Jin et al 2026b - Revision history

Tomamil: Tomamil moved page Review 699279737891 to Jin et al 2026b

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Tomamil moved page Review 699279737891 to Jin et al 2026b

TianJin at 12:02, 4 March 2026

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TianJin at 03:44, 4 March 2026

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TianJin at 05:27, 3 March 2026

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Ashley at 07:14, 26 February 2026

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NZHNG at 01:34, 26 February 2026

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NZHNG at 01:31, 26 February 2026

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NZHNG at 01:27, 26 February 2026

2026-02-26T01:27:48Z

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 ==4.0 DATA AND RESULTS==
 [[File:MW update.png|none|thumb|561x561px|'''Figure 1. Trial 1 Mean Fresh Weight with Time.''' (a) Trial 1: as a progression from Day 0 to Day 28 and (b) Day 0, Day 14, and Day 28 on individual dates. Error bar values represent mean ±STDEV. Statistical significance was not calculated due to insufficient sample sizes in some treatment groups. (c) Trial 2: as a progression from Day 0 to Day 24 and (d) Day 0, Day 14, and Day 24 on individual dates. Error bar values represent mean ±SEM. Statistically significant comparisons (p<0.05) are represented with different letters.]]
-In Trial 1, Fig. 1a & Fig. 1b, total fresh weight generally decreased over time across most treatment groups, with the exception of the 0.002M NO₃⁻ treatment (0.3499 g by Day 28). Ammonium-containing treatments showed a substantial decline through Day 28. In Trial 2, Fig. 1c & Fig. 1d, total fresh weight changed significantly over time depending on the source of nitrogen. Plants treated with sole nitrate had the greatest increase in biomass from Day 0 (0.546 g) to Day 24 (1.730), with a significant increase by Day 14, resulting in the highest overall fresh weight. Treatments containing ammonium showed reduced growth rates over time. Both the 1:3 and 1:1 NH₄⁺ to NO₃⁻ treatments initially increased by Day 14 (0.999 g and 0.649 g, respectively) and declined by Day 24. By Days 14 and 24, nitrate-induced plants were significantly greater in biomass than ammonium-induced plants, suggesting that nitrate supports biomass accumulation while ammonium negatively affects long-term growth.[[File:DW update.png|none|thumb|571x571px|'''Figure 2. Dry Weight Per Treatment Group.''' (a) Trial 1: the end samples per treatment varied: 0.002 M NO₃⁻ (6 samples), 1:3 (1 sample), 1:1 (1 sample), 3:1 (0 samples), and 0.002 M NH₄⁺ (3 samples). (b) Trial 2: the end samples per treatment varied among water (10 samples), 0.002 M NO3- (18 samples), 1:3 (8 samples), 1:1 (3 samples), 3:1 (3 samples), and 0.002 M NH₄⁺ (0 samples).]]Final dry weight measurements support the trend observed in Fig. 1, indicating that nitrogen form influenced biomass accumulation. In Fig. 2a, The 0.002M NO₃⁻ had the highest overall dry weight (0.0429 g), followed by the 0.002M NH₄⁺ treatment (0.0309 g). Treatments containing both nitrate and ammonium yielded considerably lower dry mass. The 3:1 NH₄⁺ to NO₃⁻ treatment group had no remaining samples at Day 28, preventing measurement of dry weight. In Trial 2, Plants treated with 0.002M NO₃⁻ produced the greatest final dry weight (0.1231 g) in comparison to all other treatment groups. The control showed less dry weight than the sole nitrate treatment (0.0953 g). Treatments containing ammonium yielded comparably lower dry weights (1:3 NH₄⁺ to NO₃⁻, 0.0292 g; 1:1 NH₄⁺ to NO₃⁻, 0.0055 g; 3:1 NH₄⁺ to NO₃⁻, 0.0052 g). The 0.002M NH₄⁺ treatment produced no measurable dry weight by the end of the trial.
+In Trial 1, Fig. 1a & Fig. 1b, total fresh weight generally decreased over time across most treatment groups, with the exception of the 0.002M NO₃⁻ treatment (0.3499 g by Day 28). Ammonium-containing treatments showed a substantial decline through Day 28. In Trial 2, Fig. 1c & Fig. 1d, total fresh weight changed significantly over time depending on the source of nitrogen. Plants treated with sole nitrate had the greatest increase in biomass from Day 0 (0.546 g) to Day 24 (1.730), with a significant increase by Day 14, resulting in the highest overall fresh weight. Treatments containing ammonium showed reduced growth rates over time. Both the 1:3 and 1:1 NH₄⁺ to NO₃⁻ treatments initially increased by Day 14 (0.999 g and 0.649 g, respectively) and declined by Day 24. By Days 14 and 24, nitrate-induced plants were significantly greater in biomass than ammonium-induced plants, suggesting that nitrate supports biomass accumulation while ammonium negatively affects long-term growth.
-[[File:MRL Updated.png|none|thumb|576x576px|'''Figure 3. Mean Root Length with Time'''. (a) Trial 1: as a progression from Day 0 to Day 28 and (b) Day 0, Day 14, and Day 28 on individual dates. Error bar values represent mean ±STDEV. Statistical significance was not calculated due to insufficient sample sizes in some treatment groups. (c) Trial 2: as a progression from Day 0 to Day 24 and (d) Day 0, Day 14, and Day 24 on individual dates. Error bar values represent mean ±SEM. Statistically significant comparisons (p<0.05) are represented with different letters.]]In Trial 1, Fig. 3a & Fig. 3b, mean root length generally decreased over time across all nitrogen treatments. The 0.002M NO₃⁻ group had the highest average root length by Day 28 compared to other treatments. In Trial 2, Fig. 3c & Fig. 3d, plants grown with 0.002M NH₄⁺ initially exhibited the greatest root elongation, having the highest mean root length by Day 14 (1.100 cm), but declined by Day 24. In contrast, 0.002M NO₃⁻ showed steady root growth, resulting in one of the significantly higher mean root lengths by Day 24 (1.436 cm). The control and 1:3 NH₄⁺ to NO₃⁻ also indicated relatively high root elongation by Day 24 (0.750 and 1.060 cm, respectively). Treatments containing higher amounts of ammonium (1:1 and 3:1 NH₄⁺ to NO₃⁻) consistently produced shorter roots (0.267 and 0.367 cm, respectively).
+[[File:DW update.png|none|thumb|571x571px|'''Figure 2. Dry Weight Per Treatment Group.''' (a) Trial 1: the end samples per treatment varied: 0.002 M NO₃⁻ (6 samples), 1:3 (1 sample), 1:1 (1 sample), 3:1 (0 samples), and 0.002 M NH₄⁺ (3 samples). (b) Trial 2: the end samples per treatment varied among water (10 samples), 0.002 M NO3- (18 samples), 1:3 (8 samples), 1:1 (3 samples), 3:1 (3 samples), and 0.002 M NH₄⁺ (0 samples).]]
 [[File:TLA update 2.png|none|thumb|599x599px|'''Figure 4. Total Leaf Amount'''. (a) Trial 1: as a progression from Day 0 to Day 28 and (b) Day 0, Day 14, and Day 28 on individual dates. Error bar values represent mean ±STDEV. Statistical significance was not calculated due to insufficient sample sizes in some treatment groups. (c) Trial 2: as a progression from Day 0 to Day 24 and (d) Day 0, Day 14, and Day 24 on individual dates. Error bar values represent mean ±SEM. Statistically significant comparisons (p<0.05) are represented with different letters.]]
 In Trial 1, Fig. 4a & Fig. 4b, total leaf amount generally declined with combined nitrate and ammonium treatments. By Day 28, 0.002M NO₃⁻ showed the greatest increase in total leaf count relative to other treatments (45) followed by 0.002M NH₄⁺ (17). In contrast, combined treatments (1:3, 1:1, and 3:1 NH₄⁺ to NO₃⁻) showed reductions in leaf number over time (1, 4, and 0 by Day 28).In Trial 2, Fig. 4c & Fig. 4d, total leaf amount increased over time, primarily in the 0.002M NO₃⁻ group, showing a statistically significant increase from Day 0 (42) to Day 24 (140), and had the highest overall leaf number. The control group showed a moderate increase by Day 14, while the 1:3 NH₄⁺ to NO₃⁻ group showed some growth at Day 14 (77), then decreased by nearly 50% at Day 24 (38). Conversely, the 1:1 and 3:1 NH₄⁺ to NO₃⁻ treatments significantly reduced total leaf amount over time, and the 0.002M NH₄⁺ consistently had the lowest leaf count.
 [[File:Survival percentage update.png|none|thumb|600x600px|'''Figure 6. Survival Percentage with Time.''' (a) Trial  1: the sample size is a percentage from Day 0 to Day 28. (b) Trial 2: the sample size is a percentage from Day 0 to Day 24]]
-Similar to Figures 1-5, survival rates depended on the nitrogen sources over time. In Trial 1, Fig. 6a, survival generally remained stable in the 0.0022M NO₃⁻ treatment, maintaining 100% survival by Day 28. The 0.002M NH₄⁺ treatment group initially increased to 116.67% and remained stable through Day 11, but declined to 50% of the original sample size at Day 14. Conversely, mixed nitrogen sources showed a reduction in survival percentage over time, with 3:1 NH₄⁺ to NO₃⁻ treatment reaching complete mortality by Day 28. In Trial 2, Fig. 6b, plants treated with sole NO₃⁻ maintained the highest survival throughout the trial, at 150% by Day 24. The control and 1:3 NH₄⁺ to NO₃⁻ fluctuated in the survival number but generally declined towards the end. Conversely, higher ammonium treatments, 1:1 and 3:1 NH₄⁺ to NO₃⁻, showed steep declines following Day 16, and sole NH₄⁺ treatment exhibited complete mortality. Overall, nitrate treatments sustained plant viability, while increasing ammonium treatments negatively influenced survival over time.[[File:Images update.png|none|thumb|589x589px|'''Figure 7. Trial 1: Image of Plants Taken on (a) Day 0, (b) Day 7, (c) Day 14, (d) Day 21, and (e) Day 28.''' Images of plants were generally taken every Monday throughout the study duration.]]Algae growth was seen in 0.002M NO₃⁻, 1:1 NH₄⁺ to NO₃⁻, and 3:1 NH₄⁺ to NO₃⁻. Comparing treatment group photos from Day 0 (A) to Day 28 (E), sample sizes decreased as plant conditions worsened.
+Similar to Figures 1-5, survival rates depended on the nitrogen sources over time. In Trial 1, Fig. 6a, survival generally remained stable in the 0.0022M NO₃⁻ treatment, maintaining 100% survival by Day 28. The 0.002M NH₄⁺ treatment group initially increased to 116.67% and remained stable through Day 11, but declined to 50% of the original sample size at Day 14. Conversely, mixed nitrogen sources showed a reduction in survival percentage over time, with 3:1 NH₄⁺ to NO₃⁻ treatment reaching complete mortality by Day 28. In Trial 2, Fig. 6b, plants treated with sole NO₃⁻ maintained the highest survival throughout the trial, at 150% by Day 24. The control and 1:3 NH₄⁺ to NO₃⁻ fluctuated in the survival number but generally declined towards the end. Conversely, higher ammonium treatments, 1:1 and 3:1 NH₄⁺ to NO₃⁻, showed steep declines following Day 16, and sole NH₄⁺ treatment exhibited complete mortality. Overall, nitrate treatments sustained plant viability, while increasing ammonium treatments negatively influenced survival over time.
 [[File:Photos update 2.png|none|thumb|574x574px|'''Figure 8. Trial 2: Image of Plants Taken on (A) Day 0, (B) Day 7, (C) Day 14, (D) Day 21, and (E) Day 24.''' Images of plants were generally taken every Monday throughout the study duration.]]
 Algae growth was seen in 1:3 NH₄⁺ to NO₃⁻, 1:1 NH₄⁺ to NO₃⁻, and sole NH₄⁺ on Day 21. As the study progressed, treatment group photos from Day 0 to Day 24 demonstrated a decreasing sample size shown through surface area and worsening plant conditions in all groups other than 0.002M NO₃⁻ and the control.

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 '''Abstract'''</div>
-Phytoremediation is a promising method for mitigating nitrogen pollution due to its low cost and generation of biomass that can be repurposed. However, given the risk of plant invasion and the low tolerance to extreme conditions of plants currently used for phytoremediation, broad implementation requires the discovery of new species. This study assessed the phytoremediation suitability and invasive potential of ''Phyllanthus fluitans'', a promising free-floating macrophyte for which no peer-reviewed physiological data exist. ''P. fluitans'' is morphologically similar to ''Lemna minor'', the most widely used phytoremediation species, but has the added benefit of larger fronds that reduce wind-driven displacement risk. Plants were placed in 0.002M solutions of ammonium-to-nitrate ratios ranging from 0:1 to 1:0 across two independent trials. Plants receiving solely nitrate exhibited drastically higher fresh weight (p<0.05), dry weight, leaf count (p<0.05), root length, and survival rate compared to all other groups. Ammonium dose-dependently increased mortality, causing toxicity symptoms like chlorosis, necrosis, and stunted growth, resulting in complete die-off in the sole ammonium group. Additionally, the reduction of a mineral supplement between trials revealed ''P. fluitans'' to be highly sensitive to elevated water hardness. Collectively, nitrate preference and poor resilience render ''P. fluitans'' a poor phytoremediation candidate. However, under optimal conditions, the species can more than double its biomass in 24 days. This indicates that the invasive potential of ''P. fluitans'' varies drastically depending on environmental conditions, a finding that environmentalists can use to make condition-specific risk assessments and manage invasions accordingly.
+Phytoremediation is a promising method for mitigating nitrogen pollution due to its low cost and generation of biomass that can be repurposed. However, given the risk of plant invasion and the low tolerance to extreme conditions of plants fcurrently used for phytoremediation, broad implementation requires the discovery of new species. This study assessed the phytoremediation suitability and invasive potential of ''Phyllanthus fluitans'', a promising free-floating macrophyte for which no peer-reviewed physiological data exist. ''P. fluitans'' is morphologically similar to ''Lemna minor'', the most widely used phytoremediation species, but has the added benefit of larger fronds that reduce wind-driven displacement risk. Plants were placed in 0.002M solutions of ammonium-to-nitrate ratios ranging from 0:1 to 1:0 across two independent trials. Plants receiving solely nitrate exhibited drastically higher fresh weight (p<0.05), dry weight, leaf count (p<0.05), root length, and survival rate compared to all other groups. Ammonium dose-dependently increased mortality, causing toxicity symptoms like chlorosis, necrosis, and stunted growth, resulting in complete die-off in the sole ammonium group. Additionally, the reduction of a mineral supplement between trials revealed ''P. fluitans'' to be highly sensitive to elevated water hardness. Collectively, nitrate preference and poor resilience render ''P. fluitans'' a poor phytoremediation candidate. However, under optimal conditions, the species can more than double its biomass in 24 days. This indicates that the invasive potential of ''P. fluitans'' varies drastically depending on environmental conditions, a finding that environmentalists can use to make condition-specific risk assessments and manage invasions accordingly.
 ==1.0 INTRODUCTION==
@@ Line 74: / Line 74: @@
 {| style="width: 100%;border-collapse: collapse;"
 |-
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Stock Solution
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 13%;"|Stock Solution
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Amount of Distilled Water (L)
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 15%;"|Amount of Distilled Water (L)
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Concentration of Nitrate (M)
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 15%;"|Concentration of Nitrate (M)
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Concentration of Ammonium (M)
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 15%;"|Concentration of Ammonium (M)
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Mass of Ammonium chloride (g)
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 20%;"|Mass of Ammonium chloride (g)
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Mass of Potassium Nitrate (g)
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 22%;"|Mass of Potassium Nitrate (g)
 |-
 |  style="border: 1pt solid black;text-align: center;vertical-align: top;"|1
@@ Line 109: / Line 109: @@
 {| style="width: 100%;border-collapse: collapse;"
 |-
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Ratio of Ammonium to Nitrate
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 20%;"|Ratio of Ammonium to Nitrate
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Amount of solution from stock solution 1 (mL)
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 20%;"|Amount of solution from stock solution 1 (mL)
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Amount of solution from stock solution 2 (mL)
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 20%;"|Amount of solution from stock solution 2 (mL)
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Amount of solution from stock solution 3 (mL) *
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 20%;"|Amount of solution from stock solution 3 (mL) *
-|  style="border: 1pt solid black;text-align: center;vertical-align: top;"|Distilled water used to fill each tank (mL)
+|  style="border: 1pt solid black;text-align: center;vertical-align: top;width: 20%;"|Distilled water used to fill each tank (mL)
 |-
 |  style="border: 1pt solid black;text-align: center;vertical-align: top;"|* Water (control for Trial 2)

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 '''Abstract'''</div>
 Phytoremediation is a promising method for mitigating nitrogen pollution due to its low cost and generation of biomass that can be repurposed. However, given the risk of plant invasion and the low tolerance to extreme conditions of plants currently used for phytoremediation, broad implementation requires the discovery of new species. This study assessed the phytoremediation suitability and invasive potential of ''Phyllanthus fluitans'', a promising free-floating macrophyte for which no peer-reviewed physiological data exist. ''P. fluitans'' is morphologically similar to ''Lemna minor'', the most widely used phytoremediation species, but has the added benefit of larger fronds that reduce wind-driven displacement risk. Plants were placed in 0.002M solutions of ammonium-to-nitrate ratios ranging from 0:1 to 1:0 across two independent trials. Plants receiving solely nitrate exhibited drastically higher fresh weight (p<0.05), dry weight, leaf count (p<0.05), root length, and survival rate compared to all other groups. Ammonium dose-dependently increased mortality, causing toxicity symptoms like chlorosis, necrosis, and stunted growth, resulting in complete die-off in the sole ammonium group. Additionally, the reduction of a mineral supplement between trials revealed ''P. fluitans'' to be highly sensitive to elevated water hardness. Collectively, nitrate preference and poor resilience render ''P. fluitans'' a poor phytoremediation candidate. However, under optimal conditions, the species can more than double its biomass in 24 days. This indicates that the invasive potential of ''P. fluitans'' varies drastically depending on environmental conditions, a finding that environmentalists can use to make condition-specific risk assessments and manage invasions accordingly.
 ==1.0 INTRODUCTION==

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 '''2.2''' '''Preparing specimens '''
-All the plants that were used in the experiment were sourced from an online retailer and quarantined (with the same corresponding setup for the control) for 7 days to allow plants to heal from the damage of being shipped. Afterward, plants were chosen per container based on their separate examinations for health conditions regarding visible signs of nutrient deficiency, physical damage, invasion by epiphytes such as algae, possible signs of disease, etc. Trial 1 consisted of 6 samples per group, while Trial 2 consisted of 12 samples per group. All groups of plants were then gently tapped with a dry paper towel to ensure the removal of all surface liquids on the plants before measurement. Plants selected for all groups had a leaf amount of 2 to 6 leaves. The mass of the plants was taken using an analytical balance. The resulting mass was ensured to not differ more than ± 0.2 g between groups; this was done to ensure that the plants picked are made into equal groups with a similar mass.
+All the plants that were used in the experiment were sourced from an online retailer and quarantined (with the same corresponding setup for the control) for 7 days to allow plants to heal from the damage of being shipped. Afterward, plants were chosen per container based on their separate examinations for health conditions regarding visible signs of nutrient deficiency, physical damage, invasion by epiphytes such as algae, possible signs of disease, etc. In Trial 1 researchers used 6 samples per group, in account for ''P. fluitans'' reputation as an exceedingly fast growing plant. However, in Trial 2 this changed to 12 samples per group. This was done in consideration for ''P. fluitans'' delicate nature and to avoid damage occurring when handling as interpretations for treatments effects.
 '''2.3''' '''Growth Conditions'''

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 ==2 METHODOLOGY ==
-<span id='_zgy3qpwnaxgv'></span>The growth rate (and subsequent nitrogen preference, and vitality) of the plant species was observed for the duration of the experiment (28 days for Trial 1, 24 days for Trial 2). The data obtained from the investigation regarding changes in leaf growth, root growth; and wet weight was then compared to the control group (1:1 ammonium to nitrate for Trial 1, 0:0 water for Trial 2). The concentration of the nitrogen provided was based on the average nitrogen concentration in domestic wastewater due to this study’s focus on phytoremediation.
+<span id='_zgy3qpwnaxgv'></span>The growth rate (and subsequent nitrogen preference, and vitality) of the plant species was observed for the duration of the 28 days for Trial 1 and 24 days for Trial 2. The data obtained from the investigation regarding changes in leaf growth, root growth, and wet weight was then compared to the control group. A 1:1 ratio of ammonium to nitrate acted as a control for Trial 1, but was replaced by a ratio of 0:0 water for Trial 2. The concentration of the nitrogen provided in treatments was based on the average nitrogen concentration in domestic wastewater due to this study’s focus on phytoremediation.
 '''2.1''' Preparing stock solution'''
-Stock solutions were added before being poured into individual containers for experimentation. Two food-grade plastic containers with a volume of at least 2 liters, were used for making and holding the stock solutions. To organize the experimental setup, two sets of tape (with the text “stock solution 1”, “stock solution 2” and “stock solution 3 (Trial 2)” written on them) were created and stuck onto each of the 2 containers with the corresponding group name. Stock solution 1 represents the ammonium solution, stock solution 2 represents the nitrate solution, and stock solution 3 represents the water solution. Both tanks were filled with 1.9 liters of distilled water and 2.22 mL of micronutrient mix (Tropica Plant Growth Premium Fertilizer; link: [https://tropica.com/en/plant-care/liquid-fertilisers/premium-nutrition/ https://tropica.com/en/plant-care/liquid-fertilizers/premium-nutrition/]), 2.5 mL of “Seachem Flourish Phosphorus” (a potassium phosphorus-based fertilizer; link: [https://www.seachem.com/flourish-phosphorus.php https://www.seachem.com/flourish-phosphorus.php]) (removed for Trial 2), 7.0 g of “Seachem Alkaline buffer” (a commercially available sodium bicarbonate-based supplement; Link: [https://www.seachem.com/alkaline-buffer.php https://www.seachem.com/alkaline-buffer.php]), and 20 g of “Seachem equilibrium” (changed to 2.5 g for Trial 2) (a commercially available supplement/fertilizer derived from potassium sulfate, calcium sulfate, magnesium sulfate, ferric sulfate, & manganese sulfate; link: [https://www.seachem.com/equilibrium.php https://www.seachem.com/equilibrium.php]). Both batches of solution were mixed with a stirring rod to dissolve solutes and make both solutions as homogeneous as possible. In the nitrate solution (stock solution 2) 3.3 grams of potassium nitrate were added. To the ammonium solution (stock solution 1) 2.02 grams of ammonium chloride was added (concentrations displayed in Table 1).
+Stock solutions were added before being poured into individual containers for experimentation. Two food-grade plastic containers with a volume of at least 2 liters, were used for making and holding the stock solutions. To organize the experimental setup, two sets of tape (with the text “stock solution 1”, “stock solution 2” and “stock solution 3 (Trial 2)” written on them) were created and stuck onto each of the 2 containers with the corresponding group name. Stock solution 1 represented the ammonium solution, stock solution 2 represented the nitrate solution, and stock solution 3 represented the water solution. Both tanks were filled with 1.9 liters of distilled water and 2.22 mL of micronutrient mix (Tropica Plant Growth Premium Fertilizer; link: [https://tropica.com/en/plant-care/liquid-fertilisers/premium-nutrition/ https://tropica.com/en/plant-care/liquid-fertilizers/premium-nutrition/]), 2.5 mL of “Seachem Flourish Phosphorus” (a potassium phosphorus-based fertilizer; link: [https://www.seachem.com/flourish-phosphorus.php https://www.seachem.com/flourish-phosphorus.php]) (removed for Trial 2), 7.0 g of “Seachem Alkaline buffer” (a commercially available sodium bicarbonate-based supplement; Link: [https://www.seachem.com/alkaline-buffer.php https://www.seachem.com/alkaline-buffer.php]), and 20 g of “Seachem equilibrium” (changed to 2.5 g for Trial 2) (a commercially available supplement/fertilizer derived from potassium sulfate, calcium sulfate, magnesium sulfate, ferric sulfate, & manganese sulfate; link: [https://www.seachem.com/equilibrium.php https://www.seachem.com/equilibrium.php]). Both batches of solution were mixed with a stirring rod to dissolve solutes and make both solutions as homogeneous as possible. In the nitrate solution (stock solution 2) 3.3 grams of potassium nitrate were added. To the ammonium solution (stock solution 1) 2.02 grams of ammonium chloride was added (concentrations displayed in Table 1).
 After both stock solutions were made (following the concentrations shown in Table 2) they were used to create separate groups using 100% virgin plastic containers (L 175 mm x H 80 mm x W 117 mm) to test the different ratios. To distinguish between each of the tested groups each had tapes at their sides with a corresponding text stating either “0.002M nitrate”, “0.002M ammonium”, “1:3 ammonium to nitrate”, “3:1 ammonium to nitrate”, “1:1 ammonium to nitrate” (as the control for Trial 1), and “water” (as the control for Trial 2). The number of solutions from the original stock solution and distilled water that were used for these separate ratios are stated in Table 2.
@@ Line 155: / Line 155: @@
 '''2.2''' '''Preparing specimens '''
-All the plants that were used in the experiment were sourced from an online retailer and quarantined (with the same corresponding setup for the control) for 7 days to allow plants to heal from the damage of being shipped. Afterward, plants were chosen per container based on their separate examinations for health conditions regarding visible signs of nutrient deficiency, physical damage, invasion by epiphytes such as algae, possible signs of disease, etc. Trial 1 consisted of 6 samples per group, while Trial 2 consisted of 12 samples per group. All groups of plants were then gently tapped with a dry paper towel to ensure the removal of all surface liquids on the plants before measurement. Plants selected for all groups had a leaf amount of 2 to 6 leaves. The mass of the plants was taken using an analytical balance. The resulting mass should not differ more than ± 0.2 g between groups; this was done to ensure that the plants picked are made into equal groups with a similar mass.
+All the plants that were used in the experiment were sourced from an online retailer and quarantined (with the same corresponding setup for the control) for 7 days to allow plants to heal from the damage of being shipped. Afterward, plants were chosen per container based on their separate examinations for health conditions regarding visible signs of nutrient deficiency, physical damage, invasion by epiphytes such as algae, possible signs of disease, etc. Trial 1 consisted of 6 samples per group, while Trial 2 consisted of 12 samples per group. All groups of plants were then gently tapped with a dry paper towel to ensure the removal of all surface liquids on the plants before measurement. Plants selected for all groups had a leaf amount of 2 to 6 leaves. The mass of the plants was taken using an analytical balance. The resulting mass was ensured to not differ more than ± 0.2 g between groups; this was done to ensure that the plants picked are made into equal groups with a similar mass.
 '''2.3''' '''Growth Conditions'''
@@ Line 163: / Line 163: @@
 '''2.4''' '''Maintaining plants/experimenting'''
-Trial 1’s sample size was 6 samples per group, while Trial 2’s sample size was 12 samples per group. After being measured, plants with 2 to 6 leaves were gently placed on the water's surface of each of the groups in their corresponding separate containers. Throughout the experiment, all groups were provided with 10 hours of light per day for 28 days while placed near one another to prevent inconsistencies in temperature for Trial 1. The same measures were taken for Trial 2, but in a 24-day duration. The plant medium/solution was replaced every Monday to maintain proper levels of nutrients with the same process repeated on maintained on the prepared pots and conditions. Plants were periodically removed and wrapped with a paper towel soaked in distilled water while this procedure was occurring. Throughout this procedure, all unexpected occurrences (deteriorating plant health, abnormal amount of dead plant matter, chlorosis, etc.) were recorded.
+The sample size for Trial 1 was 6 samples per group, while Trial 2 had a sample size of 12 per group. After being measured, plants with 2 to 6 leaves were gently placed on the water's surface of each of the groups in their corresponding separate containers. Throughout the experiment, all groups were provided with 10 hours of light per day for 28 days while placed near one another to prevent inconsistencies in temperature for Trial 1. The same measures were taken for Trial 2, but in a 24-day duration. The plant medium/solution was replaced every Monday to maintain proper levels of nutrients with the same process repeated on maintained on the prepared pots and conditions. Plants were periodically removed and wrapped with a paper towel soaked in distilled water while this procedure was occurring. Throughout this procedure, all unexpected occurrences (deteriorating plant health, abnormal amount of dead plant matter, chlorosis, etc.) were recorded.
 '''2.5''' '''Measuring Root Length & Leaf Numbers'''

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 In Trial 1, Fig. 4a & Fig. 4b, total leaf amount generally declined with combined nitrate and ammonium treatments. By Day 28, 0.002M NO₃⁻ showed the greatest increase in total leaf count relative to other treatments (45) followed by 0.002M NH₄⁺ (17). In contrast, combined treatments (1:3, 1:1, and 3:1 NH₄⁺ to NO₃⁻) showed reductions in leaf number over time (1, 4, and 0 by Day 28).In Trial 2, Fig. 4c & Fig. 4d, total leaf amount increased over time, primarily in the 0.002M NO₃⁻ group, showing a statistically significant increase from Day 0 (42) to Day 24 (140), and had the highest overall leaf number. The control group showed a moderate increase by Day 14, while the 1:3 NH₄⁺ to NO₃⁻ group showed some growth at Day 14 (77), then decreased by nearly 50% at Day 24 (38). Conversely, the 1:1 and 3:1 NH₄⁺ to NO₃⁻ treatments significantly reduced total leaf amount over time, and the 0.002M NH₄⁺ consistently had the lowest leaf count.
 [[File:Survival percentage update.png|none|thumb|600x600px|'''Figure 6. Survival Percentage with Time.''' (a) Trial  1: the sample size is a percentage from Day 0 to Day 28. (b) Trial 2: the sample size is a percentage from Day 0 to Day 24]]
-Similar to Figures 1-5, survival rates depended on the nitrogen sources over time. In Trial 1, Fig. 6a, survival generally remained stable in the 0.0022M NO₃⁻ treatment, maintaining 100% survival by Day 28. The 0.002M NH₄⁺ treatment group initially increased to 116.67% and remained stable through Day 11, but declined to 50% of the original sample size at Day 14. Conversely, mixed nitrogen sources showed a reduction in survival percentage over time, with 3:1 NH₄⁺ to NO₃⁻ treatment reaching complete mortality by Day 28. In Trial 2, Fig. 6b, plants treated with sole NO₃⁻ maintained the highest survival throughout the trial, at 150% by Day 24. The control and 1:3 NH₄⁺ to NO₃⁻ fluctuated in the survival number but generally declined towards the end. Conversely, higher ammonium treatments, 1:1 and 3:1 NH₄⁺ to NO₃⁻, showed steep declines following Day 16, and sole NH₄⁺ treatment exhibited complete mortality. Overall, nitrate treatments sustained plant viability, while increasing ammonium treatments negatively influenced survival over time.<div class="center" style="width: auto; margin-left: auto; margin-right: auto;">
+Similar to Figures 1-5, survival rates depended on the nitrogen sources over time. In Trial 1, Fig. 6a, survival generally remained stable in the 0.0022M NO₃⁻ treatment, maintaining 100% survival by Day 28. The 0.002M NH₄⁺ treatment group initially increased to 116.67% and remained stable through Day 11, but declined to 50% of the original sample size at Day 14. Conversely, mixed nitrogen sources showed a reduction in survival percentage over time, with 3:1 NH₄⁺ to NO₃⁻ treatment reaching complete mortality by Day 28. In Trial 2, Fig. 6b, plants treated with sole NO₃⁻ maintained the highest survival throughout the trial, at 150% by Day 24. The control and 1:3 NH₄⁺ to NO₃⁻ fluctuated in the survival number but generally declined towards the end. Conversely, higher ammonium treatments, 1:1 and 3:1 NH₄⁺ to NO₃⁻, showed steep declines following Day 16, and sole NH₄⁺ treatment exhibited complete mortality. Overall, nitrate treatments sustained plant viability, while increasing ammonium treatments negatively influenced survival over time.<div class="center" style="width: auto; margin-left: auto; margin-right: auto;">[[File:Images update.png|none|thumb|589x589px|'''Figure 7. Trial 1: Image of Plants Taken on (a) Day 0, (b) Day 7, (c) Day 14, (d) Day 21, and (e) Day 28.''' Images of plants were generally taken every Monday throughout the study duration.]]</div>Algae growth was seen in 0.002M NO₃⁻, 1:1 NH₄⁺ to NO₃⁻, and 3:1 NH₄⁺ to NO₃⁻. Comparing treatment group photos from Day 0 (A) to Day 28 (E), sample sizes decreased as plant conditions worsened.
+[[File:Photos update 2.png|none|thumb|574x574px|'''Figure 8. Trial 2: Image of Plants Taken on (A) Day 0, (B) Day 7, (C) Day 14, (D) Day 21, and (E) Day 24.''' Images of plants were generally taken every Monday throughout the study duration.]]
-[[Image:Draft_Zhang_868588273-picture-Group 13.svg|center|600px]]'''Figure 5. Image of Plants Taken on (A) Day 0, (B) Day 7, (C) Day 14, (D) Day 21, and (E) Day 24.''' Images of plants were generally taken every Monday throughout the study duration.
+Algae growth was seen in 1:3 NH₄⁺ to NO₃⁻, 1:1 NH₄⁺ to NO₃⁻, and sole NH₄⁺ on Day 21. As the study progressed, treatment group photos from Day 0 to Day 24 demonstrated a decreasing sample size shown through surface area and worsening plant conditions in all groups other than 0.002M NO₃⁻ and the control.
-−
-Algae growth was seen in 1:3 NH₄⁺ to NO₃⁻, 1:1 NH₄⁺ to NO₃⁻, and sole NH₄⁺ on Day 21. As the study progressed, treatment group photos from Day 0 to Day 24 demonstrated a decreasing sample size shown through surface area and worsening plant conditions in all groups other than 0.002M NO₃⁻ and the control.[[File:Images update.png|none|thumb|589x589px|'''Figure 7. Trial 1: Image of Plants Taken on (a) Day 0, (b) Day 7, (c) Day 14, (d) Day 21, and (e) Day 28.''' Images of plants were generally taken every Monday throughout the study duration.]]</div>Algae growth was seen in 0.002M NO₃⁻, 1:1 NH₄⁺ to NO₃⁻, and 3:1 NH₄⁺ to NO₃⁻. Comparing treatment group photos from Day 0 (A) to Day 28 (E), sample sizes decreased as plant conditions worsened.
 ==5.0 DISCUSSION==

@@ Line 196: / Line 196: @@
 [[Image:Draft_Zhang_868588273-picture-Group 13.svg|center|600px]]'''Figure 5. Image of Plants Taken on (A) Day 0, (B) Day 7, (C) Day 14, (D) Day 21, and (E) Day 24.''' Images of plants were generally taken every Monday throughout the study duration.
-Algae growth was seen in 1:3 NH₄⁺ to NO₃⁻, 1:1 NH₄⁺ to NO₃⁻, and sole NH₄⁺ on Day 21. As the study progressed, treatment group photos from Day 0 to Day 24 demonstrated a decreasing sample size shown through surface area and worsening plant conditions in all groups other than 0.002M NO₃⁻ and the control.
+Algae growth was seen in 1:3 NH₄⁺ to NO₃⁻, 1:1 NH₄⁺ to NO₃⁻, and sole NH₄⁺ on Day 21. As the study progressed, treatment group photos from Day 0 to Day 24 demonstrated a decreasing sample size shown through surface area and worsening plant conditions in all groups other than 0.002M NO₃⁻ and the control.[[Image:Draft_Zhang_868588273-picture- 1.svg|center|600px|'''Figure 7. Trial 1: Image of Plants Taken on (A) Day 0, (B) Day 7, (C) Day 14, (D) Day 21, and (E) Day 28.''' Images of plants were generally taken every Monday throughout the study duration.]]</div>Algae growth was seen in 0.002M NO₃⁻, 1:1 NH₄⁺ to NO₃⁻, and 3:1 NH₄⁺ to NO₃⁻. Comparing treatment group photos from Day 0 (A) to Day 28 (E), sample sizes decreased as plant conditions worsened.<div class="center" style="width: auto; margin-left: auto; margin-right: auto;">
 </div>
-<div class="center" style="width: auto; margin-left: auto; margin-right: auto;">
+[[Image:Draft_Zhang_868588273-picture- 1.svg|center|600px]]'''Figure 7. Trial 1: Image of Plants Taken on (A) Day 0, (B) Day 7, (C) Day 14, (D) Day 21, and (E) Day 28.''' Images of plants were generally taken every Monday throughout the study duration.
- [[Image:Draft_Zhang_868588273-image25.png|390px]] </div>'''Figure 6. Trial 2 Survival Percentage with Time.''' The sample size is a percentage from Day 0 to Day 24.
-−
-Similar to Figures 1-5, survival rates depended on the nitrogen sources over time. Plants treated with sole NO₃⁻ maintained the highest survival throughout the trial, at 150% by Day 24. The control and 1:3 NH₄⁺ to NO₃⁻ fluctuated in the survival number but generally declined towards the end. Conversely, higher ammonium treatments, 1:1 and 3:1 NH₄⁺ to NO₃⁻, showed steep declines following Day 16, and sole NH₄⁺ treatment exhibited complete mortality. Overall, nitrate treatments sustained plant viability, while increasing ammonium treatments negatively influenced survival over time. <div class="center" style="width: auto; margin-left: auto; margin-right: auto;">[[Image:Draft_Zhang_868588273-picture-Group 12.svg|center|600px]]'''Figure 7. Trial 1 Total Fresh Weight with Time.''' (A) As a progression from Day 0 to Day 28 and (B) Day 0, Day 14, and Day 24 on individual dates. Error bar values represent mean ±STDEV. Statistical significance was not calculated due to insufficient sample sizes in some treatment groups.
-−
-In Fig. 7, total fresh weight generally decreased over time across most treatment groups, with the exception of the 0.002M NO₃⁻ treatment (0.3499 g by Day 28). Ammonium-containing treatments showed a substantial decline through Day 28.
-</div>
-−
-<div class="center" style="width: auto; margin-left: auto; margin-right: auto;">
- [[Image:Draft_Zhang_868588273-image30.png|324px]] </div>'''Figure 8. Trial 2 Dry Weight Per Treatment Group.''' The end samples per treatment varied: 0.002 M NO₃⁻ (6 samples), 1:3 (1 sample), 1:1 (1 sample), 3:1 (0 samples), and 0.002 M NH₄⁺ (3 samples).
-−
-Final dry weight differed noticeably across nitrogen treatments as a result of varying sample sizes. The 0.002M NO₃⁻ had the highest overall dry weight (0.0429 g), followed by the 0.002M NH₄⁺ treatment (0.0309 g). Treatments containing both nitrate and ammonium yielded considerably lower dry mass. The 3:1 NH₄⁺ to NO₃⁻ treatment group had no remaining samples at Day 28, preventing measurement of dry weight.
-[[Image:Draft_Zhang_868588273-picture-Group 14.svg|center|600px]]'''Figure 9. Mean Root Length with Time'''. (A) As a progression from Day 0 to Day 28 and (B) Day 0, Day 14, and Day 28 on individual dates. Error bar values represent mean ±STDEV. Statistical significance was not calculated due to insufficient sample sizes in some treatment groups.
-−
-In Fig. 9, mean root length generally decreased over time across all nitrogen treatments. The 0.002M NO₃⁻ group had the highest average root length by Day 28 compared to other treatments. <div class="center" style="width: auto; margin-left: auto; margin-right: auto;">
-−
-[[Image:Draft_Zhang_868588273-picture-Group 15.svg|center|600px]]'''Figure 10. Total Leaf Amount with Time'''. (A) As a progression from Day 0 to Day 28 and (B) Day 0, Day 14, and Day 28 on individual dates. Error bar values represent mean ±STDEV. Statistical significance was not calculated due to insufficient sample sizes in some treatment groups.
-−
-Total leaf amount generally declined with combined nitrate and ammonium treatments. By Day 28, 0.002M NO₃⁻ showed the greatest increase in total leaf count relative to other treatments (45) followed by 0.002M NH₄⁺ (17). In contrast, combined treatments (1:3, 1:1, and 3:1 NH₄⁺ to NO₃⁻) showed reductions in leaf number over time (1, 4, and 0 by Day 28).
-</div>
-−
-<div class="center" style="width: auto; margin-left: auto; margin-right: auto;">
-−
-[[Image:Draft_Zhang_868588273-picture- 1.svg|center|600px]]'''Figure 11. Image of Plants Taken on (A) Day 0, (B) Day 7, (C) Day 14, (D) Day 21, and (E) Day 28.''' Images of plants were generally taken every Monday throughout the study duration.
 Algae growth was seen in 0.002M NO₃⁻, 1:1 NH₄⁺ to NO₃⁻, and 3:1 NH₄⁺ to NO₃⁻. Comparing treatment group photos from Day 0 (A) to Day 28 (E), sample sizes decreased as plant conditions worsened.
 </div>
 ==5.0 DISCUSSION==