotic communities (based on 16S ribosomal RNA gene amplicon sequencing) associating with the surface mucus layer (SML) of Porites astreoides, a species exhibiting cyclical mucus aging and shedding. Shifts in the prokaryotic community composition during mucus aging led to the prevalence of opportunistic and potentially pathogenic bacteria (Verrucomicrobiaceae and Vibrionaceae) in aged mucus and to a twofold increase in prokaryotic abundance. After the release of aged mucus sheets, the community reverted to its original state, dominated by Endozoicimonaceae and Oxalobacteraceae. Furthermore, we followed the fate of the coral holobiont upon depletion of its natural mucus microbiome through antibiotics treatment. After re-introduction to the reef, healthy-looking microbe-depleted corals started exhibiting clear signs of bleaching and necrosis. Recovery versus mortality of the P. astreoides holobiont was related to the degree of change in abundance distribution of the mucus microbiome. We conclude that the natural prokaryotic community inhabiting the coral SML contributes to coral health and that cyclical mucus shedding has a key role in coral microbiome dynamics.