Review Reports

Response to Reviewer 1 Comments

1. Summary

We sincerely thank the reviewer and the editor for their thorough evaluation and constructive comments on our manuscript. The suggestions provided were highly valuable and have significantly improved the clarity, accuracy, and overall quality of the paper. All comments have been carefully considered, and the corresponding revisions have been incorporated into the revised version of the manuscript. All changes in the revised manuscript are highlighted in green for easy reference.

2. Questions for General Evaluation

Reviewer’s Evaluation

Response and Revisions

Does the introduction provide sufficient background and include all relevant references?

Yes

Are all the cited references relevant to the research?

Yes

Is the research design appropriate?

Yes

Are the methods adequately described?

Can be improved

Are the results clearly presented?

Can be improved

Are the conclusions supported by the results?

Yes

3. Point-by-point response to Comments and Suggestions for Authors

Comments 1: Line 114: what did 4 dm refer to? Diameter or radius?

Response 1: Dear Reviewer, Thank you for your comments. The expression “4 dm³” refers to a volume measurement (cubic decimeters), not to a linear dimension such as diameter or radius. To ensure consistency with the International System of Units (SI), this unit will be converted to cubic meters (m³) in the revised version of the manuscript.

Comments 2: Lines 125-127: These parameters are typically reported as mg/kg and cmolc/kg as per weight of soil instead of volume since extraction volume can be different among studies. Please convert this unit to dry weight of soil.

Response 2: Dear reviewer, thank you for this valuable observation. As suggested, the parameters previously expressed on a volumetric basis (mg dm⁻³ and cmolc dm⁻³) have been converted to the corresponding units on a dry weight basis (mg kg⁻¹ and cmolc kg⁻¹) to ensure consistency with standard soil analysis conventions and comparability among studies. The revised table now presents the soil chemical properties as mg kg⁻¹ for nutrient concentrations and cmolc kg⁻¹ for exchangeable cations.

Comments 3: Line 166: please provide information about the chemical fertilizer, such as composition, apply rate, etc.

Response 3: Thank you for the observation. Information regarding the composition and amount of chemical fertilizers used has been added to the revised manuscript. Soil analyses were carried out prior to the experiment to assess fertility and determine the nutrient correction requirements. Based on these results, a single soil application of chemical fertilizer was performed at the beginning of the experiment, following regional agronomic recommendations (Embrapa Amazônia Ocidental, 2020).

Comments 4: Line 172: 384 ml? Why? Please add justification of the amount of nutrients added.

Response 4: We appreciate the reviewer’s valuable comment. The daily irrigation volume of 384 mL was initially based on the methodology proposed by Klar (1966), which recommends maintaining soil moisture at approximately 60% of field capacity. However, in the revised version of the manuscript, the text will be adjusted to clarify that the irrigation volume will be adjusted to meet the specific water demand of the plants, ensuring adequate soil moisture conditions for optimal growth. This approach better reflects the dynamic nature of plant water requirements throughout the experimental period.

Comments 5: Line 184-185: daily water samples were collected. But it was not reported in the results. Also, it conflicted with line 194 of weekly monitoring of nitrogenous compounds. This is confusing. Please clarify this and revise it to be consistent.

Response 5: We thank the reviewer for pointing out this inconsistency. To clarify, daily water samples were collected to monitor general water quality parameters such as temperature, pH, dissolved oxygen, and electrical conductivity. In contrast, nitrogenous compounds (ammonia, nitrite, and nitrate) were analyzed on a weekly basis. We have revised the manuscript to clearly reflect this distinction and ensure consistency.

Comments 6: Line 198: Each sample? It’s not clear how many samples were collected for each treatment. Please add this information.

Response 6: We appreciate the reviewer’s observation. Water samples for the analysis of nitrogenous compounds were collected once per week from each treatment. Therefore, the expression “each sample” refers to the weekly sample collected from each treatment unit throughout the experimental period. This information has been added to the revised version of the manuscript for clarity.

Comments 7: Line 212-227: Please add fish density/population in this session. The authors had some
information in the discussion, please move it here.

Response 2: We thank the reviewer for this helpful comment. The information regarding the fish population has been added to this section, as suggested. A total of 51 tambaqui (C. macropomum) were stocked in the recirculating aquaculture system, with a combined biomass of 25.295 kg, corresponding to the grow-out phase of the species.

Comments 8: Line 229-248: authors need to add more sampling information to this part. a. Please justify the sampling schedule in this study. It seems Jambu takes longer than 50 days as the last harvest in this study, so 50 days is not enough time to represent the fertilizer needs and growth trend. b. How/how many samples were collected for each treatment/each pot?

Response 8: (a) We appreciate the reviewer’s observation. The 50-day cultivation period was defined based on the commercial harvest stage of A. oleracea (jambu) in the Amazon region, where the crop is commonly harvested between 45 and 60 days after sowing, depending on environmental and management conditions. In this study, the goal was to assess the early to intermediate growth response of jambu to aquaculture effluent irrigation, rather than to reach the full physiological maturity of the plants. Thus, the 50-day period adequately represented the stage at which the species shows significant vegetative growth and is typically harvested for culinary and commercial purposes.

b) We appreciate the reviewer’s question. At the end of the experimental period, six plants were collected per treatment for phytometric and biomass analyses. The plants were carefully removed from the cultivation pots, washed to eliminate residual substrate, and subsequently evaluated for morphological parameters. This procedure ensured the accuracy and representativeness of the data used to assess the growth performance of A. oleracea under the different irrigation treatments.

Comments 9: Line 269-272: Since authors mentioned that daily water samples were collected and analyzed, so why not a line chart to show the trend? The average didn’t quite catch all the variations,
especially for total dissolved solids with such a big range (189-758), which related to nutrients content in the effluent.

Response 9: We appreciate the reviewer’s valuable suggestion. A line chart will be included in the revised version of the manuscript to better illustrate the temporal variations in the water quality parameters, particularly total dissolved solids, whose fluctuations reflect the nutrient dynamics in the effluent. This addition will provide a clearer representation of the trends observed throughout the experimental period.

Comments 10: Line 285: caption for a figure should be put below the figure instead of above.

Response 10: We thank the reviewer for pointing this out. The figure captions have been moved below the figures in accordance with the journal’s formatting guidelines.

Comments 11: Line 287: please explain x20 and x40 in this figure.

Response 11: We appreciate the reviewer’s observation. A correction factor was not initially applied when generating the graph. The values were subsequently updated after applying the titration correction factor, and the graph was regenerated to display the corrected concentrations of these variables. The indications “×20” and “×40” have been removed, as they referred to the dilution factors used during the laboratory titration process and were not meant to appear in the final figure. The updated version now accurately represents the corrected values.

Comments 12: Line 287: the harvest was after 50 days. Why does the data here only show about 30 days? So, no more analysis after 30 days?

Response 12: Thank you for your observation. The experimental period was considered 50 days because it included both the seed germination phase (which lasted approximately 25 days) and the subsequent treatment period (around 25 days). The plants were subjected to the treatments only after germination was completed. Therefore, the total cultivation time was 50 days, considering the germination phase as part of the experimental period.

Comments 13: Line 290-299: some results here are redundant. Please concise.

Response 13: We thank the reviewer for the suggestion. The Results section has been revised to remove redundancy and present the main findings more concisely, while preserving the essential statistical comparisons and treatment effects.

Comments 14: Line 304-305: table 2: only SD had a significant difference among 100%, 75%, and 50% fertilizer. There were no significant differences among all other growth parameters even though a reduction were observed in 75% and 50% treatment.

Response 14: We thank the reviewer for the careful observation. The text has been revised to accurately reflect the statistical outcomes, indicating that only stem diameter (SD) showed significant differences among the 100%, 75%, and 50% fertilizer treatments, while other growth parameters did not differ significantly despite slight reductions in the 75% and 50% fertilizer treatments.

Comments 15: Line 304-305: DRW had a pretty big variation among each treatment. Please explain this in the discussion.

Response 15: We appreciate the observation regarding the relatively high variation in DRW among treatments. This variability most likely reflects the natural heterogeneity in root biomass accumulation in A. oleracea under different fertigation regimes. As noted by Nendel et al. (2009), root growth in vegetable crops is highly sensitive to localized fluctuations in nutrient availability and soil moisture, which can result in considerable within-treatment variation even under controlled conditions. In addition, A. oleracea possesses a shallow and fibrous root system, which tends to respond rapidly to microenvironmental differences, thereby amplifying variability in dry root weight. We have included a clarification in the Discussion section to explain this point.

Comments 16: Line 311: 800 L or 8000 L in line 130?

Response 16: We appreciate the reviewer’s careful observation. There was a typographical error in the manuscript — the correct value is 8000 L, as now corrected in line 130.

Comments 17: Line 318: 399.80 g? Seems an off number.

Response 17: Thank you for your comment. The initial mean weight of the fish was 228.57 g, and after approximately one month the individuals reached an average weight of 399.80 g, representing an increase of 171.23 g (about 5.7 g day⁻¹ or 74.9% relative gain). This result indicates a satisfactory monthly growth rate of Colossoma macropomum under the experimental conditions.

Comments 18: The authors didn’t have a full nutrient analysis. How did authors think about other nutrient concentration? Any other low nutrients that inhibit limitations? Please add discussion per this

Response 18: Thank you for your observation. Although a complete nutrient analysis was not performed, the evaluation of plant nutritional status was based on visual and morphological indicators, which are reliable measures of nutrient sufficiency in Acmella oleracea. Throughout the experiment, plants exhibited uniform growth, intense green coloration, and no visible deficiency symptoms such as chlorosis or necrosis. These observations suggest that essential nutrients were available in sufficient amounts to support normal metabolism and biomass accumulation. Therefore, while we acknowledge this as a methodological limitation, the overall health and productivity of the plants indicate that nutrient availability was not a limiting factor in this study.

Comments 19: Line 344: add reference.

Response 19: Thank you for the observation. The appropriate reference has been added in the revised manuscript to support this statement.

Comments 20: Line 344: 30 days or 50 days?

Response 20: Thank you for your comment. The total duration of the experiment was 50 days, which included approximately 25 days for seed germination and 30 days for the treatment period. Therefore, while the cultivation cycle lasted 50 days in total, the experimental phase under the applied treatments covered 30 days.

4. Response to Comments on the Quality of English Language

Point 1: The English is fine and does not require any improvement.

Response 1: The English is fine and does not require any improvement.

5. Additional clarifications

Response to Reviewer 2 Comments

1. Summary

We sincerely thank the reviewer and the editor for their thorough evaluation and constructive comments on our manuscript. The suggestions provided were highly valuable and have significantly improved the clarity, accuracy, and overall quality of the paper. All comments have been carefully considered, and the corresponding revisions have been incorporated into the revised version of the manuscript. All changes in the revised manuscript are highlighted in blue for easy reference.

2. Questions for General Evaluation

Reviewer’s Evaluation

Response and Revisions

Does the introduction provide sufficient background and include all relevant references?

Can be improved

Are all the cited references relevant to the research?

Can be improved

Is the research design appropriate?

Can be improved

Are the methods adequately described?

Can be improved

Are the results clearly presented?

Can be improved

Are the conclusions supported by the results?

Can be improved

3. Point-by-point response to Comments and Suggestions for Authors

Comments 1: The title - problem with the word "bioeconomic"

Response 1: Thank you for this observation. We agree that the term bioeconomic may lead to a misinterpretation of the study’s scope, as our research focuses primarily on the biological performance of plants and fish in an integrated aquaculture-agriculture system rather than on economic analysis. Accordingly, the title has been revised to better reflect the biological nature of the study.

Comments 2: Line 103 - Correct: The region has a humid equatorial climate (Köppen classification: Af), characterized by consistently high temperatures and high relative humidity throughout the year.

Response 2: Thank you for the suggestion. The sentence has been corrected in the revised manuscript as follows: “The region has a humid equatorial climate (Köppen classification: Af), characterized by consistently high temperatures and high relative humidity throughout the year.”

Comments 3: Table 1 - dm^-3, please convert to L^-1.

Response 3:

Comments 4: Line 134 - Provide power in kW.  and L/min

Response 4: Thank you for the observation. The information regarding the blower power and air flow rate has been added in the revised version of the manuscript.

Comments 5: Figure 1 - The graphic description should be below the graphic.

Response 5: Thank you for the observation. The figure layout has been revised, and the graphic description (caption) is now placed below the figure, in accordance with the journal’s formatting guidelines.

Comments 6: Line 179 - The word “factorial” implies that the influence of at least two factors was studied. Correct the sentence to: A single-factor design with multiple treatment levels was used to evaluate the effect of chemical and biological nutrient sources on plant biomass productivity and nutrient use efficiency.

Response 6: Thank you for the observation. The sentence has been corrected in accordance with the suggestion to accurately describe the experimental design.

Comments 7: Line 222 - Were these individuals labeled? How many were there, and is the number given here an average value? What was the average?

Response 7: Thank you for the valuable comment. The information regarding the number and initial average weight of the fish has been clarified in the revised manuscript. The individuals were not individually labeled; however, the total number of fish and their mean initial weight have now been included to provide full transparency.

Comments 8: Line 266,267 - Probably one significant digit too precise.

Response 8: Thank you for the observation. The values have been rounded to one decimal place to better reflect the precision of the measurements. 

Comments 9: Figure 2 - The figure description should be below the graphic. Bold text in the graphic should be larger.

Response 9: Thank you for the suggestion. The figure layout has been revised accordingly — the description (caption) is now placed below the graphic, and the bold text within the figure has been enlarged to improve readability and visual balance.

Comments 10: Line 302 - A. oleracea - write in italics.  A. oleracea

Response 10: We thank the reviewer for pointing this out. The figure captions have been moved below the figures in accordance with the journal’s formatting guidelines.

Comments 11: Line 303 - correct to "Numbers followed by different letters indicate statistically significant differences between densities (p  ≤ 0.05)"

Response 11: Thank you for the observation. The sentence has been corrected in the revised manuscript as follows: “Numbers followed by different letters indicate statistically significant differences between densities (p ≤ 0.05).”

Comments 12: Line 313 - the word "initial" was used - the numbers indicating the fish weight seem to be incorrect - 51 fish weighed approximately 25.3 kg, so the average weight was approximately 0.5 kg. The word "initial" was used in line 315, but the average weight was lower, 0.2286 kg. Please write this more clearly - preferably in order - first the data at the beginning of the experiment, then the data at the end of the experiment.

Response 12: Thank you for the observation. We appreciate the reviewer’s attention to detail. The paragraph has been revised to clarify the sequence of events and to ensure consistency between the reported values for the beginning and end of the experiment. The confusion occurred due to the placement of “initial” and the order of presentation

Comments 13: Line 349 - missing Ferreira da Costa et al. in References

Response 13: Thank you for the observation. The missing reference (Ferreira da Costa et al.) has been added to the References section in the revised manuscript.

4. Response to Comments on the Quality of English Language

Point 1: English is fine.  Needs more precision in some places.

Response 1: We appreciate the reviewer’s observation. A thorough language revision was performed, and all issues requiring greater precision have been addressed.

5. Additional clarifications