The Characteristics and Influential Factors of Earthworm and Vermicompost under Different Land Use in a Temperate Area, China

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Ms. Titled The Characteristics and Influential Factors of Earthworm and Vermicompost underDifferent Land Use in a Temperate Area, China

 Li Ma, Ming’an Shao, Yunqiang Wang, Tongchuan Li, Xuanxuan Jing, Kunyu Jia and Yangyang Zhang

The ms presents the results of research on earthworms and the properties of vermicompost in 6 ecosystems differing in the type of land use. The authors point out that there are no comprehensive studies on how the soil nutrient composition and bacterial communities change in response to earthworm community natural invasion in artificial forests and cropland. Further they presented that the main aim of the research was to: (1) investigate earthworm communities (species and populations) in temperate natural ecosystems in China, which it will provide basic data for earthworm research; (2) examine the main factors influencing the earthworm communities and vermicompost distribution; (3) exploring the C, N accumulation in soil from the perspective bacterial roles, in order to reveal the role of earthworm in soil nutrition accumulation and mineralization in different land use types.

The presented study although interesting in the context of interaction among earthworms, vermicompost and microbial communites requires clarification and improvement.

 There is no clearly stated research hypothesis

 The text contains inconsistencies, e. g. why in the abstract the authors indicate that they studied artificial ecosystems, and then indicated that the main research goal was to assess the species composition in temperate natural ecosystems in China.

 Line 18 what do the authors understand by the term natural invasion of earthworms, since later in Line 21 they mention what species of earthworms dominated in the communities. This indicates that community composition was analyzed per study site.

 Line 107 – 110 it is unclear how the earthworms were collected, whether by manual sorting or by using an electric shock?

 Why was the significance of the differences in earthworm population density and vermicompost weight not statistically tested? Were the RDA models statistically significant?

 

Line 245-247  – unclear which species of earthworms were classified into ecological categories, i.e. which were classified as epigeic, anecic, etc. Epi-endogeic earthworms (please add the ecological classification of earthworm species to Supplementary materials).

 Line 246 – Epi-endogeic  which one?

 Line 245 – 250 - there is a lack of continuity in the argument

Line 251 – population – do you mean population density?

Line 262 – do you mean difference in earthworm community composition?

Table 2 – indication sp. nov/sp. Nov according to Zoological Nomenclature – species without diagnosis is nomen nudum!

Table 2 - please show the abundance of each species ± of what standard error or standard deviation ? in the Table. The same explanation is need in the Table 3.

 

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Authors have reported how the soil nutrient composition and bacterial communities change in response to earthworm community natural invasion in artificial forests and cropland. The paper is simple, straightforward, and well-written.

Comments

1.      Abstract: “Earthworm communities influence soil carbon and nitrogen circulation by altering the diversity and composition of microbial communities, which is improving soil fertile.” - It should be “which improves soil fertility.”

2.      Abstract: “Earthworm community structure plays key roles in decreasing of bacteria diversity and add special bacteria in vermicompost……” Instead of writing "special bacteria," please mention the dominant bacteria.

3.      Table 1: Edit the typo “/” before %. Why is there only one value, with no replicates? No standard error, or significance test? Please provide at least 3 replicates from three points in the field to obtain unbiased data.

4.      Table 2: It might be better to add diversity for each species individually and then together to get clearer data and check for significance.

5.      2023 y: If “y” means “year,” please clarify it in the table footnote.

6.      Fig. 1 Legends: Please provide the full form of all abbreviations used in the figure for ease of reading.

7.      Table 3: I am curious to know how the authors identified diversity. Since they did not isolate and sequence the species, they only did metagenomics of the soil.

 

8.      There are several inconsistencies in font, spacing, and grammatical errors. Please check carefully.

Comments on the Quality of English Language

Minor editing of the English language is required as there are some inconsistencies in font, spacing, and grammatical errors.

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

I think the main problem in this MS in the experimental design and its performance

On which base did the author select these land-use types? Which link can be found among these types?

Why the authors use abbreviations are not link to the names of these land use-types:

“Six representative land-use types named Indus Forest (WS), walnut forest (TS), apple

orchard (GS), kiwi orchard (MS), ryegrass land (CS), and corn fields (YS) in the Baoji region were selected for field surveys.”

Basically, before starting this study, more than soil texture or water content (Table 1), should be mentioned especially the nutrient status of these soil and microbial parameters

What about the historical background of each soil type (land use type)?

When this study starts and when finish?

In Figure 2, there is no meaning of all abbreviations

“Figure 2. The characteristics of bacterial classification in soil(a) and vermicompost (b) under different land use types.  

 

For the previous reasons and the MS is not mature enough to publish.

Thanks

Author Response

I think the main problem in this MS in the experimental design and its performance

On which base did the author select these land-use types? Which link can be found among these types?

Response: Thank you for your comments.

These land-use types are the typical land in the Baoji region. We found that earthworm’s active and numerous populations live in these vegetation ecosystems by large number of field surveys.

Why the authors use abbreviations are not link to the names of these land use-types: “Six representative land-use types named Indus Forest (WS), walnut forest (TS), apple

orchard (GS), kiwi orchard (MS), ryegrass land (CS), and corn fields (YS) in the Baoji region were selected for field surveys.”

Response: thanks for your suggestion.

We named the land use names by using the initials of land English names. Six representative land-use types named buttonwood forest (BS), walnut forest (WS), apple orchard (AS), kiwi orchard (KS), ryegrass land (RS), and corn fields (CS).

Basically, before starting this study, more than soil texture or water content (Table 1), should be mentioned especially the nutrient status of these soil and microbial parameters

What about the historical background of each soil type (land use type)?

Respond：Thank you for your comments.

the nutrient status of these soil and microbial parameters of each soil type were showed in figure 1 and figure 2.

When this study starts and when finish?

Respond：Thank you for your comments.

The study started on June 2022 and finished April 2023.

In Figure 2, there is no meaning of all abbreviations

“Figure 2. The characteristics of bacterial classification in soil(a) and vermicompost (b) under different land use types.  

Respond：Thank you for your comments.

Figure 2. The characteristics of bacterial classification in soil(a) and vermicompost (b) under different land use types.（Soil: buttonwood forest（BS1,BS2 ,BS3）,walnut forest（WS1,WS2,WS3）, apple orchard（AS1,AS2,AS3）, kiwi orchard（KS1,KS2,KS3）, ryegrass land（RS1,RS2,RS3）, corn field（CS1,CS2,CS3）; Vermicompost: buttonwood forest（BV1,BV2,BV3）,walnut forest（WV1,WV2,WV3）, apple orchard（AV1,AV2,AV3）, kiwi orchard（KV1,KV2,KV3）, ryegrass land（RV1, RV2, RV3）, corn field（CV1,CV2,CV3））

Reviewer 4 Report

Comments and Suggestions for Authors

General comments

·                  In overall, I found the work very interesting and in line with the Journal scope. Presentation and writing style is good with some lapses in style. Structure is clear.

·                  Title: appropriate

·                  Abstract: it well written and main data results were included.

·                  Methodology: Good description, coherent and detailed. Apart from the fact that it is not clear the experimental set-up in the field.

·                  References: >63% οf References are up-to-date.

 

Specific comments (list not exhaustive)

Introduction:

Are there any studies of vermicompost of such earthworms species with biowastes? If so they could be mentioned

Materials and Methods:

It was not clear. The experimental set-up was in the landscape?

Results:

L149: correct Eisenia fetida and all through the manuscript

L158-159: Rephrase

L165, L181-182, L193: correct: (Fig. 1 (a1, a2)) consistent depiction of Figures

L171-175: please add data for comparison reasons

Fig1: what are the letters “a, b, c, d” on the graphs depict? Clarify

L199-200: rephrase

Fig.3: please use the same numbers of x- and y-axis (for comparison reasons)

Discussion:

L254, L264, L266, L276, L290: rephrase/ correct

L269, L298, L299: correct Table 2

In Tables regarding units correct”/” with “(..)”

Table 4: Data (correlation analysis) should be presented in a better way

On the basis of the above, my recommendation is a Minor revision and I hope the authors define my perspective useful in improving their manuscript.

 

Comments on the Quality of English Language

Author Response

In overall, I found the work very interesting and in line with the Journal scope. Presentation and writing style is good with some lapses in style. Structure is clear.

• Title: appropriate
• Abstract: it well written and main data results were included.
• Methodology: Good description, coherent and detailed. Apart from the fact that it is not clear the experimental set-up in the field.
• References: >63% οf References are up-to-date.

 

Specific comments (list not exhaustive)

Introduction:

Are there any studies of vermicompost of such earthworm’s species with biowastes? If so, they could be mentioned

Respond：Thank you for your comments.

We add the studies in vermicompost technology as biowastes. “It has been reported that vermicomposting technology wildly used for agricultural and industrial wastes due to efficient nutrient recovery potential and pollutant remedial role.”

[24] Devi J., Pegu R., Mondal H., et al. Earthworm stocking density regulates microbial community structure and fatty acid profiles during vermicomposting of lignocellulosic waste: unraveling the microbe-metal and mineralization-humification interactions. Bioresource Technology, 2023,367. doi:10.1016/j.biortech.2022. 128305.

 

Materials and Methods:

It was not clear. The experimental set-up was in the landscape?

Respond：Thank you for your comments.

Six representative land-use types named buttonwood forest (BS), walnut forest (WS), apple orchard (AS), kiwi orchard (KS), ryegrass land (RS), and corn fields (CS) in the Baoji region were selected for field surveys. In each land-use type, we established five randomly distributed 1.0 × 1.0 m plots for determining earthworm community and vermicompost characteristics. Earthworms were collected by excavating and manual sorting in soil. All earthworms that appeared on the surface were collected, and the soil adhering to the body surfaces of worms was washed away. Sexually mature earthworms of different colors were selected and preserved in ethanol for gene detection with tagging to determine the species. In each plot, soil surface vermicompost was collected manually, and soil samples were collected at 10-cm-depth intervals from 0 to 100 cm using a soil auger. Surface impurities were removed from soil samples by passing through a 0.25 mm or 2 mm sieve and the sieved samples were thereafter used to determine the pH and the contents of soil organic carbon (SOC), and total N. SOC was determined using the potassium dichromate oxidation-external heating method; total N was determined using the Kjeldahl method. Meanwhile, Soil water content, soil particle-size distribution, pH and bulk density were measured in different use land soil. The average values of physical and chemical properties of soil samples collected from these sites are listed in Table 1.

Results:

L149: correct Eisenia fetida and all through the manuscript

Respond：Thank you for your comments.

Eisenia fetida changed to “Eisenia foetida”

L158-159: Rephrase

Respond：Thank you for your comments.

Change to “It is a significant positive correlation between the weight of the vermicompost and earthworm density in the same season under different land uses. There are more earthworm populations in artificial forests, such as, AS, WS and CS in spring.”

L165, L181-182, L193: correct: (Fig. 1 (a1, a2)) consistent depiction of Figures

Respond：Thank you for your comments.

They were modified to “Fig. 1 (a1, a2), Fig. 2 (b₁, b₂) , Fig. 3 (c₁, c₂)” in the paper.

L171-175: please add data for comparison reasons

Fig1: what are the letters “a, b, c, d” on the graphs depict? Clarify

Respond：Thank you for your comments.

Different lower-case letters indicate significant differences in treatment at the 0.05 level.

L199-200: rephrase

Respond：Thank you for your comments.

“The diversity of soil bacterial communities with increases in earthworm populations (table 2 and table 3).” Change to” It is benefit for abundant diversity of soil bacteria by increasing earthworm populations (Table 2 and Table 3).”

Fig.3: please use the same numbers of x- and y-axis (for comparison reasons)

Respond：Thank you for your comments.

b

a

Figure 3. RDA (Redundancy Analysis) map of relationship between earthworm population density, environmental factors and bacterial community in earthworm compost. （a:soil; b:vermicompost）

 

Discussion:

L254, L264, L266, L276, L290: rephrase/ correct

Response: Thanks for your suggestion.

L254: “The moderate temperature of earthworm activity was from 10 ℃ to 40 ℃” change to “The optimal temperature for earthworms was 10 ℃ to 40 ℃.”

L264: “lower temperature of last winter limited earthworms’ excretion activities on the surface resulting in less vermicompost in following spring.” change to “In spring, there was less vermicompost accumulation on the surface because of limitation earthworms’ excretion activities by the lower temperature of last winter.

L266: “The number of vermicompost could geratly affect soil structure, and nutrient distribution in directly” change to “The number of vermicompost could impact on soil structure and nutrient distribution, directly.”

L276: “Such as, their vermicomposting TS (766 g/m ²) and in MS (732.7 276g/m ²) in summer, in WS (240.8 g/m ²) and in MS (235.4 g/m ²) in spring (table 1)” change to “Such as, the vermicomposting of TS (766 g/m ²) and MS (732.7 276g/m ²) in summer, and WS (240.8 g/m ²) , MS (235.4 g/m ²) in spring (Table 1).

L290:“Comparing the surrounding soil, total N content of vermicompost has been improve by 1.6 to 7.7 times, respectively” change to“Comparing the surrounding soil, the total N content in vermicompost enhanced by 1.6 to 7.7 times in different land use, respectively.”

L269, L298, L299: correct Table 2

In Tables regarding units correct”/” with “(..)”

Response: Thanks for your suggestion.

L269, L298, L299: “table 2” changed to “Table 2”.

We have changed the units in Table 2

Table 4: Data (correlation analysis) should be presented in a better way

Response: Thanks for your suggestion.

We have changed the units in Table 4

 

Author Response File: Author Response.pdf

Reviewer 5 Report

Comments and Suggestions for Authors

Dear authors!

The article is devoted to the current problem of soil fertility in areas occupied by various artificial forests and croplands, under the influence of the natural earthworm community. The subject of the manuscript is relevant both for the region of study and for other regions. The article contains an analysis of the proportionate contribution of vermicompost and earthworms to soil nutrition accumulation and mineralization in different land use types in the natural eco-environment. However, there are a number of important comments. I recommend the authors correct the article. The article can be accepted after a minor revision.

1.      In the Abstract section, the purpose of the study should be formulated more clearly.

2.      Latin names should be in italics. Please look throughout the text.

3.      To designate the same type of forest, the authors use different terms in the article : “buttonwood forest” (P.1 l.20; P.3 in Table 1.) and “indus forest” (P.3 l.96; P.7 l.202, 203). I recommend choosing one.

4.      In the Introduction section (P.2 l.63), the authors refer to the book by Lavelle & Spain (2001) and indicate “referred to “vermicompost,” the output of which has been estimated from 36 to 108 mg ha-1 yr-1 in temperate areas". It is obvious that there is an error in the units of measurement here.

5.      (P.3 l.103-104) The authors indicate that “all earthworms that appeared on the surface were collected.” It should be indicated for what purpose this was done, since only certain varieties of earthworms tend to crawl to the surface, while other species tend to remain in the soil even when it is hydrated.

6.      “Experimental Design and Sample Preparation” does not provide methods for determining indicators that are discussed later in the text (P.3 l.100; P.9-10 l.231-246): “Soil water content”, “Particle -size distribution (Sand, Silt, Clay)", "Bulk density", "total porosity (PS)".

7.      It should be specified why soil samples were passed through a 0.25 mm or 2 mm sieve. What impurities were separated on each of the sieves?

8.      (P.10, l. 270) “Epi-endogecic” is probably a typo. It should be "epi-endogeic".

9.      The Conclusions section does not reflect the proportionate contribution of vermicompost to background-soil nutrients, whereas this is mentioned in the purpose of the study.

Author Response

The article is devoted to the current problem of soil fertility in areas occupied by various artificial forests and croplands, under the influence of the natural earthworm community. The subject of the manuscript is relevant both for the region of study and for other regions. The article contains an analysis of the proportionate contribution of vermicompost and earthworms to soil nutrition accumulation and mineralization in different land use types in the natural eco-environment. However, there are a number of important comments. I recommend the authors correct the article. The article can be accepted after a minor revision.

1. In the Abstract section, the purpose of the study should be formulated more clearly.

Response: Thanks for your suggestion.

We modified the Abstract section as follow:

Earthworm communities influence soil carbon and nitrogen circulation by altering the diversity and composition of microbial communities, which improves soil fertility. Studying the soil nutrient composition and bacterial communities change in response to earthworm community natural invasion may be key to exploring earthworm ecological functions and accurate assessing C, N mineralization in artificial forests and cropland. In this study, we examined the communities of five earthworm species in ecosystems characterized by six different land-use types, such as, buttonwood forest, walnut forest, apple orchard, kiwi orchard, ryegrass land, and corn field. The Metaphire baojiensis (new subspecies unpublished) and Amynthas carnosus planus (new subspecies unpublished) were dominant earthworm species. Among different land-use types, earthworm densities ranged from 2 to 27 ind·m^-2 in summer and 15 to 40 ind·m^-2 in spring. But surface vermicompost weight in summer (296.7 to 766.0 g·m^-2) was more than spring. There was a positive correlation between the weight of the vermicompost and earthworm numbers in the same season. The soil carbon (C), total nitrogen (N) of vermicompost ranged from 5.12 to 20.93 g·kg^-1, and 0.52 to 1.35 g·kg^-1, respectively. Compared with soil, the contents of vermicompost C, N increased 2.0 to 4.3 times, 1.6 to 7.7 times, respectively. The average C/N of vermicompost (9.5~23.5) was higher than them in soil (7.3~19.8). Due to the higher abundances of C, N in soil of corn fields and kiwi orchards, which are cultivating higher abundances of earthworms and more vermicompost. the C, N and C/N of vermicompost is higher than soil. The C and N were accumulated by earthworms’ excreting and feeding activity instead of vegetation in vermicompost. Earthworm community structure plays key roles in decreasing bacteria diversity and adding Proteobacteria, Actinobacteria, Acidobacteria, Bacteroidetes, Chloroflex in vermicompost result to enrichment soil C, N and increasing C/Nin vermicompost. So, the evaluation different vegetation ecosystems in soil C, N pools accumulation and mineralization should be paid more attention on the function of earthworm community in the future.

 

2. Latin names should be in italics. Please look throughout the text.

Response: Thanks for your suggestion. We have checked the text and modified them.

3. To designate the same type of forest, the authors use different terms in the article : “buttonwood forest” (P.1 l.20; P.3 in Table 1.) and “indus forest” (P.3 l.96; P.7 l.202, 203). I recommend choosing one.

Response: Thanks for your suggestion. We have kept the name of the same type of forest consistent and changed the abbreviation of land named in text. Six representative land-use types named buttonwood forest (BS), walnut forest (WS), apple orchard (AS), kiwi orchard (KS), ryegrass land (RS), and corn fields (CS).

4. In the Introduction section (P.2 l.63), the authors refer to the book by Lavelle & Spain (2001) and indicate “referred to “vermicompost,” the output of which has been estimated from 36 to 10⁸ mg ha^-1 yr^-1 in temperate areas". It is obvious that there is an error in the units of measurement here.

Response: Thanks for your suggestion. “36 to 10⁸ mg ha^-1 yr^-1” change to “36 to 10⁸ g·ha^-1·yr^-1”

5. (P.3 l.103-104) The authors indicate that “all earthworms that appeared on the surface were collected.” It should be indicated for what purpose this was done, since only certain varieties of earthworms tend to crawl to the surface, while other species tend to remain in the soil even when it is hydrated.

Response: Thanks for your suggestion.

We collected all earthworms by manual excavating in the soil depth from 0cm to 50cm.

So, this sentence change to “Earthworms were collected by excavating and manual sorting in soil depth from 0cm to 50cm and the soil adhering to the body surfaces of earthworm was washed away.”

6. “Experimental Design and Sample Preparation” does not provide methods for determining indicators that are discussed later in the text (P.3 l.100; P.9-10 l.231-246): “Soil water content”, “Particle -size distribution (Sand, Silt, Clay)", "Bulk density", "total porosity (PS)".

Response: Thanks for your suggestion.

It was supplemented as “Six representative land-use types named buttonwood forest (BS), walnut forest (WS), apple orchard (AS), kiwi orchard (KS), ryegrass land (RS), and corn fields (CS) in the Baoji region were selected for field surveys. Five sample points were randomly selected from different use land and the soil was collected at 0-30cm depth. Soil water content, soil particle-size distribution, pH and bulk density were measured in different use land soil. The average values of physical and chemical properties of soil samples collected from these sites are listed in Table 1.”

7. It should be specified why soil samples were passed through a 0.25 mm or 2 mm sieve. What impurities were separated on each of the sieves?

Response: Thanks for your suggestion.

The aim was to screen out unnecessary components such as coarse particles and impurities from the soil, retaining fine soil particles and nutrients, in order to improve the accuracy of measurement.

“…soil samples were passed through a 0.25 mm or 2 mm sieve” change to “…soil samples were passed through a 0.25 mm sieve”

8. (P.10, l. 270) “Epi-endogecic” is probably a typo. It should be "epi-endogeic".

Response: Thanks for your suggestion.

“Epi-endogecic” change to “epi-endogeic”

9. The Conclusions section does not reflect the proportionate contribution of vermicompost to background-soil nutrients, whereas this is mentioned in the purpose of the study.

Response: Thanks for your suggestion.

The Conclusions section was modified as follow:

In this study, we established that different patterns of land use can influence the structure of earthworm communities and the distribution of soil nutrients dependent on earthworm behavior (vermicompost). Earthworm community structure was in turn found to influence the composition of soil nutrients, with positive correlations being observed between earthworm density and the contents of C, N, and soil water. In addition, the distribution and accumulation of C, N were found to be influenced by earthworm community activities. Soil organic C, and total N were enriched in the vermicompost through earthworm community activity, and in turn, vermicompost contributed to altering the bacterial community diversity. The carbon content of vermicompost is 2.0 to 4.3 times higher than that of soil. Comparing total nitrogen content of soil, total nitrogen content increases 1.6 to 3.7 times in vermicompost. Furthermore, we established that the diversity of bacteria in vermicompost was lower than that in soil, which can have the effect of retarding the conversion rates of C and N, thereby resulting in long-term nutrient accumulation in the soil. In addition, vermicompost was found to be populated by a number of beneficial bacterial species, including those in the phylum Cyanobacteria, Verrucomicrobia and Patescibacteria which play an important role in nutrient fixation, and thus contributing to accumulation these nutrients in the soil. Collectively, our findings confirm that in addition to the influence of environmental factors, earthworm community structure plays key roles in determining bacterial community composition and activity in vermicompost. There are important affection on the soil C, N pools accumulation and mineralization by earthworm communities in different natural ecosystems.

 

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Unfortunately , I cannot accept this work. The authors have no basic knowledge of the widely accepted convention in zoology that rules the formal scientific naming of organisms .

In Table 2 - they swapped species e.g.

Amynthas carnosus planus sp. nov,

Metaphire baojiensis sp. Nov.

To

Amynthas carnosus planus nom. nud,

Metaphire baojiensis nom. nud.

In what sense did the authors use sp. Novum - whether as a new occurrence or as a species new to science. The previous representation of the species composition was erroneous and the current improvement is an absolute mistake.

When quoting the occurrence of a species - we quote according to the

International Code of Zoological Nomenclature

the recognised name of the species and the author who described the species and the year in which he described it.

E.g. Lumbricus rubellus Hoffmeister, 1843

There is no reference in the text as to who compiled the taxonomically collected earthworm material, according to which keys it was labelled.

Other comments in pdf

Comments for author File: Comments.pdf

Author Response

Thank you for your comments. The genetic detection of earthworm species came from the Instrumental Analysis Center of Shanghai Jiao Tong University. We have invited an expert from this Center to check the name of earthworm in the report. He has carefully reviewed the earthworm named in manuscript and amend the name of earthworm species in table 2 by your suggestions.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

thanks for your improvements

Round 3

Reviewer 1 Report

Comments and Suggestions for Authors

The species names of Lumbricidae are still incorrectly cited in Table 2!

The rules for reporting species composition are still not applied. I ask the authors to familiarize themselves with the International Code of Zoological Nomenclature and the basic principles of binomial nomenclature according to  Linnaeus.

Author Response

Reviewer #1:

There is no clearly stated research hypothesis

 The text contains inconsistencies, e. g. why in the abstract the authors indicate that they studied artificial ecosystems, and then indicated that the main research goal was to assess the species composition in temperate natural ecosystems in China.

Answer: Thank you very much for your comment.

In this study, we investigate earthworm diversity in six different land use, such as, Buttonwood forest (WS), Apple orchard (GS), Walnut forest (TS), Ryegrass land (CS), Kiwi orchard (MS), and Corn field (YS). These six different land uses are artificial ecosystems. So, I change the” in temperate natural ecosystems in China” to “in temperate artificial ecosystems in China”

 Line 18 what do the authors understand by the term natural invasion of earthworms, since later in Line 21 they mention what species of earthworms dominated in the communities. This indicates that community composition was analyzed per study site.

Answer: Thank you very much for your comment.

“the term natural invasion of earthworms” means the self-reproduction and living of earthworms in natural conditions in different ecosystems.

 

 Line 107 – 110 it is unclear how the earthworms were collected, whether by manual sorting or by using an electric shock?

 Why was the significance of the differences in earthworm population density and vermicompost weight not statistically tested? Were the RDA models statistically significant?

 Answer: Thank you very much for your comment.

• Earthworms were collected by excavating soil sample and manual sorting. We had modified Materials and Methods in the paper.
• We supplemented the statistically tested of significance of the differences in earthworm population density and vermicompost weight in table2.

3)  The Levene's T-test and in the part 2.3. Data and Statistical Analysis

Sequential denoising and amplicon sequence clustering based on amplicon sequence variants (ASVs) were performed according to the QIIME 2 data analysis process. The Chao1 index, Shannon and Simpson indices were evaluated the diversity of soil microbial communities. Statistical analyses were performed using SPSS Statistics for Windows (version 26.0; IBM Corp., Armonk, NY). One-way analysis of variance (one-way ANOVA) was used to compare the density of the earthworm community, nutrients, and diversity of the microbial community (significance level: p<0.05). The independent sample Levene's T-test was conducted to determine the differences in earthworm population density and vermicompost weight (significance level: p<0.05) to assess the effects of different land use. The impacts of environmental factors and earthworm density on bacterial communities were analyzed using redundancy analysis (RDA, Canoco 5). Origin software (version 2022b) was used to create the figures.

 

Line 245-247  – unclear which species of earthworms were classified into ecological categories, i.e. which were classified as epigeic, anecic, etc. Epi-endogeic earthworms (please add the ecological classification of earthworm species to Supplementary materials).

Answer: Thank you very much for your comment.

 Line 246 – Epi-endogeic  which one?

Answer: Thank you very much for your comment.

Amynthas is classified as Epi-endogeic. Metaphire is classified as anecic. Esienia fetida is classified as epigeic. We add the 7^th reference in line 247 about the ecological classification of earthworm species. [7. Nicolas Bottinelli, An explicit definition of earthworm ecological categories – Marcel Bouché’s triangle revisited, Geoderma, 2020]

 

 Line 245 – 250 - there is a lack of continuity in the argument

Answer: Thank you very much for your comment.

The part changed as follow:  “Earthworms can be classified into three ecological categories, namely, epigeic, endogeic, and anecic earthworms (Bouché, 1977), that differ with respect to living habits and ecological service functions (Guhra et al., 2020). On the Guanzhong Plain, most of the earthworm communities examined comprised anecic and Epi-endogeic earthworms, such as species of Metaphire and Amynthas, whereas the epigeic earthworm Esienia fetida was found only in land planted with ryegrass and in kiwi orchards.”……

“It has previously been established that land usage influences the species and populations of earthworms and their behaviors on soil (Hedde et al., 2013). It is significant differences in earthworm community composition among the six assessed land-use types (p < 0.05) (table 2).”

 

Line 251 – population – do you mean population density?

Answer: Thank you very much for your comment.

We changed the “population”to “population density”

 

Line 262 – do you mean difference in earthworm community composition?

Answer: Thank you very much for your comment.

“It is significant differences in earthworm abundance and species among the six assessed land-use types.” change to “It is significant differences in earthworm community composition among the six assessed land-use types.”

Table 2 – indication sp. nov/sp. Nov according to Zoological Nomenclature – species without diagnosis is nomen nudum!

Answer: Thank you very much for your suggetion.

We change the “sp. nov/sp. Nov” to “”nom.nud” in table 2.

 

Table 2 - please show the abundance of each species ± of what standard error or standard deviation ? in the Table. The same explanation is need in the Table 3.

Answer: Thank you very much for your comment.

We add “values are the mean + standard error” in footnote of table2 and table 3.

 

 

 

Reviewer #2:

1. Abstract: “Earthworm communities influence soil carbon and nitrogen circulation by altering the diversity and composition of microbial communities, which is improving soil fertile.” - It should be “which improves soil fertility.”

Answer: Thank you very much for your comment. We have revised the expressions as “…which improves soil fertility” in abstract.

2. Abstract: “Earthworm community structure plays key roles in decreasing of bacteria diversity and add special bacteria in vermicompost……” Instead of writing "special bacteria," please mention the dominant bacteria.

Answer: Thank you for your thoughtful comment. “Earthworm community structure plays key roles in decreasing of bacteria diversity and add special bacteria in vermicompost……” have changed to “ Earthworm community structure plays key roles in decreasing of bacteria diversity and add Proteobacteria, Actinobacteria, Acidobacteria, Bacteroidetes, Chloroflex in vermicompost….”

3. Table 1: Edit the typo “/” before %. Why is there only one value, with no replicates? No standard error, or significance test? Please provide at least 3 replicates from three points in the field to obtain unbiased data.

Answer: Thank you very much for your comment. We add the standard error in table 1, as follow:

Table 1. Soil physical and chemical properties at the experimental site.

4. Table 2: It might be better to add diversity for each species individually and then together to get clearer data and check for significance.

Answer: Thank you very much for your comment. Due to the limitation of field experiment conditions, we did not do numbers of each species statistical work.

5. 2023 y: If “y” means “year,” please clarify it in the table footnote.

Answer: Thank you very much for your comment.

We had change ‘y” to “year” in the table.

6. 1 Legends: Please provide the full form of all abbreviations used in the figure for ease of reading.

Answer: Thank you very much for your comment.  

We had provided the full form of all abbreviations in the footnote of Figure 1. “Figure 1. The soil organic (a1,a2), total nitrogen (b1,b2) content and C/N (c1,c2) in vermicompost and soil under different land use types. (the soil in different land use descript as Buttonwood forest (WS), Apple orchard (GS), Walnut forest (TS), Ryegrass land(CS), Kiwi orchard(MS), and Corn field(YS); the vermicompost in different land use descript as Buttonwood forest (WF), Apple orchard(GF), Walnut forest(TF), Ryegrass land(CF), Kiwi orchard(MF), and Corn field(YF))”

7. Table 3: I am curious to know how the authors identified diversity. Since they did not isolate and sequence the species, they only did metagenomics of the soil.

Answer: Thank you very much for your comment.  

We added the identified diversity methods in part 2. As following:

 Alpha diversity—the diversity within a particular region or ecosystem is a comprehensive indicator of richness and uniformity. To comprehensively evaluate the alpha diversity of farmland soil microbial communities, we evaluated the Chao1 index as a representative measure of richness and the Shannon and Simpson indices as representative measures of diversity. Microbiological data processing was conducted using the Parsenor Gene Cloud data analysis platform (http://www.gene scloud.cn), and alpha diversity (Chao1 and Shannon) was calculated.

8.     There are several inconsistencies in font, spacing, and grammatical errors. Please check carefully.

Answer: Thank you very much for your comment.

        We have checked and corrected the mistakes in font, spacing, and grammatical errors

Minor editing of the English language is required as there are some inconsistencies in font, spacing, and grammatical errors.

 

 

 

 

 

 

 

 

3#

Dear editor,

    We have finished modification of my manuscript by your advices.

(I) Please check that all references are relevant to the contents of the manuscript.

Answer: Thank you very much for your comment.

We have checked all references in manuscript.

 

(II) Any revisions to the manuscript should be highlighted, such that any

changes can be easily reviewed by editors and reviewers.

Answer: We had highlighted the revisions in the manuscript

 

(III) Please provide a cover letter to explain, point by point, the details of the revisions to the manuscript and your responses to the referees’ comments.

We have uploaded a cover letter to explain, point by point, the details of the revisions to the manuscript and my responses to the referees’ comments.

 

(IV) If the reviewer(s) recommended references, please critically analyze them to ensure that their inclusion would enhance your manuscript. If you believe these references are unnecessary, you should not include them.

Answer: Thank you very much for your suggestions.

 

(V) If you found it impossible to address certain comments in the review reports, please include an explanation in your appeal.

Answer: Thank you very much for your suggestions.

 

(VI) The revised version will be sent to the editors and reviewers.

Answer: Thank you.