On 2017 Jul 26, Martine Crasnier-Mednansky commented:

I do appreciate your answer to my comment, to which I gladly reply. First, there is prior work by Ghosh S, 2011 indicating colonization was attenuated in mutant strains that were incapable of utilizing GlcNAc, which included a nagE mutant strain. Second, Mondal M, 2014 analyzed the products of the ChiA2 reaction and found GlcNAc was the most abundant product. In fact, the amount of (GlcNAc)2 was found to be very low as compared to GlcNAc and (GlcNAc)3. Therefore, it is fully legitimate to conclude the PTS substrate GlcNAc is utilized in the host by V. cholerae for growth and survival.

On 2017 Jul 26, Ankur Dalia commented:

Re: Martine Crasnier-Mednansky

I appreciate your evaluation of the manuscript, however, I have to disagree with your comment. The study by Mondal et al. indicates that ChiA2 can liberate GlcNAc from mucin in vitro and that it is critical for bacterial growth in vivo, however, they did not test the role for GlcNAc uptake and/or catabolism in that study. In our manuscript, however, we demonstrate that loss of all PTS transporters (including the GlcNAc transporter) does not result in attenuation in the same infant mouse model, which is a more formal test for the role of GlcNAc transport during infection. It is formally possible that other carbohydrate moieties are liberated via the action of ChiA2 that are required for growth of V. cholerae in vivo, however, our results would indicate that these are not translocated by the PTS. Alternatively, the reduced virulence of the ChiA2 mutant observed in the Mondal et al study may indicate that ChiA2 has other effects in vivo (i.e. on immune evasion, resistance to antimicrobial peptides, etc.).

On 2017 Jul 24, Martine Crasnier-Mednansky commented:

The authors’ proposal 'the PTS has a limited role during infection' and concluding remark 'PTS carbohydrates are not available and/or not utilized in the host' are both questionable. Mondal M, 2014 established, when Vibrio cholerae colonizes the intestinal mucus, the PTS-substrate GlcNAc is utilized for growth and survival in the host intestine (upon mucin hydrolysis).

On 2017 Jul 24, Martine Crasnier-Mednansky commented:

The authors’ proposal 'the PTS has a limited role during infection' and concluding remark 'PTS carbohydrates are not available and/or not utilized in the host' are both questionable. Mondal M, 2014 established, when Vibrio cholerae colonizes the intestinal mucus, the PTS-substrate GlcNAc is utilized for growth and survival in the host intestine (upon mucin hydrolysis).

On 2017 Jul 26, Ankur Dalia commented:

Re: Martine Crasnier-Mednansky

I appreciate your evaluation of the manuscript, however, I have to disagree with your comment. The study by Mondal et al. indicates that ChiA2 can liberate GlcNAc from mucin in vitro and that it is critical for bacterial growth in vivo, however, they did not test the role for GlcNAc uptake and/or catabolism in that study. In our manuscript, however, we demonstrate that loss of all PTS transporters (including the GlcNAc transporter) does not result in attenuation in the same infant mouse model, which is a more formal test for the role of GlcNAc transport during infection. It is formally possible that other carbohydrate moieties are liberated via the action of ChiA2 that are required for growth of V. cholerae in vivo, however, our results would indicate that these are not translocated by the PTS. Alternatively, the reduced virulence of the ChiA2 mutant observed in the Mondal et al study may indicate that ChiA2 has other effects in vivo (i.e. on immune evasion, resistance to antimicrobial peptides, etc.).

On 2017 Jul 26, Martine Crasnier-Mednansky commented:

I do appreciate your answer to my comment, to which I gladly reply. First, there is prior work by Ghosh S, 2011 indicating colonization was attenuated in mutant strains that were incapable of utilizing GlcNAc, which included a nagE mutant strain. Second, Mondal M, 2014 analyzed the products of the ChiA2 reaction and found GlcNAc was the most abundant product. In fact, the amount of (GlcNAc)2 was found to be very low as compared to GlcNAc and (GlcNAc)3. Therefore, it is fully legitimate to conclude the PTS substrate GlcNAc is utilized in the host by V. cholerae for growth and survival.