







































10.11648.j.ijhnm.20220802.11


    ISSN : 2693 6356 

2022 | Vol 5 | Issue 3 

 
 

Experimental Investigation of the Antimicrobial Effects of Terminalia avicennioides 

Extracts on Staphylococcus aureus Strains Resistant to Multiple Drugs 
Dr. Manjusha Vithhal Ahire 

Assistant Professor 
Homoeopathic Medical College & Hospital, Jalgaon 

 

Abstract: As the prevalence of antibiotic resistance rises, fewer effective treatments are available for infections caused by 

MRSA and other multidrug-resistant bacteria. As a result, there was a surge in interest in medicinal plant extracts as a potential 

source of novel phytochemicals for the treatment of infectious disorders. The purpose of this research was to identify the in 

vitro antibacterial activity of Terminalia avicennioides extracts against MRSA strains that have developed resistance to many 

drugs used in wound infections. Patients from Nigeria's Barau Dikko Teaching Hospital in Kaduna provided the wound swab 

samples. We used conventional phenotypic and genotypic identification techniques to isolate and characterize Staphylococcus 

aureus. Following established protocols, we determined the antimicrobial susceptibility profile of the Staphylococcus aureus 

isolates. Standard protocols were also followed to synthesize Terminalia avicennioides extracts and test them for antibacterial 

activity against MRSA in vitro. Staphylococcus aureus isolates exhibited resistance to a wide range of conventional antibiotics, 

from 8.18% to 100%, according to the susceptibility profile. Nevertheless, imipenem was effective against all of the isolates. 

Phytochemical studies conducted on the extracts, both qualitative and quantitative, have shown that they include tannin, 

alkaloids, flavonoids, cardiac glycoside, phenols, saponins, and terpenoids, but no anthraquinones. Terminalia avicennioides 

extracts shown a strong antimicrobial effect against MRSA isolates, with growth inhibition zones ranging from 16.28±10.45 - 

23.81±6.69 mm and a p-value less than 0.05. The range of the extracts' minimum inhibitory concentrations (MIC) was 56.2500 

± 29.1241 - 31.2500 + 22.16013 gm/ml, and there was no significant difference (p > 0.05). There was no significant difference 

(p > 0.05) in the minimum bactericidal concentration (MBC) of the extracts, which varied from 175,000 ± 64.2910 to 68.7500 

± 45.8063 mg/ml. Surprisingly, the antimicrobial properties of Terminalia avicennioides extracts show stronger inhibitory 

effects against MRSA strains, suggesting they might be developed and studied further for the treatment of wound infections.  

Keywords: Staphylococcus aureus, Multidrug Resistance, Wound, Antibacterial Terminalia avicennioides 

 

 

1. Introduction 
Staphylococcus aureus persists in evading antibiotic control 

efforts and has a history of developing resistance to new 

medications. There has been a worldwide pandemic of 

infections caused by Staphylococcus aureus strains that are 

resistant to antibiotics, and the rates of antimicrobial 

resistance are on the rise. are reducing the number of 

available treatments [1]. The global economic and health 

burden of multidrug-resistant (MDR) diseases is immense 

and terrible. It has only lately come to light that 

antimicrobial-resistant illnesses cause the deaths of around 

700,000 people every year [2-4]. Unrecognized costs of 

multidrug-resistant infections (MDR) are disproportionately 

high in underdeveloped countries like Nigeria. therapy of 

antibiotic-resistant diseases and related fatalities. 

Antimicrobial resistance infections are a growing problem 

in modern medicine, and many important variables, 

including shifting demography, increased international 

trade, and extreme weather events, are exacerbating the 

problem [3, 4]. It is well acknowledged that the majority of 

antibacterial agents on the market, particularly synthetic 

ones, have been improperly utilized and no longer work [5-

7]. So, the World Health Organization (WHO) said that we 

should be looking for new antibiotics that work against 

bacteria that are resistant to multiple drugs, as well as ones 

that don't cross- or co-resist with other antibiotic classes [3]. 

The world over, people have been using traditional herbal 

remedies to cure a variety of infectious ailments for 

thousands of years [8, 9], and it's interesting to note that 

medicinal plants are also seen as possible sources of novel  

 

antimicrobial compounds. Because chemically produced 

drugs often cause side effects and microbial resistance, 

ethnopharmaconosy has become the preferred method of 

drug discovery involving natural products, such as plant 

extracts (either as pure compounds or as standardized 

extracts) [10, 11]. The simplicity, low cost, and high clinical 

performance of traditional healing agents make them ideal 

for use in wound care. These treatments provide a more 

economical option for treating various wounds that are 

difficult to heal, such as burns, ulcers, and infected wounds. 

They have a variety of therapeutic actions that speed up the 

healing process and enhance the quality of the new skin 

[12]. The purpose of this research was to determine whether 

or not Terminalia avicennioides extracts had any 

antibacterial effect on MRSA, a kind of bacteria often seen 

in wounds, when cultured in a laboratory setting.  

 

2. Materials and Methods 

2.1. Ethical Consideration 

2.2. At the Barau Dikko Teaching Hospital, Kaduna State 

University, Kaduna, Nigeria, we got approval from the 

research ethics committee (Reference number: HREC: 

20-0004) to take swabs from patients' wounds in order to 

isolate Staphylococcus aureus. Kaduna State University, 

Barau Dikko Teaching Hospital, Nigeria. Patients 

diagnosed with wound infections were asked to fill out 

an informed consent form before any pertinent data or 

wound swab samples were collected. The nurses of the 



chosen hospital wards and units were informed of the 

study's purpose and the ethical committee's clearance 

before wound swabs were taken from patients. In order 

to educate the patients, a concise description of the 

research's goals and purposes was given. Patients were 

also made aware that they might choose to participate or 

not. Parents or guardians of children who have a wound 

infection are kindly asked to provide their consent on 

behalf of the children.Collection of Wound Swabs and 

Isolation of Staphylococcus aureus from the Wound 

Swabs A total of sixty wound swabs samples were 

collected from 

2.3. in and out patients with wound at Barau Dikko Teaching 

Hospital Kaduna, Nigeria Exudate or purulent or pus 

discharge were aseptically swabbed with sterile swab 

cotton tip and the cotton tip broke immediately into a 

sterile Brain Heart Infusion (BHI) broth in a universal 

bottle. The collection of the samples from the patients 

were carried out with the help of the hospital Nurses. The 

samples collected were then transported in ice packed 

thermo flasks to Kaduna State University Postgraduate 

Medical Microbiology Laboratory for isolation of 

Staphylococcus aureus isolates. 

All media were prepared according to manufacturer's 

instructions. All clinical samples collected were cultured 

aerobically for isolation of Staphylococcus aureus in the 

laboratory as described by Vallis et al. [13] and 

Cheesbrough [14]. The swab samples were first cultured 

aerobically in an enrichment medium (Brain Heart 

Infusion (BHI) broth) at 37°C for 24 hours. The broth 

cultures from the BHI broth were then Manitol Salt agar 

(MSA) plates for selective isolation of Staphylococcus 

aureus. Pure culture colonies of presumptive 

Staphylococcus aureus on MSA plates were further 

subculture aerobically on Baired Parker agar plates at 

37°C for 24 hours for morphological characteristics study 

of the isolates. Pure single colonies from this medium 

were subculture on nutrient agar slant and kept at 4°C for 

biochemical morphological and biochemical 

characterisation. 

2.4. Morphological and Biochemical 

Characterisation of Presumptive Staphylococcus 

aureus Isolates 

Biochemical characterisation of the pure isolates 

obtained was carried out as described by Aneja, Ochai 

and Kolhatkar, and Cheesbrough [14-16]. Motility, 

catalase, coagulase, hemolysis, citrate utilization, methyl 

red, Voges-Proskauer, indole, and sugars (lactose, 

mannitol and sucrose) fermentation test were carried out 

for identification of Staphylococcus aureus isolates. 

2.5. Molecular Identification of Staphylococcus aureus 

2.5.1. Chromosomal DNA Extraction 

The DNA extraction was carried out using bioneer 

bacterial extraction kits (Genomic DNA extraction kits) 

protocols - ―Bioneer accuprep genomic DNA extraction 

kit (K-3032). 

Standard inoculumn (a density of 1x10
8
 cells/ml) of 

Staphylococcus aureus were prepared from 24 hours 

broth culture. 

Two millilitre (2 ml) of the prepared standard 

inoculumn was transfered to 5 ml sterile eppendorf tube 

and centrifuged for 5 min at 10,000 rpm. The supernatant 

was carefully discarded without disturbing the pellet. 

Another two millilitres (2 ml) of the standard inoculumn 

added and centrifuged at 10,000 rpm for 5 min., followed 

by carefully discarding the supernatant, and repeated 

once again to obtained more quantity of DNA. 

The pellets obtained was resuspended in 200 µl of 

phosphate buffer saline (PBS) in the eppendorf tube. 

Twenty microlitres (20 µl) of proteinase k was added 

to the tube containing the pellet in PBS, followed by 

addition of 10µl of RNase, then mixed thoroughly by 

vortexing and incubated at room temperature. 

Two hundred microlitres (200 µl) of GB buffer (lysis 

buffer) was added to the sample and mixed by vortexing, 

followed by incubation at 60°C for 10 minutes using 

heating block. 

Four hundred micro litres (400 µl) of absolute ethanol 

(Biological grade) was added and mixed well by 

pipetting, followed by careful transfered of the lysate into 

the upper reservoir of the binding or absorption column 

(fitted in the collection tube) without wetting the rim. The 

tube was closed and centrifuged at 8,000 rpm for 1 min. 

followed by discarding the solution from the collection 

tube and then resused the collection tube. 

Five hundred micro litres (500 µl) of W2 buffer was 

added without wetting the rim, followed by closing the 

tube and then centrifuged at 8,000 rpm for 1 minute. The 

solution from the collection tube was discarded and then 

reused the collection tube. 

The sample was centrifuged once more at 13,000 rpm 

for 1 minute to completely removed ethanol, followed by 

checking to ensured that there were no droplets clinging to 

the bottom of the binding column tube. The binding 

column tube was transfered to new 1.5ml tube for elution 

and 100 µl of EA buffer (elution buffer) was added on to 

the binding column tube and then kept at room 

temperature (15-25°C) for 1 minute. 

2.5.2. Polymerase Chain Reaction (PCR) - 

Accupower Hotstart PCR Premix (Bioneer) 

Twenty microlitres (20µl) reaction PCR set - up was 

prepared by adding; 16ul dH2O, 1µl forward primer - 

GGACTACAGGGTATCTAAT 16S (RIBOSE-1), 1µl 

reverse primer - AGAGTTTGATCCTGG 16S (RIBOSE-

2), and 2ul 

template DNA. PCR amplification reaction was 

performed using PTC 100 thermal cycler with Pre- 

denaturation at 95°C for 5 minutes, denaturation at 94°C 

for 1 minute, primer annealing at 54°C 1 minute, extension 

at 72°C 1 minute for 25 cycles, and final extension at 

72°C 5 minutes. The PCR products were separated by 

electrophoresis in 1.5% agarose gel for 35 minutes at 125 

volt and then visualized the gel DNA bands using UV 

lightbox/ gel imaging system (Biorad). Amplified PCR 

products were sequence and the nucleotides sequences of 

the 16SrRNA genes were searched for sequences 

similarities using online BLASTn. 

2.6. Antimicrobial Susceptibility Tests Using 

Selected Conventional Antimicrobial Agents 

Used for Treatments of Wound Infections 

Antimicrobial susceptibility test against Staphylococcus 

aureus isolates was carried out using Kirby-Buaer disc 

diffusion techniques described by Arora [17]. A loopful of 

24 hours growth culture of each isolate in nutrient broth 

was suspended in 10ml sterile distilled water and then 



    ISSN : 2693 6356 

2022 | Vol 5 | Issue 3 

 
diluted in steps of 1:10 to give turbidity equivalent to the 

0.5 McFarland standards (a density of 1x10
8
 cells/ml) 

before inoculation. Sterile cotton wool swabs were dipped 

in the suspensions adjusted to 1x10
8
 cells/ml, the excess 

fluid was removed by pressing and rotating the swabs 

against the wall of the tubes, and then streaked on the 

surface of Muller Hinton agar plates. The inoculated plates 

were allowed to dry for about 5 minutes. Using disc 

dispenser, single disc Gram positive antibiotics (Oxoid); 

Gentamycin (10µg), Amoxicillin- Clavulanic acid (30µg), 

Nalidixic acid (30µg), Kanamycin (30µg), Ciprofloxacin 

(5µg), Vancomycin (30µg), Ampicillin (10µg), Oxacillin 

(1µg), Chloramphenicol (30µg), Imipenem (10µg), 

Cefoxitin (30µg), and Sulphamethaxole (25µg) were 

dispensed on inoculated plates of Staphylococcus aureus. 

After 30 minutes of applying the discs, the plates were 

then incubated aerobically at 37°C for 24 hours in an 

inverted position. Diameter of zone of growth inhibition 

were measured using a transparent metric ruler and the 

results were interpreted as either susceptible, 

intermediate, or resistant according to Clinical and 

Laboratory Standard Institute (CLSI) guidelines [18]. 

2.7. Collection and Authentication of 

Terminalia avicennioides Plant 

Materials 

Fresh Terminalia avicennioides plant’s parts was 

collected and transported for identification at the 

Herbarium Unit of Department of Biological Science, 

Faculty of Life Sciences, Ahmadu Bello University 

Zaria, Nigeria; where the voucher number of the plant 

was obtained (900239). Fresh Terminalia avicennioides 

plant’s parts was collected after the authentication of the 

plant in large quantity and cut into small pieces and dried 

under shade at 30°C in a clean laboratory cabinet. The 

dried plant materials wasl first pounded in a mortar, 

followed by dry-milling with an electric blender and then 

sieved to obtained fine powder using 20µm mesh size 

sieve. 

2.8. Preparation of Plant Extracts 

Water, acetone and ethanol were used as the extracting 

solvents. Twenty-five gram (25g) of the processed fine 

powder sample of plant was soaked in 250ml of ethanol 

in clean sterile 500ml conical flask and then covered the 

mouth of the flask with non-absorbent cotton wool 

followed by wrapping with aluminum foil paper. The 

flask was then agitated at 80 rpm for about 48 hours at 

28±2°C using shaking incubator. The content was 

filtered first using clean muslin cloths, followed by 

Whatman’s No. 1 filter paper. The filtrate was then 

evaporated using rotary evaporator to concentrate the 

extracts at 37°C. The same procedure was repeated with 

water and acetone as the extraction solvents. 

2.9. Qualitative and Quantitative Phytochemical 

Screening 

The extracts were subjected to qualitative 

phytochemical tests to determine the presence of 

saponins, tannins, phenolic compounds, anthraquinones, 

cardiac glycosides, alkaloids, and flavonoids, using 

standard procedures described by Trease and Evans, 

Harborne, and Sofowara [19-21]. The quantitative 

Phytochemical Test was also carried out for 

detection of the amount of total Phenol, Flavonoids, 

Alkaloids, Saponins, Tannins, and terpenoids according to 

standard procedures described by; Harborne, AOAC, 

Chang et al., Edeoga et al. and Oloyed [20, 22-24]. 

2.10. In vitro Determination of Antimicrobial Activity 

of the Terminalia avicennioides Extracts Against 

Multi Drug Resistant Staphylococcus aureus 

Isolates 

2.10.1. Determination of Antimicrobial Potency 

The antimicrobial potency of the plants extracts and 

AgNPs against all the multi drug resistant Staphylococcus 

aureus isolates was determined using a spread-plate and 

agar-well diffusion method according to Ochai and 

Kolhatkar [16], and Cheesbrough [14]. Zero-point eight 

grams of the extracts of Terminalia avicennioides was 

reconstituted in 2ml of 10% Dimethyl Sulfoxide (DMSO) 

in water to get a concentration of 400mg/ml. 200mg/ml, 

100mg/ml, 50mg/ml, and 25mg/ml concentrations were 

made from the initial concentration using a standard 

dilution method. Twenty millilitres (20 ml) of Sterile 

Muller-Hinton agar was poured into each of the petri plate 

and allowed to solidify on the bench. An overnight broth 

cultures of each pure isolate was prepared, and 0.1ml of 

the culture broth was added to 19.9ml steriled distilled 

water, then adjusted by comparing with 0.5 Mcfarland 

turbidity standard (density of 1.0×10
8
 cells/ml) against a 

light background. Steriled cotton wool swab was dipped 

into the suspension, remove the excess fluid by pressing 

and rotating the swabs against the wall of the tubes and 

then streaked uniformly on the surface of Muller- Hinton 

culture plates. The inoculated plate was allowed to dry for 

5minutes. Six millimetres (6mm) diameter cork borer was 

used to make wells on the inoculated culture plates and 

0.2ml each of the reconstituted extracts concentrations was 

then loaded into the wells using sterile micropipettes. The 

plates were kept on the laboratory bench for 2 hours to 

allow the loaded extracts diffused into the culture 

medium. The plates were then incubated aerobically for 

24 hours at 37°C. This was repeated using 1mg/ml of 

ciprofloxacin as positive control; and also 2% dimethyl 

sulphur oxide (DMSO) as negative controls. Zones of 

growth inhibition form around the wells were measured 

with a transparent meter rule and the results recorded in 

millimeter (mm). The antimicrobial activity was 

expressed as the average diameter of the zones of growth 

inhibition (mm). 

2.10.2. Determination of Minimum Inhibitory 

Conentration (MIC) 

The concentrations that showed antimicrobial activity 

from the potency test were selected for the determination 

of the minimum inhibitory concentrations of the solvents 

extracts against the multi drug resistant Staphylococcus 

aureus isolates. Zero-poit eight grams of the extract of 

Terminalia avicennioides was reconstituted in 4 ml of 

10% Dimethyl Sulfoxide (DMSO) in water to get a 

concentration of 200mg/ml. 100mg/ml, 50mg/ml, and 

25mg/ml were prepared from the stock solution using 

Muller-Hinton broth as the diluent. An overnight broth 



cultures of each pure isolate was 
prepared and 0.1ml of the broth culture was added to 

19.9ml steriled distilled water, then adjust by comparing 

with 0.5 Mcfarland turbidity standard (density of 1.0×10
8
 

cells/ml) in light background. Zero-point two millilitres 

each of the 10
8
cfu/ml isolate suspension was transfered 

to 2ml of each selected solvent extract concentration in 

tubes and gently mixed by shaking the tubes. The tubes 

were then incubated aerobically at 37°C for 24 hours. 

The lowest concentrations of the extracts which showed 

no visible growth was recorded as the minimum 

inhibitory concentrations of the extracts. 

2.10.3. Determination of the Minimum 

Bactericidal Conentration (MBC) 

For each of the test tubes in the MIC that showed no 

visible growth, a loopful of the broth cultures were 

collected from those tubes and streaked on sterile 

antibiotic free nutrient agar plates. The plates were 

incubated at 37°C for 24 hours. The concentrations at 

which no growth was observed were noted and recorded 

as the minimum bactericidal concentration (MBC) [25]. 

2.11. Data Analysis 

Analysis of Variance (one way-ANOVA), Duncan 

multiple test, and independent T-test using SPSS version 

23, were used for the data analyses. 

 

3. Results 

3.1. Morphological and Biochemical 

Characteristics of Presumptive Staphylococcus 

aureus 

Presumptive Staphylococcu aureus colonies showed by 

table 1 appeared completely yellowish in colour with 

raised, circular and smooth edges on Manitol Salt agar 

(MSA). On Baired Parker agar, the colonies appeared 

black with shining characteristics and lytic edges. On 

blood agar, the colonies showed complete lysis of blood 

cells surrounding the colonies- characteristics of beta-

hymolysis. Gram stains cell appeared purple/bluewish in 

colour (Gram-positive characteristics) and cocci in shape, 

arranged inclusters (grape-like) under microscopic 

examination. The biochemical characteristics showed 

that the isolates are not motile, but catalase positive, 

coagulate positive, indole negative, methyl red positive, 

Voges- Proskauer positive, citrate utilization positive, beta-

heamolytic, lactose utilization negative, mannitol 

utilization positive and sucrose utilization negative. 

3.2. Molecular Characteristics of Staphylococcus 

aureus Isolates 

Figure 1 showed the Gel electrophoresis of amplified 

PCR 16SrRNA genes bands of Staphylococcus aureus 

isolates respectively at 800bp of the 100 bp plus DNA 

marker. The sequences BLAST results (table 2) of the 

presumptive Staphylococcus aureus isolates; S1, S2 and 

S3 16SrRNA genes revealed the percentage identity and 

similarity of these isolates from the GeneBank database 

as 76.87%, 91.64% and 86.94% respectively, confirming 

the identity of these isolates 

 Staphylococcus aureus strains. 
 

Table 1. Morphological and Biochemical Characteristics of Presumptive Staphylococcus aureus Isolates. 

 
Probable 

Organism 
Biochemical Characteristics Morphological Characteristics 

Isolate 

Idenfication Code 



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2022 | Vol 5 | Issue 3 

 
 

 

 

 

 

 

 

 

 

 
 

Keys: + = positive, - = negative, DR = dressing room wound isolate, FSW = female surgical ward wound isolate, and MSW = male surgical ward isolate. 

 
Table 2. BLAST Characteristics of Staphylococcus aureus Strains. 

 

S/N 
Sample 

Code 
Organism 

Sequence 

Searched Gene 

Total 

Scores 

Identity and 

Similarity (%) 
E-Value 

Query 

cover (%) 

Sequence Searched 

Accession No 

1. S1 Staphylococcus aureus 16SrRNA 134 76.87 8e-29 44 LT6805131 

2. S2 Staphylococcus aureus 16SrRNA 878 91.64 0.0 99 LC429749.1 

3. S3 Staphylococcus aureus 16SrRNA 360 86.94 9e-94 43 LC57519.1 

Key: S1 = DR12, S2 = FSW1, S3 = DR11, DR = dressing room wound isolate, FSW = female surgical ward wound isolate, and MSW = male surgical ward 

isolate. 
 

Figure 1. Gel electrophoresis of amplified PCR 16SrRNA genes bands of Staphylococcus aureus isolates at 800bp of the 100 bp plus DNA marker. 

 

Key: M = 100bp DNA marker, S = Staphylococcus aureus, bp = base pair, - Ve = Negative Control 12, 

S2 = FSW1, S3 = DR11 

 
3.3. Antimicrobial Activity of Selected Conventional Antibiotics Against Staphylococcus aureus Strains 

Tables 3 and 4 showed that all Staphylococcus aureus strains are multi-drug resistant isolates. Out of eleven Staphylococcu 

aureus isolates screened using twelveselected conventional antibiotics, 2 (18.18%) were resistant to   gentamycin,   3   

(27.27%)   resistant   to   kanamycin,   5 
(45.45%) resistant to ciprofloxacin, 7 (63.64%) resistant to chloramphenicol and vancomycin, 10 (90.91%) resistant to 

amoxicillin-clavulanic acid and sulphamethoxazole, and 11 (100.00%) resistant to ceftazidime, ampicillin, oxacillin and 

cefoxitin. All 11 (100.00%) isolates were sensitive to imipenem. The resistant pattern of Staphylococcu aureus isolates 

showed by table 4 indicated that four isolates (DR19, DR21, FSW1 and FSW6) were resistant each to 7 (58.33%) antibiotics 

used, five isolates (DR3, DR5, DR11, MSW3 and MSW4) were resistant each to 8 (66.64%) antibiotics used, and two isolates 

(DR12, and MSW2) were resistant each to 9 (75.00%) antibiotics. According to the results; imipenem, gentamycin and 

kanamycin were the most effective antibiotics against all the Staphylococcus aureus strains. 

Table 3. Antimicrobial activity of Selected Conventional Antibiotics against Staphylococcus aureus strains. 
 

Antibiotics Strength 
   Staphylococcus aureus  

Sensitive 

(n =11) n(%)  

Intermediate 

 
Resistant 

Gentamycin 10 µg 9 (81.18) 0 (0.00) 2 (18.18) 

Amoxicillin-Clavulanic acid 30 µg 1 (9.09) 0 (0.00) 10 (90.91) 

Kanamycin 30 µg 7 (63.64) 1 (9.09) 10 (90.91) 

Ciprofloxacin 5 µg 4 (36.36) 2 (18.18) 5 (45.45) 

Vancomycin 30 µg 4 (36.36) 0 (0.00) 7 (63.64) 



Ceftazidine 30 µg 0 (0.00) 0 (0.00) 11 (100.00) 

Ampicillin 10 µg 0 (0.00) 0 (0.00) 11 (100.00) 

Oxacillin 1 µg 0 (0.00) 0 (0.00) 11 (100.00) 

Chloramphenicol 30 µg 4 (36.36) 0 (0.00) 7 (63.64) 

Imipenem 10 µg 11 (0.00) 0 (0.00) 0 (0.00) 

Cefoxitin 30 µg 0 (0.00) 0 (0.00) 11 (100.00) 

Sulphamethoxazole 25 µg 1 (9.09) 0 (0.00) 10 (90.91) 

 
Table 4. Susceptibility Profile of Staphylococcus aureus Strains against selected antibiotics. 

 

Staphylococcus aureus 
   Conventional Antibiotics (n =12) n(%)  

Sensitive Intermediate 

 
Resistant 

DR3 4 (33.33) 0 (0.00) 8 (66.67) 

DR5 4 (33.33) 0 (0.00) 8 (66.64) 

DR11 3 (25.00) 1 (8.33) 8 (66.64) 

DR 12 2 (16.67) 1 (8.33) 9 (75.00) 

DR19 5 (41.67) 0 (0.00) 7 (58.33) 

DR21 5 (41.67) 0 (0.00) 7 (58.33) 

FSW1 4 (33.33) 1 (8.33) 7 (58.33) 

FSW6 5 (41.67) 0 (0.00) 9 (75.00) 

MSW2 3 (25.00) 0 (0.00) 9 (75.00) 

MSW3 4 (33.33) 0 (0.00) 8 (66.64) 

MSW4 4 (33.33) 0 (0.00) 8 (66.64) 

Key:, DR = dressing room wound isolate, FSW = female surgical ward wound isolate, and MSW = male surgical ward isolate. 

 
Table 5. Percentage Extracts Yield of Terminalia avicennioides. 

 

S/N Extract Category Mean±SD Extract Yield (%) P-value at α = 0.05 Comment 

 Leave, Stem and Root Bark Extracts   The percentage extracts yield based on plant 
 Leaves 5.19 ±1.61b 0.0052 extracts showed significant difference. 

1 Stem bark 15.98 ± 3.95a (P< 0.05) Stem and root bark extract showed higher 
 Root bark 13.28 ± 3.75a  percentage yield compared to leave extracts 
 Acetone, Ethanol and Aqueous Extracts    

 Acetone 11.95±6.90a 0.5209 Percentage extracts yield based on extracting 
2 

Ethanol 14.09 ± 6.42a (P> 0.05) solvents showed no significant difference. 
 Aqueous 8.40 ±3.66a   

 
Table 6. Phytochemical Characteristics of Root Barks, Stem Barks and Leave Exracts of Terminelia avecenoides. 

 

Terminelia 

S/No  avecenoides 

Plant Part 

Type of 

Solvent 

Extract 

  Phytochemical Characteristics  

Alkaloids  Flavonoids Tannins 

 

Saponins 

 

Cardiac glycosides 

 

Phenols 

 

Anthraquinones 

 

Terpenoids 

1. Root Barks Ethanol + + + + + + - + 
 Acetone - + + + - + - - 
 Aqueous - + + + + + - - 

2. Stem Barks Ethanol + + + + + + - - 
 Acetone + + + + + + - + 
 Aqueous + + + + + + - - 

3. Leaves Ethanol - + + + - + - + 
 Acetone - + + + - + - + 
 Ethanol - + + + + + - + 

Key: + = Positive; - = Negative. 

3.4. Percentage Extract Yield of Terminalia avicennoides 

The extracts from Terminalia avicennoides were obtained from dried processed powdered of stem bark, root bark and leaves 

using three extracting solvent; ethanol, acetone and water (Table 5). The percentage extracts yield based on plant parts showed 

significant difference (P< 0.05). The percentage extractsyields ranged from 5.19±1.61– 15.98 ±3.95%., with stem bark extracts 

having high percentage yied (15.98 ±3.94%). Based on extracting solvents, percentage extracts yield showed no significant 

difference (P > 0.05) and percentage extracts yield ranged from 8.40 ±3.66 - 14.09 ± 6.42%, with ethanol stem bark having high 

percentage yield (14.09±6.42%). 
 

Table 7. Quantitative Phytochemical Analysis of Root Bark, Stem Bark, and Leave Extracts of Terminelia avecenoides. 
 

T. Phenols (mg/100g) Flavonoids (mg/100g) Tannins (mg/100g) Terpenoids (mg/100g) Saponins (µg/g) Alkaloids (mg/100g) 

EET L 176.00 ± 10.50 84.00 ± 3.30 120.00± 2.60 887.00 ± 4.20 24.31 ± 0.76 129.52 ± 1.96 

EET RB 273.00 ± 10.70 84.40 ± 1.30 102.00 ± 1.50 68.00 ± 1.20 47.27 ± 1.72 298.33 ± 1.12 

EET SB 123.00 ± 20.80 88.00 ± 4.20 89.00 ± 11.00 Not Detected 45.93 ± 2.20 122.48 ± 4.96 

AQET L 362.00 ± 20.10 77.00 ± 8.10 104.00± 1.60 35.00 ± 1.70 19.90 ± 1.02 312.43 ± 0.96 

AQET RB 34.00 ± 10.12 100.00 ± 13.00 83.00 ± 3.70 Not Detected 37.35 ± 3.14 236.40 ± 0.48 

AQET SB 540.00± 20.10 111.00 ± 10.00 112.00 ± 10.00 Not Detected 22.72± 1.31 275.28 ±1.48 

AET L 2331.00 ± 23.00 106.00 ± 4.30 114.00 ± 3.50 388.00 ± 3.00 37.76 ± 3.20 131.73 ± 1.21 

AET RB 96.00 ± 10.10 104.00± 13.00 91.00 ± 3.60 Not Detected 37.79± 2.30 127.60 ± 0.72 

AET SB 1660.00 ± 12.00 104.00 ± 17.00 108.00± 3.50 56.00 ± 3.00 45.22 ± 4.21 323.82 ± 3.12 

Key: 

EET L: Ethanol Extract Terminalia Leaves; EET RB: Ethanol Extract Terminalia Root Back; 



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2022 | Vol 5 | Issue 3 

 
EET SB: Ethanol Extract Terminalia Stem Back; QET L: Aquoues Extract Terminalia Leaves; 

AQET RB: Aquoues Extract Terminalia Root Back; AQET SB: Aquoues Extract Terminalia Stem Back; 

AET L: Acetone Extract Terminalia Leaves; AET RB: Acetone Extract Terminalia Root Back; 

AET SB: Acetone Extract Terminalia Stem Back. 
 

 
3.5. Qualitative and Quantitative Phytochemical Characteristics of Root Barks, Stem Bark and Leaves Extract of 

Terminalia avicennoides 

Table 6 showed the presence of flavonoids, tannins, saponins and phenol in all the root bark, stem bark and leaves extracts 

obtained using both ethanol, acetone and water solvents. Alkaloids was detected only in ethanolic extracts of root bark, stem 

bark and also acetone aqueous stem barkextracts. Cardiac glycoside was detected only in all stem bark, ethanolic and 

aqueous root bark extracts and also ethanolic leaves extracts. Terpenoids was present in all leave extracts acetone stem bark and 

ethanol root bark extracts. Anithroquinone was not detected in all the extracts. The quantitative analysis (Table 7) showed that 

the extracts generally had higher phenol content (2331-34mg/100g), followed by terpenoids (887-35mg/100g), and then 

Saponins (47.27-22.72 µg/g) as the lowest. 



 

 

Figure 2. Showing zone of growth inhibition of Terminalia aviceniodes extract. 

Table 8. Antimicrobial Activity of Terminalia avicennioides Extracts Against Multidrug Resistant Staphylococcus aureus Strains. 
 

Organism Variably 
Mean ± SD zone of 

growth inhibition (mm) 

P-value 

at α = 0.05 
interpretation 

 Leaves, Stem and Root Bark Activity    

 AETL 16.45 ± 11.38c 0.0003  

 EETL 18.56 ± 11.63bc (P < 0.05)  

 AQTL 17.02± 10.92c   

 AETSB 23.38 ± 5.98ab  Generally, zone of growth inhibition showed 

 
Staphylococcus 

EETSB 

AQTSB 

16.28 ± 10.45c 

20.86 ± 6.38abc 
- 

significant difference. 

aureus Strains AETRB 23.81 ± 6.69a   

(DR3, DR5, 

DR11, DR12, 

EETRB 

AQTRB 

23.25 ± 6.51a 

21.00 ± 6.99abc 
- 

 

DR19, DR21, Plant Parts Extracts Activity   There was a significant difference between leave, 
FSW1, FSW6, Leaves 17.34 ± 11.24b 0.0003 stem and root bark activity. Stem and root bark 
MSW2, MSW4) Stem bark 20.17 ± 8.33a (P < 0.05) extracts showed larger zone of growth inhibition 

 Root bark 22.69 ± 6.77a  compered to leave extracts 
 Concentration (mg/ml)   There was significant difference between the zone of 

 200 

100 

50 

25.21 ± 3.45a 

24.71 ± 5.34a 

23.15 ± 7.00ab 

0.0364 

(P < 0.05) 

growth inhibition for four concentrations tested 

against the organisms. 200mg/ml, 100mg/ml 

showed larger zone of growth inhibition compared 
 25 21.15 ± 4.37a  to 50mg/ml and 25mg/ml activity 

Key:: DR = dressing room wound isolate, FSW = female surgical ward wound isolate, and MSW = male surgical ward isolate, EETL = ethanolTerminalia 

avicennioides Leave extract, AETL =acetoneTerminalia avicennioides Leave extract, AQTL =AquoeusTerminalia avicennioides Leave extract, 

EETSB=ethanolTerminalia avicennioides stem bark extract, AETSB acetoneTerminalia avicennioides stem bark extract, AQTSB = aqueousTerminalia 

avicennioides stem bark extract, EETRB = ethanolTerminalia avicennioides root bark extract, AETRB = acetoneTerminalia avicennioides root bark extract, 

and AQTRB =aqueousTerminalia avicennioides root bark extract. 

 

Table 9. Antimicrobial Activity of Terminalia avicennioides Extracts Against Multidrug Resistant Staphylococcus aureus Strains. 
 

Organism Variable 
Mean ± SD zone of 

growth inhibition (mm) 

P-value 

at α = 0.05 
interpretation 

Leave Extracts Activity    

AETL 16.45 ± 11.39a 0.7431 There was no significant different between the 

EETL 18.56 ± 11.64a (P > 0.05) activity of acetone, ethanol and aqueous extracts 

AQTL 17.02 ± 10.92a   

Concentration (mg/ml)   There was significant different between activity 

200 21.81 ± 12.93a 0.0298 at 200mg/ml and 100ml, and 50mg/ml and 

100 19.00 ± 11.63a (P < 0.05) 25mg/ml. Higher activity was recorded at 

50 15.81 ± 9.89ab  200mg/ml and 100mg/ml compare to 50mg/ml 

25 12.75 ± 8.68a  and 25mg/ml 

Stem Bark Extract Activity   There was significant different between acetone, 

AETSB 23.38 ± 5.98a 0.0019 aqueous extract, and ethanol extract activity. 
DR3, DR5, DR11, EETSB 16.28 ± 10.45b (P < 0.05) Acetone and aqueous extracts showed larger 
DR12, 

AQTSB 20.85 ± 6.38a  zones compared to ethanol extract activity. 
DR19, DR21, 

Concentration (mg/ml)   Three was a significant differennce between the 
FSW1, FSW6, 200 24.63 ± 8.39a  extracts activity for all the concentrations, with 
MSW2, MSW4, 100 22.00 ± 7.67ab 0.0003 larger zone of growth inhibition at 200mg/ml, 



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2022 | Vol 5 | Issue 3 

 
MSW4 50 18.88 ± 7.23bc 

(p < 0.05) 
followed by 100mg/ml, 50mg/ml and then 

25 15.19 ± 7.27c  25mg/ml 

Root Bark Extracts Activity    

AETRB 23.81 ± 6.69a 0.2146 There was no significant difference between both 

EETRB 23.25 ± 6.51a (p < 0.05) acetone, ethanol, and aqueous extracts 

AQTRB 21.00 ± 6.99a   

Concentration (mg/ml)   Three was a significant difference between the 

200 28.29 ±4.14a 0.0001 extracts activity for all the concentrations, with 

100 25.10 ± 4.49b (p < 0.05) larger zone of growth inhibition at 200mg/ml, 

50 20.96 ± 5.53c  followed by 100mg/ml, 50mg/ml and then 

25 16.39 ± 6.17d  25mg/ml 

Key:: DR = dressing room wound isolate, FSW = female surgical ward wound isolate, and MSW = male surgical ward isolate, EETL = ethanolTerminalia 

avicennioides Leave extract, AETL =acetoneTerminalia avicennioides Leave extract, AQTL =AquoeusTerminalia avicennioides Leave extract, 

EETSB=ethanolTerminalia avicennioides stem bark extract, AETSB acetoneTerminalia avicennioides stem bark extract, AQTSB = aqueousTerminalia 

avicennioides stem bark extract, EETRB = ethanolTerminalia avicennioides root bark extract, AETRB = acetoneTerminalia avicennioides root bark extract, 

and AQTRB =aqueousTerminalia avicennioides root bark extract. 

 

3.6. Antimicrobial Activity of Terminalia avicennoides Extracts Against Multi drug Resistant Staphylococcus aureus Strains 

Figure 2 showed the zone of growth inhibition produced by the activity of Terminalia avicennioides extracts. Antimicrobial 

activity of Terminalia avicennioides extacts against multidrug resistant Staphylococcus aureus isolate result in tables 8 and 9 

showed in vitro activity of theacetone, ethanol and aqueous extracts of stem bark, root bark and leave extracts as zone 

of growth inhibition in millimeter for four varying concentrations: 200 mg/ml, 100 mg/ml, 50 mg/ml and 25 mg/ml. 

The zone of growth inhibition ranged from 16.28±10.45 – 23.81±6.69 mm and showed significant difference (P < 

0.05), with acetone root and stem bark, ethanol root bark and aqueous leave extracts showing larger zone of growth 

inhibition. 
 

Table 10. Minimum inhibitory Concentration (MIC) of Terminalia avicennioides Extracts against Multidrug Resistant Staphylococcus aureus strains. 
 

Organism Variable 
Mean ± SD MIC 

(mg/ml) 

P-value 

at α = 0.05 
Interpretion 

 Leave, Stem and Root Bark    

 
 

Generally, there was no significant difference between 

the MIC for all the extracts irrespective of the parts of 

the plant and type of the extracting solvents extracts 

tested against all the multidrug resistant Staphylococcus 

aureus strains. 

 

 

 

 
No Significant difference between the MIC for the leave, 

stem and root bark extracts activity against all the 

bacterial strains 

 
No Significant difference between the MIC for the 

acetone, ethanol and aqueous leave extracts activity 

against all the bacterial strains 

 
No Significant difference between the MIC for the 

acetone, ethanol and aqueous stem bark extracts activity 

against all the bacterial strains 

 

Significant difference between the MIC for the acetone, 

ethanol and aqueous stem bark extracts activity against 

all the bacterial strains 

 Extracts Activity   

 AETL 31.25 ± 22.16a  

 EETL 37.50 ± 32.73a  

 AQTL 

AETSB 

EETSB 

43.75 ± 32.04a 

56.25 ± 29.12a 

43.75 ± 39.52a 

0.7804 

(P > 0.05) 

 AQTSB 53.12 ± 31.16a  

 AETRB 43.75 ± 25.87a  

 EETRB 43.75 ± 11.57a  

 AQTRB 53.12 ± 31.16a  

Staphylococcus Plant Parts Extracts Activity   

aureus strains (DR3, 

DR5, DR11, DR12, 

DR19, DR21, FSW1, 

FSW6, MSW2, 

MSW3, MSW4) 

Leaves 

Stem bark 

Root bark 

Leave Extracts Activity 

AETL 

37.50 ± 28.55a 

51.04 ±32.54a 

46.87 ±23.67a 

 
31.25 ± 22.16a 

0.2480 

(P > 0.05) 

 

 

0.7005 

 EETL 37.50 ± 32.73a (P > 0.05) 

 AETL 43.75 ± 32.04a  

 Stem Bark Extracts Activity   

 AETSB 56.25 ± 29.12a 0.7437 

 EETSB 43.75± 39.53a (P > 0.05) 

 AQTSB 53.13 ± 31.16a  

 Root Bark Extracts Activity   

 AETRB 43.75 ± 25.87a 0.6778 

 EETRB 43.75 ± 11.57a (P > 0.05) 

 AQTRB 53.12 ± 31.16a  

Key:: DR = dressing room wound isolate, FSW = female surgical ward wound isolate, and MSW = male surgical ward isolate, EETL = ethanolTerminalia 

avicennioides Leave extract, AETL =acetoneTerminalia avicennioides Leave extract, AQTL =AquoeusTerminalia avicennioides Leave extract, 

EETSB=ethanolTerminalia avicennioides stem bark extract, AETSB acetoneTerminalia avicennioides stem bark extract, AQTSB = aqueousTerminalia 

avicennioides stem bark extract, EETRB = ethanolTerminalia avicennioides root bark extract, AETRB = acetoneTerminalia avicennioides root bark extract, 

and AQTRB =aqueousTerminalia avicennioides root bark extract. 

 



3.7. Minimum Inhibitory Concentration (MIC) of Terminalia avicenode Extracts Against Multi Drug Resistant 

Staphylococcus aureus Strains 

As presented in table 10 the MIC of leave, stem and root bark extracts for all types of solvent extracts tested against multi 

drug resistant Staphylococcus aureus isolate strains ranged from 56.25±29.12 – 31.25±22.16 mg/ml and showed no significant 

difference (P> 0.05). However, acetone extracts showed higher MIC value of 31.25±22.16 mg/ml, and acetone stem bark 

extracts showed the lower MIC values of 56.25±29.12 mg/ml. 

3.8. Minimum Bactericidal Concentration (MBC) of Terminalia avicennioides Extracts Against Multidrug Resistant 

Staphylococcus aureus Strains 

As presented in table 11, the MBC of leave, stem and root bark extracts for all types of solvent extracts tested against multi 

drug resistant Staphylococcus aureus isolate strains ranged from 175.00±46.29 – 68.75±45.81 mg/ml and showed no significant 

difference (P> 0.05). However, acetone leav extracts showed higher MBC (68.75±45.81 mg/ml), and aqueous stem bark 

extracts showed the lower MBC values of 175.00±46.29 mg/ml. 

Table 11. Minimum Bactericidal Concentration (MBC) of Terminalia avicennioides Extracts against Multidrug Resistant Staphylococcus aureus. 
 

Organism Variable 
Mean ± SD MBC 

(mg/ml) 

P-value 

at α = 0.05 
Interpretation 

 Leave, Stem and Root Bark Extracts Activity    

 
The MBC showed significant 

difference. However, acetone stem 

bark and ethanol root bark showed 

lower MBC compared to other 

extracts. 

 

 
MBC showed no significant 

difference. However, leave extracts 

showed higher MBC compared to 

stem and root bark extracts. 

MBC showed no significant 

different. However, acetone extracts 

showed higher MBC compared to 

stem and root bark extracts. 

 
MBC showed no significant 

different. 

 
 

MBC showed no significant 

different. 

 AETL 68.75 ±45.81b  

 EETL 75.00 ±46.29b  

 AQTL 125.00 ±88.64ab  

 AETSB 112.500 ±64.09ab 0.0388 
 EETSB 106.25 ±86.34b (P < 0.05) 
 AQTSB 175.00 ±46.29a  

 AETRB 115.00 ±45.28ab  

 EETRB 125.00 ±46.29ab  

 AQTRB 106.25 ±57.37b  

Staphylococcus aureus 

Strains (DR3, DR5, 

DR11, DR12, DR19, 

DDR21, FSW1, FSW6, 

MSW2, MSW3, MSW4) 

Plant Parts Extracts Activity 

Leaves 

Stem bark 

Root bark 

Leave Extracts Activity 

AETL 

 

89.58 ±65.90b 

131.25 ±71.95a 

112.50 ±53.67a 

 
68.75 ±45.81b 

 

0.0872 

(p > 0.05) 

 

 
0.1765 

 EETL 75.00±46.29a (p > 0.05) 
 AQTL 125.00 ±88.64a  

 Stem Bark Extracts Activity   

 AETSB 112.50 ±64.09a 0.1036 
 EETSB 106.25 ±86.34a (p > 0.05) 
 AQTSB 175.00 ±46.29a  

 Root Bark Extracts Activity   

 AETRB 115.00 ±45.28a 0.4320 
 EETRB 125.00 ±46.91a (p > 0.05) 

 AQTRB 106.25 ±57.37a  

Key: DR = dressing room wound isolate, FSW = female surgical ward wound isolate, and MSW = male surgical ward isolate, EETL = ethanolTerminalia 

avicennioides Leave extract, AETL =acetoneTerminalia avicenioides Leave extract, AQTL =AquoeusTerminalia avicennioides Leave extract, 

EETSB=ethanolTerminalia avicennioides stem bark extract, AETSB acetoneTerminalia avicennioides stem bark extract, AQTSB = aqueousTerminalia 

avicennioides stem bark extract, EETRB = ethanolTerminalia avicennioides root bark extract, AETRB = acetoneTerminalia avicennioides root bark extract, 

and AQTRB =aqueousTerminalia avicennioides root bark extract. 
 

 

4. Discussion 
Using phenotypic and genotypic methods, this research 

isolated and identified strains of Staphylococcus 

aureus from individuals with wound infections. 

Staphylococcus aureus colonies on mannitol salt agar 

(MSA) were found to be yellow, flat, and moderately 

shaped, according to cultural morphology of 

phenotypic identification. Fitzgerald said that the 

fermentation of mannitol salt, leading to the creation of 

acid, is responsible for the formation of yellow 

colonies on MSA [26]. Colonies of Staphylococcus 

aureus appeared dark grey-black and shiny on Baired 

Parker medium. They had an opaque halo and a 

clearing zone around them. In a study conducted by 

Silva et al., similar phenomena were observed in 

Staphylococcus aureus on Baired Parker medium. The 

authors found that the colonies' greyish-black shine is 

caused by a decrease in potassium tellurite, while the 

clear zone around each colony is the outcome of 

proteolytic activity caused by Lecithinase breaking 

down egg yolk. The opaque halo surrounding this 

clearing is thought to be caused by Lipase activity 

[27]. Under the microscope with an x100 objective 

lens, the gram stain cell showed a characteristic of 

Gram positive cocci, which is that they appeared in 

clusters like grapes. Staphylococcus aureus has a 

comparable cellular morphology, according to Tong et 

al. [28]. This organism was shown to be catalase and 

coagulase positive according to the biochemical 

characteristics.  

 



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2022 | Vol 5 | Issue 3 

 

Beta-hemolysis, a distinctive phenotypic marker for 

pathogenic Staphylococcus aureus strain identification, 

is characteristically produced on blood agar. Research 

has shown that human Staphylococcus aureus isolates 

contain both bound and free coagulase forms [16], and 

they produce the telltale beta-haemolysis when 

cultured on blood agar. One way to tell harmful 

Staphylococcus aureus strains apart from less 

dangerous ones is by looking for the presence of the 

enzyme coagulase.  

This study's phenotypic identification method 

uncovered biochemical and cultural traits associated 

with Staphylococcus aureus isolates. But because this 

ethnobotanical study had to focus on pathogen-specific 

wound infections, and because the organisms chosen 

had to be directly related to the traditional uses of the 

plant Terminalia avicennioides, it was necessary to use 

molecular identification methods to characterize 

Staphylococcus aureus. According to Vanvuuren [5], 

this is done to ensure that research can be reproduced. 

As per Prescott et al. [29], the Staphylococcus aureus 

bacteria that were found in wound infections were 

compared using molecular identification to the 

Genbank database to see how similar they were 

genetically. The gel electrophoresis of amplified PCR 

16SrRNA gene bands of Staphylococcus aureus 

isolates at 800 bp of the 100 bp plus DNA marker was 

shown by the findings of the molecular analysis in this 

research. Presumptive Staphylococcus aureus 

sequences from BLAST  

isolates; S1, S2 and S3 16SrRNA genes revealed the 

percentage identity and similarity of these isolates to 

those from the GeneBank database as 76.87%, 91.64% 

and 86.94% respectively, confirming the identity of 

these isolates as Staphylococcus aureuss trains. 

According to Prescott et al., it has been generally 

acknowledged since the 1970s that prokaryotes with 

genomes that are at least 70% homologous belong to the 

same species [5]. The percentages of identity and similarity 

shown by the sequence BLAST results for all of the 

Staphylococcus aureus strains ranged from 76.87% to -

997.67%. This lends credence to the study's conclusion that 

the isolates in question are, in fact, Staphylococcus aureus.  

All of the Staphylococcus aureus strains identified in this 

investigation were shown to be resistant to several drugs. 

All of the Staphylococcus aureus isolates tested were 

susceptible to imipenem and Gentamycin, respectively, 

suggesting that these antibiotics were the most effective 

against the bacteria (3 and 4). This implies that doctors need 

to be very cautious when prescribing imipenem to patients 

in order to prevent the organism from developing a 

resistance. The prescription of these medications to patients 

should always be based on sensitivity results. The discovery 

and its significance to public health should also be 

communicated to practitioners. Similar results were 

observed in a study of Staphylococcus aureus susceptibility 

profiles by Rashedul et al. [30], who found that 90% of the 

isolates were sensitive to imipenem and that 75% of the 

isolates were resistant to oxacillin, methicillin, 

ciprofloxacin, and tetracycline. According to research by 

Kitara et al. and Brown and Ngeno, among other sources, 

Staphylococcus aureus may develop resistance to a wide 

variety of medicines and can generate several strains that are 

resistant to these drugs [31, 32]. The authors Brown and 

Ngeno agreed that antibiotic-resistant Staphylococcus 

aureus represents an international health crisis. 

Consistent with previous research, this investigation found 

that some strains of Staphylococcus aureus were resistant to 

the antibiotic chloramphenicol [30]. In line with the findings 

of Aisha et al. [33], our investigation also documented that 

Staphylococcus aureus displayed multidrug resistance to 

ceftazidime. Also, this research found that Staphylococcus 

aureus is resistant to vancomycin, which is concerning since 

Rashedul et al. found that only 4 out of 66.63 percent of 

Staphylococcus aureus strains were sensitive to the 

antibiotic [30]. This study's findings of vancomycin 

resistance in Staphylococcus aureus isolates suggest that 

some strains of this bacterium pose a significant threat to the 

efficacy of wound infection treatments and pose an extra 

burden on healthcare systems, particularly in communities. 

Since vancomycin is still only effective against some strains 

of Staphylococcus aureus, researchers Khan et al. and 

Juayan et al. have identified VRSA as a major global health 

concern [34, 35].  

 

When it came to treating wound infections caused by 

Staphylococcus aureus, Benjamin and Christopher 

suggested tetracycline, chloramphenicol, and Gentamycin 

[36]. On a similar note,  

 

For the efficient treatment of wound infections, Bowler et 

al. suggested the following medications: imipenem, 

cefoxitin, gentamycin, and vancomycin [37]. The study's 

results showed that the most effective antibiotics, as 

advised, are Imipenem, Gentamycin, and Ciprofloxacin. 

Despite the earlier study recommending them, the other 

antibiotics tested were ineffective against the tested bacterial 

strains. Even more concerning is the fact that the 

Staphylococcus aureus strains tested here were susceptible 

to gentamycin, imipenem, and ciprofloxacin, in contrast to 

the multidrug-resistant bacteria described by Aisha et al. 

[38]. Bacterial resistance gene acquisition, mutations, 

environmental conditions, biofilm development, presence of 

beta-lactamase, efflux pump mechanism, and other factors 

could all contribute to the inconsistency. Because current 

antibiotics are ineffective against bacterial wound 

infections, this demonstrated the need for a new medication.  

Extraction solvents used in this research were ethanol, 

acetone, and water, while the plant material used was dried, 

processed powdered Terminalia avicennoides stem bark, 

root bark, and leaves. The percentage yields of the extracts 

demonstrated a significant variation (P<0.05), ranging from 

5.19±1.61 to 15.98 ±3.95%. The percentage yield of the 

extracts varied from 8.40 ± 3.66 to 14.09 ± 6.42% 

depending on the solvents used for extraction, and there was 

no significant difference (P>0.05). Possible explanations for 

the observed % yield discrepancies include the use of 

various extraction solvents. According to research by Mule 

et al., the polarity of the extracting solvent (non-polar, polar, 

or less polar) has a significant impact on the kinds of 

bioactive compounds that can be extracted from plant 

components [39]. The results showed that acetone and 

ethanol solvents produced higher percentage yields of 

extracts than water, according to this research. Since most 

active antimicrobial components are insoluble in water, a 



universal polar solvent, according to Afolayan et al. [40], it 

stands to reason that organic polar solvents like acetone and 

ethanol would produce more potent antimicrobial extracts. 

This could be because ethanol and acetone had a greater 

percentage yield from their extracts than water did in this 

investigation.  

The research found that the extracts of Terminalia 

aveicennioides included tannins, alkaloids, flavonoids, 

cardiac glycosides, phenolic compounds, terpenoids, and 

sapponins, according to the quantitative and qualitative 

phytochemical examination. None of the plant extract 

categories showed any signs of anthroquinones. Following 

terpenoids (887-35mg/100g) and saponins (47.27-22.72 

µg/g), the quantitative analysis revealed that extracts often 

exhibited a greater phenol content (2331-34mg/100g). Alaje 

et al. and Odebumin et al. both found the same thing [41, 

42]. The majority of the plant's chemical components 

include many bioactive substances, such as alkaloids, 

tannins, flavonoids, triterpenoids, phenolic compounds, 

carotenoids, steroids, and ketones, according to earlier 

research on the biochemical components of medicinal plants 

by Irshad et al. [43]. According to Radhika et al., the 

bioactive chemicals in question are very significant.  

are the alkaloids, tannins, sapponins, flavonoids and 

phenolic compounds [44]. According to Cragg and Newman, 

the presence of important phytochemical constituents is the 

bioactive bases for plant medicinal properties as these 

secondary metabolites are the chemical substances used by 

the plants for defense system and serve as bioactive 

principles for various drugs and modern therapy [45]. 

The important phytochemical constituents like steroids, 

tannins and saponins have been detected in Terminilia 

avecennoides plant parts [46], and the presence of these 

compound is known to confer antibacterial activity against 

bacteria pathogens [47]. To confer antibacterial activity of 

plant, flavonoids has been reported to be singly 

responsible for antibacterial activity associated with some 

ethnomedicinal plant [48]. It has also been reported that 

plants that are rich in tannins or phenolics compounds are 

inhibitory to wide range of bacteria, thus capable of 

confering protection against some microbial infections 

[49]. The presence of the various phytochemical 

compounds is an indication that Terminilia avecennoides 

have potent antiseptic, bactericidal and other medicinal 

properties. This is due to the fact that each of the 

compounds identified has one or more therapeutic usage 

and may be acting singly or in consortium to bring about 

cidal or static effect on the organism. Thus, the presence 

of the phytochemical compound recorded in this study 

could be responsible for in vitro antibacterial activity. 

The in vitro antimicrobial activity of the various 

Terminilia avecennoidesextracts against multi drug 

resistant Staphylococcus aureus showed zone of growth 

inhibition on the various concentrations, extracting 

solvents and parts of the plant. Antibacterial activity was 

shown by an inhibitory activity characterized by a cleared 

zone between the wells (containing the samples) and 

certain distance. Formation of inhibitory zones around the 

wells shows bacterial sensitivity to the extracts. The 

antimicrobial activity of Terminalia avicennoides extacts 

against multidrug resistant Staphylococcus aureus isolates 

showed in vitro antimicrobial activity of the acetone, 

ethanol and aqueous extracts of stem bark, root bark and 

leave extracts as zone of growth inhibition in millimeter for 

four varying concentrations: 200mg/ml, 100mg/ml, 

50mg/ml and 25mg/ml. The zone of growth inhibition 

ranged from 16.28±10.45 – 23.81±6.69 mm and showed 

significant difference (P< 0.05), with acetone root and stem 

bark, ethanol root bark and aqueous leave extracts 

showing larger zone of growth inhibition in comparison to 

others. Udgire and Pathade suggested that plant extracts 

exhibiting inhibitory zones diameter greater than or equal to 

10 mm and above against selected microbial pathogens 

should be considered to possess antimicrobial activity, 

whereas, those showing inhibitory zones greater than 20 

mm against selected microbial pathogens should be 

considered noteworthy [50]. The level of the extracts in 

vitro antibacterial activity against the multi drug resistant 

Staphylococcus aureus isolates revealed the presence of 

the important bioactive ingredients, the strength 

concentrations of these ingredients and their capacity to 

diffuse into the agar medium. In this study, thezone of 

inhibition of the extracts increases as the extract 

concentration increases, thus, the linear relationship 

between the concentrations of the extract zone of 

inhibition could be that the extracts used were able to 

diffuse into the inoculated nutrient agar. This however, 

may explain why even though there were cleared zone of 

growth inhibition for some extracts against some bacteria 

strains, there were also no detectable zone for different 

solvents extracts and different extracts concentration 

against different bacteria isolates. 

Several studies have attributed the antibacterial and 

therapeutic activities of Terminilia avecennoides extracts 

to the presence of flavonoids and a mixture of phenolic 

compounds and tannins [51]. The phenolic compounds 

are said to act as protoplasmic poison which penetrate 

and disrupt bacterial cell wall in addition to precipitation 

of cell proteins. More so, it has been confirmed that 

secondary metabolites such as alkaloids and tannins 

inhibit enzymes and protein synthesis, while glycosides 

are antidiarrhea [52]. The Terminilia avecennoides 

extracts were found to be active against the 

Staphylococcus aureus strains with greater inhibitory 

activity at concentration of 200 mg/ml and 100 mg/ml 

and this is similar to findings by Shedidi [53]. The 

present study revealed that the Terminalia avicennioides 

extracts showed potent antibacterial activity against the 

bacterial strains. This implies that the in vitro 

antimicrobial activity of the Terminilia 

avecennoidesextracts recorded in this study was due to 

availability of the plant secondary metabolites required 

for antibacterial activity. The ability of the extracts of 

Terminilia avecennoides to inhibit the growth of the 

multi drug resistant Staphylococcus aureus explains why 

it is been effectively used in folk medicine for treatment 

of wound infection. The Terminalia avicennioides is the 

most widely used plants for traditional medicinal 

purposes worldwide including wound healing. It is 

known for local used in form of; leaf and root bark 

medicine, pain killer root bark medicine, and skin and 

mucosae root bark medicine [53, 54]. Because of its 

potential antimicrobial activity, it is harvested locally and 

used for treatment of burn and wound infection. The 

pulverized leaves are used in Northern Nigeria on burns 

and bruises. In north Eastern Nigeria, the Jukun in 

Taraba state use the roots in treatment of syphilis. The 

root bark is made into a decoction along with other 

medicinal plants by the Baule of Ivory coast for severe 

jaundice and non-healing old sores. In Casamance of 

Senegal, the root bark is considered cleasing and healing 



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2022 | Vol 5 | Issue 3 

 
on refractory sores. The powdered root bark is applied 

topically to sores and ulcers and is rubbed on the gums of 

toothache in Ivory Coast. The root bark is being used for 

treatment of skin infection and separate examination of 

antimicrobial activity against Sarcina lutea, 

Staphylococcus aureus, Mycobacterium phlei, and some 

Gram positive organisms [53, 54]. It can therefore, be 

deduced from the result obtained in this study that 

Terminilia avecennoides is a source of bioactive 

compounds with potential therapeutic benefit, because it 

portrays a good inhibitory effect against the multi drug 

resistant Staphylococcus aureus. 

The Terminilia avecennoides extracts showed MIC 

values 



  

at different concentration depending on extracting solvent 

and parts of the plants. The MIC of the plant extracts 

tested against multi drug resistant Staphylococcus aureus 

isolate strains in this study ranged from 56.25±29.12 – 

31.25±22.16 mg/ml, and showed no significant difference 

(P> 0.05) with acetone extracts having higher MIC value 

of 31.25±22.16 mg/ml, and acetone stem bark extracts 

showed the lower MIC values of 56.25±29.12 mg/ml. 

Similarly, the MBC of the extracts tested against multi 

drug resistant Staphylococcus aureus isolate strains 

ranged from 175.00±46/29 – 68.75±45.81 mg/ml and 

showed no significant difference (P> 0.05) with acetone 

leave extracts having higher MBC (68.75±45.81 mg/ml), 

and aqueous stem bark extracts having the lower MBC 

values of 175.00±46.29 mg/ml. These values represent the 

in vitro bacteriostatic and bactericidal concentrations of 

these crude extracts against the multi drug resistant 

Staphylococcus aureus strains. The high concentrations of 

the secondary metabolites such as tannins, alkaloids, 

flavonoids, saponins, terpenoids, cardiac glycosides, 

among others in this plant extracts could be attributed to 

the high antimicrobial activity recorded in this study. The 

findings are indicative of the various efficacy levels of 

Terminilia avecennoides extracts that can be enhanced by 

further separation, purification and concentration of the 

bioactive compounds of the plants. 

 

5. Conclusion 
Patients both in and out of Barau Dikko Teaching Hospital 

Kaduan, Nigeria, had Staphylococcus aureus strains 

extracted from their wounds. The Staphylococcus aureus 

strains were all resistant to more than one antibiotic. Only 

four antibiotics—gentamycin, imipenem, ciprofloxacin, and 

kanamycin—were effective against the wound infections 

tested. Noteworthy antimicrobial activity against multidrug 

resistant Staphylococcus aureus strains is shown by the 

Terminalia avicennioides extracts, which include important 

phytochemical components necessary for bacteriostatic and 

bactericidal activities. The plant extracts of Terminalia 

avicennioides showed promise as a potential therapeutic 

agent for the treatment of bacterial wound infections caused 

by strains of Staphylococcus aureus that are resistant to 

several drugs. Nevertheless, rigorous research is necessary 

to identify the particular bioactive or inhibitory chemicals 

that are effective against the strains of Staphylococcus 

aureus that are resistant to multiding. 

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