Experimental Long-Term Conservation of an Infant-Sized Piglet—Efficacy of a Late 17th Century Embalming Procedure

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

This manuscript presents a carefully executed and unusually long-term experimental reconstruction of a late seventeenth-century dry embalming protocol. The seven-year follow-up, combined with macroscopic assessment, serial histology, and whole-body CT imaging, represents a rare degree of methodological patience and technical consistency. The work is clearly the result of sustained commitment, and it makes a meaningful contribution to experimental paleopathology and the study of historical preservation techniques. With several refinements, the manuscript can be strengthened further in terms of positioning, interpretation, and methodological transparency.

First, the historical and conceptual framing would benefit from a sharper articulation of the specific knowledge gap the study addresses. The introduction provides solid background on embalming traditions and correctly identifies Penicher’s protocol as one of the few sources offering detailed quantitative instructions. However, the manuscript would be strengthened by more explicitly stating why replicability enabled by Penicher’s precise measurements is scientifically decisive in this context. In addition, the link between the experimental model and previously observed infant mummies should be framed more cautiously and analytically. At present, the rationale moves from historical observation to experimental reconstruction in a logical but somewhat compressed manner. Clarifying whether the aim is to test plausibility, reproduce preservation patterns, or assess long-term structural stability would sharpen the conceptual focus.

Second, the issue of sample size deserves clearer acknowledgment. While a single experimental model is understandable given the duration and labor intensity of the protocol, the manuscript should explicitly frame the study as an experimental demonstration or proof-of-concept rather than as a validation of efficacy in a broader sense. The current language occasionally implies the general effectiveness of the Penicher protocol. It would be more precise to state that substantial long-term preservation was demonstrated in this specific experimental case. A brief but explicit discussion of generalizability—particularly regarding environmental conditions, animal-to-human anatomical differences, and variability in historical ingredient quality—would improve the methodological rigor of the discussion.

Third, the macroscopic evaluation scoring system should be contextualized more critically. Although the semi-quantitative approach is reasonable for longitudinal observation, its subjective nature should be acknowledged. Clarifying whether assessments were performed by a single observer or multiple evaluators, and whether any attempt at standardization or reproducibility was made, would enhance transparency. Even a short statement recognizing the inherent subjectivity of this scale would suffice.

The ethical section would also benefit from minor clarification. The manuscript states that no additional regulatory approval was required under EU law because the animal had been euthanized for unrelated medical reasons. While this may indeed be correct, it would strengthen the paper to specify whether institutional confirmation or formal documentation was obtained. A brief clarification that the work involved post-mortem tissue only, without experimental intervention on a living animal, would preempt potential concerns.

In the Results and Discussion sections, the internal organ loss is a particularly interesting finding and could be explored in greater depth. The disappearance of residual internal structures despite the absence of overt tissue destruction raises mechanistic questions. Was this primarily due to dehydration, microbial autolysis before complete desiccation, or insufficient penetration of preservative substances into deeper cavities? A short analytical reflection on why skin and muscle were preserved while internal organs were not would add conceptual depth and move the paper beyond descriptive reporting.

Similarly, the persistence of epidermal nuclear remnants for several years is striking and merits slightly more interpretative commentary. Is this consistent with known patterns of desiccation-mediated preservation? A brief comparison with ancient mummy histology literature could strengthen the interpretative layer of the manuscript.

The conclusion should also be moderated slightly in tone. Statements describing the protocol as “very successful and effective” may be softened to reflect the single-model design. A formulation emphasizing substantial long-term preservation under controlled conditions would be more proportionate to the presented data.

Finally, although the English language is generally clear and technically sound, minor stylistic refinements could improve precision. Some sentences in the Discussion section are lengthy and could be tightened to improve flow. These are editorial rather than substantive issues and do not impede comprehension.

In summary, this is an original and carefully conducted experimental reconstruction study with unusually long follow-up and commendable methodological detail. The manuscript would benefit from clearer positioning of its scientific scope, more explicit acknowledgment of limitations (particularly regarding sample size and generalizability), and slightly moderated claims in the concluding section. With these refinements, the study will stand as a valuable contribution to experimental investigations of historical embalming techniques and long-term tissue preservation.

Author Response

Please see the attachement

Author Response File: Author Response.docx

Reviewer 2 Report

Comments and Suggestions for Authors

I read and reviewed the manuscript titled Experimental long-term conservation of an infant-sized piglet – efficacy of a late 17th century embalming procedure by Nerlich et al.
While the paper reports a fascinating experiment aimed at testing an embalming protocol from the past, I don’t believe it should be given the relevance of a full article but rather a case report. After all, only one sample was embalmed and analyzed, seems only fair to me to treat it as such. Beside this initial consideration I think there are some things that need to be addressed before moving to the acceptance of the manuscript in the journal Anatomia.

Below, I will breakdown some of the critical points I’ve detected in the manuscript:

 

Title page

Line 4-13: Make sure that formatting is according to the template of the journal throughout the manuscript. For instance, here, the numbers for affiliations should be given in superscript in the author list. The institutions specifications are given in a font size that is too small to read clearly.

 

Introduction

Line 59: Refer it as Supplementary materials, not supplement.

 

Materials and Methods

Lines 122-204: This part sounds more like text from an alchemy book rather than a scientific paper, please add the scientific names to all the herbs/seeds used to prepare the embalming mixture and use the international system to refer measures, change cubits and fingers to the appropriate measures in cm or m. Whilst this part is surely fascinating, I believe it could be given as Supplementary information rather than in the full text of the manuscript.

Lines 206-251: This could go in the Supplementary material as well.

Line 282: Add information on the scale used and on the accuracy of the measurement, did you measure to the nearest gram or with less accuracy?

Line 283: Was the evaluation carried out by the same observer throughout the period of 7.2 years? Specify how you reduced inter-observer bias. Was this scale reported before in literature or you came up with it? What was the rationale behind it? Did you expect to observe those changes overtime?

Table 1: Correct “funghi” to “fungi”.

 

Results

Line 313-317: This is a paragraph for the Discussion section, you’re not stating results here.

Figure 2: I believe that this figure should be completely redone. I am having issue in understanding what does the y axis represent. Did you transform the categorical scoring in table one in a continuous variable? Besides, numbers in the x axis does not correspond to the bars in the graph.

Figure 3: Again, add labels on the two axes.

Line 420: Remove this line.

 

Discussion

Lines 422-448: This section mirrors what you have already stated in the introduction, consider removing it from the discussion.

 

Make sure that the references in the reference sections are formatted with the referencing typesetting requested by the journal

Author Response

Please see the attachement

Author Response File: Author Response.docx

Reviewer 3 Report

Comments and Suggestions for Authors

The authors describe an experimental protocol of embalming procedure used in the past. In particular, they adopt the protocol suggested by two authors in the 17th centuries.  

The manuscritp appear interesting over all for museum technicians and exhibition speciman preparers. It appear well written and documented. The references are apropritaed. The images of good quality are important to better understand the preparation processes. The methods well described and the results riched by different methods: macroscopical observations, microscopical sections (the choice of colors was suitable to highlight the various structures). 

I think that the unique things that could be revised are the facts that: for this experiment has been used only one piglet. It should be more valid having used three animals. However, I understand that this work started 8 years ago and this long time limits the experimantal dectetion of results. So I think that it can be left as it is. 

In figure 4 I suggest to rotate the imgae of 180° as the vertebrae must stay in dorsal position. 

In final consideration, I thing that this manuscript deserves to be published because it is of interest for museum experts.   

Author Response

Please see the attachement

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The revised version of the manuscript has adequately addressed all major concerns raised in the previous review. The authors have significantly improved the clarity of the conceptual framework, expanded the methodological explanations, and strengthened the discussion, particularly regarding limitations and interpretation of the findings.

The manuscript is now scientifically sound, well-structured, and clearly written. It represents a valuable contribution to the field and is suitable for publication in its current form.

Author Response

We thank the reviewer again for his very helpful comments and suggestions and are happy to hear that he now recommends the publication of our paper.

Reviewer 2 Report

Comments and Suggestions for Authors

Thank you for the changes made in the manuscript. It was rather difficult to follow everything because the manuscript file was not in tracked changes, and you did not provide line numbering in the response letter.
I strongly suggest you doing it for your next reviews, it is easier for everyone and it is also timesaving for reviewers.
There are a couple things that still need to be done.
- Table 1 must be given using the table layout provided by the journal;
- Same goes for the subchapter partitioning and the formatting of figure captions, check what the instructions for author say;
- remove the "." at line 417;
- change "supplementum" to "supplementary materials" at line 494.
I have no other remarks

Author Response

We are again very grateful to this reviewer for his helpful suggestions. In detail we made following changes:

Table I was reformatted according to the journal´s recommendation, the table has been brought into font 8 and has now replaced the previous version.

Subchapter partitioning was adopted to the journal´s instructions by numbers.

The "" was removed (now line 420).

The term "supplementum" was replaced by "supplementary materials" as suggested.