Andrea Franzetti

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.

Potential applications of surface active compounds by Gordonia sp. strain BS29 in soil remediation technologies.

A wide range of structurally different surface active compounds (SACs) is synthesised by many prokaryotic and eukaryotic microorganisms. Due to their properties, microbial SACs have been exploited in environmental remediation techniques. From a diesel-contaminated soil, we isolated the Gordonia sp. strain BS29 which extensively grows on aliphatic hydrocarbons and produces two different types of SACs: extracellular bioemulsans and cell-bound biosurfactants. The aim of this work was to evaluate the potential applications of the strain BS29 and its SACs in the following environmental technologies: bioremediation of soils contaminated by aliphatic and aromatic hydrocarbons, and washing of soils contaminated by crude oil, polycyclic aromatic hydrocarbons (PAHs) and heavy metals. Microcosm bioremediation experiments were carried out with soils contaminated by aliphatic hydrocarbons or PAHs, while batch soil washing experiments were carried out with soils contaminated by crude oil, PAHs or heavy metals. Bioremediation results showed that the BS29 bioemulsans are able to slightly enhance the biodegradation of recalcitrant branched hydrocarbons. On the other hand, we obtained the best results in soil washing of hydrocarbons. The BS29 bioemulsans effectively remove crude oil and PAHs from soil. Particularly, crude oil removal by BS29 bioemulsans is comparable to the rhamnolipid one in the same experimental conditions showing that the BS29 bioemulsans are promising washing agents for remediation of hydrocarbon-contaminated soils.

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.

Applications of Biological Surface Active Compounds in Remediation Technologies

Many microorganisms synthesize a wide range of surface active compounds (SACs), classified according to their molecular weights, properties and localizations. The low molecular weight SACs or biosurfactants lower the surface tension at the air/water interfaces and the interfacial tension at oil/water interfaces, whereas the high molecular weight SACs, also known as bioemulsifiers, are more effective in stabilizing oil-in-water emulsions. The ability to biosynthesize SACs is, often, coupled with the ability of these microorganisms to grow on immiscible carbon sources, such as hydrocarbons. Different mechanisms are involved in the SACs interactions between microbial cells and immiscible hydrocarbons including: (i) emulsification, (ii) micellarization, (iii) adhesion-deadhesion of microorganisms to and from hydrocarbons and (iv) desorption of contaminants. These naturally occurring phenomena can be exploited by adding bioemulsifiers and biosurfactants into environments where bioremediation/biodegradation rates of organic pollutants is to be enhanced. However, analysis of the current literature show some cases where the complex interactions among SACs, microbial cells, organic substrates and environmental media led to an inhibition of the biodegradation. The understanding of the different physiological roles of SACs in microbial communities is fundamental in order to develop more effective remediation technologies exploiting both synthetic surfactants and microbial SACs. The physio-chemical properties of some microbial SACs have been exploited in hydrocarbon-contaminated soils washing and in mobilisation of soil-bound metal in metal-contaminated soils. Our ability to analyse the microbial diversity in the natural environments will expand our knowledge on microbial SACs with respect to their exploitation for commercial applications and their roles in the physiology of the producing microorganisms.

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.

Season linked responses to fine and quasi-ultrafine Milan PM in cultured cells

Exposure to urbane airborne particulate matter (PM) is related to the onset and exacerbation of cardiovascular and respiratory diseases. The fine (PM1), and quasi-ultrafine (PM0.4) Milan particles collected during different seasons have been characterised and the biological effects on human epithelial lung A549, monocytes THP-1 cells and their co-culture, evaluated and compared with the results obtained on the PM10 and PM2.5 fractions. Chemical composition and transmission electron microscopy (TEM) analysis of PM0.4 showed that this fraction was very similar to PM1 for biological responses and dimension. All the winter fractions increased within 1h the level of reactive oxygen species (ROS), while only summer PM2.5 had this effect on A549 cells. The phosphorylation of H2AX (γH2AX), a marker of double strand DNA breaks (DSBs), was increased by all the winter fractions on A549 and THP-1 cells while summer PM samples did not induced this effect. PM0.4 and PM1 biological effects are partly similar and related to the season of sampling, with effects on ROS and DNA damage induced only by winter PM fractions. The winter PM damaging effect on DNA correlates with the presence of organic compounds.

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.

Thermophilic bacteria in cool temperate soils: are they metabolically active or continually added by global atmospheric transport?

Thermophilic soil geobacilli isolated from cool temperate geographical zone environments have been shown to be metabolically inactive under aerobic conditions at ambient temperatures (−5 to 25°C). It is now confirmed that a similar situation exists for their anaerobic denitrification activity. It is necessary therefore to determine the mechanisms that sustain the observed significant viable populations in these soils. Population analysis of thermophiles in rainwater and air samples has shown different species compositions which support the view that long distance global transport and deposition in rainwater is a possible source of replenishment of the soil thermophile populations. Survival experiments using a representative Geobacillus isolate have indicated that while cells lose viability rapidly at most temperatures, populations can increase only when the temperature allows growth to take place at a rate which exceeds death rate. Long term (9-month) experiments at 4°C show population increases which can be accounted for by very slow growth rates complemented by negligible death rates. These results are interpreted in the context of current hypotheses on the biogeography patterns of bacteria.

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.