Reviewer’s report 
Title:  Inoculation with enterococci does not affect colon  inflammation in the multi-drug 
resistance 1a-deficient mouse model of IBD 
 
Version:  0 Date:  25 Sep 2015 
 
Reviewer : Yan Lam 
 
Reviewer's report: 
Barnett et al investigated the effect of a single d ose of oral bacterial inoculation on colonic 
inflammation in Mdr1a -/- mice. The authors reporte d that the inoculation was without effect on 
the severity or variability of inflammation. 
Major comments: 
1. From the Introduction I got the impression that the aim of the current study was to develop 
and/or improve an animal model of intestinal inflam mation to study IBD in humans. It 
appeared to me that this manuscript might be more s uitable for a journal that specializes in 
publishing methodology-focused studies. 
2. Please comment on the applicability of the bacte rial inoculation. In the current study the 
inoculation was a mixture of Enterococcus species a nd bacteria from conventionally raised 
mice (assuming these animals were maintained in-hou se). Even if the inoculation were to 
be effective in inducing inflammation in rodents, c an the authors please comment on the 
reproducibility of such an effect? It has been well -established that gut microbiota profile 
varies considerably across laboratories even the an imals are on similar diets, in fact gut 
bacterial compositions could also alter across batc hes of animals that have been maintained 
on the same protocol in the same laboratory.  
3. What I found lacking in the current manuscript w as supporting data on the possible 
molecular mechanisms of the pro-inflammatory effect  of the inoculation in the IL-10 
deficient mice, and the lack thereof, in the curren t study. This would be critical to advance 
our knowledge on the interactions between the immun e system and gut microbiota on 
intestinal inflammation. 
 
 
Are the methods appropriate and well described? 
If not, please specify what is required in your com ments to the authors. 
Yes 
Does the work include the necessary controls? 
If not, please specify which controls are required in your comments to the authors. 
Yes 
Are the conclusions drawn adequately supported by t he data shown? 
If not, please explain in your comments to the auth ors. 
Yes 
Are you able to assess any statistics in the manusc ript or would you recommend an 
additional statistical review? 
If an additional statistical review is recommended,  please specify what aspects require further 
assessment in your comments to the editors. 
I am able to assess the statistics 
Quality of written English 
Please indicate the quality of language in the manu script: 
Acceptable 
Declaration of competing interests 
Please complete a declaration of competing interest s, considering the following questions: 
1. Have you in the past five years received reimbur sements, fees, funding, or salary from an 
organisation that may in any way gain or lose finan cially from the publication of this 
manuscript, either now or in the future? 
2. Do you hold any stocks or shares in an organisat ion that may in any way gain or lose 
financially from the publication of this manuscript , either now or in the future? 
3. Do you hold or are you currently applying for an y patents relating to the content of the 
manuscript? 
4. Have you received reimbursements, fees, funding,  or salary from an organization that holds 
or has applied for patents relating to the content of the manuscript? 
5. Do you have any other financial competing intere sts? 
6. Do you have any non-financial competing interest s in relation to this paper? 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below. If your reply is yes to any, please give det ails below. 
I declare that I have no competing interests. 
I agree to the open peer review policy of the journ al. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named report 
including any attachments I upload will be posted o n the website along with the authors' 
responses. I agree for my report to be made availab le under an Open Access Creative Commons 
CC-BY license (http://creativecommons.org/licenses/ by/4.0/). I understand that any comments 
which I do not wish to be included in my named repo rt can be included as confidential comments 
to the editors, which will not be published. 
I agree to the open peer review policy of the journ al 
Reviewer’s report 
Title:  Inoculation with enterococci does not affect colon  inflammation in the multi-drug 
resistance 1a-deficient mouse model of IBD 
 
Version:  0 Date:  25 Sep 2015 
 
Reviewer : Yan Lam 
 
Reviewer's report: 
Barnett et al investigated the effect of a single d ose of oral bacterial inoculation on colonic 
inflammation in Mdr1a -/- mice. The authors reporte d that the inoculation was without effect on 
the severity or variability of inflammation. 
Major comments: 
1. From the Introduction I got the impression that the aim of the current study was to develop 
and/or improve an animal model of intestinal inflam mation to study IBD in humans. It 
appeared to me that this manuscript might be more s uitable for a journal that specializes in 
publishing methodology-focused studies. 
2. Please comment on the applicability of the bacte rial inoculation. In the current study the 
inoculation was a mixture of Enterococcus species a nd bacteria from conventionally raised 
mice (assuming these animals were maintained in-hou se). Even if the inoculation were to 
be effective in inducing inflammation in rodents, c an the authors please comment on the 
reproducibility of such an effect? It has been well -established that gut microbiota profile 
varies considerably across laboratories even the an imals are on similar diets, in fact gut 
bacterial compositions could also alter across batc hes of animals that have been maintained 
on the same protocol in the same laboratory.  
3. What I found lacking in the current manuscript w as supporting data on the possible 
molecular mechanisms of the pro-inflammatory effect  of the inoculation in the IL-10 
deficient mice, and the lack thereof, in the curren t study. This would be critical to advance 
our knowledge on the interactions between the immun e system and gut microbiota on 
intestinal inflammation. 
 
 
Are the methods appropriate and well described? 
If not, please specify what is required in your com ments to the authors. 
Yes 
Does the work include the necessary controls? 
If not, please specify which controls are required in your comments to the authors. 
Yes 
Are the conclusions drawn adequately supported by t he data shown? 
If not, please explain in your comments to the auth ors. 
Yes 
Are you able to assess any statistics in the manusc ript or would you recommend an 
additional statistical review? 
If an additional statistical review is recommended,  please specify what aspects require further 
assessment in your comments to the editors. 
I am able to assess the statistics 
Quality of written English 
Please indicate the quality of language in the manu script: 
Acceptable 
Declaration of competing interests 
Please complete a declaration of competing interest s, considering the following questions: 
1. Have you in the past five years received reimbur sements, fees, funding, or salary from an 
organisation that may in any way gain or lose finan cially from the publication of this 
manuscript, either now or in the future? 
2. Do you hold any stocks or shares in an organisat ion that may in any way gain or lose 
financially from the publication of this manuscript , either now or in the future? 
3. Do you hold or are you currently applying for an y patents relating to the content of the 
manuscript? 
4. Have you received reimbursements, fees, funding,  or salary from an organization that holds 
or has applied for patents relating to the content of the manuscript? 
5. Do you have any other financial competing intere sts? 
6. Do you have any non-financial competing interest s in relation to this paper? 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below. If your reply is yes to any, please give det ails below. 
I declare that I have no competing interests. 
I agree to the open peer review policy of the journ al. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named report 
including any attachments I upload will be posted o n the website along with the authors' 
responses. I agree for my report to be made availab le under an Open Access Creative Commons 
CC-BY license (http://creativecommons.org/licenses/ by/4.0/). I understand that any comments 
which I do not wish to be included in my named repo rt can be included as confidential comments 
to the editors, which will not be published. 
I agree to the open peer review policy of the journ al 
Please review this paper.
user
Reviewer’s report 
Title:  Inoculation with enterococci does not affect colon  inflammation in the multi-drug 
resistance 1a-deficient mouse model of IBD 
 
Version:  0 Date:  25 Sep 2015 
 
Reviewer : Yan Lam 
 
Reviewer's report: 
Barnett et al investigated the effect of a single d ose of oral bacterial inoculation on colonic 
inflammation in Mdr1a -/- mice. The authors reporte d that the inoculation was without effect on 
the severity or variability of inflammation. 
Major comments: 
1. From the Introduction I got the impression that the aim of the current study was to develop 
and/or improve an animal model of intestinal inflam mation to study IBD in humans. It 
appeared to me that this manuscript might be more s uitable for a journal that specializes in 
publishing methodology-focused studies. 
2. Please comment on the applicability of the bacte rial inoculation. In the current study the 
inoculation was a mixture of Enterococcus species a nd bacteria from conventionally raised 
mice (assuming these animals were maintained in-hou se). Even if the inoculation were to 
be effective in inducing inflammation in rodents, c an the authors please comment on the 
reproducibility of such an effect? It has been well -established that gut microbiota profile 
varies considerably across laboratories even the an imals are on similar diets, in fact gut 
bacterial compositions could also alter across batc hes of animals that have been maintained 
on the same protocol in the same laboratory.  
3. What I found lacking in the current manuscript w as supporting data on the possible 
molecular mechanisms of the pro-inflammatory effect  of the inoculation in the IL-10 
deficient mice, and the lack thereof, in the curren t study. This would be critical to advance 
our knowledge on the interactions between the immun e system and gut microbiota on 
intestinal inflammation. 
 
 
Are the methods appropriate and well described? 
If not, please specify what is required in your com ments to the authors. 
Yes 
Does the work include the necessary controls? 
If not, please specify which controls are required in your comments to the authors. 
Yes 
Are the conclusions drawn adequately supported by t he data shown? 
If not, please explain in your comments to the auth ors. 
Yes 
Are you able to assess any statistics in the manusc ript or would you recommend an 
additional statistical review? 
If an additional statistical review is recommended,  please specify what aspects require further 
assessment in your comments to the editors. 
I am able to assess the statistics 
Quality of written English 
Please indicate the quality of language in the manu script: 
Acceptable 
Declaration of competing interests 
Please complete a declaration of competing interest s, considering the following questions: 
1. Have you in the past five years received reimbur sements, fees, funding, or salary from an 
organisation that may in any way gain or lose finan cially from the publication of this 
manuscript, either now or in the future? 
2. Do you hold any stocks or shares in an organisat ion that may in any way gain or lose 
financially from the publication of this manuscript , either now or in the future? 
3. Do you hold or are you currently applying for an y patents relating to the content of the 
manuscript? 
4. Have you received reimbursements, fees, funding,  or salary from an organization that holds 
or has applied for patents relating to the content of the manuscript? 
5. Do you have any other financial competing intere sts? 
6. Do you have any non-financial competing interest s in relation to this paper? 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below. If your reply is yes to any, please give det ails below. 
I declare that I have no competing interests. 
I agree to the open peer review policy of the journ al. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named report 
including any attachments I upload will be posted o n the website along with the authors' 
responses. I agree for my report to be made availab le under an Open Access Creative Commons 
CC-BY license (http://creativecommons.org/licenses/ by/4.0/). I understand that any comments 
which I do not wish to be included in my named repo rt can be included as confidential comments 
to the editors, which will not be published. 
I agree to the open peer review policy of the journ al 
assistant