Allometric Growth Patterns and Ontogenetic Development during Early Larval Stages of Schizothorax waltoni Regan and Percocypris retrodorslis in Southwest China
Round 1
Reviewer 1 Report
The continued documentation on ontogenetic development and fish larval staging of endemic freshwater species is very important and of high interest for further understanding on the species endemicity dynamics, species ecology and aquaculture.
The methods used are very descriptive and are standard for this type of research.
There are some major issues which need revision before publication.
(1) English language must be improved in the whole document. Some paragraphs are very confused and need re-syntax.
(2) The taxonomic status of the two studied species must be clear and also have to be presented and discussed in Introduction. In Fishbase and throughout the relevant literature Percocypris pingi retrodorsalis Cui & Chu, 1990 and not retrodorslis as referred is given as a synonym of Percocypris tchangi (Pellegrin & Chevey, 1936). Recent papers discuss and present its phylogeny. For example:
· Kottelat, M., 2001. Freshwater fishes of northern Vietnam. A preliminary check-list of the fishes known or expected to occur in northern Vietnam with comments on systematics and nomenclature. Environment and Social Development Unit, East Asia and Pacific Region. The World Bank. 123 p.
· Cui, G.-H. and X.-L. Chu. 1990. Differentiation and distribution of the cyprinid fish Percocypris pingi (Tchang). Acta Zootaxonomica Sinica. 15(1): 118-123. [In Chinese, English summary.]
· Wang M, Yang JX, Chen XY (2013) Molecular Phylogeny and Biogeography of Percocypris (Cyprinidae, Teleostei). PLOS ONE 8(6): e61827.
· Xiao-Qin Xiong & Xing-Jian Yue (2020) The complete mitochondrial genome of Percocypris retrodorsalis (Teleostei, Cypriniformes) in Nujiang River: characterization and phylogenetic position, Mitochondrial DNA Part B, 5:3, 3357-3359, DOI: 10.1080/23802359.2020.1821815
Till now the species is accepted as Percocypris retrodorsalis. So it is crucial for the paper to present which species is studied!!!Moreover, all the relevant literature on taxonomy has to be added.
(3) The species names must be presented in italics in the whole text.
(4) Abstract is handwritten. It should resume the main results of the study in a more appropriate and solid way.
(5) Citations are very poor. Many times, there is a lack of the relevant references in the text, especially in the Introduction. E.g. “Due to the sharp decline of resources, they have been listed as wildlife under second class protection in China.”
(6) Results are presented in a repetitive manner, regarding the allometric relationships of the studied characters. Authors should give also tables with the parameters of the studied allometric regressions such as b, a, P etc.
(7) There is no clear reference in the Material and Methods section about the number of the studied fishes. A summarized table would be very helpful with the means, standard deviations etc for the studied morphometric variables after statistical corrections.
(8) In Discussion section some information is repeated, while it has been already presented in Introduction, see the first paragraph.
(9) A broader literature should be discussed in the Discussion section.
I believe that reviewing on the above points will improve the manuscript status and will add clear and robust information for the two studied species.
Comments for author File: 
Comments.pdf
Author Response
Response to Reviewer 1 Comments
To Reviewer 1:
Dear Sir,
Thank you very much for your useful comments and suggestions. We use red font to show the part that we revised in the manuscript. We revise our manuscript as follows:
- Specific comment: English language must be improved in the whole document. Some paragraphs are very confused and need re-syntax.
Our reply: As suggested, the manuscript had been submitted for English editing at https://www.mdpi.com/authors/english. We believe that the revised version would be more fluent and idiomatic English.
- Specific comment: The taxonomic status of the two studied species must be clear and also have to be presented and discussed in Introduction. In Fishbase and throughout the relevant literature Percocypris pingi retrodorsalis Cui & Chu, 1990 and not retrodorslis as referred is given as a synonym of Percocypris tchangi (Pellegrin & Chevey, 1936). Recent papers discuss and present its phylogeny. Till now the species is accepted as Percocypris retrodorsalis. So it is crucial for the paper to present which species is studied!!!Moreover, all the relevant literature on taxonomy has to be added.
Our reply: Thank you. “Percocypris pingi retrodorsalis” had been modified to “Percocypris retrodorsalis” in the manuscript. and the relevant literature on taxonomy had been added.
- Specific comment: The species names must be presented in italics in the whole text.
Our reply: As suggested, all the species names had be presented in italics in the whole text.
- Specific comment: Abstract is handwritten. It should resume the main results of the study in a more appropriate and solid way.
Our reply: As suggested, abstract was modified.
- Specific comment: Citations are very poor. Many times, there is a lack of the relevant references in the text, especially in the Introduction. E.g. “Due to the sharp decline of resources, they have been listed as wildlife under second class protection in China.”
Our reply: As suggested, relevant references[11-16] were added in the Introduction.
- Specific comment: Results are presented in a repetitive manner, regarding the allometric relationships of the studied characters. Authors should give also tables with the parameters of the studied allometric regressions such as b, a, P etc.
Our reply: As suggested, tables (Table1 and Table2) was added with the parameters of the studied allometric regressions such as n b, a, P etc.
- Specific comment: There is no clear reference in the Material and Methods section about the number of the studied fishes. A summarized table would be very helpful with the means, standard deviations etc for the studied morphometric variables after statistical corrections.
Our reply: As suggested, the reference in the Material and Methods section about the number of the studied fishes were added. As the studied morphometric variables were measured from 0DPHs to 60 DHPs, that is very difficult to add a table, so as to the references cited in this article.
- Specific comment: In Discussion section some information is repeated, while it has been already presented in Introduction, see the first paragraph.
Our reply: The information has been deleted in the first paragraph of Discussion section.
- Specific comment: A broader literature should be discussed in the Discussion section.
Our reply: As suggested, a broader literature has been discussed in the Discussion section (using the “Track Changes” function).
- Specific comment: “Early stage of ontogeny of most fishes is characterized by great variations with great majority of organs and systems” --> “Early stage ontogeny of most fishes is characterized by extensive variability expressed in the morphology and function of many organs and systems”.
Our reply: Thank you. “Early stage of ontogeny of most fishes is characterized by great variations with great majority of organs and systems” had been replaced by “Early stage ontogeny of most fishes is characterized by extensive variability expressed in the morphology and function of many organs and systems”.
- Specific comment: “controlled” --> “is controlled”.
Our reply: Thank you. “controlled” had been replaced by “is controlled”.
- Specific comment: “transformed newly hatched larvae” --> “transform the newly hatched larvae”
Our reply: Thank you. “transformed newly hatched larvae” had been replaced by “transform the newly hatched larvae”.
- Specific comment: “improves” --> “improved”.
Our reply: Thank you. “improves” had been replaced by “improved”.
- Specific comment: The whole paragraph needs better syntax. There is no need to super-analyze terminations as "allometry". Please be short and comprehensive.
Our reply: As suggested, the sentence has been rewritten.
- Specific comment: “Schizothorax waltoni Regan” --> “Schizothorax waltoni Regan, 1905”
Our reply: “Schizothorax waltoni Regan” had been replaced by “Schizothorax waltoni
- Specific comment: Specific comment: The Yarlung Zangbo River is an omnivorous fish???????!!!!! Please re- pharaze and re-syntax
Our reply: As suggested, “Schizothorax waltoni Regan is mainly distributed in the trunk and tributaries of the Xigaze to Milin section of the Yarlung Zangbo River, which is an omnivorous fish, mainly feeding on benthic invertebrates and algae”. had been replaced by“Schizothorax waltoni Regan, 1905, which belongs to Schizohorax Heckel of Cyprinidae, is mainly distributed in the trunk and tributaries of the Xigaze to the Milin section of the Yarlung Zangbo River. This species is an omnivorous fish, and mainly feeds on benthic invertebrates and algae”.
- Specific comment: please delete etc...or add information
Our reply: As suggested, we had deleted “etc.”.
- Specific comment: Add citations “Due to the sharp decline of resources, they have been listed as wildlife under second class protection in China.”
Our reply: As suggested, citation had been added.
- Specific comment: “with the water temperature was constant” --> “with the water temperature to be constant”
Our reply: Thank you. “with the water temperature was constant” had been replaced by “with the water temperature to be constant”
- Specific comment: “of dissolved oxygen” --> “of water dissolved oxygen”
Our reply: “of dissolved oxygen” had been replaced by “of water dissolved oxygen”
- Specific comment: improved with Artemia nauplii, Artemia is written with italics!
Our reply: “improved Artemia nauplii” had been replaced by “improved with Artemia nauplii”
- Specific comment: please give the acronyms in the text, ae. anal fin length (AFL) .... for better understanding. The abbreviation must given in the text apart from the figure legend.
Our reply: As suggested, acronyms was given. “These indicators included the following: the anal fin length (AFL), the body depth (BD), the body depth at the anus level (BDA), the caudal fin length (CFL), the dorsal fin length (DFL), the head length (HL), the head depth (HD), the eye diameter (ED), the pectoral fin length (PFL), the tail length (TAL), the trunk length (TRL), the total length (TL), the snout length (SNL), and the ventral fin length (VFL) (Figure 1)”
- Specific comment: of the 13 morphometric....add space
Our reply: Thank you. “of the 13morphometric” had been replaced by “of the 13 morphometric”
- Specific comment: Please add the total number of the observed larvae each time!!!!
Our reply: the total number of the observed larvae were added at P5.
- Specific comment: light red chromatophores!
Our reply: “light red” had been replaced by “light red chromatophores”
- Specific comment: Chromatophores
Our reply: “no red” had been replaced by “no red chromatophores”
- Specific comment: Add the total number of the studied specimens
Our reply: the total number of the observed larvae were added at P7.
- Specific comment: This information is also given in the Introduction, and is repeated here. Please delete it.
Our reply: As suggested, the information had been deleted at P17.
- Specific comment: That type of information such as appearance of pigmentation, mouth opening etc must be analysed and discussed in detail for the two species and in comparison to other relevant literature of Cyprinidae.
Our reply: As suggested, the information such as appearance of pigmentation, mouth opening etc had been discussed between the two species and other relevant literature of Cyprinidae at P17.
Best regards and wishes!
Author Response File: Author Response.pdf
Reviewer 2 Report
The manuscript entitled ‘Allometric growth patterns and ontogenetic development during early larval stages of two rare fish species in south-west China’ provides some essential data in the field of fish biology. However, the manuscript writing and the presentation of data against publication of article in its current form. Some comments of this manuscript are listed below.
Comments
1. The title is imprecise. Please correct by providing common names and scientific names of the tested fish.
2. Please remove keywords that are already included in the title. You may add further keywords to replace the ones removed if you wish. In this journal, three to ten pertinent keywords need to be added, so that please add more keywords. Generally, keywords should contain words and phrases that suggest what the topic is about. Also include words and phrases that are closely related to your topic. Also use variant terms or phrases that readers are likely to use. The full forms of shortened words and abbreviations should be included as well.
3. Make sure that common names and complete scientific names of species are given on first mention, and check that the spellings are correct and the presentation is consistent; for examples, why is the authority only given for some species and not others? why are some formal names given in brackets whereas others are not? why are some fish given common names only? and why are some fish given scientific names only? In general, full scientific name of species should be given when first mention and then abbreviations of generic name should be used in the other places.
4. In abstract, correct as ‘three periods of the larval growth’, ‘14-23 DPH and 16-26 DPH’ and ‘to develop until 26 DPH’
5. The sentences ‘Allometry is an ordinary characteristic…. abnormal larval growth’ should be separated as paragraph 3 of Introduction part, following with paragraph 4 containing common names and complete scientific names of the tested species.
6. Section 2.1, unit of DO should be given.
7. Carefully correct ‘um’ by ‘μm’
8. Section 2.2, correct scientific name in italics
9. How many tanks? The origin (manufacturer, city, and country) of each instrument, as well as computer software, should be given in brackets.
10. Section 3.1, ‘three larval development phases’
11. Section 3.1.1, the relationships and the second-order polynomial equation were not described here.
12. Define all abbreviations in captions of Figures 3 and 5.
13. Section 3.2, citation errors of Figures in text were found in this part.
14. In the second paragraph of Discussion, correct as ‘10.71 mm, respectively’
15. In the fourth paragraph of Discussion, correct as 14-23 DPH and 16-26 DPH’
16. Lack of precise conclusion. The authors should provide core results from this study.
17. There are several errors relating to the use of spelling, capital/lowercase letters, italics, units, symbols, decimal points, brackets, and scientific names. Please carefully re-check throughout the manuscript.
18. The reference lists were not well checked and contain many errors. The errors found such as scientific names in the article titles (italics), and lack of consistency in writing journal names (full/abbreviated).
Author Response
Response to Reviewer 2 Comments
To Reviewer 2:
Dear Sir,
Thank you very much for your useful comments and suggestions. We use red font to show the part that we revised in the manuscript. We revise our manuscript as follows:
Comments
- Specific comment: The title is imprecise. Please correct by providing common names and scientific names of the tested fish.
Our reply: As suggested, the title has been had been replaced by “Allometric Growth Patterns and Ontogenetic Development During Early Larval Stages of Schizothorax waltoni Regan and Percocypris retrodorslis in Southwest China”
- Specific comment: Please remove keywords that are already included in the title. You may add further keywords to replace the ones removed if you wish. In this journal, three to ten pertinent keywords need to be added, so that please add more keywords. Generally, keywords should contain words and phrases that suggest what the topic is about. Also include words and phrases that are closely related to your topic. Also use variant terms or phrases that readers are likely to use. The full forms of shortened words and abbreviations should be included as well.
Our reply: Thank you. The keywords had been replaced.
- Specific comment: Make sure that common names and complete scientific names of species are given on first mention, and check that the spellings are correct and the presentation is consistent; for examples, why is the authority only given for some species and not others? why are some formal names given in brackets whereas others are not? why are some fish given common names only? and why are some fish given scientific names only? In general, full scientific name of species should be given when first mention and then abbreviations of generic name should be used in the other places.
Our reply: Thank you. Scientific names of species had been given on first mention, then abbreviations of generic name had been used in the other places in the text.
- Specific comment: In abstract, correct as ‘three periods of the larval growth’, ‘14-23 DPH and 16-26 DPH’ and ‘to develop until 26 DPH’
Our reply: the manuscript had been submitted for English editing at https://www.mdpi.com/authors/english. We believe that the revised version would be more fluent and idiomatic English.
- Specific comment: The sentences ‘Allometry is an ordinary characteristic…. abnormal larval growth’ should be separated as paragraph 3 of Introduction part, following with paragraph 4 containing common names and complete scientific names of the tested species.
Our reply: As suggested, the sentences ‘Allometry is an ordinary characteristic…. abnormal larval growth’ had been separated as paragraph 3 of Introduction part, following with paragraph 4.
- Specific comment: Section 2.1, unit of DO should be given.
Our reply: “6.93 ± 0.25 (concentration of water dissolved oxygen)” had been replaced by “6.93 ± 0.25 mg/L (concentration of water dissolved oxygen)”
- Specific comment: Carefully correct ‘um’ by ‘μm’
Our reply: thank you. we have corrected ‘um’ by ‘μm’.
- Specific comment: Section 2.2, correct scientific name in italics
Our reply: We have corrected scientific name in italics.
- Specific comment: How many tanks? The origin (manufacturer, city, and country) of each instrument, as well as computer software, should be given in brackets.
Our reply: As suggested, “5-20 incubation basins were used (about 10000 larvae per container). With the growth of the fish fry, the density of the larvae was gradually decreased from 50 specimens/L to 10 specimens/L.”. The origin (manufacturer, city, and country) of each instrument, as well as computer software, had been given in brackets at P3.
- Specific comment: Section 3.1, ‘three larval development phases’
Our reply: As suggested of Reviewer 3 Comments, this section had been deleted.
- Specific comment: Section 3.1.1, the relationships and the second-order polynomial equation were not described here.
Our reply: Section 3.1.1, the relationships and the second-order polynomial equation were described: “There was a second-order polynomial increase in the TL of S. waltoni from hatching to 60 DPHs (R2 =0·96) (Figure 2)”.
- Specific comment: Define all abbreviations in captions of Figures 3 and 5.
Our reply: As suggested, all abbreviations had been defined in captions of Figures 3 and 5.
- Specific comment: Section 3.2, citation errors of Figures in text were found in this part.
Our reply: Thank you. Section 3.2, citation errors of Figures in text had been corrected.
- Specific comment: In the second paragraph of Discussion, correct as ‘10.71 mm, respectively’
Our reply: Thank you. “10.71 respectively” had been replaced by “10.71 mm, respectively”
- Specific comment: In the fourth paragraph of Discussion, correct as 14-23 DPH and 16-26 DPH’
Our reply: “14-23 DPH 16-26 DPH” had been replaced by “14-23 DPHs and 16-26 DPH s”.
- Specific comment: Lack of precise conclusion. The authors should provide core results from this study.
Our reply: As suggested, the conclusion part has been supplemented.
- Specific comment: There are several errors relating to the use of spelling, capital/lowercase letters, italics, units, symbols, decimal points, brackets, and scientific names. Please carefully re-check throughout the manuscript.
Our reply: Thank you. We had carefully re-checked the manuscript.
- Specific comment: The reference lists were not well checked and contain many errors. The errors found such as scientific names in the article titles (italics), and lack of consistency in writing journal names (full/abbreviated).
Our reply: The reference lists were checked and corrected.
Best regards and wishes!
Author Response File: Author Response.pdf
Reviewer 3 Report
General comments
The submitted article entitle “Allometric Growth Patterns and Ontogenetic Development During Early Larval Stages of Two Rare Fish Species in Southwest China” describes the development during the early life stages of two fish species under culture conditions. The work done is well documented in Material and Methods and the results are well presented. The results are well discussed in this section, but I think they should put more emphasis on species’ adaptations to the environment.
In the Result section the authors expose in detail how the different morphological characteristics are appearing while larvae develop. I suggest including in the MS a new figure showing pictures of the different larval stages observed during the experiments. I think it would help readers to better understand the morphological changes.
I also suggest that the authors consult the article Mendiola et al 2007 "Thermal effects on growth and time to starvation during the yolk-sac larval period of Atlantic mackerel Scomber scombrus" L. Journal of Fish Biology (2007) 70, 895-910. I believe that the statistical analysis of this article can also be applied here and I consider it would be a very positive contribution to the study.
Specific comments are given in the MS attached.
Comments for author File: Comments.pdf
Author Response
Response to Reviewer 3 Comments
To Reviewer 3:
Dear Sir,
Thank you very much for your useful comments and suggestions. We use red font to show the part that we revised in the manuscript. We revise our manuscript as follows:
- Specific comment: The submitted article entitle “Allometric Growth Patterns and Ontogenetic Development During Early Larval Stages of Two Rare Fish Species in Southwest China” describes the development during the early life stages of two fish species under culture conditions. The work done is well documented in Material and Methods and the results are well presented. The results are well discussed in this section, but I think they should put more emphasis on species’ adaptations to the environment.
Our reply: As suggested, the discussion part has been supplemented.
- Specific comment: In the Result section the authors expose in detail how the different morphological characteristics are appearing while larvae develop. I suggest including in the MS a new figure showing pictures of the different larval stages observed during the experiments. I think it would help readers to better understand the morphological changes.
Our reply: As suggested, two new figure showing pictures of the different larval stages observed during the experiments had been added (Figure3 and Figure6).
- Specific comment: I also suggest that the authors consult the article Mendiola et al 2007 "Thermal effects on growth and time to starvation during the yolk-sac larval period of Atlantic mackerel Scomber scombrus" L. Journal of Fish Biology (2007) 70, 895-910. I believe that the statistical analysis of this article can also be applied here and I consider it would be a very positive contribution to the study.
Our reply: As suggested, we had cited this references, from which I got so much，and I need to learn more statistical analysis of this article. However, the statistical method of allometric growth used in this paper refers to many references [5,17,20,21].
- Specific comment: I propose：to improve the growth and survival rates
Our reply: Thank you. “to improve the growth rate and survival rate” had been replaced by “to improve the growth and survival rates”
- Specific comment: In which sense is conductive? Please examples
Our reply: We change this sentence to “Determining relative growth patterns during early growth stages is conducive to the fisheries administration of fisheries and fish farming through promoting normal development patterns in some conditions and improving feeding methods in the case of examined abnormal larval growth”.
- Specific comment: Why 15 °C
Our reply: The water temperature was near optimal for the larvae growth and survival [11-14].
- Specific comment: Please provide reference on why the authors have selected these doses of food. And also why the type of food is change in while the type larvae growth.
Our reply: As suggested, we had added the references.
- Specific comment: I don’t understand the method used. A better explanation is required.
Our reply: The statistical method of allometric growth used in this paper refers to many references [5,17,20,21].
- Specific comment: I think this section is not needed
Our reply: As suggested, this section had been deleted.
- Specific comment: Are the author using a polinomial function?
Our reply: Growth of larvae followed a second-order polynomial curve (between total length and day post hatch). The allometric development was shown as exponential relationship: Y=aXb(between morphological indicators and total length).
- Specific comment: 3.2.1Define b1 and b2
Our reply: As suggested, we defined b1 and b2 in the text: “b1 is b value before the inflection point”. “b2 is b value after the inflection point”.
- Specific comment: More that need I suggest achive,reach
Our reply: Thank you. “need to improve the swimming ability sooner.” had been replaced by “can help to improve the swimming ability”.
- Specific comment: I don’t know what species the authors referring to
Our reply: We had added the species referring to in this sentence.
- Specific comment: Please give more details on it. Adaptions to environment is a very important issue
Our reply: we had added details in the discussion at P19.
- Specific comment: The Phases is very long. Please try to cut it in to 2 sentences or more.
Our reply: As suggested, the phases ha been cut in to 2 sentences. “Based on the developmental features and environmental adaptability of larval fishes, in the production of artificial breeding, it is necessary to provide the environmental conditions required to promote the growth patten in the early stages, and appropriate initial feeding should give priority to the development of important organs so as to effectively acquire exogenous nutrients that integral in order to survive the early dangerous period and avoid death. During the natural reproduction stage of wild fish, it is greatly eco-logically significant to protect their spawning grounds and create a good environment for hatching and early development.”
Best regards and wishes!
Author Response File: Author Response.pdf
Reviewer 4 Report
The manuscript does not have line number and this make revision so hard to define.
The abstract section should start with defining the main objective of the current trial.
At the end of the abstract the authors should highlighted the conclusion of the current trial.
Introduction section sufficient with me but need more cited fresh citation related such as:
https://doi.org/10.22034/IAR.2021.1920885.1129
https://doi.org/10.22034/IAR.2021.1920885.1129
The material and methods missed to define many essential information's as follow:
Information related with parent fish (weight and age)?
Detailed information related with the injection dose and interval, detailed artificial propagation method?
How could the authors check the fertilized eggs?
How many eggs for each female or female weight?
What about the parent tanks and its water quality, fish brooder feed diet …. etc?
Check all scientific names to be in italic forms?
How could the authors enrich artemia and rotifers within larval tanks?
How many larval stocked for each tank? What about water exchange percentage or recycling?
Separated the statistical analysis under separated subtitle? What about testing normality and homogeneity of collected data? What about testing the correlation and regression between tested measurements? How could we predicting measurement with using another? All these concerns should be addressed and calculated.
Separated conclusion under individual title
Author Response
Response to Reviewer 4 Comments
To Reviewer 4:
Dear Sir,
Thank you very much for your useful comments and suggestions. We use red font to show the part that we revised in the manuscript. We revise our manuscript as follows:
- Specific comment: The manuscript does not have line number and this make revision so hard to define.
Our reply: As suggested, we had added the line number
- Specific comment: The abstract section should start with defining the main objective of the current trial. At the end of the abstract the authors should highlighted the conclusion of the current trial.
Our reply: As suggested, we had modified the abstract section.
- Specific comment: Introduction section sufficient with me but need more cited fresh citation related such as:https://doi.org/10.22034/IAR.2021.1920885.1129. We believe that the revised version would be more fluent and idiomatic English.
Our reply: We had cited this fresh citation [15].
- Specific comment: The material and methods missed to define many essential information's as follow: Information related with parent fish (weight and age)?
Our reply: As suggested, information related with parent fish was supplemented. “The total length of the female S. waltoni was 464-628 mm, with a weight of 749-1874 g, while the total length of the male was 352-524 mm, with a body weight of 386-1125 g. The total length of the female P. retrodorslis was 504-940 mm, with a weight of 1540-9300 g, while the total length of the male fish was 350-409 mm, with a weight of 460-750 g. All the females were over 8 years old and the males were over 4 years old, respectively.”
- Specific comment: Detailed information related with the injection dose and interval, detailed artificial propagation method?
Our reply: As suggested, information related with the injection dose and interval, detailed artificial propagation method was supplemented. “Carp pituitary (PG), a luteinizing hormone-releasing hormone analogue (LRH-A2) and human chorionic gonadotropin (HCG) were used to induce articical breeding. The drugs were injected into the abdominal cavity twice. S. waltoni drugs used for the first injection of female contained HCG (300~500 IU/kg) and LRH-A2 (2~5ug/kg), and drugs for the second injection consisted of HCG (800~1000 IU/kg), LRH-A2 (4~7 µg/kg) and PG (5~6 mg/kg). P. retrodorslis: drugs used for the first injection of female included PG (5mg/kg), LRH-A2 (2-5 μg/kg), and HCG (100-300 IU/kg), while the second injection drugs consisted of PG (10 mg/kg), LRH-A2 (5-10 μg/kg), and HCG (1000-2000 IU/kg). Male fish were in-jected only once, and the dose was half that of the female fish. The first and second needles were separated by 24 h, and the fertilized eggs were obtained by artificial dry fertilization.”
- Specific comment: How could the authors check the fertilized eggs?
Our reply: As suggested, information related was supplemented. “We observed the embryo development through microscopic examination and picked out the white eggs.”
- Specific comment: How many eggs for each female or female weight?
Our reply: We believe that this is the content of reproductive biology, which is not covered in this paper.
- Specific comment: What about the parent tanks and its water quality, fish brooder feed diet …. etc?
Our reply: As suggested, information related was supplemented. “Twenty females and 20 males were kept in ponds (10 m long, 4 m deep and 1.5 m high). Each pond provides filtered karst water (100 L/min) from the intake The culture conditions were as follows: 15±1°C (water temperature), 6.93±0.25 mg/L (concentration of water dissolved oxygen), pH 8.08±0.09. The Limnodrilus and special feed for salmon (0.8 # salmon feed produced by Chengdu Gran Company, 58% protein content, sediment feed) were fed together. The feeding amount is 2%~6% of the fish weight, twice a day.”
- Specific comment: Check all scientific names to be in italic forms?
Our reply: As suggested, we had checked all scientific names.
- Specific comment: How could the authors enrich artemia and rotifers within larval tanks?
Our reply: We changed the description. “The feeding of larvae started at 5 DPHs. At first, the feeding was basically carried out with rotifers (Brachionus plicatilis), improved with Chlorella sp., and the concentration was about 30,000 cells/ml.”
- Specific comment: How many larval stocked for each tank? What about water exchange percentage or recycling?
Our reply: As suggested, information related was supplemented. “The incubation basin was a round glass fiber reinforced plastic basin with a diameter of 1.2 m and a water depth of 0.2 m (volume 226 L), and 5-20 incubation basins were used (about 10000 larvae per container). With the growth of the fish fry, the density of the larvae was gradually decreased from 50 specimens/L to 10 specimens/L. Each seedling basin was provided with filtered karst water (100 L/h) from the top”.
- Specific comment: Separated the statistical analysis under separated subtitle? What about testing normality and homogeneity of collected data? What about testing the correlation and regression between tested measurements? How could we predicting measurement with using another? All these concerns should be addressed and calculated.
Our reply: SPSS was used to perform testing normality and one-way ANOVA on the collected data (P=0.05). The statistical method of allometric growth used in this paper refers to many references [5,17,20,21].
- Specific comment: Separated conclusion under individual title
Our reply: As suggested, separated conclusion under individual title was supplemented.
Best regards and wishes!
Author Response File: Author Response.pdf
Reviewer 5 Report
The manuscript suffers from certain omissions. The experimental part is probably correctly performed, but relevant explanations are incomplete. Part of the methodology is not well described There is a need for additional citations, in order to support the included statements. Certain texts are inappropriate, especially in the discussion. The language and style have to be improved significantly; some texts are not understandable, and others are repeated even in one sentence.
Under these circumstances, it is proposed to pass a serious review, in order to be ready for publishing. The authors are encouraged to expose in a better way the methodology, reconsider some statements, and improve the language significantly, so as to re-submit the manuscript.
Some comments are pointed on the attached file.
Comments for author File: Comments.pdf
Author Response
Response to Reviewer 5 Comments
To Reviewer 5:
Dear Sir,
Thank you very much for your useful comments and suggestions. We use red font to show the part that we revised in the manuscript. We revise our manuscript as follows:
- Specific comment: The manuscript suffers from certain omissions. The experimental part is probably correctly performed, but relevant explanations are incomplete. Part of the methodology is not well described There is a need for additional citations, in order to support the included statements. Certain texts are inappropriate, especially in the discussion. The language and style have to be improved significantly; some texts are not understandable, and others are repeated even in one sentence. Under these circumstances, it is proposed to pass a serious review, in order to be ready for publishing. The authors are encouraged to expose in a better way the methodology, reconsider some statements, and improve the language significantly, so as to re-submit the manuscript.
Our reply: As suggested, the manuscript had been submitted for English editing at https://www.mdpi.com/authors/english. We believe that the revised version would be more fluent and idiomatic English. Relevant explanations in Materials and Methods were supplemented at P2-4.
- Specific comment: Please reformulate the text.
Our reply: As suggested, we had reformulated the text.
- Specific comment: There are many repetitions in the text, some sentences are difficult to understand. Please check it thoroughly in terms of style and language.
Our reply: The manuscript had been submitted for English editing at https://www.mdpi.com/authors/english. We believe that the revised version would be more fluent and idiomatic English.
- Specific comment: There are no citations supporting these statements
Our reply: As suggested, we had added the citations [11-16].
- Specific comment: injected twice?
Our reply: Thank you. “twice needles” had been replaced by “injected twice”.
- Specific comment: This means 1.2X0.2=0.24/1000= 240 X10 to 50=
2400 to 5000 specimens per tank right? How many of them were measured?
Our reply: The text had been replaced by “The incubation basin was a round glass fiber reinforced plastic basin with a diameter of 1.2 m and a water depth of 0.2 m (volume 226 L), and 5-20 incubation basins were used (about 10000 larvae per container).”
This means 3.14×0.6m×0.6m×0.2m=0.226m3=226 L, 2000 to 1000 specimens per tank. The number of larvae measured was supplemented. “A total of 955 S. waltoni larvae and 671 P. retrodorslis larvae were measured.”
- Specific comment: specimens per 1 liter?
Our reply: Thank you. “from 50 tails/L to 10 tails/L” had been replaced by “50 specimens/L to 10 specimens/L”.
- Specific comment: Please clarify what means "small"
Our reply: We think it is difficult to clarify, so we delete this sentence.
- Specific comment: approximately per minute or hour?
Our reply: As suggested, “from time to time” had been replaced by “2-4 times per minute”.
- Specific comment: These facts should be supported by data e.g. did You establish exceptions from the general pattern of development? Is development equal for all specimens? Is this the average development?
Our reply: As suggested, we had clarify this method at P4. “These development characteristics were considered to be achieved when at least 50% of the specimens represented this particular stage (n=50-100).”
- Specific comment: It is inappropriate to compare fish growth and development among species from different climatic zones. Temperature is an important factor affecting growth.
Our reply: According to Reviewer 1 Comments "That type of information such as appearance of pigmentation, mouth opening etc must be analysed and discussed in detail for the two species，and in comparison to other relevant literature of Cyprinidae.", we added discussion and analysis in this aspect. “The study of Scomber scombrus L. showed that temperature had a great impact on the growth and development stage of the larvae. We only compared the early development of these fishes at the suitable temperature, respectively”.
- Specific comment: relevant answer to the above comment.
Our reply: “Feeding conditions or genetic elements may help to effectively interpret these diversities. Larger newly hatched larvae have a larger yolk sac, which can provide energy for the metabolism of larvae, help to protect from enemies, establish external feeding, and ensure a higher survival rate of larvae in harsh environments.”
- Specific comment: larvae of both species?
Our reply: Thank you. “two kinds of larvae” had been replaced by “larvae of both species”.
- Specific comment: probably determined
Our reply: “maybe decided by genes” had been replaced by “was probably determinedmaybe decided by genes”.
- Specific comment: Please show a connection between the statements and data.
Our reply: As suggested, “BD and BDA showed positive allometry because of the growth of associated glands and the digestive tract.” had been replaced by “the BD and BDA showed positive allometry because of the inflation of the airbladder and the development of the digestive tract.”
- Specific comment: citations in the last paragraph
Our reply: As suggested, citations in the last paragraph were supplemented.
Best regards and wishes!
Author Response File: Author Response.pdf
Round 2
Reviewer 3 Report
The manuscript is much improved. I have only a few minor comments (see below).
Introduction: In the last paragraph, the authors should make the aim of the study clearer. what does this study contribute to the management of the species? I believe it is now vague.
Figures 3 and 6 are very nice. I suggest the authors also include the length of larvae and juveniles in the caption.
Conclusion: The conclusion should describe the most relevant results. In my opinion, this section should start on line 526.
Author Response
Response to Reviewer 3 Comments
To Reviewer 3:
Dear Sir,
Thank you again for your useful comments and suggestions. We use red font to show the part that we revised in the manuscript. We revise our manuscript as follows:
- Specific comment: Introduction: In the last paragraph, the authors should make the aim of the study clearer. what does this study contribute to the management of the species? I believe it is now vague.
Our reply: As suggested, we had modified this paragraph. “Determining relative growth patterns during early growth stages is conducive to the management of fishery resources and fish farming through promoting normal development patterns and improving feeding methods in the case of examined abnormal larval growth.” and “The present study have aimed to illustrate the early morphological development and allometric growth patterns of S. waltoni and P. retrodorslis larvae and juvenile specimens from the hatching up to 60 days post hatching (DPHs) under controlled conditions, so as to understand the adaptations of the priorities during early development and improve artificial rearing technology.”
- Specific comment: Figures 3 and 6 are very nice. I suggest the authors also include the length of larvae and juveniles in the caption.
Our reply: Thank you. the length of larvae and juveniles in the caption had been supplement (Figures 3 and 6).
- Specific comment: Conclusion: The conclusion should describe the most relevant results. In my opinion, this section should start on line 526.
Our reply: As suggested, conclusion section had been modified accordingly at P18.
Best regards and wishes!
Author Response File: Author Response.pdf
Reviewer 4 Report
No additional comments
Author Response
Response to Reviewer 4 Comments
To Reviewer 4:
Dear Sir,
Thank you again for your useful comments and suggestions.
1.Specific comment: No additional comments.
Our reply: Thank you again.
Best regards and wishes!
Author Response File: Author Response.pdf
Reviewer 5 Report
Dear authors,
the manuscript has been significantly improved concerning language. In this form, the reader can follow the whole procedure, from the experiment to the conclusions. Under these circumstances, it can be accepted.
Author Response
Response to Reviewer 5 Comments
To Reviewer 5:
Dear Sir,
Thank you again for your useful comments and suggestions.
1.Specific comment: the manuscript has been significantly improved concerning language. In this form, the reader can follow the whole procedure, from the experiment to the conclusions. Under these circumstances, it can be accepted.
Our reply: Thank you again.
Best regards and wishes!
Author Response File: Author Response.pdf
