Nest Material Preference of Wild Mouse Species in Laboratory Housing
Round 1
Reviewer 1 Report
Overall comment: Throughout the manuscript, the words "grass" and "hay" are used interchangeably, leading to some confusion. Was fresh grass used or hay? I believe it was hay, so clear definition of the type of grass that was used to make the hay, followed by consistent use of the word "hay" would help reduce confusion.
Line 95: Do you know the health status of these mice? Are they screened for any murine pathogens?
Line 98: Why did you choose 52 of each mouse species? Were all of these mice wild caught or were any laboratory born? What was their age (range with mean)?
Line 100: What is a "T4 laboratory mouse box?"
Line 101: How was the water treated?
All Materials and Methods: Please provide source/manufacturer for all materials used (e.g. hay pockets, nesting materials). Strongly recommend review of the ARRIVE guidelines for guidance on what information should be included to assist other researchers in replicating your study.
Line 105: Were all cages provided with 100 to 150 grams of each type of substrate? Why such a wide range? Were they provided mixed in a single hay packet or in three separate hay packets? Were they provided the substrates once or daily? Was grass or hay provided?
Line 108: How does one randomly select strands from finished nests? Would it not have been better to separate the nest and weigh the components?
Lines 155-164: Can you provide the actual data (ranges, probability, etc) for the nesting composition scores?
Author Response
First of all, authors would like to thank the reviewer's evaluation, remarks and suggestion. Our detailed answer is as follows:
Overall comment: Throughout the manuscript, the words "grass" and "hay" are used interchangeably, leading to some confusion. Was fresh grass used or hay? I believe it was hay, so clear definition of the type of grass that was used to make the hay, followed by consistent use of the word "hay" would help reduce confusion.
The suggestion is accepted and the word grass has been replaced by hay throughout the manuscrip.
Type of grass was: Centreal European monocotyledonous species (Versele-laga mountain hay 50 l)
Line 95: Do you know the health status of these mice? Are they screened for any murine pathogens?
The place of our study was a conventional laboratory where there is a permanent health checks are performed by vets, an dthe general hygenie requirements are met. On the contrary no screening is performed for murine pathogens.
Line 98: Why did you choose 52 of each mouse species? Were all of these mice wild caught or were any laboratory born? What was their age (range with mean)?
All the animals were born in the lab since the population is bred since 25 generations (occasionally some animals are received from another laboratories). The age of the animals used in this study ranged between 100 and 120 days.
According to the relevant literature the minimum group size per predictor variable is 10 but the minimum suggested total sample size is 100.
Line 100: What is a "T4 laboratory mouse box?"
T4 laboratory mouse box is a polycarbonate box =600 x 200 x 380 mm) with a basic area of: 1815 cm2 (EU standard)
Line 101: How was the water treated?
The water was supplied ad libitum using nipple drinkers and was filled twice per week
All Materials and Methods: Please provide source/manufacturer for all materials used (e.g. hay pockets, nesting materials). Strongly recommend review of the ARRIVE guidelines for guidance on what information should be included to assist other researchers in replicating your study.
Materials:
Hay: Versele-laga mountain hay 50 l
Cotton: MultiFit small rodent cottob bedding 30l
Paper: Enviro-Dri Rodent Bedding 20 kg
Line 105: Were all cages provided with 100 to 150 grams of each type of substrate? Why such a wide range? Were they provided mixed in a single hay packet or in three separate hay packets? Were they provided the substrates once or daily? Was grass or hay provided?
Within every hay pocket each subtsrate was placed at the first day of the experiment side by side (not mixed). The range was necessary in order to provide all subtsrates in the same volume.Hay was provided.
Line 108: How does one randomly select strands from finished nests? Would it not have been better to separate the nest and weigh the components?
Concerning the nest component determination the method of Szenczi et al. (2011) was followed where after evaluating the quality of the nests they were placed on a tray then every nest was homogenized and subsequently 20 samples were taken blind using forceps.
Lines 155-164: Can you provide the actual data (ranges, probability, etc) for the nesting composition scores?
The composition of each nest was characterized by the 20 samples. The descriptive statistics are as follows:
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Mean |
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Maximum |
Std Error |
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Reviewer 2 Report
Dear Authors,
I appreciated your effort and dedication to the manuscript. However, I have some comments. From my point of view, in the introduction, the subtitle "Nets building in animal kingdom" is not represented by the short lines that you wrote, which mainly point out mice and rabbits. It should be more accurate to highlight the importance of the selection of nest-building material in mice.
Line 27. Remove the first sentence, "Nesting is a common form of behavior..." this allows the reader to be more focused on the paper's relevance
Line 51. Please provide a bibliography to sustain the paragraph, and this is important to make your document more accurate.
I consider that the aim of the study should be clear to the reader. Sometimes, it appears to "the present study examined how wild mouse species choose from natural nest material" (see line 13, abstract). In the introduction, you mention the environmental enrichment of the species related to ecological needs (see lines 85 to 88). It could be more precise and improve the document if you homogenize the objective of the work throughout the drafting of the manuscript.
You described clearly the methods, and your experimental design considered an equal number of males and females of both species. But please, clarify how did you run the experiments simultaneously. How many individuals per group and sex of each species do you use at 10º and 21ºC.
Results. Please consider removing the graphic (Figure 1). In lines 129 and 130, the values are clear enough.
You analyzed the sexes, and you did not find significant differences, except in the choice of cotton (lines 139-141). It will be necessary to include the possible differences between sexes in the introduction and the discussion, including the choice of cotton by females.
Line 157. "The higher amount of hay improves the quality of the nest" is repeated in line 159.
Please consider that if your data point out a preference for a specific material, in your discussion, you should remark because, during the discussion, you only mention choice, which is different in meaning. In fact, nest material preference is the title of your study.
Author Response
We would like to thank the reviewer's efforts in order to improve the scientific quality of our manuscript. After carefully considering the suggestions our detailed response is the following:
I appreciated your effort and dedication to the manuscript. However, I have some comments. From my point of view, in the introduction, the subtitle "Nets building in animal kingdom" is not represented by the short lines that you wrote, which mainly point out mice and rabbits. It should be more accurate to highlight the importance of the selection of nest-building material in mice.
The subtitle has been modified to: small mammals
Line 27. Remove the first sentence, "Nesting is a common form of behavior..." this allows the reader to be more focused on the paper's relevance
The suggestion has been accepted
Line 51. Please provide a bibliography to sustain the paragraph, and this is important to make your document more accurate.
Additional reference was provided
I consider that the aim of the study should be clear to the reader. Sometimes, it appears to "the present study examined how wild mouse species choose from natural nest material" (see line 13, abstract). In the introduction, you mention the environmental enrichment of the species related to ecological needs (see lines 85 to 88). It could be more precise and improve the document if you homogenize the objective of the work throughout the drafting of the manuscript.
The main objective of our study was choosing the nest material for the laboratory mice. Nevertheless it also serves as an environmental enrichment as in many rodent house the animals are kept on wood shavings thus providing natural nest building substrates (helping the manifestation of nest building behaviour forms) also has an important environmental enrichment role.
You described clearly the methods, and your experimental design considered an equal number of males and females of both species. But please, clarify how did you run the experiments simultaneously. How many individuals per group and sex of each species do you use at 10º and 21ºC.
In both rooms 52 animals were individually housed (provided with 3 nest materials).
In the colder and warmer rooms (10 ℃, 21 ℃) 26-26 house mouse and mound-builiding mouse were kept whit an equal sex ratio (13 males and 13 females, respectively).
Results. Please consider removing the graphic (Figure 1). In lines 129 and 130, the values are clear enough.
Figure 1 has been deleted.
You analyzed the sexes, and you did not find significant differences, except in the choice of cotton (lines 139-141). It will be necessary to include the possible differences between sexes in the introduction and the discussion, including the choice of cotton by females.
In the mus genus nests are built by both sexes. Unfortunately no relevant study was found supporting our finding. However, looking the results the preference of cotton was negligible (less than 1 piece from the 20 nest samples) in both sexes, so it was decided that although the difference was significant it was not relevant thus it was not included in the discussion section.
Line 157. "The higher amount of hay improves the quality of the nest" is repeated in line 159.
The suggestion is accepted.
Please consider that if your data point out a preference for a specific material, in your discussion, you should remark because, during the discussion, you only mention choice, which is different in meaning. In fact, nest material preference is the title of your study.
The term choice (which was used once in the discussion section) has been replaced by providing in the last paragraph
Round 2
Reviewer 2 Report
Dear Authors,
I consider that the manuscript changed accordingly. The concepts and the corrections that you made improve the manuscript. The reading in methods is straightforward, and the discussion changes for the better.
Sincelery
