Impact of Effective Microorganisms (EM) Application on the Physical Condition of Haplic Luvisol
Round 1
Reviewer 1 Report
The paper “Impacts of effective microorganisms (EM) application on the physical conditions of Haplic Luvisol” , analyzes the impact of effective microorganisms application on soil physical properties in a field essay along five years. This essay complement a previous one that analyses the short term effect of EM application on soil properties. In this work the EM application is annualy repeated in a crop rotation essay along 5 years. In each one of te analized years the soil management was different as dictated by crop rotation and for this reason the year is considered as an additional variability factor.
Comments
Introduction
As EM is mainly employed to increase the soil biodiversity in order to stabilize the ecosystem, some hypothesis must be stablished to justify why the authors repeat the treatment several years.
Material and methods
Analysis section:
Apparently the two kind of simples were taken, undisturbed samples for water retention and porosity and disturbed samples used to analyze water stability of aggregates. Did all the other physical analysis realize over the same sample ¿. In this case, what is the sequence of the samples treatment?
Regarding the statistical methods, the authors employ two way ANOVA but the results are not congruents with this analysis. The two way ANOVA considers two independent factors, treatment as main factor and time as secondary factor. LSD (Least Significant Difference) is the value at a particular level of statistical probability when exceeded by the difference between two varietal means for a particular characteristic, then the two varieties are said to be distinct for that characteristic.The hypothesis are that the factor applicaton of EM or the factor year of analysis produced different mean values. ANOVA result are not available since the tables have not been added to the manuscript.
Results and discussion
The results can not be correctly assesed because tables are not been included in the manuscript. But it seems better reorder the section, analyzing first the effect of EM application on soil properties and using a different section to discuss correlations
Minor comments:
Line 12: Is preparatu used correctly in english?
Line 23: What does mean “ requires serious consideration”?
Lines 36-37: Can you reformulate the paragraph: They are……..that are
Line 42-43: Soil structure is highly dependent on soil texture
Lines 66-69 Probably the efficiency of EM application under field condition is related to the microbiological diversity and quality of the soil. Moreover the application of microorganisms must be complemented with the application of substrate in order to stabilize the microbial population
Line 97: does the compaction and the hydraulic conductivity decrease simultaneously?
Lines 151-155: It is recomended move this paragraph to the begining of the section.
Lines 137- 150. Please, describe first the crop rotation and after the EM application
Line 163: Does it mean one replicate in each plot? The physical properties are highly variables and dependent on soil conditions. Why did you perform three sampling data per year? Probably soil properties, specially soil moisture and bulk density would be variables around the year
Author Response
Answers to Reviewer #1
General answers
Manufacturers of EM recommend using them to improve the properties of soils: (i) biological, (ii) chemical, and (iii) physical. The most scientific papers describe changes in the biological and chemical properties of soils - model potting and short-term ones. There are few articles describing changes in physical properties.
Various soil types (Reference Soil Groups) were tested in the world. Our studies supplement information about Haplic Luvisols RSG. These soils are common in the world, therefore the issue described is not marginal. Lack of data on soil physical condition prompted us to undertake research in this area. Our study verifies the hypothesis that soil inoculation with EM (EM-A) improves physical properties. Long term study (5 years) eliminates incorrect reasoning.
Samples were taken using methods generally accepted in soil science (Field Book for Describing and Sampling Soils, by National Soil Survey Centre Natural Resources Conservation Service U.S. Department of Agriculture). Scheme below.
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Type of samples |
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Soil with natural structure – metal cylinders |
Soils wit natural structure in boxes |
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Air-water properties:
BD, TP, FAC, FAP, FC, AWC, UWC, FC/TP |
Air dried, sieved |
Air dried |
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TOC, texture, PD |
Water-stable aggregates, MWD |
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We have supplemented the manuscript with tables.
We would like to keep the correlation results in the Results and discussion section.
Detailed answers
Line 12: word „biopreparatu” is changed on ‘biopreparat”
Line 23: Sentence: “Therefore, the soil application of such biopreparations as EM-A requires serious consideration.” is deleted from the Abstract.
We have added two new sentences:
- “It was an adverse effect.”
- “However, changes in soil compaction and air-water properties did not show significant deterioration.”
Lines 36-37: We changed the structure of the previous sentence to: “They are the degradation and mineralization of soil organic matter that are regarded as the most important among these processes.”
Lines 42-43: We have written the sentence more precisely.
Lines 66-69: The authors of the cited research did not add any substrates to the soil.
Line 97: Yes. As the TOC content increases, soil water retention increases, then hydraulic conductivity decrease.
Lines 151-155: According to Reviewer’s suggestion we have moved the paragraph to the beginning.
Lines 137-150: According to Reviewer’s suggestion, we have changed order of description of the crop rotation and EM application.
Line 163: We agree that the physical properties are very variable and depend on soil conditions.
That is why samples we collected samples in 10 replications on 3 dates. See the section “Materials and methods”.
Reviewer 2 Report
The paper entitled Impact of Effective Microorganisms (EM) application on the physical condition of Haplic Luvisol by Pranagal et al. looks at the effects of a three years of EM application on the physical condition of Haplic Luvisol. The authors measured a wide range of soil properties, including particle density, total organic carbon content, bulk density, total porosity, air capacity, air permeability, soil moisture at sampling, field water capacity, available water content, unavailable water content, water-stable aggregates content and FC/TP ratio. TOC and water-stable aggregates appeared to be the most sensitive to this treatment in comparison to other soil physical properties.
Abstract. Overall requires re-writing and structuring. While the hypothesis and the results were vaguely mentioned, I highly recommend re-writing it the abstract to flow easier. It seems disconnected with grammatic mistakes. The authors also didn’t include the implications f such study. Is the less of TOC and water aggregates a beneficial thing or not? Also adding a sentence about the rationale of the study is a good addition.
Line 11 add of before a 5 year application
Line 11-12 reconsider structuring this sentence “….the content of 11 water-stable aggregates and organic carbon as an effect a 5-year application of microorganisms”
Line 12, .replace by .that long-term application of EM-A biopreparat has
Line 14-14: restructure:” . The soil samples, their 15 natural structure preserved intact, were collected three times each year.”
Line 18 spell out FC/TP
Introduction: requires significant English editing. The rationale of the study was not highly justified. The authors mention that the full rationale of their study is that the authors of multiple studies suggested that “the full assessment of the effect of EM biopreparations on the soil environment requires multi-year research in various soil-climate conditions”. I don’t think the paper is addressing the various soil-climate conditions as it is not comparing between multiple sites. As such the research justification appears to be very weak.
Line 29, what do you mean by soil environment data?
Line 29-30: re-structure the first full sentence, for example : Environmental soil data indicate that arable lands are increasingly exhibiting various forms of soil degradation, thus implementation of special measures is needed.
Line 31,flow of the sentence is not clear, also add a comma after remediation
Line 36 , replace they are by it is
Lone 38 transformation of soil organic matter rather than transformations
Line 41-42 please re-structure
Please restructure the following lines
Line 62 , Lines 78-79, line 115
Line 68 on the level?
Materials and methods:
Iine 130: restructure the last part of the sentence.
I don’t think figure 1 serves the purpose. I think adding image or Arial photography form the field makes more sense,
Line 138 on April 11
Results and discussion:
The results and discussion section is too long. The author should consolidate it. Authors spent so much time explaining trends even though they are not significant. The authors explained each parameter separately in its own section making it too lengthy without properly linking all the measurements together, The authors could represent the same data in a much shorter paper with no need to spend 1-2 paragraphs talking about trends that are not significant. Just focus on the significant trends and explain what they mean. Also having the same color scheme will be more helpful. How about using some PCA or NMDS plots as well. It will help you bring the study together rather than having to plot each parameter alone.
Line 159, how do you the natural structure was preserved. Or what do you mean by that?
Figure 3, if you had multiple replicated per year, can you add error bars to these graphs
Also the colors are very hard to distinguish
It will be hard to justify it is accelerated mineralization of the organic matter without further data. Did you measure fluxes for example? How about runoff? Etc
The authors keep mentioning Table 5 but there is no table 5 included. I actually don’t see any tables. The authors mention so many tables but not in the right order and the tables can’t be found
Line 269-271 but it is not significant. Also please add error bars
Lines 580-587: I am still not sure how to interpret these results. Are they good, bad, beneficial, etc? I think it will be worth to compare these results to yield. If you are going to test EM-A and its effects on soils, it will be important to see how it correlates with yield as well.
Ok lines ? 586 include what I was asking about which should be include din the abstract as well. To say that EM is useful or not, requires more data and not just physical properties. It will have to be better linked with yield, economical benefits etc. Also, it will be worth to look at the microbial community composition in the soils before and after addition. The original composition of SOM and microbial communities in the soil will play a big role in how beneficial EM will be and thus can’t be generalized to all fields etc. So I recommend that the authors clarify some of these biases
In summary, while I have no problem with the data provided in this paper, the presentation is weak and requires restructuring of the whole paper and a better incorporation of the data.
Author Response
Answers to Reviewer #2
General and detailed answers (point by point)
According to Reviewer’s suggestion we have edited the abstract and completed it with sentences.
“It was an adverse effect.”
“However, changes in soil compaction and air-water properties did not show significant deterioration.”
“This research addresses the data gaps in the EM application.”
Lines 11-12: We have edited the sentence.
Lines 14-15: We have edited the sentence.
Line 18: Done.
Introduction
Language: The text was translated by a professional translation agency. We have corrected typing errors. They probably have resulted from the auto-correction of the text editor.
We understand that our research concerned only one type of soil (one RSG) and was conducted in one place. The studies supplement the information on Haplic Luvisols in a temperate/transitional zone, with a strong influence of the continental climate.
Line 29: Soil science assumes that the soil environment is a system of three phases: solid, liquid and gas. The soil environment is treated as a synonym for soils.
Lines 29-30: Done.
Line 31: We have edited the sentence.
Line 36: We have edited the sentence.
Lines 41-42: We have edited the sentence.
Line 62: Changed.
Line 68: We have changed „level” to „quantity”.
Lines 78-79: We have edited the sentence.
Line 115: Corrected.
Line 130: We have edited the sentence.
Figure 1. Changed. We would like it to stay at work. It presents well the scheme of field experiment.
Line 138: We have changed „in” to „on”
Results and discussion
In our opinion, a thorough discussion of the results well presents all trends, even those more or less significant.
After discussions in our team, we would like to keep the established colour scheme and chart type.
Line 159: We have edited the sentence. It is more clear, now.
Figure 3. We have changed colours. We gave up adding error bars on the charts, because we supplemented the work with result tables (tables 1-3).
Changes in carbon content are given in the Table 1. We know from many years of experience that soil organic matter is very stable and the proportion of water-soluble humus fraction is small. Runoff was not included because the experimental field was completely flat.
The work was supplemented with tables 1-5.
Lines 269-271: At work, we wanted to present both significant and insignificant differences in results. This is a complete set and shows how soil properties change.
Lines 269-271: We gave up adding error bars in charts because tables 1-5 were added.
Lines 580-587: Due to the volume of work, we gave up presenting the results of plant crops, soil organic matter composition, number and share of microorganism colonies. We have written about this in a comprehensive introduction, citing the research of other authors. We decided to carry out research mainly on the physical properties of soils that change under the influence of EM applications, since this type of work was scarcely revealed. Especially works describing many years of research conducted as field experiments.
Round 2
Reviewer 1 Report
The paper “Impacts of effective microorganisms (EM) application on the physical conditions of Haplic Luvisol”, analyses the impact of effective microorganisms’ application to soil physical properties in a field essay along five years. In this work the EM application is annually repeated in a crop rotation essay along 5 years. In each one of the analysed years the soil management was different as dictated by crop rotation and for this reason the year is considered as an additional variability factor.
This is the second review, the first one was incomplete (the results could not be reviewed) because the authors did not include the tables in the paper.
The study is a hard work and has intensive data collection for several years but still some corrections must be made before the manuscript must be published. Statistical analysed must be defined and centred about the effects of EM application on soil physical properties.
Specific comments.
Lines 12-13. The hypothesis that long-term applied EM-A 12 biopreparatu have a positive effect on the soil physical condition has been verified.
It is not correct since the results show that physical properties are worse in treated plots
Lines 42 -43. Change “They are the degradation and mineralization of soil organic matter that are regarded…” by : “The degradation and mineralization of soil organic matter is regarded…”
Line 44 EM-A is not previously defined
Line 70 Change “An effect” by “The effect”
Line 101. What is Greek manure?
Lines 124-125. Same indications as lines 12-13
Lines 183-184. It is not clear how many samples were taken in each sampling date and each plot.
Lines 237-239. Which variable is used as a factor in one way ANOVA? The results of two way ANOVA are not presented. Which statistical program was used to analyse the data?
Line 244. If the total number of samples during the experience is 150, as you calculate CV separately for treated and control plot n = 75
For the results presented in tables 1 to 3 each number is not a single value, is a mean value. Please, indicate in each table the number of samples used to obtain each value and also the variability
Line 261 Is it statistically significant?. In general, you did not show statistical significance
Figures 3 to 6. Please indicate the number of samples. In the figures, only the interaction is compared but not individual comparison of factors is presented. The factor year is mixed with the factor EM application.
Lines 279- 291. In general, the presentation of the results is not linear. Argumentation moves from one table to another in the same paragraph. I suggest moving this paragraph and similar paragraphs regarding other properties to a new section where the correlation between the variables is explained.
Lines 626-628. The differences may be due to in the first essay EM were applied jointly with organic matter
Lines 645- 646. You can only say that the repeated applications of EM are questionable because they accelerate the decomposition of soil organic matter
Author Response
Responses to the Reviewer #1
Lines 12-13:
According to suggestion of Reviewer we have changed the final part of the sentence from “[…] has been verified” to “has not been confirmed”.
Lines 42-43 (43-44 now):
The sentence has been changed from – “They are the degradation and mineralization of soil organic matter that are regarded […], to “The degradation and mineralization of soil organic matter are regarded […].
Line 44 (45-46 now):
According to suggestion of Reviewer we gave the full name of “effective microorganisms” and the next the abbreviation – (EM-A).
Line 70 (71 now):
We have changed from “An effect” to “The effect”.
Line 101 (102 now):
It should be “green manure”. The MS Word has done auto-correction without our knowledge.
Lines 124-125 (125-126 now):
According to Reviewer suggestion we have changed the sentence
“The hypothesis that long-term applied EM-A biopreparat have a positive effect on the soil physical condition has been verified.”
to – “We put the hypothesis that long-term applied EM-A biopreparat have a positive effect on the soil physical condition”.
Lines 183-184 (186-188 now):
We have provided how many samples were collected from each plot in each sampling date.
We have changed the sentence:
“Soils samples were extracted from the 0-10 cm layer, in ten replicates, to 100 cm3 metal cylinders.”
To: “Soils samples were extracted from the 0-10 cm layer to 100 cm3 metal cylinders, in two replicates from five TEM and five Control plots (Fig. 1), in each sampling date (n = 20). 300 soil samples were collect during 5 years.”
Lines 237-239 (243-245 now):
The Control and TEM were used as the factors in one-way ANOVA. We have supplemented this information in the lines 243-245. The results of one-way are presented in the table 4.
The results of the two-way ANOVA, Tukey’s and the lowest significant difference (LSD) tests we have presented in the Figures 3-8, as combination of letters a, b, c, ab, ac, bc, etc., above chart bars. The same letters indicate no difference between analysed properties.
Statistica 11 by Statsoft and ARSTAT by University of Life Sciences in Lublin were used to statistical analyses. We have added this sentence to 2.3. Statistical analysis section, lines 254-255.
Line 244:
According to the comments of the reviewer, we have calculated again the number of samples in the experiment. The total number of samples was 300, variation coefficients (CVs) were calculated separately for TEM (n = 150) and Control (n = 150). This information has been put in the right lines.
Number of replicates: Control – n = 10, TEM – n = 10, i, ii, iii – n = 20, I,…V – n = 60. This information we have introduced in the lines: 187-188, 250-251, 267, 280, 597.
Line 261:
The differences are shown in Fig. 3 and in tab. They are letters above the chart bars and at values in the table. They indicate significant differences. Description of how to interpret significance is given in the caption under the figure and table (line 281, lines 601-603). The system is commonly used to present significant differences between means.
The number of samples is added under the figure (line 280).
Reviewer: “The factor year is mixed with the factor EM.”
Answer: The factor year (1) is mixed with factor treatment (2): a. EM application or b. without EM application.
The differences between sampling dates (i, ii, iii) can be analyzed on the base of data in Tables 1-3.
Lines 279-291:
We would like to leave the current data description scheme. We believe that this form of description is a complete discussion of the results and discussion for each soil property.
Lines 626-628:
We agree with the reviewer's opinion that the TOC supplement could have influenced the results in that experiment (of other authors). We believe that laboratory results should not be generalizations. Model studies in soil science should be used to prepare a field experiment, for example, determining the dose of fertilizers or other additives (compost, sewage sludge, biochar, waste, etc.).
Lines 645-646:
As suggested by the reviewer, the previous sentence has been completed and is now as follows:
Therefore, the application of such biopreparations as EM-A requires serious consideration, because they accelerate the decomposition of soil organic matter. Carbon sequestration in soil humus is positive phenomenon. It increase resistance to physical degradation.
General comments
After analyzing the reviewer's comments, we sorted out the titles of the tables and explanations below them.
Reviewer 2 Report
I would like to thank the reviewers for some of the adjustments. I still feel this is a long paper with lots of un-needed discussion but I also respect the authors decision to keep all of it to provide a deeper look into soil properties. There are some areas that still require some English proofing especially in the abstract where some of the newly added sentences feel disconnected.
Author Response
Responses to Reviewer #2
Thank you for all the comments that have raised the substantive quality of our work. We appreciate your understanding of the importance of the current form and length of the text.
We would like to kindly inform you that we have corrected the English language and corrected the abstract text.
