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Browsing ESS - SA - Artigos by Author "Alexandrino, Diogo A. M."
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- Actinobacteria from arctic and atlantic deep-sea sediments—biodiversity and bioactive potentialPublication . Ribeiro, Inês ; Antunes, Jorge T. ; Alexandrino, Diogo A. M.; Tomasino, Maria Paola ; Almeida, Eduarda ; Hilário, Ana ; Urbatzka, Ralph ; Leão, Pedro N. ; Mucha, Ana P. ; Carvalho, Maria F.The deep-sea covers over 70% of the Earth’s surface and harbors predominantly uncharacterized bacterial communities. Actinobacteria are the major prokaryotic source of bioactive natural products that find their way into drug discovery programs, and the deep-sea is a promising source of biotechnologically relevant actinobacteria. Previous studies on actinobacteria in deep-sea sediments were either regionally restricted or did not combine a community characterization with the analysis of their bioactive potential. Here we characterized the actinobacterial communities of upper layers of deep-sea sediments from the Arctic and the Atlantic (Azores and Madeira) ocean basins, employing 16S rRNA metabarcoding, and studied the biosynthetic potential of cultivable actinobacteria retrieved from those samples. Metabarcoding analysis showed that the actinobacterial composition varied between the sampled regions, with higher abundance in the Arctic samples but higher diversity in the Atlantic ones. Twenty actinobacterial genera were detected using metabarcoding, as a culture-independent method, while culture-dependent methods only allowed the identification of nine genera. Isolation of actinobacteria resulted on the retrieval of 44 isolates, mainly associated with Brachybacterium, Microbacterium, and Brevibacterium genera. Some of these isolates were only identified on a specific sampled region. Chemical extracts of the actinobacterial isolates were subsequently screened for their antimicrobial, anticancer and anti-inflammatory activities. Extracts from two Streptomyces strains demonstrated activity against Candida albicans. Additionally, eight extracts (obtained from Brachybacterium, Brevibacterium, Microbacterium, Rhodococcus, and Streptomyces isolates) showed significant activity against at least one of the tested cancer cell lines (HepG2 and T-47D). Furthermore, 15 actinobacterial extracts showed anti-inflammatory potential in the RAW 264.4 cell model assay, with no concomitant cytotoxic response. Dereplication and molecular networking analysis of the bioactive actinobacterial extracts showed the presence of some metabolites associated with known natural products, but one of the analyzed clusters did not show any match with the natural products described as responsible for these bioactivities. Overall, we were able to recover taxonomically diverse actinobacteria with different bioactivities from the studied deep-sea samples. The conjugation of culture-dependent and -independent methods allows a better understanding of the actinobacterial diversity of deep-sea environments, which is important for the optimization of approaches to obtain novel chemically-rich isolates.
- Combined effects of temperature and dietary lipid level on body composition, growth, and freshness profile in european seabass, dicentrarchus labraxPublication . Cardoso, Patrícia G.; Gonçalves, Odete; Cavalheri, Thais; Amorim, Vânia E.; Cao, Weiwei; Alexandrino, Diogo A. M.; Jia, Zhongjun; Carvalho, Maria F.; Vaz-Pires, Paulo; Ozório, Rodrigo O. A.The effects of increasing temperature and dietary lipid level on the body composition, growth performance, and freshness profile of the European seabass (Dicentrarchus labrax) were evaluated through a fish trial lasting 56 days. Findings demonstrated that fish reared at 24 °C presented a lower lipid level and a higher daily growth index than those reared at 20 °C. On the other hand, the sea bass condition index did not change among treatments. Additionally, sensory analysis (the Quality Index Method) and microbiological analysis revealed that fish reared at 24 °C showed better freshness conditions than those at 20 °C. Nevertheless, the dietary lipid level did not have any influence on fish freshness conditions. Therefore, our data suggest that the increase in temperature to 24 °C is beneficial for the growth and freshness profile of this particular species in aquaculture.
- Complementary field and laboratory batch studies to quantify generation rates of perfluoroalkyl acids in a contaminated agricultural topsoil with unknown precursorsPublication . Haluska, Alexander Arthur; Röhler, Klaus; Fabregat-Palau, Joel; Alexandrino, Diogo A. M.; Abramov, Sergey; Thompson, Katharine J.; Straub, Daniel; Kleindienst, Sara; Bugsel, Boris; Zweigle, Jonathan; Zwiener, Christian; Grathwohl, Peter; Alexandrino, DiogoSoil microbiome changes and generation rates of per- and polyfluoroalkyl substances (PFAS) precursors were studied in a contaminated agricultural field using a combination of field and laboratory batch microcosm studies. 16S rRNA gene amplicon sequencing was used to track how microbial community composition changed over time, while perfluoroalkyl acids (PFAA) generation rates were quantified using a combination of field and batch incubations combined with the direct total oxidizable precursor (dTOP) assay. The study site in Brilon-Scharfenberg, North Rhine-Westphalia, Germany, has PFAS contamination in the topsoil (0 to 30cm) originating from compost. Generation rate constants of these short-chain PFAA estimated from batch incubations (0.12 to 0.751/year) were higher but similar to rate constants from the fields (0.05 to 0.221/year). Long-term field mass discharge data (2009 to 2023) suggest that at least 60years are needed to remove 99.99% of short-chain PFAA and their precursors. 16S rRNA gene amplicon sequencing data revealed a major impact of PFAA on the biodiversity of soil microorganisms, with batch-incubated contaminated soils showing higher richness and diversity indexes than field control soils. However, most of these impacts occurred at lower taxonomical ranks and did not seem to have a prominent impact on the overall structure of the autochtonous microbial communities of the soils where PFAA were produced and accumulated. Overall, our findings demonstrate that well-controlled aerobic batch test combined with the results of dTOP assay are a suitable approach for estimating short-chain PFAA generation rates. Additionally, our research suggests that the complete removal of PFAA precursors from topsoil will take decades.
- Unveiling the culturable and non‐culturable actinobacterial diversity in two macroalgae species from the northern Portuguese coastPublication . Girão, Mariana; Alexandrino, Diogo A. M.; Cao, Weiwei; Costa, Isabel; Jia, Zhongjun; Carvalho, Maria F.; Alexandrino, DiogoActinomycetota, associated with macroalgae, remains one of the least explored marine niches. The secondary metabolism of Actinomycetota, the primary microbial source of compounds relevant to biotechnology, continues to drive research into the distribution, dynamics, and metabolome of these microorganisms. In this study, we employed a combination of traditional cultivation and metagenomic analysis to investigate the diversity of Actinomycetota in two native macroalgae species from the Portuguese coast. We obtained and taxonomically identified a collection of 380 strains, which were distributed across 12 orders, 15 families, and 25 genera affiliated with the Actinomycetia class, with Streptomyces making up approximately 60% of the composition. Metagenomic results revealed the presence of Actinomycetota in both Chondrus crispus and Codium tomentosum datasets, with relative abundances of 11% and 2%, respectively. This approach identified 12 orders, 16 families, and 17 genera affiliated with Actinomycetota, with minimal overlap with the cultivation results. Acidimicrobiales emerged as the dominant actinobacterial order in both macroalgae, although no strain affiliated with this taxonomic group was successfully isolated. Our findings suggest that macroalgae represent a hotspot for Actinomycetota. The synergistic use of both culture-dependent and independent approaches proved beneficial, enabling the identification and recovery of not only abundant but also rare taxonomic members.