Consumption of Goat Cheese Naturally Rich in Omega-3 and Conjugated Linoleic Acid Improves the Cardiovascular and Inflammatory Biomarkers of Overweight and Obese Subjects: A Randomized Controlled Trial

Round 1

Reviewer 1 Report

The authors examined the value of a goat cheese naturally enriched in PUFA n-3 and CLA as means of improving cardiovascular and inflammatory health of overweight and obese subjects. Consumption of this cheese caused significantly changes in plasma total- cholesterol and HDL-cholesterol, as well as in apolipoprotein B and high-sensitivity C-reactive protein concentrations. The authors suggest that the consumption of a PUFA enriched cheese can obtain significant improvement of the plasma lipid profile and inflammatory status and proposes a potential role of this dairy product as an alternative attractive to develop high nutritional value food in a balanced diet comprising regular exercise.

The authors no report any table on plasma fatty acid profile in subjects supplemented. It would be interesting and would make the manuscript more complete.

The authors can verify the number of tables? It’s not present table 2 and in manuscript the authors report table 2 to indicate the "baseline characteristics" that it is indicated with table 1.

In line 76 the authors can correct “oildseeds”

In table 1 the authors reported total fat mass (%) around 46 and lean mass (%) are the same. I dont understand. This % are correct? The total fat mass include android fat and gynoid fat? the authors can verify the values?

In table 4 the authors can indicate with "*" where values is significant, because in line 228 the authors report a "significant" improvement in the LDL/HDL ratio.

In line 245 to correct the symbol in TNF

In line 260 to correct  “hipocaloric”

In line 293 to correct “hipercholesterolemic”

In line 298-301 the authors write that “Carrero et al. (2007) [ref. 29] supplemented hyperlipidemic volunteers with a milk product containing EPA plus DHA and observed a significant reduction in TAG and TC after 8 week, but probably this effect was due to oleic acid, folic acid, and vitamin B6 added to the milk”. This significant reduction in TAG and TC maybe can dependent by EPA and DHA not oleic acid, as other authors have seen (Pintus 2013 and Murru 2018). Moreover in Santurino 2017 no change the oleic acid concentration in control and enriched cheeses. The authors can verify this information and can complete the discussion?

In line 348-356 the authors report same sentences of conclusions.

 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

This paper presents the effect of n-3 PUFA and CLA-rich goat cheese compared to control goat cheese on variuous markers of cardiovascular risk and nutritional status, in the context of a hypocaloric dietary intervention in overweight and obese subjects a high cardiovascular risk. The amount of data collected is impressive, but appears somewhat random. Many details are missing in the method, and the presentation of the results and statistical analysis needs to be clearer. The general flow of the manuscript could be improved by justifying the outcomes measured, and making method, results and discussion more consistent. The language could also be improved to be clearer and simpler, which would facilitate reading.

More specific (but not exhaustive) comments are listed below to help improve the manuscript.

 

Abstract

L31-33: It would better to be more specific: significantly increased / decreased... compared to control goat cheese

 

 

Introduction

L49-50: Avoid using 'and' twice. break down the snetence if needed

L52: remove 'of healthy habits'

L53: it is unclear whether the authors are talking about dietary cholesterol or lipoprotein rpofiles

L54: 'are key to the prevention...'

L56: pls make sure the Greek symbol alpha shows correctly, write n-3 rather than n3 (and n-6 rather than n6 all throughout the manuscript)

L59: ref number is wrong. Perhaps referring to meta-analysis would be more convincing when referring to the effect of n-3 PUFA on lipoprotein profiles.

L60-62: the sentence is missing a reference

L63-64: reference needed

L65: The reference provided refers to n-3 PUFA rather than CLA

L72-73: reference to meta-analysis is missing

 

Material and methods

L90: the recruitment of participants (where, when and how) need to be described

L95: Rephrase to ‘Inclusion criteria were : …’

L101: Were people with lactose intolerance also excluded?

L103: the hypocaloric diet should be described in more details, how many cal per day or per kg body weight? The nutritional composition of the cheeses (control and enriched) also needs to be described (particularly the PUFA and CLA content in both cheeses).

L112: remove ‘placebo’ and ‘group and’

L113-114: ‘whose … loss’ should be simplified: ‘previously shown to induce significant weight loss’

L119-120: Thus should replace ‘so that’, and ‘know’ should be replaced by ‘knew’

L121: not needed

L123: not sure what dietetic data is, do you mean dietary intake?

L124: why were both FFQ and 3 day food diary used?

L128-132: it is unclear what was performed only at baseline and every 3 weeks. MET stands for Metabolic Equivalent of Task

L134: the conditions in which BP and HR were measured must be described: seated / supine position? after how long rest? how many times?

L141-152: specific which vacutainers was used for blood collection (EDTA, etc). More details need to be described: centrifugation of blood, incubation times, etc? or refer to previous literature if available. Ideally there should be a separate part on ‘biochemical analysis’. Some outcome measurements are also missing: apoA-1 and apoB, fasting glucose, fasting insulin, HbA1c, OxLDL and fibrinogen, calcium, phosphate, vitamin D, ghrelin and leptin, mentioned later in the statistical analysis. The measurements of all these outcomes should be justified.

L160: have consent forms also been obtained at the time of enrolment?

L173: what are ‘range of change’? The statistical analysis is unclear. in particular, how were the changes from baseline compared between the groups? t-tests? The analysis should also mention when data when not normally distributed, and if transformation of the data or a non parametric test was required.

 

Results

L194: remove * in the legend (there is no significant difference that appears in the table), specify the test used to provide the p values. Baseline characteristics do not include values at 12 weeks, but changes from baseline should be described in another table, with statistical analysis of the changes.

L208: it would be good to specify if BP changed from baseline, (in addition to between the groups), it may have significantly increased / decreased even if it remained in the normal range

L212-220: It would be good to point out that changes in TC were significantly increased in EG compared to CG, despite no significant difference from baseline observed in either group. In general the presentation of the results needs to be simpler and clearer: it is difficult to tell whether the authors are referring to changes form baseline and differences at 12 weeks.

L221-25, 241-46: Tables 3 and 5 could be simplified, the right hand side of the table is unclear. What are the values in brackets in the CG and EG columns? Is IC95% actually the difference between EC and EG as mean (95%CI)? The table should be understood by itself, with its legend and no need to refer to text. Specify in the legend of Table 3 if t-test / Mann-Whitney test were used to compare means between groups. It is unclear if # shows differences in values at 12 weeks or differences in changes from baseline.

L226-236: It is unclear if both LDL/HDL ratio and TC/HDL-C ratio are changed or just the former. Significant differences (with test used) should also appear in Table 4. It would be better to present all tables in the same way to facilitate reading.

Certain outcomes mentioned in the statistical analysis are missing (e.g. HOMA, HbA1c). There is also no report of dietary intake measured by FFQ or food diary.

 

Discussion

All throughout, replace hipo / hiper with hypo / hyper

L264: lipid profiles

L267: it was unclear that physical activity was part of the intervention, this should be explained in the methods. Results should include how much the participants complied to physical activity guidelines, perhaps using MET as described in the method?

L268: there is confusion between waist circumference and metabolic risk

L270: use heart rate rather than frequency.

L272: CVR needs to be explained

L277: Table 2 doesn’t show the statistical analysis of the changes from baseline, which would allow to say whether there was a decrease in TFM etc in both groups

L278-81: What is BMD and how does it fit into the paper? It does not appear in intro, method or results until then. Results on physical activity are missing

L299: The effect of n-3 PUFA on triglycerides is supported by several robust meta-analyses

L308-309: The results did not show whether there was a significant difference in ApoB/ApoA1 ratio

L318: What is FFA and how does it fit into the paper? It does not appear in intro, method or results until then.

L331-334: the relationship between the incorporation into cell membranes and the anti-inflammatory effect of n-3 PUFA is not clear.

L335-347: The relevance of the measurement of calcium, phosphore, ghrelin or leptin in the context of this study is not clear

L350: What is CCi and how does it fit into the paper? It does not appear in intro, method or results until then.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

This new version of manuscript can be ok. The authors have modified the  discussion satisfactorily. Thank you

Author Response

Please see the attachment

Author Response File: Author Response.pdf