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he increasing use of chemical drugs causing bacterial resistance to the drugs caused side effects
Vol. 26 - No. 4
1ZabolMedicinalPlantsResearchCenter
ZabolUniversityofMedicalSciences, Zabol, Iran
2DepartmentofPharmacognosy, FacultyofPharmacy
ZabolUniversityofMedicalSciences, Zabol, Iran
3Pediatric Infections Research Center
Mofid Children Hospital, Shahid Beheshti
University of Medical Sciences, Tehran, Iran
4DepartmentofRangeandWatershedManagement
FacultyofNaturalResources, UniversityofZabol, Iran
5Zabol University of Medical Sciences, Zabol, Iran
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The Prosopis farcta were collected during the mature period, May 2013, from the area surrounding
Hamun Lake, Zabol (Coordinates: 31 1 43 N, 61
30 4 E), in Sistan and Baluchestan Province of Iran
(Figure 1). The plant was taxonomically identified
by the Ferdowsi University Mashhad Herbarium,
Iran (Voucher No: 25685). Twenty grams of each
parts viz. roots, leaf, pods and seeds were powdered
separately and then dissolved in 200 mL methanol
85% using a shaker water bath for 24 hours at 25
C. After filtration with Whatman No. 1 filter paper,
the resulting solutions were concentrated by a rotary
evaporator at 40 C for 35 min to remove solvents
from the extracts. Solid extracts (residues of plant
extracts) were dissolved in 20 mL of distilled water.
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Four hundred and seventy-seven clinical specimens such as burn, wound, urine, pus and throat
swab were collected from patients attended in emergency Hospital and Internal Laboratory of Hospital
and Central Laboratory in Zabol city (Iran) for different bacteriological examinations (Table I). Standard
isolation protocols were used for all the samples.
Identification of S. aureus was approved by standard techniques accord on colonial and microscopic
morphology, coagulase test and biochemical activities.17 Detection of MRSA was done by the use of
Cefoxitin disc (30 g) diffusion test. The strains of
S. aureus with a zone diameter of <19 mm based
on Clinical and Laboratory Standard Institute (CLSI)
were taken to show methicillin-resistance.19 All
strains of S. aureus were confirmed as methicillin
resistant by oxacillin agar dilution using Muller Hinton agar (beef extract, acid hydrolysate of casein
17.5g/L, starch 1.5 g/L, agar 17.0 g/L, final 7.30.1
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Table I.The number and percentage of isolated MRSA from various clinical specimens.
S. aureus
53
38
11
4
1
107
50.69
84.44
4.33
28.57
1.66
22.43
9
4
4
0
1
18
16.98
10.52
36.36
0.00
100
16.82
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Antimicrobial activity was based on the disc diffusion method 20 using a cell suspension of microorganisms. The concentration of the cell suspension
was equilibrated to a 0.5 McFarland standard and 50
L of each microorganisms suspension was spread
on a Mueller-Hinton agar plate. In addition, 50 L of
diluted methanolic extracts was pipetted onto sterile
paper discs (6 mm in diameter), which were dried
in an open sterile Petri dish in a biological laminar
flow bench. Discs were placed on the surface of
inoculated plates and incubated at 37 C for 24 h.
Diameters (mm) of the zones of bacterial inhibition
minus the discs diameter were recorded.21
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at 25 C) supplemented with 2% NaCl. The concentrations tested ranged from 1 g/mL to 16 g/mL
of oxacillin. The strains having minimum inhibitory
concentration (MIC) 4 g/mL were taken as resistant. Among 477 clinical specimens, of the 107 specimens (22.43%) were S. aureus that from these 18
specimens (16.82%) were MRSA.
N.
104
45
254
14
60
477
Burn
Wound
Urine
Pus
Throat
Total
MRSA
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The MIC values for each plant extracts were determined by microbroth method. Residues of plant extracts were dissolved in 20 mL of distilled water. All
extracts are tested at 1000 g/mL 22 and serially diluted two-fold to 1.95 g/mL in a 96-multiwell polystyrene flat-bottomed microplate (Sigma-Aldrich, St.
Louis, MO, USA) after which 100 L (1106 CFU/mL)
of bacteria are added to each of them. Preincubation
absorbance values were read from an ELISA reader.
The microplates were then incubated overnight at
37 C and absorbance values were read after 24 h.
The MIC values are recorded as the lowest concentration of the extract that completely inhibited bac-
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Number of specimens
Type of specimens
Results
Preliminary phytochemical analysis
The results of preliminary phytochemical analysis
of different parts (roots, leaf, pods and seeds) of the
P. farcta showed that roots methanolic extract demonstrated high concentrations of flavonoids, saponins,
phenols and moderate concentrations of alkaloids,
tannins, glycosides and resins. The leaf and pods
methanolic extract showed a high concentration of
saponins and flavonoids and moderate concentrations
of alkaloids, resins, glycosides, tannins and phenols.
Seeds methanolic extract presented high concentrations of alkaloids, tannins, glycosides and moderate
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Table II.Results of phytochemical analysis of extracts from root, leaf, pod and seed of the P. farcta.
Methanolic extracts
Chemical Compounds
Saponins
Tannins
Flavanoids
Glycoside
Alkaloids
Phenols
Resin
Root
Leaf
Pod
Seed
++
+
++
+
+
++
+
++
+
++
+
+
+
+
++
+
++
+
+
+
+
+
++
+
++
++
+
+
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Figure 2.Inhibition zones of all parts of the Prosopis farcta. A) roots; B) leaf; C) pods; D) seeds.
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In this study, all isolated MRSA were entirely sensitive to vancomycin. Similar results were achieved
for vancomycin assay in previous studies.23, 24 Vancomycin has been the most effective therapeutic
agent against MRSA. However, increased use of vancomycin has set a basis of selection for vancomycinresistance in MRSA.
S. aureus is a versatile human pathogen; it was
powerfully considered as a major reason of nosocomial infection. In recent years, the prevalence of
MRSA has enhanced universal as it is evident from
numerous supervision studies.25-33 However, infection with MRSA changes greatly from one geographic location to another, from hospital to hospital.34
Today, some antimicrobial agent such as vancomycin and teicoplanin are still exclusively efficient
against MRSA; thus, this pathogen can cause critical
infection in different body systems in patients. The
emergences of expanding in antibiotic resistance
due to many researchers were examined optional
accesses to treat staphylococcal infection. Plant extracts have been extensively studied as natural products.
Zuo et al.35 performed a study on potential antibacterial activity of aerial parts of the Chinese medicinal plants against clinical isolates of MRSA. They
reported that the MIC for Dendrobenthamia capitata, Elsholtzia rugulosa, Elsholtzia blanda, Gera-
Discussion
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References
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Conclusions
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resistant Staphylococcus aureus and multi-drug resistant Pseudomonas aeruginosa. Asian Pac J Trop Biomed 2013;3:663-7.
38. Rad JS, Alfatemi SMH, Rad MS, Iriti M. In-vitro antioxidant and
antibacterial activities of Xanthium strumarium L. extracts on
methicillin-susceptible and methicillin-resistant Staphylococcus
aureus. Ancient Sci Life 2013;33:107-11.
39. Darogha SN. Phytochemical analysis and antibacterial activity of
some medicinal plants against methicillin-resistant Staphylococcus aureus. Tikrit J Pharma Sci 2009;5:116-26.
40. Al-Ameri AK. Evaluation of antimicrobial activity of aqueous extract of Prosopis farcta pods. Tikrit J Pharma Sci 2006;2:78-84.
41. Sharifi-Rad J, Hoseini-Alfatemi SM, Sharifi-Rad M, Teixeira da
Silva JA. Antibacterial, antioxidant, antifungal and anti-inflammatory activity of crude extract from Nitraria Shoberi fruits. 3
Biotech 2014 [Epub ahead of print].
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