Tannery Wastewater Recalcitrant Compounds Foster the Selection of Fungi in Non-Sterile Conditions: A Pilot Scale Long-Term Test.

This study demonstrated that a microbial community dominated by fungi can be selected and maintained in the long-term under non-sterile conditions, in a pilot-scale packed-bed reactor fed with tannery wastewater. During the start-up phase, the reactor, filled with 0.6 m3 of polyurethane foam cubes, was inoculated with a pure culture of Aspergillus tubingensis and Quebracho tannin, a recalcitrant compound widely used by tannery industry, was used as sole carbon source in the feeding. During the start-up, fungi grew attached as biofilm in carriers that filled the packed-bed reactor. Subsequently, the reactor was tested for the removal of chemical oxygen demand (COD) from an exhaust tanning bath collected from tanneries. The entire experiment lasted 121 days and average removals of 29% and 23% of COD and dissolved organic carbon (DOC) from the tannins bath were achieved, respectively. The evolution of the microbial consortium (bacteria and fungi) was described through biomolecular analyses along the experiment and also developed as a function of the size of the support media.

Role of bacterivorous organisms on fungal-based systems for natural tannin degradation.

Tannery wastewater presents high concentrations of organic load and pollutant recalcitrant molecules (e.g. tannins), which reduce the efficiency of biological treatment processes. Recent studies showed that several fungal species and strains are effective in the degradation of tannins. However, high bacterial load can negatively affect fungal growth, reducing system stability and degradation performances. The aim of the present study was to evaluate the effects of the introduction of bacterivorous grazers (ciliates and/or rotifers) in batch scale experiments using fungi to remove Tara tannin, i.e. to check the potential synergistic effect between fungi and bacterivorous grazers in the degradation of recalcitrant compounds. In this context, the ciliated grazers Paramecium calkinsi, Tetrahymena sp., Pseudovorticella sp., and the rotifer Lecane inermis, preliminary selected according to their ability to grow in a solution prepared with Tara tannin, were separately tested. Activated sludge, including a complex mixture of native grazers, was used as experimental control. The following parameters were monitored: bacterial load, number of grazers/mL and Soluble Chemical Oxygen Demand (SCOD). Colony Forming Unit (CFU)/grazers ratio was also calculated. Particular attention was paid to: i) bacterial load reduction and ii) enhancement of recalcitrant compounds degradation, and we observed that in all experimental conditions where grazers occurred bacterial load was significantly reduced and the system achieved a higher SCOD removal in a shorter time. Our findings provide useful insights for the stabilization of fungal-based systems in non-sterile conditions.

The role of cosubstrate and mixing on fungal biofilm efficiency in the removal of tannins.

Tannins are polyphenolic compounds produced by plants and they are used in industrial vegetable tanning of leather. Tannins represent one of the low biodegradability substances in tannery wastewaters with high recalcitrant soluble chemical oxygen demand, furthermore high concentration of tannins can inhibit biological treatment. In the present study, four novel rotating submerged packed bed reactors were inoculated with a selected fungal strain to reach a biological degradation of tannins in non-sterile conditions. The selected fungal strain, Aspergillus tubingensis MUT 990, was immobilised in polyurethane foam cubes carriers and inserted inside a submerged rotating cage reactors. The reactors were feed with a solution composed of four tannins: Quebracho (Schinopsis spp.), Wattle (Mimosa spp.), Chestnut (Castanea spp.) and Tara (Caesalpinia spp.). Four reactors with a volume of 4 L each were used, the co-substrate was pure malt extract, the hydraulic retention time was 24 h and the pH setpoint was 5.5. The reactors configuration was chosen to allow the study of the effect of rotation and the co-substrate addition on tannins removal. The experiment lasted two months and it was achieved 80% of chemical oxygen demand and up to 90% dissolved organic carbon removal, furthermore it was detected an important tannase activity.

The microbial community in a moving bed biotrickling filter operated to remove hydrogen sulfide from gas streams.

The information available on the microbial communities responsible for pollutant degradation is increasingly accessible. Its use to optimize process design and operation is an important challenge in the field of effluent treatment research. Therefore, a prototype of a moving bed biotrickling filter (MBBTF) reactor was designed and, for the first time, operated at full-scale for the removal of sulfides desorbing from tannery industrial wastewater. The bacterial community operating in this innovative reactor was studied, and its composition and response to different operating conditions were characterized. A stable biomass, dominated by sulfur-oxidizing bacteria of the genus Acidithiobacillus was selected from inside the MBBTF reactor, and temperature, pH and bed rotation were shown to be the main factors driving the community structure. Moreover, data from different approaches indicated an uneven spatial distribution of biofilm inside the studied reactor, due to the combined effect of fluid dynamics and substrate gradients within the bed volume. Despite the high removal efficiency achieved by this innovative prototype (80% on average), the data suggested that the result could be improved by adopting solutions for a more stable and even biofilm distribution. It was shown that short frequent bed rotations, rather than long scattered rotations, ensured biomass stability. Furthermore, diversifying biofilm support media as a function of expected local pollutant concentrations should be considered. Data obtained from the bacterial community can therefore provide indications for possible further improvement of MBBTF reactor design and performance.

Summer holidays as break-point in shaping a tannery sludge microbial community around a stable core microbiota.

Recently, several investigations focused on the discovery of a bacterial consortium shared among different wastewater treatment plants (WWTPs). Nevertheless, the definition of a core microbiota over time represents the necessary counterpart in order to unravel the dynamics of bacterial communities in these environments. Here we performed a monthly survey on the bacterial community of a consortial industrial plant. Objectives of this study were: (1) to identify a core microbiota constant over time; (2) to evaluate the temporal dynamics of the community during one year. A conspicuous and diversified core microbiota is constituted by operational taxonomic units which are present throughout the year in the plant. Community composition data confirm that the presence and abundance of bacteria in WWTPs is highly consistent at high taxonomic level. Our results indicate however a difference in microbial community structure between two groups of samples, identifying the summer holiday period as the break-point. Changes in the structure of the microbial community occur otherwise gradually, one month after another. Further studies will clarify how the size and diversity of the core microbiota could affect the observed dynamics.

Nitrifying biomass characterization and monitoring during bioaugmentation in a membrane bioreactor.

A membrane bioreactor (MBR), fed with domestic wastewater, was bioaugmented with nitrifying biomass selected in a side-stream MBR fed with a synthetic high nitrogen-loaded influent. Microbial communities evolution was monitored and comparatively analysed through an extensive bio-molecular investigation (16S rRNA gene library construction and terminal-restriction fragment length polymorphism techniques) followed by statistical analyses. As expected, a highly specialized nitrifying biomass was selected in the side-stream reactor fed with high-strength ammonia synthetic wastewater. The bioaugmentation process caused an increase of nitrifying bacteria of the genera Nitrosomonas (up to more than 30%) and Nitrobacter in the inoculated MBR reactor. The overall structure of the microbial community changed in the mainstream MBR as a result of bioaugmentation. The effect of bioaugmentation in the shift of the microbial community was also verified through statistical analysis.

Modeling bioaugmentation with nitrifiers in membrane bioreactors.

Bioaugmentation with nitrifiers was studied using two pilot-scale membrane bioreactors, with the purpose of assessing the suitability of state-of-the-art activated sludge models (ASMs) in predicting the efficiency of bioaugmentation as a function of operating conditions. It was demonstrated that the temperature difference between seeding and seeded reactors (ΔT) affects bioaugmentation efficiency. Experimental data were accurately predicted when ΔT was within a range of up to 10 °C at the higher range, and when the temperature was significantly lower in the seeded reactor compared to the seeding one, standard ASMs overestimated the efficiency of bioaugmentation. A modified ASM, capable of accurately representing the behavior of seeded nitrifying biomass in the presence of high ΔT, would require the inclusion of the effect of temperature time gradients on nitrifiers. A simple linear correlation between ΔT and the Arrhenius coefficient was proposed as a preliminary step.

Characterization and comparison of bacterial communities selected in conventional activated sludge and membrane bioreactor pilot plants: a focus on Nitrospira and Planctomycetes bacterial phyla.

A pilot-scale membrane bioreactor (MBR) and a conventional activated sludge system (CAS) were in parallel operated to investigate the impact of the separation technology on the structure and functionality of the selected microbial community. Microbial communities as well as nitrogen removal efficiency of the biomass were characterized. Kinetics and microbial community structure turned out to be duly correlated. The impact of the separation technology on selective conditions and, in particular, the higher variability of solid separation efficiency in CAS with respect to MBR pilot plant possibly represented the main factor influencing the selection of bacterial communities. Concerning nitrifiers, bacteria of the genus Nitrospira were predominant in the MBR. This was in accordance with kinetics of nitrite-oxidizing bacteria that suggested the presence of k-strategists, while r-strategists were selected in the CAS plant, possibly because of the presence of transient higher concentrations of nitrite (in the range of 0.05-0.18 and of 0.05-4.4 mg [Formula: see text]-N L(-1) in the MBR and CAS effluents, respectively). An unexpectedly high presence of bacteria belonging to two specific phylogenetic clades of Planctomycetes was found in both reactors.

Biomass accumulation modelling in a highly loaded biotrickling filter for hydrogen sulphide removal.

A pilot scale test on a biotrickling filter packed with polyurethane foam cubes was carried out for 110 d at high volumetric mass load (up to 280 g m(bed)(-3) h(-1)) with the aim of studying the accumulation of solids in the treatment of H(2)S. Removal rate up to 245 g m(bed)(-3) h(-1) was obtained; however, an accumulation of gypsum, elemental sulphur and, above all, inert biomass was identified as the cause of an increased pressure drop over the long term. A mathematical model was applied and calibrated with the experimental results to describe the accumulation of biomass. The model was capable of describing the accumulation of solids and, corresponding to a solids retention time of 50 d, the observed yield resulted in 0.07 g of solids produced g(-1) H(2)S removed. Respirometric tests showed that heterotrophic activity is inhibited at low pH (pH < 2.3), and the contribution to biomass removal through decay was negligible.

Monitoring biological sulphide oxidation processes using combined respirometric and titrimetric techniques.

The application of respirometric and titrimetric techniques to evaluate kinetic parameters and stoichiometry of the sulphide-oxidising biomass is a new promising approach for biotechnological sulphide oxidation process monitoring. It was possible to estimate the yield coefficients of each oxidation step of sulphide to elemental sulphur and to sulphate using respirometric tests, while evaluating the behaviour of the biomass in endogenous conditions. Furthermore, it was demonstrated how the combined application of titrimetric and respirometric techniques enabled the monitoring of sulphur and sulphate formation as a function of the environmental conditions. This approach provided valuable information of the biological sulphide oxidation processes and preliminary results may be used as a starting point for the formulation and use of a mathematical model.

Sulphide oxidation to elemental sulphur in a membrane bioreactor: performance and characterization of the selected microbial sulphur-oxidizing community.

In leather tanning industrial areas sulphide management represents a major problem. However, biological sulphide oxidation to sulphur represents a convenient solution to this problem. Elemental sulphur is easy to separate and the process is highly efficient in terms of energy consumption and effluent quality. As the oxidation process is performed by specialized bacteria, selection of an appropriate microbial community is fundamental for obtaining a good yield. Sulphur oxidizing bacteria (SOB) represent a wide-ranging and highly diversified group of microorganisms with the capability of oxidizing reduced sulphur compounds. Therefore, it is useful to select new microbes that are able to perform this process efficiently. For this purpose, an experimental membrane bioreactor for sulphide oxidation was set up, and the selected microbial community was characterized by constructing 16S rRNA gene libraries and subsequent screening of clones. Fluorescence in situ hybridization (FISH) was then used to assess the relative abundance of different bacterial groups. Sulphide oxidation to elemental sulphur proceeded in an efficient (up to 79% conversion) and stable way in the bioreactor. Both analysis of clone libraries and FISH experiments revealed that the dominant operational taxonomic unit (OTU) in the bioreactor was constituted by Gammaproteobacteria belonging to the Halothiobacillaceae family. FISH performed with the specifically designed probe tios_434 demonstrated that this OTU constituted 90.6+/-1.3% of the bacterial community. Smaller fractions were represented by bacteria belonging to the classes Betaproteobacteria, Alphaproteobacteria, Deltaproteobacteria, Clostridia, Mollicutes, Sphingobacteria, Bacteroidetes and Chlorobia. Phylogenetic analysis revealed that clone sequences from the dominant OTU formed a stable clade (here called the TIOS44 cluster), within the Halothiobacillaceae family, with sequences from many organisms that have not yet been validly described. The data indicated that bacteria belonging to the TIOS44 cluster were responsible for the oxidation process.