Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of <i>Combretum molle</i> (<i>Combretaceae</i>)

Yakai Fawai; Dawe Amadou; Wangso Albert; Fanta Angele; Moussa Djaouda; Taira Vincent; Doudja Chantal; Loura Benoit

doi:doi:10.11648/j.ajcbe.20230701.12

| Peer-Reviewed

Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae)

Yakai Fawai, Dawe Amadou, Wangso Albert, Fanta Angele, Moussa Djaouda, Taira Vincent, Doudja Chantal, Loura Benoit

Published in American Journal of Chemical and Biochemical Engineering (Volume 7, Issue 1)

Received: 14 May 2023 Accepted: 8 June 2023 Published: 21 June 2023

Views: Downloads:

Download PDF

Share This Article

Twitter
Linked In
Facebook

Abstract

Combretum molle species are extensively used in traditional medicine in the Northern regions of Cameroon against inflammation, infections, diabetes, malaria, bleeding, diarrhea, digestive disorders and others as a diuretic as back pain. This study was conducted to evaluate the antibacterial effects of crude methanolic extract, ethyl acetate extract and isolated compounds from the fruits of Combretum molle using the Muller-Hinton solid medium disc method. Isolation of secondary metabolites from the fruits of Combretum molle was done by the means of usual chromatographic techniques and their structures were confirmed on the basis of spectroscopic data and those available in literature. The first phytochemical exploration of the fruits of Combretum molle afforded sitosterol-3-O-β-D-glucoside (1), oleanolic acid (2), mollic acid (3), mollic acid-3-O-β-D-glucoside (4), 3, 5-dihydroxy-4’,7-dimethoxyflavone (5), and 5,4’-dihydroxy-7-methoxyflavone (6). To the best of our knowledge, compounds (5) and (6) are isolated and reported for the first time from this plant. At the concentration of 25mg/ml methanolic extract, ethyl acetate extract and compounds 3, 4 and 5 presented good antibacterial potential on Salmonella typhimurium (ATCC13311) strain with diameters of the inhibition zone of 18.2±0.1mm, 15.3±0.2mm, 14.2± 0.3mm, 20.2±0.1mm and 15.1± 0.2mm respectively. Also compound 4 and 6 were sensitive on Pseudomonas aeruginosa (ATCC27853) with diameters of inhibition of 18.2±0.2 mm and 14.1± 0.4mm respectively. The results were compared to TE30 (tetracyclin), CIP5 (ciprofloxacin), C30 (chloremphinincol) and NA30 (nalidicic acid) used as references. These results are in agreement with the enthnobotanical uses of the plant and some previous reports on pharmacological activities such as anti-inflammatory, antioxidant, anticancer effects of some isolates and their related previously isolated and identified from several plants.

Published in	American Journal of Chemical and Biochemical Engineering (Volume 7, Issue 1)
DOI	10.11648/j.ajcbe.20230701.12
Page(s)	7-14
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2023. Published by Science Publishing Group

Keywords

Combretum molle, Fruit, Triterpenes, Flavonoid, Antibacterial Effect

References

[1]	De Leo. M, De Tommasi. N, Sanogo. R, D’Angelo. V, Germano. M. P, Bisignano. G, Braca. A, “Triterpenoid saponins from Pteleopsis suberosa stem bark”. Phytochemistry, 2006, 60, 2623-2629.
[2]	Rademan. S, Lall. N, “Combretum molle, in Underexplored Medicinal Plants from Sub-Saharan Africa: Plants with Therapeutic Potential for Human Health”, Elsevier, 2020; pp. 67-76.
[3]	Keay. R, W, Y., “Trees of Nigeria. 3rd eds”. Clavevdo Press Oxford, 1989; Pp 146-216.
[4]	Kerharo. J, Adams. J. G, “Pharmacopée sénégalaise traditionnelle: plantes médicinales et toxiques”, Ed Vigot et Frères, Paris, 1974, 1011.
[5]	Reham. Z. H, Shaden. E. Al-Motaani, Tarek. Al-Talhi, “Therapeutic and Ameliorative Effects of Active Compounds of Combretum molle in the Treatment and Relief from Wounds in a Diabetes Mellitus Experimental Model, Coatings, 2021; 11 (3), 324.
[6]	Yéo. D, Koffi. E, Bidié. A. P, Tako. N. A, Bahi. C, Méité. S, Djaman. A. J, Guéde-Guina. F, “In Vitro Anticholinesterase and Inhibitory Effects of the Aqueous Extract of Combretum molle (Combretaceae) Leaf on Rabbit Breathing”, Tropical Journal of Pharmaceutical, 2010; 9 (5) pp 469-473.
[7]	Fyhrquist. P, Mwasumbi. L, Vuorela. P, Vuorela. H, Hiltunen. R, Murphy. C, “Preliminary antiproliferative effects of some species of Terminalia, Combretum and Pteleopsis collected in Tanzania on some human cancer cell lines”, Fitoterapia, 2006; 77: 358–366.
[8]	Parusnath. M, Naidoo. Y, Singh. M, Rihan. H, Dewir. Y. H, “Phytochemical Composition of Combretum molle (R. Br. ex G. Don.) Engl. & Diels Leaf and Stem Extracts”, Plants, 2023; 12, 1702.
[9]	Mirriam. E, Nyenje, Roland. N. N, “Bioactivity of the acetone extract of the stem bark of Combretum molle on selected bacterial pathogens: Preliminary phytochemical screening”, Journal of Medicinal Plants Research, 2012; Vol. 6 (8), pp. 1476-1481.
[10]	Bolou. G. E. K, Attioua. B, N’guessam. A, C, Coulibaly. A, N’Guessan. J. D, Djaman. A. J, “In vitro evaluation of antibacterial activity of Terminalia glaucescens planch extract on Salmonella typhi and Salmonella typhimurium”, Newsletter of Liege Sciences Royal Society, 2011; vol. 80, P. 772-790.
[11]	Bauer. A. W, Kirby. W. M. M, Sherris. T. C, Truck. M, “Antibiotical susceptibility testing by a standardized single disc method”. American journal of clinical pathology, 1966; Vol. 45, P. 493-496.
[12]	Barry. A. L, Thornsberry. C, “Susceptibility test, diffusion test procedure”, American journal of clinical pathology, 1985; Vol. 19, P. 492-500.
[13]	FSM (French Society of Microbiology), Recommendations of Antibiogram committee of French Society of Microbiology, 2008; P. 49.
[14]	Adesokan. A. A, Akanji. M. A, “Antibacterial potentials of aqueous extract of Enantia chlorantha stem barck”, African journal of Biotechnology, 2007; P. 2502-2505.
[15]	Doughari. J. H, Pukuma. M. S, De. N, “Antibacterial effects of blanites aegyptiaca L. Drel and Moringa oleifera Lam. On Salmonella typhi”, African journal of biotechnology, 2007; Vol. 6 (19), P. 2212-2215.
[16]	Swadesh. S, Amrita. P, Anirban. C, Santanu. P, “A novel compound β-sitosterol-3-O-β-D-glucoside isolated from Azadirachta indica effectively induces apoptosis in leukemic cells by targeting G0/G1 populations”, Indian Journal of Biochemistry & Biophysics, Vol. 57, 2020; pp. 27-32.
[17]	Mofiz. U. K. N. M, Sagar. H, Md, “Scopoletin and β-sitosterol glucoside from roots of Ipomoea digitata “, Journal of Pharmacognosy and Phytochemistry, 2015; 4 (2): 05-07.
[18]	Castellano. J. M, Ramos-Romero. S, Perona. J. S, “Oleanolic Acid: Extraction, Characterization and Biological Activity”, Nutrients 2022; 14, 623.
[19]	Isaac. S. G, Corinne. C, Richard. T. S, Emmanuel. T, Alex. De T. A, Paule. S, Brigitte. V, Sophie. L, Celine. H, Armel. H. N. K, Jean-Marc. M, “A new phenyl alkyl ester and a new combretin triterpene derivative from Combretum fragrans F. Hoﬀm (Combretaceae) and antiproliferative activity”, Open Chemistry, 2020; 18: 1523–1531.
[20]	Manish. K. G, Sanjana. S, Adarsh. K. C, Kishore. B, Sosmitha. G, Ajaikumar. B. K, Vikash. K. D, Latha. R, “Antioxidant, anti-tyrosinase and anti-inflammatory activities of 3,5-dihydroxy-4′,7-dimethoxyflavone isolated from the leaves of Alpinia nigra”. Phytomedicine Plus, 2021; 1:100097.
[21]	Kerkatou. M, Redouane-Salah. A, León. F, Brouard. I, Mosset. P, Menad. A, Ameddah. S, Benayache. S, Bermejo. J, Benayache. F, “Secondary Metabolites and Antioxidant Activity of Limonium duriusculum (de Girard) Kuntze Extracts”, Asian J. Chem, 2016; 28 (12), 2695-2700.
[22]	CA-FSM/EUCAST (Antibiogram committee of French Society of Microbiology). European Committee on Antimicrobial Susceptibility Testing (CA-SFM/EUCAST), 2016: 117.
[23]	Dabole. B, Matcheme. M, Moussa. D, Abdou. T, W, Bouba. R, Meli. A, Loura. B, “Antibacterial Effects of Chemical Constituents Isolated from the Roots of Cordia myxa (Boraginaceae) on Salmonella typhi (Enterobacteriaceae)”, American Journal of Chemical and Biochemical Engineering. Vol. 5, No. 1, 2021, pp. 1-7.
[24]	Xiao-Nian. X, Fang. W, Yi-Ting. Y, Jing. L, Yue-Zhen. L, Xing. Y, “Antibacterial activity and Mode of action of Dihydromyricetin from Ampelopsis”. Molecules, 2019; 24 (15): 2831.

Cite This Article

Plain Text BibTeX RIS

APA Style

Yakai Fawai, Dawe Amadou, Wangso Albert, Fanta Angele, Moussa Djaouda, et al. (2023). Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae). American Journal of Chemical and Biochemical Engineering, 7(1), 7-14. https://doi.org/10.11648/j.ajcbe.20230701.12

Copy | Download

ACS Style

Yakai Fawai; Dawe Amadou; Wangso Albert; Fanta Angele; Moussa Djaouda, et al. Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae). Am. J. Chem. Biochem. Eng. 2023, 7(1), 7-14. doi: 10.11648/j.ajcbe.20230701.12

Copy | Download

AMA Style

Yakai Fawai, Dawe Amadou, Wangso Albert, Fanta Angele, Moussa Djaouda, et al. Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae). Am J Chem Biochem Eng. 2023;7(1):7-14. doi: 10.11648/j.ajcbe.20230701.12

Copy | Download

@article{10.11648/j.ajcbe.20230701.12,
  author = {Yakai Fawai and Dawe Amadou and Wangso Albert and Fanta Angele and Moussa Djaouda and Taira Vincent and Doudja Chantal and Loura Benoit},
  title = {Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae)},
  journal = {American Journal of Chemical and Biochemical Engineering},
  volume = {7},
  number = {1},
  pages = {7-14},
  doi = {10.11648/j.ajcbe.20230701.12},
  url = {https://doi.org/10.11648/j.ajcbe.20230701.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajcbe.20230701.12},
  abstract = {Combretum molle species are extensively used in traditional medicine in the Northern regions of Cameroon against inflammation, infections, diabetes, malaria, bleeding, diarrhea, digestive disorders and others as a diuretic as back pain. This study was conducted to evaluate the antibacterial effects of crude methanolic extract, ethyl acetate extract and isolated compounds from the fruits of Combretum molle using the Muller-Hinton solid medium disc method. Isolation of secondary metabolites from the fruits of Combretum molle was done by the means of usual chromatographic techniques and their structures were confirmed on the basis of spectroscopic data and those available in literature. The first phytochemical exploration of the fruits of Combretum molle afforded sitosterol-3-O-β-D-glucoside (1), oleanolic acid (2), mollic acid (3), mollic acid-3-O-β-D-glucoside (4), 3, 5-dihydroxy-4’,7-dimethoxyflavone (5), and 5,4’-dihydroxy-7-methoxyflavone (6). To the best of our knowledge, compounds (5) and (6) are isolated and reported for the first time from this plant. At the concentration of 25mg/ml methanolic extract, ethyl acetate extract and compounds 3, 4 and 5 presented good antibacterial potential on Salmonella typhimurium (ATCC13311) strain with diameters of the inhibition zone of 18.2±0.1mm, 15.3±0.2mm, 14.2± 0.3mm, 20.2±0.1mm and 15.1± 0.2mm respectively. Also compound 4 and 6 were sensitive on Pseudomonas aeruginosa (ATCC27853) with diameters of inhibition of 18.2±0.2 mm and 14.1± 0.4mm respectively. The results were compared to TE30 (tetracyclin), CIP5 (ciprofloxacin), C30 (chloremphinincol) and NA30 (nalidicic acid) used as references. These results are in agreement with the enthnobotanical uses of the plant and some previous reports on pharmacological activities such as anti-inflammatory, antioxidant, anticancer effects of some isolates and their related previously isolated and identified from several plants.},
 year = {2023}
}

Copy | Download

TY  - JOUR
T1  - Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae)
AU  - Yakai Fawai
AU  - Dawe Amadou
AU  - Wangso Albert
AU  - Fanta Angele
AU  - Moussa Djaouda
AU  - Taira Vincent
AU  - Doudja Chantal
AU  - Loura Benoit
Y1  - 2023/06/21
PY  - 2023
N1  - https://doi.org/10.11648/j.ajcbe.20230701.12
DO  - 10.11648/j.ajcbe.20230701.12
T2  - American Journal of Chemical and Biochemical Engineering
JF  - American Journal of Chemical and Biochemical Engineering
JO  - American Journal of Chemical and Biochemical Engineering
SP  - 7
EP  - 14
PB  - Science Publishing Group
SN  - 2639-9989
UR  - https://doi.org/10.11648/j.ajcbe.20230701.12
AB  - Combretum molle species are extensively used in traditional medicine in the Northern regions of Cameroon against inflammation, infections, diabetes, malaria, bleeding, diarrhea, digestive disorders and others as a diuretic as back pain. This study was conducted to evaluate the antibacterial effects of crude methanolic extract, ethyl acetate extract and isolated compounds from the fruits of Combretum molle using the Muller-Hinton solid medium disc method. Isolation of secondary metabolites from the fruits of Combretum molle was done by the means of usual chromatographic techniques and their structures were confirmed on the basis of spectroscopic data and those available in literature. The first phytochemical exploration of the fruits of Combretum molle afforded sitosterol-3-O-β-D-glucoside (1), oleanolic acid (2), mollic acid (3), mollic acid-3-O-β-D-glucoside (4), 3, 5-dihydroxy-4’,7-dimethoxyflavone (5), and 5,4’-dihydroxy-7-methoxyflavone (6). To the best of our knowledge, compounds (5) and (6) are isolated and reported for the first time from this plant. At the concentration of 25mg/ml methanolic extract, ethyl acetate extract and compounds 3, 4 and 5 presented good antibacterial potential on Salmonella typhimurium (ATCC13311) strain with diameters of the inhibition zone of 18.2±0.1mm, 15.3±0.2mm, 14.2± 0.3mm, 20.2±0.1mm and 15.1± 0.2mm respectively. Also compound 4 and 6 were sensitive on Pseudomonas aeruginosa (ATCC27853) with diameters of inhibition of 18.2±0.2 mm and 14.1± 0.4mm respectively. The results were compared to TE30 (tetracyclin), CIP5 (ciprofloxacin), C30 (chloremphinincol) and NA30 (nalidicic acid) used as references. These results are in agreement with the enthnobotanical uses of the plant and some previous reports on pharmacological activities such as anti-inflammatory, antioxidant, anticancer effects of some isolates and their related previously isolated and identified from several plants.
VL  - 7
IS  - 1
ER  -

Copy | Download

Author Information

Yakai Fawai

Department of Chemistry, Higher Teachers Training College, University of Maroua, Maroua, Cameroon
Dawe Amadou

Department of Chemistry, Higher Teachers Training College, University of Maroua, Maroua, Cameroon
Wangso Albert

Department of Chemistry, Higher Teachers Training College, University of Maroua, Maroua, Cameroon
Fanta Angele

Department of Chemistry, Higher Teachers Training College, University of Maroua, Maroua, Cameroon
Moussa Djaouda

Department of Biological Sciences, Faculty of Science, University of Maroua, Maroua, Cameroon
Taira Vincent

Department of Chemistry, Higher Teachers Training College, University of Maroua, Maroua, Cameroon
Doudja Chantal

Department of Chemistry, Higher Teachers Training College, University of Maroua, Maroua, Cameroon
Loura Benoit

Department of Chemistry, Faculty of Science, University of Maroua, Maroua, Cameroon

Download PDF

Sections

Plain Text BibTeX RIS

APA Style

Yakai Fawai, Dawe Amadou, Wangso Albert, Fanta Angele, Moussa Djaouda, et al. (2023). Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae). American Journal of Chemical and Biochemical Engineering, 7(1), 7-14. https://doi.org/10.11648/j.ajcbe.20230701.12

Copy | Download

ACS Style

Yakai Fawai; Dawe Amadou; Wangso Albert; Fanta Angele; Moussa Djaouda, et al. Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae). Am. J. Chem. Biochem. Eng. 2023, 7(1), 7-14. doi: 10.11648/j.ajcbe.20230701.12

Copy | Download

AMA Style

Yakai Fawai, Dawe Amadou, Wangso Albert, Fanta Angele, Moussa Djaouda, et al. Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae). Am J Chem Biochem Eng. 2023;7(1):7-14. doi: 10.11648/j.ajcbe.20230701.12

Copy | Download

@article{10.11648/j.ajcbe.20230701.12,
  author = {Yakai Fawai and Dawe Amadou and Wangso Albert and Fanta Angele and Moussa Djaouda and Taira Vincent and Doudja Chantal and Loura Benoit},
  title = {Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae)},
  journal = {American Journal of Chemical and Biochemical Engineering},
  volume = {7},
  number = {1},
  pages = {7-14},
  doi = {10.11648/j.ajcbe.20230701.12},
  url = {https://doi.org/10.11648/j.ajcbe.20230701.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajcbe.20230701.12},
  abstract = {Combretum molle species are extensively used in traditional medicine in the Northern regions of Cameroon against inflammation, infections, diabetes, malaria, bleeding, diarrhea, digestive disorders and others as a diuretic as back pain. This study was conducted to evaluate the antibacterial effects of crude methanolic extract, ethyl acetate extract and isolated compounds from the fruits of Combretum molle using the Muller-Hinton solid medium disc method. Isolation of secondary metabolites from the fruits of Combretum molle was done by the means of usual chromatographic techniques and their structures were confirmed on the basis of spectroscopic data and those available in literature. The first phytochemical exploration of the fruits of Combretum molle afforded sitosterol-3-O-β-D-glucoside (1), oleanolic acid (2), mollic acid (3), mollic acid-3-O-β-D-glucoside (4), 3, 5-dihydroxy-4’,7-dimethoxyflavone (5), and 5,4’-dihydroxy-7-methoxyflavone (6). To the best of our knowledge, compounds (5) and (6) are isolated and reported for the first time from this plant. At the concentration of 25mg/ml methanolic extract, ethyl acetate extract and compounds 3, 4 and 5 presented good antibacterial potential on Salmonella typhimurium (ATCC13311) strain with diameters of the inhibition zone of 18.2±0.1mm, 15.3±0.2mm, 14.2± 0.3mm, 20.2±0.1mm and 15.1± 0.2mm respectively. Also compound 4 and 6 were sensitive on Pseudomonas aeruginosa (ATCC27853) with diameters of inhibition of 18.2±0.2 mm and 14.1± 0.4mm respectively. The results were compared to TE30 (tetracyclin), CIP5 (ciprofloxacin), C30 (chloremphinincol) and NA30 (nalidicic acid) used as references. These results are in agreement with the enthnobotanical uses of the plant and some previous reports on pharmacological activities such as anti-inflammatory, antioxidant, anticancer effects of some isolates and their related previously isolated and identified from several plants.},
 year = {2023}
}

Copy | Download

TY  - JOUR
T1  - Antibacterial Effects of Crude Methanolic Extract, Ethyl Acetate Extract and Isolated Compounds from the Fruits of Combretum molle (Combretaceae)
AU  - Yakai Fawai
AU  - Dawe Amadou
AU  - Wangso Albert
AU  - Fanta Angele
AU  - Moussa Djaouda
AU  - Taira Vincent
AU  - Doudja Chantal
AU  - Loura Benoit
Y1  - 2023/06/21
PY  - 2023
N1  - https://doi.org/10.11648/j.ajcbe.20230701.12
DO  - 10.11648/j.ajcbe.20230701.12
T2  - American Journal of Chemical and Biochemical Engineering
JF  - American Journal of Chemical and Biochemical Engineering
JO  - American Journal of Chemical and Biochemical Engineering
SP  - 7
EP  - 14
PB  - Science Publishing Group
SN  - 2639-9989
UR  - https://doi.org/10.11648/j.ajcbe.20230701.12
AB  - Combretum molle species are extensively used in traditional medicine in the Northern regions of Cameroon against inflammation, infections, diabetes, malaria, bleeding, diarrhea, digestive disorders and others as a diuretic as back pain. This study was conducted to evaluate the antibacterial effects of crude methanolic extract, ethyl acetate extract and isolated compounds from the fruits of Combretum molle using the Muller-Hinton solid medium disc method. Isolation of secondary metabolites from the fruits of Combretum molle was done by the means of usual chromatographic techniques and their structures were confirmed on the basis of spectroscopic data and those available in literature. The first phytochemical exploration of the fruits of Combretum molle afforded sitosterol-3-O-β-D-glucoside (1), oleanolic acid (2), mollic acid (3), mollic acid-3-O-β-D-glucoside (4), 3, 5-dihydroxy-4’,7-dimethoxyflavone (5), and 5,4’-dihydroxy-7-methoxyflavone (6). To the best of our knowledge, compounds (5) and (6) are isolated and reported for the first time from this plant. At the concentration of 25mg/ml methanolic extract, ethyl acetate extract and compounds 3, 4 and 5 presented good antibacterial potential on Salmonella typhimurium (ATCC13311) strain with diameters of the inhibition zone of 18.2±0.1mm, 15.3±0.2mm, 14.2± 0.3mm, 20.2±0.1mm and 15.1± 0.2mm respectively. Also compound 4 and 6 were sensitive on Pseudomonas aeruginosa (ATCC27853) with diameters of inhibition of 18.2±0.2 mm and 14.1± 0.4mm respectively. The results were compared to TE30 (tetracyclin), CIP5 (ciprofloxacin), C30 (chloremphinincol) and NA30 (nalidicic acid) used as references. These results are in agreement with the enthnobotanical uses of the plant and some previous reports on pharmacological activities such as anti-inflammatory, antioxidant, anticancer effects of some isolates and their related previously isolated and identified from several plants.
VL  - 7
IS  - 1
ER  -

Copy | Download