UJMR, Volume 2 Number 1 June, 2017

Received: 9th Dec, 2017 Accepted: 23rd Feb, 2017

Antibacterial activity of Phoenix dactylifera L. (Date palm) Seeds Extract

against Escherichia coli
* Ado, A., Chonoko, U. G., and Abdul- Azeez, U. M.
Department of Applied Science, College of Science and Technology, Kaduna Polytechnic,
Kaduna,Nigeria
* Correspondence author: [email protected]

Abstract
This research determines antibacterial activity of Phoenix dactylifera against clinical isolate of
Escherichia coli. Extraction, phytochemical screening and agar diffusion methods were
employed to evaluate phytochemical profile, and antibacterial activity of Phoenix dactylifera.
Alkaloids, cardiac glycosides and saponnins are present in Phoenix dactylifera seeds. Phoenix
dactylifera seeds showed activity against E. coli with zone of inhibition of 20.4mm. The
Minimum inhibitory concentration (MIC) and Minimum bactericidal concentration (MBC) of
Phoenix dactylifera seed extract are 1000µg/ml and 100µg/ml against E. coli respectively.
Phoenix dactylifera seed could be very useful in the management of gastrointestinal infection
due to E. coli.
Key words: Antibacterial, Phoenix dactylifera, phytochemical, Extraction, gastrointestinal,
Eschrichia coli

INTRODUCTION and Marshall, 2003). People with weakened

Phoenix dactylifera (Date palm) is a flowering immune systems, pregnant women, young
plant species belonging to the family Arcaeae. children, and old adults are at increased risk
It is cultivated for edible sweet fruits and is for developing these complications. Most
integrated as part of Arabian diet as a good intestinal infections are caused by
source of low cost food (Abdelhak et al., 2005). contaminated food or water (Al-Shahib and
It is a medium sized plant, 15-25m tall, growing Marshall, 2003).
with a single root system; the leaves are 4-6cm Dates provide essential nutrients and are
long.Date palm fruit is found useful in the claimed to have benefits in human health.
treatment of inflammation, fever, paralysis, Manickarasagan et al., (2012) reported that
nervous disorders and memory disturbances (Al- dates by-products can be considered a good
Qarawi et al.,2004). It is used as astringent in source of dietary fibre, total phenolics and an
intestinal troubles, treatment of sorethroats, inexpensive source of natural antioxidants and
colds, bronchial catarrh, fever, gonorrhea, could be used as a functional food ingredient.
edema liver and abdominal troubles, and to This study aimed to evaluate the antibacterial
counteract alcohol intoxication (Al-dalhan and activity of Phoenix dactylifera ethanol seed
Bhat,2012). extract on Escherichia coli.
Date palm seed powder is used in traditional
medicines to treat toothache (Morton, MATERIALS AND METHODS
1987).Dates seed extract has ability to restore Sample collection and handling
the normal function status of the poisoned Dried Phoenix dactylifera (Date) fruits were
liver, and was found protective against carbon purchased in March, 2016 from Central Market,
tetrachloride hyper toxicity on the liver in rats Kaduna. The sample of the Phoenix dactylifera
(Al-Qarawi et al., 2004). Date seeds have been fruits were identified and authenticated using
studied as potential sources of edible oils and Voucher specimens with reference number
pharmaceuticals (Al-Shahib and Marshall, 2003). 040616 deposited in the herbarium of Applied
E. coli is a gram-negative, facultative Science Department, Kaduna Polytechnic,
anaerobic, rod-shaped bacterium, commonly Kaduna. The seed of the dried Phoenix
found in the warm blooded organisms. E. coli is dactylifera (Date) fruits were removed and
a common cause of intestinal infections. ground using mechanical grinder until a
Symptoms of intestinal infection include powdery texture was achieved. The powdered
diarrhea, abdominal pain, and fever. More sample was transferred into clean plastic
severe cases can lead to bloody diarrhea, container and kept at room temperature (28 ±
dehydration, or even kidney failure (Al-Shahib 2oC) prior to extraction.

6
UMYU Journal of Microbiology Research
 UJMR, Volume 2 Number 1 June, 2017

Ethanol solvent extraction of Phoenix Preparation of varied concentration of

dactylifera seed was carryout using soxhlet Phoenix dactylifera extracts
extractor. The extract was separated from the Two grams (2g) of the ethanolic extract of
solvent using solvent recovery method. The Phoenix dactylifera was weighed using the
recovered extract was stored at room analytical weighing balance and was dissolved
temperature. in 20ml of sterile distilled water. It was mixed
thoroughly until dissolved solution was
Phytochemical screenng obtained. This formed the stock solution. The
The ethanolic extract of Phoenix dactylifera stock solution was used to prepare 10, 100 and
seed was phytochemically screened for the 1000µg/ml by diluting with sterile distilled
presence of alkaloids, tannins, steroids, water, using 1 milliliter sterile syringe, after
saponins, and cardiac glycosides using the calculating the required diluents volume using
method reported by (Sofowora, 1993). dilution formula (Ademuyiwa et al., 1990).

Clinical bacterial isolates Antibacterial activity of Phoenix dactylifera

Clinical isolates of Escherichia coli cultures seed extract
were obtained from Shehu Kangiwa Medical The agar well diffusion method was used to
Centre (SKMC), Kaduna Polytechnic. These determine the antibacterial activity of P.
isolates were confirmed using Grams staining dactylifera seed extract. The sterile nutrient
and biochemical tests (Cheesbrough, 2004). agar was poured in sterile Petri plates and
allowed to solidify. Sterile wire loop was used
Preparation of culture media to inoculate standardized bacterial inocula
Nutrient agar and nutrient broth were prepared radially on the surface of the prepared nutrient
according to manufacturer’s instructions and agar. A sterile standard cork-borer (6mm) was
autoclaved at 1210C for 15minutes. used to bore wells on the surface of the agar.
Using a sterile syringe, 0.1ml of the different
Preparation of over night broth culture extract were saperately dispensed in to the
Two to three well grown colonies from each of wells and allowed to stand for 20minutes at
the stored slant cultures were separately and room temperature for proper diffusion of
aseptically introduced into the sterile nutrient extract. The preparations were incubated at
broth in test tubes. These were incubated at 370C for 24 hours. Zones of inhibitions were
370C for 24 hours. measured in millimeters using meter rule.

Preparation of 0.5 McFarland turbidity Determination of Minimum Inhibitory

standards Concentration (MIC)
A 1%v/v solution of sulphuric acid (H2SO4) was The tube dilution method was used as
prepared by adding 1ml of concentrated H2SO4 described by (Pelzer et al.,1999). Standardized
to 99ml of water in a volumetric flask. The suspension of the test organisms were
preparation was properly mixed. A1% inoculated into series of test tubes containing
solution of barium chloride was prepared by sterile nutrient broth.
dissolving 0.5g of dehydrated barium chloride Varied concentrations of the extract were
(BaCl2.2H2O) in 50ml of distilled water in a sequentially introduced into the inoculated test
volumetric flask. A 0.6ml of the barium tubes.
chloride solution was added to 99.4ml of the The preparations were incubated at 370C for 24
H2SO4 solution and mixed properly. A small hours. Tubes without turbidity were recorded
volume of the turbid standard solution was put as MIC.
into a screw cap bottle. The standard was kept
at room temperature (28±20C) prior to inocula Determination of Minimum Bactericidal
standardization (Cheesbrough, 2000). Concentration (MBC)
The minimum bactericidal concentration was
Standardization of inocula determined by sub-culturing the minimum
The previously prepared overnight broth inhibitory concentration tubes that showed no
cultures of each bacterial isolate was adjusted growth on nutrient agar and incubating for
to 0.5 McFarland standard. This was achieved 24hours at 370C. The minimum bactericidal
by adding sterile saline solution to each broth concentration was represented by the plate
culture till the turbidity matched standard with the lowest concentration without growth
(Hussein et al., 1998). (Roberts et al., 2000).

7
UMYU Journal of Microbiology Research
 UJMR, Volume 2 Number 1 June, 2017

RESULTS phytochemical screening of Phoenix dactylifera

Extraction of phoenix dactylifera seeds yields seeds extract is presented in Table 3. Alkaloid,
32.8% of extract (Table 1). The result of Gram’s Cardiac glycosides and Saponins are present in
reaction presented in Table 2 confirms E. coli ethanol seed extract of Phoenix dactylifera
as Gram negative and is positive when tested while Steroids and Tannins are absent.
for indole, motility and citrate utilization. The

Table 1: Percentage yield of Phoenix dacylifera seed extract

Extract Initial weight of Weight of Yield (%)
sample (g) extract (g)
Ethanolic seed Extract 25 8.2 32.8

Table 2: Gram reaction and biochemical characteristics of clinical bacterial isolates

Isolates Gram reaction Indole Motility Citrate
E. coli - + + +

Table 3: Phytochemical profiles of Phoenix dactylifera ethanol seed extract

Phytochemical profiles
Extract Alkaloids Cardiac glycosides Saponnins Steroids Tannins
P. dactylifera + + + - -
seed extract
KEY: + Means presence of phytochemical, - Means absent of phytochemical

Antibacterial activity of ethanolic extract of 4. The seed extract was found effective at
date seeds 1000µg/ml as against Escherichia coli as it
The antibacterial activity of Phoenix demonstrates higher zone of inhibition of
dactylifera seed extract is presented in Table 20.4mm.

Table 4: Zones of inhibition (mm) of ethanolic extract of date seeds against E. coli
Zones of inhibition (mm)
Extract 10µg/ml 100µg/ml 1000µg/ml Ciprofloxacin (30μg/ml)
Escherichia 13mm 15mm 20.4mm 25mm
coli
Table 4: MIC and MBC of Phoenix dactylifera seed extract against Escherichia coli
Test Organism MIC (µg/ml) MBC (µg/ml)
Escherichia coli 1000 1000

DISCUSSION AND CONCLUSION Furthermore, it has been proposed that its

This study investigates antibacterial effect of antioxidant constituents account for its
Phoenix dactylifera ethanolic seeds extract beneficial therapeutic effects (Ljubuncic et al.,
against Escherichia coli. Many plants in 2005). The antioxidant activity of phenolic
different location have been recognized as a compounds is a result of their redox properties,
source of cure for ailments in their region of which can play an important role in absorbing
existence. The plant parts mostly used include and neutralizing free radicals, quenching
seed, back, leaves (Muktar and Tukur, 1999). singlet and triplet oxygen, or decomposing
Dates have medicinal uses including anticancer, peroxides (Farag, 2011).
antihyperlipidemic, hepatoprotective activities The presence of phytochemicals such as
and thereby serving as an essential healthy Alkaloids, Cardiac glycoside and saponins in
food in the human diet (Biglari et al., 2009). Phoenix dactylifera reveals the efficacy of seed
The date fruit is used in folk medicine to treat extract against the bacterial isolate. (Habib
the different infectious diseases probably and Ibrahim, 2009). Saponins are harmless in
because of their antibacterial ability, the body when taken orally (Fatope, 1994). The
immuneomodulatory activity and antifungal absence of tannins among the pthytochemical
property (Baliga et al., 2011). components detected in this study is probably
The medicinal use of extracts prepared from as a result of drying process of the Phoenix
plant parts of the dates back to ancient times. dactylifera.

UMYU Journal of Microbiology Research 8

 UJMR, Volume 2 Number 1 June, 2017
Maillard and Berset, (1995) reported that in the Escherichia coli. The Minimum inhibitory
process of drying, tannins are degraded by heat Concentration (MIC) and Minimum bactericidal
and maturation enzymes which result in release Concentration (MBC) will go a long way in
of phenolic compounds. Escherichia coli can be providing therapeutic basis of Phoenix
transmitted from person to product through dactylifera seeds in treating bacterial
unhygienic practice such as handling products Infections due to Escherichia coli (Al-Farsi and
with infected hands (Sawaya et al., 2006). The Lee, 2008).
extract has varying degree of antibacterial
activities against Escherichia coli. The high CONCLUSION
concentration of the Phoenix dactylifera seed Phoenix dactylifera seeds extract has
extract was found effective and thus useful in antibacterial effect against Escherichia coli due
the control of the bacterial infection due to to abundance of phytochemical in the seeds.

REFERENCES Fatope, M. O. (1994). Phytocompounds: their

Abdelhak, M., Guendez, E., Eugene, K. and bioassay and diversity and innovation,
Kefalas, P. (2005). Phenolic profile and 7:229-236.
antioxidant activity of the Algerian ripe Habib, H. M. and Ibrahim, W. H. (2009).
date palm fruit (Pheonix dactylifera). Food Nutritional quality evaluation of eighteen
Chemistry, 89:411-420. date pit varieties. International Journal of
Ademuyiwa, O., Ngaha, E. O. and Ubah, F. O. Food Science and Nutrition, 60:99-111.
(1990). Vitamin E and Selenium in Hussein, S. M., Hossain, M. M, Faruque, A.,
Gentamicin mephrotoxicity. Human and Mazumder, M. E. H., Rana, M. S., Akter, R
Experimental Toxicology, 9:281-288. and Alam, M. A. (1998). Comparison of
Al-Dalhan, S. and Bhat, R. S. (2012). Antibacterial antioxidant potential of different fractions
activities of extracts of leaf, fruit, seed and of Commelina benghalensis. Linn Bang.
bark of Pheonix dactylifera. African Journal Journal of Life Science. 20(2):9-16.
of Biotechnology. 11(24):10021-10025. Ljubuncic, P., Portnaya, I., Cogan, U., Azaizeh, H.
Al-Farsi, M. A. and Lee, C. Y. (2008). Nutritional and Bomzon, A. (2005). Antioxidant
and functional properties of dates: a review activity of Crataegus aronia aqueous extract used
critical review in food science and in traditional Arab medicine in Israel.
nutrition, 48:877-887. Journal of Ethnopharmacol.,101: 153-61.
Al-Qarawi, A. A., Mousq, H. M., Beh, B., Abdel- Maillard, M. N., and Berset, C. (1995). Phenolic
Rahman, H. and El-Mougy, A. A. (2004). profile and antioxidant activity of the
Traditional medicine in Africa. Algerian ripe date palm fruit (Phoenix
International Journal of Research dactylifera). Journal of Food Chemistry;
Veterinary Medicine. 2:176, 242-251. 89: 411−420.
Al-Shahib, W. and Marshall, R. J. (2003). Fatty Manickavasagan, A., Essa, M. M. and Sukumar, E.
acid content of the seeds from 14 varieties (2012). Dates: production, processing, food
of dated palm Pheonix dactylifera L. and medicinal values CRC press pp415.
International Journal of Food Science Morton, F. (1987). In: fruits of warm climates.
Technology. 38:709-712. Julia F. Morton Miami, 5-11.
Baliga, S, Baliga, V. and Kandathil S. (2011). A Muktar, M. O. and Tukur, A. (1999). In-vitro
review of the chemistry and pharmacology screening for activity of Pistia stratiotres
of the date fruits (Phoenix dactylifera L.). extracts NISEB Journal 1(1): 51-60.
Journal of Food Research International, Pelzer, M. S., Chem, S. S. and Kriey N. R. (1999).
44: 1812-22. Microbiology 5th Edition Mc Graw – Hill.
Biglari, F., AlKarkhi, A. F. M. and Mat, E, A. Companies, New York, Delhi Pp. 261-
(2009). Cluster analysis of antioxidant 280,160-170.
compounds in dates (Phoenix dactylifera): Roberts, D., Veeramachaneni, D. N., Sehlaff, D.
Effect of longterm cold storage. Journal of and Awoniyi, C. A. (2000). Effects of
Food Chemistry, 998–1001. chronic dietary exposure to genistein, a
Cheesbrough, M. (2004). District Laboratory phytoestrogen during various stages of
Practices in Tropical Countries part 2 (2nd development on reproduction and
edition). Cambridge University Press, spermatogenesis in rate endocrine, 13,281-
United Kingdom 64-68, 70 – 103, 131-148; 286.
141-222. Sawaya, W. N., Miski, A. M., Khalechadourian, A.
Farag, A. A. (2011). Effect of Dates During the A. and Mashadi, A. S. (1983). Medicinal
Tapering Period on Some values of Date Palm. Journal of Food
Antioxidants,Glucose and Record Level for Science and Nutrition. 2:1-25.
50m Freestyle Young Swimmers; World Sofowora, A. (1993). Medicinal plants and
Journal of Sport Science, 3 (201): 1106- traditional medicine in Africa. Spectrum
1110. books Ltd, Ibadan, Nigeria, pp191-289.

UMYU Journal of Microbiology Research 9