Nitrification activity in the presence of 2-chlorophenol using whole nitrifying cells and cell-free extracts: batch and SBR assays.

Kinetic assays with a nitrifying consortium with whole nitrifying cells amended with 5 mg 2-CP-C/L and 100, 200, 300, or 500 mg NH4+-N/L were carried out in batch and nitrifying sequencing batch reactor (SBR) cultures. No nitrification activity was observed in batch assays with 100 mg NH4+-N/L and 5 mg 2-CP-C/L. Nevertheless, increasing the ammonium concentration from 200 to 500 mg NH4+-N/L allowed simultaneous ammonium and nitrite oxidation even in the presence of 5 mg 2-CP-C/L plus the halogenated compound consumption. Under these conditions, the ammonium monooxygenase enzyme participated in 2-CP consumption. Complete nitrification and simultaneous elimination of 5 mg 2-CP-C/L were achieved in the SBR amended with 200-500 mg NH4+-N/L. The inhibitory effect of 2-CP on the nitrite oxidation process completely disappeared under these conditions. Assays with nitrifying cell-free extracts, ammonium (100 mg NH4+-N/L), and 2-CP (5 mg 2-CP-C/L) were also conducted. In the absence of 2-CP, the nitrifying cell-free extracts maintained up to 60% of the nitrifying activity compared to whole-cells. Contrary to whole-cell assays, cell-free extracts were capable of simultaneously oxidizing ammonium and consuming 2-CP. However, the inhibitory effect of 2-CP on nitrification was still present as lower specific rates of ammonium consumption and nitrate production were obtained. Thus, these assays indicate that the presence of 2-CP affects both, the ammonium transport mechanism and the activity of nitrifying enzymes. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03764-z.

Kinetic and physiological evaluation of ammonium and nitrite oxidation processes in presence of 2-chlorophenol.

The effect of 2-chlorophenol (2-CP) on ammonium and nitrite-oxidizing processes was kinetically evaluated in batch cultures with nitrifying sludge at steady state. Assays with ammonium or nitrite as energy source and 2.5, 5.0, or 10.0 mg 2-CP-C/l were conducted. Control assays without 2-CP were also performed. Ammonium-oxidizing activity was completely inhibited at the different 2-CP concentrations, whereas nitrite-oxidizing activity was present as nitrite was completely consumed and converted to nitrate irrespectively of 2-CP concentration. In the presence of 2.5 and 5.0 mg 2-CP-C/l, no significant effect on specific rates of nitrite consumption and nitrate production was observed, but a significant decrease on these parameters was observed at 10.0 mg 2-CP-C/l. The nitrifying sludge previously exposed to 2-CP was unable to completely recover its ammonium and nitrite oxidation capacity. Nevertheless, complete 2-CP consumption was achieved in all assays. The effect of 2-CP on ammonium oxidation was observed at kinetic and metabolic pathway level, whereas the effect on nitrite oxidation was observed only at kinetic level. The results obtained in this work evidenced that in order to achieve a successful nitrification process the presence in wastewater of even 2.5 mg 2-CP-C/l should be avoided.

p-Cresol biotransformation by a nitrifying consortium.

The oxidizing ability of a nitrifying consortium exposed to p-cresol (25 mg CL(-1)) was evaluated in batch cultures. Biotransformation of the phenolic compound was investigated by identifying the different intermediates formed. p-Cresol inhibited the ammonia-oxidizing process with a decrease of 83% in the specific rate of ammonium consumption. After 48 h, ammonium consumption efficiency was 96+/-9% while nitrate yield reached 0.95+/-0.06 g NO(3)(-)-Ng(-1)NH(4)(+)-N consumed. High value for nitrate production yield showed that the nitrifying metabolic pathway was only affected at the specific rate level being nitrate the main end product. The consortium was able to totally oxidize p-cresol at a specific rate of 0.17+/-0.06 mg p-cresol-Cmg(-1) microbial protein h(-1). p-Cresol was first transformed to p-hydroxybenzaldehyde and p-hydroxybenzoate, which were later completely mineralized. In the presence of allylthiourea, a specific inhibitor of ammonia monooxygenase (AMO), p-cresol was oxidized to the same intermediates and in a similar pattern as obtained without the AMO inhibitor. AMO seemed not to be involved in the p-cresol oxidation process. When p-hydroxybenzaldehyde was added (25 mg CL(-1)), the nitrifying process was inhibited in the same way as observed with p-cresol, indicating that p-hydroxybenzaldehyde could be the main compound responsible for nitrification inhibition. p-Hydroxybenzaldehyde was accumulated during 15 h before complete consumption at a specific rate value eight times lower than the p-cresol consumption rate. Results showed that p-hydroxybenzaldehyde oxidation was the limiting step in p-cresol mineralization by the nitrifying consortium.

Qualitative and quantitative determination of a humic model compound in microbial cultures by cyclic voltammetry.

The humic model compound, anthraquinone-2,6-disulfonate (AQDS), was characterized and measured in microbial cultures by cyclic voltammetry (CV). Under the experimental conditions, the formal reduction potential (E(o')) of the couple AQDS/AHQDS was found to be of -0.520 V vs. SCE (standard calomel electrode) at pH value of 7.0. Control experiments showed that there were no interferences of the culture medium or the microbial consortium on the quantitative determination of the quinone. The linear equation E(o') = -0.294 - 0.032 pH was found, showing that the pH used (7.0-7.8) did not affect significantly the AQDS determination by CV and AHQDS was the predominant hydroquinone formed. A calibration curve was obtained by plotting current response versus AQDS concentration with a linear correlation (r = 0.999) from 0.2 to 10 mM of AQDS. This technique was applied in a denitrifying culture to establish kinetic profiles for AHQDS formation coupled to acetate and p-cresol oxidation. CV results showed that organic matter oxidation by the denitrifying sludge was stoichiometrically associated to AQDS reduction into AHQDS-. CV was shown to be a useful tool for monitoring oxidation/reduction processes of quinones occurring in complex microbial media.

Batch nitrifying cultures in presence of mixtures of benzene, toluene, and m-xylene.

Benzene, toluene, and m-xylene compounds in individual (5.0 +/- 0.5 mg C l(-1)) and mixed solutions (2.5 +/- 0.2 mg C l(-1) for each one) in nitrifying batch cultures induced a decrease in the specific rates of NH4+ consumption (81 +/- 6%) and NO3- production (39-79%). However, after 24 h, ammonium consumption efficiency and conversion of consumed NH4+ -N into NO3- -N were close to 100% and there was no significant accumulation of nitrite in the medium. After 24 h, no aromatic intermediate was detected in the cultures and 50% of the mixed compounds was converted to acetate. The following order of biotransformation was found: m-xylene > toluene > benzene. Transformation rate of m-xylene was 0.051 +/- 0.005 g C (g protein-N h)(-1) in individual and mixed solutions. When m-xylene was added, benzene was oxidized at a faster rate (0.051 +/- 0.005 g C (g protein-N h)(-1)) whereas toluene at a slower rate (0.012 +/- 0.002 g C (g protein-N h)(-1)).

Simultaneous nitrification and p-cresol oxidation in a nitrifying sequencing batch reactor.

The tolerance, kinetic behavior and oxidizing ability of a nitrifying sludge exposed to different initial concentrations of p-cresol (25-150mg/l) were evaluated in a sequencing batch reactor (SBR) fed with 200mg NH(4)(+)-N/ld. The nitrifying SBR operated up to 300mg/ld of p-cresol, achieving simultaneously the complete ammonium oxidation to nitrate and the total consumption of p-cresol and its transitory intermediates from the culture. p-Cresol induced a significant decrease in the values for specific rates of ammonium consumption, showing that the ammonium oxidation pathway was mainly inhibited. After 7 months of operation in SBR, the specific rates of NH(4)(+)-N oxidation, NO(3)(-)-N formation, and total organic carbon consumption were 0.6g NH(4)(+)-N/g microbial protein-Nh, 0.3g NO(3)(-)-N/g microbial protein-Nh, and 0.24g total organic carbon/g microbial protein h, respectively. The microbial growth rate was always low (maximum value of 12.2+/-0.4mg protein-N/ld) and settleability of the sludge was good with sludge volume index values lower than 21ml/g. The oxidation of p-cresol and its intermediates was carried out faster throughout the cycles and nitrification inhibition decreased with the number of cycles.

Kinetic and metabolic study of benzene, toluene and m-xylene in nitrifying batch cultures.

The effect of benzene, toluene, and m-xylene (BTX) compounds on the nitrifying activity of a sludge produced in steady-state nitrification was evaluated in batch cultures. Benzene and m-xylene at 10 mg C/L decreased ammonium consumption efficiency by 57% and 26%, respectively, whereas toluene did not affect the ammonium oxidation process. The consumed NH4+-N was totally oxidized to NO3- -N. There was no significant effect at 5 mg C/L of each aromatic compound. BTX (5-20mg C/L) induced a significant decrease in the values for specific rates of NH4+ -N consumption (76-99%) and NO3- -N production (45-98%). At 10 mg C/L of BTX compounds, the inhibition order on nitrate production was: benzene > m-xylene > toluene while at 20 mg C/L, the sequence changed to m-xylene > toluene > benzene for both nitrification inhibition and BTX compounds persistence. At 5 mg C/L of BTX compounds, there was no toxic effect on the sludge whereas from 10 to 50 mgC/L, bacteria did not totally recover their nitrifying activity. At a concentration of 5 mg C/L, toluene was first oxidized to benzyl alcohol, which was later oxidized to butyrate while m-xylene was oxidized to acetate and butyrate.

Settleability and kinetics of a nitrifying sludge in a sequencing batch reactor.

A physiological study of a nitrifying sludge was carried out in a sequencing batch reactor (SBR). Pseudo steady-state nitrification conditions were obtained with an ammonium removal efficiency of 99% +/- 1% and 98% +/- 2% conversion of NH4+-N to NO3 - -N. The rate of biomass production was negligible (1.3 +/- 0.1 mg microbial protein-N.L(-1).d(-1)). The sludge presented good settling properties with sludge volume index values lower than 20 mL.g(-1) and an exopolymeric protein/carbohydrate ratio of 0.53 +/- 0.34. Kinetic results indicated that the nitrifying behavior of the sludge changed with the number of cycles. After 22 cycles, a decrease in the specific rate of NO3- -N production coupled with an increase in the NO2- -N accumulation were observed. These results showed that the activity of the nitrite oxidizing bacteria decreased at a longer operation time. Ammonia oxidizing bacteria were found to exhibit the best stability. After 4 months of operation, the specific rates of NH4+-N consumption and NO3- -N production were 1.72 NH4+-N per microbial protein-N per hour (g.g(-1).h(-1)) and 0.54 NO3- -N per microbial protein-N per hour (g.g(-1).h(-1)), respectively.

Benzene transformation in nitrifying batch cultures.

The effect of benzene on the nitrifying activity of a sludge produced in steady-state nitrification was evaluated in batch cultures. Benzene at 10 mg/L inhibited nitrate formation by 53%, whereas at 5 mg/L there was no inhibition. For initial benzene concentrations of 0, 7, and 10 mg/L, the specific rates of NO(3)(-)-N production were 0.545 +/- 0.101, 0.306 +/- 0.024, and 0.141 +/- 0.010 g NO(3)(-)-N/g microbial protein-N.h, respectively. The specific rates of benzene consumption at 7, 12, and 20 mg/L were 0.034 +/- 0.003, 0.050 +/- 0.006, and 0.027 +/- 0.002 g/g microbial protein-N.h, respectively. Up to a concentration of 10 mg/L, benzene was first oxidized to phenol, which was later totally oxidized to acetate. Benzene at higher concentrations (20 and 30 mg/L) was converted to intermediates other than acetate, phenol, or catechol. These results suggest that this type of nitrifying consortium coupled with a denitrification system may have promising applications for complete removal of nitrogen and benzene from wastewaters.

Rare earth elements removal by microbial biosorption: a review.

This paper reviews published work on the sorption of rare earth elements by microbial biomass. In a first part, the biosorption capacities and the various experimental conditions performed in batch reactor experiments are compared. Secondly, sorption modelling generally used in biosorption studies are described. Thirdly, the microbial cell wall characteristics of the metallic ion binding sites are considered. From these observations it seems that the important functional groups for metallic ion fixation are the carboxyl and the phosphate moieties. Moreover, the competing effect of various ions like aluminium, iron, glutamate, sulphate etc. is described. Finally, some adsorption results of the rare earth elements in dynamic reactors are presented.

Fixed-bed study for lanthanide (La, Eu, Yb) ions removal from aqueous solutions by immobilized Pseudomonas aeruginosa: experimental data and modelization.

A fixed-bed study was carried out by using cells of Pseudomonas aeruginosa immobilized in polyacrylamide gel as a biosorbent for the removal of lanthanide (La, Eu, Yb) ions from aqueous solutions. The effects of superficial liquid velocity based on empty column, particle size, influent concentration and bed depth on the lanthanum breakthrough curves were investigated. Immobilized biomass effectively removed lanthanum from a 6 mM solution with a maximum adsorption capacity of 342 micromolg(-1) (+/-10%) corresponding closely to that observed in earlier batch studies with free bacterial cells. The Bohart and Adams sorption model was employed to determine characteristic parameters useful for process design. Results indicated that the immobilized cells of P. aeruginosa enable removal of lanthanum, europium and ytterbium ions from aqueous effluents with significant and similar maximum adsorption capacities. Experiments with a mixed cation solution showed that the sequence of preferential biosorption was Eu3+ > or = Yb3+ > La3+. Around 96+/-4% of the bound lanthanum was desorbed from the column and concentrated by eluting with a 0.1 M EDTA solution. The feasibility of regenerating and reusing the biomass through three adsorption/desorption cycles was suggested. Neural networks were used to model breakthrough curves performed in the dynamic process. The ability of this statistical tool to predict the breakthrough times was discussed.