Antioxidant of the putat air (<i>barringtonia racemosa</i>) as a herbal remedy to improve sperm quality due to cigarette smoke in rats

Dahril Dahril; San Winata Badiri; Dasrul Dasrul; Vivi Keumala Mutiawati; Said Alfin Khalilullah

doi:10.12688/f1000research.127783.2

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Research Article

Revised

Antioxidant of the putat air (barringtonia racemosa) as a herbal remedy to improve sperm quality due to cigarette smoke in rats

[version 2; peer review: 1 approved, 1 not approved]

Dahril Dahril ¹, San Winata Badiri², Dasrul Dasrul³, Vivi Keumala Mutiawati⁴, Said Alfin Khalilullah ¹

Dahril Dahril ¹, San Winata Badiri², [...] Dasrul Dasrul³, Vivi Keumala Mutiawati⁴, Said Alfin Khalilullah ¹

PUBLISHED 22 Sep 2023

Author details Author details

¹ Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia
² Department of surgery, Universitas Syiah Kuala, Aceh, Indonesia
³ Reproductive Department, Universitas Syiah Kuala, Aceh, Indonesia
⁴ Clinical Pathology Department, Universitas Syiah Kuala, Aceh, Indonesia

Dahril Dahril
Roles: Conceptualization, Data Curation, Formal Analysis, Methodology, Resources, Supervision, Validation, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing

San Winata Badiri
Roles: Conceptualization, Data Curation, Supervision, Validation, Writing – Original Draft Preparation, Writing – Review & Editing

Dasrul Dasrul
Roles: Formal Analysis, Investigation, Validation, Writing – Original Draft Preparation

Vivi Keumala Mutiawati
Roles: Supervision, Validation

Said Alfin Khalilullah
Roles: Writing – Review & Editing

OPEN PEER REVIEW

REVIEWER STATUS

This article is included in the Plant Science gateway.

Abstract

Background: The objective of this study is to analyse the antioxidant in putat air (barringtonia racemosa) as an herbal to improve sperm quality due to cigarette smoke.
Methods: An experimental study was conducted at animal laboratory, Faculty Veteriner University of Syiah Kuala, Banda Aceh, Indonesia. A total of 30 male rats (Rattus norvegicus) were divided into 5 treatment groups. Group 1 as a negative control (N) were not exposed to cigarette smoke. Group 2 as a positive control (P) were exposed to cigarette smoke. Group 3 as treatment 1 (P1) were exposed to cigarette smoke and 100 mg/kg BW/day of the barringtonia racemosa seed extract. Group 4 as treatment 2 (P2) were exposed to cigarette smoke and given 150mg/kg BW/day of the barringtonia racemosa seed extract. Group 5 as treatment 3 (P3) were exposed to cigarette smoke and given 200 mg/kg BW/day of the barringtonia racemosa seed extract. The cigarette smoke exposure was carried out in a smoking chamber for a total of 1 hour/rat, and the barringtonia racemosa seed extract is given orally once a day for 30 days.
Results: The results of this study were analyzed by one-way ANOVA showed significant differences in the quality of the sperm group that received cigarette smoke exposure (p <0.05) compared to the administration of the barringtonia racemosa.
Improved sperm quality was seen better with the administration of barringtonia racemosa 150 kg/kg BW/day compared with 100 and 200 kg.
Conclusions: Antioxidants from barringtonia racemosa can improve sperm quality due to cigarette smoke.

Keywords

Barringtonia racemosa, seed extract, cigarette smoke, sperm quality

Corresponding authors: Dahril Dahril, Said Alfin Khalilullah

Competing interests: No competing interests were disclosed.

Grant information: The author(s) declared that no grants were involved in supporting this work.

Copyright: © 2023 Dahril D et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

How to cite: Dahril D, Badiri SW, Dasrul D et al. Antioxidant of the putat air (barringtonia racemosa) as a herbal remedy to improve sperm quality due to cigarette smoke in rats [version 2; peer review: 1 approved, 1 not approved]. F1000Research 2023, 12:55 (https://doi.org/10.12688/f1000research.127783.2) First published: 13 Jan 2023, 12:55 (https://doi.org/10.12688/f1000research.127783.1) Latest published: 01 Mar 2024, 12:55 (https://doi.org/10.12688/f1000research.127783.3)

Revised Amendments from Version 1

In version two of this article, we added several things, such as the reviewer's suggestion, which emphasized that Barringtonia racemosa is an antioxidant. Apart from that, in the methodology section we also explain the sperm analysis method.

See the authors' detailed response to the review by Oghenerobor B. Akpor
See the authors' detailed response to the review by Talukdar Dibyajyoti

Introduction

Male infertility is a problem that shows an increase the incidence in last decade. Lifestyle factors such as smoking have shown to be responsible for reducing infertility.¹^–³ Data from World Health Organization (WHO) in 2018, more than 2.9 billion of the world's population smoked and 80% lived in developing countries.⁴^–⁶ While the number of smokers in Indonesia reached 72 million population in 2015 and is predicted to increase to 83 million in 2020 with the proportion of men at 75.2% and the highest age group is 40-54 years.⁷ Around 100 toxic compounds such as tar, nicotine, nitrosamine, carbon monoxide, PAH compound (Polynuclear Aromatic Hydrogen), phenol, carbonyl, chlorindioxin, furan, and its smoke contains very high free radicals (ROS). These chemicals have the potential to damage the function of the testes and germ cells.⁸

Antioxidants have recently gained popularity due to their lower toxicity as compared to synthetic agents, also can naturally balance the scavenging system, and reduce intracellular ROS which will prevent decreased sperm quality due to substances from cigarette smoke.⁹ Antioxidant activity is essential as a research tool for evaluating the scientific basis for known traditional herbal treatments that claim to have antioxidant effects.

Barringtonia racemose, locally known as putat air in Indonesia, is a species of mangrove plant that grows in rice fields, originally originating from East Africa and Madagascar to Sri Lanka, India, and continuing into the Southeast Asian region, East Asia, northern Australia and the Micronesia and Polynesia Islands.¹⁰ As traditional remedy, this fruit have been used in the treatment of abscess, cough, asthma and diarrhea.¹¹ Several studies have succeeded in isolating some antioxidants from the leaves, kernels, fruit, roots, and stems of these plants such as polyphenols, flavonoids, and triterpenoids.¹⁰^,¹² The capacity of polyphenols, flavonoids, and triterpenoids as antioxidants is stronger than vitamins C and E.⁷

Different aerial sections of barringtonia racemosa have been found to exhibit strong antioxidant activity in earlier research.⁶^,⁷^,⁸^,¹¹ However, research on the edible shoots is lacking, notably on the antioxidant components and how various solvent extractions affect the antioxidant activities that arise. Our team's early assessment revealed that among 19 tropical plants, the shoots of barringtonia racemosa had one of the greatest antioxidant activity.¹⁰^,¹²

Despite a few isolated studies on certain parts of the plants, information on the antioxidant activity of the barringtonia racemose is still limited. The antioxidant properties of the plant extract may have given information on the mechanism relating to the plant's bioactive. This study evaluated the effect of the antioxidant in barringtonia racemosa an herbal to improving sperm quality due to cigarette smoke.

Methods

This study was registered to the Medical and Health Research Ethics Committee (362/EA/FK-RSUDZA/2019) and approved by the appropriate institutes’ ethics committee by Komite etik penelitian kesehatan Faculty of medicine, Universitas Syiah Kuala, Dr. Zainoel Abidin General Hospital. We collect data from 30 male rats (Rattus norvegicus) and were divided into five treatment groups. The period of study was 30 days. The interventions of cigarette smoke exposure were carried out in a smoking chamber for a total of one hour/rat for 30 days.

Chemical

All the chemicals and reagents used were of analytic grade and were obtained from the laboratory our institution. Baringtonia racemosa is obtained from rice fields in Aceh, Indonesia and then the seed extraction process is carried out in a laboratory. The male rats (Rattus norvegicus) in good health were also obtained from the same laboratory.

Tools

The tools used were rat cages, rat drinking places, smoking rooms, gastric sonde, gloves, small surgical instruments, knives, filter paper, incubators, vacuum rotary evaporators, water baths. Digital scales, glass objects, hot plates, cover glass, rotary microtomes, light microscopes, and cigarettes were also used.

Experimental animals and treatment

Male adult rats (Rattus norvegicus) strain of Wistar were healthy and aged 16-20 weeks from the animal laboratory of veterinary faculty with a bodyweight of 180-200 grams. All animals (n=30) were housed in rat cages and were fed on standard laboratory diet daily food and water ad libitum. Rats were kept on laboratory conditions, with standard temperature (25±2)°C, relative humidity of (40%–60%) and 12h/12h light and dark cycle.

The experimental rats were randomly divided into five groups, with 6 rats in each group. Group one as a negative control (N) in which the rats were not exposed to cigarette smoke. Group two-five received cigarette smoke exposure where group two as a positive control (P) and three-five were treated with barringtonia seed extract at doses of 100, 150, and 200 mg/kg BW/day orally.

Sample collection

In this study federer’s formula was used to calculate the sample size and the minumum sample is five each group. At the end of the experiment, the animals were sacrificed with cervical dislocation; both testicular regions along with the epididymis were dissected out to collect the sperm for further analysis. We carry out this procedure under animal anesthesia to reduce pain, suffering and distress. No unexpected adverse events were found during the experiment.

Sperm analysis

The quality of sperm were analyzed using the WHO 2010 method which included the amount, motility, viability and morphology.

Spermatozoa from the rat epididymis were aspirated using an erythrocyte pipette. Then, with the same pipette, 0.9% NaCl was sucked up to a limit of 101 so that 200 times the dilution results were obtained. Then, the mixture was discarded three drops and dropped into the Neubauer counting chamber. Observe and count the number of sperm under a light microscope with 400 times magnification using the formula N x 107 sperm/ml.

Statistical analysis

All data were analysed using the IBM SPSS 23.00 (IBM Corp., Chicago) and by using one-way analysis of variance (ANOVA). Shapiro–Wilk tests were used to analyze the normality of data. The data were expressed as mean ± standard deviation (mean±SD). Duncan test was applied for the comparison between the control group and each treated group individually. P<0.05 was considered statistically different.

Results

Phytochemical extract test

The phytochemical test results of the barringtonia racemosa can be seen in Table 1.

Table 1. Phytochemical test results of the ethyl acetate extract of barringtonia racemosa seed.

Metabolites content	Reagen of B. Racemosa kernel extract	Test Results	Observation Results
	Mayer	-	No white deposits formed
Alkaloids	Waner	-	No chocolate deposits formed
	Dragendorff	-	No red deposits formed
Steroid	Liebermann- Burchard Test	+	Formed bluish color
Terpenoid	Liebermann-Burchard Test	+	No red color formed
Saponin	Shuffle	+	Foaming
Flavonoid	HCl and Metal Mg	+	Reddish color formed
Phenolics	FeCl₃	+	Bluish black formed
Tanin	FeCl₃	+	Bluish black formed

In our experiment, it was found that the steroid compounds, terpenoid, saponins,flavonoids, phenolics, and tannins in barringtonia racemosa seed extract but not found alkaloids.

Sperm analysis

From results of sperm analysis after barringtonia racemosa seed extract administration for 30 days showed that sperm quality improved (Table 2).

Table 2. Sperm analysis after barringtonia racemosa seed extract administration.

Variable	N⁺	P^*	P1^**	P2^#	P3^##
Number of Sperma (x10⁶sp/ml)	65,00 ± 4,30^A	33,20 ± 4,92^D	40,60 ± 4,28^C	59,80 ± 3,35^A	50,80 ± 5,50^B
Sperm Motility (%)	77,40 ± 2,41^A	31,60 ± 3,97^E	42,00 ± 4,30^D	61,80 ± 6,22^B	50,60 ± 4,04^C
Viability of Sperm (%)	73,80 ± 4,32^A	34,20 ± 4,66^E	42,60 ± 3,13^D	61,00 ± 6,04^B	53,20 ± 3,96^C
Morphology of Sperm abnormal (%)	15,00 ± 3,16^D	60,20 ± 6,10^A	41,20 ± 7,19^B	28,60 ± 3,36^C	37,60 ± 5,41^B

+ (negative control) normal rats without exposure to cigarette

* (positive control) normal rats exposed to cigarette smoke

** normal rats exposed to cigarette smoke + 100 mg/kgBW barringtonia racemosa seed extract

# normal rats exposed to cigarette smoke + 150 mg/kgBW barringtonia racemosa seed extract

## normal rats exposed to cigarette smoke + 200 mg/kgBW barringtonia racemosa seed extract

Table 2 shows that sperm quality improved after administration of barringtonia racemosa seed extract in the treatment group (P1-3) compared with the positive control group (P) with statistical analysis (ANOVA) showing a significant difference (p <0.05). Further analysis with the Duncan test showed that administration of barringtonia racemosa seed extract at a dose of 150 mg/kgBW/day (P2) had a statistically significant improvement in sperm quality (P <0.05) compared with a dose of 100 mg/kg/day (P1) and 200 mg/kgBW/day (P3).

Discussion

This study shows a decrease in sperm quality in animal models due to exposure to cigarette smoke containing nicotine, tar, nitrosamine, carbon monoxide, phenol, carbonyl, chlorindioxin, and PAH compounds (Polynuclear Aromatic Hydrogen).²^–⁵

Nicotine is a toxic alkaloid found in tobacco smoke. Nicotine enters the brain quickly in about ten seconds. Nicotine can cross the brain barrier and circulate in all brain cells, then decrease rapidly, and circulate throughout the body within 15-20 minutes at the time of last exposure.¹³ Nicotine can stimulate the adrenal medulla to release catecholamines which can affect the central nervous system, thereby disrupting the mechanism of hormonal synthesis between the hypothalamus, anterior pituitary, and Leydig cells. Thus it will disrupt the process of spermatogenesis.¹⁴ Nicotine also interferes with cellular respiration processes in the mitochondria which leads to increased formation of reactive oxygen species (ROS) and induces the formation of oxidative stress by increasing pro-oxidants and reducing protection from antioxidants.¹³ Increased free radicals in the circulation will damage the spermatogenic cell membrane, DNA, spermatogenic cell growth, and disrupt the transport of important ions in proliferation thereby increasing sperm apoptosis and inhibiting spermatogenesis.¹⁵

One study reported that mice treated with cigarette smoke experienced DNA (Deoxiribo Nuclear Acid) fragmentation, decreased motility of the sperm, and reduced fertility rates.⁸ In previous studies conducted on animal experiments, it is known that cigarette smoke affects spermatogenesis in the seminiferous tubules and affects testosterone levels.⁸^,⁹

In this study, we found that the extract of barringtonia racemosa seed significantly improve sperm quality due to cigarette smoke (p < 0.05). this antioxidant can effectively protect body cells from oxidative stress including neutralizing free radicals caused by cigarette smoke and reducing sperm cell damage caused by ROS thereby avoiding a decrease in sperm quality.¹⁶ The barringtonia racemosa seed extract contains polyphenol compounds such as phenols, phenolic acids, flavonoids, and tannins that have potential as antioxidants. Polyphenol is an antioxidant carotenoid pigment that can bind a single oxidant ten times higher than vitamin C and one hundred times higher compared to α-tocopherol (vitamin E) and carotenoids.¹⁷^,¹⁸ Flavonoids are a group of polyphenol compounds as exogenous antioxidants that are useful in preventing cell damage due to oxidative stress, there are two mechanisms of flavonoids as antioxidants, which donate hydrogen ions to neutralize the toxic effects of free radicals and increase the expression of endogenous antioxidant genes through nuclear factor erythroid 2 related to factor 2 (Nrf2), increases genes that play a role in the synthesis of endogenous antioxidant enzymes.¹⁹ The strength of flavonoid antioxidant activity depends on the number and position of the -OH groups present in the molecule.²⁰ These phenolic antioxidants block the oxidation of lipids and other molecules by donating hydrogen atoms to radical compounds to form phenoxyl radical intermediates. The phenoxyl radical intermediate compounds are relatively stable so they are no longer able to initiate further radical reactions. High biological activity in these phenolic compounds lies in the position and number of the -OH group.²¹

Barringtonia racemosa seed extract also contains alkaloids, saponins, and triterpenoids which have activity in reducing MDA levels. Alkaloid compounds can act as scavengers of free radicals and prevent lipid peroxidation in microsomal hepatic preparation.²²^,²³ Triterpenoids can potentially induce Nrf2 gene expression and activate the ARE (Antioxidant Response Element) pathway in neuronal cells. Nrf2/ARE will regulate more than 200 genes including antioxidant genes.²⁴ Saponin compounds can act as antioxidants and have the ability to capture free radicals. The ability of saponin compounds can reduce oxidative stress in alloxan-induced mice.²¹

The highest sperm quality improvement was achieved at the concentration of barringtonia racemosa seed extract 150 mg/kg BW. This is caused by the difficulty of absorption of barringtonia racemosa seed extract with a higher concentration, so that the active ingredient of barringtonia racemosa seed extract is slow. On the other hand, decreased sperm quality in the treatment group of 200 mg/kg/day can also be caused by an increase in the active ingredient of barringtonia racemosa extract in the body. The high concentrations of antioxidant activity also will turn into prooxidants which can damage cells. When the dose of antioxidant and prooxidant is not balanced or high levels of oxidants while the low antioxidant, then the body will form prooxidant compounds to balance the levels with antioxidants.²⁵ This will increase the production of free radical cells. Prooxidant activity can reduce the level of antioxidant enzymes in the body such as glutathione peroxidase and catalase.²⁶ The imbalance between antioxidant enzymes and prooxidants in the body will damage spermatogenic cells, which will cause the formation of sperm to decrease.²⁷

Our study has several limitations due to the small sample size. However, the data's reliability can be maintained to avoid bias. Future studies with high-quality data is needed to evaluate this finding.

In conclusion, Antioxidants from barringtonia racemosa can improve sperm quality due to cigarette smoke. The Barringtonia racemosa extract at a dose of 150 mg/kgBW/day is better than the dose of 100 mg/BW/day and 200 mg/BW/day. This indicates that the optimum dose is 150 mg/BW/day.

Data availability

Underlying data

Figshare. Antioxidant of the Putat Air (Barringtonia racemosa) as a Herbal to Improve Sperm Quality due to Cigarette Smoke in rats. https://doi.org/10.6084/m9.figshare.21610341.v1.²⁸

Reporting guidelines

Figshare. ARRIVE guidelines checklist. DOI: https://doi.org/10.6084/m9.figshare.21746648.v1.²⁹

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Version 3

VERSION 3 PUBLISHED 13 Jan 2023

Author details Author details

San Winata Badiri
Roles: Conceptualization, Data Curation, Supervision, Validation, Writing – Original Draft Preparation, Writing – Review & Editing

Dasrul Dasrul
Roles: Formal Analysis, Investigation, Validation, Writing – Original Draft Preparation

Vivi Keumala Mutiawati
Roles: Supervision, Validation

Said Alfin Khalilullah
Roles: Writing – Review & Editing

Competing interests

No competing interests were disclosed.

Grant information

The author(s) declared that no grants were involved in supporting this work.

Article Versions (3)

version 3

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Published: 01 Mar 2024, 12:55

https://doi.org/10.12688/f1000research.127783.3

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© 2023 Dahril D et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Dahril D, Badiri SW, Dasrul D et al. Antioxidant of the putat air (barringtonia racemosa) as a herbal remedy to improve sperm quality due to cigarette smoke in rats [version 2; peer review: 1 approved, 1 not approved] F1000Research 2023, 12:55 (https://doi.org/10.12688/f1000research.127783.2)

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Reviewer Report 30 Nov 2023

Oghenerobor B. Akpor, Department of Biological Sciences, Afe Babalola University, Ado Ekiti, Ekiti, Nigeria

Not Approved

https://doi.org/10.5256/f1000research.156217.r208582

In the abstract, barringtonia racemosa should be correctly written as Barringtonia racemosa and italicised

The statement ‘The results of this study were analyzed by one-way ANOVA showed significant differences in the quality….’’ Should be part of methods and not result
Rattus norvegicus should be italicized and written as Rattus norvegicus

Generally, the abstract is written in an imbalanced format. There seems to be so much emphasis on the methods while result section is so scanty. The authors should address that

In my opinion, the authors did not make any significant amendment to the revised submission to justify a contrary decision.

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Plant growth promotion, biocomposites from agrowastes, bioactive properties of plants, microbial metabolites

I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above.

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Author Response 10 Apr 2024

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

10 Apr 2024

Author Response

Thank you reviewer for your suggestion to improve this manuscript. we already revision this new version manuscript according your comment especially in abstract, we already re written.
Competing Interests: No competing interests were disclosed
Thank you reviewer for your suggestion to improve this manuscript. we already revision this new version manuscript according your comment especially in abstract, we already re written.
Thank you reviewer for your suggestion to improve this manuscript. we already revision this new version manuscript according your comment especially in abstract, we already re written.
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Author Response 10 Apr 2024

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

10 Apr 2024

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Thank you reviewer for your suggestion to improve this manuscript. we already revision this new version manuscript according your comment especially in abstract, we already re written.
Competing Interests: No competing interests were disclosed
Thank you reviewer for your suggestion to improve this manuscript. we already revision this new version manuscript according your comment especially in abstract, we already re written.
Thank you reviewer for your suggestion to improve this manuscript. we already revision this new version manuscript according your comment especially in abstract, we already re written.
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Reviewer Report 10 Oct 2023

Talukdar Dibyajyoti, Central Agricultural University, Aizawl, Mizoram, India

Approved

https://doi.org/10.5256/f1000research.156217.r208581

This is an interesting study and the authors have collected a unique dataset. ... Continue reading

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Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

10 Apr 2024

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Thank you reviewer for the comment and approved this manuscript
Competing Interests: No competing interest
Thank you reviewer for the comment and approved this manuscript
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Author Response 10 Apr 2024

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

10 Apr 2024

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Thank you reviewer for the comment and approved this manuscript
Competing Interests: No competing interest
Thank you reviewer for the comment and approved this manuscript
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PUBLISHED 13 Jan 2023

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Reviewer Report 16 Aug 2023

Talukdar Dibyajyoti, Central Agricultural University, Aizawl, Mizoram, India

Approved with Reservations

https://doi.org/10.5256/f1000research.140324.r183392

Background of the study is not clear. Please rewrite.
Sperm analysis methodology needs to be rewritten with proper references.
Phytochemical: quantitative screening may be more appropriate.

Background of the study is not clear. Please rewrite.
Sperm analysis methodology needs to be rewritten with proper references.
Phytochemical: quantitative screening may be more appropriate.
The barringtonia racemosa; name should be italic in the text.
Some of sentences should be rephrase in the text.
In key word, please add the word rat
General comments: The aim of the study is to assess the antioxidant property of barringtonia racemosa which may be used as a herbal remedy to improve sperm quality due to cigarette smoke in rats. The present result of qualitative determination phytochemical compound of barringtonia racemosa does not claim the antioxidant property of barringtonia racemosa; quantitative screening need to be incorporated in the findings. The sperm analysis parameters i.e. sperm motility, viability, concentration and sperm abnormality are not rich enough to claim the possible improvement of sperm quality due to cigarette smoke in rats. The authors need to be include the plasma membrane integrity, acrosomal integrity, mitochondrial membrane integrity, DNA fragmentation of sperm cells, testosterone level, MDA level and if possible the sexual characteristics too.

Finally, I will not recommend the paper in the present form. The article needs to be modified and if possible the above mentioned parameters need to be incorporate.

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Partly
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

No

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Animal reproduction, Gyneacology, Obstetrics, Assisted Reproduction, Andrology

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.

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Author Response 16 Nov 2023

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

16 Nov 2023

Author Response

Thank you reviewer for your input. According to your input, we have made corrections to our manuscript.

Regarding your suggestion to add plasma membrane integrity, acrosomal integrity, mitochondrial membrane ... Continue reading Thank you reviewer for your input. According to your input, we have made corrections to our manuscript.

Regarding your suggestion to add plasma membrane integrity, acrosomal integrity, mitochondrial membrane integrity, DNA fragmentation of sperm cells, testosterone level and MDA level, we hope that we can continue this in our next study.
Thank you reviewer for your input. According to your input, we have made corrections to our manuscript.

Regarding your suggestion to add plasma membrane integrity, acrosomal integrity, mitochondrial membrane integrity, DNA fragmentation of sperm cells, testosterone level and MDA level, we hope that we can continue this in our next study.
Competing Interests: No competing Interest Close
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COMMENTS ON THIS REPORT

Author Response 16 Nov 2023

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

16 Nov 2023

Author Response

Thank you reviewer for your input. According to your input, we have made corrections to our manuscript.

Regarding your suggestion to add plasma membrane integrity, acrosomal integrity, mitochondrial membrane ... Continue reading Thank you reviewer for your input. According to your input, we have made corrections to our manuscript.

Regarding your suggestion to add plasma membrane integrity, acrosomal integrity, mitochondrial membrane integrity, DNA fragmentation of sperm cells, testosterone level and MDA level, we hope that we can continue this in our next study.
Thank you reviewer for your input. According to your input, we have made corrections to our manuscript.

Regarding your suggestion to add plasma membrane integrity, acrosomal integrity, mitochondrial membrane integrity, DNA fragmentation of sperm cells, testosterone level and MDA level, we hope that we can continue this in our next study.
Competing Interests: No competing Interest Close
Report a concern

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Reviewer Report 23 May 2023

Oghenerobor B. Akpor, Department of Biological Sciences, Afe Babalola University, Ado Ekiti, Ekiti, Nigeria

Olakunle Afolabi, Chemical Sciences, Afe Babalola University, Ado Ekiti, Ekiti, Nigeria

Not Approved

https://doi.org/10.5256/f1000research.140324.r171633

Background:
What is indicated in the background more of an aim/objective. There is no indication of background at all.

Methods
Tools: The authors do not need to mention the materials used. They should naturally reflect as the procedures are explained

Sample collection: Show how the Federer's formula was applied to arrive at five animals per grouping

Phytochemical: Qualitative screening alone is not adequate for the study. It would have been more ideal to carry out quantitative screening

General comments/observation
Based on the title of the paper, the work was aimed at assessing the antioxidant and sperm quality improvement potential of Barrintonia racemosa. Based on the results of the study, there is no indication of antioxidant but just qualitative determination of phytochemical components. There was no study on antioxidant properties/potential.

Besides, the data on sperm quality are not rich enough to justify a possible improvement in sperm quality. The authors will need to include histomorphological data. Data on testosterone profile and Malondialdehyde (MDA), which is a vital indicator of oxidative stress is also vital. In addition, data on luteinizing hormone (LH) levels, which in important in sexual development and functioning should be provided.

I will not recommend indexing of the paper in the present form. The data generated is extremely too scant to make the claim.

Is the work clearly and accurately presented and does it cite the current literature?

No
Is the study design appropriate and is the work technically sound?

No
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

Yes
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

No

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Plant growth promotion, bioactive properties of plants, microbial metabolites

We confirm that we have read this submission and believe that we have an appropriate level of expertise to state that we do not consider it to be of an acceptable scientific standard, for reasons outlined above.

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Author Response 16 Nov 2023

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

16 Nov 2023

Author Response

Dear Oghenerobor B. Akpor.

Thank you for your reviewers our manuscript.

We are very grateful for your input for the improvement of our manuscript. but related to that, we ... Continue reading Dear Oghenerobor B. Akpor.

Thank you for your reviewers our manuscript.

We are very grateful for your input for the improvement of our manuscript. but related to that, we need to convey a number of things that Barringtonia racemose, locally known as putat air in Indonesia, is an anti-oxidant, as we have stated in the introduction.

Besides that, additional examinations as you suggest such as histomorphological data, testosterone level, LH level and others are very informative data and can provide more information not only about sperm quality but also more complete sexual quality. We really hope that this preliminary study can continue with evaluation of more complex variables such as additional data as you mentioned above.
Dear Oghenerobor B. Akpor.

Thank you for your reviewers our manuscript.

We are very grateful for your input for the improvement of our manuscript. but related to that, we need to convey a number of things that Barringtonia racemose, locally known as putat air in Indonesia, is an anti-oxidant, as we have stated in the introduction.

Besides that, additional examinations as you suggest such as histomorphological data, testosterone level, LH level and others are very informative data and can provide more information not only about sperm quality but also more complete sexual quality. We really hope that this preliminary study can continue with evaluation of more complex variables such as additional data as you mentioned above.
Competing Interests: No competing interests were disclosed Close
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COMMENTS ON THIS REPORT

Author Response 16 Nov 2023

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

16 Nov 2023

Author Response

Dear Oghenerobor B. Akpor.

Thank you for your reviewers our manuscript.

We are very grateful for your input for the improvement of our manuscript. but related to that, we ... Continue reading Dear Oghenerobor B. Akpor.

Thank you for your reviewers our manuscript.

We are very grateful for your input for the improvement of our manuscript. but related to that, we need to convey a number of things that Barringtonia racemose, locally known as putat air in Indonesia, is an anti-oxidant, as we have stated in the introduction.

Besides that, additional examinations as you suggest such as histomorphological data, testosterone level, LH level and others are very informative data and can provide more information not only about sperm quality but also more complete sexual quality. We really hope that this preliminary study can continue with evaluation of more complex variables such as additional data as you mentioned above.
Dear Oghenerobor B. Akpor.

Thank you for your reviewers our manuscript.

We are very grateful for your input for the improvement of our manuscript. but related to that, we need to convey a number of things that Barringtonia racemose, locally known as putat air in Indonesia, is an anti-oxidant, as we have stated in the introduction.

Besides that, additional examinations as you suggest such as histomorphological data, testosterone level, LH level and others are very informative data and can provide more information not only about sperm quality but also more complete sexual quality. We really hope that this preliminary study can continue with evaluation of more complex variables such as additional data as you mentioned above.
Competing Interests: No competing interests were disclosed Close
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Version 3

VERSION 3 PUBLISHED 13 Jan 2023

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	1	2
Version 3 (revision) 01 Mar 24
Version 2 (revision) 22 Sep 23	read	read
Version 1 13 Jan 23	read	read

Oghenerobor B. Akpor, Afe Babalola University, Ado Ekiti, Nigeria

Olakunle Afolabi, Afe Babalola University, Ado Ekiti, Nigeria
Talukdar Dibyajyoti, Central Agricultural University, Aizawl, India

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Reviewer Report

6 Views

30 Nov 2023 | for Version 2

Oghenerobor B. Akpor, Department of Biological Sciences, Afe Babalola University, Ado Ekiti, Ekiti, Nigeria

6 Views Cite this report Responses(1)

Not Approved

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Plant growth promotion, biocomposites from agrowastes, bioactive properties of plants, microbial metabolites

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Author Response

10 Apr 2024

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

Thank you reviewer for your suggestion to improve this manuscript. we already revision this new version manuscript according your comment especially in abstract, we already re written.

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Competing Interests

No competing interests were disclosed

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Reviewer Report

10 Views

10 Oct 2023 | for Version 2

Talukdar Dibyajyoti, Central Agricultural University, Aizawl, Mizoram, India

10 Views Cite this report Responses(1)

Approved

This is an interesting study and the authors have collected a unique dataset. The paper is well written and structured. It can be accepted for indexing.

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Animal reproduction, Gyneacology, Obstetrics, Assisted Reproduction, Andrology

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

Respond to this report

Responses (1)

Author Response

10 Apr 2024

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

Thank you reviewer for the comment and approved this manuscript

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No competing interest

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Reviewer Report

14 Views

16 Aug 2023 | for Version 1

Talukdar Dibyajyoti, Central Agricultural University, Aizawl, Mizoram, India

14 Views Cite this report Responses(1)

Approved With Reservations

Background of the study is not clear. Please rewrite.
Sperm analysis methodology needs to be rewritten with proper references.
Phytochemical: quantitative screening may be more appropriate.
The barringtonia racemosa; name should be italic in the text.
Some of sentences should be rephrase in the text.
In key word, please add the word rat
General comments: The aim of the study is to assess the antioxidant property of barringtonia racemosa which may be used as a herbal remedy to improve sperm quality due to cigarette smoke in rats. The present result of qualitative determination phytochemical compound of barringtonia racemosa does not claim the antioxidant property of barringtonia racemosa; quantitative screening need to be incorporated in the findings. The sperm analysis parameters i.e. sperm motility, viability, concentration and sperm abnormality are not rich enough to claim the possible improvement of sperm quality due to cigarette smoke in rats. The authors need to be include the plasma membrane integrity, acrosomal integrity, mitochondrial membrane integrity, DNA fragmentation of sperm cells, testosterone level, MDA level and if possible the sexual characteristics too.

Finally, I will not recommend the paper in the present form. The article needs to be modified and if possible the above mentioned parameters need to be incorporate.

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Partly
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

No

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Animal reproduction, Gyneacology, Obstetrics, Assisted Reproduction, Andrology

Respond to this report

Responses (1)

Author Response

16 Nov 2023

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

Thank you reviewer for your input. According to your input, we have made corrections to our manuscript.

Regarding your suggestion to add plasma membrane integrity, acrosomal integrity, mitochondrial membrane integrity, DNA fragmentation of sperm cells, testosterone level and MDA level, we hope that we can continue this in our next study.

View more View less

Competing Interests

No competing Interest

Back to all reports

Reviewer Report

17 Views

23 May 2023 | for Version 1

Oghenerobor B. Akpor, Department of Biological Sciences, Afe Babalola University, Ado Ekiti, Ekiti, Nigeria

Olakunle Afolabi, Chemical Sciences, Afe Babalola University, Ado Ekiti, Ekiti, Nigeria

17 Views Cite this report Responses(1)

Not Approved

Is the work clearly and accurately presented and does it cite the current literature?

No
Is the study design appropriate and is the work technically sound?

No
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

Yes
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

No

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Plant growth promotion, bioactive properties of plants, microbial metabolites

Respond to this report

Responses (1)

Author Response

16 Nov 2023

Dahril Dahril, Department of Surgery, Division of Urologi, Universitas Syiah Kuala, Aceh, Indonesia

Dear Oghenerobor B. Akpor.

Thank you for your reviewers our manuscript.

We are very grateful for your input for the improvement of our manuscript. but related to that, we need to convey a number of things that Barringtonia racemose, locally known as putat air in Indonesia, is an anti-oxidant, as we have stated in the introduction.

Besides that, additional examinations as you suggest such as histomorphological data, testosterone level, LH level and others are very informative data and can provide more information not only about sperm quality but also more complete sexual quality. We really hope that this preliminary study can continue with evaluation of more complex variables such as additional data as you mentioned above.

View more View less

Competing Interests

No competing interests were disclosed

Alongside their report, reviewers assign a status to the article:

Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested

Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.

Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions

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[2] 2. Colagar A, Jorsaraee G, Marzony E: Cigarette smoking and the risk of male infertility. Biol Sci. 2007; 1.

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Antioxidant of the putat air (barringtonia racemosa) as a herbal remedy to improve sperm quality due to cigarette smoke in rats

Abstract

Keywords

Revised Amendments from Version 1

Introduction

Methods

Chemical

Tools

Experimental animals and treatment

Sample collection

Sperm analysis

Statistical analysis

Results

Phytochemical extract test

Table 1. Phytochemical test results of the ethyl acetate extract of barringtonia racemosa seed.

Sperm analysis

Table 2. Sperm analysis after barringtonia racemosa seed extract administration.

Discussion

Data availability

Underlying data

Reporting guidelines

References

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