Chiara Pesciaroli

Efficient removal of pollutants from olive washing wastewater in bubble-column bioreactor by Trametes versicolor

The white-rot fungi Panus tigrinus, Funalia trogii and Trametes versicolor have been tested in shake flasks for the reduction of olive washing wastewater (OWW) pollutants and production of oxidases on OWW-based media. P. tigrinus was rejected for its scarce performance. F. trogii showed best production of laccase (27 000 Ug(-1)), while T. versicolor appeared a good pollutant degrader reducing colour, COD and phenols by 60, 72 and 87%, respectively. Only T. versicolor grew well in bubble-column bioreactor: its OWW depollution, in continuous process, led to colour, COD and phenols reduction by 65%, 73% and 89%, respectively. Optimal dilution rate was 0.225d(-1) (0.225 m(3) of effluent treated daily per m(3) of bioreactor). Thus, a small bioreactor (10 m(3)) could treat daily the amount of OWW produced by a standard olive washing machine (2m(3)d(-1)). For these reasons, this process could be proposed as a simple, efficient and low-cost OWW treatment.

Temperature preferences of bacteria isolated from seawater collected in Kandalaksha Bay, White Sea, Russia

Fifty-two bacteria were isolated from seawater collected in Kandalaksha Bay, White Sea, Russia, and classified by 16S rDNA sequencing. Most of the strains belonged to ubiquitous microorganisms. Pseudomonas was the most abundant genus (21 strains), including species of P. fluorescens, P. putida and P. syringae. Serratia was also common (10 strains) with species S. plymuthica and S. proteamaculans. Sphingobacterium, Flavobacterium and Pantoea were less represented (5, 3 and 2 strains, respectively). The only typical bacterium of marine Arctic regions was Shewanella baltica. The strains were tested for their optimal growth temperature in the range 0–45°C. The majority appeared to be psychrotolerant (42%) or mesophilic-psychrotolerant (40%). In addition, one strain (Bacillus pumilus) showed a rather narrow mesophilic profile. No true psychrophilic bacteria were found. Most of the strains showed a classical curve with fast growth decrease above the optimum; some others displayed uncommon flat curves with scarce differences between maximum and minimum of growth in a wide range of temperatures. Moreover, few strains presented an unusual profile being, in relation to the optimum, more tolerant to high rather than low temperatures. Preferences of the Kandalaksha Bay strains are generally different from those reported in literature for the same species: optima were at lower temperatures and, sometimes, ranges were broader showing increased eurythermism. This could indicate adaptation to the wide temperature variations recorded in this peculiar environment.

Immobilization of Delftia tsuruhatensis in macro-porous cellulose and biodegradation of phenolic compounds in repeated batch process

Delftia tsuruhatensis BM90, previously isolated from Tyrrhenian Sea and selected for its ability to degrade a wide array of phenolic compounds, was immobilized in chemically modified macro porous cellulose. The development of bacterial adhesion on the selected carrier was monitored by scanning electron microscopy. Evident colonization started already after 8h of incubation. After 72h, almost all the carrier surface was covered by the bacterial cells. Extracellular bacterial structures, such as pili or fimbriae, contributed to carrier colonization and cell attachment. Immobilized cells of D. tsuruhatensis were tested for their ability to biodegrade a pool of 20 phenols in repeated batch process. During the first activation batch (72h), 90% of phenols degradation was obtained already in 48h. In the subsequent batches (up to 360h), same degradation was obtained after 24h only. By contrast, free cells were slower: to obtain almost same degradation, 48h were needed. Thus, process productivity, achieved by the immobilized cells, was double than that of free cells. Specific activity was also higher suggesting that the use of immobilized D. tsuruhatensis BM90 could be considered very promising in order to obtain an efficient reusable biocatalyst for long-term treatment of phenols containing effluents.

Bacterial community structure of a coastal area in Kandalaksha Bay, White Sea, Russia: possible relation to tidal hydrodynamics

Kandalaksha Bay is an estuarine system located around the North Polar Circle in the White Sea (Russia). This peculiar environment, showing big sea level differences during tide cycles causing intense water mixing, is almost unknown concerning its microbial diversity. In this work, seawater bacterial communities, mainly obtained from a coastal area, were studied in order to gather information on their structure and most abundant populations. The study was carried out by cluster analysis of polymerase chain reaction–temperature-gradient gel electrophoresis (PCR-TGGE) fingerprinting of partial 16S-rRNA gene amplicons. Bacterial communities were strongly homogenized by tidal water mixing, especially on surface layers and close to the shore. Samples collected from the intertidal zone and the nearby sea surface grouped together with a high percentage of similarity, while those taken offshore at various depths showed evident differences. Multivariate analysis indicated depth as the most significant environmental parameter causing variations in the community structure. High levels of diversity were revealed by both the Simpson’s index of diversity and the range-weighted richness index. The functional organization index suggested that the community was potentially able to preserve its functionality under stressing environmental perturbations. Sequencing of TGGE bands showed that most of the bacteria populations were evolutionarily close to α-proteobacteria. Some γ-proteobacteria and Actinobacteria were revealed too. This work represents the first major contribution to understanding bacterial diversity in Kandalaksha Bay.

Relationship between phylogenetic and nutritional diversity in Arctic (Kandalaksha Bay) seawater planktonic bacteria

Due to huge yearly variations of environmental stressing conditions, Kandalaksha Bay (Arctic Circle, White Sea, Russia) could represent a model to study microbial adaptation in extreme environments. This peculiar estuarine system has been scarcely investigated for its microbial diversity. In this work, to gather information on their nutritional competencies, seawater planktonic bacteria were studied for their ability to use different carbon sources by the Biolog phenotype microarray assays. Nestedness, a useful statistical tool used in ecology, was employed to underline nutritional differences among microbial groups. In particular, nestedness was used to understand the complex relationship that is established when many nutrients are available for various microorganisms, and to highlight presence of specialists and generalists. Among the studied bacteria, which showed very diverse nutritional abilities, 47% belonged to Pseudomonas, 21% to Serratia and 32% to other Genera. Within Pseudomonas, both highly generalist and highly specialist strains were discovered. However, most of them used organic and/or amino acids as principal carbon sources. In contrast, Serratia strains typically preferred sugars and appeared to be more generalist. On the whole, important differences in specialization levels and nutritional competencies were recorded in strains belonging to the same species. Correlations between phylogenetic and nutritional data were validated by Procrustes analysis.

Effect of semi-permeable cover system on the bacterial diversity during sewage sludge composting

Sewage sludge composting is a profitable process economically viable and environmentally friendly. In despite of there are several kind of composting types, the use of combined system of semipermeable cover film and aeration air-floor is widely developed at industrial scale. However, the knowledge of the linkages between microbial communities structure, enzyme activities and physico-chemical factors under these conditions it has been poorly explored. Thus, the aim of this study was to investigate the bacterial dynamic and community structure using next generation sequencing coupled to analyses of microbial enzymatic activity and culturable dependent techniques in a full-scale real composting plant. Sewage sludge composting process was conducted using a semi-permeable Gore-tex cover, in combination with an air-insufflation system. The highest values of enzymatic activities such as dehydrogenase, protease and arylsulphatase were detected in the first 5 days of composting; suggesting that during this period of time a greater degrading activity of organic matter took place. Culturable bacteria identified were in agreement with the bacteria found by massive sequencing technologies. The greatest bacterial diversity was detected between days 15 and 30, with Actinomycetales and Bacillales being the predominant orders at the beginning and end of the process. Bacillus was the most representative genus during all the process. A strong correlation between abiotic factors as total organic content and organic matter and enzymatic activities such as dehydrogenase, alkaline phosphatase, and ß-glucosidase activity was found. Bacterial diversity was strongly influenced by the stage of the process, community-structure change was concomitant with a temperature rise, rendering favorable conditions to stimulate microbial activity and facilitate the change in the microbial community linked to the degradation process. Moreover, results obtained confirmed that the use of semipermeable cover in the composting of sewage sludge allow a noticeable reduction in the process-time comparing to conventional open windrows.

Linking the Effect of Antibiotics on Partial-Nitritation Biofilters: Performance, Microbial Communities and Microbial Activities

The emergence and spread of antibiotics resistance in wastewater treatment systems have been pointed as a major environmental health problem. Nevertheless, research about adaptation and antibiotics resistance gain in wastewater treatment systems subjected to antibiotics has not been successfully developed considering bioreactor performance, microbial community dynamics and microbial activity dynamics at the same time. To observe this in autotrophic nitrogen removal systems, a partial-nitritation biofilter was subjected to a continuous loading of antibiotics mix of azithromycin, norfloxacin, trimethoprim, and sulfamethoxazole. The effect of the antibiotics mix over the performance, bacterial communities and bacterial activity in the system was evaluated. The addition of antibiotics caused a drop of ammonium oxidation efficiency (from 50 to 5%) and of biomass concentration in the bioreactor, which was coupled to the loss of ammonium oxidizing bacteria Nitrosomonas in the bacterial community from 40 to 3%. Biomass in the partial nitritation biofilter experienced a sharp decrease of about 80% due to antibiotics loading, but the biomass adapted and experienced a growth by stabilization under antibiotics feeding. During the experiment several bacterial genera appeared, such as Alcaligenes, Paracoccus, and Acidovorax, clearly dominating the bacterial community with >20% relative abundance. The system reached around 30% ammonium oxidation efficiency after adaptation to antibiotics, but no effluent nitrite was found, suggesting that dominant antibiotics-resistant phylotypes could be involved in nitrification–denitrification metabolisms. The activity of ammonium oxidation measured as amoA and hao gene expression dropped a 98.25% and 99.21%, respectively, comparing the system before and after the addition of antibiotics. On the other hand, denitrifying activity increased as observed by higher expression of nir and nos genes (83.14% and 252.54%, respectively). In addition, heterotrophic nitrification cyt c-551 was active only after the antibiotics addition. Resistance to the antibiotics was presumably given by ermF, carA and msrA for azithromycin, mutations of the gyrA and grlB for norfloxacin, and by sul123 genes for sulfamethoxazole. Joined physicochemical and microbiological characterization of the system were used to investigate the effect of the antibiotics over the bioprocess. Despite the antibiotics resistance, activity of Bacteria decreased while the activity of Archaea and Fungi increased.

Pollutants degradation performance and microbial community structure of aerobic granular sludge systems using inoculums adapted at mild and low temperature

Three aerobic granular sequencing batch reactors were inoculated using different inocula from Finland, Spain and a mix of both in order to investigate the effect over the degradation performance and the microbial community structure. The Finnish inoculum achieved a faster granulation and a higher depollution performance within the first two month of operation. However, after 90 days of operation, similar physico-chemical values were observed. On the other hand, the Real-time PCR showed that Archaea diminished from inoculum to granular biomass, while Bacteria and Fungi numbers remained stable. All granular biomass massive parallel sequencing studies were similar regardless of the inocula from which they formed, as confirmed by singular value decomposition principal coordinates analysis, expected effect size of OTUs, and β-diversity analyses. Thermoproteaceae, Meganema and a Trischosporonaceae members were the dominant phylotypes for the three domains studied. The analysis of oligotype distribution demonstrated that a fungal oligotype was ubiquitous. The dominant OTUs of Bacteria were correlated with bioreactors performance. The results obtained determined that the microbial community structure of aerobic granular sludge was similar regardless of their inocula, showing that the granulation of biomass is related to several phylotypes. This will be of future importance for the implementation of aerobic granular sludge to full-scale systems.