PONE-D-21-40040Synergistic effect of probiotic, chicory root powder and coriander seed powder on growth performance, antioxidant activity and gut health of broiler chickenPLOS ONE

Dear Dr. Gurram,

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Kind regards,

Kumar Venkitanarayanan, DVM, Ph.D.

Academic Editor

PLOS ONE

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- https://www.tandfonline.com/doi/abs/10.1080/00071660902806962?journalCode=cbps20

- https://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0260923

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Reviewer #1: Partly

Reviewer #2: No

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: No

Reviewer #2: No

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: No

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Summary: Interesting work. Please carefully review my comments and prepare a point-by-point response for the same. Revise the manuscript where appropriate. Thanks.

TITLE: Replace chicken with chickens.

Abstract:

Line 32: typo, please correct. Remove the extra “to”

Line 33: Please represent probiotic supplementation as percentage similar to chicory powder.

Line 37: IF you are representing antibiotic addition as gm/ton, please represent probiotic the same way. To avoid this confusion, I recommend representing your supplementations as percentage for all treatments.

Line 37: Please double check the abbreviation of gram. I believe it is “g”.

Introduction:

LINE 83: Remove the addition sign and replace with and.

Material and methods:

Line 108-113: Please include a table to describe the treatments clearly.

Line 116: Are the total bacterial load 32 billion CFU/100 g or each probiotic was at this concentration? Is this a commercial product or designed by the authors? How did the authors check the viability of each bacteria in the probiotic cocktail?

Line 120-122: Why are the authors presenting results in this section? I would recommend to move all the results presentation in the result section.

Line 118: Please represent probiotic levels as percentage too.

Line 130: “Feed intake”, please change to lower case F.

Line 134: Please include more details on how enzyme activity was determined. Describe how samples were stored, processed, followed by what type of assay was used.

Line 138: Please explain clearly how the 240 birds were sampled. It appears that only 1 bird/replicate that is 5 birds/treatment were processed for pH, intestinal contents etc. What about the other birds in the replicates? Why could the authors not process all birds for all samples to increase statistical power.

Question: How many trials/studies were conducted with 240 birds (6 dietary treatments, 8 replicates)?

Question: Where have the authors described the methodology for nutrient utilization? The results for nutrient utilization have been described in table 4 but I could not find the methodology portion for this section.

Question: How did the authors selectively enrich for Salmonella from the samples? What enrichment procedure was employed to facilitate recovery of Salmonella? Did the authors conduct a Salmonella load test at the start of the trials before the treatments began?

Results and Discussion:

Question: Why did the author not run groups for probiotic, chicory and coriander alone in the trials. Without these treatments, it is difficult to discern which component (probiotic, chicory, coriander) is effective in modulating body weight gains, FCR etc.

Table 7: Please share how the statistical analysis for comparing log CFU counts was conducted. I am interested to review log CFU/g values along with SE for each treatment. Some of the values for Lactobacillus (T5 7.88 vs T6 7.94), Salmonella (T4 7.33 vs T5 7.42) are very close to be statistically different from each other.

Reviewer #2: The authors investigated the effect of incorporating a combination of probiotics, chicory root powder and coriander seed powder on broiler growth performance, gut health and antioxidant activity. This study is relevant to the poultry industry for utilizing alternatives to antibiotic growth promoters.

However, my review for the manuscript was done partially because I felt that the experimental design and the number of birds used for the respective time points lacked clarity. It was hard to make an assessment on the data sets provided in the study. In addition, description provided for the statistical analysis and the inferences derived seemed incomplete. I would be happy to provide my further review and comments on the manuscript after the authors have provided a clarity on the experimental design and the N value for respective end points used in the study.

I have provided some of my comments for the manuscript below.

Abstract

Line 32-33 – please rephrase sentence as appropriate indicating the aim/background of the study. The details of the experimental design can be incorporated in the second sentence.

Introduction

Line76-77 – the connotation for antimicrobial properties of chicory root powder cannot be directly attributed but rather should be indicated as an indirect effect. Kindly rephrase the sentence appropriately. Or rather, if the authors had planned to indicate the antimicrobial activity other potential constituents, that may also be explained here.

Line 84 – the information regarding postbiotics may be removed from here. The authors can alternately provide additional references that cite the effect of probiotics + inulin combinations on poultry gut health. Specifically, the authors may cite the strains of probiotics that have been used by other researchers in a concise manner.

Line 90-92 – It would be better if the compounds listed in brackets are briefly expanded and described in terms of their chemical and functional properties/antimicrobial activities.

Materials and Methods

Line 116 – Does this mean the concentration of each bacterial strain or is this the total lactic acid bacterial counts in general?

Line 116 and Table 1 – Isn’t this analysis termed Proximate Analysis?

Line 118-121 – The way in which the treatment groups used for selecting the ideal formulation was not clear. What were the treatment groups involved in the broiler trials to determine the ideal composition of the mix that was eventually used for the final study?

Line 132 – clearly specify the N value for the metabolic trial.

Line 155-170 – how were the Salmonella colonies confirmed just by observed the colony morphology from the SS agar. What if the colonies observed in the plates were from Proteus?

Line 171 – was a blinded histopathological evaluation and analysis performed for the respective tissue samples?

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Reviewer #1: No

Reviewer #2: No

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PONE-D-21-40040R1Synergistic effect of probiotic, chicory root powder and coriander seed powder on growth performance, antioxidant activity and gut health of broiler chickenPLOS ONE

Dear Dr. Gurram,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Please submit your revised manuscript by May 16. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Kumar Venkitanarayanan, DVM, Ph.D.

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

Additional Editor Comments (if provided):

The following changes need to be done.

Lines 48-51 : Revise the sentence. Significantly appears twice in the sentence.

Lines 75-77: Recently, herbal feed additives products like chicory root powder are gaining attention as they indirectly promote antimicrobial 77 action by reducing the harmful bacteria in the gut. Replace “feed additives” with “feed additive”.

Confirm on the manuscript that all the treatment concentrations in the feed were weight/weight basis. If not, please indicate.

Lines 125-127: The dose levels (Probiotic at 10g/100 kg, chicory at 1.0 % and coriander at 1.5 % levels ) were selected based on my previous trail results. Delete “my”.

Line 368: Significant increase in the counts of Lactobacilli in test diets was also support the above results. Delete “was”.

Lines 503-511: Suggest revising the following text as below.

Supplementation of probiotic + chicory, probiotic + coriander, chicory + coriander and probiotic + chicory + coriander combinations produced greater weight gain, improved FCR, and higher antioxidant activity compared to control and antibiotic at 42 d of age. The combination of probiotic (0.01%) with chicory root powder (1.0%) was more effective than combinations of other additives in terms of body weight gain and FCR. Supplementation of different combinations of probiotic, chicory root powder and coriander seed powder significantly lowered the gut pH, E. coli and Salmonella counts and increased the Lactobacilli counts. In addition, this treatment improved the gut morphometry parameters such as VH, CD and VW in the in the intestines. Thus, supplementation of probiotic at 0.01%, chicory root powder at 1.0 %, and coriander seed powder at 1.5 % combinations could be used in the diet as a potential growth promoter in broiler chickens. However, follow up large-scale studies under field conditions are necessary before recommending the compounds in the broiler diet.

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[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

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Synergistic effect of probiotic, chicory root powder and coriander seed powder on growth performance, antioxidant activity and gut health of broiler chickens

PONE-D-21-40040R2

Dear Dr. Gurram,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

Kumar Venkitanarayanan, DVM, Ph.D.

Academic Editor

PLOS ONE