According to World Health Organization, (Santose, et. al., 1995) medicinal plants would be the best source
to obtain a variety of drugs. About 80% of individuals from developed countries use traditional medicines,
which have compounds derived from medicinal plants. Therefore, such plants should be investigated to
better understand their properties, safety and efficiency (Elof, et al., 1998).
Klein (1998) has taken view and supported it by strong experimental evidence, that synergism occurs only
when the microorganism tend to become drug-fast and is related to inhibition by the second of resistance
microorganism surviving the action of first. Secondly, when drug-fastness does not develop with some
facility, synergism commonly does not occur and some degree of antagonism may be observed. The
consequence of combining drug is dependent upon the strains of micro-organisms upon the condition under
which the organism is subjected to the action of drug.
Salsola kali (Family: Chenopodiaceae) is an annual herb commonly found in Bahawalpur (Cholistan
desert) South Punjab, Pakistan. This plant has medicinal importance and is used in traditional medicinal
system of their native region by various hakims to fight against disease. This plant is used to investigate the
antibacterial activity because of its medicinal properties.
Nascimento, G.F, et al., (2000) evaluated the antimicrobial activity of plant extracts and phytochemicals
with antibiotic susceptible and resistant microorganisms. The possible synergistic effects when associated
with antibiotics were studied with Achillea millifolium (yarrow), Melissa officinalis (lemon-balm), Ocium
basilucum (basil), Psidium guajava (guava), Punica granatum (pomegranate), Rosmarinus officinalis
(rosemary), Salvia officinalis (sage), Syzgyum joabolanum (jambolan) and Thymus vulgaris (thyme). The
highest antimicrobial potentials were observed for the extracts of Caryophyllus aromaticus and Syzygyum
joabolanum, which inhibited 64.2 and 57.1% of the tested microorganisms, respectively, with higher
activity against antibiotic resistance bacteria (83.3%).Association of antibiotics and plant extracts showed
synergistic antibacterial activity against antibiotic resistance bacteria. Pseudomonas aeruginosa was
inhibited by clove, jambolan, pomegranate and thyme extracts. This inhibition was observed with the
individual extracts and when they were used in lower concentrations with ineffective antibiotics.
Soberon J.R. et al., (2007) determined the antibacterial and cytotoxic activities of aqueous and ethanolic
extracts of northwestern Argentinian plants used in folk medicine and compared with different commercial
antibiotics. Tripodanthus acutifolius aqueous extract has lower inhibitory concentrations than cefotaxim
against Acinetobacterfreundii. Tripodanthus acutifolius tincture showed lower MIC and minimal
bactericidal concentration (MBC) than cefotaxim for Pseudomonas aeruginosa. This extract also showed a
MIC/MBC lower than oxacillin for Staphylococcus aureus. The cyto-toxicity of all extracts was compared
with that of commercial antibiotics. Rutin (3,3',4',5,7-pentahydroxy flavone 3-beta-rhamnosilglucoside ,isoquercitrin
(3,3',4',5,7-pentahydroxyflavone 3-beta-glucoside) and a terpene would be partially responsiblefor the antibacterial activity of T. acutifolius infusion.
Bonjar, et al., (2004) reported that forty-five species of 29 plant families used in the traditional medicine
by Iranian people, showed antibacterial activities against one or more of the bacterial species: Bacillus
cereus, Bacillus pumilus, Bordetella bronchiseptica, Escherichia coli, Klebsiella pneumoniae, Micrococcus
luteus, Pseudomonas aeruginosa, Pseudomonas fluorescens, Serratia marcescens, Staphylococcus aureus
and Staphylococcus epidermidis. No plant showed activity against Serratia marcescens, Bordetella
bronchiseptica being the most susceptible species. All extracts showed the same activity 18 months later.
Camporese, et al., (2003) reported that twenty-one extracts from seven herbal drugs, Aristolochia trilobata
(Aristolochiaceae) leaves and bark, Bursera simaruba (Burseraceae) bark, Guazuma ulmifolia
(Sterculiaceae) bark, Hamelia patens (Rubiaceae) leaves and Syngonium podophyllum (Araceae) leaves and
bark, used in traditional medicine of Belize (Central America) as deep and superficial wound healers, were
evaluated for their anti-bacterial properties. Activity was tested against standard strains of Escherichia coli
ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923 and
Enterococcus faecalis ATCC 29212. Almost all the extracts were able to inhibit the growth of one or more
of the bacterial strains, except that of Enterococcus faecalis.
MATERIAL AND METHODS
Microbial strains of Staphylococcus aureus (ATCC 25923) (S.aureus). Escherichia coli (ATCC 2592) (E.
coli), pseudomonas aeruginosa (ATCC 27853) (P. aeruginosa), Streptococcus pneumoniae (ATCC 49619)
(S. pneumoniae) Bacillus subtilus (ATCC 6051) (B. subtilus) and Sarcina lutae ATCC 9341 (S. lutae) and
Streptococcus mutans (ATCC 020572) were obtained from Pediatrics, Microbiology Laboratory, Mayo
Fresh plants of Salsola kali (whole plant (PM # 086) was collected from South Punjab (Bahawalpur Road )
on 6th June 2005 and Senecio chrysanthemoides (whole plant, PM # 185) and Galium asperuloides (PM #
0234) were collected from area between Nathia Gali and Khanuspure , Muree Hill , Pakistan on 2nd
October 2005. The plants were identified by Mir Ajab Ali Khan, Professor of Botany Quaid-e-Azam
University, Islamabad, Pakistan, Dr. Zaheer-ud-din , Professor of Botany Government College Lahore,
Pakistan and voucher specimen were deposit in Prem Madan Herbarium of Lahore College for Women
University, Lahore Pakistan.
Salsola kali (whole Plant), Senecio chrysanthemoides (whole Plant) , Galium asperuloides (whole plant)
and Heliotropium strigosum (whole Plant) were air dried, finely gained extracted methanol by soxhelt
extraction to yield 15% ,20%, 18% and 25% solvent free extract.
Anti bacterial activity was determined by agar well diffusion method [Norsel and Messley .1977]. This test
was performed in triplicate by spreading 12-18 hour old pathogenic bacterial cutters containing
approximately 106 - 1010 colony forming unit (CFU/ml) on the surface of nutrient agar plates well (4mm)
were dug in the media with the help of sterile metallic borer.
Test samples of different concentrations prepared in Methanol were added (50µl) in their respective wells
pure methanol was used as negative control (3mm) other wells were supplemented with reference
compounds i.e. Ampicillin, Amoxicillin, Levofloxin, Tetracycline, Vancomycin, Ciprofloxacin and
Penicillin as positive control.
Synergistic Activity against bacterial strains was determined by taking equal amount (50µl (1:1))of plant
extracts by agar well diffusion method as described before.
RESULTS AND DISCUSSIONS
Seven pathogenic bacterial strains (Staphylococcus aureus, Escherichia. coli, Bacillus subtilis,
Streptococcus pneumoniae , Sarcina lutae, Streptoccus mutans and Pseudomonas aeruginosa) were used
in this study and methanolic extract of Salsola kali with three combinations of methanolic extracts of
other plants (Salsola kali + Galium asperuloides, Salsola kali + Senecio chrysanthemoides and Salsola kali
+ Heliotropium strigosum with concentrations (250, 100, 50, 10, 5, 1.0, 0.5 µg/ml) were used against each
of the seven bacterial strains.
The crude methanolic extract of Salsola kali showed highest activity against S.mutans, S.aureus, B.subtilis
and S. pneumoniae while it showed moderate bactericidal activity against P.aeruginosa. The crude
methanolic extract of Salsola kali inhibits the growth of S.lutae and E. coli. (Table-01&05)
Table – 01 Zone of inhibitions of crude methanol extracts of Salsola kali
The crude methanolic extract of Senecio chrysanthemoides showed highest antibacterial activity against
S.aureus, S.mutans and S.pneumoniae . The plant extract inhibits the growth of P.aeruginosa, E.coli
,B.subtilus and S.lutae (Table -02&05)
Table -02 Zone of inhibition of crude methanol extracts of Senecio chrysanthemoides
The methanolic extract of Galium asperuliodes showed moderate activity against S.aureus, S. pneumoniae,
S.lutae and E.coli . While this extract showed good activity by increase the concentration of the extract
with P.aeruginosa , S.mutans and B.subtilis. (Table-04 &05)
Table -04 Zone of inhibition of crude methanol extracts of Galium asperuliodes
Synergistic activity: The crude methanolic extract of Salsola kali and Heliotropium strigosum showed
good activity against S.lutae, S.pnumiae, E.coli , B.subtilus and S.mutans (0.5µg/ml). The crude
methanolic extract of Salsola kali with Heliotropium strigosum showed moderate activity against S.aureus
and P.aeruginosa . (Table-06& 09)
Table -06 Zone of inhibition of crude methanol extracts of Salsola kali and Heliotropium strigosum
The crude methanolic extract of Salsola kali and Senecio chrysanthemoides showed highest activity
against E.coli, P.aeruginosa B.subtilus, S.aureus and S.pneumoniae but inhibit the growth of S.lutae and
S.mutans (Table- 07 & 09)
The methanolic extract of Salsola kali and Galium asperuloides showed highest activity against
P.aeruginosa E.coli, Sarcina lutae, Streptococcus mutans, Streptococcus pneumoniae and B.subtilis . The
Plant MIC value ranges from 0.5µg/µl-1.0µµg/l. While this extract showed moderate activity against
S.aerus .( Table 08 & 09)
Table -08 MIC value of crude methanol extracts of Salsola kali and Galium asperuloides.
The MIC values of test medicinal plants showed best activity as compared with the different discs of the
antibiotics (Ampicillin, Amoxicillin, Levofloxacin, Tetracycline, Vancomycin, Ciprofloxacin and
Penicillin) against the test strains of bacteria. (Table -10)
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