Mineral characterization of the biogenic Fe(III)(hydr)oxides produced during Fe(II)-driven denitrification with Cu, Ni and Zn.

The recovery of iron and other heavy metals by the formation of Fe(III) (hydr)oxides is an important application of microbially-driven processes. The mineral characterization of the precipitates formed during Fe(II)-mediated autotrophic denitrification with and without the addition of Cu, Ni, and Zn by four different microbial cultures was investigated by X-ray fluorescence (XRF), Raman spectroscopy, scanning electron microscopy equipped with energy dispersive X-Ray analyzer (SEM-EDX), Fourier transform infrared spectroscopy (FTIR) and X-ray Powder Diffraction (XRD) analyses. Fe(II)-mediated autotrophic denitrification resulted in the formation of a mixture of Fe(III) (hydr)oxides composed of amorphous phase, poorly crystalline (ferrihydrite) and crystalline phases (hematite, akaganeite and maghemite). The use of a Thiobacillus-dominated mixed culture enhanced the formation of akaganeite, while activated sludge enrichment and the two pure cultures of T. denitrificans and Pseudogulbenkiania strain 2002 mainly resulted in the formation of maghemite. The addition of Cu, Ni and Zn led to similar Fe(III) (hydr)oxides precipitates, probably due to the low metal concentrations. However, supplementing Ni and Zn slightly stimulated the formation of maghemite. A thermal post-treatment performed at 650 °C enhanced the crystallinity of the precipitates and favored the formation of hematite and some other crystalline forms of Fe associated with P, Na and Ca.

Effect of Cu, Ni and Zn on Fe(II)-driven autotrophic denitrification.

Fe(II)-mediated autotrophic denitrification in the presence of copper (Cu), nickel (Ni) and zinc (Zn) with four different microbial cultures was investigated in batch bioassays. In the absence of metals, complete nitrate removal and Fe(II) oxidation were achieved with a Thiobacillus-dominated mixed culture and Pseudogulbenkiania sp. 2002 after 7 d. A nitrate removal of 96 and 91% was observed with a pure culture of T. denitrificans and an activated sludge enrichment, respectively, after 10 d of incubation. Cu, Ni and Zn were then supplemented at an initial concentration of 5, 10, 20 and 40 mg Me/L. A decrease of approximately 50% of the soluble metal concentrations occurred in the first 4 d of denitrification, due to metal precipitation, co-precipitation, sorption onto iron (hydr)oxides, and probably sorption onto biomass. A higher sensitivity to metal toxicity was observed for the microbial pure cultures. Pseudogulbenkiania sp. 2002 was the least tolerant among the biomasses tested, resulting in only 6, 8 and 6% nitrate removal for the highest Cu, Ni and Zn concentrations, respectively. In contrast, the highest nitrate removal efficiency and specific rates were achieved with the Thiobacillus-dominated mixed culture, which better tolerated the presence of metals. Averagely, Cu resulted in the highest inhibition of nitrate removal, followed by Zn and Ni.

Influence of pH, EDTA/Fe(II) ratio, and microbial culture on Fe(II)-mediated autotrophic denitrification.

Fe(II)-mediated autotrophic denitrification with four different microbial cultures under different pH and EDTA/Fe(II) conditions was investigated in batch bioassays. Initially, the highest nitrate removal (72%) was achieved with an activated sludge inoculum. The use of pure cultures of Pseudogulbenkiania strain 2002 and Thiobacillus denitrificans resulted in a 55 and 52% nitrate removal, respectively. No denitrification was observed for a mixed culture dominated by Thiobacillus thioparus and T. denitrificans. A longer enrichment on Fe(II) and the supplementation of thiosulfate as additional electron donor were needed to stimulate the denitrifying activity of the Thiobacillus-mixed culture. A second subculture on Fe(II) as sole electron donor resulted in higher denitrification efficiencies for all microbial cultures. In particular, nitrate removal reached up to 84% with a specific nitrate removal rate of 1.160 mM·(g VSS·day)-1 in the bioassays seeded with the Thiobacillus-mixed culture. All cultures were favored by decreasing the EDTA/Fe(II) molar ratio from 2.0 to 0.5. The most significant denitrification enhancement was observed for the Pseudogulbenkiania species, indicating a lower tolerance to EDTA. The two pure cultures effectively maintained denitrification at pH 7.0 and were more sensitive to a pH decrease. Conversely, the optimal pH was 6.0 for the Thiobacillus-mixed and activated sludge cultures.