Isabella Gandolfi

Microbial biosurfactants production, applications and future potential

Microorganisms synthesise a wide range of surface-active compounds (SAC), generally called biosurfactants. These compounds are mainly classified according to their molecular weight, physico-chemical properties and mode of action. The low-molecular-weight SACs or biosurfactants reduce the surface tension at the air/water interfaces and the interfacial tension at oil/water interfaces, whereas the high-molecular-weight SACs, also called bioemulsifiers, are more effective in stabilising oil-in-water emulsions. Biosurfactants are attracting much interest due to their potential advantages over their synthetic counterparts in many fields spanning environmental, food, biomedical, and other industrial applications. Their large-scale application and production, however, are currently limited by the high cost of production and by limited understanding of their interactions with cells and with the abiotic environment. In this paper, we review the current knowledge and the latest advances in biosurfactant applications and the biotechnological strategies being developed for improving production processes and future potential.

Influence of compost amendment on microbial community and ecotoxicity of hydrocarbon-contaminated soils.

The influence of a high quality compost amendment on two soils contaminated with diesel oil and polycyclic aromatic hydrocarbons, PAHs, respectively, was evaluated, with respect to contaminant biodegradation, microbial community composition and soil toxicity and genotoxicity. For each of the two soils, two 20-kg biopiles were set up, one without amendments and one compost-amended. GC/FID analyses revealed that compost was effective in enhancing biodegradation of diesel oil and of four-ring PAHs. It also influenced microbial community composition, as inferred by ARDRA analyses and partial sequencing of 16S rDNA of clones from libraries constructed from each soil sample. Microtox analyses on soil aqueous saline extracts and Solid Phase Tests showed some toxicity reduction due to compost addition, while the Comet assay, performed on coelomocytes of earthworms exposed to contaminated soils, did not show genotoxicity reduction. In general, the use of compost amendment to hydrocarbon-contaminated soils in a bioremediation process proved to be effective for depletion of contaminants and reduction of toxicity.

Temporal variability and effect of environmental variables on airborne bacterial communities in an urban area of Northern Italy

Despite airborne microorganisms representing a relevant fraction of atmospheric suspended particles, only a small amount of information is currently available on their abundance and diversity and very few studies have investigated the environmental factors influencing the structure of airborne bacterial communities. In this work, we used quantitative PCR and Illumina technology to provide a thorough description of airborne bacterial communities in the urban area of Milan (Italy). Forty samples were collected in 10-day sampling sessions, with one session per season. The mean bacterial abundance was about 104 ribosomal operons per m3 of air and was lower in winter than in the other seasons. Communities were dominated by Actinobacteridae, Clostridiales, Sphingobacteriales and few proteobacterial orders (Burkholderiales, Rhizobiales, Sphingomonadales and Pseudomonadales). Chloroplasts were abundant in all samples. A higher abundance of Actinobacteridae, which are typical soil-inhabiting bacteria, and a lower abundance of chloroplasts in samples collected on cold days were observed. The variation in community composition observed within seasons was comparable to that observed between seasons, thus suggesting that airborne bacterial communities show large temporal variability, even between consecutive days. The structure of airborne bacterial communities therefore suggests that soil and plants are the sources which contribute most to the airborne communities of Milan atmosphere, but the structure of the bacterial community seems to depend mainly on the source of bacteria that predominates in a given period of time.

Unravelling the bacterial diversity in the atmosphere.

The study of airborne biological particles (‘bioaerosols’) has gained interest in recent years, due to an increasing amount of evidence suggesting that this fraction of airborne particulate matter may play a critical role in the negative effects of aerosols on biological systems. Pioneer investigations demonstrated that bacteria do exist in the atmosphere and can be metabolically active, although studies have not proved whether they actually form ecological communities or are merely assemblages of organisms passively transported from different sources. For a long time, cultivation-based methods have been the gold standard to describe and quantify airborne microorganisms. However, the use of culture-independent techniques and, more recently, of the next-generation sequencing-based methods, has improved the ability of the scientific community to investigate bioaerosols in detail and to address further research questions, such as the temporal and spatial variability of airborne bacterial assemblages, the environmental factors affecting this variability and the potential sources of atmospheric bacteria. This paper provides a systematic review of the state-of-the-art methodologies used in the study of airborne bacteria to achieve each of the aforementioned research objectives, as well as the main results obtained so far. Critical evaluations of the current state of the knowledge and suggestions for further researches are provided.

Bacterial community structure on two alpine debris-covered glaciers and biogeography of Polaromonas phylotypes.

High-elevation cold environments are considered ideal places to test hypotheses about mechanisms of bacterial colonization and succession, and about bacterial biogeography. Debris-covered glaciers (glaciers whose ablation area is mainly covered by a continuous layer of rock debris fallen from the surrounding mountains) have never been investigated in this respect so far. We used the Illumina technology to analyse the V5 and V6 hypervariable regions of the bacterial 16S rRNA gene amplified from 38 samples collected in July and September 2009 at different distances from the terminus on two debris-covered glaciers (Miage and Belvedere—Italian Alps). Heterotrophic taxa-dominated communities and bacterial community structure changed according to ice ablation rate, organic carbon content of the debris and distance from the glacier terminus. Bacterial communities therefore change during downwards debris transport, and organic carbon of these recently exposed substrates is probably provided more by allochthonous deposition of organic matter than by primary production by autotrophic organisms. We also investigated whether phylotypes of the genus Polaromonas, which is ubiquitous in cold environments, do present a biogeographical distribution by analysing the sequences retrieved in this study together with others available in the literature. We found that the genetic distance among phylotypes increased with geographic distance; however, more focused analyses using discrete distance classes revealed that both sequences collected at sites <100 km and at sites 9400–13 500 km to each other were more similar than those collected at other distance classes. Evidences of biogeographic distribution of Polaromonas phylotypes were therefore contrasting.

Environmental fate, toxicity, characteristics and potential applications of novel bioemulsifiers produced by Variovorax paradoxus 7bCT5

The aims of this work were the characterisation and the evaluation of potential environmental applications of the bioemulsifiers produced by Variovorax paradoxus 7bCT5. V. paradoxus 7bCT5 produces a mixture of high molecular weight polysaccharides. The extracellular bioemulsifiers were able to produce a thick stable oil/water emulsion and maintained the emulsification activity after boiling and at low temperatures. Environmental behavior and impact of bioemulsifiers release were assessed by evaluating biodegradability, toxicity and soil sorption. Respirometric tests showed that moderate biodegradability occurred by soil bacterial inoculum. Furthermore, the produced compounds did not show any toxic properties through different ecotoxicological tests. The K(d) values ranged from 1.3 to 7.3 L/kg indicating a high sorption affinity of the bioemulsifier molecules to soil particles. The soil sorption affinity likely affected the bioemulsifier ability to remove hydrocarbons from contaminated soils. In fact, V. paradoxus 7bCT5 bioemulsifiers significantly increased the removal of crude-oil from sandy soil compared to water.

Anodic and cathodic microbial communities in single chamber microbial fuel cells

Microbial fuel cells (MFCs) are a rapidly growing technology for energy production from wastewater and biomasses. In a MFC, a microbial biofilm oxidizes organic matter and transfers electrons from reduced compounds to an anode as the electron acceptor by extracellular electron transfer (EET). The aim of this work was to characterize the microbial communities operating in a Single Chamber Microbial Fuel Cell (SCMFC) fed with acetate and inoculated with a biogas digestate in order to gain more insight into anodic and cathodic EET. Taxonomic characterization of the communities was carried out by Illumina sequencing of a fragment of the 16S rRNA gene. Microorganisms belonging to Geovibrio genus and purple non-sulfur (PNS) bacteria were found to be dominant in the anodic biofilm. The alkaliphilic genus Nitrincola and anaerobic microorganisms belonging to Porphyromonadaceae family were the most abundant bacteria in the cathodic biofilm.

Light-dependent microbial metabolisms drive carbon fluxes on glacier surfaces.

Biological processes on glacier surfaces affect glacier reflectance, influence surface energy budget and glacier response to climate warming, and determine glacier carbon exchange with the atmosphere. Currently, carbon balance of supraglacial environment is assessed as the balance between the activity of oxygenic phototrophs and the respiration rate of heterotrophic organisms. Here we present a metagenomic analysis of tiny wind-blown supraglacial sediment (cryoconite) from Baltoro (Pakistani Karakoram) and Forni (Italian Alps) glaciers, providing evidence for the occurrence in these environments of different and previously neglected metabolic pathways. Indeed, we observed high abundance of heterotrophic anoxygenic phototrophs, suggesting that light might directly supplement the energy demand of some bacterial strains allowing them to use as carbon source organic molecules, which otherwise would be respired. Furthermore, data suggest that CO2 could be produced also by microbiologically mediated oxidation of CO, which may be produced by photodegradation of organic matter.

Hydrocarbon degrading microbial communities in bench scale aerobic biobarriers for gasoline contaminated groundwater treatment

BTEX compounds (benzene, toluene, ethylbenzene and xylenes) and methyl tert-butyl ether (MTBE) are some of the main constituents of gasoline and can be accidentally released in the environment. In this work the effect of bioaugmentation on the microbial communities in a bench scale aerobic biobarrier for gasoline contaminated water treatment was studied by 16S rRNA gene sequencing. Catabolic genes (tmoA and xylM) were quantified by qPCR, in order to estimate the biodegradation potential, and the abundance of total bacteria was estimated by the quantification of the number of copies of the 16S rRNA gene. Hydrocarbon concentration was monitored over time and no difference in the removal efficiency for the tested conditions was observed, either with or without the microbial inoculum. In the column without the inoculum the most abundant genera were Acidovorax, Bdellovibrio, Hydrogenophaga, Pseudoxanthomonas and Serpens at the beginning of the column, while at the end of the column Thauera became dominant. In the inoculated test the microbial inoculum, composed by Rhodococcus sp. CE461, Rhodococcus sp. CT451 and Methylibium petroleiphilum LMG 22953, was outcompeted. Quantitative PCR results showed an increasing in xylM copy number, indicating that hydrocarbon degrading bacteria were selected during the treatment, although only a low increase of the total biomass was observed. However, the bioaugmentation did not lead to an increase in the degradative potential of the microbial communities.

Antibiotic resistance in bacteria associated with coarse atmospheric particulate matter in an urban area

Aims:  To assess antibiotic resistance in airborne bacteria associated with coarse particulate matter (PM10) in an urban area, with specific considerations about the Staphylococcus genus.