The performance of Biostyr media using an upflow biofilter scheme was investigated. Three separate Modes that varied the organic strength of the influent at an operating temperature of 12°C were evaluated to explore how nitrification and denitrification responded to different concentrations of biodegradable COD loadings. Effluent suspended solids and observed yield coefficients were also analyzed along with ways to determine kinetic parameters. Combinations of primary and secondary effluent were used as the influent using domestic wastewater from a primary sewer of the Blacksburg VPI Sanitation Authority, in Blacksburg, Virginia, located on the Virginia Tech Campus. Reduction of nitrification rates was observed in the aerobic reactor at biodegradable COD loadings to the aerobic biofilter of 2.2 kg/day/m³ and greater. Complete denitrification was observed in the anoxic tank at biodegradable COD loadings to the anoxic biofilter of 7.5 kg/day/m³ and greater. Above this value, denitrification was limited by the available amount of NOx-N. An optimum influent biodegradable COD loading rate of 7.5 kg/day/m³ to the anoxic biofilter and 2.2 kg/day/m³ to the aerobic biofilter was experimentally demonstrated for simultaneous nitrification and denitrification. When secondary effluent was fed to the reactors, complete nitrification was observed indicating success in terms of ammonia polishing. However, low concentrations of influent biodegradable COD limited the ability for the anoxic reactor to denitrify. Thus, it was determined that the addition of an external carbon source would need to be implemented to denitrify when using low strength secondary effluent. Effluent total suspended solids were consistently below 20 mg/L indicating that secondary clarification would not be needed for this reactor scheme. The combined observed yield was consistently around 0.2 (mg-VSS/mg-COD consumed) or less. However, the evaluation of separate anoxic and aerobic yields was inconsistent when averaging data over 5 days, because of the recycle of biomass from the aerobic reactor back to the anoxic reactor. The kinetic parameters, q'_{MAX NIT} and K’_{S NIT} were calculated by plotting nitrification rates as a function of ammonia concentration. These coefficients were calculated by using separate segments within the media under less than optimal conditions. It was determined that analyzing the whole system rather than segments would be better suited for kinetic parameter analysis. However, there was not enough collected data to achieve this. Thus, ranges identifying further influent biodegradable COD loadings were recommended for later research to better a complete kinetic parameter evaluation. / Master of Science

Sanafoam Vaporooter II is a trademark name for a process which uses metham sodium and dichlobenil to remove tree roots from municipal sewer and storm drain lines. One or more of the chemicals in this process, or their degradation products, have been suspected of disrupting the nitrification process in waste treatment plants downstream of the points of application. This work was undertaken to identify the chemical responsible for the disruption, and to recommend means for assuaging the problem. The impacts of the herbicides used in the Vaporooter II process were separately tested in bench scale reactors. Metham sodium was found to be the herbicide responsible for disrupting nitrification. It did so at concentrations of 4 mg/L or higher in systems which have a mixed liquor volatile suspended solids concentrations of 1800 mg/L. This corresponds to a herbicide to biomass ratio of 0.017 moles/gram. The stability of metham sodium was also tested in bench scale experiments. The compound was found to be fairly stable at the pH values, temperatures and oxygen concentrations typically found in sewage collection systems and so undergoes little degradation after application. Powdered activated carbon (PAC) is effective in absorbing metham sodium from raw sewage. In bench scale reactors, application rates of 10 mg PAC per 1 mg metham sodium reduced metham sodium concentrations to levels which did not disrupt the nitrification process. / Master of Science

Interest in transport of dissolved nitrogen (N) and carbon (C) in forested ecosystems is growing because of potential effects of these solutes on streamwater quality and implications for C sequestration. Additional research will further the understanding about the dynamics of these soil solutes, particularly in response to harvesting of forests. Also, the purported role of riparian buffers, where logging is restricted along stream channels, in retaining soil solutes is not well studied in the steeply sloping terrain of the southern Appalachian Mountains. I examined solute transport in a first-order watershed in the Nantahala National Forest in North Carolina that was harvested in February 2006 with retention of a 10-m riparian buffer. To quantify the movement of dissolved inorganic nitrogen (DIN), dissolved organic nitrogen (DON), and dissolved organic carbon (DOC), four transects of lysimeters, approximately 30 m apart, were installed perpendicular to the stream on one hillslope. Porous ceramic cup (2-bar) lysimeters were installed in each transect 1, 4, 10, 16, 30, and 50 m from the stream in the A horizon and B horizon, and 4, 16, and 50 m from the stream in the saprolite layer. Samples were removed from the lysimeters 24 hr after 50 centibars of tension were placed on them, and riparian groundwater well and stream samples were collected at the same time as lysimeter samples. Collection of samples from the lysimeters, wells, and stream occurred every four to six weeks for one calendar year beginning March 2007. A 16-wk laboratory N mineralization study was conducted on A horizon soils. Mean nitrate values in the soil solution of the A horizon in the spring were 1.53mg-N/L and decreased through the growing season to 0.030mg-N/L. Mean soil solution nitrate values in the B horizon and saprolite layer were 0.40mg-N/L in the spring and summer and decreased to 0.031mg-N/L in the winter. Mean soil solution ammonium concentrations were higher in the A horizon (0.090mg-N/L) than the B horizon and saprolite layer (0.034mg-N/L) and were lowest during the summer and fall. Dissolved organic C was significantly higher in the A horizon, with values ranging from 2.3mg/L to 599mg/L, than in the relatively stable B horizon and saprolite (1.9mg/L to 36.6mg/L). Dissolved organic C was logarithmically correlated to DON (r2 = 0.64), and DON values were highest in the A horizon (0.70mg/L). Cumulative N mineralization potential ranged from 48.1mg-N/kg to 75.6mg-N/kg and was not a useful predictor for nitrate soil solution values. Nitrate leached vertically, and a large percentage of nitrate was stored in the B horizon and saprolite. Ammonium, DON, and DOC did not appear to leach vertically because they did not increase in the B horizon or saprolite layer. Ammonium, DON, and DOC are less mobile in soil solution than nitrate. The 10-m riparian zone had little impact on nitrate, ammonium, DON, and DOC removal. Nitrate remaining in the A horizon was likely removed through plant uptake in the harvested area before reaching the riparian zone. There was no detectable difference between ammonium concentrations in the harvested area and riparian zone likely because of limited mobility. The riparian zone did not remove excess DON or DOC, and in some transects was a source of DON and DOC. Nitrate and DOC concentrations were highly variable among transects and locations within transects. This may be caused by sensitivity of these solutes to site heterogeneity. This suggests that a large number of lysimeters should be used to account for this variability in future studies to ensure accuracy. This study observed limited vertical leaching of ammonium, DON, and DOC through the profile. However, excess nitrate was observed moving from the A horizon into the B horizon and saprolite layer, suggesting the potential for delivery to the stream via subsurface transport and the need for attenuation of nitrate by the riparian zone. Because of low concentrations of nitrate entering the riparian zone during this study, the capacity for riparian attenuation of nitrate was not demonstrated. / Master of Science

The purpose of this research was to characterize a flushed dairy manure wastewater and to develop the kinetic and stoichiometric parameters associated with nitrogen removal from the wastewater, as well as to demonstrate experimental and simulated nitrogen removal from the wastewater. The characterization showed that all the wastewaters had carbon to nitrogen ratios large enough for biological nitrogen removal. Analysis of carbon to phosphorus ratios showed that enough carbon is available for phosphorus removal but enough may not be available for both nitrogen and phosphorous removal in anaerobically pretreated wastewater. In addition, kinetic and stoichiometric parameters were determined for the biological nitrogen removal in sequencing batch reactors for the dairy manure wastewater. Results showed that many parameters are similar to those of municipal wastewater treatment systems. This characterization and the derived kinetic and stoichiometric parameters provided some of the information necessary for development of a nitrogen removal process in a sequencing batch reactor. Lab scale treatment of a 1:2 dilution of the anaerobically pretreated wastewater was demonstrated. Treatment was able to achieve between 89 and 93% removal of soluble inorganic nitrogen as well as up to 98% removal of biodegradable soluble and colloidal COD. In addition, a solids removal efficiency of between 79 and 94% was achieved. The lab scale treatment study demonstrated that sequencing batch reactors are capable of achieving high nitrogen removal on wastewaters with the carbon to nitrogen ratios of the dairy manure wastewater. Model simulations of the treatment process were used to develop a sensitivity analysis of the reactor feed configuration as well as the kinetic and stoichiometric parameters. The analysis of the feed configuration demonstrated the advantage of decreasing the amount of feed that is fed in the last feed period so that the effluent nitrate will be minimized. The analysis indicated that the autotrophic growth rate is one of the most important parameters to measure while error in the heterotrophic decay or yield values can lead to miscalculations of oxygen required for treatment. / Master of Science

Metalaxyl, [N—(2,6-Dimethylphenyl)-N-(Methoxyacetyl)-alanine methyl ester], is used extensively in tobacco (Nicotiana tabacum L.) production for prevention of black shank (Phytophthora parasitica Dast. var. nicotianae), blue mold (Peronospora tabacina Adam), and damping-off (Pythigm spp.). Metalaxyl is also patented as a nitrification inhibitor, although not marketed for that purpose. Proper maturity and ripening of flue-cured tobacco depends on an adequate supply of N through the time of removal of the inflorescence, with a declining supply of N from that point. Use of a chemical which might prolong the availability of N in tobacco could delay maturity and reduce the quality of the cured leaf. These studies were conducted to determine whether metalaxyl might inhibit nitrification under a broad range of soil physical and environmental conditions prevalent in the tobacco producing areas of Virginia. The influence of soil type, soil pH, soil temperature, and soil moisture on inhibition of nitrification by metalaxyl (1 mg kg⁻¹) were investigated in three soils used extensively for tobacco production. Soils used in the study were Cecil sandy loam (clayey, kaolinitic, thermic Typic Hapludult), Appomattox fine sandy loam (clayey, mixed, thermic Typic Kandhapludult), and Mattoponi sandy loam (clayey, mixed, thermic Typic Hapludult). Metalaxyl did not inhibit nitrification under any of the conditions studied. However, NO₂⁻ accumulation with metalaxyl was sometimes greater than the control, especially at high pH (7.0) in the Cecil and Appomattox soils, and at 10 and 20°C. Nitrite and NO₃⁻ accumulations from four rates of metalaxyl (1, 5, 25, and 125 mg kg⁻¹) were compared with those of an untreated control and a nitrapyrin standard over a seven week soil incubation period in further studies using the same soils and adjusted pH levels. Significant NO₂⁻ accumulation occurred during the first week after treatment at high pH in all soil types, with 5, 25, and 125 mg kg⁻¹ metalaxyl. Only the 125 mg kg⁻¹ metalaxyl treatment caused NO₂⁻ accumulation at the high pH in all soils beyond the second week after treatment, with the peak occurring in most cases between weeks three and four. Nitrate accumulation proceeded normally in all soil types and pH levels except with treatments of 25 and 125 mg kg". Nitrate accumulations with 25 mg kg⁻¹ were similar to those for nitrapyrin. The 125 mg kg⁻¹ rate was consistent in causing near total inhibition of NO₃⁻ accumulation at all pH levels in all soils. Nitrate accumulation tended to be lower at lower soil pH levels compared to the highest pH for all soils. Little difference in nitrification due to soil appears to be evident. Use of metalaxyl at recommended rates of 0.25 to 1.5 mg kg⁻¹ would not be expected to inhibit nitrification. / Ph. D.

Dans l'étang de Berre, les apports excessifs en eau douce entraînent souvent des variations brusques de la salinité et des apports en azote parfois abondants. Les processus de régénération sédimentaire et d'élimination d'azote constituent des phénomènes majeurs pouvant expliquer le développement phytoplanctoniques et contrôler l'eutrophisation. L'étude de ces processus a été réalisée dans 2 stations ayant subi des historiques de variations de salinité différents. RDNA et anammox étaient négligeables. La dénitrification (couplée à la nitrification) serait le principal processus de sortie de N2 de l'étang. La forte production primaire pélagique utilisant l'ammonium a pu être expliquée par les processus de régénération, contrairement au nitrate dont la demande pélagique serait soutenue par des apports externes. Comme la salinité est susceptible d'influencer les processus d'azote, des microcosmes ont été utilisés afin d'évaluer la sensibilité de ces processus (résistance, résilience, shift) face aux variations à court terme de la salinité. Les 2 stations ont présenté des réponses différentes (dénitrification et nitrate réduction plus résilientes que la nitrification), suggérant que les stress halins à long terme influencent les réponses à court terme. Une hypothèse serait que la diversité et la structure des communautés joueraient un rôle crucial dans le maintien des taux d'activité. Dans cette étude, seule la communauté nitrifiante (AO) a été étudiée. Les AO et leurs transcrits étaient spécifiques aux sites et leurs abondances changeaient faiblement entre les microcosmes (résistance plus forte dans le site fréquemment soumis à des fluctuations de salinité). / The Berre lagoon receives excess freshwater leading to strong changes in the ecosystem salinity and in the nutrient inputs. Regeneration processes and the N2 removal of nitrogen are of a particular interest as they can explain the planktonic development and control the eutrophication state. These processes were studied in 2 stations with different patterns of long term stresses. DNRA and anammox were negligible. Denitrification (particularly the one coupled to nitrification) would be the main N2 removal in the lagoon. The high primary production based on N-NH4+ can be explained by the regeneration processes, while the primary production based on N-NO3- might be sustained by external inputs. As salinity susceptible to influence directly or indirectly the nitrogen processes, a microcosm approach was used in order to assess their sensitivity (resistance, resilience, shifts) to short term salinity fluctuations. Overall, towards the same kind of perturbations, the two stations responded differently (denitrification and nitrate reduction exhibited showed higher resilience than nitrification.), suggesting that long-term saline stresses would influence short-term responses. We suggest that the diversity and the structure of the communities would be crucial in their functional redundancy and thereby in the maintenance of the rates. In this study, we focused on the total structure of the nitrifying community (AO). AO and their transcripts were site-specific and their abundances changed slightly between microcosms (higher resistance in the site subject to frequent salinity fluctuations compared to the other site which is not affected by changes in the in situ salinity).

There is an urgent need to develop ammonia removal treatment systems for municipal and industrial wastewater treatment due to the increasingly stringent ammonia effluent discharge regulations implemented by Canada, the United States, and the European Union. The objective of this dissertation is to develop new understanding and advance the current design and operation of total ammonia nitrogen (TAN) removal via the moving bed biofilm reactor technology (MBBR) for municipal and industrial wastewaters. The first specific objective is to develop a passive, low operationally intensive, efficient and robust design strategy for municipal wastewater treatment to achieve partial nitritation (PN) as a pre-treatment to anammox treatment without using control strategies such as operating at low dissolved oxygen, or the use of inhibitors. This first objective includes developing new knowledge of the biofilm, biomass and microbiome of attached growth PN systems. The second specific objective is to investigate the impact of defining a maximum biofilm thickness, via bio-carrier design, to enhance the effects of free nitrous acid inhibition for PN of municipal wastewaters. The third objective is to investigate the effect of influent copper concentration on nitrifying MBBR systems over long-term operations, to demonstrate the feasibility of the nitrifying MBBR as a solution for TAN removal from gold mining wastewaters. The results pertaining to the first objective, achieved via a study investigating the operation of a nitrifying moving bed biofilm reactor at elevated TAN surface area loading rates (SALRs) of 3, 4, 5, and 6.5 g TAN/m²∙d with the aim of achieving passive PN, demonstrates that operating at a TAN SALR value of 6.5 g TAN/m²∙d can achieve PN without restricting dissolved oxygen or using inhibitors. Operating at a TAN SALR value of 6.5 g TAN/m²∙d achieves a TAN surface area removal rate (SARR) of 3.5 g TAN/m²∙d, and a nitrite accumulation of 99.8% of the oxidized TAN, demonstrating the suppression of nitrite oxidizing bacteria (NOB) activity, while achieving elevated TAN SARR values. At the molecular-scale, there is a statistically significant change in the ammonia oxidizing bacteria (AOB) to NOB ratio from 1:2.6 to 8.7:1 as the TAN SALR increases from 3 to 6.5 g TAN/m²∙d; however, even at a TAN SALR value of 6.5 g TAN/m²∙d there is an NOB abundance of approximately 2%; thus demonstrating that NOB remain present in the biofilm, while their activity is suppressed by operation at elevated TAN SALR values. Furthermore, this system was shown to achieve stable PN consistently for over a period of 10 months of operation, demonstrating a robust, passive, low operational strategy for attached growth PN. The second objective of this dissertation is addressed through a study that compared the carrier design of defined maximal biofilm thickness (z-prototype carrier) to undefined maximal biofilm thickness (chip-prototype carrier) for PN via free nitrous acid inhibition of tertiary, low carbon, municipal wastewaters. The study demonstrates that defined maximal biofilm thickness is a preferred design choice to achieve attached growth PN. The chip-prototype carrier shows biofilm thicknesses and biofilm mass values that are ten-fold higher than the z-prototype carrier, which is shown to contribute to the impact of free nitrous acid on AOB and NOB activities. The z-prototype carrier shows PN is achieved after 3 hours of exposure to free nitrous acid while the chip-prototype carrier does not achieve PN within this same time of exposure. Therefore, the defined maximal biofilm thickness carrier is identified in this research as the preferred design option to achieve attached growth PN for municipal, low carbon, tertiary wastewater treatment. The results of the third objective, achieved via a study investigating the effects of influent copper concentrations on nitrifying MBBR during long term operations to gold mining wastewaters, demonstrates that there is no AOB inhibition in attached growth systems exposed to 0.1, 0.3, 0.45, and 0.6 mg Cu/L for long exposure times. A trend of increasing nitrite accumulation with increasing influent copper concentrations is shown, indicating that NOB inhibition occurs at influent copper concentrations of 0.3 mg Cu/L and greater, with the greatest NOB inhibition observed with an influent copper concentration of 0.6 mg/L. There is no statistically significant difference in biofilm characteristics at the copper concentrations tested; however, there is a trend of increasing biofilm thickness and biofilm roughness with increasing copper concentrations. This study demonstrates the resilience of the nitrifying biofilm to copper inhibition and demonstrates that the nitrifying MBBR is a promising system for removing TAN in mining wastewater in the presence of copper.

Nitrogen removal technologies can help dairy industries meet their nutrient management needs. This thesis investigated two separate treatment technologies for total ammonia nitrogen (TAN) removal: algae raceway ponds and aerated biofilm reactors. Six 1000- liter algae raceway ponds and four 1000-liter tote tanks, each equipped with 10 sheets of nonwoven geotextile (i.e., thermally bonded or needle-punched) biofilm substrate, were used to treat the effluent from a flush dairy in central coastal California (TAN = 251 mg/L, cBOD5 = 204 mg/L). For the algae raceway ponds (TAN loading rate = 7 - 35 g/m³-day among 7-, 10- and 14-day hydraulic residence times (HRT)), first-order removal rate constants (k) were ~0.2 day⁻¹ in the summer and 0.1 - 0.2 day⁻¹ in the winter. Removal rate constants had no correlations (R² < 0.1) with water temperature, weak to moderate (for 7-day ponds, R² = 0.55) correlations with insolation and weak to no correlations with biomass (i.e. volatile solids) concentration. During the winter, low insolation likely inhibited algal photosynthesis and biological TAN treatment. Ponds with 7-day HRT had distinct absence of nitrate and nitrite compared to 10- and 14-day ponds. Net productivities were ~20 g/m²-day in summer and 9 – 11 g/m²-day in winter; gross productivities were 120 – 160 g/m²-day in summer and 77 – 150 g/m²-day in winter. Productivities had no correlations (R² < 0.1) with water temperature and weak to moderate correlations (for 14-day ponds, net productivity R² = 0.56, gross productivity R² = 0.83) with insolation. Analysis of organic compounds in pond effluent showed dissolved volatile solids (~2300 mg/L) were mostly non-biodegradable (~98% of soluble oxygen demand). Dissolved organic nitrogen concentrations in the pond effluent were ~35 mg/L. For the aerated biofilm reactors, tanks with needle-punched geotextiles had greater first- order TAN removal rate constants (0.69 day⁻¹) than tanks with thermally bonded geotextiles (0.23 day⁻¹) while operating in batch mode. Needle-punched geotextile reactor also accumulated sludge faster and had higher attached to water column biomass (i.e. volatile solids) ratios (~0.08 g VS/g VS) than thermally bonded geotextile reactor (~0.04 g VS/g VS). Among the four tanks, mass of attached biomass was 150 – 340 g per tank while mass of biomass in the water column was 3290 – 5430 g per tank. Comparing the two treatment technologies, aerated biofilm reactors (removal = 64 – 77%, k = 0.2 – 0.3 day⁻¹, removal rate = 36 – 43 g-N/m²-day, 16 – 19 g-N/m³-day) had more removal and faster removal rates per square meter of land footprint compared to the algae raceway ponds (removal = 38 – 77%, k = 0.1 – 0.2 day⁻¹, removal rate = 4 – 5 g- N/m²-day, 13 – 17 g-N/m³-day), likely due to direct application of aerators in the reactors.

The influence of nitrification on the “soil” solution of container media has not been documented. The investigation of this influence is justified since the ionic form of N in a soil solution has a significant influence on plant tissue nutrient content and growth. Three genera of woody plants were grown in one-liter containers filled with pine bark, treated with and without a nitrification inhibitor and fertilized with 210 ml of a 100 ppm NH₄-N solution. Without the inhibitor and over time, “soil” solution NH₄-N concentrations and pH decreased and NO₃-N concentrations increased. “Soil” solution and tissue cation concentrations were generally greater without the inhibitor. In a second experiment, pine bark in one-liter containers was treated with either 0, 3 or 6 kg lime m⁻³. “Soil” solution data and NO₃-N accumulation rate (NAR) data showed an earlier nitrification of NH₄-N at the 6 kg lime compared to the 3 kg lime treatment whereas NO₃-N was not found at the 0 kg lime treatment. In a 3rd experiment, pine bark in one-liter containers was treated with 210 ml of either 25, 100 or 200 ppm NH₄-N. Over time “soil” solution NO₃-N concentrations were greatest and pH values were lowest at the 200 ppm N treatment. The NAR of the 25 ppm N treatment was less than the 100 and 200 ppm N treatment which were not different. The lack of correspondence between the “soil” solution NO₃-N data and the NAR data for the 100 and 200 ppm N treatments was explained on the basis of NH₄-N supply. In a 4th experiment, pine bark in one-liter containers were subjected to either 10°, 20°, 30° or 40° C for 24 days. “Soil” solution NH₄-N concentrations decreased over time at 10°, 20° and 30°. “Soil” solution NH₄-N and NO₃-N concentrations at 40° were considerably higher and lower, respectively, than at other temperatures. Over time the general order of NAR was: 20° = 30° > 10° > 40°. Results of these experiments indicate that nitrification is an important consideration in the nutrition of container-grown plants. / Ph. D.

Thesis (PhD (Chemical Engineering))--Cape Peninsula University of Technology, 2019 / Simultaneous nitrification and aerobic denitrification (SNaD) is a preferred method for single stage total nitrogen (TN) removal, which was recently proposed to improve wastewater treatment plant design. However, SNaD processes are prone to inhibition by toxicant loading with free cyanide (CN-) possessing the highest inhibitory effect on such processes, rendering these processes ineffective. Despite the best efforts of regulators to limit toxicant disposal into municipal wastewater sewage systems (MWSSs), free cyanide (CN-) still enters MWSSs through various pathways; hence, it has been suggested that CN- resistant or tolerant microorganisms be utilized for processes such as SNaD. To mitigate toxicant loading, organisms in SNaD have been observed to adopt a multiphase growth strategy to sequentially degrade CN- during primary growth and subsequently degrade TN during the secondary growth phase. However, CN- degrading microorganisms are not widely used for SNaD in MWSSs due to the inadequate application of suitable microorganisms (Chromobacterium violaceum, Pseudomonas aeruginosa, Thiobacillus denitrificans, Rhodospirillum palustris, Klebsiella pneumoniae, and Alcaligenes faecalis) commonly used in single-stage SNaD. The use of CN- degrading or resistant microorganisms for SNaD is a cost-effective method compared to the use of other methods of CN- removal prior to TN removal, as they involve multi-stage systems (as currently observed in MWSSs). The use of CN- degrading microorganisms, particularly when used as a consortium, presents a promising and sustainable resolution to mitigate inhibitory effects of CN- in SNaD. However, SNaD is known to be completely inhibited by CN- thus it is imperative to also study some thermodynamic parameters of SNaD under high CN- conditions to see the feasibility of the process. The Gibbs free energy is significant to understand the feasibility of SNaD, it is also vital to study Gibbs free energy to determine whether or not the biological reaction is plausible. The relationship between the rate of nitrification and Gibbs free energy was also investigated. The attained results showed that up to 37.55 mg CN-/L did not have an effect on SNaD. The consortia degraded CN- and achieved SNaD, with degradation efficiency of 92.9 and 97.7% while the degradation rate of 0.0234 and 0.139 mg/L/hr for ammonium-nitrogen (NH4-N) and CN- respectively. Moreover, all the free Gibbs energy was describing the individual processes were found to be negative, with the lowest Gibbs free energy being -756.4 and -1830.9 Kcal/mol for nitritation and nitratation in the first 48 h of the biological, reaction respectively. Additionally, a linear relationship between the rate of NH4-N and nitrite-nitrogen (NO2-N) degradation with their respective Gibbs free energy was observed. Linear model was also used to predict the relationship between NH4-N, NO2-N degradation and Gibbs free energy. These results obtained showed a good correlation between the models and the experimental data with correlation efficiency being 0.94 and 0.93 for nitritation, and nitratation, respectively. From the results found it can be deduced that SNaD is plausible under high cyanide conditions when cyanide degrading or tolerant microorganisms are employed. This can be a sustainable solution to SNaD inhibition by CN- compounds during wastewater treatment. Furthermore, a single strain was purified from the consortium and identified as Acinetobacter courvalinii. This bacterial strain was found to be able to perform sequential CN- degradation, and SNaD; an ability associated with multiphase growth strategy of the microorganism when provided with multiple nitrogenous sources, i.e. CN- and TN. The effect of CN- on nitrification and aerobic denitrification including enzyme expression, activity and protein functionality of Acinetobacter courvalinii was investigated. It was found that CN- concentration of up to 5.8 mg CN-/L did not affect the growth of Acinetobacter courvalinii. In cultures whereby the A. courvalinii isolate was used, degradation rates of CN- and NH4-N were found to be 2.2 mg CN-/L/h and 0.40 mg NH4-N/L/h, respectively. Moreover, the effect of CN- on NH4-N, nitrate-nitrogen (NO3-N) and NO2-N oxidizing enzymes was investigated, with findings indicating CN- did not affect the expression and activity of ammonia monooxygenase (AMO), but affected the activity of nitrate reductase (NaR) and nitrite reductase (NiR). Nevertheless, a slow decrease in NO2-N was observed after the addition of CN- thus confirming the activity of NaR and the activation of the denitrification pathway by the CN-. Moreover, five models’ (Monod, Moser, Rate law, Haldane, and Andrew’s model) ability to predict SNaD under CN- conditions, indicated that only Rate law, Haldane and Andrew’s models, were suited to predict both SNaD and CN- degradation. Due to low degradation rates of NH4-N and CN-, optimization of SNaD was essential. Therefore, response surface methodology was used to optimize the SNaD under CN- conditions. The physiological parameters that were considered for optimization were temperature and pH; with the result showing that the optimum for pH and temperature was 6.5 and 36.5oC respectively, with NH4-N and CN- degradation efficiency of 50 and 80.2%, respectively. Furthermore, the degradation kinetics of NH4-N and CN- were also studied under the optimum conditions in batch culture reactors, and the results showed that up to 70.6% and 97.3% of NH4-N and CN- were simultaneously degraded with degradation rates of 0.66 and 0.41 mg/L/h, respectively. The predictive ability of RSM was further compared with cybernetic models, and cybernetic models were found to better predict SNaD under CN- conditions. These results exhibited a promising solution in the management of inhibition effected of CN- towards SNaD at an industrial scale.