Allantoin and Tissue Specific Redox Regulation in Mud Crab Scylla serrata under Varied Natural Water Physico-Chemical Parameters
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsReview for the paper “Water physico-chemical parameters induced tissue specific allantoin level and their roles on redox regulation in mud crab Scylla serrata” by Samar Gourav Pati, Biswaranjan Paital, Dipak Kumar Sahoo submitted to “Water”.
The authors conducted a field study to determine the levels of allantoin in the muscle and hepatoancreas of the mud crab Scylla serrata. They constructed a correlation matrix to identify relationships between environmental factors and this parameter. Additionally, they investigated the associations between the allantoin level and various physiological parameters. The study found significant positive correlations between the level of allantoin and stress markers, suggesting that this compound may play a role in the adaptation process to stress conditions. However, the paper requires significant revision.
Recommendations:
L 133. Information about the sex and maturity status of mud crabs should be provided by the authors.
L 169-170. Please rewrite this sentence to improve its clarity.
L 212. The authors mentioned MANOVA, but this statistical method was not described in the Materials and Methods.
L 244. The authors must define the abbreviations for locations.
Sections 3.1–3.5. These sections reiterate the findings presented in Figure 1 and the tables. They should be significantly condensed as they do not offer any new information.
Section 3.6. The authors examined the contribution of different parameters on mud crab physiology under seasonal changes through DFA. However, in this section, they only presented a separation of different tissues without any information about the association between environmental variables and DFA axes. The authors should revise the text or provide additional data on this association by moving the information from the discussion section (Table 6). Supplementing the DFA plot with the results of a multivariate test comparing different tissues and seasons would be beneficial.
L 316-318. This sentence should be rephrased by the authors as it is difficult to understand.
L 344-346. “Thus, the present study provides detailed insights into…” This is a very strong statement considering the real authors’ data, mostly descriptive in nature.
L 446-447. Delete this sentence.
Spatial variations in the allantoin level were detected by the authors, but they did not discuss these results.
The citations and references should be formatted according to the Rules for Authors.
The text contains several grammatical errors and requires revision.
Specific remarks.
L 14. Consider replacing “role on antioxidant homeostasis was investigated” with “role in antioxidant homeostasis were investigated”
L 21. Consider replacing “had negative correlation” with “had a negative correlation”
L 25. Consider replacing “were clearly noticed” with “was clearly noticed”
L 27. Consider replacing “level of allantoin, and activity CAT, GR and was the major contributing factor” with “the level of allantoin as well as activity CAT and GR were the major contributing factors”
L 50. Consider replacing “fosters” with “fostering”
L 73. Consider replacing “not much information available on allantoin induced changes” with “not much information is available on allantoin-induced changes”
L 79. Consider replacing “as an increase in 96% superoxide” with “as a 96% increase in superoxide”
L 92. Consider replacing “of intertidal zone” with “of the intertidal zone”
L 98. Consider replacing “role of allantoin on this crab” with “the role of allantoin in this crab”
L 100. Consider replacing “under the changing environments” with “under changing environments”
L 109. Consider replacing “as sampling site” with “as a sampling site”
L 117. Consider replacing “Each season in India are verifi ed with district change” with “Each season in India was verified with district changes”
L 136. Consider replacing “was transferred to” with “were transferred to a”
L 137. Consider replacing “A 10% (w/v) hepatopancreas and muscle tissue homogenate was” with “10% (w/v) hepatopancreas and muscle tissue homogenates were”
L 144. Consider replacing “in alkaline condition” with “in alkaline conditions”
L 149. Consider replacing “as marker” with “as markers”
L 151. Consider replacing “described in details” with “described in detail”
L 165. Consider replacing “Results” with “The results”
L 167. Consider replacing “two way-ANOVA” with “two-way ANOVA”
L 168. Consider replacing “The difference between the mean values was considered as significant” with “Differences between mean values were considered significant”
L 183. Consider replacing “During summer season” with “During the summer season”
L 184. Consider replacing “sites exhibited 82 and 28% higher significantly” with “sites was 82 and 28% significantly higher”
L 185. Consider replacing “The water pH also recorded 6.38 and 6.1% signifi cantly” with “The water pH was also recorded to be 6.38 and 6.1% significantly”
L 187. Consider replacing “than winter and rainy season” with “than in the winter and rainy seasons”
L 193. Consider replacing “three consecutive day was” with “three consecutive days were”
L 201. Consider replacing “during summer season” with “during the summer season”
L 216. Consider replacing “Collectively, different sampling sites and seasons indicate that muscle tissues were shown 95% decreased TBARS level than hepatopancreas tissue i.e., hepatopancreas > muscle.” with “The TBARS level in muscle tissues was 95% lower than that in hepatopancreas tissues, with the latter showing higher levels.”
L 219. Consider replacing “than winter” with “than the winter”
L 224. Consider replacing “During summer season” with “During the summer season”
L 227. Consider replacing “summer season” with “the summer season”
L 236. Consider replacing “to winter and rainy seasons” with “to the winter and rainy seasons”
L 253. Consider replacing “have also been found” with “has also been found”
L 256. Consider replacing “of hepatopancreas” with “for hepatopancreas”
L 257. Consider replacing “strong positive correlation” with “a strong positive correlation”
L 260. Consider replacing “reached to” with “reached”
L 274. Consider replacing “weak positive correlation” with “a weak positive correlation”
L 278. Consider replacing “very weak negative correlation” with “a very weak negative correlation”
L 292. Consider replacing “strong positive” with “a strong positive”
L 298. Consider replacing “more strong positive correlation” with “a more strong positive correlation”
L 302. Consider replacing “strong positive correlation” with “a strong positive correlation”
L 304. Consider replacing “have better” with “have greater”
L 308. Consider replacing “similar positive correlation” with “a similar positive correlation”
L 352. Consider replacing “crucial role” with “crucial roles”
L 357. Consider replacing “in marine crustacean” with “on marine crustaceans”
L 363. Consider replacing “during summer season” with “during the summer season”
L 369. Consider replacing “rises during summer season” with “rise during the summer season”
L 377. Consider replacing “observed content” with “the observed content”
L 379. Consider replacing “have significant impact” with “have a significant impact”
L 386. Consider replacing “define” with “defines”
L 390. Consider replacing “suggests” with “suggest”
L 403. Consider replacing “concentration of” with “concentration of the”
L 408. Consider replacing “are not supporting the same” with “do not support the same”
L 415. Consider replacing “The lack of” with “The lack of a”
L 422. Consider replacing “in summer season” with “in the summer season”
Comments on the Quality of English LanguageEnglish revisions are required.
Author Response
The authors conducted a field study to determine the levels of allantoin in the muscle and hepatoancreas of the mud crab Scylla serrata. They constructed a correlation matrix to identify relationships between environmental factors and this parameter. Additionally, they investigated the associations between the allantoin level and various physiological parameters. The study found significant positive correlations between the level of allantoin and stress markers, suggesting that this compound may play a role in the adaptation process to stress conditions. However, the paper requires significant revision.
Reply: Thanks for the reviewer for the keen observation on our paper. The comments are highly helpful to improve the ms. We have revised the ms considering all the comments.
Recommendations:
L 133. Information about the sex and maturity status of mud crabs should be provided by the authors.
Reply: Yes, we have added “adult male” to that line.
L 169-170. Please rewrite this sentence to improve its clarity.
Reply: The line is simplified as “Effects of the contribution of different variables such as allantoin, LPx, antioxidant enzymes and small antioxidant molecules on the groups were evaluated employing discriminant function analysis (DFA) according to Garson (2008) and Jennrich (1977).”
L 212. The authors mentioned MANOVA, but this statistical method was not described in the Materials and Methods.
Reply: MANOVA was not conducted for the test so, We have corrected it to “two-way ANOVA”
L 244. The authors must define the abbreviations for locations.
Reply: Yes, we have added abbreviations for location as “GJ- Gujarat, TN- Tamil Nadu, OD- Odisha”
Sections 3.1–3.5. These sections reiterate the findings presented in Figure 1 and the tables. They should be significantly condensed as they do not offer any new information.
Reply: Yes, we do agree, therefore, we have deleted all the redundant lines but kept the lines that describe the changes in %.
Section 3.6. The authors examined the contribution of different parameters on mud crab physiology under seasonal changes through DFA. However, in this section, they only presented a separation of different tissues without any information about the association between environmental variables and DFA axes. The authors should revise the text or provide additional data on this association by moving the information from the discussion section (Table 6). Supplementing the DFA plot with the results of a multivariate test comparing different tissues and seasons would be beneficial.
Reply: Yes, we do agree and data on different axis on Fig. 2. And the association has been described in correlation part. Here the contribution of the variables to study the effects of seasons on the studied parameters especially includes how allantoin is described. So, as per the suggestion we have added few lines to make this section clear.
L 316-318. This sentence should be rephrased by the authors as it is difficult to understand.
Reply: Yes, we have rephrased the sentence as “The mud crabs' muscle and hepatopancreas tissues were tested for allantoin and antioxidant parameters in the summer, winter, and rainy seasons. Each tissue was split into three groups, and the two tissues were taken as a whole into six groups for DFA analysis”.
L 344-346. “Thus, the present study provides detailed insights into…” This is a very strong statement considering the real authors’ data, mostly descriptive in nature.
Reply: Yes, we agreed and modified the line as “Thus, the present study provides possible insights into the modulation of allantoin and its subsequent effects on the studied oxidative stress physiology parameters under the varied physico-chemical parameters of the habitats”
L 446-447. Delete this sentence.
Reply: As per the recommendation we have deleted the mentioned sentence.
Spatial variations in the allantoin level were detected by the authors, but they did not discuss these results.
Reply: Very few studies are available on this topic, therefore, we have written as
Allantoin, being considered as a potential modulator of oxidative stress physiology in a range of organisms [56], seasonally varies in the crab tissues in the present study. Fig. 1 clearly indicates its augmented values in the summer season as compared to the other two seasons. During the summer period all the studied environmental parameters such as pH, temperature, salinity, organic carbon level, Ca and Mg were high and the level of these parameters along with the titre of allantoin was also low in rainy season as compared to the other two seasons. Following the trends, all the above studied parameters were almost observed to be at moderate level. Such data are in agreement with the earlier observation [30, 24, 25]. So, the studied parameters, especially temperature and salinity were able to raise allantoin level in these ectothermic animals in their natural population during summer season. As noted earlier, allantoin is metabolized to ammonia and CO2 in invertebrates [50]. Therefore, under high salt concentration in summer season as compared to the other two seasons could be associated with the low catabolism rate of allantoin. Further detailed study is suggested on this aspect. The crab S. serrata slowly adapts uricotelic excretory metabolism over ammonotelic metabolism as salinity increases in their environment [59, 34]. The reverse mechanism i.e. adaption of ammonotelic nature at low salinity observed in rainy season is also noted in this animal indicating the involvement of salinity in allantoin production. This could be an adapted mechanism the crab follows to produce less ammonia at high salinity. A controlled laboratory experiment to map the allantoin level in these crabs at constant temperature with altered salinity level is suggested to prove this mechanism. The results are confirmation with the rise in oxidative stress markers such as LPx level in the crab during summer season [30, 24, 25]. Therefore, allantoin can also be considered as one of the markers of oxidative stress in invertebrates, as already observed in vertebrates including humans [54, 55, 57].
The citations and references should be formatted according to the Rules for Authors.
Reply: As per the recommendation we have formatted all the citations and references.
The text contains several grammatical errors and requires revision.
Reply: Yes, we do agree, and the corrections are made as per the suggestion by the colleagues of Dr. Dipak K. Sahoo (one the authors) at United State, who speaks English as native language.
Specific remarks.
L 14. Consider replacing “role on antioxidant homeostasis was investigated” with “role in antioxidant homeostasis were investigated”
Reply: Yes, we have replaced the part of sentence with “role in antioxidant homeostasis were investigated”
L 21. Consider replacing “had negative correlation” with “had a negative correlation”
Reply: Yes, we have replaced the part of sentence with “had a negative correlation”
L 25. Consider replacing “were clearly noticed” with “was clearly noticed”
Reply: Yes, we have replaced the part of sentence with “was clearly noticed”
L 27. Consider replacing “level of allantoin, and activity CAT, GR and was the major contributing factor” with “the level of allantoin as well as activity CAT and GR were the major contributing factors”
Reply: Yes, we have replaced the part of sentence with “the level of allantoin as well as activity CAT and GR were the major contributing factors”
L 50. Consider replacing “fosters” with “fostering”
Reply: Yes, we have replaced the word with “fostering”
L 73. Consider replacing “not much information available on allantoin induced changes” with “not much information is available on allantoin-induced changes”
Reply: Yes, we have replaced the part of sentence with “not much information is available on allantoin-induced changes”
L 79. Consider replacing “as an increase in 96% superoxide” with “as a 96% increase in superoxide”
Reply: Yes, we have replaced the part of sentence with “as a 96% increase in superoxide”
L 92. Consider replacing “of intertidal zone” with “of the intertidal zone”
Reply: Yes, we have replaced the part of sentence with “of the intertidal zone”
L 98. Consider replacing “role of allantoin on this crab” with “the role of allantoin in this crab”
Reply: Yes, we have replaced the part of sentence with “the role of allantoin in this crab”
L 100. Consider replacing “under the changing environments” with “under changing environments”
Reply: Yes, we have replaced the part of sentence with “under changing environments”
L 109. Consider replacing “as sampling site” with “as a sampling site”
Reply: Yes, we have replaced the part of sentence with “as a sampling site”
L 117. Consider replacing “Each season in India are verifi ed with district change” with “Each season in India was verified with district changes”
Reply: Yes, we have replaced the part of sentence with “Each season in India was verified with district changes”
L 136. Consider replacing “was transferred to” with “were transferred to a”
Reply: Yes, we have replaced the part of sentence with “were transferred to a”
L 137. Consider replacing “A 10% (w/v) hepatopancreas and muscle tissue homogenate was” with “10% (w/v) hepatopancreas and muscle tissue homogenates were”
Reply: Yes, we have replaced the sentence with “10% (w/v) hepatopancreas and muscle tissue homogenates were”
L 144. Consider replacing “in alkaline condition” with “in alkaline conditions”
Reply: Yes, we have replaced the part of sentence with “in alkaline conditions”
L 149. Consider replacing “as marker” with “as markers”
Reply: Yes, we have replaced the word with “as markers”
L 151. Consider replacing “described in details” with “described in detail”
Reply: Yes, we have replaced the part of sentence with “described in detail”
L 165. Consider replacing “Results” with “The results”
Reply: Yes, we have replaced the part of sentence with “The results”
L 167. Consider replacing “two way-ANOVA” with “two-way ANOVA”
Reply: Yes, we have replaced the word with “two-way ANOVA”
L 168. Consider replacing “The difference between the mean values was considered as significant” with “Differences between mean values were considered significant”
Reply: Yes, we have replaced the part of sentence with “Differences between mean values were considered significant”
L 183. Consider replacing “During summer season” with “During the summer season”
Reply: Yes, we have replaced the part of sentence with “During the summer season”
L 184. Consider replacing “sites exhibited 82 and 28% higher significantly” with “sites was 82 and 28% significantly higher”
Reply: Yes, we have replaced the part of sentence with “sites was 82 and 28% significantly higher”
L 185. Consider replacing “The water pH also recorded 6.38 and 6.1% signifi cantly” with “The water pH was also recorded to be 6.38 and 6.1% significantly”
Reply: Yes, we have replaced the part of sentence with “The water pH was also recorded to be 6.38 and 6.1% significantly”
L 187. Consider replacing “than winter and rainy season” with “than in the winter and rainy seasons”
Reply: Yes, we have replaced the part of sentence with “than in the winter and rainy seasons”
L 193. Consider replacing “three consecutive day was” with “three consecutive days were”
Reply: Yes, we have replaced the part of sentence with “three consecutive days were
L 201. Consider replacing “during summer season” with “during the summer season”
Reply: Yes, we have replaced the part of sentence with “during the summer season”
L 216. Consider replacing “Collectively, different sampling sites and seasons indicate that muscle tissues were shown 95% decreased TBARS level than hepatopancreas tissue i.e., hepatopancreas > muscle.” with “The TBARS level in muscle tissues was 95% lower than that in hepatopancreas tissues, with the latter showing higher levels.”
Reply: Yes, we have replaced the part of sentence with “The TBARS level in muscle tissues was 95% lower than that in hepatopancreas tissues, with the latter showing higher levels.”
L 219. Consider replacing “than winter” with “than the winter”
Reply: Yes, we have replaced the part of sentence with “than the winter”
L 224. Consider replacing “During summer season” with “During the summer season”
Reply: Yes, we have replaced the part of sentence with “During the summer season”
L 227. Consider replacing “summer season” with “the summer season”
Reply: Yes, we have replaced the part of sentence with “the summer season”
L 236. Consider replacing “to winter and rainy seasons” with “to the winter and rainy seasons”
Reply: Yes, we have replaced the part of sentence with “to the winter and rainy seasons”
L 253. Consider replacing “have also been found” with “has also been found”
Reply: Yes, we have replaced the part of sentence with “has also been found”
L 256. Consider replacing “of hepatopancreas” with “for hepatopancreas”
Reply: Yes, we have replaced the part of sentence with “for hepatopancreas”
L 257. Consider replacing “strong positive correlation” with “a strong positive correlation”
Reply: Yes, we have replaced the part of sentence with “a strong positive correlation”
L 260. Consider replacing “reached to” with “reached”
Reply: Yes, we have replaced the part of sentence with “reached”
L 274. Consider replacing “weak positive correlation” with “a weak positive correlation”
Reply: Yes, we have replaced the part of sentence with “a weak positive correlation”
L 278. Consider replacing “very weak negative correlation” with “a very weak negative correlation”
Reply: Yes, we have replaced the part of sentence with “a very weak negative correlation”
L 292. Consider replacing “strong positive” with “a strong positive”
Reply: Yes, we have replaced the part of sentence with “a strong positive”
L 298. Consider replacing “more strong positive correlation” with “a more strong positive correlation”
Reply: Yes, we have replaced the part of sentence with “a more strong positive correlation”
L 302. Consider replacing “strong positive correlation” with “a strong positive correlation”
Reply: Yes, we have replaced the part of sentence with “a strong positive correlation”
L 304. Consider replacing “have better” with “have greater”
Reply: Yes, we have replaced the part of sentence with “have greater”
L 308. Consider replacing “similar positive correlation” with “a similar positive correlation”
Reply: Yes, we have replaced the part of sentence with “a similar positive correlation”
L 352. Consider replacing “crucial role” with “crucial roles”
Reply: Yes, we have replaced the part of sentence with “crucial roles”
L 357. Consider replacing “in marine crustacean” with “on marine crustaceans”
Reply: Yes, we have replaced the part of sentence with “on marine crustaceans”
L 363. Consider replacing “during summer season” with “during the summer season”
Reply: Yes, we have replaced the part of sentence with “during the summer season”
L 369. Consider replacing “rises during summer season” with “rise during the summer season”
Reply: Yes, we have replaced the part of sentence with “rise during the summer season”
L 377. Consider replacing “observed content” with “the observed content”
Reply: Yes, we have replaced the part of sentence with “the observed content”
L 379. Consider replacing “have significant impact” with “have a significant impact”
Reply: Yes, we have replaced the part of sentence with “have a significant impact”
L 386. Consider replacing “define” with “defines”
Reply: Yes, we have replaced the part of sentence with “defines” “defines”
L 390. Consider replacing “suggests” with “suggest”
Reply: Yes, we have replaced the part of sentence with “suggest”
L 403. Consider replacing “concentration of” with “concentration of the”
Reply: Yes, we have replaced the part of sentence with “concentration of the”
L 408. Consider replacing “are not supporting the same” with “do not support the same”
Reply: Yes, we have replaced the part of sentence with “do not support the same”
L 415. Consider replacing “The lack of” with “The lack of a”
Reply: Yes, we have replaced the part of sentence with “The lack of a”
L 422. Consider replacing “in summer season” with “in the summer season”
Reply: Yes, we have replaced the part of sentence with “in the summer season”
Reviewer 2 Report
Comments and Suggestions for AuthorsReport for the authors
Manuscript Number: water-2490930
Title: Water Physico-Chemical parameters induced tissue specific allantoin level and their roles on redox regulation in mud crab Scylla serrata
The manuscript deals with the presence and modulation of allantoin in mud crabs under various environmental conditions – measurements being recorded in animals from the natural populations.
Although there is merit in the data and the amount of work behind the manuscript is impressive, it cannot be published in the present format. The manuscript is difficult to read in parts, it is very wordy and difficult to follow, therefore the scientific value is somehow diluted. I recommend minor revisions.
Very likely, the manuscript would benefit from an overall editing of English language and style. Below are some examples and suggestions:
Abstract
Line 22. ...upregulation of the acidity of major antioxidant enzymes...not sure what this is about?
Introduction
Line 36. .... delete “basically” and continue the sentence: (Igile et al, 2014), as a degraded product…
Line 64 – to Materials and Methods. I suggest this section should be re-written. As an introduction is very convoluted – there should be a very clear section on environmental impact on allantoin levels, then studies on allantoin impact on other physiological processes. Maybe a clear comparison between examples in humans and other animals; but all this should be very structured and justified.
Line 73. Reformulate the sentence, it doesn’t have a verb
Materials and methods
Line 117. Reformulate the sentence, it doesn’t make sense
Results
Line 198. Is 57% an average? Constant for all samples? Need more stats here.
Table 2. Too complex, please consider moving this to supplementary data
Discussions
Line 354. Please try to summarise this huge paragraph – maybe consider creating a diagram, because there is so much text, the interpretation becomes too complex and difficult to follow.
Line 376. Reformulate the entire sentence, it’s too long and confusing.
Line 401. Consider referring to tables and figures to support your explanations.
Line 420. Again, please try to summarise this huge paragraph – maybe consider creating a diagram, because there is so much text, the interpretation becomes too complex and difficult to follow.
Line 458. Reformulate the entire sentence, it’s too long and confusing.
Other comments
Table 3. The legend states that coefficient values without any sign are presented as positive correlations. I can’t see any sign…why mentioning then?
Comments on the Quality of English Language
comments in the section above
Reviewer 3 Report
Comments and Suggestions for AuthorsIn general, the research was well conducted and reported. However, some details are missing in the methods section and some inconsistencies were found in the results section.
I have several detailed comments as outlined below:
Line 137- « 10% (w/v) … tissue homogenate», which was the solvent and the homogenization technique?
Line 143 – the reference (Young and Conway, 1942) should be at the end of the sentence. Which reference material was used to quantify allantoin? Which type of analytical method was used (HPLC, LC-MS, etc)? Please detail.
Line 198- ‘57% higher’, this value is an average, please specify.
Lines 199-208- this text is reversed. Fig. 1 presents in the first place the hepatopancreas (a) and next the muscle (b) data. The text presents first the muscle, and then the HP. This paragraph needs a careful revision.
Lines 221-223- SOD behaved differently: it decreased during summer, but not increased. This text is not in agreement with table 2, and the following discussion.
Line 317- the word ‘for’ might be missing in this sentence («were examined ‘for’ allantoin and antioxidant parameters»)
Line 458-460 – the canonical function ‘6’ was not presented in Table 5 or Table 6. Why is it discussed here?
Comments on the Quality of English Languagenone
Author Response
In general, the research was well conducted and reported. However, some details are missing in the methods section and some inconsistencies were found in the results section.
Reply: Thanks for the reviewer for the keen observation on our paper. The comments are highly helpful to improve the ms. We have revised the ms considering all the comments.
I have several detailed comments as outlined below:
Line 137- « 10% (w/v) … tissue homogenate», which was the solvent and the homogenization technique?
Reply: Here we have used tris buffer for preparing the 10% homogenate which is described in detail in our previous paper “Paital, B., Chainy, G.B.N., 2010. Antioxidant defenses and oxidative stress parameters in tissues of mud crab (Scylla serrata) with reference to changing salinity. Comp. Biochem. Physiol. C 151:142–151. https://doi.org/10.1016/j.cbpc.2009.09.007”
Line 143 – the reference (Young and Conway, 1942) should be at the end of the sentence. Which reference material was used to quantify allantoin? Which type of analytical method was used (HPLC, LC-MS, etc)? Please detail.
Reply: This was measured at 522 nm using spectrophotometer.
Line 198- ‘57% higher’, this value is an average, please specify.
Reply: Yes, we have changed the sentence with specific values as “Irrespective of season and sampling sites, the allantoin content of tissues of mud crab was recorded that indicates hepatopancreas tissue having 322.68, 301.52 & 315.35 % higher allantoin than muscle tissue during the rainy, winter and summer season respectively”
Lines 199-208- this text is reversed. Fig. 1 presents in the first place the hepatopancreas (a) and next the muscle (b) data. The text presents first the muscle, and then the HP. This paragraph needs a careful revision.
Reply: Yes, we have revised the paragraph according to the figure 1 arrangement.
Lines 221-223- SOD behaved differently: it decreased during summer, but not increased. This text is not in agreement with table 2, and the following discussion.
Reply: Yes, we have corrected the sentence as “. However, the SOD activity recorded during the summer season was found to be 22.36 and 21.64% significantly (P < 0.05) lower in muscle and hepatopancreas tissues respectively, compared to the rainy and winter season.”
Line 317- the word ‘for’ might be missing in this sentence («were examined ‘for’ allantoin and antioxidant parameters»)
Reply: Yes, we have corrected that sentence as “The mud crab’s hepatopancreas and muscle tissue were tested for allantoin and antioxidant parameters in the summer, winter and rainy seasons”
Line 458-460 – the canonical function ‘6’ was not presented in Table 5 or Table 6. Why is it discussed here?
Reply: The tables are autogenerated, when we have conducted the DFA in SPSS and presenting all tables are not necessary. There are five tables via six roots, which are need not be presented.
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsThe authors have revised the paper accrding to my coments.
Comments on the Quality of English LanguageMinor.
