Assessing the Microbiological Safety Parameters of Minas Artisanal Cheese Samples in Retail Environments in São Paulo, Brazil

Round 1

Reviewer 1 Report

The paper entitled "ASSESSING THE MICROBIOLOGICAL SAFETY PARAMETERS OF MINAS ARTISANAL CHEESE SAMPLES AT RETAIL IN SAO PAULO, BRAZIL" is an interesting and well-written work. However, conclusions have to be improved and extended.

Author Response

The authors thank Reviewer 1 for the positive comments. Several parts of the manuscript were reviewed, including the conclusion session, as suggested. 

Author Response File: Author Response.docx

Reviewer 2 Report

General comment

The present work aimed to evaluate microbiological safety parameters of Minas artisanal cheeses collected at retail in Sao Paulo (Brazil) in order to assess the exposure of consumers to common pathogens. However, several studies showing that artisanal chesses produced in Brazil still do not satisfactorily meet the microbiological criteria established by the national laws have been just published (e.g. Camargo, A.C., et al. 2021). Hence, this present study was not original and no useful conclusions are provided by authors. Moreover, this study showed several drawbacks as specified in the following comments and it could be edited by a native speaker to improve some phrases from a grammatically point of view. For all of these reasons, I think that this manuscript cannot be accepted for publication.

Specific comments

Line 36: no comma at the end

Line 37-38: authors should add more information about the particular characteristics of this cheese. Here they are just mentioned but additional  it could be very interesting for the reader

Line 40: the correct abbreviation of the Institute of National Historical & Cultural Heritage is ISHAN and not IPHAN as reported by authors.

Line 50-52: “Regulatory agencies around the world have established quality and safety guidelines 50

and parameters for these products, to protect the traditional production procedures and 51

ensure products suitable for consumers, and Brazil is not an exception”. This comment is not useful for the reader.

Line 64-67: “Usually, retailers purchase the entire cheeses from the producers and cut them into smaller pieces for commercialization. It is a common practice that these cheese pieces are kept at room temperature, instead of refrigeration.” Why the entire cheeses need refrigeration while pieces can be stored at room temperature? What is the difference? Authors need to clarify this aspect because is not clear.

Line 86-88: “Each analytical unit consisted of two portions of 12.5 ± 0.1 g, one taken from the external part of the cheese and the other from the internal part of each sample.” Authors reported sampling of two units: one for external part and another for the internal part of cheese. This sampling form was mandatory by guidelines? This is not clear from the text. Otherwise, it could be better to consider two units from external and two units from internal part of the same product.

Line 103-105. Real-time PCR conditions were not reported by authors. Please, indicate cycle of amplification.

Table 1 was not necessary because primers used in this work were just previously published.

Line 201-207: this is an introductory part that could be removed from the discussion section.

Line 208-212: authors reported some factors that could be useful to reduce Salmonella contamination incidence, but it is not so sure that these factors are responsible for the low presence of pathogen in this study. Authors should perform additional analyses to demonstrate this aspect.

Line 213-216: Authors reported that Salmonella presence can be strongly influenced by E. coli and staphylococci contamination. Can this sentence be supported by references?

Line 225. Add a dot between [36] and Coliforms.

Line 301-305: please, use the same font throughout the text

 

Author Response

The authors thank Reviewer 2 for the comments. Despite not recommending the acceptance of the manuscript, the comments were interesting and constructive, and helped to improve the text. We hope that Reviewer 2 will change the evaluation after reading the new version of the manuscript.

Specific comments are listed below: 

• Typing mistakes in “old” Lines 36, 37-38, 40, 86-88, 225 and 301-305 were corrected.
• Line 64-67: the paragraph was rewritten, to address the issues raised by the reviewer.
• Line 103-105: the cycles of amplification were not described as they followed the instructions of the manufacturers, available in the package inserts. The authors prefer to keep this information as presented in the original text.
• Table 1 was maintained, as the authors believe it helps the reader.
• Lines 201-207: the sentence was removed.
• Line 208-212: the sentence was rephrased for better clarity. Unfortunately, it is not possible to perform additional analyses at this point.  
• Line 213-216: the sentence was rephrased for better clarity.
• The conclusion session was improved, and included the suggested reference of Camargo et al, 2021.

Author Response File: Author Response.docx

Reviewer 3 Report

The title of this study is: Assessing The Microbiological Safety Parameters Of Minas Artisanal Cheese Samples At Retail In Sao Paulo, Brazil. In this study was to provide data on the hygienic-sanitary conditions and microbiological safety of Minas artisanal cheese samples, collected at retail level in the city of São Paulo, through the investigation of the occurrence of Listeria monocytogenes and Salmonella spp, and counts of coagulase positive enterotoxigenic staphylococci, Escherichia coli and total coliforms, using both plating and molecular techniques.

I commented on the manuscript and the comments are presented below:

Part 1: Introduction.

The Introduction to the study is too broad and does not end with a clearly stated p urpose or goals that the Authors wish to pursue. This should be changed. I suggest supplementing the Chapter with additional information related to other new methods and devices in studied research. There are two characters at the end of a sentence on line 36: a period and a comma in red. The comma should be removed.

 

Part 2: Material and Methods

The Methods section provides the reader with enough information to repeat the experiments conducted. Were the cheeses received on delivery or only available on the shelves in stores? It is important due to the possibility of determining the initial parameters, i.e. the state before examining the protection conditions in the store and comparing the obtained results with each other in order to determine the protection. With such a large number of parameters tested that may affect the tested features, a more advanced statistical analysis should be performed. Only the basic statistical analysis was used to describe the differences. Have the Authors attempted to use other more comprehensive statistical analyzes. The use of advanced statistical methods is helpful for a full description of the relationship between the studied parameters and aspects of research conducted in the presented manuscript. You can determine the strength of the influence of a particular parameter on the variance of the system. At the same time, correlation relationships between the determined parameters can be determined.

Part: 3 Results and discussion

For the most part the Results section is well structured.

In the Discussion chapter, there is no full comparison and confrontation with the research of other authors in this area. The results were not fully discussed. A full discussion of the results obtained with other work in this field should be carried out in more aspects. I suggest supplementing the Chapter with additional information.

Part: 4 Conclusion

The Conclusions chapter contains information obtained after conducting experiments but performing only base statistical analyzes and were no comparison and confrontation with the research of other authors in this area.

Part: References.

The literature used is appropriate but should be supplementing about the items from the last years of publication about similar problem. The literature should be supplemented with additional items describing the examined aspects.

Author Response

The authors thank Reviewer 3 for the interesting comments. The text was thoroughly revised, and several paragraphs were rewritten.

The Introduction was reduced, the objectives were better explained and the typing errors were corrected.

The Materials and Methods part was modified, to address the questions raised by the reviewer. As for the missing statistical analyses, the authors believe they do not apply, as this study aimed to investigate the microbiological characteristics of Minas artisanal cheeses sold in the market of the city of São Paulo, simulating the purchase by a common consumer. The objective was to evaluate the consumer exposure to some pathogens and hygiene indicators, and not evaluate the influence or correlation of any parameter of the cheese chain with the observed results.

As for the Discussion and Conclusion chapters, there is a plethora of publications on microbiology of artisanal cheeses in Brazil and around the world, and a full discussion of all them would turn the reading too tiring. However, a new sentence was added to the Conclusion session, explaining that the study complements a recent systematic review published by Camargo et al, 2021, indicating that artisanal cheeses produced in Brazil still do not satisfactorily meet the microbiological criteria established by the regulators.

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

Authors improved the old version of this manuscript following all provided suggestions. In this version, the paper can be accepted for the pubblication. However, the sentence on line 234 "Most of the Minas artisanal cheese samples positive for Salmonella spp. (87.5%) were 233 also positive for total coliforms, E. coli and coagulase positive staphylococci (Table 1)" contains an error because Table 1 reports the sequences of primers not results. 

Author Response

We thank the reviewer for noticing this typing error. It was corrected (Table 2 instead of Table 1).

Reviewer 3 Report

The authors referred to the comments from the previous review for the manuscript titled: Assessing The Microbiological Safety Parameters Of Minas Artisanal Cheese Samples At Retail In Sao Paulo, Brazil. I accept explanations. In the future, I suggest using more precise  describing relationships between the parameters studied. They supplemented the discussion with a new literature data strengthens the message and importance of information in the manuscript. 

Author Response

The authors thank the reviewer for the positive evaluation of the revised version. The relationships between the parameters studied and the outcomes are subject of an ongoing research project.