Growth of Salvinia molesta as affected by water temperature and nutrition I. Effects of nitrogen level and nitrogen compounds

The growth of Salvinia molesta D.S. Mitchell was studied in a greenhouse using controlled-temperature water-baths at 16, 19 and 22°C and 4 different nitrogen compounds (NO₃^?, NH₄⁺, NH₄NO₃ and urea) at levels up to 60 mg N l^?1. Little growth occurred at 16°C even if 20 mg N l^?1 was supplied together with other nutrients including phosphorus (2 mg H₂PO₄-P l^?1). The highest relative growth rate and total dry matter production occurred at 22°C when plants were supplied with 20 mg NH₄-N l^?1. At this temperature, the NH₄⁺ ion was superior to the NO₃^? ion or urea as a nitrogen source (almost doubling the biomass), but was not significantly better than NH₄NO₃. Over a period of 19 days for plants receiving 0.02 mg NH₄-N l^?, biomass increased 4-fold at 16°C, 9-fold at 19°C and 10-fold at 22°C. In contrast, for plants receiving 20 mg NH₄-N l^?1, biomass increased 4-fold at 16°C, 18-fold at 19°C and 38-fold at 22°C.     

九龙江河口区养虾塘沉积物-水界面营养盐交换通量特征

通过对九龙江河口区陆基养虾塘水样和沉积物样品采集分析及结合室内模拟实验,探讨了虾塘在不同养殖阶段沉积物-水界面营养盐通量时间变化特征及其主要影响因素。虾塘沉积物向上覆水体释放NO_x~--N(NO_2~--N和NO_3~--N)、NH_4~+-N和PO_4~(3-)-P能力均呈现随养殖时间推移而降低的特征。沉积物在养殖中期和后期分别呈现对上覆水体NO_x~--N和PO_4~(3-)-P的吸收现象,但总体表现为释放(平均通量分别为(1.87±1.15)、(1.58±0.52)mg m~(-2)h~(-1)和(1.22±0.62)mg m~(-2)h~(-1))。沉积物-水界面溶解无机氮交换以NH_4~+-N为主(沉积物平均释放通量为(46.18±13.82)mg m~(-2)h~(-1))。沉积物间隙水与上覆水间的营养盐浓度差(梯度)及温度对上述交换通量的时间动态特征具有重要调控作用。研究结果表明养殖初期或中期沉积物较高的无机氮(尤其是NO_2~--N和NH_4~+-N)释放是养殖塘水质恶化的一个极具潜力的污染内源,可能会对虾的健康生长产生负面效应,控制沉积物无机氮释放是养虾塘养殖初期和中期重要的日常管理活动之一。     

FACTORS REGULATING THE SPATIAL DISTRIBUTION OF THE FILAMENTOUS ALGA PITHOPHORA OEDOGONIA (CHLOROPHYCEAE) IN AN INDIANA LAKE1

Pithophora oedogonia (Mont.) Wittr. biomass in Surrey Lake, Indiana was greater in the littoral than in the pelagial region. Although mean soluble reactive phosphorus concentrations did not differ between the two areas, nitrate concentrations were almost six times higher in the cove than in the open water. Using laboratory cultures of Pithophora, the half saturation constant (K_s at 20° C relating filament growth to external concentrations of nitrate-nitrogen was determined to be 1.23 mg L^?1 (=88 μM)and for phosphate-phosphorus, 0.1 mg L^?1 (=3.22 μM). These values were used to calculate a NO₃-N/PO₄-P atomic ratio of 27.6. Comparison of this value with NO₃-N/PO₄-P ratios in Surrey Lake showed that nitrogen limiting conditions were prevalent in the open water section of the lake. Alkaline phosphatase and dark ammonia uptake analyses on field collected filaments from the shallow and deep water sections confirmed the hypothesis that nitrate is the major factor limiting growth of Pithophora in Surrey Lake.     

Growth of Salvinia molesta as affected by water temperature and nutrition II. Effects of phosphorus level

The growth of Salvinia molesta D.S. Mitchell, as affected by phosphorus supply and water temperature, was studied in a greenhouse using controlled water temperature baths at 16, 19 and 22°C. For significant responses to the phosphorus treatments to be obtained it was found necessary to use P-deficient plant propagules (containing <0.01% P on a dry matter basis). For these plants the highest relative growth rate and dry matter production occurred at 22°C when they received 10.01 mg PO₄P l^?1, but this was not significantly different from that of plants receiving 1.01 mg PO₄-P l^?1. Over a period of 21 days for plants receiving 0.02 mg PO₄ l^?1 the biomass increased 4-fold at 19°C and 6-fold at 22°C. In contrast, for plants receiving 10.01 mg PO₄-P l^?1 biomass was increased 20-fold at 19°C and 32-fold at 22°C. At the latter temperature, when receiving 60.01 mg PO₄-P l^?1, plants concentrated up to 1.3% of phosphorus on a dry matter basis, suggesting a possible use as a biological filter and purifier of contaminated waters.     

Two different methods of evaluating nutrient limitations of periphyton bioassays,using water from the River Rhine and eight of its tributaries

An investigation was untertaken to evaluate the nutrient status of the River Rhine (two stations) and eight of its tributaries (total of ten samplings). Determinations of the following inorganic substances were made: PO₄ ^?3-P; NO₃ ^?-N; NO₂ ^?-N; NH₄ ⁺ -N and Cl^?. In addition, pH and carbonate alkalinity were measured. Bioassays to obtain the algal growth potential (AGP) were carried out using periphyton from the River Rhine. A linear relationship could be established between NO₃ ^?-N and the AGP, while the AGP showed a non-linear dependence on the PO₄ ^3?-P concentration. The critical N/P ratio for N or P limitation of the algal growth in bioassays was evaluated graphically and by calculation. The results of the two methods are in good agreement: N is the limiting factor at NO₃ ^?N/PO₄ ^3?-P ratios less than 10, while P is limiting at ratios greater than 20. At values between 10 and 20 neither N nor P can be supposed with certainty to be limiting.     

Algal biomass production,N uptake and N2 fixation in a synthetic medium

An experiment was conducted in the growth chamber to quantify the biomass production, N removal and N₂ fixation from a synthetic medium by Chlamydomonas reinhardtii and Anabaena flos-aquae. Nitrogen was supplied at a concentration of 100 mg liter⁻¹ of NO₃⁻−¹⁵N and NH₄⁺−¹⁵ (3·5 atom %), respectively. After 21 days Chlamydomonas reinhardtii removed an average of 83·8 and 78·7 mg N liter⁻¹ as NO₃⁻ and NH₄⁺, respectively. Averages of 0·89 and 0·71 g liter⁻¹ (first batch), 1·63 and 0·95 g liter⁻ (second batch) algal biomass were collected from NO₃⁻ and NH₄⁺ media, respectively. Uptake rates of 0·11 mg ¹⁵N g⁻¹ algae day⁻¹ from NO₃⁻ medium and 0·10 mg ¹⁵N g⁻¹ algae day⁻¹ from NH₄⁺ medium were calculated. Algal cells grown in NO₃⁻ and NH₄⁺ medium contained 71 and 65 g N kg⁻¹ (first batch), 39 and 58 g N kg⁻¹ (second batch), respectively. Anabaena flos-aquae produced averages of 0·58 and 0·46 g liter⁻¹ (first batch), 0·55 and 0·48 g liter⁻¹ (second batch) after 14 days of growth from NO₃⁻ and NH₄⁺ media, respectively. Blue-green algal biomass contained higher N (81–98 g kg⁻¹) than green algae. Isotope dilution method for determining N₂ fixation indicated that 55% and 77% of total N of blue-green algae grown in NO₃⁻ and NH₄⁺ media, respectively, was derived from the atmosphere.     

胶州湾夏秋季大气湿沉降中的营养盐及其入海的生态效应

为揭示大气湿沉降对胶州湾营养盐的输送通量及其生态效应,分别于2015年6—8月(夏季)、9—11月(秋季)采集胶州湾降水样品,测定了降水中不同形态N、P、Si的浓度。结果表明,降水中不同形态营养盐的浓度变化较大,且均与降水量呈负相关关系,其中NH4-N和NO3-N的浓度较高,溶解有机氮(DON)占溶解态总氮(DTN)含量的25.9%,而NO_2-N,PO_4-P和SiO_3-Si的浓度均很低。溶解无机氮(DIN)、DON、PO_4-P以及SiO_3-Si的湿沉降通量分别为141.7、61.87、0.35 mmol m~(-2)a~(-1)和0.12 mmol m~(-2)a~(-1)。受降水量和营养物质来源制约,各项营养盐湿沉降通量时间变化显著。农业活动导致的无机氮排放构成了胶州湾湿沉降DIN的主要来源。大气湿沉降DIN、DON、PO_4-P和SiO_3-Si分别占胶州湾总输入负荷的9.04%、10.24%、0.57%和0.17%,湿沉降输入的PO_4-P在夏、秋季分别可以支持0.575 mgC m~2d~(-1)和1.42 mg C m~2d~(-1)的新生产力;雨水中DIN/P比值高达1 617,突发性强降雨带来的营养盐输入会加剧表层水体的P限制和Si限制,对胶州湾浮游植物群落结构和粒级结构产生重要影响。大气湿沉降是胶州湾生源要素生物地球化学过程的重要一环,对营养物质收支的贡献及可能引发的生态效应不容忽视。     

Chemical composition and nutrient loading by precipitation in the Trachypogon savannas of the Orinoco llanos, Venezuela

Samples of bulk precipitation were collected in the Trachypogon savanna, Calabozo, Venezuela, during three consecutive years. In the first year, rain samples were taken daily; in the following years the samples were grouped on a monthly basis. In addition, samples of dry deposition were collected during the dry seasons. All samples were analyzed for the following water soluble cations and anions: P0₄-P, S0₄-S, N0₃-N, NH₄-N, Ca⁺², Mg⁺², K⁺, Na⁺ and H⁺. The mean annual input rate of chemical constituents (Kg ha^-1 year^-1) was: PO₄-P (0.42); SO₄-S (2.62); NO₃-N (0.21); NH₄-N (2.03); Ca₊₂ (3.50); Mg⁺² (11.31); K⁺ (3.60); Na⁺ (5.93) and H⁺ (0.03). The total mean input of particulate material to the savanna during the dry season was 2.06 Kg ha^-1 year^-1, with a soluble fraction of 30%. Possible sources of nutrients input were analyzed.     

Evaluation of leaf analysis as a guide to nitrogen and phosphorus fertilization of potato (Solanum tuberosum L.)

Summary Recently matured leaf samples were collected, at 45, 60 and 75 days after planting, from potato (Solanum tuberosum L.) plants of cultivar Kufri sindhuri grown with varying levels of nitrogen (0, 60, 120, 180 and 240 kg N/ha) and phosphorus (0, 60, 120 and 180 kg P₂O₅/ha) on loam soil at Pantnagar. They were separated into petiole, midrib and leaf-lets and analysed for NO₃-N content. Petiole samples were also analysed for PO₄-P content. Nitrogen application increased the NO₃-N content of all the leaf components. P application increased the PO₄-P content in petiole. NO₃-N content and PO₄-P content in leaf tissues were positively correlated with final tuber yield. The association of NO₃-N content of petiole with the final tuber yield was very consistant. Hence this proved to be the best indicator tissue for reflecting the nitrogen status of the plant, particularly at 45 days after planting. NO₃-N content of midrib, at this stage, was also a good indication of nitrogen nutrition status of the plant. PO₄-P content of petiole at 45 days after planting was a good indication of nutritional status of plant with respect to phosphorus. The critical concentration of NO₃-N in petioles of 45 days old plants was in the range of 9100–9600 ppm. The corresponding range for midrib was 3000 to 3900 ppm. The critical concentration of PO₄-P for petioles of 45 days old plants was 2250 ppm.Publication No 796 under journal series of the G.B. Pant University of Agriculture and Technology, Experiment Station, Pantnagar.Junior Agronomist, Indian Institute of Horticultural Research, Bangalore-6, India.Junior Agronomist, Indian Institute of Horticultural Research, Bangalore-6, India.     

Effect of a modular extensive green roof on stormwater runoff and water quality

Runoff quantity and quality from a 248 m² extensive green roof and a control were compared in Connecticut using a paired watershed study. Weekly and individual rain storm samples of runoff and precipitation were analyzed for TKN, NO₃ + NO₂-N, NH₃-N, TP, PO₄-P, and total and dissolved Cu, Pb, Zn, Cd, Cr, and Hg. The green roof watershed retained 51.4% of precipitation during the study period based on area extrapolation. Overall, the green roof retained 34% more precipitation than predicted by the paired watershed calibration equation. TP and PO₄-P mean concentrations in green roof runoff were higher than in precipitation but lower than in runoff from the control. The green roof was a sink for NH₃-N, Zn, and Pb, but not for TP, PO₄-P, and total Cu. It also reduced the mass export of TN, TKN, NO₃ + NO₂-N, Hg, and dissolved Cu primarily through a reduction in stormwater runoff. Greater than 90% of the total Cu, Hg, and Zn concentrations in the green roof runoff were in the dissolved form. The growing media and slow release fertilizer were probable sources of P and Cu in green roof runoff. Overall, the green roof was effective in reducing stormwater runoff and overall pollutant loading for most water quality contaminants.     

EFFECT OF SODIUM AND NITRATE ON GROWTH OF ANABAENA FLOS-AQUAE (CYANOPHYTA)1

The growth response of a nitrogen fixing cyanobacterium, Anabaena flos-aquae (Lyng) Bréb., to sodium and nitrate was examined in batch culture under controlled laboratory conditions. Sodium (range 0-12 mg Na⁺· L^-1) enhanced growth of the cyanobacterium under nitrate-sufficient (5.7 mg NO³-N · L^-1) but not nitrate-limited (0.49 mg NO³- N · L^-1) conditions. The magnitude of the growth response was related to the nutritional history of the culture. No significant effect of sodium on nitrate utilization was observed. The increase in ambient sodium levels in many lakes may provide a competitive advantage to cyanobacteria.     

Evaluation of microalgal consortia for treatment of primary treated sewage effluent and biomass production

The present investigation was aimed towards analyzing the potential of consortia of native filamentous microalgal strains (MC2), native unicellular microalgal strains (MC3), and selected microalgae from germplasm (MC1) in terms of nutrient removal, water quality improvement, and biomass production using primary treated sewage water. Highest NO₃-N (90 %) and PO₄-P (97.8 %) removal was obtained with MC2-inoculated sewage water. Highest decrease in total dissolved solids to 806 from 1,120 mg L^?1 and highest increase in dissolved oxygen of 9.0 from 0.4 mg L^?1 were obtained using MC2-inoculated sewage water on the sixth day. The biomass production was also highest in MC2 (1.07 g L^?1) followed by MC1 and MC3 (0.90 and 0.94 g L^?1, respectively) on the sixth day. The consortium of filamentous strains from native environment not only proved promising in nutrient removal efficiency but also led to enhanced biomass. The present study highlighted the utility of such a consortium for sewage wastewater treatment and the promise of sewage water as a growth medium for biomass production.     

Effects of macro elements and nitrogen source on adventitious root growth and ginsenoside production in ginseng (Panax ginseng C. A. Meyer)

We investigated the effects on ginseng adventitious root growth and ginsenoside production when macro-element concentrations and nitrogen source were manipulated in the culture media. Biomass growth was greatest in the medium supplemented with 0.5-strength NH₄PO₃, whereas ginsenoside accumulation was highest (9.90 mg g^-1 DW) in the absence of NH₄PO₃. At levels of 1.0-strength KNO₃, root growth was maximum, but a 2.0 strength of KNO₃ led to the greatest ginsenoside content (9.85 mg g^-l). High concentrations of MgSO₄ were most favorable for both root growth and ginsenoside accumulation (up to 8.89 mg g^-1 DW). Root growth and ginsenoside content also increased in proportion to the concentration of CaCI₂ in the medium, with the greatest accumulation of ginsenoside (8.91 mg g^-1 DW) occurring at a 2.0 strength. The NH₄/NO₃ ^-- ratio also influenced adventitious root growth and ginsenoside production; both parameters were greater when the NO₃ ^- concentration was higher than that of NH₄ ⁺. Maximum root growth was achieved at an NH₄ ⁺/NO₃ ^- ratio of 7.19/18.50, while ginsenoside production was greatest (83.37 mg L^-1) when NO₃ ^- was used as the sole N source.     

马尾松组培苗对氮素形态的生长响应

马尾松属高氮需求树种,然而在苗木培育中马尾松对氮素,尤其是不同形态氮素的需求尚不明确.该文以马尾松组培苗为试验材料,采用基质培养方法,针对硝态氮、铵态氮两种氮素形态均分别设置了2、4、8、16 mmol·L-14个处理,以不添加氮素为对照,对苗木的高径生长、根构型参数(总根长、总表面积、总体积、平均直径和根尖数)以及生...     

Water quality of the Modder River,South Africa

The Modder River is a relatively small river in the central region of the Free State Province, South Africa and has a mean annual runoff of 184 × 106m³. Botshabelo is a city, which has developed in the catchment area of the river, and its sewage outflows are discharged into the Klein Modder, a tributary of the Modder River. This study was conducted in order to determine the influence of Botshabelo's sewage outflow on the water quality of the river. It was determined that the Modder and Klein Modder Rivers do not generally follow distinctive seasonal patterns in terms of chemical parameters, although NO₃-N and PO₄-P concentrations usually increase with increasing flow and conductivity decreases with increasing flow. Physical parameters such as turbidity, flow and temperature did however follow distinctive seasonal patterns from February 1996 to December 1997, as did phytoplankton growth. Low chlorophyll-a concentrations were exhibited in the winter and higher concentrations during spring. In the Klein Modder River, algal blooms occurred more frequently, and the algal biomass was higher than in the Modder River. This could be ascribed to the higher nutrient concentrations and lower flow velocities in the former. The inflow of the Klein Modder River into the Modder River caused on average, 112% increase in PO₄-P, 171% increase in NO₃-N, 50% increase in chlorophyll-a concentration, and 230% increase in E. coli counts.     

海水中藻菌共培养体系对碳氮磷的吸收转化

海洋环境中,细菌和微藻之间的物质交换是生源要素在自然界中迁移转化的重要方式。为进一步了解生源要素的生物地球化学循环,在实验室模拟条件下,研究了共培养体系中营养盐和有机物在细菌和微藻之间的转换。通过纯培养中肋骨条藻(Skeletonema costatum)、东海原甲藻(Prorocentrum donghaiense)、天然海水中的细菌以及藻菌混合培养,分析了营养盐和有机物随藻菌生物量的变化情况,并计算了溶解有机碳(DOC)和溶解有机氮(DON)的浓度比值[(DOC/DON)a]。结果发现,在共培养体系中,细菌对中肋骨条藻的生长有抑制作用,对东海原甲藻影响不明显;中肋骨条藻有利于细菌生长,东海原甲藻抑制细菌生长,这种不同可能与微藻的粒径有关。海洋细菌在2种藻的指数生长均期均会促进微藻吸收氨氮(NH_4-N),但在生长末期NH_4-N以释放为主。硝氮(NO_3-N)的浓度与藻的生长呈负相关,但在衰亡期NO_3-N略有增加,表明NO_3-N再生所需时间较长。细菌对硝氮的吸收量较少,但对其再生有贡献。细菌和中肋骨条藻对磷酸盐(PO_4-P)的吸收存在竞争,但与东海原甲藻的竞争关系不明显。不同培养体系中DOC浓度变化不同,在藻菌共培养体系中增加较快,纯藻培养体系中增加缓慢,在纯菌培养体系中缓慢减少。通过对DOC与DON浓度比值的分析,发现用判断颗粒有机碳(POC)来源的方法可以分析DOC的来源。     

Efficient itaconic acid production by Aspergillus terreus: Overcoming the strong inhibitory effect of manganese

Itaconic acid (IA), a building block platform chemical, is produced industrially by Aspergillus terreus utilizing glucose. Lignocellulosic biomass can serve as a low cost source of sugars for IA production. However, the fungus could not produce IA from dilute acid pretreated and enzymatically saccharified wheat straw hydrolyzate even at 100-fold dilution. Furfural, hydroxymethyl furfural and acetic acid were inhibitory, as is typical, but Mn²⁺ was particularly problematic for IA production. It was present in the hydrolyzate at a level that was 230 times over the inhibitory limit (50 ppb). Recently, it was found that PO₄³⁻ limitation decreased the inhibitory effect of Mn²⁺ on IA production. In the present study, a novel medium was developed for production of IA by varying PO₄³⁻, Fe³⁺ and Cu²⁺ concentrations using response surface methodology, which alleviated the strong inhibitory effect of Mn²⁺. The new medium contained 0.08 g KH₂PO₄, 3 g NH₄NO₃, 1 g MgSO₄·7H₂O, 5 g CaCl₂·2 H₂O, 0.83 mg FeCl₃·6H₂O, 8 mg ZnSO₄·7H₂O, and 45 mg CuSO₄·5H₂O per liter. The fungus was able to produce IA very well in the presence of Mn²⁺ up to 100 ppm in the medium. This medium will be extremely useful for IA production in the presence of Mn²⁺. This is the first report on the development of Mn²⁺ tolerant medium for IA production by A. terreus.     

The study of phosphorus and nitrogen fluxes in enriched isolation columns

Isolation columns positioned in approximately 3.5 m of water in Midmar Dam, were enriched with PO₄-P and NO₃-N, singly and in combination. Increased chlorophyll concentration resulted from all enrichment treatments. Responses showed marked seasonality and varied in intensity and duration. Despite the PO₄-P fixing potential of the sediments, SRP concentrations increased markedly at times and possible reasons for this discussed. The marked response to addition of NO₃-N alone suggested a flux of phosphorus from the sediment through the water to the algal component. The significance of periphyton within the columns is discussed.     

Effect of influent substrate ratio on anammox granular sludge: performance and kinetics

Effect of influent substrate ratio on anammox process was studied in sequencing batch reactor. Operating temperature was fixed at 35 ± 1 °C. Influent pH and hydraulic retention time were 7.5 and 6 h, respectively. When influent NO₂ ^?-N/NH₄ ⁺-N was no more than 2.0, total nitrogen removal rate (TNRR) increased whereas NH₄ ⁺-N removal rate stabilized at 0.32 kg/(m³ d). ΔNO₂ ^?-N/ΔNH₄ ⁺-N increased with enhancing NO₂ ^?-N/NH₄ ⁺-N. When NO₂ ^?-N/NH₄ ⁺-N was 4.5, ΔNO₂ ^?-N/ΔNH₄ ⁺-N was 1.98, which was much higher than theoretical value (1.32). The IC₅₀ of NO₂ ^?-N was 289 mg/L and anammox activity was inhibited at high NO₂ ^?-N/NH₄ ⁺-N ratio. With regard to influent NH₄ ⁺-N/NO₂ ^?-N, the maximum NH₄ ⁺-N removal rate was 0.36 kg/(m³ d), which occurred at the ratio of 4.0. Anammox activity was inhibited when influent NH₄ ⁺-N/NO₂ ^?-N was higher than 5.0. With influent NO₃ ^?-N/NH₄ ⁺-N of 2.5–6.5, NH₄ ⁺-N removal rate and NRR were stabilized at 0.33 and 0.40 kg/(m³ d), respectively. When the ratio was higher than 6.5, nitrogen removal would be worsened. The inhibitory threshold concentration of NO₂ ^?-N was lower than NH₄ ⁺-N and NO₃ ^?-N. Anammox bacteria were more sensitive to NO₂ ^?-N than NH₄ ⁺-N and NO₃ ^?-N. TNRR would be enhanced with increasing nitrogen loading rate, but sludge floatation occurred at high nitrogen loading shock. The Han-Levenspiel could be applied to simulate nitrogen removal resulting from NO₂ ^?-N inhibition.     

Influence of Agapanthus africanus on nitrification in a vertical subsurface flow constructed wetland

The aim of this study is to evaluate the influence of Agapanthus africanus (A. africanus) on nitrification in a vertical subsurface flow constructed wetlands (VSSFs) system. Two lab-scale VSSFs were operated: a) one was planted with A. africanus (vertical flow planted, VFP), and b) the other was unplanted (vertical flow control, VFC). The operation strategy was divided into three phases and consisted of increasing the ammoniacal nitrogen loading rate (ALR) (Phase I: 1.4; Phase II: 2.4; Phase III: 4.4 g NH₄⁺-N·m^?2·d^?1). Nitrification was evaluated in the system at two different depths in the VSSFs (30.5 cm and 60.3 cm, from the top of the system).

The removal efficiencies of COD, BOD₅, TP, and PO₄^?3-P were above 40% in the VFP and VFC during all operation. The mean removal efficiencies of NH₄⁺-N were above 70%. Nitrification was the principal NH₄⁺-N removal mechanism in both systems and transformed more than 50% of the NH₄⁺-N to NO₃^?-N. In terms of the effect of A. africanus on NH₄⁺-N removal during the three operational phases, nonsignificant differences between the two VSSFs were noted (p > 0.05). Thus, A. africanus did not influence nitrification. Finally, the analysis at different depths showed that nitrification occurred in the upper 30.5 cm.