The “True Colours” of Golden Loaches (Teleostei: Cobitidae)

Round 1

Reviewer 1 Report

     Species of Balkan loaches of the genus Sabanajevia have been described on the basis of morphological characters only. Never before have mitochondrial markers been considered, nor assessment of coloration. Using museum materials, Fedorcak et al. considered these characters in association with factors stemming from ecological distribution, especially elevation, to gain new insights into structuring among loach lineages.

     I have only one fundamental question: Why did the authors examine mitochondrial cyt b sequence data as opposed to COI, the more typical marker for DNA barcoding applications?

     To help sharpen the prose, I have made many marks of the manuscript document. I forward that file to the authors. More context-dependent comments follow.

     Introduction. – At line 40, species delimitation is not a tool, but rather a task or objective.  

     Methods. – At line 70, were the specimens fixed in formalin?

     A sentence at lines 72-73 is not needed; it is redundant with the previous sentence.

     At line 104, a supporting citation should be provided for the R statistical environment.

     Discussion. – A passage from lines 325-326 is not a proper sentence.

     At line 334, deeper-bodied fish typify slow-moving waters, not only for loaches. The authors provide an explanation of this – flexibility for maneuvering – that seems implausible. Being deeper-bodied actually makes them less flexible. The deeper form gives them more surface area with which to push against the water for propulsion. Fish living in moving waters need to be more streamlined to reduce drag and maintain position. Being deep-bodied also provides some level of protection from gape-limited predators. These issues might be mentioned.

     At line 338, while enlargement of the gonads may have an effect on body shape, sexual dimorphism is common among fishes - notable examples among stream fishes are provided by the salmonids. What is the spawning behavior of these loaches? Do males fight? Are males more brightly colored than females to attract mates? These issues might be mentioned. The citation provided to support the point that gonadal development explains difference of individual shape is a study of mitochondrial DNA is this really the study that the authors intend to cite?

     At line 369, I am not sure what the “(B” refers to – perhaps this appears by mistake.  

     References. – The citations need cleanup. Species Latin names should be italicized. In the titles of papers cited, all words should not be capitalized. I’ve marked the manuscript to assist with revision.  

     Tables and Figures. – Many of the figures – Figures 1, 3, 5 – are too small to be read properly. Can these be enlarged?

      In the caption for Table 2, is Adonis a software program? If so, a supporting citation should be provided.

     Use of a smaller font for Table 3 would allow it to fit more nicely onto the page.

     The caption for Figure 4 should note that the numbers shown are elevations above sea level in meters.

     In Table 5, can the authors show in bold font which entries are significantly different from zero?

Comments for author File: Comments.pdf

Author Response

Species of Balkan loaches of the genus Sabanajevia have been described on the basis of morphological characters only. Never before have mitochondrial markers been considered, nor assessment of coloration. Using museum materials, Fedorcak et al. considered these characters in association with factors stemming from ecological distribution, especially elevation, to gain new insights into structuring among loach lineages.

     I have only one fundamental question: Why did the authors examine mitochondrial cyt b sequence data as opposed to COI, the more typical marker for DNA barcoding applications?

Most of the molecular data of target genus are found in databases in the form of cytochrome b sequences.

     To help sharpen the prose, I have made many marks of the manuscript document. I forward that file to the authors. More context-dependent comments follow.

     Introduction. – At line 40, species delimitation is not a tool, but rather a task or objective. 

Corrected L40

     Methods. – At line 70, were the specimens fixed in formalin?

explained ((formalin to ethanol procedure) L70

     A sentence at lines 72-73 is not needed; it is redundant with the previous sentence.

deleted

     At line 104, a supporting citation should be provided for the R statistical environment.

Added

R Core Team (2022). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL https://www.R-project.org/

     Discussion. – A passage from lines 325-326 is not a proper sentence.

Corrected 324-327

     At line 334, deeper-bodied fish typify slow-moving waters, not only for loaches. The authors provide an explanation of this – flexibility for maneuvering – that seems implausible. Being deeper-bodied actually makes them less flexible. The deeper form gives them more surface area with which to push against the water for propulsion. Fish living in moving waters need to be more streamlined to reduce drag and maintain position. Being deep-bodied also provides some level of protection from gape-limited predators. These issues might be mentioned.

Explained/Corrected 332-335

By the term manoeuvring, we meant moving between obstacles. In general, taeniform fish are ribbon shaped and laterally compressed. This body shape is good for hiding in small shelters between the macrophytes roots, river particles and wood, etc.

     At line 338, while enlargement of the gonads may have an effect on body shape, sexual dimorphism is common among fishes - notable examples among stream fishes are provided by the salmonids. What is the spawning behavior of these loaches? (L338-341)Do males fight? Are males more brightly colored than females to attract mates? These issues might be mentioned. The citation provided to support the point that gonadal development explains difference of individual shape is a study of mitochondrial DNA is this really the study that the authors intend to cite (deleted L340)?

 

     At line 369, I am not sure what the “(B” refers to – perhaps this appears by mistake.  L371 deleted

 

     References. – The citations need cleanup. Species Latin names should be italicized. In the titles of papers cited, all words should not be capitalized. I’ve marked the manuscript to assist with revision. 

Corrected

     Tables and Figures. – Many of the figures – Figures 1, 3, 5 – are too small to be read properly. Can these be enlarged?

If necessary, they can be sent as a separate pdf file.

In the caption for Table 2, is Adonis a software program? If so, a supporting citation should be provided.

Added (L185)

     Use of a smaller font for Table 3 would allow it to fit more nicely onto the page.

Corrected

     The caption for Figure 4 should note that the numbers shown are elevations above sea level in meters.

Added L218

     In Table 5, can the authors show in bold font which entries are significantly different from zero?

Corrected

Reviewer 2 Report

The manuscript is poorly written. It contains many nouns that are used as adjectives (e.g., fish species). Your shape analyses does not take into account differences in size. Should use a sheared principal component analysis.

See Zootaxa 3647 (1): 101–136

www.mapress.com/zootaxa/ Article

101

http://dx.doi.org/10.11646/zootaxa.3647.1.6

http://zoobank.org/urn:lsid:zoobank.org:pub:D57B95E8-3991-4B30-B22D-83978924D05E

Descriptions of five new species of Metriaclima (Teleostei: Cichlidae)

from Lake Malaŵi, Africa

Comments for author File: Comments.pdf

Author Response

The manuscript is poorly written. It contains many nouns that are used as adjectives (e.g., fish species). Your shape analyses does not take into account differences in size. Should use a sheared principal component analysis.

See Zootaxa 3647 (1): 101–136

www.mapress.com/zootaxa/ Article101 http://dx.doi.org/10.11646/zootaxa.3647.1.6

 

http://zoobank.org/urn:lsid:zoobank.org:pub:D57B95E8-3991-4B30-B22D-83978924D05E Descriptions of five new species of Metriaclima (Teleostei: Cichlidae) from Lake Malaŵi, Africa

 

 

Nonshape variation (due to small differences in size and orientation between specimens) was held by generalized Procrustes analysis (GPA) – function gpagen in Geomorph package. (L103)

Moreover:

L69: The data set (133 specimens) was sex and length (SL) balanced (the length of both sexes did not differ statistically; χ2 = 0.03, d.f. = 1, p > 0.85).

Round 2

Reviewer 2 Report

The authors still did not address my concerns about size differences. They state in line 70 that sizes of the sexes did not differ significantly, but they don't address sizes among species. Some grammar still needs to be corrected. If the size differences are adequately addressed, then the paper should be accepted.

on line 75 change originate to originated

line 48 lineages have (not has) been described

Author Response

The authors still did not address my concerns about size differences. They state in line 70 that sizes of the sexes did not differ significantly, but they don't address sizes among species. If the size differences are adequately addressed, then the paper should be accepted.

Answer:

Lenght differences were checked by Chi quadrat test (L72). There is no statistically significant difference between the mean lengths of the target species (S. balcanica vs. S. bulgarica).

Some grammar still needs to be corrected.

on line 75 change originate to originated - corrected

line 48 lineages have (not has) been described – corrected

The modifications are shown in red.

Author Response File: Author Response.docx