Sahar M. A. ElSheikh1, Awad M. Abdel-Rahim2and Abasher A. Abasher2
1. University of Al sheikh Abdallah albdrie, Sudan
Email: sahargado97@gmail.com
2. Faculty of Sciences, University of Gezira, Sudan
HNSJ, 2022, 3(8); https://doi.org/10.53796/hnsj3825
Published at 01/08/2022 Accepted at 25/07/2022
Abstract
The Spearmint , Guava and Sabar are one of the plants found in the North African region. These plants have medicinal and pharmaceutical properties as well as antioxidants and food flavorings, and are used in skin care products and cosmetics. The aim of paper research is to study the antibacterial properties of Spearmint , Guava and Sabar leaves and stems. Against the following bacterial: E. coli and staphylococcus . auras .This study was conducted at the University of Gezira 2019/20 by selecting the bacterial number of colons .as well as the spore germination of the bacterial using the nutritional medium that nutritional medium that Nutrient Agar ( NA) , of the leaves and stems Photochemical screening of Guava, Spearmint and Sabar leaves. Spearmint , Guava and Sabar were brought from the local market of wad Madani, sample of bacteria obtained from the laboratory of the University of Gezira. Four concentration ( 25 ,50 , 75 , 100% ) were made from the aqueous extract of the leaves and stems of Spearmint and Guava . and only Sabar leaves. The result showed that the aqueous extracts of leaves and stems of Spearmint, Guava, and Sabar leaves showed the effected on E. coli the results showed that the higher concentration were highly effected. The Spearmint leaf extracts were found to decrease the number of colonies from 88 to 1and from 24 to 1 at the higher (75 and 100%), concentration respectively The effect of the different concentration of Guava and Sabar leaf extracts on E. coli are showing in Table (2, 3 ). The results indicated that the extracts are less effected compared to the Spearmint leaf extracts the Guava and the Sabar leaf extracts both gave 14 similar of control at the higher concentration (100%) respectively
However the Spearmint stem extracts are more effective allowing 11.7 colonies to grow compare to 12.9 colonies by stem extracts of Guava. the number of colonies of the bacteria S. auras are showing Table respectively the leaf extracts of Sabar were less effective allowing 12.7 colonies to grow, compared to that of Spearmint and Guava which allowed only 10.7 and 9.9 colonies to grow are showing the effects of the Guava leaf and stem extracts on the inhibition zone of E. coli both extracts were found highly effective. The inhibition zone was increased from 4.2 mm to 12.7 mm by the leaf extracts and from 7.2mm to 15.9 mm by the stem extracts .photochemical screening of the leaves the Spearmint , Guava and Sabar leaves were found to content all the compared except sterols .The Spearmint stems contain tannins , flavonids and alkaloid but not saponin are sterols. The Guava stem contains only Tannin and flavonoid but no saponin , alkaloid and sterol are detected
Plants have formed the basis for traditional medicine systems in most societies and have been used for thousands of years. It was estimated that there are 250000 – 500000 species of plants on earth (Odugbemi, 2008). However, today, the WHO has estimated that about 80% of the world’s inhabitants rely mainly on traditional medicines for their primary health care, where plant based systems still play a vital role in health care (Mullholand, 2000). Due to the wide diversity of botanical and large number of species, Sudanese medical plants can be considered as very promising candidates as bioactive agents (Abdel Daim, (2001; Abdel-Rahim, and Idris, 2010)
Spearmint is species of mint native to North Africa, Egypt and Morocco. It is an invasive species in Great Lakes region where it was first sighted in 1843. Spearmint has long tradition medicinal use. It was taken as a tea to treat general digestive problems. Spearmint is widely used in commercially manufactured product, cooking and medicine for its aromatic and flavorsome qualities. The objectives of this study were to determine the chemical composition of the spearmint, to determine the physicochemical properties Guava is a small tropical tree that grows up to 35feet tall; it is widely grown for its fruit in tropics. Itis a member of the Myrtaceae family, with about133 genera and more than 3,800 species. The leaves and bark of P. guajava tree have a long history of medicinal uses that are still employed today (Nwinyi et al., 2008). Sabar is in sub-Saharan Africa, the Arabian Peninsula and a number of Indian Ocean islands. They occupy many different kinds of natural habitat, from forest to exposed rock surfaces, but they are absent from the moist lowland forests of (Kamath et al.,2008)
In the view of the immense medicinal importance of P. guajava plant evidenced in the various studies mentioned above and also corroborated in a recent review article by Kamath et al. (2008). The medicinal plants display antioxidant and antimicrobial properties which can protect the human body against both cellular oxidation reactions and pathogens. Thus it is important to characterize different types of medicinal plants for their antioxidant and antimicrobial potentials (Mothana and Lindequist , 2005., Bajpai et al ,. 2005, Wojdylo et al 2007). Herbal medicine is the oldest from of medicine known to mankind. It was the mainstay of many early practiced form of medicine in the world today according to world health organization figures. While 25 to 50 % of the current pharmaceuticals are derived from plants, none are used as antimicrobials. Traditional healers have long used plants to prevent or cure infectious conditions plants are rich in a wide variety of secondary metabolites such as tannins, terpenoids, alkaloids, and flavonoids, which have been found in vitro to have antimicrobial properties. The structure and antimicrobial properties of the phytochemicals are also addressed . Since many of these compounds are currently available as unregulated botanical preparations and their use by the public is increasing rapidly, clinicians need to consider the consequences of patients self-medicating with these preparations
Materials and Methods
Preparation of plant part extracts:
The plants under test (Spearmint, Guava and Sabar) were collected from Wad Medani City, Sudan. The leaves and stems were washed in tap water, dried for 10 days at room temperature and blended into a powder using a mortar and pestle. Five concentrations (0.0, 25.0, 50.0, 75.0 and 100.0 mg/ml) were made by serial dilution of the different extracts with the medium in the flasks. All solutions were sterilized in an autoclave at 121C0 (15/b/in2) for 15 minutes and then cooled to room temperature.
Effect of the extracts on bacterial growth
Twenty-eight grams of an already prepared medium (Himedia Ltd.),were used per 1000ml of distilled water . the medium was dispensed into flasks (each containing 250ml ),and autoclaved at 1210 C (151 b/in2 ) for 15 minutes , then poured into sterile petri dishes, which were allowed to solidity and kept inverted into a refrigerator before use. The Cup –plate agar diffusion (Inhibition zone ) method
This method was used , using Nutrient Agar (NA). In this method 2ml of a standardized bacterial cell suspension (10×105) of E. coli or of Staphylococcus aurous were thoroughly mixed with 200ml of sterile molten nutrient agar, then the medium was distributed into sterile Petri-dishes and was left to solidity at room temperature for 24 hours. Sterile Whatsman glass fiber discs (No.5) were saturated with the extract then allowed to dry and transferred centrally on the surface of the solidified medium in each plate. The plates were then incubated at room temperature for 72hours and the inhibition zones were measured as described by Barry et al (1970).
Photochemical screening
photochemical screening to identify classes of secondary metabolites. Solvent extract of the plant material with help of different solvent (choroform, ethanol and Hcl). Photochemical analysis of the extract revealed presence of Saponin , Tannin , Flavonoid, Alkaloid, steroids and phenolic compound.
Experimental Result
The effect of the different concentration of the leaf plants extracts on both the number of colonies of E. coli were receded on Table( 1-3). Table (1) showed the effected on E. coli the results showed that the higher concentration were highly effected. The Spearmint leaf extracts were found to decrease the number of colonies from 88 to 1and from 24 to 1 at the higher 75 and 100%), concentration respectively (Table, 1).
Table (1): Effects of different concentrations of the Spearmint leaf on the number of colonies of E. coli
| concentration | Time (Hours) | |||
| 6 | 12 | 18 | 24 | |
| 0.0% | 62 | 80 | 88 | 90 |
| 25% | 50 | 68 | 76 | 80 |
| 50% | 30 | 60 | 62 | 54 |
| 75% | 7 | 10 | 12 | 18 |
| 100% | 1 | 1 | 1 | 1 |
The effect of the different concentration of Guava and Sabar leaf extracts on E. coli are showing in Table (2- 3 ). The results indicated that the extracts are less effected compared to the Spearmint leaf extracts the Guava and the Sabar leaf extracts both gave (4.5 and 2,7) from Guava leaf and (4.5 and 10.5) from sabar leaf at the higher concentration (75 -100%) respectively
Table (2):Effects of different concentrations of the Guava leaf on the number of colonies E. coli
| concentration | Time Hours | |||
| 6 | 12 | 18 | 24 | |
| 0.0% | 4.2 | 4.2 | 4.2 | 4.2 |
| 25% | 8.4 | 7.7 | 8.0 | 8.1 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 10.5 | 11.5 | 11.6 | 11.6 |
| 100% | 2 | 2.7 | 2.7 | 2.7 |
Table (3):Effects of different concentrations of the Sabar leaf on the number of colonies E. coli
| Concentration | Time (Hour) | |||
| 6 | 12 | 18 | 24 | |
| 0.0 | 4.5 | 4.5 | 4.5 | 4.5 |
| 25% | 7.4 | 7.7 | 8.0 | 8.01 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 10.5 | 11.5 | 11.4 | 11.4 |
| 100% | 10 | 10 | 10.3 | 10.5 |
The effect of the stem extracts of both Spearmint and Guava are shown in tables (4 and 5, respectively ). The results indicated that both extracts are effective. However the Spearmint extracts are more effective allowing 8.7 colonies to grow (table ,4) compare to 12.9 colonies by stem extracts of Guava ( table, 5).
Table (4): Effects of different concentrations of the Spearmint stem on the number of colonies of E. coli
|
|||||||||||||||||||||||||||||||||||
Table (5):Effects of different concentrations of the Guava stem on the number of colonies E. coli
| concentration | Time hours | |||
| 6 | 12 | 18 | 24 | |
| 0% | 7.2 | 7.2 | 7.2 | 7.2 |
| 25% | 8.4 | 7.7 | 8.0 | 8. 1 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 10.5 | 10.5 | 11.6 | 11.6 |
| 100% | 12 | 12.9 | 12.9 | 12.9 |
The effect of the leaf extracts of the three plant (spearmint , Guava and Sabar) on the number of colonies of the bacteria Staphy. auras are showing Table (6,7,and 8) respectively. The leaf extracts of the Spearmint ,Guava and Sabar found effective in reducing the number of colonies of Staphy. auras. However the leaf extracts of Sabar were less effective allowing 12.7 colonies to grow (Table,8), compared to that of Spearmint and Guava which allowed only 4.7 and 9.9 colonies to grow (Tables, 6 and7).
Fig (1) different of concentration to bacteria in plant extract steam
Table (6): Effects of different concentrations of the Spearmint leaf on the number of colonies Staphylococcus. aurous
| concentration | Time Hours | |||
| 6 | 12 | 18 | 24 | |
| 0% | 5.1 | 5.1 | 5.1 | 5.1 |
| 25% | 7.4 | 7.7 | 8.0 | 8.01 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 8.5 | 8.5 | 9.6 | 10.6 |
| 100% | 4 | 4.7 | 4.7 | 4.7 |
Table (7): Effects of different concentrations of the Guava leaf on the number of colonies of Staphylococcus. aurous
| concentration | Time (Hours) | |||
| 6 | 12 | 18 | 24 | |
| 0% | 4 | 4 | 4 | 4 |
| 25% | 6.4 | 6.7 | 7.0 | 7.01 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 10.5 | 11.5 | 11.6 | 11.6 |
| 100% | 9.9 | 9.9 | 9.9 | 9.9 |
Table (8) Effects of different concentrations of the Sabar leaf on the number of colonies Staphylococcus .aurous
| concentration | Time Hours | |||
| 6 | 12 | 18 | 24 | |
| 0% | 6 | 6.1 | 6.1 | 6.1 |
| 25% | 7.3 | 7.6 | 8.0 | 8.02 |
| 50% | 9.2 | 9.5 | 9.9 | 10.0 |
| 75% | 10.5 | 11.3 | 11.6 | 11.6 |
| 100% | 12 | 12.7 | 12.7 | 12.7 |
Fig (2) different of concentration to plant extract to leave plant number coloni
Tables (9 ,10) showed the effects of stem extracts of both Spearmint and Guava and number of colonies Staphy. arouse respectively both extracts were found effective given (10.7 and 13.9) respectively for concentration (100%)
Table (9): Effects of different concentrations of the Spearmint stem on the number of colonies of Staphylococcus. aurous
| concentration | Time Hours | |||
| 6 | 12 | 18 | 24 | |
| 0% | 6.1 | 6.1 | 5.1 | 5.1 |
| 25% | 8.4 | 7.7 | 8.0 | 8.01 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 8.5 | 8.5 | 9.6 | 10.6 |
| 100% | 11.2 | 12.7 | 10.7 | 10.7 |
Fig (3) inhibition zone in plant extract leave
Table (10): Effects of different concentrations of the Guava stem on the number of colonies Staphylococcus. aurous
| concentration | Time Hours | |||
| 6 | 12 | 18 | 24 | |
| 0% | 5.5 | 5.5 | 5.5 | 5.5 |
| 25% | 6.4 | 6.7 | 7.0 | 7.01 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 10.5 | 11.5 | 11.6 | 11.6 |
| 100% | 13.9 | 13.9 | 13.9 | 13.9 |
| The effect of the different concentration of Spearmint leaf and stem extract on the inhibition zone of E. coli are shown in table (11 and 12) the results indicated that both the leaf and the stem extracts are very effective increasing the inhibition from (5.5 to 6.5) and (6.7 to 10.7) at the 8th day of incubation.
Table (11): Effects of different concentrations of the Spearmint leaf on the inhibition zone of E. coli
|
|||||||||||||||||||||||||||||||||||||||||
Table (12): Effects of different concentrations of the Spearmint stem on the inhibition zone of E. coli
|
|||||||||||||||||||||||||||||||||||
Tables (13 and 14) are showing the effects of the Guava leaf and stem extracts on the inhibition zone of E. coli both extracts were found highly effective. The inhibition zone was increased from 4.2 mm to 12.7 mm by the leaf extracts and from 7.2mm to 15.9 mm by the stem extracts
Table (13): Effects of different concentrations of the Guava leaf on the inhibition zone of E. coli
| concentration | Days | |||
| 2 | 4 | 6 | 8 | |
| 0% | 4.2 | 4.2 | 4.2 | 4.2 |
| 25% | 8.4 | 7.7 | 8.0 | 8. 1 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 10.5 | 11.5 | 11.6 | 11.6 |
| 100% | 12 | 12.7 | 12.7 | 12.7 |
Table (14):Effects of different concentrations of the Guava stem on the inhibition zone of E. coli
| concentration | Days | |||
| 2 | 4 | 6 | 8 | |
| 0% | 7.2 | 7.2 | 7.2 | 7.2 |
| 25% | 8.1 | 8.7 | 9.0 | 8. 8 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 13.5 | 13.5 | 13.6 | 13.6 |
| 100% | 15 | 15.9 | 15.9 | 15.9 |
The results on Table (15) are showing the effects of the Sabar leaf extracts of the inhibition zone of E. coli the results indicated that the extracts are also effective increasing the inhibition zone from 4.5 mm to 12.0 at the 8th days of incubation
Table (15). Effects of different concentrations of the Sabar on the inhibition zone of E. coli
| concentration | Days | |||
| 2 | 4 | 6 | 8 | |
| 0% | 4.5 | 4.5 | 4.5 | 4.5 |
| 25% | 7.4 | 7.7 | 8.0 | 8.01 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 10.5 | 11.5 | 11.4 | 11.4 |
| 100% | 12 | 12 | 12 | 12 |
The effect of the leaf and stem extracts of spearmint on the inhibition zone of Staphy. arouse are shown in Table (16, 17) respectively. The result showed an effective results increasing the zone from (5.1 to 9.7) and (5.1 to 10.7) extract at 8th days of incubation
Table (16): Effects of different concentrations of the Spearmint leaf on the inhibition zone of Staphylococcus. aurous
| concentration | Days | |||
| 2 | 4 | 6 | 8 | |
| 0% | 5.1 | 5.1 | 5.1 | 5.1 |
| 25% | 7.4 | 7.7 | 8.0 | 8.01 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 8.5 | 8.5 | 8.6 | 10.6 |
| 100% | 9.2 | 9.7 | 9.7 | 9.7 |
Table (17): Effects of different concentrations of the Spearmint stem on the inhibition zone of Staphylococcus. arouse
| concentration | Days | |||
| 2 | 4 | 6 | 8 | |
| 0% | 6.1 | 6.1 | 5.1 | 5.1 |
| 25% | 8.4 | 7.7 | 8.0 | 8.01 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 8.5 | 8.5 | 9.6 | 10.6 |
| 100% | 11.2 | 12.7 | 10.7 | 10.7 |
Fig (4) different concentration of plant extract steam
Tables (18 ,19). Are showing the effect of both the leaf and stem extracts of Guava on the inhibition zone of the bacterium Staphy. arouse the extracts were less effective compared to that for Spearmint , although increased the zones from( 4.0 mm to 9.9) only and (5.5 to 14.9) at the 8th day of incubation
Table (18)Effects of different concentrations of the Guava leaf on the inhibition zone of Staphylococcus
| concentration | Days | |||
| 2 | 4 | 6 | 8 | |
| 0% | 4 | 4 | 4 | 4 |
| 25% | 6.4 | 6.7 | 7.0 | 7.01 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 10.5 | 11.5 | 11.6 | 11.6 |
| 100% | 9.9 | 9.9 | 9.9 | 9.9 |
Table (19): Effects of different concentrations of the Guava stem on the inhibition zone of Staphylococcus. aurous
| concentration | Days | |||
| 2 | 4 | 6 | 8 | |
| 0% | 5.5 | 5.5 | 5.5 | 5.5 |
| 25% | 6.4 | 6.7 | 7.0 | 7.01 |
| 50% | 9.4 | 9.8 | 10.0 | 10.0 |
| 75% | 10.5 | 11.5 | 11.6 | 11.6 |
| 100% | 14.9 | 14.9 | 14.9 | 14.9 |
The effect of the leaf extracts of Sabar are showing in Table (20). From the result can be seen that they extracts are highly effective , increasing the zone from 6.1 mm to 12.7 mm at the 8th day of incubation
Table (20): Effects of different concentrations of the Sabar leaf on the inhibition zone of Staphylococcus. aurous
| concentration | Days | |||
| 2 | 4 | 6 | 8 | |
| 0% | 6 | 6.1 | 6.1 | 6.1 |
| 25% | 7.3 | 7.6 | 8.0 | 8.02 |
| 50% | 9.2 | 9.5 | 9.9 | 10.0 |
| 75% | 10.5 | 11.3 | 11.6 | 11.6 |
| 100% | 12 | 12.7 | 12.7 | 12.7 |
The photochemical screening of the leaves and stem of the Spearmint and Guava is stem on Table (21). However the table also contend photochemical screening of the Sabar leaves the Spearmint , Guava and Sabar leaves were found to content all the compared except sterols .The Spearmint stems contain tannins , flavonids and alkaloid but not saponin are sterols. The Guava stem contains only Tannin and flavonoid but no saponin , alkaloid and sterol are detected
Table(21): photochemical screening of the leaves and stem of the spearmint
| Sabar leaves | Guava stem | Guava leaves | Spearmint stem | Spearmint leaves | Class chemical |
| ++ | – | + | + | ++ | Saponins |
| + | + | ++ | + | + | Tannins |
| + | + | +++ | + | ++ | Flavonoids |
| + | – | – | + | ++ | Alkaloids |
| – | – | – | – | – | Sterols |
+ indicated the presences of the class
_ indicated that, the class was not tested
Discussions
The use of plants and their extracts as remedies for curing many diseases have stimulated interest for investigating the presence of antimicrobial substances (Suliman et al., 2008 and Abdel-Rahim et al., 2010). However, the antimicrobial of many plant extracts have already well documented The effect of the leaf extracts of Spearmint , Guava and Sabar on number of colony and spore germination for tow bacteria E. coli and staphylococcus auras are tested in the present study. The results of the study indicated that all the extracts were effective. The leaves extracts of the Spearmint are very effective in reducing the number of coloni of both bacteria while ,the extracts of the leaf and the stem of Guava were less effective. The effects of the of the extracts on the spore germination of the bacteria E.coli indicated that the Spearmint leaf extracts are the best in reducing the number of coloni giving followed by Sabar extracts, while the guava extracts are least effective. Extracts of many plant species were reported to have antibacterial activities. In Sudan many studies were carried out for testing the antimicrobial activities of some medicinal plants. The extracts tested of 3 plants for antimicrobial activities
References:
Abdal-rahim,A.M., Al-jali, Z.I. and ALmismari , S.S.(1997) . factors affecting the growth and aflatoxin production by the two fungi A.flavus and A.parasiticus 6th Arab congress on plant protection, Beirut, Lebanon OCT.1997
Abdel-RahimA.M.,Mohammed-ali, R.J. and Al-jali, Z.I. (2002).effect of natural products on fungal growth and aflatoxin production in A.flavus and A.parasiticus. the 3rd international conference of fungi , cairo Egypt OCT. 2002.
Abdel -Rahim, A.M. (2005). Aflatoxin. Gezira publishing company ltd wad medani sudan
Abdal-Rahim, A.M and Idris, F.A. (2010). Survival of staphylococcus aureus and E.coli on cotton fabrics treated with extract of garad(Acacia nilotical). Gezira j.ofeng . & Applied sci. 5 (2) : 127 -134.
Abdal-Rahim, A.M. Alsheikh,M.S, and Suleiman, A.E.(2010). Aflatoxin contamination in some cereal and legume seed and their products in the Gezira state.5(2) 92-110
Abdel-Daim, Z.J.(2001).Z.J. phytochemical and microbial studies on some Senna species M.sc. Faculty of science, University of Khartoum .
Ahemd S.S.A. (2004). Phytochemical . antimalarial, Mollucicigdal and Antimicrobial activity of selected Sudanese Medicinal plants with Emphasis on Nigella sativa L.seeds. ph.D. thesis, University of Gezira .
AOAC,(2010) Association of Official Analysis Chemists
Bajpai, M, Pande, A, Tewari,sk and parkask, D,(2005). Phenolic content and antioxidant activity of some food and medicinal plants. International journal of food sciences and Nutrition 56(4): 287-291.
Kamath Jv.Rahul (2008). Psiddium guajava . a review int j. green pharm
Mothana, R.AA. and Lindequist, U,(2005). Antimicrobial activity of some medicinal plants of the Island Sogotra, journal of Ethno pharmacology,96:177- 181
Mullholand, v. (2000) application of molecular biology methods to the detection and characterization of organisms of agricultural.
Nwinyi Oc, Chined. U. (2008). Evaluation of antibacterial activity of psidum guajava and gongronema latifolium. J. med plants Res. 2(8) : 189- 192
Odugbemi, T.( 2008). A textbook of Medicinal plants in Nigeria. Tolu PRESS Lagos p.23-97
Sulieman, A.E, Ahmed, H.E. and Abdel Rahim, A.M. (2008). The chemical composition of fenugreek ( trigonella foenum graceum L) and the antimicrobial properties of its seed oil. Thesis University of Gezira
Wojdylo A, Oszmianski j, and Czemerys R. (2007). Antioxidant activity and phenolic compounds in 32 selected herbs . food chem. 105, 940-949.
Arabic 
English