Composition of the Wild Rabbit Nest and Its Implication for Domestic Rabbit Breeding
Round 1
Reviewer 1 Report
Please see the attached document.
Comments for author File: 
Comments.docx
Author Response
L60-62: This sentence expresses a personal opinion and should be placed in the discussion instead of the introduction
Response: this section has been deleted based on another reviewer's comment.
L 71: please specify the weight of the rabbits used (there are 2 types of chinchilla rabbits, a light and a heavy weight one). This is important for the assessment of the cage sizes used in the experiment.
Response: mean body weight was added.
L72: Please specify where your laboratory is.
Response: location was given
L73: If these are “standard” cages please specify the housing guidelines according to which you kept the rabbits. It should be at least according to the EU Directive 2010/63, Annex III B, Table 2.2.
Response: the word statndard has been deleted because the cage was substantially larger than a standard cage
L73 & 74: I guess the measurements refer to length x width x height? Please specify.
Response: length, width and height measurements were specified
L76: I presume L:D stands for Light:Dark? Please spell out all abbreviations used when they first appear in the text.
Response: abbreviations were replaced by Light and Dark respectively
L79: I miss a reference to the DIRECTIVE 2010/63/EU OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 22 September 2010 on the protection of animals used for scientific purposes
Response: at the end of the manuscript the permission of this trial is presented
L 90: Please explain, how you determined the dryness and the length of the material of the wild rabbit nests.
Reponse: the requested information was added
L100: Did you determine the dryness of your artificially dried gras? If yes, did it match the results of the wild nests? If no, why not? And what happened after you took the nests away? Taking a nest away without replacement is very stressful for the does and might put the animal welfare at risk. This needs to be taken into consideration!
Response: this section was rephrased (nest material were replaced by the same amount of hay)
L104-105: The behavioural observation needs to be explained in much more detail. Which observation hour did you choose? Why did you observe the behavior for one hour only? Usually, longer time periods are observed (at least 24 hours) and the observations are repleated over a couple of days. Did you use direct or video observation? Please explain, why you used the chosen method. Which method was used? Time sampling, continuous recording? Behaviour sampling? Please provide an ethogram, with the exact definitions of the recored behaviours. Martin and Batesons standard book for scientific behaviour observation (“measuring behaviour”) might be useful.
Response: the used method is specified
L107-109: I don’t really understand this part of the Experiment. In experiment 2 you know whether the does used the dry or the fresh grass (by measuring how much was taken out of each of the tussocks or by observing the rabbits using it (this should be made clear in the M&M section, too!). I do not understand, why you stored these nests for 4 weeks.
Responese: the composiition of materials by the does was analyzed in the way described above for wild rabbit nest.
L109 where do the “ten nests of known initial composition” come from? Experiment 2 used 7 does only. Did you repeat the experiment? If so, you forgot to mention this. This section is very confusing to me.
Response: the nest composition was examined for 7 nest (text has been modified accordingly). The experiment was not repeated.
L115: Why did you not test the commonly used wood shavings, too? This would have been very interesting.
Response: the environmental conditions of the European Wild rabbit was followed where wood shavings are not available.
L121-123: See comments to behaviour observation in L104-105
Response: the used method is specified
Table 1 and 2: The formatting of this table is confusing. Please spell out the abbreviation “SE” when first using it. Why is the specification of the plant material important for this project? Did you make use the same plants in Experimemt 1 and 2? If yes, it is not mentioned. If not, this information seems somewhat out of place. Maybe you should include the plant species in the introduction and discussion sections, to warrant such a detailed table in the results section. If you decide to keep the details of the plants in the table, I suggest using the full scientific names as in the text.
Response: Standard error waswritten. According to our opinion providing genus is sufficient.
L155ff: The material and methods section does not explain, how you came to these results. You refer to behaviours like “exploration”, “hopping”, “sniffing” “eating” etc. There needs to be a definition in the MM section exactly how (method) and which (definitions) behavior was recorded.
Response: the main focus of this study was examining nest building behaviour rather than other behaviour forms, therefore this part will not be extended
Figure 1: The y-axis Time % of what? Please provide more precise axis labels. Please define “collected”
Response: clarification was provided
Figure 2: There are no letters indicating significant differences. The description of the microscopic observation belongs into the MM section.
Response: letters were added
Figure 3: There is no asterix. To which time does the percentage in the x-axis refer to? You should mention the total time observed in the figure caption.
Response: asterics were added
L208-216: I am sure, there is more literature to support this paraghraph. As a general rule in scientific writing, all literature used in the introduction section should be cited again in the discussion section, putting your own findings into perspective. Even though this rule doesn’t need to be followed 100% - it should be at least a guideline. You only refer to very few citations mentioned in the introduction in the discussion!
Response: this section was completely rephrased
L217: Which data? In the paragraph above you refer to general biological traits of rabbits and not wo any specific (scientific) data.
Response: this section was completely rephrased
L208-223: The first 2 paragraphs of the discussion do not seem to be related to the research topic. They should be either shortened or the relation to the research topic (nest material) should be made clearer.
Response: this section was completely rephrased
L233-234: Really? The data in your results do not support this. To support this conclusion, you should present data showing the percentage of different grass species in the national park, where you found the nests and examine, whether the grasses found in the park and in the nests are of the same composition.
Response: this section was completely rephrased
241-242: I don’t understand this sentence. What do you want to say?
Response: this section was completely rephrased
L245: Why did you not weigh or determine the volume taken out of the tussocks? This would have made it easy to determine how much from each sort of gras was taken. Why did you not specify which sort of grass you used on the laboratory experiments, as the type of grass is an important factor in experiment 1?
Response: in order to keep the same methodology as used in Experiment 1
Besides, the same species were used that were identified in the nests of the European Wild Rabbits
Ll268-269: I understood that you could not differentiate between eating dry and green gras in the nest box due to darkness within the nest box? There are no results to support this hypothesis presented in the results section.
Response: this section was completely rephrased
L288: As mentioned above, please specify where these “standard conditions” are written down. In my opinion they should at least fulfill the requirements of the EU Directive 2010/63. Depending on the weight of the rabbits used and the not quite clear measurements (it is not clear what is length, width and height) it remains doubtful to the reader whether this was the case. You mention, that you reduced the animals for the experiment to the minimum, but you do not explain how you came to a number of only 7 does per experiment in the statistical methods section.
Response: length, width and hight values were specified. The word standard was deleted as the used cages were larger than required by the EU directive.
Using a small sample size still enabled us to find large scale differences among the groups.
Discussion in General: I miss a critical material and methods discussion. Your results refer to 1 experiment with 7 dams and 1 hour of observation per experiment (2 and 3) only. This is a very small dataset to generalize any of the results.
Response: this section was
References: Even though I appreciate, that you have done a thorough research many decades back, it is striking, that you use very old literature. It puzzles me that only 10 out of 46 references were published in the new millennium, which is not so new anymore, after all. Only 2 references are from the last 5 years. I recommend checking this. If there is a good explanation, why there is hardly any current literature on your topic, this should be mentioned in the discussion section.
Response: the manuscript was extended by adding recent and relevant references.
Reviewer 2 Report
Review comments
This study was undertaken to understand the factors affecting the nest material choice of rabbits by analysing the composition of wild rabbit nests and testing naive domestic does in controlled laboratory settings for their preferences in constructing their nests. The authors collected 21 just abandoned nests in their natural habitat and found they contained rabbit fur and hay, of which 85 % was dry grass, mostly long leaves of Calamagrostis sp.. The authors’ domesticated rabbit does we then tested if there is a preference for the features of the wild rabbit nests or whether the above-mentioned finding was either an accidental consequence of grass availability or ageing of the nest material. Two-way choice tests revealed that primiparous domestic does preferred the dry grass over green and long leaves over short grass bunches for nest building. The authors concluded that long dry grass instead of short wood shavings is recommended as a nest material for domesticated rabbits.
For major comments plz see these points and for details see the attached file.
- The novelty of the work should be cited in the introduction section.
- References in the introduction section and discussion can be updated with
- The asterisks and letters mentioned in the footnotes of the figures are absent
- The taken home massage should be mentioned in the abstract and summary of the MS.
- In the discussion section authors may gave some attention for the basis of rabbits’ behavior for collect of the grass for nest building and feeding.
All in all, the Ms deserve publication after major revision. It meets the journal readership and have both scientific and practical merits.
Comments for author File: 
Comments.pdf
Author Response
First of all, authors would like to thank the reviwer's effort improving our mansucript with comments and suggestions. We are going to respond the raised points as follows.
The novelty of the work should be cited in the introduction section.
Response: the requested section has been added.
References in the introduction section and discussion can be updated with
Response: the references were updated with more recent articles
The asterisks and letters mentioned in the footnotes of the figures are absent
Response: the letters and asterisks are provided
The taken home massage should be mentioned in the abstract and summary of the MS.
Response: the take home message is added at the end of the abstract
In the discussion section authors may gave some attention for the basis of rabbits’ behavior for collect of the grass for nest building and feeding.
Response: the discussion section has been completely re-written.
Besides, the reference that was suggested by the reviwer has been added.
Reviewer 3 Report
GENERAL COMMENT:
I consider this work is within the scope of “Applied Sciences”. It contains information useful in a field in which available information is scarce and of interest to improve knowledge on nest making and maternal behaviour of rabbits. Overall, experimental design is correct and the results have been adequately discussed. However, I indicate several suggestions to be considered by the authors to improve the manuscript. I indicate these recommendations below and in a commented PDF file I have uploaded.
HEADING (heading): Indicate manuscript type: “Article”
CITATION (first page):
Please fill this part with authors' names and manuscript title.
ABSTRACT:
Line 9: Insert “(Oryctolagus cuniculus)” after “rabbits”, resulting in “Wild European rabbits (Oryctolagus cuniculus)”.
Line 14: Add: “·...or straw”; there are also rabbit farmers that use straw for the nest.
Line 25-26: As you have not compared wood shavings with the other materials in your experiments, I consider “We concluded that long dry grass instead of short wood shavings is recommended as a nest material for domesticated rabbits” is not a proper conclusion. I suggest modifying it to link the conclusion more clearly to your results.
KEYWORDS:
Add: Oryctolagus cuniculus.
INTRODUCTION:
Line 32: Insert "from predators", thus resulting in “hide the litter from predators”.
Line 36: According to the stile of the journal for the citation or references, replace [3,4,5,6,7,8] with [3-8].
Lines 37-39: Provide a bibliographic reference to support the following sentence: “Physical features like insulation capacity of the nest are of utmost importance where outer conditions are harsh and energetic costs are high”.
Line 52: Replace with "kits". "Pups" is a word not recommended by the World Rabbit Science Association.
Lines 57-58: This has also been demonstrated for wild rabbits in captive condition. See: González-Redondo, P. (2010). Maternal behaviour in peripartum influences preweaning kit mortality in cage-bred wild rabbits. World Rabbit Science, 18(2), 91-102.
Line 59: In addition to laboratory conditions it is advisable to refer to farming conditions, where straw is also used (not only wood shavings).
MATERIALS AND METHODS:
Line 73: Write “ad libitum” in italics.
Line 74: Due to the fact that the nest box characteristics influence the nest making by the doe, and taking into account that exist different nest box designs (closed with a lid or open, without upper lid), it is necessary a more precise description of the nest box (closed or open in the upper part, size and position of the entrance for the doe, etc.).
Line 77-78: This law is no longer in force from 2013, as you can see at: https://eur-lex.europa.eu/legal-content/EN/ALL/?uri=CELEX%3A31986L0609
Provide the proper regulation now in force (or in force when the experiment was carried out).
Line 130: Rewrite in a better way; for example: “IBM SPSS Statistics 27.0 (Student License) was used to perform the statistical analyses.”
RESULTS:
Line 138: Indicate what is this dispersion statistic: SD, SE?
DISCUSSION:
This section is correctly addressed and organised.
REFERENCES SECTION:
In general terms, this section adjust to style and format of the journal.
TABLES:
Table 2: Write Latin scientific names in italics.
FIGURES:
Figure 1: Asterisks are lacking.
Figure 2: Letters above the colums are lacking.
Figure 3: Asterisks are lacking.
Comments for author File: Comments.pdf
Author Response
The authors are grateful for receiving the suggestions and comments of the reviewer. The raised points are answered as follows:
HEADING (heading): Indicate manuscript type: “Article”
Response: indicated
CITATION (first page):
Response: the requested information was given
ABSTRACT:
Line 9: Insert “(Oryctolagus cuniculus)” after “rabbits”, resulting in “Wild European rabbits (Oryctolagus cuniculus)”.
Response: “(Oryctolagus cuniculus)” after “rabbits”, has been inserted
Line 14: Add: “·...or straw”; there are also rabbit farmers that use straw for the nest.
Response: the suggested modification has been performed
Line 25-26: As you have not compared wood shavings with the other materials in your experiments, I consider “We concluded that long dry grass instead of short wood shavings is recommended as a nest material for domesticated rabbits” is not a proper conclusion. I suggest modifying it to link the conclusion more clearly to your results.
Response: the conclusion section has been modified
KEYWORDS:
Add: Oryctolagus cuniculus.
Response: the suggested key word is added
INTRODUCTION:
Line 32: Insert "from predators", thus resulting in “hide the litter from predators”.
Response: "from predators" has been inserted
Line 36: According to the stile of the journal for the citation or references, replace [3,4,5,6,7,8] with [3-8].
Reponse: the suggested format has been adapted
Lines 37-39: Provide a bibliographic reference to support the following sentence: “Physical features like insulation capacity of the nest are of utmost importance where outer conditions are harsh and energetic costs are high”.
Response: a related reference is added
Line 52: Replace with "kits". "Pups" is a word not recommended by the World Rabbit Science Association.
Response: pups are relpaced by kits
Lines 57-58: This has also been demonstrated for wild rabbits in captive condition. See: González-Redondo, P. (2010). Maternal behaviour in peripartum influences preweaning kit mortality in cage-bred wild rabbits. World Rabbit Science, 18(2), 91-102.
Response: the suggested references are inserted
Line 59: In addition to laboratory conditions it is advisable to refer to farming conditions, where straw is also used (not only wood shavings).
Response: the suggestion has been adapted
MATERIALS AND METHODS:
Line 73: Write “ad libitum” in italics.
Response: italic was used
Line 74: Due to the fact that the nest box characteristics influence the nest making by the doe, and taking into account that exist different nest box designs (closed with a lid or open, without upper lid), it is necessary a more precise description of the nest box (closed or open in the upper part, size and position of the entrance for the doe, etc.).
Response: more detailed information was given
Line 77-78: This law is no longer in force from 2013, as you can see at: https://eur-lex.europa.eu/legal-content/EN/ALL/?uri=CELEX%3A31986L0609
Provide the proper regulation now in force (or in force when the experiment was carried out).
Repsonse: the proper regulation was given and the number of our permission performing the present study was also added
Line 130: Rewrite in a better way; for example: “IBM SPSS Statistics 27.0 (Student License) was used to perform the statistical analyses.”
Response: modified as suggested
RESULTS:
Line 138: Indicate what is this dispersion statistic: SD, SE?
Response: Standar error
DISCUSSION:
This section is correctly addressed and organised.
REFERENCES SECTION:
In general terms, this section adjust to style and format of the journal.
TABLES:
Table 2: Write Latin scientific names in italics.
Response: itallics is adapted
FIGURES:
Figure 1: Asterisks are lacking.
Response: asterisks are added
Figure 2: Letters above the colums are lacking.
Response: letters were added
Figure 3: Asterisks are lacking.
Response: asterisks are provided
Reviewer 4 Report
The presented paper deals with an original topic, which seems to be important because the behaviour of rabbit does is a key fuctor for understanding challenges in commercial breedings or in smallholder´s conditions. In my opinion, the results of the study are inovative and interesting due to their connection with study of wild rabbit does´ nests with overlap to commercial conditions.
My minor comments or suggestions:
L85: How did you collect the nest material? Into some sterile bags?
L90: Please, provide the mark of the manufacturer of the microscope.
L96: How old were the females? Did you choose them randomly?
L154: In Experiment 2, I suggest to unify using of % or percent.
Author Response
Authors highly appreciate the valuable comments and suggestions of the reviewer. Please find our responses below.
L85: How did you collect the nest material? Into some sterile bags?
Response: nest material was collected into sealable steril bags
L90: Please, provide the mark of the manufacturer of the microscope.
Response: NIKON SMZ1270
L96: How old were the females? Did you choose them randomly?
Response: 30 weeks old randomly chosen rabbits
L154: In Experiment 2, I suggest to unify using of % or percent.
Response: % was used
Round 2
Reviewer 1 Report
Dear authors,
thank you for revising the manuscript. Even if the overall dataset is still thin, the the intersting topic of your study warrants publication.
Reviewer 2 Report
The authors responded good to reviewers comments and addressed all issues
