Wine Microbial Consortium: Seasonal Sources and Vectors Linking Vineyard and Winery Environments

Round 1

Reviewer 1 Report

This manuscript describes wine microbial consortium and incidence in relation to vineyard environment and sampling season. This study provides a useful database for understanding the parameters affecting wine quality. Basically, the manuscript is well written and presented clearly.

To be clear:

Line 230: All PCR products were sequenced.

Does this mean that 1424 yeast and bacterial isolates were DNA sequenced (line 235)? However, far fewer strains were identified according to Tables 2-5.

Author Response

This manuscript describes wine microbial consortium and incidence in relation to vineyard environment and sampling season. This study provides a useful database for understanding the parameters affecting wine quality. Basically, the manuscript is well written and presented clearly.

• We are grateful for this comment supporting our research work.

Question 1

Line 230: All PCR products were sequenced.

Does this mean that 1424 yeast and bacterial isolates were DNA sequenced (line 235)? However, far fewer strains were identified according to Tables 2-5.

• The phrase is not clear. Only the presumptive lactic acid and acetic acid bacteria as indicated by the respective PCR band were sequenced. This explains the small number of strains identified and reported in tables 2-5. The phrase in line 230 was changed accordingly hoping to be clear now.

Reviewer 2 Report

This manuscript describes the microbial community of a vineyard environment.

Comments:

Line 23: “Gluconoacetobacter spp., Gluconoacetobacter spp.” Repetition, maybe it was “Gluconobacter spp., Gluconoacetobacter spp.”.

Line 34: terroir in italics

Line 26: “Saccharomyces cerevisiae was only isolated….” this sounds strange…. Goddard and colleagues (https://pubmed.ncbi.nlm.nih.gov/19691498/) sampled soil, bark, and flowers from Matua Valley (Auckland) vineyard and identified 122 colonies of S. cerevisiae, with 22 different genotypes. Moreover, Nadai and colleagues (https://www.mdpi.com/2311-5637/5/3/62) reported the isolation of S. cerevisiae from vine bark of an 8 years old vineyard at early spring, late spring and harvest, regardless of the presence of the grape bunch.

Authors should consider these papers and their results when discussing the data….

Line 43: lactic acid, not malic acid

Line 75: Samples

Paragraph 2.1.1: In general, few sites of samplings has been selected, in a restricted area, and it seems that a few number of samples has been collected. For example, only 5 soil samples, and only 1 of them from the forest area far from the vineyard areas. No information is given about the number of bark samples. No information is given about the number of insects sampled and on their species. Only 5 leaves were collected. No information is given about the number of samples collected in the winery. Only 1 sample from grape juices (…at least 3 for a statistic data).

Moreover, no information is given about the vineyard management practices (biological, conventional, …). Different pesticides have different effects on microorganisms and insects and influence their presence.

Figure 1: add the W letter (winery building) in panels a, b and c

Line 172: why only 3 to 5 colonies and not all the colonies with different morphologies?

Line 186: provide NSM medium recipe or add a reference

Paragraph 2.2.2: Did you verify that the DNA you used amplified and was not degraded? Is it possible that you did not have amplification because the DNA was not good and not because the microorganism was not what you were looking for?

Line 248-249: Isn't it also possible that the insects present at different times of the year are different?

Paragraph 3.1: Add a Table with the data for soil / trees / grapes / bark / etc at the different sampling points to be able to compare them (the raw data you use to get the percentages). For example, how many microorganisms were recovered from the soil in winter and how many of them were bacteria, etc.

Remove Figure 2, it is not necessary, the expected band is explained in the text

Remove Figure 3, it is not necessary, the expected band is explained in the text

Line 301: “eight samples” how many from tree bark and how many from insects?

Line 301: contaminated is not correct, please use another term

Line 303: Debaryomyces hansenii and D. robertesiae in italics

Line 303: change later with latter

Line 305: Candida cellae, C. parapsilopsis, Metschnikowia spp. and Metschnikowia chrysoperlae in italics

Line 307: Debaryomyces spp., Metschnikowia spp., and Candida spp.. D. hansenii and D. fabryi in italics

Line 309: C. albicans, C. hawaiiana, C. apicola and M. rekaufii in italics

Line 310-311: What's the point of sampling the winery if the equipment was new?

Line 312-315: C. hawaiiana - H. uvarum, H. optuntiae and C. infanticola - Pichia ciferrii, P. guilliermondii and H. uvarum in italics

Line 317: H. uvarum and P. guilliermondii in italics

Line 318: P. guilliermondii in italics

Line 322-324: Metschnikowia spp. (M. pulcherrima), Candida spp. (C. zemplinina, C. diversa, C. helenica), Issatchenkia spp. (I. terricola), Pichia spp. (P. fermentans), and Hanseniaspora spp. (H. uvarum) in italics

Line 325: P. manshurica in italics

Line 327: S. cerevisiae, Z. bailii, and P. occidentalis in italics

Line 336: contaminated is not correct, please use another term

Paragraph: 3.3.2 and 3.3.3: Again, genus and species in italics!!!

Line 350 is Paragraph 3.3.3 and not 3.3.2

Due to these comments, it is recommended that this manuscript should not be published in its current form. 

Author Response

Please see the attachment

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

Ok