The Possibility of Using the Probiotic Starter Culture Lacticaseibacillus rhamnosus LOCK900 in Dry Fermented Pork Loins and Sausages Produced Under Industrial Conditions

Round 1

Reviewer 1 Report

The manuscript titled “The possibility of using the probiotic starter culture Lactobacillus rhamnosus LOCK900 in dry fermented  pork loins and sausages produced under industrial conditions” investigates the possibility of transferring laboratory based meat processing technology to the commercial settings. The article is interesting, however, I can’t see the additional value of the work done. Therefore I recommend Major Revision. Other specific comments are as follows:

1. The article needs English proof reading e.g. line 28, line 43 (published not publish), line 49 (these not this), line 74, line 260, line 330 (loins not lions).
2. Line 48: It is enough to say meat.
3. Lines 57-59: What is the additional value of this research? What were the shortcomings of other research presented in the literature? What these studies concerned? Please provide citations.
4. Lines 65-66: Please highlight the additional value of the research by naming technology transfer constraints.
5. Table 2: Please double check the indication of statistically significant differences e.g. looking at the mean and SD there shouldn’t be significant difference between control sausages and probiotic loins. I would also advise you to reorganize tables so that loins and sausages are separately compared.
6. Discussion: Please divide this section similarly to the Results section.
7. I believe you should more focus on the differences concerning two productions methods: laboratory and industrial. At the moment as they are presented the results just confirm what is known.

Author Response

Response to Reviewer 1 Comments

Thank you for any comments and suggestions. The amendments have been marked in red in the text. Below are the answers to the Reviewer's 1 comments:

Point 1:The article needs English proof reading e.g. line 28, line 43 (published not publish), line 49 (these not this), line 74, line 260, line 330 (loins not lions).

Response 1: The article has been checked and corrected by a native speaker.

Point 2:Line 48: It is enough to say meat.

Response 2:Done.

Point 3: Lines 57-59: What is the additional value of this research? What were the shortcomings of other research presented in the literature? What these studies concerned? Please provide citations.

Point 4: Lines 65-66: Please highlight the additional value of the research by naming technology transfer constraints.

Response 3 and 4: Using of starter cultures in the production of dry fermented meats is always intentional and aims at obtaining the specified sensory and microbiological characteristics of the end product. Currently the production of commercial starter preparations use primarily lactic acid bacteria which show favourable technological effect. Therefore, the additional value of presented research is primarily the use of probiotics as starter cultures in dry fermented meat production. An added value is also the fact that an effective technology for producing dry fermented meat products under industrial conditions has been developed, which technology is not commonly practiced in meat processing in the world. There are many reasons. First, the selection of appropriate probiotics for sausage production is difficult due to their sensitivity to technological conditions. As microorganisms isolated from the human digestive tract, these must dominate the non-sterile environment of the meat raw material. The second reason is the development of such  technology that will allow the development of probiotics in non-minced meats, e.g.loin. The authors raised this problem in discussing of the results. The main limitation is the lack of clinical studies of developed dry fermented meat products,  confirming the survival of used probiotic strain in the human digestive tract. This element is a further goal of the authors' research.

Point 5: Table 2: Please double check the indication of statistically significant differences e.g. looking at the mean and SD there shouldn’t be significant difference between control sausages and probiotic loins. I would also advise you to reorganize tables so that loins and sausages are separately compared.

Response 5: Done. Thank You.

Point 6: Discussion: Please divide this section similarly to the Results section.

Response 6: The proposed discussion section was organised in accordance with the order of the presented research results. In the authors' opinion, this method of presentation allows better comparison of individual batches of performed tests with each other. It also creates a holistic picture of final products: potential probiotic dry fermented sausages and loins.

Point 7: I believe you should more focus on the differences concerning two productions methods: laboratory and industrial. At the moment as they are presented the results just confirm what is known.

Response 7: The authors' proposal is in the text: Line 443-449.

Author Response File: Author Response.pdf

Reviewer 2 Report

The manuscript entitled "The possibility of using the probiotic starter culture Lactobacillus rhamnosus LOCK900 in dry fermented pork loins and sausages produced under industrial conditions" provides valuable information and may be published in "Applied Sciences"   I have only minor remarks.   Line 18. Please use fuff name of LAB. It's first appearance of this abbreviation in the text
Line 26. I have the same remark concerning LOCK900
Lines 42-44. Is this sentence really necessary?
Line 77. References cited there should be cited also in subsections 2.1.1 and 2.1.2. Please add also complete list of abbreviations.

Author Response

Response to Reviewer 2 Comments

Thank you for any comments and suggestions. The amendments have been marked in red in the text. Below are the answers to the Reviewer's 2 comments:

Point 1: Line 18. Please use full name of LAB. It's first appearance of this abbreviation in the text.

Response 1: Done.

Point 2: Line 26. I have the same remark concerning LOCK900

Response 2: The abbreviation “LOCK900” was used for the first time in the title. Changes to the abstract will not be in line with the magazine's requirements (max. 200 word).

Point 3: Lines 42-44. Is this sentence really necessary?

Response 3: The sentence and whole paragraph were reorganized. Please see line 42-43.

Point 4:Line 77. References cited there should be cited also in subsections 2.1.1 and 2.1.2. Please add also complete list of abbreviations.

Response 4: Deficiencies have been corrected and supplemented.Thank You.

Author Response File: Author Response.pdf

Reviewer 3 Report

The title of table 1 is not clear. Please make it more comprehensible. 

Please describe the curing mixture (line 100).

Cold smoke was produced using which wood or sawdust...?

 

 

Author Response

Response to Reviewer 3 Comments

Thank you for any comments and suggestions. The amendments have been marked in red in the text. Below are the answers to the Reviewer's 3 comments:

Point 1: The title of table 1 is not clear. Please make it more comprehensible.

Response 1: The title has been changed to “Dry fermented pork loins and sausages samples and their raw material composition (addition per kg of meat)”.

Point 2: Please describe the curing mixture (line 100).

Response 2: The pork loins were then covered in curing mixture (99.5% sea salt, 0.5% sodium nitrite) at a ratio of 2.8% to the meat content.

Point 3: Cold smoke was produced using which wood or sawdust...?

Response 3: A cold smoke was produced using alder wood.

Reviewer 4 Report

The research paper applsci-825793- entitled: The possibility of using the probiotic starter culture Lactobacillus rhamnosus LOCK900 in dry fermented pork loins and sausages produced under industrial conditions is interesting. It fits with the scope of the journal. The author developed technology for the production of dry fermented meat products supplemented with probiotics, and successfully transferred to industrial conditions. The authors used both culture-dependent and independent techniques to asses the viability and identify the probiotic strain in the product.

Two minor queries need to be addressed before publication

 

Overall:

The genus Lactobacillus has been reclassified, L. rhamnosus is named differently according to a new publication, the authors should check and correct the bacterial names in the text accordingly

A web-based tool has been developed to help to determine the new names of all Lactobacillus species http://lactotax.embl.de/wuyts/lactotax/

Specific comments:

Introduction

line 40: please provide the correct definition of probiotic, according to the WHO (2001), and updated by the panel of experts from ISAAP (Hill et al, 2014): https://www.nature.com/articles/nrgastro.2014.66:

Author Response

Response to Reviewer 4 Comments

Thank you for any comments and suggestions. The amendments have been marked in red in the text. Below are the answers to the Reviewer's 4 comments:

Point 1 (Overall): The genus Lactobacillus has been reclassified, L. rhamnosus is named differently according to a new publication, the authors should check and correct the bacterial names in the text accordingly

A web-based tool has been developed to help to determine the new names of all Lactobacillus species http://lactotax.embl.de/wuyts/lactotax/

Response 1: The correct, new name of Lactobacillus was used. Changed to: Lacticaseibacillus; Lacticaseibacillus rhamnosus

Point 2 (Specific comments): Introduction. line 40: please provide the correct definition of probiotic, according to the WHO (2001), and updated by the panel of experts from ISAAP (Hill et al, 2014): https://www.nature.com/articles/nrgastro.2014.66:

Response 2: The correct definition of probiotic was added.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

The authors have improved the manuscript. However, I believe it still needs to be revised. 

In my opinion the authors still haven't justifies the research properly. They state in line 58 that there are limited reports on practical aspects of implementing production using probiotic bacteria. I think they should cite this references and on this background present the additional value of their research. 

Additionally in lines 443-449 there is no proposal regarding remark no 7. The authors did not mark the new text in red either.

Author Response

Response to Reviewer 1 Comments (Round 2)

Thank you for reviewing our manuscript again. All changes made in accordance with the comments were marked using the function in Microsoft Word "Track Changes".

The answers to the Reviewer's 1 comments:

Point 1: The authors have improved the manuscript. However, I believe it still needs to be revised.

In my opinion the authors still haven't justifies the research properly. They state in line 58 that there are limited reports on practical aspects of implementing production using probiotic bacteria. I think they should cite this references and on this background present the additional value of their research.

Response 1: Thank you for paying attention to the inaccurate wording regarding limited reports on practical aspects of implementing production using probiotic bacteria. The changes made by the authors are in lines: 52-55, 61-66, 70-78.

Point 2: Additionally in lines 443-449 there is no proposal regarding remark no 7. The authors did not mark the new text in red either.

Response 2: Lines 443-449 were concerned subsection "Author contributions". There was a mistake in the designation of the lines and no marked text in red. I apologize for inaccuracy and error. The changes introduced by the authors are in line 430-436 and concern:

“Laboratory and semi-technical studies regarding the design of new meat products have been previously carried out under standardized conditions [16,17,20,21,22]. This applies to both strictly observed hygiene conditions, as well as the quantity and weighing of individual raw material additives. Particular attention should be paid to addition of a probiotic starter culture. In laboratory conditions the probiotic strain was added to the meat matrix under sterility conditions (the equipment and qualifications of the people), which was not impossible in the production hall of the industrial plant. However, it was found that the developed technology for the production of two kinds of dry fermented meat products (pork loins – non-minced meat and sausages – minced meat) with the addition of the probiotic strain LOCK900 has been successfully transferred to industrial conditions. Based on the results of this study and by other authors' it can be stated that both the hygiene criterion of the production process and the criterion of food safety have been met. Moreover, the added value of manufactured meat products is in the use of the probiotic starter culture LOCK900 with pro-health and bio-conserving properties”.

The above text has already been clearly highlighted and concerns the introduction / explanation of the specifics in the production of potential probiotic dry fermented meat under laboratory and industrial conditions.

 

Author Response File: Author Response.pdf