Fungal Community Analyses of a Pirogue from the Tang Dynasty in the National Maritime Museum of China
Round 1
Reviewer 1 Report
Lines 2-3: reduce line spacing.
In the title please change “the pirogue” by “a pirogue from the Tang Dynasty” and “in China” by “of China”.
Lines 14-15: remove.
The paper has some errors and inconsistencies (mainly related to English language), which need to be fully addressed. I suggest a native English speaker's proofreading.
Key question: what is the specific application or potential application of the study? It is not clear. Some experiments have been performed but what was the practical objective towards an implementation.
Line 34: include “of China”.
Line 38: “The wood has been identified as Rhodamnia dumetorum”. Please include Reference/s.
Lines 41-46: The works dealing with the presence of fungi and bacteria in wood heritage are common in scientific literature. These few examples introduced in this way are not understood. Please give an overview of the problem, focusing later on these examples. Make it clear also in what regions of China these examples are.
Line 61: but for what purpose?
I would include the Figure S1 in the paper, as Figure 1. It is relevant.
Lines 105 and 127: change “by Zijun Liu [2].” by “by Liu et al. [2].”
Letters in Figure 1 must be increased, they are illegible.
Table 1 must appear on a single page.
The quality of Figure 3 must be improved. Letters are illegible.
Lines 138-139: chang “by Ronglin He [18].” by “by He et al [18].”
Figure 3 caption and Figure 3 must appear on a single page.
Another key question: what is the biodeterioration and in what extent in the pirogue?
Overall, in my opinion, the study is merely descriptive and presents only a microbial community analysis. In addition to the two key questions (already mentioned) that have not been answered in the paper, the study with biocides is not clear nor the practical protocol to follow for the conservation of the pirogue in the museum.
This reference should be included and also some of the references therein:
Sanmartín, P.; DeAraujo, A.; Vasanthakumar, A. (2018) Melding the old with the new: trends in methods used to identify, monitor and control microorganisms on cultural heritage materials. Microbial Ecology 76: 64-80.
Author Response
Please see the attachment
Author Response File: 
Author Response.pdf
Reviewer 2 Report
In the reviewed article, the Authors applied combined culture-dependent and independent methods to asses microbial community involved in biodeterioration process of canoe. The Authors used also four biocides to analyze sensitivity of microbial strains occurring on the canoe surface. Manuscript requires major revision and remarks listed below entail further explanation. Language correction by native speaker will add additional value to the manuscript.
Abstract
Line 22: What about bacterial community? What bacterial genera were predominant?
Line 23: Please add information about type of four biocides or active substances used in this study?
Introduction
Introduction should include more information on biodeterioration process of wood and artifacts made from this material.
Line 53: Please modify phrase “fungal corrosion”, because corrosion is a process regarding usually destruction of metal surfaces by chemical and/or electrochemical reaction with the environment.
Line 59: What means fungal population in this case? Maybe fungal strains will be more appropriate in this context.
Materials and methods
Materials and Methods section is too general and requires detailed description.
Line 63: In my opinion chapter “Sample collection” does not contain a description of the sampling methods, but the characteristics of the object (canoe) and should be modified.
Line 72: Please delete “microbial damage”
Line 78: The Authors should be more precise about the sampling methods, area, part of canoe etc. Please add these information.
Line 84: Why did the Authors determine only fungal contamination in the air?
Line 93: Did the Authors isolate and identify bacteria or fungi, please explain, because it is unclear.
Line 100: Please add information, which method was used for evaluation of DNA purity and concentration?
Line 102: fungal sequences? Please modify this sentence.
Lines 106-107: Please complete the methodology of the gene libraries construction, purification and sequencing methods, or provide appropriate literature.
Lines 109-117: The chapter “Bioinformatic analysis” lacks details, what may be confusing for the readers. Which software, or databases did the Authors use for bioinformatic analysis of high-throughput sequencing data? Please specify and add more details.
Line 127: no spaces
Line 132: 1×108 spores per ml, please specify the unit
Line 147: Could the Authors specify, which biocides and active compounds were used for the experiment? It would be interesting to know, what concentrations range the Authors used in experiments?
Line 147: Did the cultivation of fungal strains perform only 2 days?
Results
In my opinion this chapter needs ordered. For me it is still unclear and difficult to understand, whether the Authors analyzed bacteria or fungi or maybe both groups of microorganisms.
Line 152: Why in the title did the Authors focus on identification of fungi, while the results also include bacteria. Why “main fungi”? In high-throughput sequencing analysis culturable and nonculturable microorganisms were detected not only predominant. Please modify the title.
Line 179, Figure 1: high-throughput sequencing should be with a lowercase letter
Line 184: Please modify “had different predominant genera”
Line 199: I appreciate the effort put in the fungi identification, however I am wondering why some isolates were identified to the species level, while some of them to the genus? I strongly recommend to apply molecular techniques based not only on ITS region, but also calmodulin, tubulin, actin genes sequencing as an additional method, which may be helpful in complete identification.
Line 210, Figure 2: This figure is not required, because it does not provide new information.
Line 265: no space
Line 274: Please see comment aforementioned in line 147. Information on biocides and active compounds is necessary.
Discussion
The Authors focused again on description the results, that’s why discussion needs to be provided in the details.
Author Response
Please see the attachment
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
The manuscript in this new version is certainly improved thanks to the authors’ effort to fulfil the modifications requested.
In my opinion, paper in its current form is suitable for publication in Applied Sciences.
Author Response
Thank you for your comments concerning our manuscript.
Reviewer 2 Report
Dear Authors,
In the reviewed article entitled “Fungal community analyses of a pirogue from the Tang Dynasty in the National Maritime Museum of China”, changes to my comments have been prepared in a sufficient way. In my opinion, the manuscript requires only some minor changes (listed below).
Introduction
Line 51: “fungal mycelia” should be with a lowercase letter.
Lines 76-77: Please, modify the sentence: “Traditional methods…” as follows: “Traditional methods based on cultivation and microscopy techniques are expensive and time consuming”
Line 79: Should be “has been used…”
Line 81: “traditional” should be with a lowercase letter.
Materials and methods
Line 103: Please, change “viable” to “visible”
Line 143: Please change “amplicon sequencing” to “high-throughput sequencing”
Line 151: White space was inserted (°C ,), please remove.
Line 167: Should be “were prepared as described Liu et al.” In References, Liu et al. is 6 or 7 position, not 2. Please check the citations throughout manuscript.
Line 168-169: Please modify as follow: “Finally, the PCR products were used for library construction and sequencing were performed on the Illumina HiSeq2500 PE250 platform”
Lines 178 and 188: lack of the white space
Line 206: Should be “One ml” instead of “A 1 ml”
Results
Please enter full species names before strain numbers e.g. line 354: “Penicillium pimiteouiense NKDMZ-1” instead of “NKDMZ-1 (Penicillium sp.)” throughout manuscript.
Figure 2: Figure 2 should be bigger, because is not readable. Why the relative abundance does not reach value 1 (100%) on the Figure 2B? Please check it.
Discussion
Line 398 and 402: Please change “Amplicon sequencing” to “high-throughput sequencing”
Line 403: Cryptococcus, and Aspergillus
Line 434: Please remove Keiko
Line 436: Should be Klavker et al. not Katja et al.
Line 452: Aspergillus, Penicillium and Cladosporium should be in italics
Line 452: Borrego et al. lack of dot
References
Please check the references carefully, you often write journals names with lowercase.
Author Response
Please see the attachment
Author Response File: Author Response.pdf
