AASCIT Journal of Health

2017; 4(2): 8-16

http://www.aascit.org/journal/health
ISSN: 2381-1277 (Print); ISSN: 2381-1285 (Online)

Phytochemical Screening and

Antibacterial Activity of Extracts of
Garcinia mannii and Terminalia
avicennoides on Some Oral
Bacterial Pathogens
Amuda Oladunni1, Samuel Alimi Garba1, Galadima Musa1,
Jigam Ali Audu2
1
Department of Microbiology, Federal University of Technology, Minna, Nigeria
2
Department of Biochemistry, Federal University of Technology, Minna, Nigeria

Keywords Email address

Antibacterial, amudaoamuda@gmail.com (A. Oladunni)
Phytochemicals,
Citation
G. mannii,
Amuda Oladunni, Samuel Alimi Garba, Galadima Musa, Jigam Ali Audu. Phytochemical
T. avicennoides,
Screening and Antibacterial Activity of Extracts of Garcinia mannii and Terminalia avicennoides
Amoxicillin,
on Some Oral Bacterial Pathogens. AASCIT Journal of Health. Vol. 4, No. 2, 2017, pp. 8-16.
Ampiclox
Abstract
Garcinia mannii and Terminalia avicennoides twigs are commonly used as chewing sticks
for dental hygiene among many tribal groups in Nigeria. The antibacterial activity of
Received: July 27, 2017 methanol and aqueous extracts of G. mannii and T. avicennoides were tested on some
Accepted: August 13, 2017 isolates of oral bacterial pathogens (Micrococcus luteus, Streptococcus mutans,
Published: November 13, 2017 Pseudomonas. aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae and
Streptococcus pyogenes). The percentage yield of the extract revealed 10.64% and 10.46%
for methanol and aqueous extract of G. mannii and 22.24% and 22.36% for methanol and
aqueous extract of T. avicennoides. Preliminary phytochemical screening revealed the
presence of alkaloids, phenols, tannins, flavonoids, cardiac glycosides, anthraquinones,
steroids, phlobatannins, terpenes and saponins. Quantitative phytochemical analysis
(mg/100g) showed total phenols ranged from 706.95±147.58-1017.01±18.85, flavonoids;
198.35±2.13-366.750±14.53, alkaloids; 120.31±3.15-25.54±21.96, tannins; 138.50±6.03-
224.65±1.94 and saponins; 4.81±0.00-8.28±0.28 in aqueous and methanol extracts of G.
mannii and T. avicennoides. The antibacterial test based on the diameter of clear zones of
inhibition on agar plates ranged from 6.33±0.67 – 25.00±0.57 mm at concentrations of 20,
30 and 40mg/mL in aqueous and methanol extracts of G. mannii and T. avicennoides.
These were comparable to the 5mg/mL of both standard antibiotics used (Amoxicillin and
Ampiclox) with zones of inhibition ranging from 15.67±0.33 to 28.00±1.00 on all the test
organisms. Minimum Inhibitory concentration (MIC) and Minimum Bactericidal
Concentration (MBC) of the two active extracts ranged from 0.625-20mg/mL and 1.25-
40mg/mL respectively. These results confirmed the efficacy of G. mannii and T.
avicennoides in traditional dental care. The extract could contain bioactive compounds
with potential in the formulation of new dental care medicaments.

1. Introduction
According to the World Health Organization (WHO), a medicinal plant is any plant in
which one or more of its organs or parts, contains chemical substances that can be used
for therapeutic purposes, or which are precursors for synthesis of other bioactive
metabolite. Such a plant will have all of its parts employed in the control or treatment of
9 Amuda Oladunni et al.: Phytochemical Screening and Antibacterial Activity of Extracts of Garcinia mannii and
Terminalia avicennoides on Some Oral Bacterial Pathogens

a disease condition and therefore contains chemical separate. [12]. The stick is held by one hand in a pen-like
components that are medically active. These bioactive grip and the brush end is used with an up and down or rolling
components are often referred to as phytochemicals or motion. A two finger or and a five finger grip technique
phytoconstituents and are responsible for protecting the plant maybe used. When the brushy edge is shred after being
against microbial infections or infestations by pests [1], [2], frequently used, the stick gets ineffective and it is then cut
[3]. and further chewed to form a fresh edge. In this way, it can
Medicinal plants contain biologically active components be used for few more weeks [13].
which have been employed in traditional medical practice for There are known 173 different types of trees, which can be
the treatment of human infections [4]. According to some used as chewing sticks, belonging to the families Rubiaceae,
studies carried out on some medicinal plants in Nigeria it is Rutaceae, Combretaceae, Euphorbiaceae, Meliaceae,
reported that Garcinia mannii and Terminalia avicennoides Mytaceae and Asteraceae [14]. The most popular chewing
are used in treatment of pains and oral infections. stick or fibrous sponges include Salvadorapersica and
Investigations have shown that these chewing sticks Azadirachta indica [8].
possesses antimicrobial activity against oral microbial flora The human oral cavity is maintained at a relatively
[5]. The species of trees and shrubs from which chewing comfort temperature (35°C-36°C), thus providing suitable
sticks are made is numerous, but in Southern Ghana the most conditions for a larger number of microorganisms [12]. The
popular varieties belong to the Garcinia species, including G. ecological conditions in the mouth are never stable for long
mannii, G. kola, and G. epunctata [6]. Notwithstanding the periods of time being affected by intermittent feeding and
relative abundance of literature on the benefits of chewing age. With change in the natural ecosystems such as the use of
sticks, information on their chemical and pharmacological antibiotics, contributes to the variations in microbial
components is generally scanty [6], and probably nonexistent communities [15].
in the specific case of Garcinia mannii. Oral infections are disease conditions found in the mouth.
In Africa, chewing sticks commonly used for maintaining The mouth includes not only the teeth and the gums but their
oral hygiene. The roots, stem and twigs of numerous plants supporting connective tissues, ligaments, bones, soft and hard
are employed for this purpose. Chewing sticks are palates. These conditions are generally grouped into three
recommended for oral hygiene by the World Health main categories, as diseases of the teeth, diseases of the gums
Organization [7] and some of them, or their extracts, are and oral cancers. This study is aimed at authenticating and
also used in the ethno medical treatment of oral infections scientifically verify the claims of antibacterial properties by
[8]. local marketers and users of some selected chewing sticks in
The use of chewing sticks have been documented since some parts of Nigeria.
ancient times. And have been used by the Babylonians some
7000 years ago [9]. The cleansing efficacy of chewing sticks 2. Materials and Methods
is attributed to the mechanical effects of its fibers, or release
of beneficial chemicals or a combination of both [7]. Some 2.1. Materials
Africa chewing sticks are reported to contain fluoride ions,
silicon, tannic acid and other natural plaque inhibiting 2.1.1. Collection and Identification of Plant
substances that can reduce bacterial colonization and plaque Materials
formation [10]. A significant percentage of the Nigeria Fresh stem of Garcinia mannii and Terminalia
population uses chewing sticks, mainly because they are avicennoides were collected from Okija in Anambra state and
readily available and cheap [11]. Maikunkele in Minna, Niger State both in Nigeria in
Recent interest in chewing sticks and their extracts have February, 2016. The plants were identified at National
focused on their effects on organisms that are involved in Institute of Pharmaceutical Research and Development, Idu-
oral infections. The stick is usually chewed or tapered at one Abuja, Nigeria and assigned a voucher number
end until it becomes frayed into a brush. Soaking it in water NIPRD/H/6786 and NIPRD/H/6797 for G. mannii and T.
for few hours softens the natural fibers, helping them to avicennoides respectively.
Table 1. Local names, ethno medicinal uses and locations of collection of the plants used.

Scientific name Local names Ethnomedicinal uses Part used Location

Antimalarial, antibacterial, used as
Garcinia mannii Namijin goro (H), Aku-ilu (I) stem Okija, Anambra State
preservatives, dye e.t.c
Wawan kurmi (H), erin mado (Y), Use to treat wound and skin infections, Maikunkele, Niger
Terminalia avicennoides Stem
okwe (I) dysentery, amoebiasis and diarrhea State

Keys: I – Igbo, H – Yoruba, H – Hausa

 AASCIT Journal of Health 2017; 4(2): 8-16 10

2.1.2. Sources and Characterization of 2.4. Determination of Antibacterial Activity

Bacteria Pathogens of the Extract
Samples were collected from oro-dental patients attending
General Hospital Minna, using sterile swab stick soaked with 2.4.1. Assay for Antibacterial Activity
sterile normal saline. The swab stick sample was inoculated into Agar well diffusion method was used to evaluate the
prepared nutrient broth and incubated for 6 h, so as to activate antibacterial activity of the Crude extracts [37]. Eighteen-
the bacteria. After activation in the nutrient broth, the nutrient hour culture of bacteria adjusted to 0.5 McFarland standard
broth culture was subcultured on nutrient Agar, Blood Agar, was used as inoculum on sterile Mueller Hinton agar. The
MacConkey Agar and Manitol Salt Agar so as to isolate the plate was kept on flat bench for 30 minutes to solidify. Five
bacteria. This was characterized after isolation and compared wells (4mm) deep were made in the agar using a sterile 6mm
with known existing taxa [16]. The bacteria pathogens identified diameter cork borer. Then 0.5 mL of the reconstituted extract
include: Micrococcus luteus, Streptococcus mutans, at a concentration of 20, 30 and 40 mg/mL was pipetted into
Pseudomonas aeruginosa, Klebsiella pneumoniae, the wells using micro pipette. Zero point five milliliter
Streptococcus pneumoniae and Streptococcus pyogenes. (0.5mL) each of 5mg/mL of Ampiclox and Amoxiline
solution were used as positive controls and 0.5mL of Di-
2.1.3. Ethical Consideration Methyl Sulphoxide (DMSO) as a negative control. The plates
Ethical clearance to conduct this research was sought from were allowed to stand on a flat bench for 30 min to allow
the research ethics and publication committee of the hospital. diffusion of the extract into the agar before incubation at
Informed consent was obtained for each respondent before 37°C for 24 h. Each test was carried out in triplicates and
physical examination. Subjects less than 18years had their mean zone diameter of inhibition was recorded.
consent sought from their relatives or guidance.
2.4.2. Determination of the Minimum
2.2. Methods Inhibitory Concentration (MIC)
The minimum inhibitory concentration of the extract was
2.2.1. Plant Processing and Extraction determined using the double fold dilution. An aliquot of 1mL
The stem G. mannii and T. avicennoides were collected of the reconstituted extract with 50% DMSO at a
washed and air-dried at room temperature at the Centre for concentration of 40 mg/mL was diluted serially to give
Genetic Engineering and Biotechnology, Federal University concentrations of 20, 10, 5, 2.5, 1.25, 0.625 and 0.3125
of technology, Minna, Niger State. The dried stem were then mg/mL in eight test tubes. One milliliter (1mL) of 18h
blended using blending machine to obtain a fine powder [16]. culture of bacteria previously adjusted to 0.5 McFarland
Fifty grams (50 g) of the plant powder was extracted with standard (1.0x106cfu/mL) was added to each of the test tubes
400mL of methanol and distilled water using reflux method and the content was mixed thoroughly. The tubes were
at a temperature of 45°C for 2 hours and the extract was incubated at 37°C for 4 h. The 9th test tube contained 1mL of
filtered using muslin cloth followed by further filtration using 50% DMSO with no extract served as negative control. The
whattman No 1 filter paper with pore size of 0.7µm to obtain 10th and 11th test tubes containing a solution of 5 mg/mL of
a fine filtrate. The filtrate was then concentrated using RE- Ampiclox and Amoxiline served as positive control. The
6000 rotary evaporator at 50°C and further concentrated procedure was repeated for the test and the control. The test
using the water bath at 45°C to ensure the extract is totally tube with the lowest concentration of the extract without
free from the solvent used in the extraction. visible turbidity of growth was taken as the MIC [19].
The resultant concentrates was kept in the refrigerator for
further use. 2.4.3. Determination of the Minimum
Bactericidal Concentration (MBC)
2.2.2. Qualitative Phytochemical Screening From each of the test tubes without any visible growth, a
of Extracts loopfull of the broth was aseptically inoculated on a sterile
Preliminary qualitative phytochemical screening which Mueller Hinton agar. The inoculated plates were incubated
involved performing simple chemical tests to detect the for 24hr at a temperature of 37°C. After incubation, the MBC
presence of secondary metabolites such as tannins, was determined as the lowest concentration with no visible
flavonoids, phenols, phenolic compounds, saponins, and growth on the plate [20].
glycosides, was carried out according to Trease and Evans
and Sofowora [10], [18].
2.5. Statistical Analysis
2.3. Quantitative Determination of
Phytochemicals Data obtained in this study were analysed using the IBM
Statistical Package for Social Science (SPSS) 20.0, 2011
Quantitative estimation of phytochemicals such as version (SPSS Inc., Chicago, Illinois, USA). Numerical data
alkaloids and saponins was carried out according to [21], were presented as mean±standard error of mean (SEM) of the
total phenolic content [23] and flavonoids using Aluminum triplicate.
Chloride colorimetric method [26].
11 Amuda Oladunni et al.: Phytochemical Screening and Antibacterial Activity of Extracts of Garcinia mannii and
Terminalia avicennoides on Some Oral Bacterial Pathogens

3. Results
Table 2. Percentage yield of the extracts of G. mannii and T. avicennoides.

Plants sample Solvent used Weight of extract (g) % yield

Methanol 5.32 10.64
G. mannii
Distilled water 5.23 10.46
Methanol 11.12 22.24
T. avicennoides
Distilled water 12.18 24.36

Table 3. Results of Phytochemical contents in aqueous and methanol extracts of G. mannii and T. avicennoides.

Plants Extract Alkaloids Phenols Tannins Saponins Cardiac glycosides

Methanol + + + + +
G. mannii
Aqueous + + + + +
Methanol + + + + +
T. avicennoides
Aqueous + + + + +

Table 3. Continued.

Plants Extract Phlobatannins Anthraquinones Steroids Terpenes flavonoids

Methanol + + + + +
G. mannii
Aqueous + + + + +
Methanol + + + + +
T. avicennoides
Aqueous + + + + +

Keys: +: Present, -: Absent

Table 4. Results of Quantitative phytochemical contents in methanol and aqueous extract of G. mannii and T. avicennoides (mg/100g).

Plants extract Total phenols Total flavonoids Alkaloids Tannins Saponins

TAA 706.945±147.58 255.333±17.61 151.606±14.07 169.550±2.91 4.807±0.02
TAM 788.364±1.80 233.111±12.27 125.482±1.90 155.283±5.67 7.012±0.90
GMA 900.518±8.90 198.352±2.13 120.312±3.15 138.500±6.03 8.280±0.28
GMM 1017.011±18.85 366.750±14.53 247.538±21.96 224.625±1.94 4.882±0.11

Values are expressed in mean±standard error of mean of duplicate determination.

Keys: TAA: Aqueous extract of T. avicennoides, TAM: Methanol extract of T. avicennoides, GMA: Aqueous extract of G. mannii, GMM: Methanol extract of
G. mannii.

Table 5. Mean zones of inhibition (mm) of methanol and aqueous extracts of Garcinia mannii against teste organisms.

GMA GMA GMA GMM GMM GMM Amoxicilin* Ampiclox*

Test organisms
20mg/mL 30mg/mL 40mg/mL 20mg/mL 30mg/mL 40mg/mL 5mg/mL 5mg/mL
M. luteus 13.67±0.33c 18.33±0.67d 21.00±1.53e 0.00±0.00a 0.00±0.00a 9.33±0.67b 26.33±0.33f 22.33±0.33ef
P. aeruginosa 11.00±1.00a 14.00±0.00bc 16.67±0.67d 11.33±0.67a 14.00±0.58bc 15.67±0.33c 13.67±0.33b 20.67±0.33e
S. mutans 12.33±0.33a 14.00±0.00b 22.33±0.33de 12.33±0.33a 21.33±0.67d 13.670.33c 25.67±0.33e 28.00±1.00f
K. pneumoniae 0.00±0.00a 0.00±0.00a 0.00±0.00a 19.67±0.67c 21.33±0.88cd 16.67±0.67b 19.00±1.00c 24.33±0.33d
S. pneumoniae 15.33±0.67b 16.33±0.33bc 11.33±2.03a 15.67±1.20b 21.00±0.58c 21.33±0.33c 15.670.33b 24.33±0.67d
S. pyogenes 15.33±0.88a 17.00±1.15b 16.00±1.15ab 17.33±0.88b 19.67±0.88c 19.67±0.67c 26.00±0.58d 27.67±0.33de

Values are expressed in mean±standard error of mean, values with the same superscript on the same row have no significant difference (p>0.05), n=3
GMA: Garcinia mannii aqueous extract, GMM: Garcinia mannii methanol extract
* Specification for Amoxicillin and Ampiclox are: ≤19mm (resistance) and ≥20mm (susceptible) (CLSI, 2012).

Table 6. Mean zones of inhibition (mm) of Methanol and Aqueous Extract of Terminalia avicennoides against test organisms.

TAA TAA TAA TAM TAM TAM Amoxicilin* Ampiclox*

Test organism
20mg/mL 30mg/mL 40mg/mL 20mg/mL 30mg/mL 40mg/mL 5mg/mL 5mg/mL
M. luteus 10.00±0.58a 15.33±0.67bc 16.67±0.33c 14.00±0.58b 17.67±0.88d 22.33±1.67e 26.33±0.33f 22.33±0.33e
P. aeruginosa 9.00±1.00a 14.67±0.67c 19.00±0.58d 20.67±1.20de 21.33±0.67e 19.00±0.58c 13.67±0.33b 20.67±0.33de
S. mutans 14.00±0.58a 16.00±0.58b 16.00±0.58b 22.00±1.15d 18.00±0.58c 19.33±0.33cd 25.67±0.33e 28.00±1.00f
K. pneumoniae 6.33±0.67a 12.67±0.33b 14.33±0.33c 23.67±0.33ef 24.33±0.33f 25.00±1.00g 19.00±1.00d 24.33±0.33f
S. pneumoniae 14.00±0.58a 18.33±0.33b 18.00±1.15b 21.33±0.67c 22.67±1.33cd 24.33±0.33d 15.670.33ab 24.33±0.67d
S. pyogenes 18.33±0.33a 21.33±0.88b 22.67±0.88bc 18.33±0.33a 21.33±0.88b 22.67±0.88bc 26.00±0.58c 27.67±0.33cd

Values are expressed in mean±standard error of mean, values with the same superscript on the same row have no significant difference (p>0.05), n=3
TAA: Terminalia avicennoides aqueous extract, TAM: Terminalia avicennoides methanol extract
* Specification for Amoxicillin and Ampiclox are: ≤19mm (resistance) and ≥20mm (susceptible) (CLSI, 2012).
 AASCIT Journal of Health 2017; 4(2): 8-16 12

Figure 1. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of Methanol extract of G. mannii (mg/mL).

Figure 2. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of aqueous extract of G. mannii (mg/mL).
13 Amuda Oladunni et al.: Phytochemical Screening and Antibacterial Activity of Extracts of Garcinia mannii and
Terminalia avicennoides on Some Oral Bacterial Pathogens

Figure 3. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of methanol extract of T. avicennoides (mg/mL).

Figure 4. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of aqueous extract of T. avicennoides (mg/mL).
 AASCIT Journal of Health 2017; 4(2): 8-16 14

4. Discussions force, electron flow, active transport, and coagulation of cell

contents [31].
The efficacy of a medicinal plant against any target disease The antibacterial potentials of these plants were judged
solely depends on the presence and concentration of the based on the zones of inhibition of bacterial growth of the
phytochemicals that are resident in that plant [21]. test organisms. The mean zones of inhibition of the isolates
Preliminary phytochemical screening of the extract reveals are a function of the relative antibacterial activities of the
the presence of alkaloids, phenols, tannins, cardiac extracts. The zones of inhibition is the area on the agar plate
glycosides, phlobatannins, anthraquinones, steroids, terpenes that remains free of bacterial growth. The size of the zones of
and flavonoids and saponins in all the extracts. (Table 3). The inhibition is usually related to the level of antibacterial
phytochemicals listed above except alkaloids have also been activity of the sample or extract; a larger zone of inhibition
reported in methanol extract of G. mannii [24]. Quantitative usually means the antibacterial potency of the extract is more
phytochemical analysis of the extracts shows high amount of [32].
total phenols and flavonoids, alkaloids tannins and saponins; The zones of inhibition of the extract ranged from
1017.01±18.85, 366.75±14.53, 247.54±21.96, 224.63±1.94 6.33±0.67 – 25.00±0.57 mm at a concentration of 20, 30 and
and 4.88±0.11mg/100g in methanol extract of G. mannii 40mg/ml in methanol and aqueous extract of G. mannii and T.
followed by aqueous extract of G. mannii, methanol and avicennoides (Table 5 and 6) which is an indication of the
aqueous extract of T. avicennoides with phenolic content of antibacterial potency of these extracts. The activity of these
900.52±8.90, 788.36±1.80 and 706.95±147.58mg/100g extract maybe attributed to the presence of the
respectively, alkaloids contents ranged from 120.31±3.15- phytochemicals at a relatively high concentration [34]. The
247.54±21.96mg/100g, tannins; 138.50±6.03-224.63±1.94 in antimicrobial activity of alkaloids which is also present in
four extract of the two plants while saponins have the lowest this extract have also been reported [35]. Mechanism of
concentration in all the extract ranging from 4.80±0.11- action of tannins involve the precipitation of protein to water
8.28±0.28mg/100g. soluble compounds and as a result, bacterial are inactivated
Phytoconstituents secreted by plants to protect them by the direct damage done to their cell membrane [36]. The
against pathogenic insects, bacteria, fungi or protozoa have Minimum Inhibitory Concentration (MIC) and Minimum
found applications in human medicine [25]. Some Bactericidal Concentration (MBC) which is the minimum
phytochemicals such as phenolic acids act essentially by concentration required to inhibit the growth of the
inhibiting adherence of organisms to the cells lining the microorganism or completely kill the microorganism
bladder, and the teeth, which ultimately lowers the incidence respectively was also recorded (Figure 1-4). The MIC and
of urinary-tract infections (UTI) and the usual dental caries. MBC of the methanol and aqueous extract of the plant ranged
It is worthy of note that antimicrobial activity results of the from 0.625-20mg/ml and 1.25-40mg/ml respectively. The
same plant part tested most of the time varied from lower the MIC and MBC the higher the potency of the
researcher to researcher. This is possible because extract. This MIC and MBC values also indicate that the
concentration of plant constituents of the same plant organ extract of this plant contains bioactive compounds which can
can vary from one geographical location to another and from serve as a precursor in the synthesis of new drugs [24].
one part of the plant to another depending on the age of the Therefore, the presence of these phytochemicals in good
plant, differences in topographical factors, the nutrient quantity is an indication of its medicinal potency and why it
concentrations of the soil, extraction method as well as is used by traditional herbalist in treating many ailments [19].
method used for antimicrobial study [27].
Different mechanisms of action of phytochemicals have 4.1. Conclusions
been suggested. They may inhibit microorganisms, interfere
with some metabolic processes or may modulate gene From the results of this study, aqueous and methanol
expression and signal transduction pathways [28]; [29]; and extracts of T. avicennoides and G. mannii possesses active
[30]. Phytochemicals may either be used as chemotherapeutic phytochemical constituents and antibacterial activities
or chemo preventive agents with chemoprevention referring against the selected oral bacterial pathogens, it can therefore
to the use of agents to inhibit, reverse, or retard be concluded that methanol and aqueous extracts of these
tumorigenesis. Plant extracts and essential oils may exact plants may be used as chewing sticks since it contains
different modes of action against bacterial strains, such as bioactive components that can also inhibit the growth and
interference with the phospholipids bilayer of the cell activities of the microorganisms in the oral cavity. Also, the
membrane which has as a consequence may increase cell plants can also be used in the manufacture of herbal paste
permeability and exudation of cellular constituents, damage and drugs which can be use in the treatment of ailments
or changes in the enzymes involved in the production of caused by these microorganisms.
cellular energy and synthesis of structural components, and 4.2. Conflict of Interest
destruction or inactivation of genetic material. In general, the
mechanism of action is considered to be the disturbance of The author has declared there is no conflict of interest.
the cytoplasmic membrane, disrupting the proton motive
15 Amuda Oladunni et al.: Phytochemical Screening and Antibacterial Activity of Extracts of Garcinia mannii and
Terminalia avicennoides on Some Oral Bacterial Pathogens

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