Physiological adaptation and population dynamics of a nitrifying sludge exposed to ampicillin.

Antibiotics in wastewater treatment plants can alter the physiological activity and the structure of microbial communities through toxic and inhibitory effects. Physiological adaptation, kinetic, and population dynamics behavior of a nitrifying sludge was evaluated in a sequential batch reactor (SBR) fed with 14.4 mg/L of ampicillin (AMP). The addition of AMP did not affect ammonium consumption (100 mg NH4+-N/L) but provoked nitrite accumulation (0.90 mg NO2--N formed/mg NH4+-N consumed) and an inhibition of up to 67% on the nitrite oxidizing process. After 30 cycles under AMP feeding, the sludge recovered its nitrite oxidizing activity with a high nitrate yield (YNO3-) of 0.87 ± 0.10 mg NO3--N formed/mg NH4+-N consumed, carrying out again a stable and complete nitrifying process. Increases in specific rate of nitrate production (qNO3-) showed the physiological adaptation of the nitrite oxidizing bacteria to AMP inhibition. Ampicillin was totally removed since the first cycle of addition. Exposure to AMP had effects on the abundance of bacterial populations, promoting adaptation of the nitrifying sludge to the presence of the antibiotic and its consumption. Nitrosomonas and Nitrosospira always remained within the dominant genera, keeping the ammonium oxidizing process stable while an increase in Nitrospira abundance was observed, recovering the stability of the nitrite oxidizing process. Burkholderia, Pseudomonas, and Thauera might be some of the heterotrophic bacteria involved in AMP consumption.

Long-term post-storage reactivation of a nitrifying sludge in a sequential batch reactor: physiological and kinetic evaluation.

Production, preservation and recovery of sludge with stabilized nitrifying activity over long time can be difficult. Information on the ability of nitrifying sludge to regain its nitrifying activity after long-term storage is still scarce. In this work, the physiological and kinetic changes during the reactivation and stabilization of a nitrifying sludge previously exposed to ampicillin (AMP) were evaluated in a sequential batch reactor (SBR) after its long-term storage (1 year) at 4 °C. After storage, both ammonium and nitrite oxidizing processes were slow, being nitrite oxidation the most affected step. During the reactivation stage (cycles 1-6), physiological and kinetic activity of the nitrifying sludge improved through the operating cycles, in both its ammonium oxidizing and nitrite oxidizing processes. At the end of the reactivation stage, complete nitrifying activity was achieved in 10 h, reaching ammonium consumption efficiencies (ENH4 +) close to 100% and nitrate yields (YNO3 -) of 0.98 mg NO3 --N/mg NH4 +-N consumed without nitrite accumulation. During the stabilization stage (cycles 7-17), results indicated that the sludge could maintain a steady-state respiratory process with restoration percentages of 100% for nitrifying specific rates (qNH4 + and qNO3 -) with respect to their values obtained before storage. Furthermore, during the addition of 15 mg AMP/L (cycles 18-21), the sludge preserved its metabolic capacity to biodegrade 90% of AMP in 2 h. Therefore, long-term storage of nitrifying sludge could be used to preserve nitrifying inocula as bioseeds for bioremediation and bioaugmentation strategies.

Cometabolic biotransformation of benzotriazole in nitrifying batch cultures.

Benzotriazole (BT) is a corrosion inhibitor widely distributed in aquatic environments. Little is known about the cometabolic capacity of stabilized nitrifying sludge to biotransform BT. The contribution of the nitrification process in the simultaneous oxidation of ammonium and biotransformation of BT (5 mg/L) was evaluated in 49 d batch cultures inoculated with a sludge produced in steady-state nitrification. The nitrifying sludge could consume BT in the obligate presence of ammonium. A higher cometabolic biotransformation capacity was obtained by increasing the initial ammonium concentration (100-300 mg N/L), reaching 2.3- and 5.8-fold increases for efficiency and specific rate of BT removal. At 300 mg NH4+-N/L, the sludge biotransform 40.8% of BT and 77.6% of ammonium which was completely oxidized into nitrate. In assays with allylthiourea added as specific inhibitor of ammonium monooxygenase (AMO), it was shown that the totality of BT cometabolic biotransformation was associated with the AMO activity. The addition of acetate did not favor heterotrophic biotransformation of BT. BT provoked inhibitory effects on nitrification. This is the first study showing the role of ammonium oxidizing bacteria in the cometabolic biotransformation of BT and their potential use for cometabolism application in treatment of wastewater contaminated with ammonium and BT.

Biological ammonium and sulfide oxidation in a nitrifying sequencing batch reactor: Kinetic and microbial population dynamics assessments.

A kinetic study was carried out in a sequencing batch reactor (SBR) (125 mg NH4+-N/L) inoculated with a physiologically stable nitrifying sludge not previously acclimated to sulfur compounds and fed at different initial sulfide concentrations (2.5-20.0 mg HS--S/L). Up to 10.0 mg HS--S/L, the nitrifying process kept stable and complete, reaching an ammonium consumption efficiency (ENH4+) of 100% and a nitrate yield (YNO3-) of 0.95 ± 0.03 mg NO3--N/mg NH4+-N consumed. At 15.0 and 20.0 mg HS--S/L, after an initial alteration in the nitrite oxidizing process, the YNO2- was decreasing throughout the cycles and the YNO3- increasing, obtaining in the last cycle at 20.0 mg HS--S/L, an ENH4+ of 100%, a YNO2- of zero, and a YNO3- of 0.80 mg NO3--N/mg NH4+-N consumed. At the end of the period at 20.0 mg HS--S/L, the specific rates of ammonium consumption and nitrate formation were 15 and 55% lower than their respective values in the control period without sulfide addition, showing that the sludge had a better metabolic adaptation for ammonium oxidizing activity than for nitrite oxidizing activity. The sludge acquired a higher sulfide oxidation capacity along the cycles. Bacterial population dynamics assessment indicated that the ammonium oxidizing bacteria (AOB) community was more diverse and stable than the nitrite oxidizing bacteria (NOB) community. The use of consortia with a previously stabilized nitrifying activity in SBR may constitute an alternative for eliminating simultaneously ammonium by nitrification and sulfide by sulfide oxidation and be implemented for the treatment of wastewater with ammonium and sulfide.

Ampicillin biotransformation by a nitrifying consortium.

Ampicillin is a widely used ß-lactam antibiotic that has been detected in various effluents and can alter biological processes used in wastewater treatment such as nitrification. Physiological and kinetic behaviour of a nitrifying consortium in the presence of ampicillin (AMP) (10, 25, and 50 mg/L) was evaluated in batch cultures. Under the experimental conditions (320 ± 8 mg bacterial protein/L, C/N = 2.4, 24 h), the nitrifying behaviour was very similar among the controls without AMP and the assays with antibiotic, as there was no AMP effect on efficiency (ENH4+ = 99.7 ± 4.2%), yields (YNO2- = 0, YNO3- = 1.0 ± 0.1 mg N/mg NH4+-N consumed), neither specific rates of NH4+ oxidation and NO3- formation. Therefore, nitrifying bacteria were insensitive to AMP presence. At all assayed concentrations, after 24 h, 70.5 ± 3.7% of AMP was removed from the cultures through abiotic (16.0-16.5%), biosorption (23.2-47.0%), and biotransformation (10.0-29.8%) processes. With the increase in the initial AMP concentration, a greater participation of the biotransformation process, associated to an increase in the specific AMP consumption rate was attained. The sludge was able to completely oxidize NH4+ to NO3- by nitrification and eliminate AMP biologically, but without reaching its full mineralization.

Physiologic impact of 2-chlorophenol on denitrification process in mixture with different electron sources.

The aim of this study was to evaluate the physiologic behavior of sludge in the absence and presence of 2-chlorophenol (2-CP) with different electron donors (phenol, glucose, and acetate) during denitrification process. In batch assays with phenol in the presence of 2-CP, a significant decrease of phenol consumption efficiencies (E phenol) up to 99% was observed regarding the cultures without 2-CP. However, in most of the cases, nitrate consumption efficiencies ( E NO 3 - ), and yields of nitrogen gas ( Y N 2 ) and bicarbonate ( Y HCO 3 - ) were high, showing that the denitrifying respiratory process successfully occurred with phenol and 2-CP. The specific consumption rates of nitrate ( q NO 3 - ) and phenol (q phenol) decreased up to 6.0 and 32.3 times, respectively. In assays with glucose in the presence of 2-CP, the denitrifying performance was not significantly altered in terms of efficiencies and product yields; however, q NO 3 - was up to 1.6 times smaller than that obtained without 2-CP whereas q glucose was increased up to 1.17 times. In assays with acetate plus 2-CP, the E NO 3 - , E acetate, and Y N 2 values remained high but 2-CP caused a decrease in Y HCO 3 - . Moreover, q NO 3 - and q acetate increased up to 1.4 and 2.0 times, respectively. These results show that the negative or positive effects of 2-CP on denitrification process depend on the type and concentration of electron source. The obtained physiologic and kinetic information might be useful to define strategies to maintain successful denitrification processes in wastewater treatment bioreactors fed with 2-CP.

Implications of electric potentials applied on a denitrifying process.

The effect of three electric potentials (EP) (+104, -187 and -279 mV) applied to the denitrifying process was explored. It was observed that the denitrifying sludge was able to support the oxidation of p-cresol with the application of the EP in the absence of nitrate, but it was unable to drive the denitrification without an organic electron donor. On denitrification, the applied EP uncoupled the oxidative from the reductive process, favoring the p-cresol oxidation over the production of N2. Additionally, biochemical level effects were observed. At +104 and -279 mV potentials, the nitrate and nitrite consumption was affected as well as the p-hydroxybenzoate transformation. However, at -187 mV, effects seemed to occur only on the transport of substrates. This paper presents evidence that denitrification has very characteristic and different physiological behaviors for each EP assayed.

Inhibitory effects of quinoid redox mediators on a denitrifying culture.

Denitrification with p-cresol as the electron source was studied in the presence of three quinones at different molar concentrations (0-2 mM): menadione (MEN), alizarine (ALZ) and anthraquinone-2,6-disulfonate (AQDS). Results showed that denitrifying yields were not altered (0.9), but the substrates' consumption efficiency was mainly affected when adding MEN. In the presence of ALZ and MEN, specific consumption rates decreased 40% for p-cresol and 90% for nitrate. The sludge previously exposed to quinones was submitted to recovering denitrifying studies using acetate and p-cresol. After exposing to AQDS and ALZ, the metabolic activity of denitrifying sludge was completely recovered. The previous exposition to any MEN concentration resulted in a very low metabolic recuperation. The results show that MEN and ALZ have a marked inhibitory effect on substrates' consumption and the AQDS does not affect at all. The evidence suggests that MEN modifies the transport system of substrates and ALZ forms a complex with molybdenum. A model based on oxido-reduction potentials of the enzymes involved points out that the influence of quinones tested appears to be more associated with quinone moiety structural properties and hydrophobicity.

Decrease of inhibitory effect of 2-chlorophenol on nitrification in sequencing batch reactors.

The metabolic and kinetic behaviour of a nitrification process in the presence of 2-chlorophenol (2-CP) was evaluated in two sequencing batch reactors (SBR1, SBR2) inoculated with nitrifying sludge previously exposed to phenolic compounds. The SBR1 was inoculated with sludge previously exposed to 2-CP, while the SBR2 was inoculated with sludge previously exposed to p-cresol. An inhibitory effect of 20 mg 2-CP-C/L on both nitrification processes was observed, as specific rates decreased according to a control assay in the absence of 2-CP. However, the inhibitory effect decreased throughout the cycles. At the end of cycle 6, a stable nitrifying process was observed with the sludge previously exposed to 2-CP (SBR1), as an ammonium consumption efficiency and a nitrate production yield close to 99.6 ± 0.3% and 0.99 ± 0.02 were respectively achieved. Despite a complete ammonium consumption being achieved with the sludge previously exposed to p-cresol (SBR2), partial nitrification was observed as nitrate production yield accounted for 0.28 ± 0.08 and nitrite was accumulated within the culture. Nevertheless, both nitrifying sludges had the ability to completely consume 2-CP. The use of SBR systems with nitrifying sludge previously exposed to 2-CP resulted in a better nitrification performance, thus it may be a good alternative for achieving a stable nitrifying respiratory process where complete and simultaneous ammonium and 2-CP consumption can be acquired.

2-Chlorophenol consumption by cometabolism in nitrifying SBR reactors.

Cometabolic consumption of 2-chlorophenol (2-CP) by a nitrifying sludge was evaluated in two SBR reactors fed with 60 mg 2-CP-C/L and different initial ammonium concentrations (100, 200, 300, 400, and 500 mg NH4+-N/L). Irrespectively to the increase in ammonium concentration and throughout the operational cycles, the sludge achieved a complete nitrification in 14 days, accounting for ammonium consumption efficiencies close to 99% and nitrate production yields between 0.93 and 0.99. The sludge was able to completely consume 2-CP within 7 days. The increase in ammonium concentration provoked an increment in the specific rates of both ammonium (qNH4+-N) and 2-CP (q2-CP-C) consumption up to 5.2 and 3.1 times, respectively. The cometabolic effect of the increase in ammonium concentration on 2-CP consumption was supported by a direct and significant relationship between the qNH4+-N and q2-CP-C (r = 0.83). Moreover, batch assays conducted with ammonium, 2-CP, allylthiourea as specific inhibitor of the ammonium monooxygenase (AMO) enzyme, and the sludge inoculated into the reactors, resulted in a decrease of 34% in q2-CP-C, evidencing the participation of the AMO in the consumption of 2-CP. When the same assays were carried out with the sludge obtained from the SBR reactors after 13 operating cycles, a higher participation of the AMO in 2-CP consumption was noticed with a decrease of 53% in q2-CP-C. According to these results, the use of nitrifying sludge and high ammonium concentrations in SBR systems can be a suitable alternative for increasing the cometabolic consumption of recalcitrant compounds like 2-CP.

Simultaneous oxidation of ammonium and cresol isomers in a sequencing batch reactor: physiological and kinetic study.

The aim of this study was to evaluate the physiological and kinetic capacities of a nitrifying consortium to simultaneously oxidize ammonium (138 mg N/L day), m-cresol, o-cresol, and p-cresol (180 mg C/L day in mixture) in a sequencing batch reactor (SBR). A 1-L SBR was firstly operated without cresol addition (phase I) for stabilizing the nitrification respiratory process with ammonium consumption efficiencies close to 100 % and obtaining nitrate as the main end product. When cresols were added (phase II m-cresol (10, 20, and 30 mg C/L); phase III m-cresol (30 mg C/L) and o-cresol (10, 20, and 30 mg C/L); phase IV a mixture of three isomers (30 mg C/L each one)), inhibitory effects were evidenced by decreased values of the specific rates of nitrification compared with values from phase I. However, the inhibition diminished throughout the operation cycles, and the overall nitrifying physiological activity of the sludge was not altered in terms of efficiency and nitrate yield. The different cresols were totally consumed, being o-cresol the most recalcitrant. The use of SBR allowed a metabolic adaptation of the consortium to oxidize the cresols as the specific rates of consumption increased throughout the cycles, showing that this type of reactor can be a good alternative for treating industrial effluents in a unique reactor.

Mineralization of 2-chlorophenol by sequential electrochemical reductive dechlorination and biological processes.

In this work, a novel approach was applied to obtain the mineralization of 2-chlorophenol (2-CP) in an electrochemical-biological combined system where an electrocatalytic dehydrogenation process (reductive dechlorination) was coupled to a biological denitrification process. Reductive dechlorination of 2-CP was conducted in an ECCOCEL-type reactor on a Pd-Ni/Ti electrode at a potential of -0.40V vs Ag/AgCl(s)/KCl(sat), achieving 100 percent transformation of 2-CP into phenol. The electrochemically pretreated effluent was fed to a rotating cylinder denitrifying bioreactor where the totality of phenol was mineralized by denitrification, obtaining CO2 and N2 as the end products. The total time required for 2-CP mineralization in the combined electrochemical-biological process was 7.5h. This value is close to those previously reported for electrochemical and advanced oxidation processes but in this case, an efficient process was obtained without accumulation of by-products or generation of excessive energy costs due to the selective electrochemical pretreatment. This study showed that the use of electrochemical reductive pretreatment combined with biological processes could be a promising technology for the removal of recalcitrant molecules, such as chlorophenols, from wastewaters by more efficient, rapid, and environmentally friendly processes.

Kinetic Constants for Biological Ammonium and Nitrite Oxidation Processes Under Sulfide Inhibition.

Inhibition of nitrification by sulfide was assessed using sludge obtained from a steady-state nitrifying reactor. Independent batch activity assays were performed with ammonium and nitrite as substrate, in order to discriminate the effect of sulfide on ammonium and nitrite oxidation. In the absence of sulfide, substrate affinity constants (K S,NH4 = 2.41 ± 0.11 mg N/L; K s, NO2 = 0.74 ± 0.03 mg N/L) and maximum specific rates (q max,NH4 = 0.086 ± 0.008 mg N/mg microbial protein h; q max,NO2 = 0.124 ± 0.001 mg N/mg microbial protein h) were determined. Inhibition of ammonium oxidation was no-competitive (inhibition constant (K i , NH4 ) of 2.54 ± 0.12 mg HS(-)-S/L) while inhibition of nitrite oxidation was mixed (competitive inhibition constant (K' i , NO2 ) of 0.22 ± 0.03 mg HS(-)-S/L and no-competitive inhibition constant (K i , NO2 ) of 1.03 ± 0.06 mg HS(-)-S/L). Sulfide has greater inhibitory effect on nitrite oxidation than ammonium oxidation, and its presence in nitrification systems should be avoided to prevent accumulation of nitrite. By simulating the effect of sulfide addition in a continuous nitrifying reactor under steady-state operation, it was shown that the maximum sulfide concentration that the sludge can tolerate without affecting the ammonium consumption efficiency and nitrate yield is 1 mg HS(-)-S/L.

p-Cresol mineralization and bacterial population dynamics in a nitrifying sequential batch reactor.

The ability of a nitrifying sludge to oxidize p-cresol was evaluated in a sequential batch reactor (SBR). p-Cresol was first transformed to p-hydroxybenzaldehyde and p-hydroxybenzoate, which were later mineralized. The specific rates of p-cresol consumption increased throughout the cycles. The bacterial population dynamics were monitored by using denaturing gradient gel electrophoresis (DGGE) and sequencing of DGGE fragments. The ability of the sludge to consume p-cresol and intermediates might be related to the presence of species such as Variovorax paradoxus and Thauera mechernichensis. p-Cresol (25 to 200mgC/L) did not affect the nitrifying SBR performance (ammonium consumption efficiency and nitrate production yield were close to 100% and 1, respectively). This may be related to the high stability observed in the nitrifying communities. It was shown that a nitrifying SBR may be a good alternative to eliminate simultaneously ammonium and p-cresol, maintaining stable the respiratory process as the bacterial community.

Consumption of 2-chlorophenol using anaerobic sludge: physiological and kinetic analysis.

Chlorophenols are toxic and recalcitrant compounds produced by many industrial. Different strategies have been used to improve their biological consumption, but there is insufficient information to understand how the process is carried out. The objective of this study was to evaluate in batch tests the effect of the addition of phenol, acetate, or glucose as electron donors at different concentrations on the efficiencies, yields, and specific rates of 2-chlorophenol (2-CP) consumption. The addition of phenol (177.6 mg C/L), acetate (127.6 mg C/L), or glucose (77.6 mg C/L) increased the 2-CP consumption efficiency up to 54.6, 98.6, and 97.8 %, respectively. With respect to the control assay without electron donor, the specific rate of 2-CP consumption was up to 2.5 times higher with phenol (177.6 mg C/L), 8.4 times higher with acetate (127.6 mg C/L), and 3 times higher with glucose (127.6 mg C/L). The results showed that the type and concentration of electron donor determine the physiological behavior of the anaerobic sludge, modifying efficiency, yield, and specific rate values of the 2-CP consumption process. The addition of readily oxidable cosubstrates seems to be a good alternative and might be used for the biological treatment of industrial wastewater polluted with chlorinated phenols.

Effect of phenol and acetate addition on 2-chlorophenol consumption by a denitrifying sludge.

Chlorophenols are widely distributed in the environment. Various strategies have been used to improve their biological elimination under anaerobic conditions; however, such information is still scarce. The aim of this study was to evaluate in batch assays the consumption of 2-chlorophenol (2-CP) by a denitrifying sludge and the influence of acetate or phenol as co-substrates in the 2-CP consumption. It was observed that phenol (69 and 92 mg phenol-C L(-1)) and acetate (60 and 108 mg acetate-C L(-1)) enhanced 2-CP consumption by the denitrifying sludge, increasing both the efficiency (up to 100%) and specific rate of 2-CP consumption. When phenol was added at 92 mg C L(-1), the specific consumption rate of 2-CP increased 2.6 times with respect to the control lacking co-substrates, whereas with acetate (108 mgC L(-1)) the increase was 9.0 times. Acetate appeared to be a better co-substrate for 2-CP consumption, obtaining a specific consumption rate of 2.48 +/- 0.14 mg 2-CP-C g(-1) VSS d(-1) at 108 mg acetate-C L(-1). The mass balance analysis indicated that the denitrifying sludge was able to simultaneously mineralize 2-CP, phenol or acetate (E2-CP, E(Phenol), and E(Acetate) close to 100% [E = consumption efficiency], Y(HCO3-) of 0.90 +/- 0.10 [Y = yield]) and reduce nitrate to nitrogen gas (E(NO3-) of 100% and Y(N2) of 0.96 +/- 0.02). It was shown that the addition of co-substrates as phenol or acetate could be a good alternative for improving the elimination of chlorophenols from wastewaters by denitrifying sludges.

2-Chlorophenol consumption and its effect on the nitrifying sludge.

The kinetic behavior of a nitrifying sludge exposed to 2-chlorophenol (2-CP) was evaluated in batch culture. The assays were performed using a stabilized nitrifying sludge. In control assays with (mg L(-1)): NH(4)(+)-N (100) and NaHCO(3)(-)-C (250), the substrates were consumed in 8h, the ammonium consumption efficiency was 99% and the NO(3)(-) yield higher than 0.9. When 5mg 2-CP-C L(-1) was added, it was transformed into an unidentified intermediate and the nitrifying efficiency decreased to 10%. Ammonium specific consumption rate diminished 95%, but the NO(3)(-) yield remained higher than 0.9. The biomass previously exposed to 2-CP was newly suspended with NH(4)(+)-N or NO(2)(-)-N in order to evaluate the ammonium and nitrite oxidizing processes. The consumption efficiencies and NO(3)(-) yields were similar to those obtained in control assays. However, the total time required for ammonium and nitrite consumption increased to 120 and 42 h, respectively. Specific consumption rates for NH(4)(+)-N and NO(2)(-)-N decreased by 95% and 83% respectively, compared to control assays. Thus, the previous contact to 2-CP had more influence on ammonium oxidizing process than the nitrite oxidizing process. These are the first evidences where a nitrifying sludge exposed to 2-CP are reported.

Kinetic limitations during the simultaneous removal of p-cresol and sulfide in a denitrifying process.

The aim of this study was to evaluate the capacity of a denitrifying consortium to achieve the simultaneous removal of nitrate, sulfide and p-cresol and elucidate the rate-limiting steps in the mixotrophic process. Nitrite reduction appeared as the most evident rate-limiting step in the denitrifying respiratory process. The nitrite reduction rate achieved was up to 57 times lower than the nitrate reduction rate during the simultaneous removal of sulfide and p-cresol. Negligible accumulation of N(2)O occurred in the denitrifying cultures corroborating that nitrite reduction was the main rate-limiting step of the respiratory process. A synergistic effect of nitrate and sulfide is proposed to explain the accumulation of nitrite. The study also points at the oxidation of S(0) as another rate-limiting step in the denitrifying process. Different respiratory rates were achieved with the distinct electron donors provided (p-cresol and sulfide). The oxidation rate of p-cresol (q(CRES)) was generally higher (up to 2.6-fold in terms of reducing equivalents) than the sulfide oxidation rate (q(S2-)), except for the experiments performed at 100 mg S(2-) L(-1) in which q(S2-) was slightly (approximately 1.4-fold in terms of reducing equivalents) higher than q(CRES). The present study provides kinetic information, which should be considered when designing and operating denitrifying reactors to treat industrial wastewaters containing large amounts of sulfurous, nitrogenous and phenolic contaminants such as those generated from petrochemical refineries.

Effects of different quinoid redox mediators on the simultaneous removal of p-cresol and sulphide in a denitrifying process.

The catalytic effects of different quinoid redox mediators (RM) on the simultaneous removal of sulphide and p-cresol in a denitrifying process were evaluated in batch studies. 2-Hydroxy-1,4-naphthoquinone (LAW) and anthraquinone-2,6-disulphonate (AQDS) did not significantly affect the sulphide oxidation rate, which, in contrast, was increased 14% in the presence of 1,2-naphthoquinone-4-sulphonate (NQS). The input of NQS on the oxidation of sulphide was also favourably reflected in a 13% higher sulphate production. All RM promoted a higher (up to 34% compared to the control lacking RM) degree of mineralization of p-cresol. LAW also supported a 47% higher denitrifying yield (Y(N2)), compared to the control lacking quinones. Nevertheless, AQDS and NQS decreased the Y(N2) by 12-13%. Our results suggest that a proper scrutiny should be conducted before deciding the sort of quinone to be applied in denitrifying processes. The heterogeneous effects observed also advise to consider both the respiratory rates and the yields as important parameters for deciphering the impact of RM on denitrifying processes.

Effect of initial sulfide concentration on sulfide and phenol oxidation under denitrifying conditions.

The objective of this work was to evaluate the effect of the initial sulfide concentration on the kinetics and metabolism of phenol and sulfide in batch bioassays using nitrate as electron acceptor. Complete oxidation of sulfide (20 mg L(-1) of S(2-)) and phenol (19.6 mg L(-1)) was linked to nitrate reduction when nitrate was supplemented at stoichiometric concentrations. At 32 mg L(-1) of sulfide, oxidation of sulfide and phenol by the organo-lithoautotrophic microbial culture was sequential; first sulfide was rapidly oxidized to elemental sulfur and afterwards to sulfate; phenol oxidation started once sulfate production reached a maximum. When the initial sulfide concentration was increased from 20 to 26 and finally to 32 mg L(-1), sulfide oxidation was inhibited. In contrast phenol consumption by the denitrifying culture was not affected. These results indicated that sulfide affected strongly the sulfide oxidation rate and nitrate reduction.