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- Multitaxon activity profiling reveals differential microbial response to reduced seawater pH and oil pollutionPublication . Coelho, Francisco J. R. C.; Cleary, Daniel F. R.; Costa, Rodrigo; Ferreira, Marina; Polonia, Ana R. M.; Silva, Artur M. S.; Simoes, Mario M. Q.; Oliveira, Vanessa; Gomes, Newton C. M.There is growing concern that predicted changes to global ocean chemistry will interact with anthropogenic pollution to significantly alter marine microbial composition and function. However, knowledge of the compounding effects of climate change stressors and anthropogenic pollution is limited. Here, we used 16S and 18S rRNA (cDNA)-based activity profiling to investigate the differential responses of selected microbial taxa to ocean acidification and oil hydrocarbon contamination under controlled laboratory conditions. Our results revealed that a lower relative abundance of sulphate-reducing bacteria (Desulfosarcina/Desulfococcus clade) due to an adverse effect of seawater acidification and oil hydrocarbon contamination (reduced pH-oil treatment) may be coupled to changes in sediment archaeal communities. In particular, we observed a pronounced compositional shift and marked reduction in the prevalence of otherwise abundant operational taxonomic units (OTUs) belonging to the archaeal Marine Benthic Group B and Marine Hydrothermal Vent Group (MHVG) in the reduced pH-oil treatment. Conversely, the abundance of several putative hydrocarbonoclastic fungal OTUs was higher in the reduced pH-oil treatment. Sediment hydrocarbon profiling, furthermore, revealed higher concentrations of several alkanes in the reduced pH-oil treatment, corroborating the functional implications of the structural changes to microbial community composition. Collectively, our results advance the understanding of the response of a complex microbial community to the interaction between reduced pH and anthropogenic pollution. In future acidified marine environments, oil hydrocarbon contamination may alter the typical mixotrophic and k-/r-strategist composition of surface sediment microbiomes towards a more heterotrophic state with lower doubling rates, thereby impairing the ability of the ecosystem to recover from acute oil contamination events.
- A bacterial consortium isolated from an Icelandic fumarole displays exceptionally high levels of sulfate reduction and metals resistancePublication . Alexandrino, Maria; Macías, Francisco; Da Silva Costa, Rodrigo; Gomes, Newton C. M.; Canario, Adelino V. M.; Costa, Maria ClaraThe soils of three fumaroles and one mining site, all with high metal content, were surveyed for the presence of metal-resistant sulfate-reducing bacteria and their potential application in the bioremediation of acid mine drainages. By means of selective soil enrichments a bacterial consortium was isolated from an Icelandic fumarole that displayed very high sulfate reduction in the presence of a mixture of 0.75 g/L of Fe, 0.20 g/L of Zn and 0.080 g/L of Cu. Under these conditions the bacterial consortium reduced 91% of the added 3.9 g/L of sulfate after 28 days, precipitating 100% of the Fe, 96% of the Zn and 97% of the Cu during the same time. Both total bacterial numbers and numbers of culturable sulfate-reducing bacteria remained unchanged when grown in media containing metals, suggesting low or absent inhibitory effects of the metals on the bacterial consortium. PCR-DGGE profiles of the sulfate reducing bacterial communities obtained from the Icelandic fumarole sample showed that bacterial diversity decreased significantly after metal addition: from the original 12 ribotypes only two were detected in the metaltolerant culture. Phylogenetic analysis of 16S ribosomal RNA gene sequences revealed that these two ribotypes were affiliated with the genera Clostridium and Desulfovibrio, with C. subterminale, C. pascui, C. mesophilum and C. peptidovorans and D. desulfuricans identified as their closest relatives.
- Development and validation of an experimental life support system for assessing the effects of global climate change and environmental contamination on estuarine and coastal marine benthic communitiesPublication . Coelho, Francisco J. R. C.; Rocha, Rui J. M.; Pires, Ana C. C.; Ladeiro, Bruno; Castanheira, Jose M.; Costa, Rodrigo; Almeida, Adelaide; Cunha, Angela; Lillebo, Ana Isabel; Ribeiro, Rui; Pereira, Ruth; Lopes, Isabel; Marques, Catarina; Moreira-Santos, Matilde; Calado, Ricardo; Cleary, Daniel F. R.; Gomes, Newton C. M.An experimental life support system (ELSS) was constructed to study the interactive effects of multiple stressors on coastal and estuarine benthic communities, specifically perturbations driven by global climate change and anthropogenic environmental contamination. The ELSS allows researchers to control salinity, pH, temperature, ultraviolet radiation (UVR), tidal rhythms and exposure to selected contaminants. Unlike most microcosms previously described, our system enables true independent replication (including randomization). In addition to this, it can be assembled using commercially available materials and equipment, thereby facilitating the replication of identical experimental setups in different geographical locations. Here, we validate the reproducibility and environmental quality of the system by comparing chemical and biological parameters recorded in our ELSS with those prevalent in the natural environment. Water, sediment microbial community and ragworm (the polychaete Hediste diversicolor) samples were obtained from four microcosms after 57days of operation. In general, average concentrations of dissolved inorganic nutrients (NO3-; NH4+ and PO4-3) in the water column of the ELSS experimental control units were within the range of concentrations recorded in the natural environment. While some shifts in bacterial community composition were observed between in situ and ELSS sediment samples, the relative abundance of most metabolically active bacterial taxa appeared to be stable. In addition, ELSS operation did not significantly affect survival, oxidative stress and neurological biomarkers of the model organism Hediste diversicolor. The validation data indicate that this system can be used to assess independent or interactive effects of climate change and environmental contamination on benthic communities. Researchers will be able to simulate the effects of these stressors on processes driven by microbial communities, sediment and seawater chemistry and to evaluate potential consequences to sediment toxicity using model organisms such as Hediste diversicolor.
- Mangrove microniches determine the structural and functional diversity of enriched petroleum hydrocarbon-degrading consortiaPublication . Gomes, Newton C. M.; Flocco, Cecilia G.; Costa, Rodrigo; Junca, Howard; Vilchez, Ramiro; Pieper, Dietmar H.; Kroegerrecklenfort, Ellen; Paranhos, Rodolfo; Mendonca-Hagler, Leda C. S.; Smalla, KorneliaIn this study, the combination of culture enrichments and molecular tools was used to identify bacterial guilds, plasmids and functional genes potentially important in the process of petroleum hydrocarbon (PH) decontamination in mangrove microniches (rhizospheres and bulk sediment). In addition, we aimed to recover PH-degrading consortia (PHDC) for future use in remediation strategies. The PHDC were enriched with petroleum from rhizosphere and bulk sediment samples taken from a mangrove chronically polluted with oil hydrocarbons. Southern blot hybridization (SBH) assays of PCR amplicons from environmental DNA before enrichments resulted in weak positive signals for the functional gene types targeted, suggesting that PH-degrading genotypes and plasmids were in low abundance in the rhizosphere and bulk sediments. However, after enrichment, these genes were detected and strong microniche-dependent differences in the abundance and composition of hydrocarbonoclastic bacterial populations, plasmids (IncP-1 alpha, IncP-1 beta, IncP-7 and IncP-9) and functional genes (naphthalene, extradiol and intradiol dioxygenases) were revealed by in-depth molecular analyses [PCR-denaturing gradient gel electrophoresis and hybridization (SBH and microarray)]. Our results suggest that, despite the low abundance of PH-degrading genes and plasmids in the environmental samples, the original bacterial composition of the mangrove microniches determined the structural and functional diversity of the PHDC enriched.
- Marine sponge and octocoral-associated bacteria show versatile secondary metabolite biosynthesis potential and antimicrobial activities against human pathogensPublication . Almeida, João F.; Marques, Matilde; Oliveira, Vanessa; Egas, Conceição; Mil-Homens, Dalila; Viana, Romeu; Cleary, Daniel F. R.; Huang, Yusheng M.; Fialho, Arsénio M.; Teixeira, Miguel C.; Gomes, Newton C. M.; Costa, Rodrigo; Keller-Costa, TinaMarine microbiomes are prolific sources of bioactive natural products of potential pharmaceutical value. This study inspected two culture collections comprising 919 host-associated marine bacteria belonging to 55 genera and several thus-far unclassified lineages to identify isolates with potentially rich secondary metabolism and antimicrobial activities. Seventy representative isolates had their genomes mined for secondary metabolite biosynthetic gene clusters (SM-BGCs) and were screened for antimicrobial activities against four pathogenic bacteria and five pathogenic Candida strains. In total, 466 SM-BGCs were identified, with antimicrobial peptide- and polyketide synthase-related SM-BGCs being frequently detected. Only 38 SM-BGCs had similarities greater than 70% to SM-BGCs encoding known compounds, highlighting the potential biosynthetic novelty encoded by these genomes. Cross-streak assays showed that 33 of the 70 genome-sequenced isolates were active against at least one Candida species, while 44 isolates showed activity against at least one bacterial pathogen. Taxon-specific differences in antimicrobial activity among isolates suggested distinct molecules involved in antagonism against bacterial versus Candida pathogens. The here reported culture collections and genome-sequenced isolates constitute a valuable resource of understudied marine bacteria displaying antimicrobial activities and potential for the biosynthesis of novel secondary metabolites, holding promise for a future sustainable production of marine drug leads.
- Assessing the genomic composition, putative ecological relevance and biotechnological potential of plasmids from sponge bacterial symbiontsPublication . Oliveira, Vanessa; Polónia, Ana R. M.; Cleary, Daniel F. R.; Huang, Yusheng M.; de Voogd, Nicole J.; Keller-Costa, Tina; Costa, Rodrigo; Gomes, Newton C. M.Plasmid-mediated transfer of genes can have direct consequences in several biological processes within sponge microbial communities. However, very few studies have attempted genomic and functional characterization of plasmids from marine host-associated microbial communities in general and those of sponges in particular. In the present study, we used an endogenous plasmid isolation method to obtain plasmids from bacterial symbionts of the marine sponges Stylissa carteri and Paratetilla sp. and investigated the genomic composition, putative ecological relevance and biotechnological potential of these plasmids. In total, we isolated and characterized three complete plasmids, three plasmid prophages and one incomplete plasmid. Our results highlight the importance of plasmids to transfer relevant genetic traits putatively involved in microbial symbiont adaptation and host-microbe and microbe-microbe interactions. For example, putative genes involved in bacterial response to chemical stress, competition, metabolic versatility and mediation of bacterial colonization and pathogenicity were detected. Genes coding for enzymes and toxins of biotechnological potential were also detected. Most plasmid prophage coding sequences were, however, hypothetical proteins with unknown functions. Overall, this study highlights the ecological relevance of plasmids in the marine sponge microbiome and provides evidence that plasmids of sponge bacterial symbionts may represent an untapped resource of genes of biotechnological interest.