Protozoa as the “Underdogs” for Microbiological Quality Evaluation of Fresh Vegetables
, Susana Sousa 1,2,3, António Castro 1,2,3, Vânia Ferreira 4, Paula Teixeira 4,* and José M. Correia da Costa 1,2,3
Round 1
Reviewer 1 Report
Thank you very much for the oportunity to read sucha great work. The problem of protozoa contamination may be very important in for protecting the society from food-delivered infecions. However most of published studies focus on the microbial or fungi problem. Therefore, this article has a significant level of novelty.
The introduction is written very well and introduce into the problem of protoza contamination. M&M section provide sufficient information about metodology and also show the experience of Authors in this area. Results are desribed clearly and discussed.
In my opinion the article could be even better with Conclusion section containg information about possibilities of protozoa detection in various stage of food production.
The authors should also check the manuscript and delete "doubled-spaces" (for example in line 24, 56, 215).
Author Response
Reviewer 1
Thank you very much for the oportunity to read sucha great work. The problem of protozoa contamination may be very important in for protecting the society from food-delivered infecions. However most of published studies focus on the microbial or fungi problem. Therefore, this article has a significant level of novelty.
The introduction is written very well and introduce into the problem of protoza contamination. M&M section provide sufficient information about metodology and also show the experience of Authors in this area. Results are desribed clearly and discussed.
In my opinion the article could be even better with Conclusion section containg information about possibilities of protozoa detection in various stage of food production.
Thank you very much for your comments and opinion.
A conclusion section containing the suggested information was added to the manuscript.
The authors should also check the manuscript and delete "doubled-spaces" (for example in line 24, 56, 215).
As suggested, all "doubled-spaces" have been checked and deleted.
Reviewer 2 Report
This article has been well written. However, there is still room for improvement.
Results and Discussion
11. Please include the epifluorescence image for the samples with positive findings.
22. Although you have done the bacteria analysis, no result has been included in the manuscript. Please do so.
33. Do you think 7 out of 16 samples can justify the importance of protozoan analysis in RTE vegetables? Please include it in the discussion section.
44. Why do you think salad is more prone to the high prevalence of protozoan parasites than other vegetables? Please include it in the discussion section.
Other comments:
55. Do you have any water sample results to support contamination claimed after harvesting the crops?
Author Response
Reviewer 2
Dear Authors,
Taken into account the experience and learning obtained from the SafeConsume work package 2 (WP2) laboratory studies, and the “One Health” approach, a parallel preliminary work was performed in order to access the microbiological quality of fresh vegetables highlighting not only bacteria (E. coli, L. monocytogenes and Salmonella spp.) covered by the legislation, but also the zoonotic protozoa G. duodenalis and Cryptosporidium spp.
Undoubtedly, this short communication is very gripping as well as it has a scientific value.
The introduction provides a good, generalized background of the topic that quickly gives the reader an appreciation of the scientific relevance and timeliness of the research theme. I think the motivations for this study are very clear. It is also worth noting the extremely detailed description of the section Materials and methods.
The experimental apparatus is appropriate for the study. The tables are easy to interpret.
The obtained findings are properly described in the context of the published literature. Also the conclusions of the study were supported by appropriate evidence.
Thank you very much for your comments and opinion.
I have for Authors just minor suggestions for improvement as follows:
Line 213: UFC/g – what is the unit?
UFC has been replaced by the English abbreviation of Colony-forming unit, CFU.
Table 2: Please, replace commas with dots.
Table 2 has been edit as suggested by the reviewer.
Please, complete in the section References, the cited in section Materials and methods - ISO standards.
As suggested, ISO standards have been included in the reference section.
From my standpoint, the presented manuscript is appropriate for publication in Journal Applied Sciences, after minor revision, given the above aspects.
Reviewer 3 Report
Dear Authors,
Taken into account the experience and learning obtained from the SafeConsume work package 2 (WP2) laboratory studies, and the “One Health” approach, a parallel preliminary work was performed in order to access the microbiological quality of fresh vegetables highlighting not only bacteria (E. coli, L. monocytogenes and Salmonella spp.) covered by the legislation, but also the zoonotic protozoa G. duodenalis and Cryptosporidium spp.
Undoubtedly, this short communication is very gripping as well as it has a scientific value.
The introduction provides a good, generalized background of the topic that quickly gives the reader an appreciation of the scientific relevance and timeliness of the research theme. I think the motivations for this study are very clear. It is also worth noting the extremely detailed description of the section Materials and methods.
The experimental apparatus is appropriate for the study. The tables are easy to interpret.
The obtained findings are properly described in the context of the published literature. Also the conclusions of the study were supported by appropriate evidence.
I have for Authors just minor suggestions for improvement as follows:
- Line 213: UFC/g – what is the unit?
- Table 2: Please, replace commas with dots.
- Please, complete in the section References, the cited in section Materials and methods - ISO standards.
From my standpoint, the presented manuscript is appropriate for publication in Journal Applied Sciences, after minor revision, given the above aspects.
Author Response
Reviewer 3
This article has been well written. However, there is still room for improvement.
Thank you very much for your comments and opinion.
Results and Discussion
- Please include the epifluorescence image for the samples with positive findings.
As suggested, representative epifluorescence images of Cryptosporidium and Giardia positive samples were added to the manuscript.
- Although you have done the bacteria analysis, no result has been included in the manuscript. Please do so.
Results are presented in table 2 for L. monocytogenes and in supplementary table for all the other parameters.
- Do you think 7 out of 16 samples can justify the importance of protozoan analysis in RTE vegetables? Please include it in the discussion section.
Based in our results and other previous studies, it is clear that current RTE production processes do not guarantee a product free from parasites (Caradonna et al. 2017, Marques et al 2020). Therefore, it is essential to adopt the actions stated in the last paragraph of the manuscript (including rapid and reliable parasite detection), in order to minimize the presence of protozoa in the different stages of food production, including agricultural production and processing. This idea was reinforced in the discussion section, as suggested.
- Why do you think salad is more prone to the high prevalence of protozoan parasites than other vegetables? Please include it in the discussion section.
Eating raw, or slightly cooked vegetables is one of the means by which the transmission of these parasitic infections are disseminated. Changes in eating habits, to healthier diets, have resulted in increased consumption of salads, exposing consumers to these parasites; and that is why this type of fresh product is more prone to be contaminated. This justification has been included in the conclusion section.
Other comments:
- Do you have any water sample results to support contamination claimed after harvesting the crops?
Unfortunately, we did not collect samples from irrigation water and water used for washing RTE vegetables. However, as water has been identified as a relevant risk factor for foodborne pathogens, this is a very important subject that currently we are trying to address.
Reviewer 4 Report
Important subject and interesting results.
The title is a bit inappropriate for scientific journals.
The small number of samples (only 16).
This research is based on previous work.
Author Response
Reviewer 4
Important subject and interesting results.
Thank you very much for your comments and opinion.
The title is a bit inappropriate for scientific journals.
We appreciate the reviewer opinion, but we still think that the title reflects and highlights its content and importance. However, if the reviewer or the editor suggest a more suitable title, we promptly will accept it.
The small number of samples (only 16).
Indeed, we agree with the reviewer that the number of samples is quite low. However, this is a preliminary study written as a short communication. Moreover, the publication of this study does not exclude further studies to increase the number of samples, but also, using other type of vegetables, type of agriculture, collection season, irrigation water analysis etc.
This research is based on previous work.
Yes, the research is based on previous work, but results were not published before.
Round 2
Reviewer 4 Report
Thank you for the comments.
