Chronic Effects of Diazinon® Exposures Using Integrated Biomarker Responses in Freshwater Walking Catfish, Clarias batrachus
, Azubuike V. Chukwuka 2, Dip Mukherjee 3, Lipika Patnaik 4, Susri Nayak 4, Kishore Dhara 5, Nimai Chandra Saha 6 and Caterina Faggio 7,*
Round 1
Reviewer 1 Report
The original article "Chronic effects of Diazinon® exposures using integrated biomarker responses in freshwater walking catfish, Clarias batrachus" addresses a subject of great interest: the long-term impact on fish of an extensively used pesticide.
It deserves to be published in Applied sciences, however, there are several points in which improvements should be made to strengthen the manuscript.
In the introduction it would be beneficial to add some information about the risks to human health.
Lines 26, 27, 31, 32: The abstract contains several abbreviations that are defined only later. Please replace the abbreviations by full length words or add the definitions. The Instructions for Authors mention “Abbreviations should be defined the first time they appear in each of three sections: the abstract; the main text; the first figure or table".
Line 38: I suggest adding the growth, biochemical and endocrine parameters to Keywords.
Line 59: Please add the definition of PBPK/PD.
Line 65: Please remove the word “While” in order to improve the clarity of the statement.
Line 79: Please add the definition of IBR.
Line 103: Please add (APHA, 2012) to the References.
Line 109: Please add the definition of DO.
Line 112: Please add the definition of QA/QC.
Line 130: Earlier work is mentioned, but there is no citation.
Lines 156-203: Please use subscript/superscript in order to improve the clarity of the information.
Line 164: Please add the definition of TEC.
Line 176: What was the procedure of liver weight measurment?
Line 229: The concentrations of diazinon (0.53 mg/L and 1.06 mg/L) referred in this figure and in the following ones are different compared to the ones mention in the Abstract and Materials and Methods (0.64 mg/L and 1.28 mg/mL); please clarify.
Line 230, 268, 293, 315: Please add the explanation regarding the number of * depicted in each figure.
Line 237, 335: The definitions of K and SGR were already introduced.
Lines 272, 277, 299, 303, 366, 380, 407: Please use italics for the name of the species.
Line 280: Please add the definition of HPT.
Lines 298-299: TSP and TSG have already been introduced.
Lines 297, 299, 319, 367: The figures are mentioned in the text in an inconsistent style, please harmonize.
Line 308: Please add the definition of AChE.
Line 343: Please add the definition of TP.
Line 373: The IBR values are shown in Figure 7.
Line 405: The title of the vertical axis of Figure 7a is missing. The parts a and b of Figure 7 should be harmonized regarding the chart style.
Line 483: The reference should also contain information regarding the encyclopedia.
Author Response
Reviewer 1
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1 |
In the introduction it would be beneficial to add some information about the risks to human health. |
Information added: Significant risks of cancer, lung lesions and cytogenetic effects in humans under chronic or prolonged exposures to diazinon have also been demonstrated. (Lines 63, 64) |
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2 |
Lines 26, 27, 31, 32: The abstract contains several abbreviations that are defined only later. Please replace the abbreviations by full length words or add the definitions. The Instructions for Authors mention “Abbreviations should be defined the first time they appear in each of three sections: the abstract; the main text; the first figure or table". |
Abbreviations defined in all sections.
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4 |
Line 38: I suggest adding the growth, biochemical and endocrine parameters to Keywords. |
Keywords added. |
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5 |
Line 59: Please add the definition of PBPK/PD. |
definition of PBPK/PD added: Physiologically Based Pharmacokinetic/Pharmacodynamic. Lines 66 |
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6 |
Line 65: Please remove the word “While” in order to improve the clarity of the statement. |
the word “While” is removed. Line 73. |
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7 |
Line 79: Please add the definition of IBR. |
definition of IBR is added. Line 85. |
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8 |
Line 103: Please add (APHA, 2012) to the References. |
APHA, 2012 reference added. Line 110. |
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9 |
Line 109: Please add the definition of DO. |
definition of DO is added in table 1. |
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10 |
Line 112: Please add the definition of QA/QC. |
definition of QA/QC is added: Quality Assurance/ Quality Control. Line 116. |
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11 |
Line 130: Earlier work is mentioned, but there is no citation. |
Citation is added. Line 135. |
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12 |
Lines 156-203: Please use subscript/superscript in order to improve the clarity of the information. |
subscript/superscripts are used (Lines 156-208). |
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13 |
Line 164: Please add the definition of TEC. |
The term “Total White Blood Count (TWBC)” is used in place of TEC. (line 161) |
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14 |
Line 176: What was the procedure of liver weight measurment? |
Liver weights were recorded to the nearest tenth of a milligram. (Line 177). |
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Line 229: The concentrations of diazinon (0.53 mg/L and 1.06 mg/L) referred in this figure and in the following ones are different compared to the ones mention in the Abstract and Materials and Methods (0.64 mg/L and 1.28 mg/mL); please clarify. |
concentrations of diazinon (0.64 and 1.28 mg/L) are corrected. Line 238. |
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16 |
Line 230, 268, 293, 315: Please add the explanation regarding the number of * depicted in each figure. |
explanation regarding the number of * depicted in each figure are added: *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001. Lines 240, 276, 303, 324. |
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17 |
Line 237, 335: The definitions of K and SGR were already introduced. |
definitions of K and SGR were removed. Lines 237, 335. |
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18 |
Lines 272, 277, 299, 303, 366, 380, 407: Please use italics for the name of the species. |
italics used for the name of the species. |
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19 |
Line 280: Please add the definition of HPT. |
definition of HPT is added. Line 286. |
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20 |
Lines 298-299: TSP and TSG have already been introduced. |
Definitions of TSP and TSG are removed. Lines 300. |
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21 |
Lines 297, 299, 319, 367: The figures are mentioned in the text in an inconsistent style, please harmonize. |
The figures are mentioned in the text are harmonized. |
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22 |
Line 308: Please add the definition of AChE. |
definition of AChE added- Acetylcholinesterase. Line 314. |
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23 |
Line 343: Please add the definition of TP. |
definition of TP is added. Line 349. |
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24 |
Line 373: The IBR values are shown in Figure 7. |
Corrected. Line 379. |
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25 |
Line 405: The title of the vertical axis of Figure 7a is missing. The parts a and b of Figure 7 should be harmonized regarding the chart style. |
The title of the vertical axis of Figure 7a is added. The parts a and b of Figure 7 are harmonized. |
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26 |
Line 483: The reference should also contain information regarding the encyclopedia. |
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Reviewer 2 Report
This is a well-written manuscript and the investigation is novel especially with regards to the influence of pesticides on aquatic animals
Author Response
Reviewer 2
Comments and Suggestions for Authors
This is a well-written manuscript and the investigation is novel especially with regards to the influence of pesticides on aquatic animals
Reviewer 3 Report
Saha and colleagues illustrates a study aimed to assess the chronic toxic impact of diazinon on haematological (Hb, HCT, MCH), growth (K-factor, HSI, SGR), biochemical (serum glucose, protein), and endocrine (growth hormone, T3 and T4) parameters in Clarias batrachus. In addition they also evaluate the gross effect of diazinon on fish using Integrated Biomarker Response (IBR) as a summarizing index. Methods are appropriate and properly described. Results are consistent with the conclusions and Discussion is properly focused without useless speculations. In my opinion, the paper can be accepted as it is, except for a check of several typos, references to figures, and few other points here highlighted listed.
Section 2.5. The number of fish used in the study is not clear. Authors stated that “ten acclimatized fingerling fish were transferred to experimental tanks in four groups for haematological, biochemical, growth, and endocrine parameter assays”. This means ten fish for haematological analysis, ten for biochemical one, and so on, or ten for each experimental group (control, T1 and T2…but in this case there are just three groups). This point needs to be better addressed. I also suggest to indicate the number of animals used for each analyses.
line 79. Please give here the full name for IBR.
line 209. The section title “Results” should be "Results and Discussion".
line 373. It should be Fig. 7.
Author Response
Reviewer 3
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1 |
Section 2.5. The number of fish used in the study is not clear. Authors stated that “ten acclimatized fingerling fish were transferred to experimental tanks in four groups for haematological, biochemical, growth, and endocrine parameter assays”. This means ten fish for haematological analysis, ten for biochemical one, and so on, or ten for each experimental group (control, T1 and T2…but in this case there are just three groups). This point needs to be better addressed. I also suggest to indicate the number of animals used for each analyses. |
In a randomized design, acclimatized fingerling fish (n=10) in experimental tanks (350 L capacity) were setup in four replicates. To assay for haematological, biochemical, growth, and endocrine parameters, 3 individuals were sampled from each exposure concentration at day-15, 30 and 45. Total 120 fish are used in this long term experiment.
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2 |
line 79. Please give here the full name for IBR. |
full name for IBR is given. |
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3 |
line 209. The section title “Results” should be "Results and Discussion". |
Corrected as "Results and Discussion". |
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4 |
line 373. It should be Fig. 7. |
Corrected as Fig. 7. |
Round 2
Reviewer 1 Report
Dear Authors,
Congradulations for your comprehensive study.
Please also modify the concentration in figures 2, 3, and 4 from (0.53 mg/L and 1.06 mg/L) to (0.64 and 1.28 mg/L).
Author Response
Please also modify the concentration in figures 2, 3, and 4 from (0.53 mg/L and 1.06 mg/L) to (0.64 and 1.28 mg/L). DONE
