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Advances in Environmental Biology, 5(2): 491-495, 2011
ISSN 1995-0756
This is a refereed journal and all articles are professionally screened and reviewed ORIGINAL ARTICLE
Antibacterial Activity of Extracts of Ajuga Iva, and Teucrium Polium
1
ZERROUG M.M., 2ZOUAGHI M., 2BOUMERFEG S,2BAGHIANI A, 3NICKLIN J and
2
ARRAR L
1
Laboratory of Applied Microbiology, Department of Microbiology, University Ferhat Abbes of Setif,
Setif, ALGERIA
2
Laboratory of Applied Biochemistry, Department of Biochemistry, University Ferhat Abbes of Setif, Setif,
ALGERIA
3
School of Biological and Chemical Sciences, Birkbeck College, University of London UK.
ZERROUG M.M., ZOUAGHI M., BOUMERFEG S, BAGHIANI A, NICKLIN J and ARRAR
L: Antibacterial Activity of Extracts of Ajuga Iva, and Teucrium Polium
ABSTRACT
Antibiotics provide the main basis for the therapy of microbial (bacterial and fungal) infections.
Since the discovery of these antibiotics and their use as chemotherapeutic agents there was a belief
in the medical fraternity that this would lead to the eventual eradication of infectious diseases.
However, overuse of antibiotics has become the major factor in the emergence and dissemination of
multi-drug resistant strains of several groups of microorganisms. Many plants which are used in
traditional medicine contain antimicrobial compounds. In this study, the antibacterial activity of
methanolic extracts of Teucrium polium and Ajuga iva were tested against five bacteria, E. coli MC
4100, Bacillus subtilis, Pseudomonas diminutus, Paracoccus paratrophus and Micrococcus luteus. T.
polium and A. iva plants were collected and allowed to dry in the dark at room temperature. Dried
plant material (100g) was added to 1 L of methanol and incubated at room temperature for three days.
The crude solution was filtered through muslin cloth, and the filtrate evaporated to dryness. The dried
material was dissolved in 2 ml of methanol. Bacterial suspensions (100 µl) were spread on tryptone
soya agar (TSA) medium. Plant extracts (10 µl) were applied to discs of filter paper and placed on
agar plates containing the microorganisms. The plates were incubated at 37°C for 48h. After incubation
the zones of inhibition around the discs was measured. Extracts of T. polium gave zones of inhibition
against Bacillus subtilis, Micrococcus luteus and Paracoccus paratrophus of 3.7, 2.0 and 2.0 mm,
respectively. A. iva extract only inhibited the growth of Paracoccus paratrophus, giving a zone of
inhibition of 3.0.mm. The present results showed that extracts of T. polium inhibited the growth of
three bacterial species. Extracts of A. iva, on the other hand, inhibited only one bacterium.
Key word: Ajuga iva, antibacterial activity, plant extracts, Teucrium polium.
Introduction would lead to the eventual eradication of
infectious diseases. However, overuse of antibiotics
Antibiotics provide the main basis for the has become the major factor in the emergence
therapy of microbial (bacterial and fungal) and dissemination of multi-drug resistant strains of
infections. Since the discovery of these antibiotics several groups of microorganisms [10,13].
and their use as chemotherapeutic agents there Numerous studies have shown that aromatic and
was a belief in the medical fraternity that this medicinal plants are sources of diverse nutrient
Corresponding Author:
ZERROUG M.M., Laboratory of Applied Microbiology, Department of Microbiology,
University Ferhat Abbes of Setif, Setif, ALGERIA
Email: med.zerroug@gmail.com
Adv. Environ. Biol., 5(2): 491-495, 2011 492
and non nutrient molecules, many of which Chemical Sciences, Birkbeck College University of
display antioxidant and antimicrobial properties London, UK
which can protect the human body against both
cellular oxidation reactions and pathogens. Plants Antibacterial Assay:
are rich in a wide variety of secondary
metabolites such as tannins, alkaloids and The dried material was dissolved in 2 ml of
flavonoids, which have been found in vitro to methanol and was tested against the five
have antimicrobial properties [2,26,21]. Compounds mentioned bacteria. Bacterial suspensions (100 µl)
that inhibit pathogens but have little toxicity to were spread on Tryptone Soya Agar (TSA)
host cells are candidates for developing new medium. Plant extracts (10 µl) were applied to
antimicrobial drugs [21]. There are several reports discs of filter paper (Whatmann AA, 6 mm
on the antimicrobial activity of different herbal diam.) and placed on agar plates containing the
extracts [2]. A number of phytotherapy manuals microorganisms. The plates were incubated at
have mentioned various medicinal plants for 37°C for 48h.
treating infectious diseases owing to their
availability, few side effects and low toxicity Results and discussion
[15].
Thus, it is important to characterize different After incubation the zones of inhibition
types of medicinal plants for their antioxidant and around the discs was measured. Extracts of T.
antimicrobial potential [2,25,21]. In the light of polium gave zones of inhibition against Bacillus
evidence for the rapid global spread of isolates subtilis, Micrococcus luteus and Paracoccus
of pathogenic microorganisms with antibiotic paratrophus of 3.7, 2.0 and 2.0 mm, respectively.
resistance, the need to find new antimicrobial A. iva only inhibited the growth of Paracoccus
agents is of paramount importance. However, the paratrophus, giving a zone of inhibition of
past record of rapid, widespread emergence of 3.0.mm (Figure 1, Table 1).
resistance to newly introduced antimicrobial agents The antimicrobial compounds from plants may
indicates that even new families of antimicrobial inhibit bacterial growth by different mechanisms
agents will have a short life expectancy [6]. For than those presently used. Antimicrobials therefore,
this reason, researchers are increasingly turning may have a significant clinical value in treatment
their attention to herbal products, looking for new of resistant microbial strains. In particular, the
leads to develop better drugs against strains of antimicrobial activity of plant oils and extracts
microorganisms with multi drug resistance [4]. have formed the basis of many applications
According to the world health organization including raw and processed food preservation,
(WHO), medicinal plants would be the best pharmaceuticals, alternative medicine, and natural
source for obtaining a variety of drugs [19]. therapies [20].
Therefore, in this report information on the T. polium L. (Lamiaceae) has numerous
antimicrobial activity of methanolic extracts of pharmacological properties. These include calcium
two plants is given. antagonistic, anorexic, intestinal motility and
hypertension, anti-ulcer, anti-inflammatory,
Materials and methods antipyretic and antibacterial actions, glycemic and
hypolipidemic effects; treat liver disease, and
Plant Material: diabetes. The extract is also used as an
antiemetic, an antispasmodic, an anti-inflammatory,
Teucrium polium and Ajuga iva plants were an antipyretic, an analgesic, and an anti-
collected and allowed to dry in the dark at room carminative [17].
temperature. Dried plant material (100g) was The antibacterial activity of T. polium extracts
added to 1 litre of methanol and incubated at can be attributed to its contents in flavonoids. In
room temperature for three days. The crude general the methanol extraction of T. polium
solution was filtered through muslin cloth, and plant material yielded to flavonoids [22]. The
the filtrate evaporated to dryness. obtained results were similar to the results in
other studies, the methanolic extract of T. polium
Bacteria Strains: inhibited the growth of several bacteria with
different minimal inhibitory concentration (MIC)
Five bacterial strains were used in this study, This extracts inhibited the growth of
E. coli MC 4100, Bacillus subtilis, Pseudomonas Staphylococcus aureus, Salmonella typhi with a
diminutus, Paracoccus paratrophus and MIC of 40 mg/mL, this concentration was 10
Micrococcus luteus obtained from the laboratory mg/mL Bordetella bronchiseptica ,and Bacillus
of Dr. Jane Nicklin, School of Biological and anthracis. This cocnentration of 10 mg/mL
Adv. Environ. Biol., 5(2): 491-495, 2011 493
represents also the minimal bactericidal naringenin-7-O-_-D-glucopyranoside and nicotiflorin)
concentration (MBC) against Bacillus anthracis. was tested against several strains of K.
The plant extract was also active against pneumoniae, all these flavonoids, showed in vitro
Bordetella bronchiseptica. The hydroalcoholic antimicrobial activity similar to that produced by
extract of T. polium had a relatively satisfactory the control antibacterial (ofloxacin) at the
effect on Salmonella typhi. [7]. All flavonoids concentration of 32-64 μg/ml [16]. Similarly, the
isolated from leaves of Psidium guajava inhibited aqueous extract of T. polium inhibited the growth
with bacteriostatic mode of action all of the fish of Saccharomyces cerevisiae (Ki = 29 µg/l) and
pathogens used in study, Aeromonas hydrophila; Yarrovia lipolytica (Ki =61 µg/l) [10], in contrast
Aeromonas salmonicida subsp. salmonicida ATCC the ethanolic extraxt of T. polium had no effect
14174; Flavobacterium columnare ATCC 23463; on Micrococcus luteus NRRL B- 4375 [1].
Lactococcus garvieae ATCC 49156; Streptococcus Ethnopharmacological surveys have revealed
agalactiae; Vibrio salmonicida ATCC 43839 [18]. that some 20 species of Ajuga plants are used in
Maximum activity of flavonoid fraction of traditional medicine mostly in Africa, Asia and
callus tissue from Gossypium species was China). In North Africa, Ajuga plants are used to
observed of several tested bacteria, Bacillus cerus treat diabetes and hypertension [9,24]. Other
(NCIM 2156), Staphylococcus aureus (NCIM reported activities of Ajuga plants include
2654) Staphylococcus epidermidis (NCIM 2493), antibacterial, antifungal, anti-inflammatory,
Mycobacterium smegmatis (NCIM 5138), antimalarial/antiplasmodial, antimycobacterial,
Pseudomonas aeruginusa (NCIM 5032), Proteus antioxidant, antipyretic, larvae and insect
vulgaris (NCIM 2027), Salmonella typhimurium antifeedant and insect growth inhibitor activity
(NCIM 2501), Escherichia coli (NCIM 2027) [5]. [11].
In the same way the propolis products which This activities are related to there contents in
contained flavonoids, expressed bactericidal activity active compounds, they contains a wide range of
against B. subtilis and S. aureus with a zone of compound such as ajugapyrin A, bracteonin A
inhibition of 19 mm and 17 mm respectively and lupulin C and Iridoids which had a wide
[14]. In other study six flavonoids (7- range of biological and pharmacological activity.
dimethoxyflavanone-4’-O-_-D-glucopyranoside, 5,7- The plant contains also, a class of secondary
dimethoxyflavanone-4’-O-[2’’-O-(5’’’-O-trans- metabolites which are produced by plants
cinnamoyl)-_-D-apiofuranosyl]-_-Dglucopyranoside, primarily as a defence against herbivores or
5,7,3’-trihydroxyflavanone-4’-O-_-D-glucopyranoside, against infection by microorganisms [23,11].
Fig. 1: Antibacterial effect of Teucrium polium (8) and Ajuga iva (8) against Bacillus subtilis (2),
Micrococcus luteus (5) and Paracoccus paratrophus (6).
Adv. Environ. Biol., 5(2): 491-495, 2011 494
Table 1: Antibacterial effect of T. polium and A. iva extracts
Bacterial strains Teucrium polium extracts Ajuga iva extracts
Bacillus subtilis 3.7 0.0
E. coli MC 4100 0.0 0.0
Micrococcus luteus 2.0 0.0
Paracoccus paratrophus 2.0 3.0
Pseudomonas diminutus 0.0 0.0
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