Biological breakdown of denitrifying sulfide removal process in high-rate expanded granular bed reactor

This work conducted a denitrifying sulfide removal (DSR) test in an expanded granular sludge bed (EGSB) reactor at sustainable loadings of 6.09 kg m⁻³ day⁻¹ for sulfide, 3.11 kg m⁻³ day⁻¹ for nitrate–nitrogen, and 3.27 kg m⁻¹ day⁻¹ for acetate–carbon with >93% efficiency, which is significantly higher than those reported in literature. Strains Pseudomonas sp., Nitrincola sp., and Azoarcus sp. very likely yield heterotrophs. Strains Thermothrix sp. and Sulfurovum sp. are the autotrophs required for the proposed high-rate EGSB-DSR system. The EGSB-DSR reactor experienced two biological breakdowns, one at loadings of 4.87, 2.13, and 1.82 kg m⁻³ day⁻¹; reactor function was restored by increasing nitrate and acetate loadings. Another breakdown occurred at loadings of up to 8.00, 4.08, and 4.50 kg m⁻¹ day⁻¹; the heterotrophic denitrification pathway declined faster than the autotrophic pathway. The mechanism of DSR breakdown is as follows. High sulfide concentration inhibits heterotrophic denitrifiers, and the system therefore accumulates nitrite. Autotrophic denitrifiers are then inhibited by the accumulated nitrite, thereby leading to breakdown of the DSR process.     

High-rate denitrifying sulfide removal process in expanded granular sludge bed reactor

An expanded granular sludge bed (EGSB) reactor was adopted to incubate bio-granules that could simultaneously convert 4.8 kg-S m^?3 d^?1 of sulfide in 97% efficiency; 2.6 kg-N m^?3 d^?1 of nitrate in 92% efficiency; and 2.7 kg-C m^?3 d^?1 acetate in 95% efficiency. Mass balance calculation of sulfur, nitrogen, and carbon over the EGSB reactor confirmed the performance results. This noted reactor performance is much higher than those reported in literature. Stoichiometric relation suggests that the nitrate was reduced to nitrite via autotrophic denitrification pathway, then the formed nitrite was converted via heterotrophic denitrification pathway to N₂.     

Potential cause of aerobic granular sludge breakdown at high organic loading rates

Aerobic sludge granules are compact, strong microbial aggregates that have excellent settling ability and capability to efficiently treat high-strength and toxic wastewaters. Aerobic granules disintegrate under high organic loading rates (OLR). This study cultivated aerobic granules using acetate as the sole carbon and energy source in three identical sequencing batch reactors operated under OLR of 9–21.3 kg chemical oxygen demand (COD) m⁻³ day⁻¹. The cultivated granules removed 94–96% of fed COD at OLR up to 9–19.5 kg COD m⁻³ day⁻¹, and disintegrated at OLR of 21.3 kg COD m⁻³ day⁻¹. Most tested isolates did not grow in the medium at >3,000 mg COD l⁻¹; additionally, these strains lost capability for auto-aggregation and protein or polysaccharide productivity. This critical COD regime correlates strongly with the OLR range in which granules started disintegrating. Reduced protein quantity secreted by isolates was associated with the noted poor granule integrity under high OLR. This work identified a potential cause of biological nature for aerobic granules breakdown.     

Characterization of microbial community and kinetics for spent sulfidic caustic applied autotrophic denitrification

Spent sulfidic caustic was applied to sulfur utilizing autotrophic denitrification as the simultaneous source of electron donor and alkalinity. The two experiment set-up of upflow anoxic hybrid growth reactor (UAHGR) and upflow anoxic suspended growth reactor (UASGR) was adopted and nitrate removals were similar in both reactors. Approximately 90% of the initial nitrate was denitrified at nitrate loading rate of 0.15∼0.40 kgNO₃ ⁻/m³·d. The experimental stoichiometric ratio of sulfate production to nitrate removal was ranged from 1.5 to 2.1 mgSO₄ ²⁻/mgNO₃ ⁻. During the operation period, denaturing gradient gel electrophoresis (DGGE) analysis of polymerase chain reaction (PCR)-amplified 16S rDNA fragments for the sludge sample of both reactors showed the change of microbial communities. Thiobacillus denitrificans-like microorganism occupied 28.5% (18 clones) of the 63 clones by cloning the PCR products from the sludge sample of UAHGR. Acidovorax avenae, which can reduce nitrate to nitrogen gas while oxidizing phenol (heterotrophic denitrifier), was also found in 7 clones (11.1%). Although an organic carbon source was not added to the medium, a microorganism (Kaistella koreensis) capable of oxidizing organic compounds was found in 7 clones (11.1%). Therefore, the microbial community of spent sulfidic caustic applied autotrophic denitrification process well corresponds to the substrate components of spent sulfidic caustic. Through the batch cultivation of microorganisms in UAHGR, the microbial kinetic coefficients of spent sulfidic caustic applied autotrophic denitrification were estimated to be μ _max = 0.097 h⁻¹, k _d = 0.0021 h⁻¹, K _s = 200 mgNO₃ ⁻/L, and Y = 0.31 mgMLVSS/mgNO₃ ⁻.     

Biological nitrification–denitrification with alternating oxic and anoxic operations using aerobic granules

Efficient nitrification and denitrification of wastewater containing 1,700 mgl⁻¹ of ammonium-nitrogen was achieved using aerobic granular sludge cultivated at medium-to-high organic loading rates. The cultivated granules were tested in a sequencing batch reactor (SBR) fed with 6.4 or 10.2 kg NH₄⁺-N m⁻³ day⁻¹, a loading significantly higher than that reported in literature. With alternating 2 h oxic and 2 h anoxic operation (OA) modes, removal rate was 45.5 mg NH₄⁺-N g⁻¹ volatile suspended solids⁻¹ h⁻¹ at 6.4 kg NH₄⁺-N m⁻³ day⁻¹ loading and 41.3 ± 2.0 at 10.2 kg NH₄⁺-N m⁻³ day⁻¹ loading. Following the 60 days SBR test, granules were intact. The fluorescence in situ hybridization and confocal laser scanning microscopy results indicate that the SBR-OA granules have a distribution with nitrifers outside and heterotrophs outside that can effectively expose functional strains to surrounding substrates at high concentrations with minimal mass transfer limit. This microbial alignment combined with the smooth granule surface achieved nitrification–denitrification of wastewaters containing high-strength ammonium using aerobic granules. Conversely, the SBR continuous aeration mode yielded a distribution with nitrifers outside and heterotrophs inside with an unsatisfactory denitrification rate and floating granules as gas likely accumulated deep in the granules.     

Bioaugmented sulfur-oxidizing denitrification system with <Emphasis Type="Italic">Alcaligenes defragrans</Emphasis> B21 for high nitrate containing wastewater treatment

The performance of enriched sludge augmented with the B21 strain of Alcaligenes defragrans was compared with that of enriched sludge, as well as with pure Alcaligenes defragrans B21, in the context of a sulfur-oxidizing denitrification (SOD) process. In synthetic wastewater treatment containing 100–1,000 mg NO₃⁻-N/L, the single strain-seeded system exhibited superior performance, featuring higher efficiency and a shorter startup period, provided nitrate loading rate was less than 0.2 kg NO₃⁻-N/m³ per day. At nitrate loading rate of more than 0.5 kg NO₃⁻-N/m³ per day, the bioaugmented sludge system showed higher resistance to shock loading than two other systems. However, no advantage of the bioaugmented system over the enriched sludge system without B21 strain was observed in overall efficiency of denitrification. Both the bioaugmented sludge and enriched sludge systems obtained stable denitrification performance of more than 80% at nitrate loading rate of up to 2 kg NO₃⁻-N/m³ per day.     

Biodegradation and kinetics of aerobic granules under high organic loading rates in sequencing batch reactor

Biodegradation, kinetics, and microbial diversity of aerobic granules were investigated under a high range of organic loading rate 6.0 to 12.0 kg chemical oxygen demand (COD) m⁻³ day⁻¹ in a sequencing batch reactor. The selection and enriching of different bacterial species under different organic loading rates had an important effect on the characteristics and performance of the mature aerobic granules and caused the difference on granular biodegradation and kinetic behaviors. Good granular characteristics and performance were presented at steady state under various organic loading rates. Larger and denser aerobic granules were developed and stabilized at relatively higher organic loading rates with decreased bioactivity in terms of specific oxygen utilization rate and specific growth rate (μ _overall) or solid retention time. The decrease of bioactivity was helpful to maintain granule stability under high organic loading rates and improve reactor operation. The corresponding biokinetic coefficients of endogenous decay rate (k _d), observed yield (Y _obs), and theoretical yield (Y) were measured and calculated in this study. As the increase of organic loading rate, a decreased net sludge production (Y _obs) is associated with an increased solid retention time, while k _d and Y changed insignificantly and can be regarded as constants under different organic loading rates.     

A new method of autotrophic denitrification with spent sulfidic caustic as substrate and alkalinity source

A new method based on sulfide utilizing autotrophic denitrification was adopted to remove nitrate from wastewater and to reuse spent sulfidic caustic containing high sulfide and alkalinity levels. The experiments were performed using a bench-scale upflow anoxic hybrid growth reactor (UAHGR) and an upflow anoxic suspended growth reactor (UASGR) to characterize the stoichiometric relationship between sulfur and nitrate in the process as well as the performance of the reactors. The level of nitrate removal from the UAHGR and UASGR were maintained at over 90% at a nitrate loading rate ranging from 0.15∼0.40 kgNO₃ ⁻/m³·d and no significant nitrite accumulation was observed in either reactor. Although the influent pH values were higher than the optimum range of autotrophic denitrification at 8.7∼10.1, the effluent pH was stable at 7.2∼7.9 due to the production of hydrogen ions during operation. The stoichiometric ratio of sulfate production to nitrate removal was 1.5∼2.1 mgSO₄ ²⁻/mgNO₃ ⁻ in both reactors. A comparison of the reactor performance revealed that the chemical parameters of the UAHGR operation corresponded to a plug flow like type reactor while the chemical parameters of the UASGR operation corresponded to a completely stirred tank reactor like type reactor. UAHGR did not require sludge recycling due to the packed media while UASGR required 300∼700% sludge recycling. Therefore, spent sulfidic caustic could be used in the sulfur utilizing autotrophic denitrification processes as substrate and alkalinity sources.     

Rapid acclimation of methanogenic granular sludge into denitrifying sulfide removal granules

Rapid formation of denitrifying sulfide removal granules is of practical interest to start up an expanded granular sludge bed reactor for wastewater treatment. This study demonstrates that methanogenic granules can be easily acclimated into DSR granules in one day, removing all 1.30 kg m⁻³ d⁻¹ sulfide and converting >90% of 0.56 kg-N m⁻³d⁻¹ nitrate into di-nitrogen gas. Under high loadings, reactor performance, however, declined. Under high loading rates, sulfide first inhibited the heterotrophic denitrifier (Caldithrix sp.), thereby accumulating nitrite in the system; the autotrophic denitrifier (Pseudomonas sp. C23) was then inhibited by accumulated nitrite, leading to breakdown of the entire DSR process.     

Biosorption and biodegradation of pentachlorophenol (PCP) in an upflow anaerobic sludge blanket (UASB) reactor

In order to understand the fate of PCP in upflow anaerobic sludge blanket reactor (UASB) more completely, the sorption and biodegradation of pentachlorophenol (PCP) by anaerobic sludge granules were investigated. The anaerobic granular sludge degrading PCP was formed in UASB reactor, which was seeded with anaerobic sludge acclimated by chlorophenols. At the hydraulic retention time (HRT) of 20–22 h, and PCP loading rate of 200–220 mg l⁻¹ d⁻¹, UASB reactor exhibited good performance in treating wastewater which containing 170–180 mg l⁻¹ PCP and the PCP removal rate of 99.5% was achieved. Sequential appearance of tetra-, tri-, di-, and mono-chlorophenol was observed in the reactor effluent after 20 mg l⁻¹ PCP introduction. Sorption and desorption of PCP on the anaerobic sludge granules were all fitted to the Freundlich isotherm equation. Sorption of PCP was partly irreversible. The Freundlich equation could describe the behavior of PCP amount sorbed by granular sludge in anaerobic reactor reasonably well. The results demonstrated that the main mechanism leading to removal of PCP on anaerobic granular sludge was biodegradation, not sorption or volatization.     

Biotreatment of phenol-contaminated wastewater in a spiral packed-bed bioreactor

A spiral packed-bed bioreactor inoculated with microorganisms obtained from activated sludge was used to conduct a feasibility study for phenol removal. The reactor was operated continuously at various phenol loadings ranging from 53 to 201.4 g m⁻³ h⁻¹, and at different hydraulic retention times (HRT) in the range of 20–180 min to estimate the performance of the device. The results indicated that phenol removal efficiency ranging from 82.9 to 100% can be reached when the reactor is operated at an HRT of 1 h and a phenol loading of less than 111.9 g m⁻³ h⁻¹. At an influent phenol concentration of 201.4 g m⁻³, the removal efficiency increased from 18.6 to 76.9% with an increase in the HRT (20–120 min). For treatment of phenol in the reactor, the maximum biodegradation rate (V _m) was 1.82 mg l⁻¹ min⁻¹; the half-saturation constant (K _s), 34.95 mg l⁻¹.     

Effect of temperature and irradiance on the uptake of ammonium and nitrate by <Emphasis Type="Italic">Laurencia brongniartii</Emphasis> (Rhodophyta,Ceramiales)

The effects of temperature (20, 24 and 28 °C) and irradiance (15 and 40 μmol photon m⁻² s⁻¹) on the nitrate and ammonium uptake rates of the subtropical red alga, Laurencia brongniartii, were investigated to prepare for tank cultivation. Nitrate uptake followed saturation kinetics and was faster at higher irradiances and temperatures. In contrast, ammonium uptake was linear over the experimental range and was not affected by an increase in temperature. A parameter, β, was calculated to compare substrate uptake rates of nitrate along the linear portion of the uptake curve with that of ammonium. For nitrate, β was lower at low irradiance and higher at high irradiance (β = 0.007 ± 0.003 and 0.030 ± 0.002 [μmol N L⁻¹ (μmol N g_ww⁻¹ d⁻)⁻¹], respectively). However, β was 0.023 ± 0.002 and 0.034 ± 0.002 [μmol N L⁻¹ (μmol N g_ww⁻¹ d⁻¹)⁻¹] for ammonium, suggesting a preference for ammonium over nitrate.     

Soybean [Glycine max (L.) merrill] embryogenic cultures: The role of sucrose and total nitrogen content on proliferation

Summary To improve proliferation of soybean cultures in liquid medium, the effects of sucrose; total inorganic nitrogen; content of No₃ ⁻, NH₄ ⁺, Ca²⁺, PO₄ ³⁻, K⁺; NH₄ ⁺/NO₃ ⁻ ratio; and medium osmotic pressure were studied using cv. Jack. Sucrose concentration, osmotic pressure, total nitrogen content, and ammonium to nitrate ratio were found to be the major factors controlling proliferation of soybean embryogenic cultures. Growth decreased linearly as sucrose concentration increased from 29.7 mM to 175.3 mM. A sucrose concentration of 29.2 mM, a nitrogen content of 34.9 mM, at 1 to 4 ammonium to nitrate ratio were found to be optimal for the fastest proliferation of soybean embryogenic cultures. There was no significant effect on proliferation of cultures when concentrations of NH₄ ⁺, Ca²⁺, PO₄ ³⁻, and K⁺ were tested in the range of 3.50 to 10.50, 1.02 to 3.06, 0.68 to 2.04, and 22.30 to 36.70 mM, respectively. The relative proliferation of embryogenic cultures of four soybean genotypes was evaluated in Finer and Nagasawa medium and in the new medium formulation. Despite genotype-specific differences in growth, the genotypes tested showed a biomass increase in the new formulation equal to 278, 269, 170, and 251% for Chapman, F138, Jack, and Williams 82, respectively, relative to their growth on standard FN medium. Due to its lowered sucrose and nitrogen content, we are referring to the new medium as FN Lite.     

Experimental tank cultivation of Porphyra in Israel

Outdoor tank cultivation of several Porphyra (nori) species was carried out from late November 2002 through early May 2003 using 40 L (with a surface of 0.25 m²), 600 L (1 m²), and 24,000 L (30 m²) fiberglass or PVC tanks provided with continuous aeration and seawater flow. Sexual and asexual spores produced from cultured conchocelis and frozen thalli in the laboratory, respectively, were subsequently grown to produce young fronds (ca. 5-10 cm) in an average time of 8 weeks. Growth in outdoor tanks and ponds was possible for a period of up to 20 weeks (i.e. growth season), with yields above 100 g FW m⁻²d⁻¹occurring during 12-14 weeks from late December through late March, when seawater temperatures were below 20 ^∘ C. These yields correlated with the species and depended on the type of tanks in which the algae were cultivated, with the highest yields observed for Porphyra sp. and Porphyra yezoensis when fertilized twice a week with NH₄ Cl and NaH₂ PO₄in 40 L tanks. Calculations of productivity for an entire growth season based on ≥ 100 g FW m⁻²d⁻¹yields exceed the average productivities using seeded nets in open sea, for all Porphyra species tested (0.96-4.06 kg DW m⁻² season⁻¹vs. 0.7-1.0 kg DW m⁻²of net season⁻¹). Therefore, tank cultivation of Porphyra can offer an additional source of nori biomass to international markets. Land-based tank cultivation also offers an environmentally friendly practice that allows for the manipulation of growth conditions to enrich seaweeds with specific, valuable chemicals such as protein and minerals.     

Nitrifying granules cultivation in a sequencing batch reactor at a low organics-to-total nitrogen ratio in wastewater

It is possible to cultivate aerobic granular sludge at a low organic loading rate and organics-to-total nitrogen (COD/N) ratio in wastewater in the reactor with typical geometry (height/diameter = 2.1, superficial air velocity = 6 mm/s). The noted nitrification efficiency was very high (99%). At the highest applied ammonia load (0.3 ± 0.002 mg NH₄⁺–N g total suspended solids (TSS)⁻¹ day⁻¹, COD/N = 1), the dominating oxidized form of nitrogen was nitrite. Despite a constant aeration in the reactor, denitrification occurred in the structure of granules. Applied molecular techniques allowed the changes in the ammonia-oxidizing bacteria (AOB) community in granular sludge to be tracked. The major factor influencing AOB number and species composition was ammonia load. At the ammonia load of 0.3 ± 0.002 mg NH₄⁺–N g TSS⁻¹ day⁻¹, a highly diverse AOB community covering bacteria belonging to both the Nitrosospira and Nitrosomonas genera accounted for ca. 40% of the total bacteria in the biomass.     

An upflow microaerobic sludge blanket reactor operating at high organic loading and low dissolved oxygen levels

The activated sludge process (ASP) has high operational costs due to the need for aeration at dissolved O₂ (DO) levels of ≥2 mg l⁻¹ and high capital costs to construct large reactors due to a low organic loading [typically 1 kg chemical oxygen demand (COD) m⁻³ day⁻¹]. A novel method for improving the energy use and treatment efficiency of the ASP via limited oxygenation (0.4 mg DO l⁻¹) and high organic loading (6.2 kg COD m⁻³ day⁻¹) is proposed based on a laboratory-scale ASP for ammonia-rich industrial wastewaters. The sludge blanket phenomenon and granulation occurred simultaneously in the upflow microaerobic reactor.     

Effect of NaCl on kinetics ofd-glucosamine uptake in yeasts differing in halotolerance

The initial rate ofd-glucosamine uptake by the non-halotolerant yeastSaccharomyces cerevisiae was approximately halved as the apparent half saturation constant (K_m) and the apparent maximum velocity (V_max) changed from 6.6mm to 16.4mm and from 22 μmol · g⁻¹ · min⁻¹ to 16 μmol · g⁻¹ · min⁻¹, respectively, when the salinity in the medium was increased from zerom to 0.68m NaCl. Corresponding changes in a high affinity transport system in the halotolerant yeastDebaryomyces hansenii were from 1.1mm to 4.6mm and from 3.1 μmol · g⁻¹ · min⁻¹ to 4.5 μmol · g⁻¹ · min⁻¹, implying a practically unchanged transport capacity. In 2.7m NaCl, K_m and V_max in this system were 24.5mm and 1.1 μmol · g⁻¹ · min⁻¹, respectively, representing a marked decrease in transport capability. Nevertheless, the degree of affinity in this extreme salinity must still be regarded as noteworthy. In addition to the high affinity transport system inD. hansenii, a low affinity system, presumably without relevance ind-glucosamine transport, was observed.     

Gas phase H<Subscript>2</Subscript>S product recovery in a packed bed bioreactor with immobilized sulfate-reducing bacteria

An N₂ strip gas was used in a packed bed sulfate-reducing bioreactor to recover the dissolved sulfide product and improve sulfate conversion. The highest volumetric productivity obtained was 261 mol H₂S m⁻³ d⁻¹. Lowering the initial pH of the medium from 7 to 6 increased the H₂S content of the strip gas from 3.6 to 5.8 mol%. The ratio of strip gas to liquid flow rates (G/L) was found be to a suitable basis for scaling the process. Calculations indicated that modest G/L values (<10²) were required to recover the residual dissolved sulfide in a downstream stripping column.     

Treatment of food processing wastewater in a full-scale jet biogas internal loop anaerobic fluidized bed reactor

A full-scale jet biogas internal loop anaerobic fluidized bed (JBILAFB) reactor, which requires low energy input and allows enhanced mass transfer, was constructed for the treatment of food processing wastewater. This reactor has an active volume of 798 m³ and can treat 33.3 m³ wastewater per hour. After pre-treating the raw wastewater by settling, oil separating and coagulation-air floating processes, the reactor was operated with a relatively shorter start-up time (55 days). Samples for the influent and effluent of the JBILAFB reactor were taken and analyzed daily for the whole process including both the start-up and stable running periods. When the volumetric COD loading fluctuated in the range of 1.6–5.6 kg COD m⁻³ day⁻¹, the COD removal efficiency, the volatile fatty acid(VFA)/alkalinity ratio, the maximum biogas production and the content of CH₄ in total biogas of the reactor were found to be 80.1 ± 5%, 0.2–0.5, 348.5 m³ day⁻¹ and 94.5 ± 2.5%, respectively. Furthermore, the scanning electron microscope (SEM) results showed that anaerobic granular sludge and microorganism particles with biofilm coexisted in the reactor, and that the bacteria mainly in bacilli and cocci were observed as predominant species. All the data demonstrated that the enhanced mass transfer for gas, liquid and solid phases was achieved, and that the formation of microorganism granules and the removal of inhibitors increased the stability of the system.     

Photoautotrophic micropropagation of Spathiphyllum

In order to maximize yield, Spathiphyllum, an ornamental plant, was cultured in vitro in novel culture vessels termed Vitron. The best growth was obtained by culturing plantlets on sugar-free liquid medium under CO₂ enrichment (3 000 μmol mol⁻¹ 24 h⁻¹ d⁻¹) at a low photon flux density (PPFD of 45 μmol m⁻² s⁻¹), suggesting that the novel Vitron culture system is suitable for the photoautotrophic micropropagation of Spathiphyllum.