Molecular Characterisation and Phylogenetic Analysis of Dermatophytic Fungi Isolated from Tinea Capitis in Northwest Nigeria Using Sequence of the 28S rRNA

Round 1

Reviewer 1 Report

This interesting manuscript by Dr.  Hussain Yahaya Ungokore and collaborators provides a novel method of effective and sensitive detection of  dermatophytes lodged in human skin scale. Authors identified for the first time the emerence of Trichophyton eriotrephon, Trichophyton bullosum, Trichophyton simii, Ctenomyces 35 serratus and Trichophyton benhamiae dermatophytes from Tinea capitis in Nigeria.

I think that the manus is well illustrated and written in a good language.

Below my suggested changes that could improve the manus:

 

Abstract

Line 15: “molecular microbiology” could be biomedicine

Line 18: “the use of 28S rDNA genes are needed but with certain limitations.” could be  “the use of 28S rDNA genes has certain limitations.”

Line 22: “In this study,  a set of primers…”

Line 35: “ T. eriotrephon, T. bullosum, T. simii, T. benhamiae  and Ctenomyces serratus”

Keywords:

I think you could add “ Trichophyton” and “Ctenomyces” as keywords

 

Introduction

Line 62: “scaly” should not be in bold

Author Response

Dear Reviewer,

Do find attached a copy of my responses

Author Response File: Author Response.pdf

Reviewer 2 Report

The authors describe an investigation of the incidence od several fungal dermatophytes as causing agents of Tinea capitis in a particular region of Nigeria. They use 28s rRNA gene sequencing and database comparisons to confirm the identity of the different isolates and to stablish the level of their intraespecific variability.
The work has been properly planned and performed and the results are clear and well interpreted. Although this piece of work does not represents a significant advance in the field, it is very important in terms of promoting research, knowledge and public health in non-developed african regions.

May be it would be useful to compare these results with the molecular characterization and sequencing of the ITS regions commonly used in fungal molecular taxonomy and phylogeny.

Sequences of primers used for the amplification of 28s rRNA should be detailed.

My main concern is the akwards use of the english lenguage, sometimes it makes dificult the comprehension of the ideas. The text should be in depth corrected by some professional.

Author Response

Dear reviewer,

Do find attached my response

Author Response File: Author Response.pdf

Reviewer 3 Report

The work entitled " Molecular Characterisation and Phylogenetic analysis of dermatophytic fungi isolated from Tinea capitis in Northwest Nigeria using sequence of the 28S rRNA” has an interesting approach for publication in Microbiology reserarch. But, there are some questions of form that should be taken into account prior to consider this article for publication.

I enclose my suggestions for consideration by the authors.

 

• Introduction:

lines 78-79, the authors reflect the advances in molecular biology for the detection and identification of dermatophytosis. It would be interesting for the authors to make a small reflection on the advances in the identification of these infections directly from nail fragments, since it is possibly the most complex structure of the skin for these diagnosis.

I attach some works that can help the authors in this topic.

doi: 10.7547/0003-0538-104.3.233.

doi: 10.1080/14787210.2018.1544891.

 

• Methods:

In this section the authors should make major modifications

The authors must reflect the approval references of the Ethics Committee, and that the participants gave their informed consent.

How many subjects were sampled, and their personal characteristics.

Although sections 2.3 and 2.4 are described in previous works or by the manufacturer's instructions,  the authors should make a brief description of these procedures. They have to make the following issues clear:

• how the sampling was done.
• DNA extraction was extracted directly from the sample or from the culture micelles.
• which Primer were used.

 

• Discussion:

As I have commented previously, there are studies that describe the extraction of DNA directly from human samples (nails) for the identification of the infecting agent. Do the authors consider this possible in Tinea capitis? These items should be addressed.

• References:

some do not meet the publication standards, this must be corrected.

Author Response

Dear Reviewer, 

Find attached a copy of my response

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

The authors of the work entitled "Molecular characterization and phylogenetic analysis of dermatophyte fungi isolated from Tinea capitis in northwestern Nigeria using the 28S rRNA sequence", they did a great job to improve the manuscript but there are little things to clarify. Which take a lot of importance because It is a research that does not provide a significant advance in the study area, it may be very important to improve knowledge and public health, which can make it interesting for publication in Microbiology research.

 

The characteristics of the population should be more detailed ... mainly the recruitment protocol.

The take sampling protocol must be detailed, the bibliography supports that it is a crucial process to obtain truthful results.

 

I would like to propose again that the authors reflect on these two items that I detail in the previous review.

• • Introduction:

lines 78-79, the authors reflect the advances in molecular biology for the detection and identification of dermatophytosis. It would be interesting for the authors to make a small reflection on the advances in the identification of these infections directly from nail fragments, since it is possibly the most complex structure of the skin for these diagnosis.

I attach some works that can help the authors in this topic.

doi: 10.7547/0003-0538-104.3.233.

doi: 10.1080/14787210.2018.1544891.

• • Discussion:

As I have commented previously, there are studies that describe the extraction of DNA directly from human samples (nails) for the identification of the infecting agent. Do the authors consider this possible in Tinea capitis? These items should be addressed.

As previously mentioned, the work does not provide great advances, but reflecting on these issues and how these results can help improve previous researches, as well as reflect the limitations themselves, makes the manuscript more relevant.

Author Response

 

 

 

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

This study investigated the sequence characteristics of dermatophytic fungi causing Tinea capitis from Northwest Nigeria. The variation of sequence within the species and the distance between the species were analyzed and the phylogenetic analysis was performed based on the 28S ribosomal RNA region. The subject is important but several limitations show that this manuscript is not ready to be published in the journal.

The most important limitation is that the experimental design and analysis are not sound to support the author’s opinions.

The authors obtained the sequences from 32 strains from 8 species. The number of the sequence is too small to check molecular characteristics.

The identification method is also unclear, which may be associated with the results that some sequences were more similar to it of the different species than it of the same species.

Also, the phylogenetic analysis did not contain the reference sequences (Fig. 2) or the sequences generated from this study did not cluster with the reference sequence. This is ridiculous!

In the Discussion, the authors discussed many points that did not cover in the Results part. For example, the age or sex of the patients (L198-202) and the host type (zoophilic and anthropophilic species) (L243-246) were discussed without the result from the analysis.

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Reviewer 2 Report

This manuscript by Dr.  Ungokore Hussain Yahaya  et al. aims to identify fungal DNA from clinical samples. In particular, authors identified, several strains belonged to Tinea genus using the 28S rDNA.

There are several issues with the references; e.g. they are not numbered in order of appearance and the reference list is in Times New Roman. I suggest to use Palatino as the text.

 

Below suggested changes:

Abstract

Lines 18-19: This sentence seems incomplete “It’s ranking in identification, phylogenetic analysis and taxonomy of 32 strains of 8 dermatophyte species.” Please rephrase.

Line 22: rDNA

Line 23-25: Please specify genus

Line 29:  emergence -> evidence

 

Keywords:

I think you could add Tinea as a keyword.

 

Introduction

Line 34: It seems in a different format “ The incidence, distribution and control of dermatophytes has progress significantly”

Line 38: “as such not much” unclear what you mean

Line 39: Gene repository. Please, add a hyperlink

Line 51: “scaly” in bold?

Line 58: Trichophyton in italics

Line 47: “hosts.20”. Are the references numbered?

 

Methods

Line 94: delete “(2011)”

Line 96: Why did you re-amplify the amplicons? Please, clarify.

 

Results

Figure 1 in the pdf version has low-quality resolution, please double-check.

 

Discussion

Line 281: (Willinger, 2019) should be numbered.

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

The authors did not spend time for revising their manuscript at all. Moreover, the authors did not catch the point of comments. I think that it is difficult to publish this manuscript in the journal.

 

Please see the file attached.

Comments for author File: Comments.pdf