Influence of Transport Distance, Animal Weight, and Muscle Position on the Quality Factors of Meat of Young Bulls during the Summer Months

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The work presented is on a topic that is still completely unresolved and still timely.

However, I have several reservations about the methodological part of the experiment.

It is necessary to specify which specific muscles were taken for the experiment. Labelling only the meat cuts is scientifically insufficient. The shoulder clod consists of multiple muscles and if it is not possible to declare that samples were consistently taken from the same location, the whole research is meaningless.

I believe that an analysis based on the calculation of correlation coefficients is not entirely appropriate. Firstly, the number of observations in some groups is insufficient, and secondly correlation does not imply causation. More appropriate statistical analysis options are available.

2.4 Sensory analysis:

The method of storing meat for sensory analysis is quite strange and quite easily allows inconsistent treatment of all samples equally.

It is necessary to indicate how the descriptors were assessed. It is necessary to indicate the scale; it is not obvious at first sight whether a higher value is more favourable or vice versa.

Can you indicate how many replicate samples overall and for each treatment were evaluated?

2.5 Shear force

This is a rather unusual combination in which the samples were evaluated. The WBSF is usually used to measure intact lean muscle mass. For minced meat, TPA is recommended. Explain why you have applied this procedure. It is then difficult to compare values with the cited literature, which takes a different approach. Describe better the dimensions of the cut samples.

2.6 Statistical analysis

The statistical model for sensory analysis should account for the judge effect.

There is no mention of how the correlations are calculated.

 

 Specific comments

L47: hardness !

L54: A significantly longer time interval than 7 to 21 days is recommended to achieve the favourable organoleptic characteristics of dry-aged meat. This is also stated in the literature cited.

L141: Justify why blooming time 0 was applied? Te is not the usual way of measuring the colour of beef.

L170: change WSBF to WBSF (to keep it consistent throughout the text)

L424: I don't think it is appropriate to refer to silverside as low value cuts. It may not be suitable for steak preparation, but the main muscle (BF) is often described as economically important or valuable. Please rephrase that sentence.

Author Response

Response to reviewer 1

The work presented is on a topic that is still completely unresolved and still timely.

Thank you very much for the constructive comments about our research. We were very pleased to have you as our reviewer and to discuss about your point of view about our work. We tried to follow your suggestions to enhance the quality of our paper.

Q1: However, I have several reservations about the methodological part of the experiment. It is necessary to specify which specific muscles were taken for the experiment. Labelling only the meat cuts is scientifically insufficient. The shoulder clod consists of multiple muscles and if it is not possible to declare that samples were consistently taken from the same location, the whole research is meaningless.

A1: We completely agree with your observation. A similar study was performed exclusively for texture from similar muscles (Ellies-Oury et al., 2019). In our study, the muscles from the forequarter (shoulder clod) were mostly M. triceps branchii, M. deltoideus and M. supraspinatus; and the meat pieces collected from the hindquarter (silverside) were a mixture of M. gluteobiceps, M. tensor fasciae latae and M. lateral vastus. This information was added in the manuscript.

Q2: I believe that an analysis based on the calculation of correlation coefficients is not entirely appropriate. Firstly, the number of observations in some groups is insufficient, and secondly correlation does not imply causation. More appropriate statistical analysis options are available.

A2: Our statistical model is based on solid comparison of three different factors on meat quality. The results obtained from the MANOVA clearly show that some factors, or their interactions play an important role in meat quality. The statistical MANOVA test can be used to examine significant differences between groups in a combination of dependent variables. By looking at several dependent variables at the same time, the MANOVA method enables a more comprehensive understanding of group differences and patterns. However, we know that there are different perspectives on how to analyze the data, but in our point of view, this is the most suitable one.

Values from Pearson and Spearman correlations were very similar. However, just to be in the safe side, we also consider that when calculating the Spearman nonparametric coefficients, this test is independent from normality infractions that may have occurred in the model.

Q3: 2.4 Sensory analysis: The method of storing meat for sensory analysis is quite strange and quite easily allows inconsistent treatment of all samples equally.

A3: Thank you for your comment. It is clear that freezing has an influence on the samples compared to fresh samples. For this reason, all samples were frozen until analysis. In fact, the treatment of all steaks was driven in such a way that minimal differences existed between working weeks. The aim of freezing the samples was precisely to standardize all the timings for sensory evaluation and WBSF determination.

Q4: It is necessary to indicate how the descriptors were assessed. It is necessary to indicate the scale; it is not obvious at first sight whether a higher value is more favourable or vice versa.

A4: We agree on this observation. Taste, smell and hardness were measured using the punctuation from a 1 to 100 scale, being in all cases 1 for “not perceptible” and 100 for “intense”. The scale on how the sensory evaluation was conducted was added in the text.

Q5: Can you indicate how many replicate samples overall and for each treatment were evaluated?

A5: Here is the calculation:

In a working week, we used 10 judges to evaluate the sensorial characteristics of meat.

Every judge received 4 samples in a single sensory analysis, which means that 40 replicates were prepared for every treatment. In total, we evaluated the meat traits across 4 working weeks. This means that overall, we used 160 samples. These numbers were added in the text.  

Q6: 2.5 Shear force: This is a rather unusual combination in which the samples were evaluated. The WBSF is usually used to measure intact lean muscle mass. For minced meat, TPA is recommended. Explain why you have applied this procedure. It is then difficult to compare values with the cited literature, which takes a different approach. Describe better the dimensions of the cut samples.

A6: Dear reviewer, we agree with this affirmation. Some authors mention that TPA is a better approach to measure the texture characteristics of meat. However, some authors also mention that the WBSF method is better when analyzing cooked meat samples (de Huidobro et al., 2005; Novaković & Tomašević, 2017), which was our case. The dimensions of the samples were the same as descripted in the sensory analysis. A more detailed explanation was added in the paper.

Q7: 2.6 Statistical analysis: The statistical model for sensory analysis should account for the judge effect.

A7: In our experiment, we had a panellist replication, this is when panellist evaluate the same product in different days, and the main objective of the trained panel was to obtain information from different treatments of beef. That is why the judge effect was rather treated as a fixed term than a random one (Lundahl & McDaniel, 1988).

It was also stated that judges might lose their ability to discriminate samples when 6 or more samples are evaluated per session. In our study, “the judge effect” did not count, since the sample size analysed per judge in every single sensorial session was never higher than four (Avery & Masters, 1999).

Q8: There is no mention of how the correlations are calculated.

A8: Correlations were found using a MANOVA model, which was explained in the manuscript in section 3.1.  But in general, correlations between factors were obtained using a Pillay’s Trace test. This information was already explained in point 3.9, but it will be added to the statistical model description as suggested by the reviewer.

Q9: Specific comments L47: hardness !

A9: Ups! It looks like we missed that one. Thank you very much for letting us know.

Q10: L54: A significantly longer time interval than 7 to 21 days is recommended to achieve the favourable organoleptic characteristics of dry-aged meat. This is also stated in the literature cited.

A10: Thank you for your comment. In our case, the meat pieces were not dry-aged, but wet-aged. That is why we took in consideration a maturing time of two weeks, which is into the span of 7 to 21d.

Q11: L141: Justify why blooming time 0 was applied? The is not the usual way of measuring the colour of beef.

A11: We appreciate your comment and have added more details. We totally agree on this point with you. In our case, the blooming time was at least 1h at 2°C before color measuring for all meat pieces as described in line 135.

Q12: L170: change WSBF to WBSF (to keep it consistent throughout the text)

A12: We are sorry about this typing mistake. We changed that immediately.

Q13: L424: I don't think it is appropriate to refer to silverside as low value cuts. It may not be suitable for steak preparation, but the main muscle (BF) is often described as economically important or valuable. Please rephrase that sentence.

A13: Dear reviewer. Thank you for this input. This is not what we meant to say. Of course, silverside and shoulder clod cuts can be used for different applications in HoReCa, fast food and they are very appreciated in the meat industry as raw material for Goulash and Rouladen (at least here in Germany). We referred to low value cuts, because some experts in the field have made this differentiation between muscles used for locomotion, which may have higher problems with color during wet-aging, due to higher myoglobin and mitochondrial content (Bekhit et al., 2019; Ramanathan et al., 2020). We appreciate your comment, and we changed that in the text.  

 

• PD: In line 163, there was a small mistake. The samples were cooked until a core temperature of 72°C and not 75° C. We apologize for this error.

 

References:

Avery, P. J., & Masters, G. A. (1999). Advice on the Design and Analysis of Sensory Evaluation Experiments Using a Case-study with Cooked Pork. Journal of the Royal Statistical Society: Series D (The Statistician), 48(3), 349-359. doi:https://doi.org/10.1111/1467-9884.00193

Bekhit, A., Morton, J., Bhat, Z., & Kong, L. M. (2019). Meat colour: factors affecting colour stability. In Encyclopedia of Food Chemistry (Vol. 2, pp. 202-210): Academic Press, Elsevier Publications

de Huidobro, F. R., Miguel, E., Blázquez, B., & Onega, E. (2005). A comparison between two methods (Warner–Bratzler and texture profile analysis) for testing either raw meat or cooked meat. Meat Science, 69(3), 527-536. doi:https://doi.org/10.1016/j.meatsci.2004.09.008

Ellies-Oury, M.-P., Lorenzo, H., Denoyelle, C., Saracco, J., & Picard, B. (2019). An Original Methodology for the Selection of Biomarkers of Tenderness in Five Different Muscles. Foods, 8(6), 206. Retrieved from https://www.mdpi.com/2304-8158/8/6/206

Lundahl, D. S., & McDaniel, M. R. (1988). The panelsit effect- fixed or random? . Journal of Sensory Studies, 3(2), 113-121. doi:https://doi.org/10.1111/j.1745-459X.1988.tb00434.x

Novaković, S., & Tomašević, I. (2017). A comparison between Warner-Bratzler shear force measurement and texture profile analysis of meat and meat products: a review. IOP Conference Series: Earth and Environmental Science, 85(1), 012063. doi:10.1088/1755-1315/85/1/012063

Ramanathan, R., Kiyimba, F., Gonzalez, J., Mafi, G., & DeSilva, U. (2020). Impact of up- and downregulation of metabolites and mitochondrial content on pH and color of the longissimus muscle from normal-pH and dark-cutting beef. J. Agric. Food Chem., 68(27), 7194-7203. doi:10.1021/acs.jafc.0c01884

Author Response File: Author Response.docx

Reviewer 2 Report

Comments and Suggestions for Authors

It is an interesting short paper, not so much for the novelty of the subject but because of its potential contribution to better evaluation and understanding of the factors affecting meat quality. However it needs major changes before being ready for publication. 

In terms of structure, as it is, it seems that lairage, mixing of unfamiliar animals and slaughtering temperature were treatments studied by the authors, as are transport distance (TD), animal weight (AW), and muscle position (MP). However, if that was the case, no data are presented to support that idea. In fact, about the former factors, the authors simply indicate the conditions verified during the study, without showing any proper analysis. The authors, themselves start the abstract saying “This study investigated the potential effects of transport distance, animal weight and muscle position on meat quality, under commercial conditions across four slaughtering weeks during the summer months (May to September). Data on transport, lairage and slaughtering were collected” So, data on transport, lairage and slaughtering should be presented in “Materials and Methods” and not in “Results and Discussion”. Also, it is strange to call Table 6, before calling (and presenting) Tables 3, 4 and 5. It is questionable, but if the authors want to start the “Results and Discussion” with an overall analysis (in line with the way they start the “Conclusions”), maybe it would be better to eliminate section “3.9. Multivariate test analysis of factor combination on meat quality traits” and include its contents in section 3.1, renumbering Table 6 to Table 3 (and correcting the number of Tables 3, 4 and 5). Also, the title of section 3.1 could be changed to something like “3.1. Combined effects of transport distance (TD), animal weight (AW), and muscle position (MP) on meat quality traits" since sections 3.3, 3.4 and 3.6 deal with the effect of each factor separately...

Besides these general remarks:

LINE 18 – Is it really necessary to put ΔE) here?

 

LINES 78-79 – The authors mention “For this reason, the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed.”. Throughout the paper there are several references to temperature, and temperature interval, but how was the temperature effect analysed?

LINES 79-82 – The authors mention “To find possible interactions between instrumental and sensory texture analysis and meat quality, Warner-Bratzler shear force (WBSF) and sensory analysis from cooked meat balls of the same meat cuts were additionally evaluated.”. Is “interactions” what the authors really want to say? If so, please explain.

LINES 207-209 – In Table 3, although it is a common notation, the meaning of different (or not) superscripts should be explained...

 

LINES 286-287 – The authors say “The meat from animals of short TD also exhibited a softer meat texture than the other groups, as determined through instrumental measurements and sensory evaluation.” What data support this? were the differences between TD treatments significant?

 

LINES 303-305 – In Table 5, What do "Smell %", "Taste %" and "Texture %" mean? Since the authors make a parallel between WBSF values and sensory tenderness values (e.g., lines 355-366), how do they compare WBSF (%) with Texture (%)?

 

LINES 321-323 – The authors say “Short processing times were reflected in superior meat quality, due to pH values lower than 5.8 in all treatment combinations.“ . What data led the authors to conclude that the lower than 5.8 pH values observed were due to short processing?

 

LINES 333-334 – The authors say “It was evidenced from our studies that high AW treatments resulted in lower pH values“ . What data support this "evidence"?

 

LINES 339-340 – The authors say “Indeed, slightly increased L* values were found in the meat from high AW treatments.”. Was there a significant difference between AW treatments or is just a trend? The authors should make this clear, since from the data shown no clear conclusion seems to be possible.

 

LINES 341-343 – The authors say “Increased tenderness could also be found from high AW combinations from both WBSF measurements and from the consumer’s point of view”. What data support this? Were the differences between TD treatments significant?

 

LINES 363-364 – The authors say “The SC muscles showed a higher ultimate pH than the SS, which subsequently resulted in improved color readings (L*, a* and b*) than SS.”. The authors had already "touched" this somehow (lines 227-230 and 233-235) when mentioning the correlations. Anyway, put this way, it seems that MP had a significant effect on all these parameters...

 

LINES 364-366 – The authors say “Interestingly, higher pH values in the SC were not correlated neither with WBSF values or with sensory tenderness values and showed that SC were rated as more tender than SS.”. Again - were all these differences significant? What data support such conclusion? Another question - the samples tested were from fried meatballs made with minced meat obtained from SC and SS and not actual samples of meat slices from SC and SS. Right? While that is mentioned in "Introduction" and "Materials and Methods" the way it is presented here may induce the reader in error...  finally - along the paper the authors mix "texture" and "tenderness", and make a parallel between "WBSF values" and "sensory tenderness values"...  While  many papers don't distinguish between "texture" and "tenderness" the fact is that texture of meat refers to firmness (toughness or degree of tenderness), cohesiveness and juiciness. I would sugest the authhors clearly state what they analysed (texture or tenderness), avoiding to mention sometimes "tenderness" and other times "texture", throughout the paper.

 

LINES 368-370 – The authors say “Taste and smell values were also higher for SC than SS and reaffirmed that in this case, these sensory traits were not related to pH development in meat.”. Again - were these differences significant? What data support such conclusion? What is the scale of these "taste and smaell values"

 

LINES 381-383 – The authors say “In the present study, cattle were carefully transported to the abattoir in small lots of familiar animals that indicated lower pre-mortem stress and higher meat quality attributes.” What data "indicated" lower pre-mortem stress and higher meat quality attributes?

 

LINES 384-385 – The authors say “stress-related problems did not seriously affect the physiological status of the young bulls.”. What data allow the authors conclude this?

 

LINES 399-401 – The authors say “This resulted in neglectable animal stress caused by weather fluctuations, due to rather mild temperatures. Animals were not exposed to any important temperature-related stress factors that could significantly affect the meat quality (Table 1).” As the authors put this, it seems they studied the effect of temperature,.. Was that the case? Maybe this coulld be simply changed to "this means that the animals were not exposed to any important temperature-related stress factors that could significantly affect the meat quality (Table 1)." 

 

LINES 419-422 – The authors say “Improved meat quality traits could be obtained from animals that had the shortest transport journeys (TD<34.5km) and the highest animal weight (AW>445). It seemed that the combination of these factors could have been beneficial in developing acceptable color and texture characteristics from both shoulder clod and silverside.”. However, considering just TD and AW, only pH48, L* and b* showed sugnificant correlations, and just with TD. Can you take such a clear conclusion? While, overall this is true, it seems an excessively generic conclusion...

 

LINES 424-427 – The authors say “Mild temperatures during the slaugh-424 tering days between May and September and short lairage times (<2 hours) did not im-425 portantly affect the physiological status of the young bulls and might have also contrib-426 uted to improve the meat quality parameters.”. As mentioned before - where is the data supporting this?

 

LINES 430-431 – The authors say “higher pH48 values were obtained from the shoulder clod, which resulted in lower L*, a* and b* readings.”. Why do the authors conclude that lower L*, a* and b* readings were consequence of higher pH48 values?

 

LINES 434-437 – The authors say “Future investigations should consider other year seasons and more extreme temperatures to find if variable weather conditions exert a more drastic influence on the meat quality of specific cuts such 436 as shoulder-clod and silverside.”. Please see above note about the temperature effect.

 

LINE 568 – Instead of “Bekhit, A.; Morton, J.; Bhat, Z.; Kong, L.M. Meat colour: factors affecting colour stability. 2019; pp. 202-210.“ should be “Bekhit, A.; Morton, J.; Bhat, Z.; Kong, L.M. Meat colour: factors affecting colour stability. In Encyclopedia of Food Chemistry, Volume 2, Academic Press, Elsevier Publications, 2019 pp.202-210, doi:10.1016/B978-0-12-814026-0.21665-X”.

Comments on the Quality of English Language

LINE 34 – Please change “for an specific” to “for a specific”

LINE 40 – Please change “(redness), and” to “(redness) and”

 

LINE 45 – Please change “role in opinion of consumers” to “role on the opinion of consumers”.

LINE 49 – Please change “meat could be” to “meat can be”.

LINE 51 – Please change “tenderness could be” to tenderness can be”.

LINES 53-54 – Please change “between -1 and 2°C, for around 7 and 21 days [18,19].” to “between -1 and 2°C [18], for around 7 and 21 days [19].”.

LINES 54-55 – Please change “more efficient, and safer” to “more efficient and safer”.

LINE 65 – Please change “might” to “may”.

LINE 69 – Please change “might” to “may”.

LINES 91-92 – Please change “600 until 1800 h” to “6:00 until 18:00 h”.

LINE 135 – Please change “Then instrumental” to “Then, instrumental”.

 

LINES 135 AND 163 – Please change “Then” to “Then,”.

 

LINE 167 – You should't begin a sentence with a numeral. Please change “10 technical replicates” to “Ten technical replicates”.

 

LINE 183 – Please change “(MP) and” to “(MP), and”.

 

LINE 184 – Please change “divided in” to “divided into”.

 

LINES 186 AND 187 – In “heavy animals Leip(AW>445 kg)” what is “Leip”?

 

LINE 195 – Please change “weight (AW), and muscle” to “weight (AW) and muscle”.

 

LINE 203 – Please change “a significant effects” to “significant effects” or to “a significant effect”

 

LINE 225 – Please change “correlations between TD, AW and MP on meat quality” to something like “correlations of TD, AW and MP with meat quality traits”.

 

LINES 228-230 – Please change “This suggested that with increasing TD, it will cause pH48 and L* values to increase and b* values to decrease.” to something like “This suggests that increasing TD will cause pH48 and L* values to increase, and b* values to decrease.".

 

LINES 233-235 – Please change “Since the numerical interpretation for MP in the MANOVA model was SC=1 and SS=2, a significant negative relationship was found between MP and pH48, which showed that decreasing pH48 could be significantly found from silverside steaks.” to something like “Since the numerical notation for MP in the MANOVA model was SC=1 and SS=2, the significant negative relationship found between MP and pH48 means that a significant lower pH48 could be found for silverside steaks.".

 

LINES 270-273 – The sentence “For instance, cattle transported in extreme long journeys (up to 1800 km) supposed a high strain in the animals and the probability to find DFD-related problems greatly increased, especially when lairage time was insufficient for the animals to recover after long journeys [34,35].” needs rephrasing. I assume "supposed" is not the verb you want to use...

 

LINES 273-274 – The sentence “Other study confirmed that bulls transported long distances (300 km) 273 resulted in significantly higher meat pH than bulls transported for only 125 km [36].” needs rephrasing. Maybe to something like "Other study confirmed that bulls transported long distances (300 km resulted in significantly higher meat pH than in bulls transported for only 125 km [36]."

 

LINE 277 – Please change “transported for medium distances” to “transported over medium distances”.

 

LINE 283 – Please change “previous opinion that referred” to “previous opinion”.

 

LINE 365 – Please change “neither with WBSF values or” to “neither with WBSF values nor”.

 

LINE 378 – Please change “output, and” to “output and”.

 

LINE 380 – Please change “transporting, and” to “transporting and”.

 

LINE 387 – Please change “precipitations, and” to “precipitations and”.

Author Response

Response to reviewer 2

 

It is an interesting short paper, not so much for the novelty of the subject but because of its potential contribution to better evaluation and understanding of the factors affecting meat quality. However, it needs major changes before being ready for publication. 

Thank you very much for the constructive comments about our research. We were very pleased to have you as our reviewer and to discuss about your point of view about our work. We tried to follow your suggestions to enhance the quality of our paper.

 

Q1: In terms of structure, as it is, it seems that lairage, mixing of unfamiliar animals and slaughtering temperature were treatments studied by the authors, as are transport distance (TD), animal weight (AW), and muscle position (MP). However, if that was the case, no data are presented to support that idea. In fact, about the former factors, the authors simply indicate the conditions verified during the study, without showing any proper analysis. The authors, themselves start the abstract saying “This study investigated the potential effects of transport distance, animal weight and muscle position on meat quality, under commercial conditions across four slaughtering weeks during the summer months (May to September). Data on transport, lairage and slaughtering were collected” So, data on transport, lairage and slaughtering should be presented in “Materials and Methods” and not in “Results and Discussion”. Also, it is strange to call Table 6, before calling (and presenting) Tables 3, 4 and 5. It is questionable, but if the authors want to start the “Results and Discussion” with an overall analysis (in line with the way they start the “Conclusions”), maybe it would be better to eliminate section “3.9. Multivariate test analysis of factor combination on meat quality traits” and include its contents in section 3.1, renumbering Table 6 to Table 3 (and correcting the number of Tables 3, 4 and 5). Also, the title of section 3.1 could be changed to something like “3.1. Combined effects of transport distance (TD), animal weight (AW), and muscle position (MP) on meat quality traits" since sections 3.3, 3.4 and 3.6 deal with the effect of each factor separately...

A1: Dear reviewer, thank you very much for your suggestions. We moved section 3.9 to 3.1 and numerated all tables as you recommended.

The data of transport, slaughtering and lairage were indeed presented in Tables 1 and 2 in the materials and methods section. The discussion of these two factors in the results and discussion section was interpreted according to the information obtained from the statistical model and references were cited to provide a background for this purpose.

 

Besides these general remarks:

 

 

Q2: LINE 18 – Is it really necessary to put ΔE here?

A2: It has been stated that color is one of the most important parameters in meat quality, from a consumer’s perspective (Tomasevic et al., 2021). The importance of ∆E in the present study is crucial, since the human eye can process color as a tridimensional stimuli (Hernández et al., 2019). Therefore, these values are indeed very important to compare the total color difference between shoulder clod and silverside and to determine if consumers can visually predict which muscles belong to each muscle position. However, we agree with you that this content cut the flow of the ideas in the abstract. Therefore, we changed it at the end of the section to increase the clarity of the ideas. We also added a couple of sentences to enhance the context the text.

Q3: LINES 78-79 – The authors mention “For this reason, the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed.”. Throughout the paper there are several references to temperature, and temperature interval, but how was the temperature effect analysed?

A3: Thank you for pointing out this issue. In this sentence in specific we refer to meat temperature because pH post-mortem decline is closely related to this parameter. In some other parts of the text, the word “temperature” is referred to air temperature during slaughtering, which can also have a deep effect on meat quality (Poveda-Arteaga et al., 2023).  

Q4: LINES 79-82 – The authors mention “To find possible interactions between instrumental and sensory texture analysis and meat quality, Warner-Bratzler shear force (WBSF) and sensory analysis from cooked meat balls of the same meat cuts were additionally evaluated.”. Is “interactions” what the authors really want to say? If so, please explain

A4: Thanks for clearing this point out. You are right, maybe “interaction” is not precisely the term we wanted to use. We wanted to find comparisons between these two measurements and to see which one may interpret better the results of our study. This word was changed in the text for the word “comparison”.

Q5: LINES 207-209 – In Table 3, although it is a common notation, the meaning of different (or not) superscripts should be explained...

A5: We agree on that. We changed this as indicated.

 

Q6: LINES 286-287 – The authors say “The meat from animals of short TD also exhibited a softer meat texture than the other groups, as determined through instrumental measurements and sensory evaluation.” What data support this? were the differences between TD treatments significant?

A6: This is correct. This information was taken from table 5. We also incorporated the superscripts for TD to differentiate between significant and non-significant values.

 

Q7: LINES 303-305 – In Table 5, What do "Smell %", "Taste %" and "Texture %" mean? Since the authors make a parallel between WBSF values and sensory tenderness values (e.g., lines 355-366), how do they compare WBSF (%) with Texture (%)?

A7: Meat taste, smell and hardness were measured using the punctuation of a 1 to 100 scale, being in all cases 1 for “not perceptible” and 100 for “intense”. This description was added in the text and in the table explanation. Also, the percentage symbol was erased, since this measurements considered a scale and not a percentage per se.

 

From our point of view, the sensory evaluation of meat texture is proportionally related with the instrumental measurement of shear force. That is why we used this correlation to describe meat quality. 

 

Q8: LINES 321-323 – The authors say “Short processing times were reflected in superior meat quality, due to pH values lower than 5.8 in all treatment combinations.“ . What data led the authors to conclude that the lower than 5.8 pH values observed were due to short processing?

A8: It has been mentioned in the literature that when beef attains pH values lower than around 5.8, then it is considered to have improved texture and color than meat from higher pH values, which falls into the DFD status (Lomiwes et al., 2013; McKeith et al., 2016; Page et al., 2001). We concluded that these relatively normal pH₄₈ values from all treatments are linked to fast processing times that we saw in our experiments (Table 2), also because we did not find any important color defects in any of our samples.

 

Q9: LINES 333-334 – The authors say “It was evidenced from our studies that high AW treatments resulted in lower pH values“ . What data support this "evidence"?

A9: The information presented in Table 5 supports that heavier animals exhibit a trend for lower meat pH values. We have incorporated this reference into the text. 

 

Q 10: LINES 339-340 – The authors say “Indeed, slightly increased L* values were found in the meat from high AW treatments.”. Was there a significant difference between AW treatments or is just a trend? The authors should make this clear, since from the data shown no clear conclusion seems to be possible.

A10: This is just a trend. We could not find any significant differences for AW treatments by using a post-hoc test, since this factor was only divided into light (AW<445kg) and heavy (AW>445kg) animals. Also, according to the Spearman correlations calculated in Table 4, there was not any significant correlations between AW and any of the variables.

 

Q11: LINES 341-343 – The authors say “Increased tenderness could also be found from high AW combinations from both WBSF measurements and from the consumer’s point of view”. What data support this? Were the differences between TD treatments significant?

A11: This conclusion is supported by table 5. As in the previous question, we can only imply these results as a trend and significance does not apply in this example, because this factor was only divided into two groups.

 

Q12: LINES 363-364 – The authors say “The SC muscles showed a higher ultimate pH than the SS, which subsequently resulted in improved color readings (L*, a* and b*) than SS.”. The authors had already "touched" this somehow (lines 227-230 and 233-235) when mentioning the correlations. Anyway, put this way, it seems that MP had a significant effect on all these parameters...

A 12: This information is correct. This comparison was made when writing about the Pearson correlations between main factors and variables and indicated that there was a negative significant correlation between muscle position and pH₄₈. However, we cannot say that this was a significant correlation, also because the factor MP was designed to have only in two levels.

Values from Pearson and Spearman correlations were very similar. However, just to be in the safe side, we also consider that when calculating the Spearman nonparametric coefficients, this test is independent from normality infractions that may have occurred in the model.

 

Q 13: LINES 364-366 – The authors say “Interestingly, higher pH values in the SC were not correlated neither with WBSF values or with sensory tenderness values and showed that SC were rated as more tender than SS.”. Again - were all these differences significant? What data support such conclusion? Another question - the samples tested were from fried meatballs made with minced meat obtained from SC and SS and not actual samples of meat slices from SC and SS. Right? While that is mentioned in "Introduction" and "Materials and Methods" the way it is presented here may induce the reader in error...  finally - along the paper the authors mix "texture" and "tenderness", and make a parallel between "WBSF values" and "sensory tenderness values"...  While  many papers don't distinguish between "texture" and "tenderness" the fact is that texture of meat refers to firmness (toughness or degree of tenderness), cohesiveness and juiciness. I would suggest the authors clearly state what they analysed (texture or tenderness), avoiding to mention sometimes "tenderness" and other times "texture", throughout the paper.

A 13 :Thank you bringing this matter into our attention. It has been previously published that problems associated with high pH meat can also lead to harder texture (Ponnampalam et al., 2017). Therefore, we expected to obtain tougher meat from SC, because it had higher pH values than SS as shown in Table 5. However, our data suggested that WBSF values were higher in SS and not in SC. From a sensory perspective, same correlations were obtained and showed that SS was tougher than SC, despite of having lower pH. No post-hoc test is available, since there are only two means, and the information can only be compared, but no significance level can be calculated.    

 

For your second question, the answer is yes, this is right. Texture analysis was in both cases made from minced meat and not from the actual steaks. The reason why we designed the experiment in this way is because we were aiming to have more uniform samples. If single steaks would have been taken for every measurement, this would have taken much more time and probably less reliable results could have been obtained. Also, different steaks had also different fat contents, which would have affected the clarity of the results. Therefore, steaks were trimmed from the fat before mincing the meat.   

 

And for the third point, we also agree with your opinion. Texture is a wider quality attribute in meat. However, it is difficult to just attach to only one of the two terminologies since the development of the ideas sometimes need one or the other. We tried to keep it as straight forward as possible to prevent the indiscriminate use of those two words.

 

Q 14: LINES 368-370 – The authors say “Taste and smell values were also higher for SC than SS and reaffirmed that in this case, these sensory traits were not related to pH development in meat.”. Again - were these differences significant? What data support such conclusion? What is the scale of these "taste and smaell values"

A 14: Taste, smell and hardness were measured using the punctuation from a 1 to 100 scale, being in all cases 1 for “not perceptible” and 100 for “intense”. The idea of this sentence was to emphasize that even if higher values for smell and taste were expected from SS, which is an indicator for superior meat quality, exactly the contrary occurred. 

 

Q 15: LINES 381-383 – The authors say “In the present study, cattle were carefully transported to the abattoir in small lots of familiar animals that indicated lower pre-mortem stress and higher meat quality attributes.” What data "indicated" lower pre-mortem stress and higher meat quality attributes?

A 15: From our point of view, lower pre-mortem stress was reflected in overall low pH values of all meat pieces studied. The data presented in Table 5 indicated that any of the treatments had pH values higher than 5.8, which was determinant in enhancing meat quality (Poveda-Arteaga et al., 2023; Węglarz, 2010). Other studies indicated that higher pH values can be obtained from cattle that had longer transportation times before arriving to the abattoir (Romero et al., 2013). Also, the fact that ante-mortem stress increases when cattle are mixed with unfamiliar animals on their way to the slaughterhouse (Mounier et al., 2006).

In our case, animal stress was reduced by transporting only familiar animals, which were separated in the pins before slaughtering.

 

Q 16: LINES 384-385 – The authors say “stress-related problems did not seriously affect the physiological status of the young bulls.”. What data allow the authors conclude this?

A 16: As previously described, this information was concluded based on observed pH values of all treatments collected in this experiment ranging between 5.47 to 5.78.

 

Q 17: LINES 399-401 – The authors say “This resulted in neglectable animal stress caused by weather fluctuations, due to rather mild temperatures. Animals were not exposed to any important temperature-related stress factors that could significantly affect the meat quality (Table 1).” As the authors put this, it seems they studied the effect of temperature,.. Was that the case? Maybe this coulld be simply changed to "this means that the animals were not exposed to any important temperature-related stress factors that could significantly affect the meat quality (Table 1)." 

A 18: Thank you for this important input. This is exactly what we wanted to express. This information was changed in the paper as you suggested. In this case, the environmental temperature was one of the factors that contributed to higher meat quality, but it was not taken in consideration in the statistical model.

 

Q 19: LINES 419-422 – The authors say “Improved meat quality traits could be obtained from animals that had the shortest transport journeys (TD<34.5km) and the highest animal weight (AW>445). It seemed that the combination of these factors could have been beneficial in developing acceptable color and texture characteristics from both shoulder clod and silverside.”. However, considering just TD and AW, only pH48, L* and b* showed significant correlations, and just with TD. Can you take such a clear conclusion? While, overall this is true, it seems an excessively generic conclusion...

A 19: We appreciate your comment, however, we do not consider that this is quite a general conclusion, but we consider that this affirmation is true for our study. We tried to paraphrase the sentence, in a way that it sounds more related to this study only.

 

Q 20: LINES 424-427 – The authors say “Mild temperatures during the slaughtering days between May and September and short lairage times (<2 hours) did not importantly affect the physiological status of the young bulls and might have also contributed to improve the meat quality parameters.”. As mentioned before - where is the data supporting this?

A 20: This data can be obtained from Table 2, which has data concerning air temperature and lairage times for all animals from this study. We added this reference in the text as suggested.

 

Q 21: LINES 430-431 – The authors say “higher pH48 values were obtained from the shoulder clod, which resulted in lower L*, a* and b* readings.”. Why do the authors conclude that lower L*, a* and b* readings were consequence of higher pH48 values?

A 21: Several reviews in the field suggested that high pH values are directly related to dark meat colors, a defect which is better known as DFD meat (Neethling et al., 2017; Poveda-Arteaga et al., 2023; Ruedt et al., 2023; Tomasevic et al., 2021). A combination of factors may increase the probability to find these problems in beef and animal stress is recognized as one of the most important reasons why this issue may occur. Therefore, we concluded that as pH values get closer to 5.6, then the color coordinates of the beef pieces in our experiment will also increase as shown in Table 5.  

 

Q22: LINES 434-437 – The authors say “Future investigations should consider other year seasons and more extreme temperatures to find if variable weather conditions exert a more drastic influence on the meat quality of specific cuts such as shoulder-clod and silverside.”. Please see above note about the temperature effect.

A 22: We speculate that this may be an interesting topic for future investigations. Because animals were not exposed to extreme low or high temperatures in our study, we assume that this should be investigated over a longer time span.

 

Q23: LINE 568 – Instead of “Bekhit, A.; Morton, J.; Bhat, Z.; Kong, L.M. Meat colour: factors affecting colour stability. 2019; pp. 202-210.“ should be “Bekhit, A.; Morton, J.; Bhat, Z.; Kong, L.M. Meat colour: factors affecting colour stability. In Encyclopedia of Food Chemistry, Volume 2, Academic Press, Elsevier Publications, 2019 pp.202-210, doi:10.1016/B978-0-12-814026-0.21665-X”.

A 24: Thank you for your suggestion. The bibliography was automatically created by EndNote, but we solved this editing problem.

 

Comments on the Quality of English Language

LINE 34 – Please change “for an specific” to “for a specific”

It was changed as suggested.

LINE 40 – Please change “(redness), and” to “(redness) and”

It was changed as suggested.

LINE 45 – Please change “role in opinion of consumers” to “role on the opinion of consumers”.

It was changed as suggested.

LINE 49 – Please change “meat could be” to “meat can be”.

It was changed as suggested.

LINE 51 – Please change “tenderness could be” to tenderness can be”.

It was changed as suggested.

LINES 53-54 – Please change “between -1 and 2°C, for around 7 and 21 days [18,19].” to “between -1 and 2°C [18], for around 7 and 21 days [19].”

It was changed as suggested.

LINES 54-55 – Please change “more efficient, and safer” to “more efficient and safer”.

It was changed as suggested.

LINE 65 – Please change “might” to “may”.

It was changed as suggested.

LINE 69 – Please change “might” to “may”.

It was changed as suggested.

LINES 91-92 – Please change “600 until 1800 h” to “6:00 until 18:00 h”.

It was changed as suggested.

LINE 135 – Please change “Then instrumental” to “Then, instrumental”.

It was changed as suggested. 

LINES 135 AND 163 – Please change “Then” to “Then,”.

It was changed as suggested. 

LINE 167 – You should't begin a sentence with a numeral. Please change “10 technical replicates” to “Ten technical replicates”.

It was changed as suggested. 

LINE 183 – Please change “(MP) and” to “(MP), and”.

It was changed as suggested.

LINE 184 – Please change “divided in” to “divided into”.

It was changed as suggested.

LINES 186 AND 187 – In “heavy animals Leip(AW>445 kg)” what is “Leip”?

We are very sorry to find this error in the text. It has no meaning. We deleted it.

 

LINE 195 – Please change “weight (AW), and muscle” to “weight (AW) and muscle”.

It was changed as suggested.

LINE 203 – Please change “a significant effects” to “significant effects” or to “a significant effect”

It was changed as suggested.

 

LINE 225 – Please change “correlations between TD, AW and MP on meat quality” to something like “correlations of TD, AW and MP with meat quality traits”.

 It was changed as suggested.

LINES 228-230 – Please change “This suggested that with increasing TD, it will cause pH48 and L* values to increase and b* values to decrease.” to something like “This suggests that increasing TD will cause pH48 and L* values to increase, and b* values to decrease.".

 It was changed as suggested.

LINES 233-235 – Please change “Since the numerical interpretation for MP in the MANOVA model was SC=1 and SS=2, a significant negative relationship was found between MP and pH48, which showed that decreasing pH48 could be significantly found from silverside steaks.” to something like “Since the numerical notation for MP in the MANOVA model was SC=1 and SS=2, the significant negative relationship found between MP and pH48 means that a significant lower pH48 could be found for silverside steaks.".

It was changed as suggested.

LINES 270-273 – The sentence “For instance, cattle transported in extreme long journeys (up to 1800 km) supposed a high strain in the animals and the probability to find DFD-related problems greatly increased, especially when lairage time was insufficient for the animals to recover after long journeys [34,35].” needs rephrasing. I assume "supposed" is not the verb you want to use...

It was changed as suggested. 

LINES 273-274 – The sentence “Other study confirmed that bulls transported long distances (300 km) 273 resulted in significantly higher meat pH than bulls transported for only 125 km [36].” needs rephrasing. Maybe to something like "Other study confirmed that bulls transported long distances (300 km resulted in significantly higher meat pH than in bulls transported for only 125 km [36]."

It was changed as suggested.

LINE 277 – Please change “transported for medium distances” to “transported over medium distances”.

It was changed as suggested.

LINE 283 – Please change “previous opinion that referred” to “previous opinion”.

It was changed as suggested. 

LINE 365 – Please change “neither with WBSF values or” to “neither with WBSF values nor”.

 It was changed as suggested.

LINE 378 – Please change “output, and” to “output and”.

It was changed as suggested.

LINE 380 – Please change “transporting, and” to “transporting and”.

It was changed as suggested.

LINE 387 – Please change “precipitations, and” to “precipitations and”.

It was changed as suggested.

 

• Thank you for taking the time to review the language mistakes. We really appreciate this from you!
• PD: In line 163, there was a small mistake. The samples were cooked until a core temperature of 72°C and not 75° C. We apologize for this error.

 

References:

Hernández, B., Sáenz Gamasa, C., Diñeiro Rubial, J. M., & Alberdi Odriozola, C. (2019). CIELAB color paths during meat shelf life. Meat Sci., 157, 107889. doi:https://doi.org/10.1016/j.meatsci.2019.107889

Lomiwes, D., Farouk, M. M., Frost, D. A., Dobbie, P. M., & Young, O. A. (2013). Small heat shock proteins and toughness in intermediate pHu beef. Meat Sci, 95(3), 472-479. doi:10.1016/j.meatsci.2013.05.022

McKeith, R. O., King, D. A., Grayson, A. L., Shackelford, S. D., Gehring, K. B., Savell, J. W., & Wheeler, T. L. (2016). Mitochondrial abundance and efficiency contribute to lean color of dark cutting beef. Meat Science, 116, 165-173.

Mounier, L., Dubroeucq, H., Andanson, S., & Veissier, I. (2006). Variations in meat pH of beef bulls in relation to conditions of transfer to slaughter and previous history of the animals. J. Anim. Sci., 84(6), 1567-1576. doi:10.2527/2006.8461567x

Neethling, N., Suman, S., Sigge, G., & Hunt, M. C. (2017). Exogenous and endogenous factors influencing color of fresh meat from ungulates. Meat Muscle Biol., 1, 253. doi:10.22175/mmb2017.06.0032

Page, J. K., Wulf, D. M., & Schwotzer, T. R. (2001). A survey of beef muscle color and pH. Journal of Animal Science, 79(3), 678-687. doi:10.2527/2001.793678x

Ponnampalam, E., Hopkins, D., Bruce, H., Baldi, G., & Bekhit, A. (2017). Causes and Contributing Factors to “Dark Cutting” Meat: Current Trends and Future Directions: A Review. Comprehensive Reviews in Food Science and Food Safety, 16. doi:10.1111/1541-4337.12258

Poveda-Arteaga, A., Krell, J., Gibis, M., Heinz, V., Terjung, N., & Tomasevic, I. (2023). Intrinsic and Extrinsic Factors Affecting the Color of Fresh Beef Meat&mdash;Comprehensive Review. Applied Sciences, 13(7), 4382. Retrieved from https://www.mdpi.com/2076-3417/13/7/4382

Romero, M., Uribe-Velásquez, L. F., Sanchez, J., & Miranda-de la Lama, G. (2013). Risk factors influencing bruising and high muscle pH in Colombian cattle carcasses due to transport and pre-slaughter operations. Meat Sci., 95, 256-263. doi:10.1016/j.meatsci.2013.05.014

Ruedt, C., Gibis, M., & Weiss, J. (2023). Meat color and iridescence: Origin, analysis, and approaches to modulation. Compr. Rev. Food Sci. Food Saf., 22(4), 3366-3394. doi:https://doi.org/10.1111/1541-4337.13191

Tomasevic, I., Djekic, I., Font-i-Furnols, M., Terjung, N., & Lorenzo, J. M. (2021). Recent advances in meat color research. Current Opinion in Food Science, 41, 81-87. doi:https://doi.org/10.1016/j.cofs.2021.02.012

Węglarz, A. (2010). Meat quality defined based on pH and colour depending on cattle category and slaughter season. Czech J. Anim. Sci., 55, 548-556. doi:10.17221/2520-CJAS

 

 

Author Response File: Author Response.docx

Reviewer 3 Report

Comments and Suggestions for Authors

The study examined the potential effects of transport distance, animal weight, and muscle of young bulls on a number of meat quality attributes.  The study is fairly well designed and written, but some clarification and modifications of the methods and interpretation of the results are warranted. I’m curious why lairage time wasn’t investigated and discussed as thoroughly (at all) as so many other traits.  Please consider including in analysis.  Specific comments can be found below.

 

Ln14: specifically what data were collected? transport time and distance, lairage time, and what about slaughtering??

Ln22: patties not balls

Ln18, 42, and throughout: you’re not measuring a change, but simply a difference in color between two independent samples.  This is a valid way to report color values in this study. 

Ln47: hardness

Ln52: delete “so-called”

Ln81: patties not balls

Ln107: please clarify was each animal individually housed in a separate pen during lairage?

Ln109-113: what does EUROP mean?  What does carcass fat 1, 2, or 3 mean?  Please provide scales in text and footnotes for Table 1.  YOU also say that carcasses were classified by HCW in Table 1, but there are not weights in Table 1.

Ln115-116: specifically where and how were samples removed?  Was the 0.5 kg sample a single intact piece of muscle?  Was any attention paid to shape or dimension of the sample during removal? Please use scientific muscle names?  Were samples trimmed of external fat or connective tissue before packaging?

Ln119/133: see previous comment for new sample

Ln133-142: you do not mention anything about Delta E here.  I don’t see this as an appropriate way to report color in this study.  It was not a change in color.  You should report independent color values of the two muscles. 

Ln144: suggest using the word inserting rather than sticking

Ln150-165: I’m struggling to understand why samples were aged if you were just going to grind them up.  The shear force and sensory data would be so much more meaningful if you conducted the testing on whole muscle samples.  You essentially eliminated tenderness differences by grinding and introduced a pro-oxidative activity (grinding) to your samples. 

Ln163: please provide training protocols for panelists.  How many sessions were conducted?  How many samples per session?  Were samples only from a working week utilized since everything was kept frozen for 8 weeks?  How were samples portioned?  How were samples kept warm before serving?  How was order and presentation of samples balanced? How were samples coded to avoid bias?

Ln164: you must provide scales/anchors for sensory traits.

Ln165: change meat balls to meat patties

Ln167-172: please reference a shear force protocol for meat balls/patties.  Specifically how were samples portioned/cut prior to testing after cooking and how were samples oriented in the testing device?  Steaks would normally be cored parallel to fibers and sheared perpendicular to fibers.  However, there is no mention of sample orientation during this process. 

Ln173: I’m not seeing anything about Pearson correlation analysis, but I’m not convinced this is a useful statistical tool with meaningful results in this instance.

Ln203: effect not effects

Ln204: tables should be presented and discussed in order.  You have jumped from 3 to 6, but table 6 really appears much later on.

Table 3: Shear force and sensory were performed on cooked meat patties, yet pH and color testing was conducted on raw whole muscle samples (presumably).  This should be clearly delineated in the table description.   Why are sensory traits reported as percentages?  Percentages of what?  Scales used for sensory traits should be provided as footnotes to all tables. Define Delta E.  More importantly, you do not mention anything about Delta E in the methods.  I don’t see this as an appropriate way to report color in this study.  It was not a change in color.  You should report independent color values of the two muscles. 

Ln225-235: delete section and corresponding table.  There is no value.  Moreover, muscle position is categorical, so I have no idea how you managed to report something for muscle position in Table 4. 

Ln326-348: this entire section is based on a lot of assumptions.  Animals were not managed or fed the same or in a controlled environment.  Animal age could just as easily contribute to weight differences as differences in diet, which you have no idea what animals were fed.  At least you don’t share that information.  You didn’t quantify IMF so the discussion in this whole section is pretty far fetched for me.  I would delete or significantly revise based on what you actually know and what you actually tested and accounted for. 

Ln350: I’m struggling to see how you would see metmyoglobin accumulation in a wet aging environmental, especially if only for 2 weeks.  Wet aging is a no/low O2 environment and O2 is required for oxidation and MMb accumulation. 

Author Response

Response to reviewer 3

 

The study examined the potential effects of transport distance, animal weight, and muscle of young bulls on a number of meat quality attributes.  The study is fairly well designed and written, but some clarification and modifications of the methods and interpretation of the results are warranted.

Thank you very much for the constructive comments about our research. We were very pleased to have you as our reviewer and to discuss about your point of view about our work. We tried to follow your suggestions to enhance the quality of our paper.

 

Q1: I’m curious why lairage time wasn’t investigated and discussed as thoroughly (at all) as so many other traits.  Please consider including in analysis. Specific comments can be found below.

A 1: Thank you for this input. It would have been a good idea to include lairage time in our experimental design; however, we considered that the three factors studied in this manuscript (transport distance, animal weight and muscle position) had more relevant importance in beef quality.

 

Q2: Ln14: specifically what data were collected? transport time and distance, lairage time, and what about slaughtering??

A2: All the pre-mortem parameters that we could collect from the slaughterhouse about transport, lairage and slaughtering conditions of the animals were condensed and presented in tables 1 and 2. With slaughtering we mean the weather conditions on those days, which is an important parameter that can influence animal stress and meat quality.  

 

Q3: Ln22: patties not balls

A3: Thank you for this input. We changed it in the text as indicated.

 

Q4: Ln18, 42, and throughout: you’re not measuring a change, but simply a difference in color between two independent samples.  This is a valid way to report color values in this study. 

A4: Thank you for clearing this out. We totally agree with you, and we checked again the whole manuscript to make sure that this information was presented as you suggested.

 

Q5: Ln47: hardness

A5: Sorry about this typing mistake. We promptly corrected it.

 

Q6: Ln52: delete “so-called”

A6: Alright, we will proceed as suggested.

 

Q7: Ln81: patties not balls

A7: Corrected

 

Q8: Ln107: please clarify was each animal individually housed in a separate pen during lairage?

A8: They were not individually separated. They were only separated according to the truck they were coming from. This means in general terms, that animals coming from the same farm were also kept in the same group until slaughtering. 

 

Q9: Ln109-113: what does EUROP mean?  What does carcass fat 1, 2, or 3 mean?  Please provide scales in text and footnotes for Table 1.  YOU also say that carcasses were classified by HCW in Table 1, but there are not weights in Table 1.

A9: This is a very valid suggestion. Thank you for mentioning this point. We did exactly as suggested. You are right, HWC results were presented in Table 2 and not 1. We corrected this mistake too.

 

Q10: Ln115-116: specifically where and how were samples removed?  Was the 0.5 kg sample a single intact piece of muscle?  Was any attention paid to shape or dimension of the sample during removal? Please use scientific muscle names?  Were samples trimmed of external fat or connective tissue before packaging?

A10: The samples were obtained form the left side of the hanging carcasses once they entered the chilling rooms. The samples were in all cases obtained from a single piece of intact beef muscle, which was cut by slaughterhouse personnel. No especial attention was paid to the shape of the meat cut, but it was regularly a semi-circular one. The dimension of all samples was kept uniform, and it weighted around 0.5 kg. Samples were not trimmed immediately of neither of external fat nor connective tissue; however, they were trimmed of fatty tissue before mincing the meat for sensory analysis.  

 

A1: We completely agree with your observation. A similar study was performed exclusively for texture from similar muscles (Ellies-Oury et al., 2019). In our study, the muscles from the forequarter (shoulder clod) were mostly M. Triceps branchii, Deltoideus and Supraspinatus; and the meat pieces collected from the hindquarter (silverside) were a mixture of M. Gluteobiceps, M. Tensor fasciae latae and M. Lateral vastus. This information was added in the article.

 

Q11: Ln119/133: see previous comment for new sample

A11: Same standards were kept for obtaining the second sample for color measurement.

 

Q12: Ln133-142: you do not mention anything about Delta E here.  I don’t see this as an appropriate way to report color in this study.  It was not a change in color.  You should report independent color values of the two muscles. 

A12: It has been stated that color is one of the most important parameters in meat quality, from a consumer’s perspective (Tomasevic et al., 2021). The importance of ∆E in the present study is crucial since the human eye can process color as a tridimensional sensation (Hernández et al., 2019). Therefore, these values are indeed very important to compare the total color difference between shoulder clod and silverside and to determine if consumers can visually predict which muscles belong to each muscle position. However, individual color values of every treatment were also presented in a L*, a* and b* format as shown in Table 3.  

 

Q13: Ln144: suggest using the word inserting rather than sticking

A13: Thank you for your observation. It was changed as requested.

 

Q14: Ln150-165: I’m struggling to understand why samples were aged if you were just going to grind them up.  The shear force and sensory data would be so much more meaningful if you conducted the testing on whole muscle samples.  You essentially eliminated tenderness differences by grinding and introduced a pro-oxidative activity (grinding) to your samples. 

A14: Dear reviewer, the samples were aged for a two-week period, because we wanted to investigate if there were any color changes in the surface of the steaks during maturation.

 

Of course, grinding the muscles may increase the exposition of meat tissue to the action of oxidation groups. You are also right when you say that grinding the meat samples might decrease the tenderness differences between samples.

 

However, the idea of grinding the muscles before sensory analysis was to have a uniform sample that could be differentiated by the judges. It is also necessary to consider the excessive number of samples that would have been required to be processed and analyzed if samples of single muscles were presented to the judges. It was stated that judges might lose their ability to discriminate samples when 6 or more samples are evaluated per session. In our study, the sample size analysed per judge in every single sensorial session was never higher than four, which might highly increase the consistency of the results (Avery & Masters, 1999).

 

It was also a matter that if single muscles were used, probably we could not have enough material for sensory and shear force evaluations. Even if we tried to keep a uniform sample size and conformation, there were samples which were bigger or smaller than others, some other samples that had a thicker or finer fat layer and samples that had higher or lesser connective tissue. Thus, the objective of trimming the beef muscles before mincing the meat was also to have uniform samples for sensory and texture analysis.  

 

To prevent a high degree of oxidation in the meat samples, the mincing of the samples, forming of the meat patties and frying of the meat patties were strictly conducted at 4°C, and the sensory evaluation was performed on the same day.      

 

Q15: Ln163: please provide training protocols for panelists. How many sessions were conducted? 

A15: Thank you for these important comments. Before the initial sensory evaluation session, there was a brief introduction about the topic asked in the questionary and how meat traits should have been measured. The judges were selected from a group of workers at DIL that have previous experience in sensory analysis of meat samples. The questionary was self-explanatory. The sensory analysis was performed in four sessions in total, which is the number of working weeks that we got samples from the slaughterhouse.    

 

Q16: How many samples per session?  

A 16: Four samples were presented to the judges per session. This multiplied for the number of judges equals to 40 samples per session and 160 samples in total. 

 

Q17:Were samples only from a working week utilized since everything was kept frozen for 8 weeks?  

A 17:Yes, this is correct. Only samples from a single working week were presented in every sensory evaluation.

 

Q18: How were samples portioned?  How were samples kept warm before serving? How was order and presentation of samples balanced? How were samples coded to avoid bias?

A18: Samples were presented in paper cups covered with a lid, which preserved the heat and the smell from the fresh fried meat patties. Between cooking and presenting the samples to the judges there was only a 30-second period, and this is why there was no need to heat up the samples again after cooking. The samples were aleatory presented to the judges, with different identification numbers to reduce bias in our study as described in line 162.    

 

Q19: Ln164: you must provide scales/anchors for sensory traits.

A19: We agree with your opinion. Taste, smell and tenderness/hardness were measured using the punctuation of a 1 to 100 scale, being in all cases 1 for “not perceptible” and 100 for “intense”. This description was added in the text.

 

Q20: Ln165: change meat balls to meat patties

A20: We omitted this sentence to enhance clarity of the text, and we replace it in section 2.5. The term “meatballs” was changed for “meat patties” as suggested.

 

Q21: Ln167-172: please reference a shear force protocol for meat balls/patties.  Specifically how were samples portioned/cut prior to testing after cooking and how were samples oriented in the testing device?  Steaks would normally be cored parallel to fibers and sheared perpendicular to fibers.  However, there is no mention of sample orientation during this process. 

A21: We appreciate your suggestion, but we think that in this case the orientation of the fibers was irrelevant, since the shear force measurement was conducted on meat patties rather than directly in beef muscles. The reference for WBSF measurement was found and added to the text.

 

Q22: Ln173: I’m not seeing anything about Pearson correlation analysis, but I’m not convinced this is a useful statistical tool with meaningful results in this instance.

A22: We analyzed Pearson correlations in section 3.2. We believe that Pearson correlations explain important information about the factors studied in this manuscript. Values from Pearson and Spearman correlations were very similar. However, just to be in the safe side, we also consider that when calculating the Spearman nonparametric coefficients, this test is independent from normality infractions that may have occurred in the model. This was included in the final version of the paper.

 

Q23: Ln203: effect not effects

A23: Corrected

 

Q24:Ln204: tables should be presented and discussed in order.  You have jumped from 3

to 6, but table 6 really appears much later on.

A24: Thanks, you are right. We changed the order of the tables in the final version of the manuscript for better understanding of the ideas expressed in all sections.

 

Q25: Table 3: Shear force and sensory were performed on cooked meat patties, yet pH and color testing was conducted on raw whole muscle samples (presumably).  This should be clearly delineated in the table description.   Why are sensory traits reported as percentages?  Percentages of what?  Scales used for sensory traits should be provided as footnotes to all tables. Define Delta E.  More importantly, you do not mention anything about Delta E in the methods.  I don’t see this as an appropriate way to report color in this study.  It was not a change in color.  You should report independent color values of the two muscles. 

A25:Thank you for your suggestions. We agree with you, and we included the information that you proposed in the table description and deleted the percentage symbol from all the parameters from sensory analysis in all tables.

 

About the ∆E values, it has been published that it is a sensitive indicator to measure myoglobin oxidation changes in beef muscles (Hernández et al., 2019). However, in this paper the purpose of ∆E calculation was to differentiate the color of the two muscle positions and not color difference inside the same muscle. Therefore, we consider that the differences measured between SS and SC were valid, because they considered only individual L*, a* and b* coordinates from those independent muscles from table 3. The L*, a* and b* values from SS and SC are also presented in the same table.

 

Also, it is important to consider that ∆E calculation was used previously, for example, to compare color measurements between computer vision system (CVS) and a Minolta colorimeter (Tomasevic et al., 2019). In another manuscript, ∆E was also used to compare the total color difference between two muscles of several publications using the L*, a* and b* values from individual muscles (Poveda-Arteaga et al., 2023).

 

You are right when you say that the description of ∆E calculation was not added in the method section. We included this information in the manuscript, and we changed the order in which ∆E was presented in table 3 and we re-wrote it next to the color values.   

 

Q26: Ln225-235: delete section and corresponding table.  There is no value.  Moreover, muscle position is categorical, so I have no idea how you managed to report something for muscle position in Table 4.

A26: From all the parameters that we analyzed from the statistical model; muscle position was one of the most important. Even if muscle position is a categorical variable, when introduced in SPSS it can be transformed into a numerical variable, when assigning numbers (1 for SC and 2 for SS) and the correlations from a multivariate analysis could be obtained and interpreted. We believe that this section is important, due to correlations found for all factors and variables descripted in this manuscript. We could also see that the results from this table agree with the results from table 3 and we can have a better perspective of which variable were affected in higher or lower intensity by the main factors.      

 

Q27: Ln326-348: this entire section is based on a lot of assumptions.  Animals were not managed or fed the same or in a controlled environment.  Animal age could just as easily contribute to weight differences as differences in diet, which you have no idea what animals were fed.  At least you don’t share that information.  You didn’t quantify IMF so the discussion in this whole section is pretty far fetched for me.  I would delete or significantly revise based on what you actually know and what you actually tested and accounted for. 

A27: We partially agree on your point of view. We did not measure IMF, and just light conclusions can be obtained from the diet, since animals were not fed the same feed or in controlled conditions. However, we think that some of this relationships based exclusively on the analysis of the statistical data presented in this manuscript are important to develop our ideas about meat quality. We deleted the data that was based on general assumptions about IMF and we reformulated this paragraph to make our ideas more precise.

 

Q28: Ln350: I’m struggling to see how you would see metmyoglobin accumulation in a wet aging environmental, especially if only for 2 weeks.  Wet aging is a no/low O2 environment and O2 is required for oxidation and MMb accumulation. 

A28: This is a very fair point! We are also extremely curious as you are, to understand the mechanisms involved in this muscle discoloration. This is precisely the aim of our research project. We also believe this is quite an interesting topic, due to the reasons that you just mentioned. However, from our experience and from what our research partners have reported, color change can still happen 7 days after meat vacuum-packaging. Then, probably the mechanisms involved in this discoloration might have an important relationship with the remaining oxygen in the packaging. At low oxygen content (around 1%) the oxidation from myoglobin to metmyoglobin is increased in comparison to high oxygen atmosphere. This reaction leads to even a more stable product when myoglobin can react with oxygen to oxymyoglobin (King, 2023).

 

We also believe this could be a microbiological issue and we are currently researching if any bacteria present on the meat due to cross contamination may be responsible for it. However, we also believe that not a single factor, but a concatenation of factors may be linked to meat discoloration processes.     

 

àPD: In line 163, there was a small mistake. The samples were cooked until a core temperature of 72°C and not 75° C. We apologize for this error.

 

References:

Avery, P. J., & Masters, G. A. (1999). Advice on the Design and Analysis of Sensory Evaluation Experiments Using a Case-study with Cooked Pork. Journal of the Royal Statistical Society: Series D (The Statistician), 48(3), 349-359. doi:https://doi.org/10.1111/1467-9884.00193

Ellies-Oury, M.-P., Lorenzo, H., Denoyelle, C., Saracco, J., & Picard, B. (2019). An Original Methodology for the Selection of Biomarkers of Tenderness in Five Different Muscles. Foods, 8(6), 206. Retrieved from https://www.mdpi.com/2304-8158/8/6/206

Hernández, B., Sáenz Gamasa, C., Diñeiro Rubial, J. M., & Alberdi Odriozola, C. (2019). CIELAB color paths during meat shelf life. Meat Sci., 157, 107889. doi:https://doi.org/10.1016/j.meatsci.2019.107889

King, D., Melvin C. Hunt Shai Barbut James R. Claus Darren P. Cornforth Poulson Joseph Yuan H. Brad Kim Gunilla Lindahl Richard A. Mancini Mahesh N. Nair Kjell J. Merok Andy Milkowski Anand Mohan Fred Pohlman Ranjith Ramanathan Christopher R. Raines Mark Seyfert Oddvin Sørheim Surendranath P. Suman Mellissa Weber. (2023). American Meat Science Association Guidelines for Meat Color Measurement. Meat and Muscle Biology, 6(4). Retrieved from https://www.iastatedigitalpress.com/mmb/article/id/12473/

Poveda-Arteaga, A., Krell, J., Gibis, M., Heinz, V., Terjung, N., & Tomasevic, I. (2023). Intrinsic and Extrinsic Factors Affecting the Color of Fresh Beef Meat&mdash;Comprehensive Review. Applied Sciences, 13(7), 4382. Retrieved from https://www.mdpi.com/2076-3417/13/7/4382

Tomasevic, I., Djekic, I., Font-i-Furnols, M., Terjung, N., & Lorenzo, J. M. (2021). Recent advances in meat color research. Current Opinion in Food Science, 41, 81-87. doi:https://doi.org/10.1016/j.cofs.2021.02.012

Tomasevic, I., Tomovic, V., Ikonić, P., Lorenzo, J. M., Barba, F., Djekic, I., Nastasijevic, I., Stajić, S., & Zivkovic, D. (2019). Evaluation of poultry meat colour using computer vision system and colourimeter: Is there a difference? Br. Food J., 121, 1078-1087. doi:10.1108/BFJ-06-2018-0376

 

 

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

I am pleased with how the authors have handled the requests for changes. I believe the paper is now eligible for publication in Applied Science.

Author Response

Thank you so much for your comments. We are very glad to hear that 

Reviewer 2 Report

Comments and Suggestions for Authors

Q1: In terms of structure, as it is, it seems that lairage, mixing of unfamiliar animals and slaughtering temperature were treatments studied by the authors, as are transport distance (TD), animal weight (AW), and muscle position (MP). However, if that was the case, no data are presented to support that idea. In fact, about the former factors, the authors simply indicate the conditions verified during the study, without showing any proper analysis. The authors, themselves start the abstract saying “This study investigated the potential effects of transport distance, animal weight and muscle position on meat quality, under commercial conditions across four slaughtering weeks during the summer months (May to September). Data on transport, lairage and slaughtering were collected” So, data on transport, lairage and slaughtering should be presented in “Materials and Methods” and not in “Results and Discussion”. Also, it is strange to call Table 6, before calling (and presenting) Tables 3, 4 and 5. It is questionable, but if the authors want to start the “Results and Discussion” with an overall analysis (in line with the way they start the “Conclusions”), maybe it would be better to eliminate section “3.9. Multivariate test analysis of factor combination on meat quality traits” and include its contents in section 3.1, renumbering Table 6 to Table 3 (and correcting the number of Tables 3, 4 and 5). Also, the title of section 3.1 could be changed to something like “3.1. Combined effects of transport distance (TD), animal weight (AW), and muscle position (MP) on meat quality traits" since sections 3.3, 3.4 and 3.6 deal with the effect of each factor separately...

A1: Dear reviewer, thank you very much for your suggestions. We moved section 3.9 to 3.1 and numerated all tables as you recommended.

The data of transport, slaughtering and lairage were indeed presented in Tables 1 and 2 in the materials and methods section. The discussion of these two factors in the results and discussion section was interpreted according to the information obtained from the statistical model and references were cited to provide a background for this purpose.

Q1: The authors give indeed data about the actual conditions verified in their study in terms of lairage, mixing of unfamiliar animals and slaughtering temperature, but just point out in the “Results and discussion” section that, according to the literature, such factors were within intervals that would not cause stress to the animals. So, they didn’t really analyze these factors in their study – just contextualized their values using data from other studies. This means that saying “Therefore, the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed.” (lines 82-83) is misleading. This also applies to “animal transport”, since the only factor of animal transport analyzed was “transport distance”.  The authors are clear saying “This study investigated the potential effects of transport distance, animal weight and muscle position on meat quality (…)” (lines 12-13). They are also clear saying “Data on transport, lairage and slaughtering were collected” (line 14). It is not so clear that they just compared the conditions of their experiment with data on lairage, mixing of unfamiliar animals and slaughtering temperature collected from the literature. All the information on lairage, mixing of unfamiliar animals and slaughtering temperature is important but would be better placed in a section describing the conditions of the study, usually “Materials and Methods”, than in “Results and Discussion”. Eventually such information could be given at the beginning “Results and Discussio”, as a kind of introduction, but certainly not deserving especial sections or sub-sections.

 

Q2: LINE 18 – Is it really necessary to put ΔE here?

A2: It has been stated that color is one of the most important parameters in meat quality, from a consumer’s perspective (Tomasevic et al., 2021). The importance of ∆E in the present study is crucial, since the human eye can process color as a tridimensional stimuli (Hernández et al., 2019). Therefore, these values are indeed very important to compare the total color difference between shoulder clod and silverside and to determine if consumers can visually predict which muscles belong to each muscle position. However, we agree with you that this content cut the flow of the ideas in the abstract. Therefore, we changed it at the end of the section to increase the clarity of the ideas. We also added a couple of sentences to enhance the context the text.

Q2: No question at all about the importance of color. My point was just a minor one – the use of the acronym “∆E”, since it is not used anymore in the abstract. Why not just “color difference” instead of “color difference (∆E)”? Anyway, with the changes introduced, the abstract now “flows” better.

Q3: LINES 78-79 – The authors mention “For this reason, the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed.”. Throughout the paper there are several references to temperature, and temperature interval, but how was the temperature effect analysed?

A3: Thank you for pointing out this issue. In this sentence in specific we refer to meat temperature because pH post-mortem decline is closely related to this parameter. In some other parts of the text, the word “temperature” is referred to air temperature during slaughtering, which can also have a deep effect on meat quality (Poveda-Arteaga et al., 2023).  

Q3: Okay, but a question remains -  saying “(..) the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed is misleading. As I understand, in the present study, data was just “recorded”, not “analysed”…

(…)

Q5: LINES 207-209 – In Table 3, although it is a common notation, the meaning of different (or not) superscripts should be explained...

A5: We agree on that. We changed this as indicated.

Q5: Just a small note - although it is obvious, in general it is used “within lines”… So, in line 244 it would be “Superscripts not sharing a common letter, within lines, were significantly different (P<0.05).”

Q6: LINES 286-287 – The authors say “The meat from animals of short TD also exhibited a softer meat texture than the other groups, as determined through instrumental measurements and sensory evaluation.” What data support this? were the differences between TD treatments significant?

A6: This is correct. This information was taken from table 5. We also incorporated the superscripts for TD to differentiate between significant and non-significant values.

Q6: Am I missing something or in Table 6 there were no significant differences, for WBSF nor for hardness, concerning TD, although in Table 5 there is indeed a significant (although small) negative correlation with TD, both for WBSF and hardness?

Q7: LINES 303-305 – In Table 5, What do "Smell %", "Taste %" and "Texture %" mean? Since the authors make a parallel between WBSF values and sensory tenderness values (e.g., lines 355-366), how do they compare WBSF (%) with Texture (%)?

A7: Meat taste, smell and hardness were measured using the punctuation of a 1 to 100 scale, being in all cases 1 for “not perceptible” and 100 for “intense”. This description was added in the text and in the table explanation. Also, the percentage symbol was erased, since this measurements considered a scale and not a percentage per se.

Q7: Okay for text (lines 169-171), but still lacking for Tables 4, 5 and 6, and the percentage symbol is still in Tables 4, 5 and 6…

From our point of view, the sensory evaluation of meat texture is proportionally related with the instrumental measurement of shear force. That is why we used this correlation to describe meat quality.  

Q8: LINES 321-323 – The authors say “Short processing times were reflected in superior meat quality, due to pH values lower than 5.8 in all treatment combinations.“ . What data led the authors to conclude that the lower than 5.8 pH values observed were due to short processing?

A8: It has been mentioned in the literature that when beef attains pH values lower than around 5.8, then it is considered to have improved texture and color than meat from higher pH values, which falls into the DFD status (Lomiwes et al., 2013; McKeith et al., 2016; Page et al., 2001). We concluded that these relatively normal pH₄₈ values from all treatments are linked to fast processing times that we saw in our experiments (Table 2), also because we did not find any important color defects in any of our samples.

Q8: The authors seem to rely just on data from previous studies… How can they take such conclusion from the present study. Did they actually study the effect of short processing?  

 (…)

Q11: LINES 341-343 – The authors say “Increased tenderness could also be found from high AW combinations from both WBSF measurements and from the consumer’s point of view”. What data support this? Were the differences between TD treatments significant?

A11: This conclusion is supported by table 5. As in the previous question, we can only imply these results as a trend and significance does not apply in this example, because this factor was only divided into two groups.

Q11: My question was obviously about AW and not TD, but I think the authors understood it. Still I apologize. Anyway, it must be clearly stated that it was just a trend.

 

Q12: LINES 363-364 – The authors say “The SC muscles showed a higher ultimate pH than the SS, which subsequently resulted in improved color readings (L*, a* and b*) than SS.”. The authors had already "touched" this somehow (lines 227-230 and 233-235) when mentioning the correlations. Anyway, put this way, it seems that MP had a significant effect on all these parameters...

A 12: This information is correct. This comparison was made when writing about the Pearson correlations between main factors and variables and indicated that there was a negative significant correlation between muscle position and pH₄₈. However, we cannot say that this was a significant correlation, also because the factor MP was designed to have only in two levels.

Values from Pearson and Spearman correlations were very similar. However, just to be in the safe side, we also consider that when calculating the Spearman nonparametric coefficients, this test is independent from normality infractions that may have occurred in the model.

Q12: So, it must be clear there where just trends, concerning MP effects.

Q 13: LINES 364-366 – The authors say “Interestingly, higher pH values in the SC were not correlated neither with WBSF values or with sensory tenderness values and showed that SC were rated as more tender than SS.”. Again - were all these differences significant? What data support such conclusion? Another question - the samples tested were from fried meatballs made with minced meat obtained from SC and SS and not actual samples of meat slices from SC and SS. Right? While that is mentioned in "Introduction" and "Materials and Methods" the way it is presented here may induce the reader in error...  finally - along the paper the authors mix "texture" and "tenderness", and make a parallel between "WBSF values" and "sensory tenderness values"...  While  many papers don't distinguish between "texture" and "tenderness" the fact is that texture of meat refers to firmness (toughness or degree of tenderness), cohesiveness and juiciness. I would suggest the authors clearly state what they analysed (texture or tenderness), avoiding to mention sometimes "tenderness" and other times "texture", throughout the paper.

A 13 :Thank you bringing this matter into our attention. It has been previously published that problems associated with high pH meat can also lead to harder texture (Ponnampalam et al., 2017). Therefore, we expected to obtain tougher meat from SC, because it had higher pH values than SS as shown in Table 5. However, our data suggested that WBSF values were higher in SS and not in SC. From a sensory perspective, same correlations were obtained and showed that SS was tougher than SC, despite of having lower pH. No post-hoc test is available, since there are only two means, and the information can only be compared, but no significance level can be calculated.    

Q 13: Okay. However, When the authors say in the revised version “Interestingly, higher pH values in the SC were not correlated neither with WBSF values nor with sensory hardness values and showed that SC were rated as softer muscles (lines 416-418)” it seems that higher pH values showed that SC were rated as softer muscles” despite the correlation or not with WBSF and sensory hardness… Is this what the authors really want to say?  

 For your second question, the answer is yes, this is right. Texture analysis was in both cases made from minced meat and not from the actual steaks. The reason why we designed the experiment in this way is because we were aiming to have more uniform samples. If single steaks would have been taken for every measurement, this would have taken much more time and probably less reliable results could have been obtained. Also, different steaks had also different fat contents, which would have affected the clarity of the results. Therefore, steaks were trimmed from the fat before mincing the meat.   

I understand. However, it could be better if the authors stated, for instance in line 194, stand for muscles for minced meat from muscles “so and so” and throughout the text only use the acronyms, without speaking of muscles.

And for the third point, we also agree with your opinion. Texture is a wider quality attribute in meat. However, it is difficult to just attach to only one of the two terminologies since the development of the ideas sometimes need one or the other. We tried to keep it as straight forward as possible to prevent the indiscriminate use of those two words.

Q 14: LINES 368-370 – The authors say “Taste and smell values were also higher for SC than SS and reaffirmed that in this case, these sensory traits were not related to pH development in meat.”. Again - were these differences significant? What data support such conclusion? What is the scale of these "taste and smaell values"

A 14: Taste, smell and hardness were measured using the punctuation from a 1 to 100 scale, being in all cases 1 for “not perceptible” and 100 for “intense”. The idea of this sentence was to emphasize that even if higher values for smell and taste were expected from SS, which is an indicator for superior meat quality, exactly the contrary occurred. 

Q 14: I understand it is monotonous but, once more, there were just trends. And this must be clear.

Q 15: LINES 381-383 – The authors say “In the present study, cattle were carefully transported to the abattoir in small lots of familiar animals that indicated lower pre-mortem stress and higher meat quality attributes.” What data "indicated" lower pre-mortem stress and higher meat quality attributes?

A 15: From our point of view, lower pre-mortem stress was reflected in overall low pH values of all meat pieces studied. The data presented in Table 5 indicated that any of the treatments had pH values higher than 5.8, which was determinant in enhancing meat quality (Poveda-Arteaga et al., 2023; Węglarz, 2010). Other studies indicated that higher pH values can be obtained from cattle that had longer transportation times before arriving to the abattoir (Romero et al., 2013). Also, the fact that ante-mortem stress increases when cattle are mixed with unfamiliar animals on their way to the slaughterhouse (Mounier et al., 2006).

In our case, animal stress was reduced by transporting only familiar animals, which were separated in the pins before slaughtering.

Q15: The fact that “cattle were carefully transported to the abattoir in small lots of familiar animals” is an indication of “lower pre-mortem stress and higher meat quality attributes.” or a potential reason for “lower pre-mortem stress and higher meat quality attributes.”? I would agree more if the authors stated, as in their answer, that lower pre-mortem stress was reflected in overall low pH values of all meat pieces studied and that was a possible consequence of  having been carefully transported to the abattoir in small lots of familiar animals.

Q 16: LINES 384-385 – The authors say “stress-related problems did not seriously affect the physiological status of the young bulls.”. What data allow the authors conclude this?

A 16: As previously described, this information was concluded based on observed pH values of all treatments collected in this experiment ranging between 5.47 to 5.78.

Q16: Okay. Then why not state precisely that, relating to the question 15?

(…)

Q 20: LINES 424-427 – The authors say “Mild temperatures during the slaughtering days between May and September and short lairage times (<2 hours) did not importantly affect the physiological status of the young bulls and might have also contributed to improve the meat quality parameters.”. As mentioned before - where is the data supporting this?

A 20: This data can be obtained from Table 2, which has data concerning air temperature and lairage times for all animals from this study. We added this reference in the text as suggested.

Q 20: I don’t think the authors can take a conclusion about a factor that was not tested in their study. It is just a speculation using data obtained in other studies..

Q 21: LINES 430-431 – The authors say “higher pH48 values were obtained from the shoulder clod, which resulted in lower L*, a* and b* readings.”. Why do the authors conclude that lower L*, a* and b* readings were consequence of higher pH48 values?

A 21: Several reviews in the field suggested that high pH values are directly related to dark meat colors, a defect which is better known as DFD meat (Neethling et al., 2017; Poveda-Arteaga et al., 2023; Ruedt et al., 2023; Tomasevic et al., 2021). A combination of factors may increase the probability to find these problems in beef and animal stress is recognized as one of the most important reasons why this issue may occur. Therefore, we concluded that as pH values get closer to 5.6, then the color coordinates of the beef pieces in our experiment will also increase as shown in Table 5.  

Q21: Again, the authors are comparing a trend observed in their study with data from other studies. That makes sense in the discussion. However, I don’t think it can be put as such clear conclusion of the present study.

Q22: LINES 434-437 – The authors say “Future investigations should consider other year seasons and more extreme temperatures to find if variable weather conditions exert a more drastic influence on the meat quality of specific cuts such as shoulder-clod and silverside.”. Please see above note about the temperature effect.

A 22: We speculate that this may be an interesting topic for future investigations. Because animals were not exposed to extreme low or high temperatures in our study, we assume that this should be investigated over a longer time span.

Q21: I agree it is an interesting topic. It just happens that the possible effect of ambient temperature wasn’t something analyzed in the present study.

 

Author Response

Response to reviewer 2

 

 

Q1: In terms of structure, as it is, it seems that lairage, mixing of unfamiliar animals and slaughtering temperature were treatments studied by the authors, as are transport distance (TD), animal weight (AW), and muscle position (MP). However, if that was the case, no data are presented to support that idea. In fact, about the former factors, the authors simply indicate the conditions verified during the study, without showing any proper analysis. The authors, themselves start the abstract saying “This study investigated the potential effects of transport distance, animal weight and muscle position on meat quality, under commercial conditions across four slaughtering weeks during the summer months (May to September). Data on transport, lairage and slaughtering were collected” So, data on transport, lairage and slaughtering should be presented in “Materials and Methods” and not in “Results and Discussion”. Also, it is strange to call Table 6, before calling (and presenting) Tables 3, 4 and 5. It is questionable, but if the authors want to start the “Results and Discussion” with an overall analysis (in line with the way they start the “Conclusions”), maybe it would be better to eliminate section “3.9. Multivariate test analysis of factor combination on meat quality traits” and include its contents in section 3.1, renumbering Table 6 to Table 3 (and correcting the number of Tables 3, 4 and 5). Also, the title of section 3.1 could be changed to something like “3.1. Combined effects of transport distance (TD), animal weight (AW), and muscle position (MP) on meat quality traits" since sections 3.3, 3.4 and 3.6 deal with the effect of each factor separately...

A1: Dear reviewer, thank you very much for your suggestions. We moved section 3.9 to 3.1 and numerated all tables as you recommended.

The data of transport, slaughtering and lairage were indeed presented in Tables 1 and 2 in the materials and methods section. The discussion of these two factors in the results and discussion section was interpreted according to the information obtained from the statistical model and references were cited to provide a background for this purpose.

Q1.1: The authors give indeed data about the actual conditions verified in their study in terms of lairage, mixing of unfamiliar animals and slaughtering temperature, but just point out in the “Results and discussion” section that, according to the literature, such factors were within intervals that would not cause stress to the animals. So, they didn’t really analyze these factors in their study – just contextualized their values using data from other studies. This means that saying “Therefore, the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed.” (lines 82-83) is misleading. This also applies to “animal transport”, since the only factor of animal transport analyzed was “transport distance”.  The authors are clear saying “This study investigated the potential effects of transport distance, animal weight and muscle position on meat quality (…)” (lines 12-13). They are also clear saying “Data on transport, lairage and slaughtering were collected” (line 14). It is not so clear that they just compared the conditions of their experiment with data on lairage, mixing of unfamiliar animals and slaughtering temperature collected from the literature. All the information on lairage, mixing of unfamiliar animals and slaughtering temperature is important but would be better placed in a section describing the conditions of the study, usually “Materials and Methods”, than in “Results and Discussion”. Eventually such information could be given at the beginning “Results and Discussion”, as a kind of introduction, but certainly not deserving especial sections or sub-sections.

A1.1: We agree with you that the data from lairage, mixing unfamiliar animals and slaughtering temperature were exclusively collected and compared with the results of previous research. We tried to put it in this context. However, we consider that this information is part of results and conclusions. We rewrote the whole three sections and made an introductory paragraph in the results and discussion section with the data collected from these parameters. The conclusions obtained from these parameters were added accordingly in the conclusion section. We also changed the information from lines 79 and 80.  

Q2: LINE 18 – Is it really necessary to put ΔE here?

A2: It has been stated that color is one of the most important parameters in meat quality, from a consumer’s perspective (Tomasevic et al., 2021). The importance of ∆E in the present study is crucial, since the human eye can process color as a tridimensional stimuli (Hernández et al., 2019). Therefore, these values are indeed very important to compare the total color difference between shoulder clod and silverside and to determine if consumers can visually predict which muscles belong to each muscle position. However, we agree with you that this content cut the flow of the ideas in the abstract. Therefore, we changed it at the end of the section to increase the clarity of the ideas. We also added a couple of sentences to enhance the context the text.

Q2.1: No question at all about the importance of color. My point was just a minor one – the use of the acronym “∆E”, since it is not used anymore in the abstract. Why not just “color difference” instead of “color difference (∆E)”? Anyway, with the changes introduced, the abstract now “flows” better.

A2.1: We appreciate your comment. We erased the acronym for color difference as suggested.

Q3: LINES 78-79 – The authors mention “For this reason, the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed.”. Throughout the paper there are several references to temperature, and temperature interval, but how was the temperature effect analysed?

A3: Thank you for pointing out this issue. In this sentence in specific we refer to meat temperature because pH post-mortem decline is closely related to this parameter. In some other parts of the text, the word “temperature” is referred to air temperature during slaughtering, which can also have a deep effect on meat quality (Poveda-Arteaga et al., 2023).  

Q3.1: Okay, but a question remains -  saying “(..) the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed is misleading. As I understand, in the present study, data was just “recorded”, not “analysed”…

A3.1: Thank for clarifying this issue. You are right. We changed the meaning of the sentence as suggested.

Q5: LINES 207-209 – In Table 3, although it is a common notation, the meaning of different (or not) superscripts should be explained…

A5: We agree on that. We changed this as indicated.

Q5: Just a small note – although it is obvious, in general it is used “within lines”… So, in line 244 it would be “Superscripts not sharing a common letter, within lines, were significantly different (P<0.05).”

A5.1: Thank you for your comment. We changed this as proposed.

Q6: LINES 286-287 – The authors say “The meat from animals of short TD also exhibited a softer meat texture than the other groups, as determined through instrumental measurements and sensory evaluation.” What data support this? were the differences between TD treatments significant?

A6: This is correct. This information was taken from table 5. We also incorporated the superscripts for TD to differentiate between significant and non-significant values.

Q6: Am I missing something or in Table 6 there were no significant differences, for WBSF nor for hardness, concerning TD, although in Table 5 there is indeed a significant (although small) negative correlation with TD, both for WBSF and hardness?

A6.1: In this case, we think you are wrong. For both Spearman non-parametric correlations and for the MANOVA model, there were no significance for WBSF or hardness. Probably, in this case you are confusing the Rho scores with significance value.

Q7: LINES 303-305 – In Table 5, What do "Smell %", "Taste %" and "Texture %" mean? Since the authors make a parallel between WBSF values and sensory tenderness values (e.g., lines 355-366), how do they compare WBSF (%) with Texture (%)?

A7: Meat taste, smell and hardness were measured using the punctuation of a 1 to 100 scale, being in all cases 1 for “not perceptible” and 100 for “intense”. This description was added in the text and in the table explanation. Also, the percentage symbol was erased, since this measurements considered a scale and not a percentage per se.

Q7: Okay for text (lines 169-171), but still lacking for Tables 4, 5 and 6, and the percentage symbol is still in Tables 4, 5 and 6…

A7.1: Sorry about this misunderstanding. It is possible that we did not erase them in your corrections, but indeed, we erased the percentage symbol in all tables at the final version.  

From our point of view, the sensory evaluation of meat texture is proportionally related with the instrumental measurement of shear force. That is why we used this correlation to describe meat quality.  

Q8: LINES 321-323 – The authors say “Short processing times were reflected in superior meat quality, due to pH values lower than 5.8 in all treatment combinations.“ . What data led the authors to conclude that the lower than 5.8 pH values observed were due to short processing?

A8: It has been mentioned in the literature that when beef attains pH values lower than around 5.8, then it is considered to have improved texture and color than meat from higher pH values, which falls into the DFD status (Lomiwes et al., 2013; McKeith et al., 2016; Page et al., 2001). We concluded that these relatively normal pH₄₈ values from all treatments are linked to fast processing times that we saw in our experiments (Table 2), also because we did not find any important color defects in any of our samples.

Q8: The authors seem to rely just on data from previous studies… How can they take such conclusion from the present study. Did they actually study the effect of short processing?  

A8.1: We assume that low pH values resulted from a combination of factors during transport, lairage and slaughtering. None of our treatments exceeded pH=5.8, which is the reason why we imply that fast processing times (less than 2 hours of lairage of our animals) was related with improved meat quality. We changed this in the final version of the manuscript, and we included it in the conclusions as follows:

Our study suggested that reduced animal stress did not seriously affect the physical or physiological status of the young bulls at any point during transport, lairage and slaughtering. Several factors contributed for these results: firstly, our animals had to wait in the lairage pens for times lesser than two hours until being slaughtered; also, the cattle in the present study were carefully transported to the abattoir in small lots of familiar animals and mixing of unfamiliar animals was prevented during lairage; and finally, the average daily temperatures during the slaughtering days were between 18.75 and 10°C, with temperature fluctuations not higher 8°C in all cases. These conditions resulted in pH values lower than 5.8 in all treatment combinations and a* threshold values higher than the limit recommended of 14.5, which implied that consumers could accept the meat from all treatments.

Q11: LINES 341-343 – The authors say “Increased tenderness could also be found from high AW combinations from both WBSF measurements and from the consumer’s point of view”. What data support this? Were the differences between TD treatments significant?

A11: This conclusion is supported by table 5. As in the previous question, we can only imply these results as a trend and significance does not apply in this example, because this factor was only divided into two groups.

Q11: My question was obviously about AW and not TD, but I think the authors understood it. Still I apologize. Anyway, it must be clearly stated that it was just a trend.

A11.1: No problem. We stated in the final version that heavier animals tended to achieve lower pH values.

Q12: LINES 363-364 – The authors say “The SC muscles showed a higher ultimate pH than the SS, which subsequently resulted in improved color readings (L*, a* and b*) than SS.”. The authors had already "touched" this somehow (lines 227-230 and 233-235) when mentioning the correlations. Anyway, put this way, it seems that MP had a significant effect on all these parameters...

A 12: This information is correct. This comparison was made when writing about the Pearson correlations between main factors and variables and indicated that there was a negative significant correlation between muscle position and pH₄₈. However, we cannot say that this was a significant correlation, also because the factor MP was designed to have only in two levels.

Values from Pearson and Spearman correlations were very similar. However, just to be in the safe side, we also consider that when calculating the Spearman nonparametric coefficients, this test is independent from normality infractions that may have occurred in the model.

Q12: So, it must be clear there where just trends, concerning MP effects.

A112.1: Thank you for your comment. We stated in the final version that SC muscles tended to achieve higher pH values.

Q 13: LINES 364-366 – The authors say “Interestingly, higher pH values in the SC were not correlated neither with WBSF values or with sensory tenderness values and showed that SC were rated as more tender than SS.”. Again - were all these differences significant? What data support such conclusion? Another question - the samples tested were from fried meatballs made with minced meat obtained from SC and SS and not actual samples of meat slices from SC and SS. Right? While that is mentioned in "Introduction" and "Materials and Methods" the way it is presented here may induce the reader in error...  finally - along the paper the authors mix "texture" and "tenderness", and make a parallel between "WBSF values" and "sensory tenderness values"...  While  many papers don't distinguish between "texture" and "tenderness" the fact is that texture of meat refers to firmness (toughness or degree of tenderness), cohesiveness and juiciness. I would suggest the authors clearly state what they analysed (texture or tenderness), avoiding to mention sometimes "tenderness" and other times "texture", throughout the paper.

A 13 :Thank you bringing this matter into our attention. It has been previously published that problems associated with high pH meat can also lead to harder texture (Ponnampalam et al., 2017). Therefore, we expected to obtain tougher meat from SC, because it had higher pH values than SS as shown in Table 5. However, our data suggested that WBSF values were higher in SS and not in SC. From a sensory perspective, same correlations were obtained and showed that SS was tougher than SC, despite of having lower pH. No post-hoc test is available, since there are only two means, and the information can only be compared, but no significance level can be calculated.    

Q 13: Okay. However, When the authors say in the revised version “Interestingly, higher pH values in the SC were not correlated neither with WBSF values nor with sensory hardness values and showed that SC were rated as softer muscles (lines 416-418)” it seems that higher pH values showed that SC were rated as softer muscles” despite the correlation or not with WBSF and sensory hardness… Is this what the authors really want to say?

Q13.1.1: This is correct. We tried to highlight that despite of higher pH values in SC, those muscles were rated as softer using both instrumental and sensorial texture analyses. As mentioned before, this relation is contradictory, since it has been informed that higher pH values lead to a harder texture. We included this information in the manuscript to emphasize our point of view regarding this issue.

For your second question, the answer is yes, this is right. Texture analysis was in both cases made from minced meat and not from the actual steaks. The reason why we designed the experiment in this way is because we were aiming to have more uniform samples. If single steaks would have been taken for every measurement, this would have taken much more time and probably less reliable results could have been obtained. Also, different steaks had also different fat contents, which would have affected the clarity of the results. Therefore, steaks were trimmed from the fat before mincing the meat.   

I understand. However, it could be better if the authors stated, for instance in line 194, stand for muscles for minced meat from muscles “so and so” and throughout the text only use the acronyms, without speaking of muscles.

Q13.1.2: Thanks for your comment. However, we believe that using acronyms for this reason might, in some cases, lower the clarity of our ideas. Therefore, we did not accept this suggestion.

And for the third point, we also agree with your opinion. Texture is a wider quality attribute in meat. However, it is difficult to just attach to only one of the two terminologies since the development of the ideas sometimes need one or the other. We tried to keep it as straight forward as possible to prevent the indiscriminate use of those two words.

Q 14: LINES 368-370 – The authors say “Taste and smell values were also higher for SC than SS and reaffirmed that in this case, these sensory traits were not related to pH development in meat.”. Again - were these differences significant? What data support such conclusion? What is the scale of these "taste and smaell values"

A 14: Taste, smell and hardness were measured using the punctuation from a 1 to 100 scale, being in all cases 1 for “not perceptible” and 100 for “intense”. The idea of this sentence was to emphasize that even if higher values for smell and taste were expected from SS, which is an indicator for superior meat quality, exactly the contrary occurred. 

Q 14: I understand it is monotonous but, once more, there were just trends. And this must be clear.

Q14.1: Thank you for your suggestion. We changed this.

Q 15: LINES 381-383 – The authors say “In the present study, cattle were carefully transported to the abattoir in small lots of familiar animals that indicated lower pre-mortem stress and higher meat quality attributes.” What data "indicated" lower pre-mortem stress and higher meat quality attributes?

A 15: From our point of view, lower pre-mortem stress was reflected in overall low pH values of all meat pieces studied. The data presented in Table 5 indicated that any of the treatments had pH values higher than 5.8, which was determinant in enhancing meat quality (Poveda-Arteaga et al., 2023; Węglarz, 2010). Other studies indicated that higher pH values can be obtained from cattle that had longer transportation times before arriving to the abattoir (Romero et al., 2013). Also, the fact that ante-mortem stress increases when cattle are mixed with unfamiliar animals on their way to the slaughterhouse (Mounier et al., 2006).

In our case, animal stress was reduced by transporting only familiar animals, which were separated in the pins before slaughtering.

Q15: The fact that “cattle were carefully transported to the abattoir in small lots of familiar animals” is an indication of “lower pre-mortem stress and higher meat quality attributes.” or a potential reason for “lower pre-mortem stress and higher meat quality attributes.”? I would agree more if the authors stated, as in their answer, that lower pre-mortem stress was reflected in overall low pH values of all meat pieces studied and that was a possible consequence of  having been carefully transported to the abattoir in small lots of familiar animals.

Q15.1: Thank you for your suggestion. You are right. We changed this in the conclusion section of the final version and we included the word potentially to focus our ideas in this direction.

Q 16: LINES 384-385 – The authors say “stress-related problems did not seriously affect the physiological status of the young bulls.”. What data allow the authors conclude this?

A 16: As previously described, this information was concluded based on observed pH values of all treatments collected in this experiment ranging between 5.47 to 5.78.

Q16: Okay. Then why not state precisely that, relating to the question 15?

Q16.1: We changed this in the conclusion as well.

Q 20: LINES 424-427 – The authors say “Mild temperatures during the slaughtering days between May and September and short lairage times (<2 hours) did not importantly affect the physiological status of the young bulls and might have also contributed to improve the meat quality parameters.”. As mentioned before - where is the data supporting this?

A 20: This data can be obtained from Table 2, which has data concerning air temperature and lairage times for all animals from this study. We added this reference in the text as suggested.

Q 20: I don’t think the authors can take a conclusion about a factor that was not tested in their study. It is just a speculation using data obtained in other studies.

Q20.1: Thanks for your comment. You are right in this affirmation. Meat quality traits do not depend on a single factor, but from a combination of factors. That is why in the conclusion we included this information and we merely speculated that a factor combination might have acted for achieving improved meat quality.

Q 21: LINES 430-431 – The authors say “higher pH48 values were obtained from the shoulder clod, which resulted in lower L*, a* and b* readings.”. Why do the authors conclude that lower L*, a* and b* readings were consequence of higher pH48 values?

A 21: Several reviews in the field suggested that high pH values are directly related to dark meat colors, a defect which is better known as DFD meat (Neethling et al., 2017; Poveda-Arteaga et al., 2023; Ruedt et al., 2023; Tomasevic et al., 2021). A combination of factors may increase the probability to find these problems in beef and animal stress is recognized as one of the most important reasons why this issue may occur. Therefore, we concluded that as pH values get closer to 5.6, then the color coordinates of the beef pieces in our experiment will also increase as shown in Table 5.  

Q21: Again, the authors are comparing a trend observed in their study with data from other studies. That makes sense in the discussion. However, I don’t think it can be put as such clear conclusion of the present study.

Q21.1: We appreciate your comment. However, we do not share your position. We believe that this is one of the important conclusions that we got from the MANOVA model, and this is why we did not erase this information from the final version.

Q22: LINES 434-437 – The authors say “Future investigations should consider other year seasons and more extreme temperatures to find if variable weather conditions exert a more drastic influence on the meat quality of specific cuts such as shoulder-clod and silverside.”. Please see above note about the temperature effect.

A 22: We speculate that this may be an interesting topic for future investigations. Because animals were not exposed to extreme low or high temperatures in our study, we assume that this should be investigated over a longer time span.

Q21: I agree it is an interesting topic. It just happens that the possible effect of ambient temperature wasn’t something analyzed in the present study.

Q21.1: Thank you for your comment. We agree with you. This information was erased from the conclusions.

Author Response File: Author Response.docx

Reviewer 3 Report

Comments and Suggestions for Authors

Q2: Ln14: specifically what data were collected? transport time and distance, lairage time, and what about slaughtering??

A2: All the pre-mortem parameters that we could collect from the slaughterhouse about transport, lairage and slaughtering conditions of the animals were condensed and presented in tables 1 and 2. With slaughtering we mean the weather conditions on those days, which is an important parameter that can influence animal stress and meat quality.  

Nothing was actually clarified in the abstract. 

 

Q4: Ln18, 42, and throughout: you’re not measuring a change, but simply a difference in color between two independent samples.  This is not a valid way to report color values in this study. 

A4: Thank you for clearing this out. We totally agree with you, and we checked again the whole manuscript to make sure that this information was presented as you suggested.

This is not delta E. Just refer to it as a difference between two muscles. Or better yet, simply report the values independently, since they are independent variables. 

 

Q12: Ln133-142: you do not mention anything about Delta E here.  I don’t see this as an appropriate way to report color in this study.  It was not a change in color.  You should report independent color values of the two muscles. 

A12: It has been stated that color is one of the most important parameters in meat quality, from a consumer’s perspective (Tomasevic et al., 2021). The importance of ∆E in the present study is crucial since the human eye can process color as a tridimensional sensation (Hernández et al., 2019). Therefore, these values are indeed very important to compare the total color difference between shoulder clod and silverside and to determine if consumers can visually predict which muscles belong to each muscle position. However, individual color values of every treatment were also presented in a L*, a* and b* format as shown in Table 3.  

Yes, color is important to consumers and often used to make purchase decisions.  Delta E in the present study does nothing to explain the tridimensional sensation you suggest it does.  Instead you should calculate hue, chroma, and saturation and report for each muscle. 

 

Q15: Ln163: please provide training protocols for panelists. How many sessions were conducted? 

A15: Thank you for these important comments. Before the initial sensory evaluation session, there was a brief introduction about the topic asked in the questionary and how meat traits should have been measured. The judges were selected from a group of workers at DIL that have previous experience in sensory analysis of meat samples. The questionary was self-explanatory. The sensory analysis was performed in four sessions in total, which is the number of working weeks that we got samples from the slaughterhouse.    

You didn’t answer the question of how the panelists were trained?  Please provide the reference for training protocols and the length of the training. If they weren’t trained then don’t say that they were.  Experience and training is not the same thing.  

 

A25:Thank you for your suggestions. We agree with you, and we included the information that you proposed in the table description and deleted the percentage symbol from all the parameters from sensory analysis in all tables.

 

About the ∆E values, it has been published that it is a sensitive indicator to measure myoglobin oxidation changes in beef muscles (Hernández et al., 2019). However, in this paper the purpose of ∆E calculation was to differentiate the color of the two muscle positions and not color difference inside the same muscle. Therefore, we consider that the differences measured between SS and SC were valid, because they considered only individual L*, a* and b* coordinates from those independent muscles from table 3. The L*, a* and b* values from SS and SC are also presented in the same table.

 

Also, it is important to consider that ∆E calculation was used previously, for example, to compare color measurements between computer vision system (CVS) and a Minolta colorimeter (Tomasevic et al., 2019). In another manuscript, ∆E was also used to compare the total color difference between two muscles of several publications using the L*, a* and b* values from individual muscles (Poveda-Arteaga et al., 2023).

You’ve given two examples of papers where delta E was used, but both are from this author group.  Just because you’ve managed to incorporate into previous papers, does not make it appropriate in this instance. 

 

 

Author Response

Response to reviewer 3

 

Q2: Ln14: specifically what data were collected? transport time and distance, lairage time, and what about slaughtering??

A2: All the pre-mortem parameters that we could collect from the slaughterhouse about transport, lairage and slaughtering conditions of the animals were condensed and presented in tables 1 and 2. With slaughtering we mean the weather conditions on those days, which is an important parameter that can influence animal stress and meat quality.  

Q 2.1: Nothing was actually clarified in the abstract. 

A2.1: We apologize for this misunderstanding. We referred to transport distance, lairage time and ambient temperature during slaughtering days. This information was added in the abstract to clarify how the information was collected.

Q4: Ln18, 42, and throughout: you’re not measuring a change, but simply a difference in color between two independent samples.  This is not a valid way to report color values in this study. 

A4: Thank you for clearing this out. We totally agree with you, and we checked again the whole manuscript to make sure that this information was presented as you suggested.

This is not delta E. Just refer to it as a difference between two muscles. Or better yet, simply report the values independently, since they are independent variables. 

A4.1: Thank you for the advice. We understand your idea now. We will report our results as you suggested.

Q12: Ln133-142: you do not mention anything about Delta E here.  I don’t see this as an appropriate way to report color in this study.  It was not a change in color.  You should report independent color values of the two muscles. 

A12: It has been stated that color is one of the most important parameters in meat quality, from a consumer’s perspective (Tomasevic et al., 2021). The importance of ∆E in the present study is crucial since the human eye can process color as a tridimensional sensation (Hernández et al., 2019). Therefore, these values are indeed very important to compare the total color difference between shoulder clod and silverside and to determine if consumers can visually predict which muscles belong to each muscle position. However, individual color values of every treatment were also presented in a L*, a* and b* format as shown in Table 3.  

Yes, color is important to consumers and often used to make purchase decisions.  Delta E in the present study does nothing to explain the tridimensional sensation you suggest it does.  Instead you should calculate hue, chroma, and saturation and report for each muscle. 

 A4.1: Thank you for your comment. We will report our results as you suggested. The values for chroma and hue were correspondingly added to tables 4, 5 and 6.

Q15: Ln163: please provide training protocols for panelists. How many sessions were conducted? 

A15: Thank you for these important comments. Before the initial sensory evaluation session, there was a brief introduction about the topic asked in the questionary and how meat traits should have been measured. The judges were selected from a group of workers at DIL that have previous experience in sensory analysis of meat samples. The questionary was self-explanatory. The sensory analysis was performed in four sessions in total, which is the number of working weeks that we got samples from the slaughterhouse.    

You didn’t answer the question of how the panelists were trained?  Please provide the reference for training protocols and the length of the training. If they weren’t trained then don’t say that they were.  Experience and training is not the same thing.  

A15.1: Thank you for your feedback. We think we did not do any special training procedure for this experiment. The judges were only informed about how to measure each parameter before the starting of the first session. We meant that they had experience in meat sensory, because the same judges were used for a similar project previously. We accepted you suggestion and we omitted the word “trained” in this manuscript.  

A25:Thank you for your suggestions. We agree with you, and we included the information that you proposed in the table description and deleted the percentage symbol from all the parameters from sensory analysis in all tables.

A25.1: We made sure that the percentage symbols were erased in all tables, since these parameters were calculated based on a 1 to 100 scale.

About the ∆E values, it has been published that it is a sensitive indicator to measure myoglobin oxidation changes in beef muscles (Hernández et al., 2019). However, in this paper the purpose of ∆E calculation was to differentiate the color of the two muscle positions and not color difference inside the same muscle. Therefore, we consider that the differences measured between SS and SC were valid, because they considered only individual L*, a* and b* coordinates from those independent muscles from table 3. The L*, a* and b* values from SS and SC are also presented in the same table.

Also, it is important to consider that ∆E calculation was used previously, for example, to compare color measurements between computer vision system (CVS) and a Minolta colorimeter (Tomasevic et al., 2019). In another manuscript, ∆E was also used to compare the total color difference between two muscles of several publications using the L*, a* and b* values from individual muscles (Poveda-Arteaga et al., 2023).

You’ve given two examples of papers where delta E was used, but both are from this author group.  Just because you’ve managed to incorporate into previous papers, does not make it appropriate in this instance. 

A25.1: Thank you for your comment. We did as suggested to enhance the quality of our manuscript.

Author Response File: Author Response.docx

Round 3

Reviewer 2 Report

Comments and Suggestions for Authors

Just some final remaks/suggestions:

Q3: LINES 78-79 – The authors mention “For this reason, the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed.”. (…)

(…)

Q3.1: Okay, but a question remains -  saying “(..) the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed is misleading. As I understand, in the present study, data was just “recorded”, not “analysed”…

A3.1: Thank for clarifying this issue. You are right. We changed the meaning of the sentence as suggested.

Q3.2: I received two different revised documents and so I am not sure what was the authors latest version, but would agree more with the one stating “Therefore, the information about transport distance, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were recorded.” (LINES 79-80). It is not a critical point.

Q6: LINES 286-287 – The authors say “The meat from animals of short TD also exhibited a softer meat texture than the other groups, as determined through instrumental measurements and sensory evaluation.” (…)

(…)

A6.1: In this case, we think you are wrong. For both Spearman non-parametric correlations and for the MANOVA model, there were no significance for WBSF or hardness. Probably, in this case you are confusing the Rho scores with significance value.

Q6.2: My mistake about the “significant (although small) negative correlation with TD”. Sorry about that (in fact my initial question had precisely to do with my doub about such significance, when reading the first version submitted). As I say above, I received two different revised documents, but would agree more with the one stating “No significant differences were found for meat hardness between different TD, both calculated by sensory analysis and WBSF” (LINES 369-370)

Q 13: LINES 364-366 – The authors say “Interestingly, higher pH values in the SC were not correlated neither with WBSF values or with sensory tenderness values and showed that SC were rated as more tender than SS.”. (…) the samples tested were from fried meatballs made with minced meat obtained from SC and SS and not actual samples of meat slices from SC and SS. (…).

(…)   

Q13.1.2: Thanks for your comment. However, we believe that using acronyms for this reason might, in some cases, lower the clarity of our ideas. Therefore, we did not accept this suggestion.

Q13.3: No criticism about not using individual steaks. I agree with the authors about the problem of using acronyms. My point was just about the importance of clarifying early in the text (and in table titles) that SC and SS refer, respectively to minced samples of shoulder clod and silverside, and not to actual shoulder clod and silverside muscles. Then it would be just a matter of using the acronyms. For other parameters, using muscle samples or minced muscle samples is not critical, but for WBSF and sensory tenderness can be.

Q 21: LINES 430-431 – The authors say “higher pH48 values were obtained from the shoulder clod, which resulted in lower L*, a* and b* readings.”. Why do the authors conclude that lower L*, a* and b* readings were consequence of higher pH48 values?

(…)

Q21.1: We appreciate your comment. However, we do not share your position. We believe that this is one of the important conclusions that we got from the MANOVA model, and this is why we did not erase this information from the final version.

Q21.2: My point is just about the strength of your conclusion and not so much mentioning it in “Conclusions”. If, instead of “which resulted” was “which tended to cause” would be less controversial.

Author Response

Dear reviewer

Thank you for your comments. We are confident that these final revisions will comply to your requests.

Q3: LINES 78-79 – The authors mention “For this reason, the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed.”. (…)

(…)

Q3.1: Okay, but a question remains -  saying “(..) the records of animal transport, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were measured and analyzed is misleading. As I understand, in the present study, data was just “recorded”, not “analysed”…

Q3.2: I received two different revised documents and so I am not sure what was the authors latest version, but would agree more with the one stating “Therefore, the information about transport distance, and the physico-chemical parameters of fresh meat such as pH, temperature, and color were recorded.” (LINES 79-80). It is not a critical point.

A1: Thank you for this comment. We changed it accordingly in the introduction as suggested.

Q6: LINES 286-287 – The authors say “The meat from animals of short TD also exhibited a softer meat texture than the other groups, as determined through instrumental measurements and sensory evaluation.” (…)

(…)

A6.1: In this case, we think you are wrong. For both Spearman non-parametric correlations and for the MANOVA model, there were no significance for WBSF or hardness. Probably, in this case you are confusing the Rho scores with significance value.

Q6.2: My mistake about the “significant (although small) negative correlation with TD”. Sorry about that (in fact my initial question had precisely to do with my doubt about such significance, when reading the first version submitted). As I say above, I received two different revised documents, but would agree more with the one stating “No significant differences were found for meat hardness between different TD, both calculated by sensory analysis and WBSF” (LINES 369-370)

A2: Thank you for your correction. We agree with your point of view and we will leave the conclusion of this section as “No significant differences were found for meat hardness between different TD, both calculated by sensory analysis and WBSF.”

Q 13: LINES 364-366 – The authors say “Interestingly, higher pH values in the SC were not correlated neither with WBSF values or with sensory tenderness values and showed that SC were rated as more tender than SS.”. (…) the samples tested were from fried meatballs made with minced meat obtained from SC and SS and not actual samples of meat slices from SC and SS. (…).

(…)   

Q13.1.2: Thanks for your comment. However, we believe that using acronyms for this reason might, in some cases, lower the clarity of our ideas. Therefore, we did not accept this suggestion.

Q13.3: No criticism about not using individual steaks. I agree with the authors about the problem of using acronyms. My point was just about the importance of clarifying early in the text (and in table titles) that SC and SS refer, respectively to minced samples of shoulder clod and silverside, and not to actual shoulder clod and silverside muscles. Then it would be just a matter of using the acronyms. For other parameters, using muscle samples or minced muscle samples is not critical, but for WBSF and sensory tenderness can be.

A3: Thank you for this input. Even if we consider that this information is clearly explained in different parts of the manuscript, we accepted your comment and we did as suggested.

Q 21: LINES 430-431 – The authors say “higher pH48 values were obtained from the shoulder clod, which resulted in lower L*, a* and b* readings.”. Why do the authors conclude that lower L*, a* and b* readings were consequence of higher pH48 values?

(…)

Q21.1: We appreciate your comment. However, we do not share your position. We believe that this is one of the important conclusions that we got from the MANOVA model, and this is why we did not erase this information from the final version.

Q21.2: My point is just about the strength of your conclusion and not so much mentioning it in “Conclusions”. If, instead of “which resulted” was “which tended to cause” would be less controversial.

A4: Thank you for this comment. We changed this point in previous corrections. You are right when you say that we should only state results that might or might not be related to the factors studied in this manuscript. Therefore, we organized the information in a way that our results express that the factors only tended to affect meat quality traits.

 

Author Response File: Author Response.docx