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PONE-D-25-68057

Nutritional Value of Hora and Mineral-enriched Soil for Cattle: Evaluation of Mineral Composition and Physicochemical Properties in Southwestern Ethiopia.

PLOS One

Dear Dr. Miresa,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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ACADEMIC EDITOR:

Changes  required for acceptance:

- Expand the discussion on the absence of control samples (areas distant from the “hora” sources), highlighting how this limitation may influence the interpretation of the degree of mineral enrichment.

- Include a paragraph addressing the potential impact of seasonality (especially the rainy season) on mineral concentrations, total dissolved solids, and animal intake.

- Explicitly emphasize that the estimated mineral intake values represent gross intake rather than bioavailable intake, considering: (1) the alkaline soil pH, which may reduce absorption; and (2) possible antagonistic interactions among minerals (e.g., Fe, Mo, and Mn affecting copper metabolism).

- Provide additional information on the selenium analysis by HG-ICP-OES, particularly limits of quantification (LOQ) and/or recovery rates, to strengthen analytical transparency and robustness.

- Conduct a general grammatical revision of the manuscript.

- Standardize citations and references, correcting the numbering throughout the text (e.g., adjust “CCME (2025)” to the appropriate format, such as “CCME [27]”).

- Review punctuation and formatting in the indicated lines (p. 5, lines 129–130).

- Correctly format chemical notation (e.g., use subscript in NH₃).

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Academic Editor

PLOS One

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: General Evaluation

This study provides a valuable quantitative assessment of a traditional livestock feeding practice by evaluating the mineral composition and physicochemical properties of "hora" (natural mineral water) and its surrounding mineral-enriched soils in southwestern Ethiopia. By analyzing samples from the Bedele, Dabo, Gechi, and Borecha districts, the authors effectively highlight the potential of these resources as a strategic natural source of sodium and selenium, while also correctly identifying the toxicological risks associated with excessive iron, molybdenum, and manganese in the soil.

The methodology, incorporating APHA standards for water and ICP-OES analysis for mineral quantification, is technically sound. Furthermore, modeling the potential daily mineral intake based on established water and dry matter consumption formulas for tropical cattle is a highly commendable approach that bridges analytical chemistry with practical animal nutrition. The manuscript aligns well with the publication criteria of PLOS ONE, presenting original research with rigorously supported conclusions. However, addressing a few methodological limitations and contextual factors in the discussion will significantly strengthen the final manuscript.

Specific Comments

Baseline and Control Data: As the authors rightly acknowledged in the discussion, the study currently lacks a comparative baseline from sites distal to the hora springs. Without these control samples for background water and soil, quantifying the exact magnitude of mineral enrichment relative to the region’s basal environmental status remains difficult. I recommend expanding the discussion slightly to address how this limitation might influence the interpretation of the net dietary mineral balance.

Seasonal Variability: The sampling was exclusively conducted during the dry season (December-February). It is highly probable that precipitation during the wet season would dilute the total dissolved solids (TDS) and alter specific mineral concentrations. Please add a brief paragraph discussing the potential impact of seasonal dynamics on water palatability, mineral concentration, and consequently, the variations in the cattle’s daily mineral intake throughout the year.

Bioavailability vs. Gross Intake: The daily soil ingestion was reasonably estimated at 0.5 kg DM/hd/day. While the gross mineral intake calculations presented in Table 4 are helpful, the alkaline pH recorded in the soil samples (e.g., reaching up to 8.1 in Bedele and 8.28 in Dabo) does not favor the optimal absorption of most minerals in the gastrointestinal tract. Furthermore, the extremely high levels of iron, molybdenum, and manganese could induce secondary copper deficiency due to strong antagonistic interactions. I suggest explicitly emphasizing that the calculated gross intake does not equal the bioavailable intake, particularly under these alkaline and highly antagonistic matrix conditions.

Methodological Clarity (HG-ICP-OES): The use of microwave digestion and subsequent hydride generation (HG-ICP-OES) for selenium determination is an excellent methodological choice to enhance sensitivity and minimize matrix interferences. To further validate this analytical rigor, please consider adding a brief statement in the methodology section regarding the recovery rates or the limits of quantification (LOQ) specific to this selenium analysis.

Reviewer #2: Editor

I appreciate the trust placed in me to review the article titled: “Nutritional Value of Alfalfa and Mineral-Enriched Soil for Livestock: Evaluation of Mineral Composition and Physicochemical Properties in Southwest Ethiopia.” The article is well-structured, and the study’s objective is relevant.

Some minor grammatical details have been identified. Regarding the citation of references within the document, a careful review is necessary, and the numbering should be restructured. The inconsistencies begin at the foot of Table 1, where it appears as: CCME (2025) and should be CCME [27].

Other grammatical details are observed on page 5, lines 129, 130 Please review the punctuation and subscript in the chemical formula NH3, line 132.

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Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: PLOS ONE-Reviewer Report.doc

Nutritional value of hora and mineral-enriched soil for cattle: Evaluation of mineral composition and physicochemical properties in southwestern Ethiopia.

PONE-D-25-68057R1

Dear Dr. Miresa,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Elma Izze Da Silva Magalhães

Academic Editor

PLOS One