Antimicrobial activity of Terminalia catappa brown leaf extracts against Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853

Ovin Qonita Allyn; Eko Kusumawati; Rudy Agung Nugroho

doi:10.12688/f1000research.15998.1

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Antimicrobial activity of Terminalia catappa brown leaf extracts against Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853

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

Ovin Qonita Allyn¹, Eko Kusumawati², Rudy Agung Nugroho ¹

PUBLISHED 04 Sep 2018

Author details Author details

¹ Animal Physiology, Development, and Molecular Laboratory, Department of Biology, Faculty of Mathematics and Natural Sciences, Mulawarman University, Samarinda, Kalimantan Timur, 75123, Indonesia
² Microbiology and Genetic Molecular Laboratory, Department of Biology, Faculty of Mathematics and Natural Sciences, Mulawarman University, Samarinda, Kalimantan Timur, 75123, Indonesia

Ovin Qonita Allyn
Roles: Conceptualization, Formal Analysis, Investigation, Methodology, Resources, Validation

Eko Kusumawati
Roles: Methodology, Supervision, Writing – Original Draft Preparation

Rudy Agung Nugroho
Roles: Data Curation, Investigation, Supervision, Validation, Writing – Review & Editing

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REVIEWER STATUS

Abstract

The aim of this study was to determine the effects of various concentration of Terminalia catappa brown leaves extract which can inhibit the growth of Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853. The crushed-brown leaves of Terminalia catappa was extracted using 95% ethanol, filtered, and evaporated. The dried T. catappa extract was used to identify phytochemical content qualitatively. Total phenolic and flavonoid contents were also measured quantitatively from dried extract. The dried extracts were also dissolved in sterile aquadest and serial dilutions were prepared to final concentration of 30, 60 and 90%. A disc diffusion method was used to evaluate the antibacterial activity of various concentrations of ethanol extract of brown leaves of T. catappa. Inhibition zone diameter was measured to determine antibacterial activity. Gentamycin sulfate and distilled water were used as positive and negative controls, respectively. Dried ethanolic extract of brown T. catappa leaves contained flavonoid, quinon, phenolic, triterpenoid, and tannin. A total of 208.722 mg gallic acid equivalent/g extract of total phenolic and 35.7671 mg quercetin equivalent/g extract of total flavonoid were also found in the dried extract. The inhibition zone diameters of ethanolic extracts ranged from 1.73 to 9.06 mm (S. aureus) and from 1.83 to 6.5 mm (P. aeruginosa). The higher concentration of extract, the wider the inhibition zone diameters for both bacteria. P. aeruginosa was more resistant to high concentrations of extract (90%) than S. aureus. Ethanolic extracts of the brown leaves of T. catappa had different antibacterial effects against S. aureus and P. aeruginosa. The higher the concentration of extract, the wider the inhibition zone diameter for both bacteria. P. aeruginosa was more resistant to high concentrations of ethanolic extracts of the brown leaves of T. catappa.

Keywords

Terminalia catappa, phytochemicals, Staphylococcus aureus, Pseudomonas aeruginosa, antibacterial

Corresponding author: Rudy Agung Nugroho

Competing interests: No competing interests were disclosed.

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

Copyright: © 2018 Allyn OQ 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. Data associated with the article are available under the terms of the Creative Commons Zero "No rights reserved" data waiver (CC0 1.0 Public domain dedication).

How to cite: Allyn OQ, Kusumawati E and Nugroho RA. Antimicrobial activity of Terminalia catappa brown leaf extracts against Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853 [version 1; peer review: 2 approved, 1 not approved]. F1000Research 2018, 7:1406 (https://doi.org/10.12688/f1000research.15998.1) First published: 04 Sep 2018, 7:1406 (https://doi.org/10.12688/f1000research.15998.1) Latest published: 04 Sep 2018, 7:1406 (https://doi.org/10.12688/f1000research.15998.1)

Introduction

Staphylococcus and Pseudomonas species have been identified as causative agents of disease and serious pathogens in many aquatic animals, including fish^1–4, resulting in high mortality rates in many commercially farmed fish. Among various Staphylococcus and Pseudomonas species, Staphylococcus aureus and Pseudomonas aeruginosa are known to cause disease in Oreochromis niloticus and Oreochromis mossambicus⁵. To reduce high mortality rates in farmed fish, aquaculturists and researcher used chemical agents and antibiotics to promote growth or prevent S. aureus and P. aeruginosa infection⁶.

However, the use of antibiotics to prevent and cure common infectious diseases in fish is becoming increasingly limited due to environmental concern, and increasingly expensive and ineffective because of microbial resistance^7–9. As alternatives, various plant extracts, such as those of Boesenbergia pandurata, Zingiber zerumbet and Solanum ferox, have been tested and used as an alternative to antibiotics^10–12. Another potential plant extract that can be used as an antimicrobial is that of Terminalia catappa, which is widely distributed in tropical and sub-tropical regions, including Indonesia^13,14.

Terminalia catappa L., belonging to the family Combretaceae, is a large deciduous tree. The aqueous extract of Terminalia catappa leaves has been known as a folk medicine for antipyretic, hemostatic, hepatitis and liver-related diseases purposes in the Philippines, Malaysia and Indonesia^15,16. Past research revealed that the extract of T. catappa leaves can be used to improve a resistance to Aeromonas hydrophila in Betta sp¹⁷, remedy against tilapia (Oreochromis niloticus) parasites and bacterial pathogen^18,19. Nevertheless, scientific literature concerning the antibacterial potency of T. catappa against Staphylococcus aureus and Pseudomonas aeruginosa is limited.

Thus, the aim of the study was to evaluate the effects of various concentration of T. catappa brown leaf extract on the growth of Staphylococcus aureus and Pseudomonas aeruginosa by calculating inhibition zone diameters. The phytochemical content of the extract was also qualitatively determined and the flavonoid and phenolic concentrations in the extract was quantified.

Methods

Site and time

The research was performed from March to May 2018 at the Animal Physiology, Development and Molecular Laboratory for extracting T. catappa leaves. Meanwhile, assay study was done at microbiology and molecular genetic laboratory.

Bacterial strains and culture condition

Bacterial strains were obtained from a Microbiology Laboratory, Faculty of Pharmacy, Sumatera Utara University, Indonesia. Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853 were used to investigate the antibacterial activity. Both bacteria were sub-cultured on nutrient agar and stored at 4°C until use.

Plant materials

Brown leaves of T. catappa were collected from a region of Mulawarman university campus, Samarinda, East Kalimantan. Leaves were dried at room temperature for 2 days, crushed, transferred into a glass container and preserved until the extraction procedure.

Extraction procedure

Approximately 1 kg of crushed leaves was soaked in 1 l of 95% ethanol for 5 days and shaken occasionally with a shaker. After 5 days, materials were filtered (Whatman No. 11 paper filter). The filtrate was evaporated using a rotary evaporator. Finally, the dried extracts were obtained and stored at 4°C in a dark bottle until use. The dried extracts were then dissolved in sterile distilled water and serial dilutions were prepared to give final concentrations of 30, 60 and 90%.

Phytochemical content

Dried extract samples were subjected to qualitative phytochemical analysis for flavonoids, quinon, alkaloids, phenolic, steroid, triterpenoid, saponins, and tannins using standard methods as previously described by Nugroho et al.²⁰. Meanwhile, total phenolics and flavonoids were quantitatively measured, using the method described by Pourmorad et al.²¹.

Antibacterial activity assay

The antibacterial activity of T. catappa brown leaf ethanolic extract was evaluated using the disc diffusion method²². Three replicated agar plates were used for each different concentration and both controls (distilled water and 0.1% gentamycin sulfate). A total of 10 µl extract was added to a paper disc for each concentration and controls. Each disk was then placed in agar plate which had bacterial suspension in the plates. All plates were incubated at 37°C for 24 h. The diameter of inhibition zone created by each disc was measured (in mm) using a micrometer.

Data analysis

The inhibition zone data were expressed as means ± standard error. The data were subjected to ANOVA, followed by Duncan’s post hoc test to evaluate significant differences among the groups of treatments. Meanwhile, the comparison between bacteria in each concentration was performed using a t-test. All significant tests were at P<0.05 levels and all analysis was done using SPSS 22 (SPSS, Inc., USA). The data of the phytochemical content and the concentration of flavonoid and phenolic were analyzed descriptively.

Results and discussion

The dried extract of T. catappa brown leaves contained flavonoids, quinon, phenolics, triterpenoids, and tannins. There were no alkaloids, steroids or saponin found in the dried extract. Total phenolic (208.722 mg gallic acid equivalent/g extract) and total flavonoid (35.7671 mg quercetin equivalent/g extract) were detected in the dried extract. The inhibition zone diameters of ethanolic extracts ranged from 1.73 to 9.06 mm for S. aureus, and from 1.83 to 6.5 mm for P. aeruginosa. Increasing the extract concentration increased the inhibition zone diameters for both bacteria (Figure 1). P. aeruginosa was more resistant to high concentrations of extract (90%) than S. aureus (Table 1). According to Xie et al.²³, flavonoids are known antibacterial agents against a wide range of pathogenic bacteria. In addition, Fu et al.²⁴ also revealed that phenolic extracts from some plants also have antibacterial effects against many kinds of bacteria. The data showing the inhibition zone diameters for both bacteria at each concentration of extract can be seen in Dataset 1²⁵.

Dataset 1.Inhibition zone diameters for both bacteria at different concentration of extracts and images of every repeat experiment performed.

Figure 1. Inhibition zone of Terminalia catappa brown leaves ethanolic extract against Staphylococcus aureus and Pseudomonas aeruginosa.

(a) Negative control, (b) positive control (0.1% gentamycin sulfate), Terminalia catappa extract (c) 30%, (d) 60%, (e) 90%. Images shown are representative of n=3 repeats.

Table 1. Inhibitory zone diameter (in mm) of Staphylococcus aureus and Pseudomonas aeruginosa after treated with different concentration of brown leaves of T. catappa ethanolic extract.

Bacteria	Positive control	Extract concentration
Bacteria	Positive control	30%	60%	90%
Staphylococcus aureus	5.78±0,27^a,1	1.73±0,24^b,1	5.28±1,06^a,c,1	9.06±0,56^d,1
Pseudomonas aeruginosa	7.15±0,20^a,2	1.83±0,87^b,1	4.53±0,78^c,1	6.50±0,13^a,d,2

Different superscript letters in the same row indicate significantly different mean values for different treatments at P<0.05. Different superscript numbers in the same column indicate significantly different mean values for different treatments at P<0.05. The negative control was omitted as no inhibition zone was present. Positive control, 0.1% gentamycin sulfate.

Conclusion

Ethanolic extracts of the brown leaves of T. catappa have potential antibacterial effects against S. aureus and P. aeruginosa, indicated by the inhibition zone formed around the extract. The inhibition zone diameter increased with increasing concentrations of T. catappa extract. P. aeruginosa exhibited more resistance to high concentrations of ethanol extracts of the brown leaves of T. catappa than S. aureus.

Data availability

Dataset 1. Inhibition zone diameters for both bacteria at different concentration of extracts and images of every repeat experiment performed. DOI: https://doi.org/10.5256/f1000research.15998.d215169²⁵.

Grant information

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

Acknowledgments

The authors thank to Faculty of Mathematics and Natural Sciences, Mulawarman University, Samarinda, East Kalimantan. The appreciation goes to all of our students who helped the authors during the trial.

Faculty Opinions recommended

References

1. Najiah M, Aqilah NI, Lee KL, et al.: Massive mortality associated with Streptococcus agalactiae infection in cage-cultured red hybrid tilapia Oreochromis niloticus in Como River, Kenyir Lake, Malaysia. J Biol Sci. 2012; 12(8): 438–442. Publisher Full Text
2. Na-Phatthalung P, Chusri S, Suanyuk N, et al.: In vitro and in vivo assessments of Rhodomyrtus tomentosa leaf extract as an alternative anti-streptococcal agent in Nile tilapia (Oreochromis niloticus L.). J Med Microbiol. 2017; 66(4): 430–439. PubMed Abstract | Publisher Full Text
3. Saikia DJ, Chattopadhyay P, Banerjee G, et al.: Time and dose dependent effect of Pseudomonas aeruginosa infection on the scales of Channa punctata (Bloch) through light and electron microscopy. Turk J Fish Aquat Sci. 2017; 17(5): 871–876. Publisher Full Text
4. Baldissera MD, Souza CF, Santos RCV, et al.: Pseudomonas aeruginosa strain PA01 impairs enzymes of the phosphotransfer network in the gills of Rhamdia quelen. Vet Microbiol. 2017; 201: 121–125. PubMed Abstract | Publisher Full Text
5. Thomas J, Thanigaivel S, Vijayakumar S, et al.: Pathogenecity of Pseudomonas aeruginosa in Oreochromis mossambicus and treatment using lime oil nanoemulsion. Colloids Surf B Biointerfaces. 2014; 116: 372–377. PubMed Abstract | Publisher Full Text
6. Grema HA, Geidam YA, Suleiman A, et al.: Multi-Drug Resistant Bacteria Isolated from Fish and Fish Handlers in Maiduguri, Nigeria. Int J Anim Vet Adv. 2015; 7(3): 49–54. Reference Source
7. Samanidou VF, Evaggelopoulou EN: Analytical strategies to determine antibiotic residues in fish. J Sep Sci. 2007; 30(16): 2549–2569. PubMed Abstract | Publisher Full Text
8. Uchida K, Konishi Y, Harada K, et al.: Monitoring of Antibiotic Residues in Aquatic Products in Urban and Rural Areas of Vietnam. J Agric Food Chem. 2016; 64(31): 6133–8. PubMed Abstract | Publisher Full Text
9. He X, Deng M, Wang Q, et al.: Residues and health risk assessment of quinolones and sulfonamides in cultured fish from Pearl River Delta, China. Aquacult. 2016; 458: 38–46. Publisher Full Text
10. Hardi EH, Kusuma IW, Suwinarti W, et al.: Antibacterial activities of some Borneo plant extracts against pathogenic bacteria of Aeromonas hydrophila and Pseudomonas sp. AACL Bioflux. 2016; 9(3): 638–646. Reference Source
11. Hardi EH, Kusuma IW, Suwinarti W, et al.: Short Communication: Antibacterial activity of Boesenbergia pandurata, Zingiber zerumbet and Solanum ferox extracts against Aeromonas hydrophila and Pseudomonas sp. Nusantara Bioscience. 2016; 8(1): 18–21. Publisher Full Text
12. Hardi EH, Saptiani G, Kusuma IW, et al.: Immunomodulatory and antibacterial effects of Boesenbergia pandurata, Solanum ferox, and Zingiber zerumbet on tilapia, Oreochromis niloticus. AACL/Bioflux. 2017; 10(2): 182–190. Reference Source
13. Hyttel P, Sinowatz F, Vejlsted M: Essentials of Domestic Animal Embryology. Saunders/Elsevier. 2010. Reference Source
14. Hyttel P, Sinowatz F, Vejlsted M, et al.: Essentials of domestic animal embryology. Elsevier Health Sciences UK. 2009. Reference Source
15. Meena K, Raja TK: Immobilization of Saccharomyces cerevisiae cells by gel entrapment using various metal alginates. World J Microbiol Biotechnol. 2006; 22(6): 651–653. Publisher Full Text
16. Vučurović VM, Razmovski RN: Sugar beet pulp as support for Saccharomyces cerivisiae immobilization in bioethanol production. Ind Crops Prod. 2012; 39: 128–134. Publisher Full Text
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18. Goh CS, Tan KT, Lee KT, et al.: Bio-ethanol from lignocellulose: Status, perspectives and challenges in Malaysia. Bioresour Technol. 2010; 101(13): 4834–41. PubMed Abstract | Publisher Full Text
19. Öztop HN, Öztop AY, Işikver Y, et al.: Immobilization of Saccharomyces cerevisiae on to radiation crosslinked HEMA/AAm hydrogels for production of ethyl alcohol. Process Biochem. 2002; 37(6): 651–657. Publisher Full Text
20. Nugroho RA, Manurung H, Saraswati D, et al.: The effects of Terminalia catappa leaf extract on the haematological profile of ornamental fish Betta splendens. Biosaintifika: Journal of Biology and Biology Education. 2016; 8(2): 241–248.
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22. Reddy PS, John MS, Devi PV, et al.: Detection of vancomycin susceptibility among clinical isolates of MRSA by using minimum inhibitory concentration method. Int J Res Med Sci. 2015; 3(6): 1378–1382. Publisher Full Text
23. Xie Y, Yang W, Tang F, et al.: Antibacterial activities of flavonoids: structure-activity relationship and mechanism. Curr Med Chem. 2015; 22(1): 132–49. PubMed Abstract | Publisher Full Text
24. Fu L, Lu W, Zhou X: Phenolic Compounds and In Vitro Antibacterial and Antioxidant Activities of Three Tropic Fruits: Persimmon, Guava, and Sweetsop. Biomed Res Int. 2016; 2016: 4287461. PubMed Abstract | Publisher Full Text | Free Full Text
25. Allyn OQ, Kusumawati E, Nugroho RA: Dataset 1 in: Antimicrobial activity of Terminalia catappa brown leaf extracts against Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853. F1000Research. 2018. http://www.doi.org/10.5256/f1000research.15998.d215169

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Version 1

VERSION 1 PUBLISHED 04 Sep 2018

Author details Author details

¹ Animal Physiology, Development, and Molecular Laboratory, Department of Biology, Faculty of Mathematics and Natural Sciences, Mulawarman University, Samarinda, Kalimantan Timur, 75123, Indonesia
² Microbiology and Genetic Molecular Laboratory, Department of Biology, Faculty of Mathematics and Natural Sciences, Mulawarman University, Samarinda, Kalimantan Timur, 75123, Indonesia

Ovin Qonita Allyn
Roles: Conceptualization, Formal Analysis, Investigation, Methodology, Resources, Validation

Eko Kusumawati
Roles: Methodology, Supervision, Writing – Original Draft Preparation

Rudy Agung Nugroho
Roles: Data Curation, Investigation, Supervision, Validation, 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 (1)

version 1

Published: 04 Sep 2018, 7:1406

https://doi.org/10.12688/f1000research.15998.1

Copyright

© 2018 Allyn OQ 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. Data associated with the article are available under the terms of the Creative Commons Zero "No rights reserved" data waiver (CC0 1.0 Public domain dedication).

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Allyn OQ, Kusumawati E and Nugroho RA. Antimicrobial activity of Terminalia catappa brown leaf extracts against Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853 [version 1; peer review: 2 approved, 1 not approved] F1000Research 2018, 7:1406 (https://doi.org/10.12688/f1000research.15998.1)

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

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Version 1

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Reviewer Report 23 Oct 2018

Cimi Ilmiawati, Division of Environmental Toxicology, Department of Pharmacology, Faculty of Medicine, Andalas University, Padang, Indonesia

Approved

https://doi.org/10.5256/f1000research.17473.r37962

Comments:

This study contributes information on the potential of T. catappa leaves extract as an antibacterial alternative to address antibacterial resistance problems in fish farming.

Methods
Site and time: Please specify the affiliation ... Continue reading

Comments:

This study contributes information on the potential of T. catappa leaves extract as an antibacterial alternative to address antibacterial resistance problems in fish farming.

Methods
Site and time: Please specify the affiliation of the laboratory (whether it belongs to a department in a university or else).

Plant materials
Please correct the writing of Mulawarman University.
How to preserve the crushed leaves? How long the preservation until the extraction?

Phytochemical contents
What is the purpose of qualitative and quantitave phytochemical analysis in relation to the study objectives? Please elaborate in the introduction part.

Results

The inhibition zone in Figure 1 for the positive control is not very clear, therefore a zoom in figure is required to judge the result.
I was expecting to see three discs in each plate of treatment and this experiment shall be replicated three times, before analyzing the data and arriving into a conclusion. Please comment on your replication method.

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Pharmacology, toxicology, molecular endocrinology

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.

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Reviewer Report 23 Oct 2018

Md Fakruddin, Laboratory of stem cell stress, International Research Center for Medical Sciences, Kumamoto University, Kumamoto , Japan

Not Approved

https://doi.org/10.5256/f1000research.17473.r37961

In the title, it should be mention ethanol extract, as the authors only evaluate ethanol extract of the leaves.
In the figure 1, the quality is poor, so it seems the positive control do not have

In the title, it should be mention ethanol extract, as the authors only evaluate ethanol extract of the leaves.
In the figure 1, the quality is poor, so it seems the positive control do not have any inhibition zone!
Only one strain of each bacteria was tested, it would be more authentic if more types of strains as well as some wild or environmental strains were tested.
MIC/MBC of the extract against these bacteria should be determined.
Data of this study should be compared with similar data published already. It is understood, that there may be no data of this particular species, but other species of terminalia can be consulted.

Competing Interests: No competing interests were disclosed.

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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Reviewer Report 20 Sep 2018

Edwin Setiawan, Department of Biology, Faculty of Natural Sciences, Sepuluh Nopember Institute of Technology, Surabaya, Indonesia

Approved

https://doi.org/10.5256/f1000research.17473.r37963

The study is designed appropriately and and technical procedure has been taken sufficiently to answer the research question. Furthermore, methods and analyses process also sufficient to be replicated by readers. In addition, statistical method that used is also appropriate. They ... Continue reading

The study is designed appropriately and and technical procedure has been taken sufficiently to answer the research question. Furthermore, methods and analyses process also sufficient to be replicated by readers. In addition, statistical method that used is also appropriate. They author need to highlighted and emphasized on the results and discussion with expanding their discussion with comparing another result that similar to this study.

In the result and discussion part, it is better to compare phenolic and flavonoid content of Terminalia catappa to other plant extract that close to Terminalia catappa if possible. Or in other words, comparing other similar research that used plant bioactive compound for antibacterial to research. Therefore, this result and discussion part could be expanded comprehensively.

In addition, simple and short explanation on how the mechanism flavonoid and phenolic inhibit bacterial growth can be added in this part.

Competing Interests: No competing interests were disclosed.

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.

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Edwin Setiawan, Sepuluh Nopember Institute of Technology, Surabaya, Indonesia
Md Fakruddin, International Research Center for Medical Sciences, Kumamoto University, Kumamoto , Japan
Cimi Ilmiawati, Andalas University, Padang, Indonesia

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23 Oct 2018 | for Version 1

Cimi Ilmiawati, Division of Environmental Toxicology, Department of Pharmacology, Faculty of Medicine, Andalas University, Padang, Indonesia

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Approved

Comments:

This study contributes information on the potential of T. catappa leaves extract as an antibacterial alternative to address antibacterial resistance problems in fish farming.

Methods
Site and time: Please specify the affiliation of the laboratory (whether it belongs to a department in a university or else).

Plant materials
Please correct the writing of Mulawarman University.
How to preserve the crushed leaves? How long the preservation until the extraction?

Phytochemical contents
What is the purpose of qualitative and quantitave phytochemical analysis in relation to the study objectives? Please elaborate in the introduction part.

Results

The inhibition zone in Figure 1 for the positive control is not very clear, therefore a zoom in figure is required to judge the result.
I was expecting to see three discs in each plate of treatment and this experiment shall be replicated three times, before analyzing the data and arriving into a conclusion. Please comment on your replication method.

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Pharmacology, toxicology, molecular endocrinology

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.

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23 Oct 2018 | for Version 1

Md Fakruddin, Laboratory of stem cell stress, International Research Center for Medical Sciences, Kumamoto University, Kumamoto , Japan

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Not Approved

In the title, it should be mention ethanol extract, as the authors only evaluate ethanol extract of the leaves.
In the figure 1, the quality is poor, so it seems the positive control do not have any inhibition zone!
Only one strain of each bacteria was tested, it would be more authentic if more types of strains as well as some wild or environmental strains were tested.
MIC/MBC of the extract against these bacteria should be determined.
Data of this study should be compared with similar data published already. It is understood, that there may be no data of this particular species, but other species of terminalia can be consulted.

Competing Interests

No competing interests were disclosed.

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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20 Sep 2018 | for Version 1

Edwin Setiawan, Department of Biology, Faculty of Natural Sciences, Sepuluh Nopember Institute of Technology, Surabaya, Indonesia

27 Views Cite this report Responses(0)

Approved

The study is designed appropriately and and technical procedure has been taken sufficiently to answer the research question. Furthermore, methods and analyses process also sufficient to be replicated by readers. In addition, statistical method that used is also appropriate. They author need to highlighted and emphasized on the results and discussion with expanding their discussion with comparing another result that similar to this study.

In the result and discussion part, it is better to compare phenolic and flavonoid content of Terminalia catappa to other plant extract that close to Terminalia catappa if possible. Or in other words, comparing other similar research that used plant bioactive compound for antibacterial to research. Therefore, this result and discussion part could be expanded comprehensively.

In addition, simple and short explanation on how the mechanism flavonoid and phenolic inhibit bacterial growth can be added in this part.

Competing Interests

No competing interests were disclosed.

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

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[1] 1. Najiah M, Aqilah NI, Lee KL, et al.: Massive mortality associated with Streptococcus agalactiae infection in cage-cultured red hybrid tilapia Oreochromis niloticus in Como River, Kenyir Lake, Malaysia. J Biol Sci. 2012; 12(8): 438–442. Publisher Full Text

[2] 2. Na-Phatthalung P, Chusri S, Suanyuk N, et al.: In vitro and in vivo assessments of Rhodomyrtus tomentosa leaf extract as an alternative anti-streptococcal agent in Nile tilapia (Oreochromis niloticus L.). J Med Microbiol. 2017; 66(4): 430–439. PubMed Abstract | Publisher Full Text

[3] 3. Saikia DJ, Chattopadhyay P, Banerjee G, et al.: Time and dose dependent effect of Pseudomonas aeruginosa infection on the scales of Channa punctata (Bloch) through light and electron microscopy. Turk J Fish Aquat Sci. 2017; 17(5): 871–876. Publisher Full Text

[4] 4. Baldissera MD, Souza CF, Santos RCV, et al.: Pseudomonas aeruginosa strain PA01 impairs enzymes of the phosphotransfer network in the gills of Rhamdia quelen. Vet Microbiol. 2017; 201: 121–125. PubMed Abstract | Publisher Full Text

[5] 5. Thomas J, Thanigaivel S, Vijayakumar S, et al.: Pathogenecity of Pseudomonas aeruginosa in Oreochromis mossambicus and treatment using lime oil nanoemulsion. Colloids Surf B Biointerfaces. 2014; 116: 372–377. PubMed Abstract | Publisher Full Text

[6] 6. Grema HA, Geidam YA, Suleiman A, et al.: Multi-Drug Resistant Bacteria Isolated from Fish and Fish Handlers in Maiduguri, Nigeria. Int J Anim Vet Adv. 2015; 7(3): 49–54. Reference Source

[7] 7. Samanidou VF, Evaggelopoulou EN: Analytical strategies to determine antibiotic residues in fish. J Sep Sci. 2007; 30(16): 2549–2569. PubMed Abstract | Publisher Full Text

[8] 8. Uchida K, Konishi Y, Harada K, et al.: Monitoring of Antibiotic Residues in Aquatic Products in Urban and Rural Areas of Vietnam. J Agric Food Chem. 2016; 64(31): 6133–8. PubMed Abstract | Publisher Full Text

[9] 9. He X, Deng M, Wang Q, et al.: Residues and health risk assessment of quinolones and sulfonamides in cultured fish from Pearl River Delta, China. Aquacult. 2016; 458: 38–46. Publisher Full Text

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Antimicrobial activity of Terminalia catappa brown leaf extracts against Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853

Abstract

Keywords

Introduction

Methods

Site and time

Bacterial strains and culture condition

Plant materials

Extraction procedure

Phytochemical content

Antibacterial activity assay

Data analysis

Results and discussion

Figure 1. Inhibition zone of Terminalia catappa brown leaves ethanolic extract against Staphylococcus aureus and Pseudomonas aeruginosa.

Table 1. Inhibitory zone diameter (in mm) of Staphylococcus aureus and Pseudomonas aeruginosa after treated with different concentration of brown leaves of T. catappa ethanolic extract.

Conclusion

Data availability

Grant information

Acknowledgments

References

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