Review Reports

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

First of all, I welcome the efforts of the authorial team to highlight such an interesting and important subject!

The article is devoted to a problem involving both veterinary and public health. Therefore its actuality is out of any doubt.

However, I have some comments that (I hope) would contribute to improving of this manuscript.

The Title is clear and reasonably short to reflect the content.

The Simple Summary can be (but not mandatory) more simple and non-specialist-reader-friendly. Presently it can serve as a professional abstract.

The Abstract contains all the necessary components: background, M&M, results, discussion and conclusion. There are two unexplained abbreviation (PCR and ITS), but they are too well-known in the field to require explanations.  

Keywords: if there is no requirement of alphabetic order, it is alright, otherwise it should be rearranged.

 Introduction. There is a full (but not too long) background regarding both invasive mink in Lithuania and Trichinella. The aim is clearly declared.

Materials and Methods.

Lines 103-104: “Sampling sites were predominantly located in 103 central and eastern Lithuania (Figure 1) and were kept frozen at -20 C until examinations 104 had been conducted.” Something happened to this sentence, because sampling sites could not be kept frozen, whereas samples could. Kindly, improve it.

I suggest adding the time period when meat samples were collected. Was it just 2024?

Coordinated (decimal degrees) of the sampling sites are desirable.

Scale bar and degrees are desirable for Figure 1.

Line 127-128: The purification of DNA was performed according to the method of Pozio and La Rosa 2003 [21], with several modifications. Which ones? Are they described as follows in the text?

What were positive and/or negative controls for DNA-analysis (amplification)? What was an outgroup during phylogenetic analyses?

Results. Line 193: The infected individual exhibited a low larval burden. Is there a document prescribing which amount is considered low? Otherwise it sounds as a value judgment.

Discussion. Is comprehensive enough.

 

Conclusions. Corresponds to the aim and results.

 

References. Are relevant and numerous enough.

 

Overall, the article is very good.

Author Response

Point 1: The Simple Summary can be (but not mandatory) more simple and non-specialist-reader-friendly. Presently it can serve as a professional abstract.

Response 1: Thank you for your coment. We changed the simple summary.

Point 2: The Abstract contains all the necessary components: background, M&M, results, discussion and conclusion. There are two unexplained abbreviation (PCR and ITS), but they are too well-known in the field to require explanations. 

Response 2: Thank you, we added the explanations for the abbreviations.

Point 3: Keywords: if there is no requirement of alphabetic order, it is alright, otherwise it should be rearranged.

Response 3: Thank you. We have changed it.

Point 4: Lines 103-104: “Sampling sites were predominantly located in 103 central and eastern Lithuania (Figure 1) and were kept frozen at -20 C until examinations 104 had been conducted.” Something happened to this sentence, because sampling sites could not be kept frozen, whereas samples could. Kindly, improve it.

Response 4: Thank you. We correct the sentence

Point 5: I suggest adding the time period when meat samples were collected. Was it just 2024?

Response 5: Thank you. Added.

Point 6: Coordinated (decimal degrees) of the sampling sites are desirable.

Response 6: We thank you for this suggestion. However, precise geographic coordinates are not available, as the specimens were obtained from the Kaunas Tadas Ivanauskas Museum of Zoology collection, where such data were not recorded. All available locality information has been included.

Point 7: Scale bar and degrees are desirable for Figure 1.

Response 7: We appreciate this suggestion. A scale bar has been added to Figure 1. However, geographic coordinates could not be included, as precise location data for the sampling sites are not available.

Point 8: Line 127-128: The purification of DNA was performed according to the method of Pozio and La Rosa 2003 [21], with several modifications. Which ones? Are they described as follows in the text? 

Response 8: The modifications are described in detail in the subsequent text. To improve clarity, we have revised the sentence to explicitly indicate that the modified protocol is presented below.

Point 9: What were positive and/or negative controls for DNA-analysis (amplification)? What was an outgroup during phylogenetic analyses?

Response 9:  Thank you, we have added a description of the use of controls in Methods. Phylogenetic trees were not generated during our analysis and for haplotype networks outgroups are not necessary

Point 10: Results. Line 193: The infected individual exhibited a low larval burden. Is there a document prescribing which amount is considered low? Otherwise it sounds as a value judgment.

Response 10: Thank you for this comment. The subjective term “low” has been removed, as there is no universally accepted threshold for classifying larval burden.

Reviewer 2 Report

Comments and Suggestions for Authors

Review of the article "First Report of Trichinella spiralis in Free-Living Invasive American Mink (Neovison vison) in Lithuania" by Maziliauskaitė et al.

The article title and content are consistent with the journal's focus.

1. Simple Summary. I would recommend the authors make this section more understandable to the general reader, as required by the journal's guidelines, by removing the names of genomic regions and Latin names.

2. Abstract. "Trichinella larvae were detected in 5.6% of the animals investigated..." The number must be specified (1 of 18).

3. Introduction.
- Lines 44-56: This paragraph should be shortened and the American Mink population and range in Lithuania should be presented more concisely. Discussion of World War II is unnecessary.
- Lines 57-70: The main idea should also be presented in one sentence. This section needs to be revised, as the main topic is trichinellosis in American mink in Lithuania. It is necessary to analyze and present data on whether mink infections with trichinellosis have been recorded in Europe or other countries, and on which continents.

4. Figure 1. Please indicate in the text or figure caption the program used to create the figure and its source.

5. "Reactions were carried out in a final volume of 25 µL, containing 12.5 µL of DreamTaq PCR Master Mix (Thermo Fisher Scientific, Vilnius, Lithuania), 0.5 µM of each primer, and 4 µL of template DNA." The total volume is 17.5 µL. What was the remaining volume?!

6. Please transfer the sequenced and deposited sequence data (GenBank accession numbers), as well as the reference strain numbers used, to Section 2.4. DNA Sequence Analysis.

7. Figure 2. The figure title needs to be changed; digestion was not artificial, but rather using artificial gastric juice.

8. Line 198. "Ten Trichinella larvae were isolated from the infected animals." You're talking about multiple infected animals here, although line 191 reports one infected animal - "Ten Trichinella larvae were isolated from the infected animals...".

9. The authors report studying 10 larvae, but Figure 2 only shows 5 samples. Explain this.

10. Figure 2. N – negative control. What was used as the negative control? Include this in the figure title or in the description of the PCR method.

11. Lines 220-251. There's too much descriptive text for the Results section; shorten it and move the description to the Discussion section.

12. The Discussion section, in my opinion, is also more descriptive and similar to the Introduction. The authors have divided the Discussion section into subsections. However, this section should contain comparative data obtained by you and other authors. Compare the numerical data, rather than simply listing where and in which countries trichinosis in minks has been detected.

Author Response

Point 1: Simple Summary. I would recommend the authors make this section more understandable to the general reader, as required by the journal's guidelines, by removing the names of genomic regions and Latin names.

Response 1: Thank you for your recommendation. We changed the simple summary.

Point 2: Abstract. "Trichinella larvae were detected in 5.6% of the animals investigated..." The number must be specified (1 of 18).

Response 2: Thank you. We specified.

Point 3: - Lines 44-56: This paragraph should be shortened and the American Mink population and range in Lithuania should be presented more concisely. Discussion of World War II is unnecessary.

Response 3: We have corrected as suggested.

Point 4: - Lines 57-70: The main idea should also be presented in one sentence. This section needs to be revised, as the main topic is trichinellosis in American mink in Lithuania. It is necessary to analyze and present data on whether mink infections with trichinellosis have been recorded in Europe or other countries, and on which continents.

Response 4: Thank you. We have corrected as suggested.

Point 5: Figure 1. Please indicate in the text or figure caption the program used to create the figure and its source.

Response 5: Thank you. We have added the program.

Point 6: "Reactions were carried out in a final volume of 25 µL, containing 12.5 µL of DreamTaq PCR Master Mix (Thermo Fisher Scientific, Vilnius, Lithuania), 0.5 µM of each primer, and 4 µL of template DNA." The total volume is 17.5 µL. What was the remaining volume?!

Response 6: Thank you, we corrected the sentence.

Point 7: Please transfer the sequenced and deposited sequence data (GenBank accession numbers), as well as the reference strain numbers used, to Section 2.4. DNA Sequence Analysis.

Response 7: Thank you. We have changed.

Point 8: Figure 2. The figure title needs to be changed; digestion was not artificial, but rather using artificial gastric juice.

Response 8: Thank you. We corrected the title of the figure.

Point 9: Line 198. "Ten Trichinella larvae were isolated from the infected animals." You're talking about multiple infected animals here, although line 191 reports one infected animal - "Ten Trichinella larvae were isolated from the infected animals...".

Response 9:  Thank you. We corrected from “infected animals” to “infected animal”.

Point 10: The authors report studying 10 larvae, but Figure 2 only shows 5 samples. Explain this.

Response 10:  Thank you for this comment. Figure 2 has been revised to include all 10 analyzed larvae. In the previous version, only 5 samples were shown for illustrative purposes of the electrophoresis gel results. The figure has now been updated for clarity and consistency with the text.

Point 11: Figure 2. N – negative control. What was used as the negative control? Include this in the figure title or in the description of the PCR method.

Response 11:  Added description to methods.

Point 12: Lines 220-251. There's too much descriptive text for the Results section; shorten it and move the description to the Discussion section.

Response 12: We have shortened the description of genetic results as suggested.

Point 13: The Discussion section, in my opinion, is also more descriptive and similar to the Introduction. The authors have divided the Discussion section into subsections. However, this section should contain comparative data obtained by you and other authors. Compare the numerical data, rather than simply listing where and in which countries trichinosis in minks has been detected.

Response 13: Thank you for your comment. We have significantly changed 4.1. part of the discussion as the reviewer suggested.

 

 

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

The authors have addressed all reviewer comments, and the manuscript has been substantially revised and improved. Although one issue appears to remain unchanged, the authors have stated that revisions were made; this matter ultimately remains at the authors’ discretion.

Point 7: Please transfer the sequenced and deposited sequence data (GenBank accession numbers), as well as the reference strain numbers used, to Section 2.4. DNA Sequence Analysis.

Response 7: Thank you. We have changed.