Angelina Lo Giudice

Bioactive volatile organic compounds from Antarctic (sponges) bacteria.

Antarctic bacteria represent a reservoir of unexplored biodiversity, which, in turn, might be correlated to the synthesis of still undescribed bioactive molecules, such as antibiotics. In this work we have further characterized a panel of four marine Antarctic bacteria able to inhibit the growth of human opportunistic multiresistant pathogenic bacteria belonging to the Burkholderia cepacia complex (responsible for the cepacia syndrome in Cystic Fibrosis patients) through the production of a set of microbial Volatile Organic Compounds (mVOCs). A list of 30 different mVOCs synthesized under aerobic conditions by Antarctic bacteria was identified by GC-SPME analysis. Cross-streaking experiments suggested that Antarctic bacteria might also synthesize non-volatile molecules able to enhance the anti-Burkholderia activity. The biosynthesis of such a mixture of mVOCs was very probably influenced by both the presence/absence of oxygen and the composition of media used to grow the Antarctic strains. The antimicrobial activity exhibited by Antarctic strains also appeared to be more related to their taxonomical position rather than to the sampling site. Different Bcc bacteria were differently sensitive to the Antarctic mVOCs and this was apparently related neither to the taxonomical position of the different strains nor to their source. The genome sequence of three new Antarctic strains was determined revealing that only P. atlantica TB41 possesses some genes belonging to the nrps-pks cluster. The comparative genomic analysis performed on the genome of the four strains also revealed the presence of a few genes belonging to the core genome and involved in the secondary metabolites biosynthesis. Data obtained suggest that the antimicrobial activity exhibited by Antarctic bacteria might rely on a (complex) mixture of mVOCs whose relative concentration may vary depending on the growth conditions. Besides, it is also possible that the biosynthesis of these compounds might occur through still unknown metabolic pathways.

Biosurfactant production by Arctic and Antarctic bacteria growing on hydrocarbons

AbstractnThe risk of hydrocarbon contamination in marine polar areas is constantly increasing. Autochthonous bacteria, due to their ability to cope and survive under extreme environmental conditions, can play a fundamental role in the hydrocarbon degradation. The degradation process is often enhanced by the production of biosurfactant molecules. The present study reports for the first time on the isolation of biosurfactant-producing bacteria from Arctic and Antarctic shoreline sediments. A total of 199 psychrotolerant bacterial isolates were obtained from hydrocarbon-amended (with crude or diesel oil) microcosms. A total of 18 isolates were selected for their ability to grow in the presence of crude oil and produce biosurfactants, as it was revealed by the production of good E24 values (≥50xa0%) and/or reduction in the surface tension (under 30xa0mN/m). The positive response of the isolates to both tests suggests a possible production of biosurfactants with emulsifying and interfacial activities. Biosurfactant-producing isolates were mainly affiliated to the genera Rhodococcus (14 isolates), followed by Pseudomonas (two isolates), Pseudoalteromonas (one isolate) and Idiomarina (one isolate). Thin-layer chromatography of biosurfactant crude extracts revealed that the majority of the selected isolates were able to produce glycolipidic surfactants. Our results enlarge the knowledge, which is still poor and fragmentary, on biosurfactant producers from polar areas and indicate marine polar sediments as a source of bacteria with potential applications in the remediation of hydrocarbon-contaminated cold environments.n

The pangenome of (Antarctic) Pseudoalteromonas bacteria: evolutionary and functional insights

BackgroundPseudoalteromonas is a genus of ubiquitous marine bacteria used as model organisms to study the biological mechanisms involved in the adaptation to cold conditions. A remarkable feature shared by these bacteria is their ability to produce secondary metabolites with a strong antimicrobial and antitumor activity. Despite their biotechnological relevance, representatives of this genus are still lacking (with few exceptions) an extensive genomic characterization, including features involved in the evolution of secondary metabolites production. Indeed, biotechnological applications would greatly benefit from such analysis.ResultsHere, we analyzed the genomes of 38 strains belonging to different Pseudoalteromonas species and isolated from diverse ecological niches, including extreme ones (i.e. Antarctica). These sequences were used to reconstruct the largest Pseudoalteromonas pangenome computed so far, including also the two main groups of Pseudoalteromonas strains (pigmented and not pigmented strains). The downstream analyses were conducted to describe the genomic diversity, both at genus and group levels. This allowed highlighting a remarkable genomic heterogeneity, even for closely related strains. We drafted all the main evolutionary steps that led to the current structure and gene content of Pseudoalteromonas representatives. These, most likely, included an extensive genome reduction and a strong contribution of Horizontal Gene Transfer (HGT), which affected biotechnologically relevant gene sets and occurred in a strain-specific fashion. Furthermore, this study also identified the genomic determinants related to some of the most interesting features of the Pseudoalteromonas representatives, such as the production of secondary metabolites, the adaptation to cold temperatures and the resistance to abiotic compounds.ConclusionsThis study poses the bases for a comprehensive understanding of the evolutionary trajectories followed in time by this peculiar bacterial genus and for a focused exploitation of their biotechnological potential.

Prokaryotic Community in Lacustrine Sediments of Byers Peninsula (Livingston Island, Maritime Antarctica)

Byers Peninsula (Livingston Island, Antarctica), the largest seasonally ice-free region of the Maritime Antarctica, holds a large number of lakes, ponds, and streams. The prokaryotic structure and bacterial diversity in sediment samples collected during the 2008–2009 austral summer from five inland lakes, two coastal lakes, and an estuarine site were analyzed by Catalyzed Reporter Deposition Fluorescence In Situ Hybridization (CARD-FISH) and 16S rRNA 454 tag pyrosequencing techniques, respectively. Differently from inland lakes, which range around the oligotrophic status, coastal lakes are eutrophic environments, enriched by nutrient inputs from marine animals. Although the prokaryotic abundances (estimated as DAPI stained cells) in sediment samples were quite similar among inland and coastal lakes, Bacteria always far dominated over Archaea. Despite the phylogenetic analysis indicated that most of sequences were affiliated to a few taxonomic groups, mainly referred to Proteobacteria, Bacteroidetes, and Actinobacteria, their relative abundances greatly differed from each site. Differences in bacterial composition showed that lacustrine sediments were more phyla rich than the estuarine sediment. Proteobacterial classes in lacustrine samples were dominated by Betaproteobacteria (followed by Alphaproteobacteria, Deltaproteobacteria, and Gammaproteobacteria), while in the estuarine sample, they were mainly related to Gammaproteobacteria (followed by Deltaproteobacteria, Epsilonproteobacteria, Alphaproteobacteria, and Betaproteobacteria). Higher number of sequences of Alphaproteobacteria, Cyanobacteria, Verrucomicrobia, and Planctomycetes were observed in sediments of inland lakes compared to those of coastal lakes, whereas Chloroflexi were relatively more abundant in the sediments of coastal eutrophic lakes. As demonstrated by the great number of dominant bacterial genera, bacterial diversity was higher in the sediments of inland lakes than that in coastal lakes. Ilumatobacter (Actinobacteria), Gp16 (Acidobacteria), and Gemmatimonas (Gemmatimonadetes) were recovered as dominant genera in both inland and coastal lakes, but not in the estuarine sample, indicating that they may be useful markers of Antarctic lakes. The proximity to the sea, the different lake depths and the external or internal origin of the nutrient sources shape the bacterial communities composition in lacustrine sediments of Byers Peninsula.

Cold-adapted bacteria from a coastal area of the Ross Sea (Terra Nova Bay, Antarctica): linking microbial ecology to biotechnology

AbstractnThis review covers available information concerning the bacterial communities inhabiting coastal areas of Terra Nova Bay (Antarctica). Research was mainly focused on seawater, sediment, and benthic filter feeders. Coping with adverse environmental conditions, Antarctic bacteria have evolved peculiar strategies to survive in this harsh environment. In addition to cellular modifications, antagonistic interactions between them might have contributed to their adaptation to permanently low temperatures by reducing the presence of microbial competitors. The interrelationships existing among the members of bacterial communities, between them and their surrounding environment will be discussed. Quite interestingly, environmental selective pressures have led to the evolution of metabolic pathways responsible for the synthesis of unique secondary metabolites exhibiting a variety of biological activities. Finally, human perturbations have posed a serious questioning about the ability of microbial communities to respond to environmental anthropogenic changes in Antarctica. From a biotechnological viewpoint, bacteria inhabiting Terra Nova Bay are intriguing; indeed, many of them are able to degrade hydrocarbons and polychlorinated biphenyls at low temperatures, and tolerate heavy metals and antibiotics. After a brief description of cold-adapted bacteria, we will report on ecological roles of bacterial communities inhabiting the Terra Nova Bay, alongside a focus on biotechnological aspects of their cultivable fraction.

Antimicrobial activity of Pseudoalteromonas strains isolated from the Ross Sea (Antarctica) versus Cystic Fibrosis opportunistic pathogens

AbstractnIn Antarctica, the selective pressure may have led to the evolution of novel capabilities by indigenous organisms, including microorganisms, to achieve competitive advantages. In this work, the ability of thirteen Antarctic Pseudoalteromonas isolates from different sources (sponges, seawater and sediments) to synthesize antimicrobial compounds was analysed. The antibacterial activity was tested against Cystic Fibrosis opportunistic pathogens belonging to the Burkholderia cepacia complex (Bcc). Data obtained revealed that all the Pseudoalteromonas strains synthesize a plethora of microbial volatile organic compounds (mVOCs) and diffusible molecules that strongly interfere with the growth of Bcc bacteria and that this synthesis may be influenced by the growth media essentially in terms of amount of each mVOC. The finding that mVOCs profiles can be obtained from bacteria belonging to very different taxa strongly suggests that the synthesis of such compounds might have a great relevance from an evolutionary and/or ecological viewpoint. Since these mVOCs are able to completely inhibit the growth of Bcc bacteria, thus exhibiting an antibacterial activity, it is possible that such compounds might represent one of the forces driving the structuring of bacterial communities inhabiting the same ecological niche.

Ecology of cold environments: new insights of bacterial metabolic adaptation through an integrated genomic-phenomic approach

Cold environments dominate Earth’s biosphere, hosting complex microbial communities with the ability to thrive at low temperatures. However, the underlying molecular mechanisms and the metabolic pathways involved in bacterial cold-adaptation mechanisms are still not fully understood. Herein, we assessed the metabolic features of the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 (PhTAC125), a model organism for cold-adaptation, at both 4u2009°C and 15u2009°C, by integrating genomic and phenomic (high-throughput phenotyping) data and comparing the obtained results to the taxonomically related Antarctic bacterium Pseudoalteromonas sp. TB41 (PspTB41). Although the genome size of PspTB41 is considerably larger than PhTAC125, the higher number of genes did not reflect any higher metabolic versatility at 4u2009°C as compared to PhTAC125. Remarkably, protein S-thiolation regulated by glutathione and glutathionylspermidine appeared to be a new possible mechanism for cold adaptation in PhTAC125. More in general, this study represents an example of how ‘multi-omic’ information might potentially contribute in filling the gap between genotypic and phenotypic features related to cold-adaptation mechanisms in bacteria.

Enrichment, isolation and biodegradation potential of psychrotolerant polychlorinated-biphenyl degrading bacteria from the Kongsfjorden (Svalbard Islands, High Arctic Norway)☆

Persistent organic pollutants (POPs), such as polychlorinated biphenyls (PCBs), have been detected in abiotic Arctic matrices: surface sediments and seawater from coastal areas in the Kongsfjorden were collected and analyzed. Levels of PCBs varied depending on the sampling site. Total PCB concentrations were between 11.63 (site C2W) and 27.69pgl-1 (site AW). These levels were comparable to those reported previously in lake sediments from the northern Svalbard. The occurrence and biodegradation potential of cold-adapted PCB-oxidizing bacteria in seawater and sediment along the fjord was also evaluated. After enrichment with biphenyl, 246 isolates were obtained with 45 of them that were able to grow in the presence of the PCB mixture Aroclor 1242, as the sole carbon source. The catabolic gene bphA was harbored by 17 isolates with affiliates to the genera Algoriphagus, Devosia and Salinibacterium that have been never reported as able to utilize PCBs, thus deserving further investigation. The total removal of Aroclor 1242 and selected PCB congeners was evaluated at 4 and 15°C for eight bphA-harboring isolates and Gelidibacter sp. DS-10. With few exceptions, tested strains showed greater efficiency at 15 than at 4°C. Isolates were able to reduce most chromatographic peaks by >50%, with some di- and trichlorobiphenyls that were quite totally removed (>90%).

Genome-scale phylogenetic and DNA composition analyses of Antarctic Pseudoalteromonas bacteria reveal inconsistencies in current taxonomic affiliation

Bacteria belonging to the Pseudoalteromonas genus have important ecological implications in marine environments, playing a role in the control of microbial community as producers of bioactive molecules endowed with antifouling activity and able to antagonize larvae, fungi and bacteria, including important human pathogens. For these reasons, representatives of this genus are very promising for biotechnological and biomedical applications. In this work, we used different genome-scale approaches to infer the taxonomy of 38 Pseudoalteromonas representatives (most of which isolated from Antarctica) and whose complete genome has been sequenced. We show that an accurate re-evaluation of the real taxonomic relationships of Pseudoalteromonas representatives is needed since many inconsistencies with the current taxonomic annotation were observed. Moreover, data obtained with different genome-scale methods are consistent, confirming the reliability of the genomic approaches. On the basis of these data, we propose a re-annotation for some Pseudoalteromonas species. This proposal should be validated in the future by comparing the phenotypes of these strains.

Extracellular polymeric substances with metal adsorption capacity produced by Pseudoalteromonas sp. MER144 from Antarctic seawater

The EPS-producing Pseudoalteromonas sp. MER144 was selected among 606 isolates from Antarctic seawater due to its evident slimy appearance on agar plates. The production of EPSs was enhanced by a step-by-step approach varying the carbon source, substrate and NaCl concentrations, temperature, and pH. Optimal conditions for the EPS production resulted at temperature of 4xa0°C and pHxa07, with addition of 2% sucrose (w/v) and 3% NaCl (w/v). EPSs produced under optimal conditions were chemically characterized, resulting in a moderate carbohydrate content (35%), uronic acids (14%), and proteins (12%). Monosaccharide composition was estimated to be Glu:Man:GluN:Ara:GluA:GalA:Gal (1:0.36:0.26:0.06:0.06:0.05:0.03), while the estimated molecular weight was about 250xa0kDa. The addition of sucrose in the culture medium, by stimulating the EPS production, allowed MER144 to tolerate higher concentrations of mercury and cadmium. This finding was probably dependent on the presence of uronic acids and sulfate groups, which can bind cations, in the extracted EPSs. Monitoring EPS production under optimal conditions at different concentrations of mercury and cadmium revealed that EPS amounts increased at increasing heavy metal concentrations, indicating an adaptation to the stress conditions tested.