Psammophaga secriensia sp. nov., a New Monothalamid Foraminifera (Protista, Rhizaria) from the Romanian Black Sea Shelf^†

Round 1

Reviewer 1 Report

This is a fairly straightforward description of a new species from the Black Sea belonging to a common shallow-water foraminiferal genus. It s basically OK but includes many minor mistakes and will need quite a major revision to bring it up to a publishable standard.

 

General points

 

1) There is confusion over the use of the word ‘isolate’. From what I understand, this term refers to the DNA extracted from the specimen (i.e., the PCR product). It is not a synonym for ‘specimen’. However, you use it in the same sense as ‘specimen’ in several places (lines 201-202, 222, Table 3, captions for Figs 3 and 5). If you are referring to specimens then I suggest you call them ‘specimens’.

 

2) There is confusion over the upper size limit of your new species and whether or not you are including your earlier Psammophaga sp. (Pavel et al., 2021) in it. The diagnosis states that the new species is less than 800 um long, but the remarks (line 283) give an upper length limit of 1140 um. Presumably, this higher value is because you are including your earlier specimens in the species. However, according to the measurements given in Table 3, the largest specimen of Psammophaga sp. is 1440 um long - not 1140 um, a value that doesn’t appear anywhere in Table 3! So you need to A) decide whether or not you are including the earlier Psammophaga sp. and B) if you are including it, then you should correct the upper size limit. But if you exclude these earlier specimens, then this needs to be made clear.

 

3) Some of the length and width measurements in the test and Table 3 are over-accurate. You cannot measure these dimensions to a fraction of a micron. Please round them up or down to a full micron.

 

4) You include a sequence from a specimen collected in Balaclava Bay, Crimea in your new species. However, you provide very little further information about it, and that only near the end of the manuscript (lines 269-271 and 292-294). Please explain in the methods section whether you collected this specimen yourselves or whether it was collected as part of another project. Also, this sampling locality should be included in Fig. 1.

 

5) There are multiple problems with Figures 2-5. They are poorly presented. I suggest you do the following in order to present the photos in a neat, tidy way,

 - Orientate all of the images so that the long axes of the specimens are horizontal and then crop them so that there is not so much empty space around the specimens.

 - Place the lettering (A, B etc) on the photographs (e.g., in the bottom left corner), not below them (you have no lettering at all in the case of Fig. 3).

 - You can eliminate either Fig. 2A or 2B. They show the same view of the same specimen.

 - You can also eliminate Fig. 2C. It doesn’t really show anything except the broken end of the test.

 - If you follow the suggestions above, then Figs 2B, 3A,B, 4A-C can be combined into one figure. Arrange the photographs so that they fit together neatly in a rectangle.

 - Do the same with Fig. 5. Crop the photos to remove the empty space around the specimens. Arrange the cropped photos neatly in a rectangle, and put the lettering (A-D) on to the photographs

 Minor comments and edits

Line 23-24. The genus Psammophaga, including P. simplora and several undetermined morphotypes, has been reported from different areas of the Black Sea.

25. Are you sure that Psammophaga zirconia is reported from the Black Sea? I thought t was only found in the Adriatic.

53. Delete full stop (.)_ after ‘19’.

60. I suggest changing the word order – ‘was reported to be abundant in Zernov’s  Phyllophora field’

61-62. Please give a reference for the occurrence of P. zirconia in Kazachya Bay. I think this species is only known from the Adriatic.

68, 71. ‘Pavel et al. (2021) – delete comma, add full stop (.) and put brackets around year.

69, 84, 85,95

Minor edits and comments

 

. ‘79 m’ space between number and unit.

107. It is better express this as cm² and call it the cross-sectional area, i.e. - ‘an opening with a cross-sectional area of 78.5 cm².

109. ‘500 um’ (space)

110. I would use the word ‘residues rather than ‘remnants’. If you added to >500 um fraction to the 90-500 um fraction, then the second sentence should read – ‘The >90 um fractions were stored…’ (since this now includes all forams >90 um).

115. Again, space between figure and unit.

117-120. I suggest you rewrite this sentence as follows – ‘Water column parameters including depth……..launched prior to the multicorer in order to avoid any resuspension of the sediment.’

123. There are no keys in Loeblich and Tappan, only generic diagnoses.

p. 4. The table of the top of the page has no caption. Or is this supposed to be part of Table 1 on the previous page? If so, then it makes no sense because the column headings are not the same. Note also that for Station 100C you say the sediment type is ‘Sedimentary rocks’. Do you mean that it’s a hard substrate? If so, this is not consistent with the statement in section 2.3 that you sampled the top 5 cm of sediment.

145 Specimens and isolates are not the same. The term ‘isolate’ refers to the PCR product that is derived from the specimen.

186. ‘Pavel, Kreuter & Holzmann sp. nov.‘ should not be in brackets.

190-191. ‘monothalamous, WITH and elongate……..AND a length of less then 800 um. It HAS a simple…….’

191. Here you say ‘length less than 800 um’ whereas in line 283 you give a maximum length of 1140 um. Where does this figure of 1140 um come from?

191. In the diagnosis and the description I suggest that you give the length/width ratio of the test.

192 (also 209). What do you mean by the aperture being ‘usually titled towards its distal end.’ This is difficult to understand.

192. ‘test wall’ not ‘cell wall’ (the cell is the cytoplasmic body!)

204-205. I’m not sure how you derive the species name ‘secriensia’ from the personal name ‘Secrieru’. The endings of the two words are different. It might be more correct to use ‘secrierui’, although this would be difficult to pronounce.

213. ‘test wall’ (not ‘cell wall’)

214. Delete ‘in P. secriensia’

215-215. Round the measurements to a full micron.

222. Delete ‘isolates’)

222-223. ‘range from 569 to 750 um in length and 230 to 315 um in width’. Please note that if you say ‘range from’ you have to separate the two values with ‘to’ (not a dash).

224-228. This needs to go in the ‘Remarks’ section. Please note also that the dimensions you give here do not correspond to those in Table 3. Here, the length range is for Psammophaga sp. is 719-1440 um and the width 462-775 um.

292. ‘Pawlowski’

Line 23-24. 'The genus Psammophaga, including P. simplora and several undetermined morphotypes, has been reported from different areas of the Black Sea.'

53. Delete full stop (.)_ after ‘19’.

60. I suggest changing the word order – ‘was reported to be abundant in Zernov’s  Phyllophora field’

68, 71. ‘Pavel et al. (2021) – delete comma, add full stop (.) and put brackets around year.

69, 84, 85,95. Please leave a space between the number and the unit.

107. It is better express this as cm² and call it the cross-sectional area, i.e. - ‘an opening with a cross-sectional area of 78.5 cm².

109. ‘500 um’ (space)

110. I would use the word ‘residues' rather than ‘remnants’. If you added to >500 um fraction to the 90-500 um fraction, then the second sentence should read – ‘The >90 um fractions were stored…’ (since this now includes all forams >90 um).

115. Again, space between figure and unit.

117-120. I suggest you rewrite this sentence as follows – ‘Water column parameters including depth……..launched prior to the multicorer in order to avoid any resuspension of the sediment.’

123. There are no keys in Loeblich and Tappan, only generic diagnoses.

p. 4. Please add a caption to the table of the top of the page. Or is this supposed to be part of Table 1 on the previous page? If so, then it makes no sense because the column headings are not the same. Note also that for Station 100C you say the sediment type is ‘Sedimentary rocks’. Do you mean that it’s a hard substrate? If so, this is not consistent with the statement in section 2.3 that you sampled the top 5 cm of sediment.

145 Specimens and isolates are not the same. The term ‘isolate’ refers to the PCR product that is derived from the specimen.

186. ‘Pavel, Kreuter & Holzmann sp. nov.‘ should not be in brackets.

190-191. ‘monothalamous, WITH and elongate……..AND a length of less then 800 um. It HAS a simple…….’

191. Here you say ‘length less than 800 um’ whereas in line 283 you give a maximum length of 1140 um. Where does this figure of 1140 um come from?

191. In the diagnosis and the description I suggest that you give the length/width ratio of the test.

192 (also 209). What do you mean by the aperture being ‘usually titled towards its distal end.’ This is difficult to understand.

192. ‘test wall’ not ‘cell wall’ (the cell is the cytoplasmic body!)

204-205. I’m not sure how you derive the species name ‘secriensia’ from the personal name ‘Secrieru’. The endings of the two words are different. It might be more correct to use ‘secrierui’, although this would be difficult to pronounce.

213. ‘test wall’ (not ‘cell wall’)

214. Delete ‘in P. secriensia’

215-215. Round the measurements to a full micron.

222. Delete ‘isolates’

222-223. ‘range from 569 to 750 um in length and 230 to 315 um in width’. Please note that if you say ‘range from’ you have to separate the two values with ‘to’ (not a dash).

224-228. This needs to go in the ‘Remarks’ section. Please note also that the dimensions you give here do not correspond to those in Table 3. Here, the length range is for Psammophaga sp. is 719-1440 um and the width 462-775 um.

292. ‘Pawlowski’

 

 

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Reviewer 2 Report

The topic could be of interest. However, several points need to be further addressed and developed.

The main concern is about the equivocal attribution and position (both conceptual and formal) you have about the conspecificity of your new studied specimens identified as P. sencrisia and the specimens of Psammophaga presviously studied and left in open nomenclature.

You assess that the two series of specimens have the same shape and exhibit comparable dimensions and consequently you assume that they are identical. However, the measurements you provide (Table 3) do not support your statement because measurements are sensibly different and even without any overlap. You also performed genetic analyses but do not provide any comment about what indications these provide to support or contradict the conspecificity of the two groups of specimens. Finally, you do not formally include the specimens of Pavel et al. 2021 in a synonymic list but maintain for them the name Psammophaga sp. The possible conspecificity is not taken into account when discussing the species distribution that does not include the localities from where samples in Pavel et al. (2021) come from. In conclusion, it is not possible for the reader to understand if you consider all specimens belonging to the same species or not. Data you provide are disputable and the presentation is confusing. The genetic analysis is presented but not fully employed and discussed in order to clarify these issues.

The text is relatively poor and needs to be largely improved and implemented. Consistency must be ensured.

Tables and Figures need to be re-arranged and largely improved to reach a minimal standard for acceptance.

 

Lines 69-72. Poco chiaro e da dove viene il nuovo materiale? È diverso da quello di Pavel 2021???

Lines 78-85: add a few lines to give some eco-bio- information on this basin after its physical properties.

Line 95: do you have really variations along a track measured between the two stations? Or do you have simply the values measured at the two sampling sites? In this latter instance, please, change the text to provide the information correctly. Alternatively, you could arrange a small table summarizing information about all environmental parameters.

Line 97. What do you mean? The adjective sublittoral is not informative. Please, provide information about the (at least qualitative) texture (grain size) and the nature (composition, i.e. siliciclastic, carbonatic, bioclastic…)

Lines 97-98: What do you mean? Sediment? Omit rock and provide a better information…Or sedimentary rock, i.e. a cemented (hard) rock?? In this case, what is the link to the Modioloula habitat? Please explain better.

Lines 109-110: I do not understand this procedure!

       Why did you use a mesh of 500 mm and not larger? if you needed to discard sizes larger than a certain size (not quoted in the text because of the use of a generic “large debris”?) you must use that size! It is also not clear why you separated the material in two sizes to mix it again after separation!!!!

Line 115: solution?   Please, add

Line 117: the part highlighted in blue could be omitted: it is mandatory to measure parametres before sampling!

Line 121: omit.

Line 124-125: was there no tool for measurements associated with the Axiocam 2008???

Line 127. You could include all relevant data in this table: see also comment on line 95. You need to rearrange the table in a different way avoiding a double heading. You could delete some columns (at least Species, but also “Sediment” type, passing this information in the caption and reducing the table to three lines. See text.

Line 172: From the text, it is clear that you only sequenced 6 samples whereas in this table you include your samples whereas it seems that you only accessed data present in GenBank for the majority of theam.

       I suggest you to separate this information in two different tables, or at least to separate clearly specimens you analysed and those for which you only accessed stored data.

Line 175: the change I asked for could be provided in this way, but you need to mark really in bold the specimens!!!

Line 184: I suggest you to omit this taxon here and to include this informal name in the text, as suggested in the Annotated text.

Line 192: what do you mean? I expect an aperture directed toward the distal end. If “tilted”, I expect it is tilted laterally, for instance! Or do you mean directed??? Please clarify.

Line 209: see previous comments

Line 210: is it possible to give insight about the sizes of such grains??

Lines 211-212: did you analyse some of such grains?? Quarts and mica crystals as well as their fragments are very different in aspect, with mica being laminar, that it is not the case for quartz. The colour is also different! I do not understand why you consider these as possible alternatives.

Lines 213-214, also line 194: It is not clear if sediment particle adhere externally to the organic wall or they infill the test being visible from outside, or both. Please, clarify.

Lines 219-220: It is unclear to me how the ingested particles can be placed around the aperture!!!

Line: 226: I think that the authors’ assessment “exhibits comparable dimensions” is not supported by measurements reported in the table 3. Indeed, whereas your newly studied specimens have total ranges of 568-750 mm and 230-371 mm, for length and width respectively, measurements for Psammophaga sp. are larger and not overlapping!!!! I think that if authors want to synonymise the two taxa, they need to support better their choise.

Lines 226-227 and Table 3: measurements provided for the specimens of Psammophaga sp. in the text and in the table are not consistent. Values reported in the text (length 790 µm-1140 µm; width: 460 µm-720 µm) are different from those reported in the table (length: 719-1439 and width 463-775).

Lines 224-228 and Table 3: Usually total ranges, mean values and standard deviations must be provided for each measured parameter.

Line 235: Figure 2: Fig. A and B repeat each other: One of them must be omitted ensuring also a better composition with two images on the same line. Scale bars are not visible

Figure 3: A and B must report the letter A and B, respectively. Measures of the images and scale bars must be standardised within each figure and among figures. Values of each scale bar must be reported in the caption. While the specimen in A is coloured, is the specimen in B treated in a particular way? Or is it untreated?

Figure 4 and caption, Line 250: I suggestd rephrasing of this sentence.

       But I also see the same appearance at the opposite end of the test, at least in A, B left and C left!! Explain.

       In this figure, I suggest to put all the specimen in a line and to indicate each of them with a different letter from A to E, avoiding left and right. I also suggest to standardize the scale bars with other figures. Put letters within images.

Figure 5. Crop all images in order to delete the blackish background and partly enlarge the specimens; put all figures at the same size, and possibly on a single line with specimens isoriented, for instance all with the aperture upward directed. Standardise scale bars. Put letters within images.

Figures 4 and 5. Why are you providing figures of a lot of specimens? You must provide this information in the captions that must be some more than the list of codes for the figured specimens. For instance, you could specify that you show morphological variation of a particular character…

Line 263: So, the distribution only includes localities from where the new samples were collected and not those reported in Pavel et al. as Psammophaga? Do you consider two separate taxa here? But, this is inconsistent with your statements on line 227! If the two sets of specimens are identical, there is only one species and the distribuition is the sum of the distributions of both sets of specimens. However, you need to support strongly the assessments about conspecificity of all your specimens (see comments above).

Lines 267-268: see comments above on this terminology and the real meaning, and re-evaluate/rephrase these statements.

Line 274: you state that the new species is elongate pyriform and for this it differs from P. symphora that is pyriform to ovoidal. However, you figure some specimens that I would define ovoidal as that of Fig. 5C. I suggest to try to quantify these characters (for instance using a ratio in this case) to obtain a stronger and univocal support to your statements. The recognition of an ovoidal morphology is extremely subjective.

Line 276: make this part consistent with information provided in Tab. 2 and in other parts of the text.

Lines 282-283: again! I do not understand why you always use different ranges, thus confusing the reader!!! Be consistent!

Lines 284-286: I suggest you to add a comment such as: For these particular characters, this is probably a different species that needs to be formally described.

Figure 6: what Psammofaga sp. specimens are those listed in the tree? I think they are those studied in Pavel et al. However, you need to be more precise because you have at least three groups of “Psammophaga” specimens: those studied by Pavel et al., those by Sergeeva et al. [21] and those by Gooday et al. [31],

Lines 309-311: I suggest you to provide density evaluations using the same surface unit.

Line 312: a reference is needed supporting this statement.

Line 318: these density values are higher than those you reported above as the highest in the Black Sea where the Crimea peninsula is located!!!!! Please, change these sentences to avoid inconsistency!

Line 319: Is this a peculiarity of such populations???  I think all species in this genus typical thrive as meiobenthos… delete this part and rephrase the whole sentence.

Lines 320-321: I expect hypoxic conditions at these depths in the Black Sea: please, provide some information about this.

Line 323: so, this Psammophaga sp. is a species “different” from P. secriensia you describe! If this is correct, you have to clarify this in the text also in reference to your statement where you define Psammophaga sp. as “identical” to P. secriensia. And remember to state clearly what Psammophaga sp. you consider here (see coment on figure 6).

Lines 323-325: what is the interpretation for this? Any comment??

Lines 334-335: do you have supplementary material?? Not cited in the text! If you have no material, please, delete this part.

References: standardize!

Comments for author File: Comments.pdf

No special comments.

 

Author Response

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Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

The manuscript is much improved but still needs some minor edits and corrections. Please note particularly the point about P. zirconia in the Black Sea.

Line 24. ‘Psammophaga’ in italics                                

64. You cite Gooday et al. (2011) (ref 16) as a source for the statement P. zirconia occurs in the Crimean area, but P. zirconia is not mentioned anywhere in Gooday et al. (2011). I think you got this information from the original description of zirconia in Sabattini et al. (2016). This says that sequences of zirconia are very close to two unpublished sequences of Psammophaga sp. from near Sevastopol. However, this is not the same as the Psammophaga sp. of Gooday et al. (2011), which came from Kazach’ya Bay and branches quite separately from zirconia according to your tree. Please correct the text.

82. ‘461,000’ (close up space)

89. The term abyssal refers to depths greater than 3500 m. A depth of 2212 m would be ‘bathyal’.

95-96. I suggest – ‘thrives in the upper 100 m of the water column, which has lower salinity and reduced density’

97-98. This sentence is a bit confused. I suggest - ‘…from the deeper, denser, anoxic and sulfidic layers that occupy depths below 100-200 m.’  

103. Delete ‘subsequent’.

107-108. Delete ‘measured at both stations’.

115-115. Rephrase – ‘In Table 1, environmental parameters are given for the bay as a whole, rather than for a particular station. The bay is characterized by a substrate consisting of sand and silt.

Table 2. ‘sapela’ (not ‘sapele’)

216. ‘organic TEST wall’ (not ‘cell wall’)

221, 223. Add ‘east’ and ‘north’ to latitude and longitude.

237-238. What do you mean by ‘The organic test wall is fine…’. It doesn’t make sense. Perhaps ‘finely agglutinated’?

239-240. FORAM01_1, which was also been obtained from station 100C, measures….’.

285-286. ‘differs from the type species, Psammophaga simplora,  IN the general shape, WHICH is elongate to ….’

335 and 336. ’86,000’ and ‘116,000’ (, not .)

339-340. ‘Additionally, it can be found in areas with methane seeps…’

Please see above

Author Response

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Author Response File: Author Response.docx

Reviewer 2 Report

The text and the tables and figures have been largely revised and the new version appears as decidedly improved in respect to the first submission.

A couple of points remain problematic: they are reported below with some indications for the authors to solve these issues.

 

New suggested changes (lines refer to the new versions):

Line 98: I suggest rephrasing this sentence as follows: “give rise to persistent anoxic conditions starting from 100-200 m depth” This to indicate that also deeper parts of the basin (i.e. below 200m) are anoxic, that is not clear from your sentence.

Line 234: you changed tilted with bent. However, this change do not solve my issue because both terms imply a certain change of direction and I think that this is not the case with your material where the aperture is simply “directed” distally and being terminal and on the same axis of the length, is not tilted, bent or curved!

Lines 236-237: Inclusions consist of dark and transparent particles, the latter ones likely composed of quartz or mica.

This was a point marked in the previous round and your answer is not appropriate to support your statement. Ask other more qualified people!

My previous comment: ?? Quarts and mica crystals as well as their fragments are very different in aspect, with mica being laminar, that it is not the case for quartz. The colour is also different! I do not understand why you consider these as possible alternatives.

Your answer: We showed the pictures to my fellow mineralogists and they suggested the presence of such minerals.

As I already told you, the two minerals are very different, and it is impossible to misidentify one for the other. Quartz is never laminar alike micas are!

A further possibility is that the English translation is the source of this issue. I mean, you have both minerals. In this latter case, I suggest you to change the sentence as I reported in the commented PDF file.

Lines 351-353: How to explain this close relationship between this Mediterranean species and two species from Antarctic-subAntarctic area? A sentence with a possible interpretation? Or just to consider this point?

References: some journals are reported in fool whereas other ones are abbreviated, some with the inclusion of full stops at the abbreviation point, some non! Please, standardize.

Comments for author File: Comments.pdf

No special comments.

Author Response

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