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International Journal of Advanced Research in Botany (IJARB)
Volume 1, Issue 1, Jul-Sep 2015, PP 10-14
www.arcjournals.org
©ARC Page | 10
Inhibitory Effects of Echinacea Angustifolia Essential Oils on the Growth
of Five Pathogenic Organisms: Coliform Spp, Pseudomonas Spp,
Saccharomyces Cerevisiae, Zygosaccharomyces Bailii and Lactobacillus
Plantarum
Bachir Raho G1, Benattouche Z1, Bevilacqua A2, Corbo MR2, Sinigaglia M2,
Pignatiello S2
1Department of Biology, University of Mascara, Algeria.
2Department of Food Science, Faculty of Agricultural Science, Foggia University, Italy.
bachir_raho@yahoo.fr
Abstract: Echinacea commonly called the Purple coneflowers, is a genus of nine species of herbaceous plants
in the Family Asteraceae. Three of them are important in commerce, with the majority of wild harvest being E.
angustifolia. It has been used for a variety of ailments, including toothache, coughs, colds, sore throats,
snakebite, and as a painkiller. In the current study, in vitro inhibitory activity of Echinacea angustifolia
essential oils were screened against Coliform spp, Pseudomonas spp, Saccharomyces cerevisiae (EC1118),
Zygosaccharomyces bailii (DSM 70492) and Lactobacillus plantarum (DSM2601). Agar well diffusion assay
was adopted for the study. E. angustifolia oils showed very weak antimicrobial activity against the
microorganisms tested with diameter of inhibition zone not exceeding 3 mm. The highest activities were
observed for Z. baillii and S. cereviceae at a concentration of 10 and 100 ppm respectively, while for the rest of
the strains the diameter of inhibition zone were ranged 1 and 2.5 mm, except Coliform spp which was not
affected by the presence of the essential oil at a concentration of 50 ppm. The low bacteriostatic effect of this
plant essential oil against some of the most important causes of infections provides an exciting potential for the
future, especially in the light of the shift away from commonly used antibiotics and the move towards more
natural alternatives.
Keywords: Echinacea angustifolia oils; antimicrobial activity; organisms.
1. INTRODUCTION
In the last three decades, although pharmacological industries have produced number of new-
antibiotics, but microbial resistance to these drugs by microorganisms has increased because of
genetic ability of the bacteria to acquire and transmit the resistance against to drugs, which are utilized
as therapeutic agents (Nascimento et al ., 2000; Tajehmiri et al ., 2014). Herbal treatment is one
possible way to treat diseases caused by multidrug resistant bacteria (Olukoya et al., 1993). Medicinal
plants play a key role in the human health care. About 80% of the world population relies on the use
of traditional medicine, which is predominantly based on plant material (Kumar, 2014). Echinacea
belongs to the Asteraceae, a family important to commerce for its many medicinal and culinary herbs.
There are nine species of Echinacea known this time, but only three are marked in the medicinal herb
trade. E. angustifolia, E. purpurea and Echinacea pallida (Miller., 2000). Echinacea has a long
history of medicinal use for a wide variety of conditions, mainly infections, such as syphilis and septic
wounds, but also as an ‘‘anti-toxin’’ for snakebites and blood poisoning. Traditionally, Echinacea was
described as an ‘‘anti-infective’’ agent, and was indicated in bacterial and viral infections, but the
current interest in the medicinal use of Echinacea is focused on its immunostimulant (increasingly
described as immunomodulatory) effects, particularly in the treatment and prevention of the common
cold, influenza and other upper respiratory tract infections (Barnes et al ., 2005). The chemistry of
Echinacea species is well known and caffeic acid derivatives, flavonoids, polyacetylenes, alkamides,
pyrrolizidine alkaloids, polysaccharides and glycoproteins were isolated and characterized
(Lucchesini et al ., 2009).
Therefore, this study was conducted to evaluate the antimicrobial activity of Echinacea angustifolia
extracts against Coliform spp, Pseudomonas spp, Saccharomyces cerevisiae,
Zygosaccharomyces bailii and Lactobacillus plantarum.
Bachir Raho G et al.
International Journal of Advanced Research in Botany (IJARB) Page | 11
2. MATERIALS AND METHODS
Essential oils
The essential oils (EO) of Echinacea angustifolia were provided by a commercial company,
Farmalabor (Canosa di Puglia, Italy) as liquid extract.
Tested microorganisms
To assess the antimicrobial properties of E. angustifolia EO, three strains of bacteria and two yeasts
were used in the study: Coliform spp and Pseudomonas spp isolated by the Laboratory of Applied
Microbiology (University of Foggia, Italy), while Saccharomyces cerevisiae EC1118 (Lallemand
Inc.), Zygosaccharomyces bailii DSM 70492 and Lactobacillus plantarum DSM2601 were procured
from the German Collection of Microorganisms and Cell Cultures (Deutsche SammLung von
Mikroorganismen und Zellkulturen GmbH, DSMZ, Germany). Microbiological media were
purchased from Oxoid Ltd (Basingstoke, UK) and Biolife (Milan, Italy).
Antimicrobial Activity Assay
The antimicrobial activity of the E. angustifolia EO was determined with the agar-well diffusion
method. Cultures of microbes age 24 h were inoculated separately on the solidified Nutrient agar
(except L. plantarumin MRS) on each Petri dish by streaking using sterilized cotton swabs. Two wells
were made in the solidified agar using a sterile borer and each hole was filled with 10, 50 or 100 ppm
of plant extract. The control was set in parallel without essential oil. The plates were then incubated at
37°C for the bacteria and 25°C for the yeast, for 24 h. The sensitivity of the test microbes to the
extracts were determined by measuring the diameters of the zone of inhibition surrounding the wells
in millimeter (mm).
3. RESULTS AND DISCUSSION
The in vitro antimicrobial activity carried out by agar-well diffusion method of the essential oil
resulted in a range of growth inhibition pattern against tested microorganisms summarized in Table
below.
Table. Results of agar-well diffusion test of various concentration of E. angustifolia essential oil against
bacteria and yeasts.
Microorganisms
Coliform spp
Pseudomonas spp
S. cereviceae
Z. baillii
L. plantarum
Oil
concentration
10 ppm
1.5
1
2
3
1.25
50 ppm
NI
1
2.5
1.5
1.5
100 ppm
2.5
1.5
3
2
2
NI: no inhibition
Plants are important source of potentially useful structures for new chemotherapeutic agents. The first
step towards this goal is the in vitro antibacterial activity assay (Abdel-Shafi, 2013). The E.
angustifolia EOs showed a weak activity against all the tested microorganisms, especially for
Pseudomonas spp, whose zones of inhibition ranged from 0 mm to 3 mm. The largest diameter (3
mm) was observed with 100 ppm of oils on S. cereviceae and 10 ppm on Z. baillii while the smallest
(no inhibition) was recorded with to 50 ppm of this extract on Coliform spp. Generally, yeasts are
more susceptible to the presence of EOs than the bacterial species confirming previous works
(Hammerschmidt et al., 1993; Charai et al., 1995; Hili et al., 1997; Nzeako et al., 2006; Mahboubi
and Kazempour, 2011). The reason why yeast is more susceptible to the extracts than bacteria is
unclear but it may be that at any given time, these oils may break up the structural integrity
of yeast faster than they dissociate bacteria (Nzeako et al., 2006). Among the test microorganisms, the
most resistant was Pseudomonas spp, which correlated with previous data (Hili et al., 1997;
Bergkvist, 2007). Pseudomonas is example of multiresistant bacteria that are becoming an alarming
problem within the healthcare system (Bergkvist, 2007). Nazzaro et al. (2013) indicated that this
resistance is due to the formation of exopolysaccharides that increase resistance to EOs. As shown in
Table, it was found that a Gram positive bacterium (L. plantarum) was slightly more susceptible than
Inhibitory Effects of Echinacea Angustifolia Essential Oils on the Growth of Five Pathogenic
Organisms: Coliform Spp, Pseudomonas Spp, Saccharomyces Cerevisiae, Zygosaccharomyces Bailii and
Lactobacillus Plantarum
International Journal of Advanced Research in Botany (IJARB) Page | 12
Gram negative bacteria (Pseudomonas spp). The weak antibacterial activity against the gram negative
bacteria was ascribed to the presence of an outer membrane which possessed hydrophilic
polysaccharides chains as a barrier for hydrophobic essential oils (Inouye et al., 2001). With increase
in concentration of essential oil, increase in zone of inhibition was observed thus dose-dependent
response was clear for essential oil, except Z. baillii.
As per the available literature, there is not much experimental evidence with regard to antimicrobial
activities on E. angustifolia extract. In contrast to above obtain results, Wendakoon et al. (2012)
reported that E. angustifolia extract did not show any antibacterial activity against S. aureus, S.
epidermidis, E. coli, P. aeruginosa and S. enteritidis. Also, Izzo et al. (1995) in his study to the
antibacterial activity of 68 plant extracts against eight bacteria (Bacillus subtilis, Staphylococcus
aureus, Streptococcus haemolyticus, Escherichia coli 7075, Klebsiella
Binns
pneumoniae, Proteus
mirabilis, Pseudomonas aeruginosa, Salmonella typhi Z-Z) reported that Extracts of E. angustifolia
D.C showed activity only against Bacillus subtilis.
Mir-Rashed et al. (2010) found that all Echinacea extracts tested had antifungal activity against the
wild type S. cerevisiae S288C. Whereas, Binns et al. (2000) reported that hexane extracts of
Echinacea variably inhibit growth of yeast strains of Saccharomyces cerevisiae, Candida shehata, C.
kefyr, C. albicans, C. steatulytica and C. tropicalis under near UV irradiation (phototoxicity) and to a
lower extent without irradiation (conventional antifungal activity). Sharma et al. (2008) had screened
and tested six different commercial Echinacea extracts for their antibacterial activity against 15
different human pathogenic bacteria and two pathogenic fungi. They observed that E. angustifolia
extracts exhibit strong growth inhibition against Haemophilus infuenzae, moderate activity against
Clostridium difficile and Legionella pneumophila but inactive on other microorganisms
(Propionibacterium acne, Acinetobacter baumanii, Bacillus cereus, Bacillus subtilis, Enterococcus
faecalis, Escherichia coli, Klebsiella pneumoniae, Mycobacterium smegmatis, Pseudomonas
aeruginosa, Candida albicans and Trichoderma viride).
Similarly, Bírošová et al. (2010) studied antimicrobial activity of extracts of underground and above-
ground parts of E. angustifolia against Mycobacterium smegmatis, Staphylococcus aureus,
Staphylococcus epidermidis, Escherichia coli and Salmonella Typhimurium, Alternaria alternata,
Aspergillus fumigatus, Microsporum gypseum and Trichophyton terrestre. Their results showed that
Radix extract had the highest antimicrobial activity both against bacteria and filamentous fungi and no
growth inhibition of all tested bacteria was observed at the extract from herba of E. angustifolia.
It was observed that the antimicrobial activity of plant extract varies from one plant to another in
different studies carried out in different parts of the world. The variation in results of different
researches may be due to many factors such as, the effect of climate, soil composition, age and
vegetation cycle stage, on the quality, quantity and composition of extracted product, different
bacterial strains and type of solvent used for extraction (Ababutain, 2011). The antimicrobial activity
has been attributed to the presence of some active constituents in the extracts (Joshi et al., 2011). The
chemical composition of various plant parts from the three Echinacea commonly used as medicines,
E. pallida var pallida, E. pallida var angustifolia and E. purpurea, is well established (Binns et al.,
2002). Three groups of compounds in these Echinacea species have pharmacological activity: the
caffeic acid derivatives (CADs), the lipophilic alkamides, and the highly polar polysaccharides
(Bauer, 2000; Clifford et al., 2002). In early research, Echinacoside - a caffeic acid derivative-
demonstrated weak antimicrobial activity against Staphylococcus aureus in vitro (Stoll et al., 1950).
The alkamides have shown strong inhibitory activity in vitro against Saccharomyces cerevisiae by
disruption to the fungal cell wall/membrane complex (Cruz et al., 2014). In immunodeficient mice,
treatment with E. purpurea polysaccharide led to enhanced production of TNF-a and enhanced
cytotoxicity against Leishmania enrietti, and protected the mice against lethal infections with Listeria
monocytogenes and Candida albicans (Goldhaber-Fiebert and Kemper, 1999). Some experts believe
that the polysaccharides are primary active ingredients for immune modulating effects (Tubaro et al.,
1987; Wagner et al., 1988). It appears that the immune-stimulating effects of Echinacea result from
polysaccharides surrounding tissue cells and thereby providing protection from bacterial and
pathogenic invasion (Newall et al., 1996).
Bachir Raho G et al.
International Journal of Advanced Research in Botany (IJARB) Page | 13
The observed low antimicrobial activity of E. angustifolia essential oil founded in our study could be
associated with the low amount of those active components.
4. CONCLUSION
This study has shown that essential oils from E. angustifolia, displayed inhibitory activity against the
tested microorganisms to varying degree, higher against L. plantarum than Pseudomonas spp. The
bioassay confirms that yeasts are more susceptible than the bacteria and Gram positive bacteria are
more sensitive compared to Gram negative ones, Pseudomonas spp being in general the most resistant
strain. Essential oils are potential agent against both bacteria and yeast. Similar experimentations can
help to explore the potential role of essential oils as antimicrobial agents but requires further study.
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