Reproductive Behavior and Sexual Patterns in Two Cales, Heteroscarus acroptilus and Olisthops cyanomelas (Odacidae) at Rocky Reefs in Temperate Australia

Round 1

Reviewer 1 Report (New Reviewer)

I think this paper provides some new information about the reproductive behavior of two labrids found in the Pacific.  While providing new, heretofore unknown information about species of any kind is valuable the authors do not make the case in the introduction or discussion as to why this information is important. Is it important to coral reef fish managers? Who will benefit from this information being discovered and published?

I have provided a scanned copy of the paper with annotated comments i wrote on the paper at the location where i wanted to make a comment or suggestion. A few of the more important items i want to highlight are mentioned below:

Page 3, Section 2.2, the whole narrative, the way it is written, is confusing. I don't know if there is a way to present all of this narrative information in a clearer manner, perhaps a table?

Line 227 - what are the '23 species of other labrids' mentioned here? are they listed somewhere?

Page 7, Section 4.1,Line 231 - the mention of histological observations of a different species of labrid (Odax pullus) may not tell anything about the observations in this study on the two subject species, and this tactic seems like a fallacious tack to take. I think this section should be reorganized/rewritten. i put a suggestion for a new beginning sentence to replace the current 'On the other hand....'.

Lots of comments about the last two paragraphs of discussion around the subject of evidence for protogyny/gonochorism. Please address as many of these as you can.

Overall i think this paper will provide some info that is of interest to some local area fish biologists. But the paper needs to be re-organized and written better. The whole thing seems to be more of a summary of a collection of independently conducted random dive trip observations strung together into a data stream and analyzed. This is the reason that when asked about the sample/study design i replied Not Applicable, because it doesn't seem like there is one, its just opportunistic combining of random data collection. Perhaps try to rewrite the study methods section.

For the above reasons, i recommend that this paper be revised and resubmitted when these issues are addressed.

Comments for author File: Comments.pdf

Overall the quality of English language usage is acceptable. but there are times where the construction seems a little stilted or off. There are multiple occurrences of this, one example that i identified is in the third paragraph of page 8. The manuscript could be improved with a thorough review of grammatical usage.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report (New Reviewer)

The authors present original and never-before reported information for 2 species of labrid relevant to reproductive pattern and spawning behavior in fishes. This information is helpful to others studying sexual patterns in fishes and will be beneficial as a published dataset. The Introduction would benefit from a bit more background on the importance of these species and also expand upon why the dataset is so limited (i.e., this study was performed peripherally to other ongoing work or whatever the justification was for not presenting a more thorough investigation). Specific comments are attached

Comments for author File: Comments.pdf

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

The behavioural material is low key but adequate and will interest students of reef fish reproductive behaviour although the video material should be trimmed down to two presentations.

The Gonadal tissue and sexual systems of Heteroscarus acroptilus section appears to be based on 4 specimens two of which are males, For Olisthops cyanomelas there are 18 specimens, with 8 males, two of which had IP colouration. This sample size is totally inadequate as a basis for determining paths of sexual ontogeny in labriform fishes. Testis morphology that display evidence of ovarian structure are not evidence of  a past female function (Taylor et al 2020 Coral Reefs 37 (4), 1197-1208, Hamilton et al (2008) Coral Reefs 27, 153-163} Here examination examination of 179 gonads and macroscopic analysis of 2602 gonads of a labrid fish confirmed gonochorism although testis displayed an ovarian lumen. Moreover bisexual gonads are not always indicative of a hermaphroditic sexual ontogeny (Lowe et al (2021) Journal of fish biology 99 (4), 1348-1363).

 The sample sizes in this section preclude publication.

Reviewer 2 Report

The manuscript (MS) focused primarily on the courting and mating of two different labrid species using visual census, and secondarily on the hermaphrodite behavior dissected from the gonad histology. The previous studies showed significantly these sexual changes for the labrid species as understood from the literatures. 

My consideration is that there is one visual census among the tens visual records as emphasized in the Material and Methods.

Looking at one record (photos captured from the videos), a lot of visual history for the courting and mating of the labrid were resulted in a pattern generalized in the MS. To enhance the pattern, I recommend strongly that the authors would overview all records containing such behaviors  to schematize illustration of them or digitize the video data of them to make statistics.

The following histories are that we observed from the same video of each species are too much. 

Line 146-154

Line 169-176

As further videos are available, each behavior (Line 146-154, and Line 169-176) could be tabulated in a Table by checking in or out for each season or sampling month of all the records. 

Such data will enhance the data and result quality for generalizing the patterns. 

The MS needs inbetween-minor and major revision regarding the comments. If not, I recommend that the MS could be a short note by shortening the text and changing the format. 

Reviewer 3 Report

·         No clear objective has been established. On one hand, authors say, “we aimed to reveal the reproductive behavior of two odacids”…, then they say “this is the first report of the aggressive, courtship, and spawning behavior of odacids…then they say “we clarified the sexual pattern of the two odacids…”, then they say “we investigated the fit of the sexual pattern and mating behavior of the odacids”…then, they say “we compared the similarities and differences in the reproductive behavior and sexual pattern. So, I strongly suggest authors to integrate all these statements into a sentence explaining the objective clearly.

·         In the methodology, it is required authors explain which method is going to be addressed to survey a given aspect of reproduction.

·         Map must be improved by adding geographic coordinates and compass Rose.

·         In Methods, when author say “we measured total length and body weight”, how did they do that and using what? Centimeters, millimeters, grams?

·         Which methodology was followed for histology? Quote any procedure!

·         Which scientific literature was used to classify gonad phases?

·         I did not find any subsection labeler as Data analyses. In this, authors need to mention which variables used and which statistical method was used to compare those variables.

·         Regarding the Results, I find the narrative way too general and no quantitative outcomes were recorded from the observations (times fish do this or that)

·         Regarding the gonad histology 22 fish is a very low number as to identify key aspects such as mature stages or even spawning season. It would be necessary to sample a year in order to properly identify spawning season. The only thing possible with 22 fish is a very limited description.

·         In general terms the manuscript as it stands requires substantial changes and clearly define a basic objective, which is not right now. I urge authors to remove procedures which are not conclusive. The observations done underwater have to be properly documented and explained quantitatively instead of just qualitatively.

·         In the Introduction, it is necessary authors to state that nothing is known of the reproductive aspects of these fishes.

·         What authors can do with this limited sampling is just describing basic reproduction, but it is not possible to investigate sexual pattern or even spawning season or maturation just with 22 fish sampled.

Consequently, I suggest rejection

Reviewer 4 Report

In this study, the reproductive behavior and sexual patterns in two species of Cales have been studied. While the study is interesting, the results are presented in a snapshot and may not give comprehensive conclusions as other year-throughout studies in fish reproduction. Also, some main parameters of reproductive physiology are missing from the results (e.g. the gonadosomatic index (GSI), quantitative measures of behavioral traits, cortisol levels etc.). The Sample size is also very small and accurate results may not be achieved. I am afraid the manuscript is not suitable for publication in its current form.

Best regards,