Biodeterioration Assessment of a Unique Old Pharaonic Kingdom Wooden Statue Using Advanced Diagnostic Techniques
, Shimaa Ibrahim 2, Chiaramaria Stani 3, Giovanni Birarda 4, Nehal Ali 5, Emam Abdullah 6, Lisa Vaccari 4, Paola Grenni 7,*, Andrea Visca 7, Yehia Badr 8 and Wafaa Soliman 8
Cosmin Spîrchez
Round 1
Reviewer 1 Report
The subject is presented clearly. It is necessary to complete with 5 bibliographical references from 2021 year.
Author Response
Response to Reviewer 1 Comments
Point 1: The subject is presented clearly. It is necessary to complete with 5 bibliographical references from 2021 year
Response 1 Thank you for your suggestion. The following references were added in the text:
Branysova, T.; Demnerova, K.; Durovic, M.; Stiborova, H. Microbial biodeterioration of cultural heritage and identification of the active agents over the last two decades. J. Cult. Herit. 2022, 55, 245–260, doi:10.1016/j.culher.2022.03.013.
Pyzik, A.; Ciuchcinski, K.; Dziurzynski, M.; Dziewit, L. The Bad and the Good—Microorganisms in Cultural Heritage Environments—An Update on Biodeterioration and Biotreatment Approaches. Materials (Basel). 2021, 14, 177, doi:10.3390/ma14010177.
Microorganisms in the Deterioration and Preservation of Cultural Heritage; Joseph, E., Ed.; Springer International Publishing: Cham, 2021; ISBN 978-3-030-69410-4.
Zisi, A. Forest Wood through the Eyes of a Cultural Conservator. Forests 2021, 12, 1001, doi:10.3390/f12081001.
Liu, X.; Qian, Y.; Wang, Y.; Wu, F.; Wang, W.; Gu, J.-D. Innovative approaches for the processes involved in microbial biodeterioration of cultural heritage materials. Curr. Opin. Biotechnol. 2022, 75, 102716, doi:10.1016/j.copbio.2022.102716.
Reviewer 2 Report
The conclusions and techniques are meaning for the selection of appropriate methods for their consolidation and recovery treatments. However, there are some minor unclear problems needing revisions before it could be accepted for publication.
1. The information of water in Archaeological wooden objects is suggested to be added in the introduction section.
2. Line 118. km2, the number of 2 should be marked using superscript.
3. Line 158. The photos in figure 2 are not clear enough, replace them if possible.
4. Line 174. The moisture content of samples is recommended to be presented in the table.
5. ‘Three dimensional plans, transvers, longitudinal and radial?’
Generally, wood has three distinguished planes for observation in tangential, radial and transverse.
6. Line 189. The moisture content of wood samples for the test need to control or not for the Microscopic and SEM-EDX observations? These questions are also relevant to other measurements or observations; please describe the moisture content information of materials used in the article if needed.
7. Summarize more concise and clear conclusions as possible.
Author Response
Response to Reviewer 2 Comments
The conclusions and techniques are meaning for the selection of appropriate methods for their consolidation and recovery treatments. However, there are some minor unclear problems needing revisions before it could be accepted for publication.
- The information of water in Archaeological wooden objects is suggested to be added in the introduction section.
Response:
Thank you for you observation and suggestion. We have added more information on this aspect in the statue description. Really, at the moment of the statue discovery, the wood moisture was not measure. In fact, this measure requires sampling of wood to be weight and to be put in an oven. In order to avoid any damage the statue, this was not carried out. Since the discovery until today, the statue has been kept in the best air humidity conditions for its conservation and analyses (in fact, wet analysis is not ideally suited for examining archaeological wooden objects, as also stated by some authors (YoonSoo Kim, Adya P. Singh, Wood as Cultural Heritage Material and its Deterioration by Biotic and Abiotic Agents, in: Editor(s): Yoon Soo Kim, Ryo Funada, Adya P. Singh, Secondary Xylem Biology, Academic Press, 2016, Pages 233-257).
- Line 118. km2, the number of 2 should be marked using superscript.
Response: Ok, done
- Line 158. The photos in figure 2 are not clear enough, replace them if possible.
Response: Thank you for your comment. We have added the new Figure 2 with a higher resolution, as also required by the Reviewer 4.
- Line 174. The moisture content of samples is recommended to be presented in the table.
Response: Thank you for your comment. As reported above, we have not registered the moisture of wood samples; this information did not influence the results.
- ‘Three dimensional plans, transvers, longitudinal and radial?’
Generally, wood has three distinguished planes for observation in tangential, radial and transverse.
Response: Thank you for your suggestion. We have corrected as follows: three dimensions (transversal, tangential and radial sections).
- Line 189. The moisture content of wood samples for the test need to control or not for the Microscopic and SEM-EDX observations? These questions are also relevant to other measurements or observations; please describe the moisture content information of materials used in the article if needed.
Response: Thank you for your comment. The Microscopic and SEM-EDX observations do not require the information on wood moisture and this parameter does not affect the measurement outcomes.
- Summarize more concise and clear conclusions as possible.
Response: Thank you. The conclusions are now more concise.
Reviewer 3 Report
This manuscript is a novel and interesting study. However, a few comments need to be addressed before publication.
1. Abstract focused too much on material and method but provided too brief info about results and discussion. I recommend that the authors rewrote the abstract with an emphasis on discussion and conclusion.
2. The keywords preferably should not come from the title of the paper.
3. More literature should be added to the introduction. For example, considering the title of the paper, references can be added related to fungal decay.
Bari, E., Daryaei, M. G., Karim, M., Bahmani, M., Schmidt, O., Woodward, S., ... & Sistani, A. (2019). Decay of Carpinus betulus wood by Trametes Versicolor-An anatomical and chemical study. International Biodeterioration & Biodegradation, 137, 68-77.
Schmidt, O., Bahmani, M., Koch, G., Potsch, T., & Brandt, K. (2016). Study of the fungal decay of oil palm wood using TEM and UV techniques. International biodeterioration & biodegradation, 111, 37-44.
4. In discussion, the results of the current research should be compared with previous studies.
5. Conclusion is too long. Authors are encouraged to summarize the conclusion and tailor it to the results of the current study. Besides, research limitations and recommendations for future studies should be provided.
Author Response
This manuscript is a novel and interesting study. However, a few comments need to be addressed before publication.
- Abstract focused too much on material and method but provided too brief info about results and discussion. I recommend that the authors rewrote the abstract with an emphasis on discussion and conclusion.
Response: Thank you for you very useful comment. The abstract was revised. We hope now it is clearer.
- The keywords preferably should not come from the title of the paper.
Response: Thank you for your comment. The new keywords are the following:
micro-FT-IR; Fluorescent in situ hybridization; Weathering effect; Wood Deterioration; Crystallinity of cellulose; Abu-Rawash statue; burial soil analyses
- More literature should be added to the introduction. For example, considering the title of the paper, references can be added related to fungal decay.
Bari, E., Daryaei, M. G., Karim, M., Bahmani, M., Schmidt, O., Woodward, S., ... & Sistani, A. (2019). Decay of Carpinus betulus wood by Trametes Versicolor-An anatomical and chemical study. International Biodeterioration & Biodegradation, 137, 68-77.
Schmidt, O., Bahmani, M., Koch, G., Potsch, T., & Brandt, K. (2016). Study of the fungal decay of oil palm wood using TEM and UV techniques. International biodeterioration & biodegradation, 111, 37-44.
Response: Thank you for your comment. We have added some recent references as required by the reviewer 1 and we have also added the papers you suggested.
- In discussion, the results of the current research should be compared with previous studies.
Response: Thank you for your suggestion. We have improved the discussions as follows:
Page 12
This spectral range showed a broadening and a decrease in the infrared features definition that can be linked to a decrease in the structural order of the wooden tissues in the archae-ological woods, if compared to the standard one. This was already observed by several authors [30,67]. (Burattini & Falcieri, 2020; Y. Kim & Singh, 2016).
Page 13
As expected, in Figure 8a the false-colour FPA images of the Acacia standard sample showed the highest amounts (corresponding to the red colour in the maps and the scale bar) of cellulose and lignin (Fig. 8b, orange and green rectangles, respectively) and the lower amount of deterioration products (Fig. 8a, blue rectangle). FPA map of sample #1 for cellulose still showed a good preservation and a wide distribution with just some degraded regions (blue spots), while the lignin content was quite low with just some preserved pale red spots [13]. (Singh et al., 2022).
Moreover, the references of the Discussion section were improved.
- Conclusion is too long. Authors are encouraged to summarize the conclusion and tailor it to the results of the current study. Besides, research limitations and recommendations for future studies should be provided.
Response: Thank you. The conclusions are now more concise, as you requested.
Reviewer 4 Report
The manuscript is dealing with the characterization of an ancient wooden statue from Egypt. The paper has a logical structure, however, it is lacking on some information and quality of the figures used. Overall, I suggest major revisions, please see the details below.
Image quality of Figure 2 is very low. Please replace it by using good quality images.
Line 165: Please clear the meaning of "transvers, longitudinal and radial". Main anatomycal directions of wood are transversal, radial and tangential. How is it related to your sampling directions?
Why did you use Acacia nilotica as a standard wood sample? The manuscript did not contain any result about the identification of the statues wood species. Thus, it is not supported to use a standard wood sample of a certain wood species. Please support the use of Acacia nilotica by wood anatomical identification of the statue. Without this, you must delete the results related to the standard sample.
Why you used sapwood sample, when the sample from the statue was from the inner part of it? Inner part of the statue must be heartwood.
Lines 313-316: Please add a reference to support this assumption.
Figure 6a-b: As it is a comparison, please use images with the same characteristics. Please include images with the same magnifications. Now you are using 600× and 800× magnifications, that is not acceptable in this case. But the much bigger problem with these images is, that Fig. 6a shows a cross section, while Fig. 6b shows a longitudinal section. This is totally unacceptable for showing any feature, change or difference between the two samples. Please modify the figure by using imiges of same section and same magnification.
Figure 6a-b: Furthermore, description of part c) is missing from the image title.
Figure 6d-e-f: To be honest, the image quality is very low here. Thus, it is not possible tosee the necessary details to identify the deterioration of the ancient samples, or the structure of the standard sample. I cannot recognize any wood anatomical feature in Fig. 6d neither. Furthermore and here again, you are using different magnification for each image, while you are comparing them. Please replace images of Fig. 6d-e-f by a good quality images using the same magnification for all.
Line 374: you wrote: ". d) Average spec...", but this is obviously about section g) of the image.
Figure 7a: You show on the right of Fig. 7a the degradation rate (0-100%). But this image also containing the lignin and cellulose rates, using the same colors (red and blue). With this layout it mens that lignin and cellulose have a degradation rate of 100% in case of the standard sample, while nearly 0% degradation rate in case of the ancient samples. Of course, it is totally false, but with the used layout the image has this meaning. Please modify the lignin and cellulose part of Fig. 7a to fix this problem. Add separated scale bar for cellulose and lignin rates.
Line 577: please correct the word "wooed"
Line 581: please correct this part: "allow as to"
Author Response
Response to Reviewer 1 Comments
The manuscript is dealing with the characterization of an ancient wooden statue from Egypt. The paper has a logical structure, however, it is lacking on some information and quality of the figures used. Overall, I suggest major revisions, please see the details below.
Image quality of Figure 2 is very low. Please replace it by using good quality images.
Response: Thank you for your comment. We have added the new Figure 2 with a better resolution.
Line 165: Please clear the meaning of "transvers, longitudinal and radial". Main anatomycal directions of wood are transversal, radial and tangential. How is it related to your sampling directions?
Response: Thank you for your suggestion. We have corrected as follows:
“three dimensions (transversal, tangential and radial sections)"
Why did you use Acacia nilotica as a standard wood sample? The manuscript did not contain any result about the identification of the statues wood species. Thus, it is not supported to use a standard wood sample of a certain wood species. Please support the use of Acacia nilotica by wood anatomical identification of the statue. Without this, you must delete the results related to the standard sample.
Response: Thank you for your comment. Section “2.3 Sample description and wood classification” reports the wood classification method used. Moreover, we have moved to the Result section the results on the wood identification (that was previously incorrectly present in the M&M).
Why you used sapwood sample, when the sample from the statue was from the inner part of it? Inner part of the statue must be heartwood.
Response: Sorry, you are right. The right Table 1 is the following:
|
Samples |
Description |
Sampling position |
Colour |
|
#1 |
Pure core wooden sample |
Inner the statues |
Dark Brown |
|
#2 |
Wood interfered with soil |
Inner the statues |
Light Brown |
|
#3 |
Standard sample (Acacia nilotica) |
- |
Fresh wood colour |
|
#4 |
Soil sample |
Inner the statue |
Red |
|
#5 |
Soil sample |
Petrified outer layer |
White |
|
#6 |
Soil sample |
Inner the statue |
Dark grey |
Lines 313-316: Please add a reference to support this assumption.
Response: Thank you for your suggestion. We have added a reference as follows:
SEM-EDX spectrum (Figure 3d, wood sample #1) shows the presence of very high amount of Ca, Cl and Si and lower amounts of Na, Mg, Al, S, K, Pb and Fe; this might be attributed to the penetration of soil components inside the wood, as these elements are the main components of soil (Volkov et al., 2021).
Volkov, D., Rogova, O., & Proskurnin, M. (2021). Organic Matter and Mineral Composition of Silicate Soils: FTIR Comparison Study by Photoacoustic, Diffuse Reflectance, and Attenuated Total Reflection Modalities. Agronomy, 11(9), 1879. https://doi.org/10.3390/agronomy11091879
Figure 6a-b: As it is a comparison, please use images with the same characteristics. Please include images with the same magnifications. Now you are using 600× and 800× magnifications, that is not acceptable in this case. But the much bigger problem with these images is, that Fig. 6a shows a cross section, while Fig. 6b shows a longitudinal section. This is totally unacceptable for showing any feature, change or difference between the two samples. Please modify the figure by using imiges of same section and same magnification.
Response: Thank you for your suggestion. For a more legibility of the two figures, Fig. 6 a and b are now a separate figure. Moreover, we have changed the text as follows:
SEM image in Figure 6a is a transverse section of Acacia nilotica standard sample. This diffuse-porous wood is characterized by solitary or multiples vessels. Axial parenchyma was vasicentric, aliform lozenge-aliform, and confluent where the homogeneous structure of wood cell walls clearly appeared. On the contrary, the ancient wood sample #1 (Figure 6b) showed an advanced decay, in which a wide wood fragmentation is visible. Cell walls look collapsed, cracked vessels and voids are clearly observable. This leads to a porosity and fragility increase of the wood structure. Such wood cells separations might be attributed to the collapse of middle lamella region that leads to reduce the homogeneity of wood structure causing either fracture or evacuation and cell wall erosion (Zidan et al., 2016).
Figure 6a-b: Furthermore, description of part c) is missing from the image title.
Response: Thank you for your comment. In the new figure 7 this description is now added.
Figure 6d-e-f: To be honest, the image quality is very low here. Thus, it is not possible to see the necessary details to identify the deterioration of the ancient samples, or the structure of the standard sample. I cannot recognize any wood anatomical feature in Fig. 6d neither. Furthermore and here again, you are using different magnification for each image, while you are comparing them. Please replace images of Fig. 6d-e-f by a good quality images using the same magnification for all.
Response: Thank you for your suggestion. We have added figures 4 and 7 with a higher quality. Moreover, the structure of the samples, including the wood anatomical feature, are not clearly recognizably due to the sample preparation (sample compression in the diamond cell), as also explained in Materials and Methods and in the text page 12. We have also put the images with the same magnification.
Line 374: you wrote: ". d) Average spec...", but this is obviously about section g) of the image.
Response: Thank you, yes you are right. Now the figure legend is corrected.
Figure 7a: You show on the right of Fig. 7a the degradation rate (0-100%). But this image also containing the lignin and cellulose rates, using the same colors (red and blue). With this layout it mens that lignin and cellulose have a degradation rate of 100% in case of the standard sample, while nearly 0% degradation rate in case of the ancient samples. Of course, it is totally false, but with the used layout the image has this meaning. Please modify the lignin and cellulose part of Fig. 7a to fix this problem. Add separated scale bar for cellulose and lignin rates.
Response: Yes, you are right, sorry, the misunderstanding of the old Figure 7, now Figure 8, was because the blue-red scale of the figure on the left (a) was too much close to the y axis of the figure (b) on the right. The blue-red scale of the figure on the left is now put in the right place to avoid confusion.
Line 577: please correct the word "wooed"
Response: Thank you for your suggestion. Done
Line 581: please correct this part: "allow as to"
Response: Thank you for your suggestion. Done
Reviewer 5 Report
The article presents the results on the mechanisms of biodeterioration of ancient wooden statue caused by chemical- and biological processes.
In general, work was performed using appropriate methods and the results obtained support the conclusion. Thus, the article may be accepted after minor revisions:
1. The analysis of microbial population was carried out using culture-based technique and further analysis of enrichments with FISG and qPCR. Thus, it is better to make separate sub-section “Microbial population analysis” with sub-sub-section: “Isolation of microorganisms”, “FISH analysis”, “qPCR analysis”.
2. Nutrient medium selection should be explained, as the analysis based on culture-depended technique depends on the medium used.
Author Response
The article presents the results on the mechanisms of biodeterioration of ancient wooden statue caused by chemical- and biological processes.
In general, work was performed using appropriate methods and the results obtained support the conclusion. Thus, the article may be accepted after minor revisions:
- The analysis of microbial population was carried out using culture-based technique and further analysis of enrichments with FISG and qPCR. Thus, it is better to make separate sub-section “Microbial population analysis” with sub-sub-section: “Isolation of microorganisms”, “FISH analysis”, “qPCR analysis”.
Response: Thank you for your observation. Really when the tapes were applied to the wood surface, a very few number of microbial cell was observed under the Epifluorescence microscope, and they were not enough to perform FISH technique and PCR analyses. Consequently, FISH was performed on microbiological samples taken from archaeological wooden samples using adhesive tapes immediately placed in a cultivation broth. We have explain better this in the "2.7 Microbial analyses by Fluorescent In Situ Hybridization (FISH)" section of M&M and we hope that now it is clearer.
- Nutrient medium selection should be explained, as the analysis based on culture-depended technique depends on the medium used.
Response: Thank you for your suggestion. Several additional information are now added in the M&M section
Reviewer 6 Report
The investigation is professionally implemented and rigorously reported.
Some abbreviations are used without giving the full meaning, which requires detective work from the interested reader.
It appears that some references are quoted in terms of author names, instead of numbers in the order of appearance.
This reviewer has difficulties in understanding the writing style used in the Discussion.
In addition to factors related to wood deterioration, the Discussion contains statements regarding ancient societies. This reviewer feels such statements possibly should be more carefully expressed.
Author Response
The investigation is professionally implemented and rigorously reported.
Some abbreviations are used without giving the full meaning, which requires detective work from the interested reader.
Response: Thank you for your suggestion. We have added the full meaning of several abbreviations, as follows:
PCR (polymerase chain reaction) (in the abstract and introduction sections)
Fourier-transform infrared (FT-IR) spectroscopy (in the introduction section)
Gran Egyptian Museum (GEM)
Focal Plane Array (FPA) (in the Fig. 4 caption)
It appears that some references are quoted in terms of author names, instead of numbers in the order of appearance.
Response: Thank you for your observation, the Reference list was corrected
This reviewer has difficulties in understanding the writing style used in the Discussion.
Response: Thank you for your comment. The Discussion section was improved, the Authors hope that now it is clearly.
In addition to factors related to wood deterioration, the Discussion contains statements regarding ancient societies. This reviewer feels such statements possibly should be more carefully expressed.
Response: Thank you for your comment. The M&M and the discussion sections were improved for this part.
Round 2
Reviewer 3 Report
The authors have addressed the comments satisfactorily. Therefore, the manuscript in the current version is acceptable for publication.
Reviewer 4 Report
The authors improved the quality according to the suggestions of the reviewers. I suggest to accept the manuscript for publication in its present form.
