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Antimicrobial activity of fresh plant juice on the growth of bacteria and

yeasts

Article · January 1997

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Merih Kivanc Buket Kunduhoğlu

Anadolu University Eskisehir Osmangazi University
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 ANTIMICROBIAL ACTIVITY OF FRESH PLANT JUICE ON THE GROWTH OF
BACTERIA AND YEASTS

Merih KIVANÇ
Dept. of Biology, Faculty of Science, Univ. of Anadolu, Eskişehir, TURKEY
Buket KUNDUHOĞLU
Dept. of Biology, Faculty of Science and Art, Univ. of Osmangazi, Eskişehir,
TURKEY.

SUMMARY
In this study fresh juice of 17 plants were prepared and tested for inhibition of bacteria and
yeasts in culture media. Garlic was the most inhibitory plant on the bacteria. Onion, leek, red
pepper, garden radish and horse radish inhibited the growth of one to three bacteria. Other
plant juice did not inhibit the growth of tested bacteria. Garlic, onion, dill and cabbage juice
inhibited the growth of all 10 yeasts. Leek and peppermint juice were the least inhibiting
plants on the tested yeasts.

ÖZET
Bu çalışmada 17 değişik bitkiden elde edilen extrenin kültür ortamında bakteri ve mayalar
üzerindeki inhibitör etkisi test edilmiştir. Bakteriler üzerinde en yüksek inhibitör aktivite
gösteren bitkinin sarmısak olduğu belirlenmiştir. Soğan, pırasa, kırmızı biber, bahçe turpu ve
yaban turpu 1-3 bakteri türü üzerinde inhibitör etki göstermiştir. Diğer bitki extreleri (lahana,
nane, dereotu ve maydanoz) test edilen bakterilerde inhibisyon oluşturmamıştır. Sarmısak,
soğan (1 ve 2), dereotu ve lahana extreleri test edilen tüm mayaların gelişimini inhibe
etmiştir. Test edilen mayalar üzerinde en düşük inhibitör etkiye ise pırasa, bahçe turpu ve
nane extrelerinin sahip olduğu belirlenmiştir.

INTRODUCTION
Many of the plants used today were known to the people of ancient cultures throughout the
world and they were valued their preservative and medicinal powers. Scientific experiments
on the antimicrobial properties of plants and their components have been documented in the
th
late 19 century (1).
Naturally occurring microbial inhibitors have been recovered from a wide variety of foods
including onions, garlic, fruits, vegetables, cereals and spices. many of these antimicrobials
contribute to the food stuffs natural resistance to deteriorations. The flavour components
consist of such compounds as alcohols, aldehydes, esters, terpens, phenols, organic acids and
others, some of which have not yet been identified (2).
The widespread use of onion and garlic as a flouvering agent is well known. Onion and
garlic also known to have medicinal properties. Onion bulbs contain tannin, pectin, quercetin
and glycosides. Alcohol extract show hypotensive analgesic and antileptazol properties.
Onion extracts shows antibacterial properties. Garlic bulbs contain pectin, garlicin, volatile
oils, allin and allistatins I and II. Alcoholic extract of garlic shows bactericidal, antibiotic,
high hypoglycaemic and fungusidal activities. The extract showed hypotensive, analgesic
sedative and antileptazol properties (3).
Lamiaceae (Labiatae) are reputed for their medicinal uses. Due to their content in essential
oils, several species of this family show antimicrobial activity. Investigations on plants were
utilized in folk medicine and demonstrate the antimicrobial activity. Several species of
Mentha (ethanol and ethyl ether extract) show antimicrobial activity on E. coli, M. luteus and
B. subtilis. It concerns volatile oils, catechic tannins, flauonoids and anthraquinones (4).
Celery bulbs contains etheric oils, and red pepper contains alcoholoids (capsaicin) (3).
Recent studies have shown antimicrobial activity of onion and garlic (5,6). Al Delaimy and
Ali (7) reported that filtered onion extracts were bactericidal against Shigella dysenteria and
Staphylococcus aureus. Onion and garlic powders inhibited growth of Salmonella
typhimurium and Escherichia coli (8). Vegetable extracts examined for their suitability to
support the growth of Vibrio parahaemolyticus, eggeble, tomato, carrot, parsley, celery and
onion were shown to be inhibitory (9). Beuchat (10) studied the effect of horse radish, onion
powder, garlic powder, paprika on the growth of the V. parahaemolyticus.Pepper was active
against E. coli in sausage according to Salzer et al (11). Huhtanen (12) showed paprika and
red pepper to be active against Clostridium botulinum.
Moore and Atkins (13) reported that 22 active pathogenic isolates of Candida albicans
were all inhibited by the garlic extract. Candida albicans was inhibited by plant extracts,
including those from garlic (14). Kıvanç and Akgül (15) studied the effect of spice on the
growth of yeasts.
The objective of this study were determine the effects of fresh vegetable juice on
inactivation of selected microorganisms in laboratory media.

MATERIALS AND METHODS

Microorganisms and inocula:
Eleven bacteria and ten yeast were tested (Table 1). Lyophylized bacteria cultures obtained
from Bundesanstalt für Fleischforschung (Kulmbach, FRG). Ten yeasts were supplied from
CSRIO, Food Research Division (North Ryde, Australia).
Stock cultures of Vibrio parahaemolyticus were maintained tryptic soy agar (which contain
3 % NaCl) and the other bacteria cultures were maintened on nutrient agar. And yeast stock
cultures were maintened on malt extract agar and kept at 4° C (2). Inocula for all experiments
were prepared by transferring a loop full of stock cultures to tubes and incubating at 35° C for
18-24 h (bacteria) and 30° C for 48 h (yeasts), respectively.
Fresh vegetable juice:
Seventeen vegetables were purchased from a local market in Eskişehir, TURKEY (Table
2). The whole samples purchased for each vegetable were washed with tap water and sterile
water respectively. Each vegetable was grated and pressed. Vegetable juice were sterilized by
filtration. These liquid was used for testing inhibitory activity.
Inhibitory activity was tested on nutrient agar and yeast extract-malt extract-peptone-
glucose agar (YMPG)(16) for bacteria and yeasts, respectively. For yeasts, the medium was
adjusted to pH 5.5 with 6N HCl (2,16)
For the growth experiments, 0.1 ml of the original cultures (about 106-107 cells) were
added into sterile duplicate sets of petri dishes and 20 portions of the molten (45° C) agar
were poured into petri dishes. The fresh vegetable juice (0.1ml) were placed in wells (8mm
diameter) cut in the agar media and plates were incubated at 37° C in the case of bacteria and
at 30° C in the case of yeasts (16). The resulting inhibition zones obtained with bacteria and
yeasts were measured after 48 h.
All experiments were done in two replicates and with control plates.
RESULTS AND DISCUSSION
The results obtained in the experiments are shown in Table 3. Fresh garlic juice showed the
highest antibacterial activity. This observation confirms an earlier report (5-7). Saleem and Al
Delaimy (7,17) reported that 3%, 5% and 10% fresh garlic extract inhibited the growth of B.
cereus on nutrient agar plates by 31.3%, 58.2% and 100%, respectively. The inhibition of B.
cereus increased with rising concentrations of garlic extract. Garlic juice was not inhibited B.
subtilis and P. aeruginosa. In garlic, the active inhibitory principal is allicin (diallyl disulfide
and diallyl trisulfide) (18,19). Allicin is enzymatically released from precursor after the garlic
bulbs are crushed (17).
The lowest effect was obtained with horse-radish and leek juice which inhibited the growth
of B. cereus and B. subtilis, respectively. Garden-radish, red pepper and three varieties of
onions inhibited the growth of two or three bacteria. The low activity of onion juice was
somewhat surprising. Beuchat (10) reported that V. parahaemolyticus was grown in onion
powder. Al-Delaimy and Ali (7) reported that 4% of fresh onion extract were bacteriostatic
against E. coli and S typhosa.
Horse-radish, Brussels sprout, cabbage, cauliflower, peppermint, rocket plant, celery, dill
and parsley did not inhibit the growth of tested bacteria. Altman et al. (20) found that horse
radish were bactericidal against S. aureus.
B. cereus was the most sensitive of the tested bacteria, followed by E. aerogenes and S.
marcescens.
Inhibitory activities of fresh vegetable juices against yeasts are shown in Table 4. The fresh
vegetable juices caused different inhibition zones on the tested yeasts. The most active juice
were onion, garlic and dill. These inhibited growth of all yeasts.
Moore and Atkins (13) reported that 22 active pathogenic isolates of C. albicans were all
inhibited by garlic extract. C. albicans were all inhibited by plant extracts (18). The horse-
radish (black and white), cabbage, red pepper and parsley were fairly active. The lowest effect
was obtained with brussels sprout, cauliflower, rocket plant and celery which did not inhibit
the growth of tested yeasts. Conner and Beuchat (21) showed that of 32 different plant
essential oils, garlic, onion were particularly inhibitory to selected food spoilage and
industrial yeasts.
Comparison of the sensitivity of the tested yeasts towards plant juice showed that S.
cerevisiae and R. rubra were the most sensitive, while Candida was the most resistant. On the
other hand, Saccharomyces were sensitive to most juices but to varying degrees.
In general, plant juices were more active against yeasts then bacteria.

REFERENCES

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J. Food Safety, 9:97-118 (1975)
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food-borne bacteria to the spices sage, rosemary and allspice” J. Food Sci. 45:
1042-1044 (1980)
3. Adesina, S.K. “Studies on some plants used as anticonvulsants in Amerindian
and African traditional medicine” Fitoterapia, 5-6:147-162 (1982)
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42-46 (1986)
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acids” J. Food Sci., 41: 899-902 (1976)
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pfefferinhaltssteffen auf die microflora von wurtswaren” Die Fleischwirtschaft
57:1975-1976 (1977)
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13. Moore, G.S. and Atkins, A.D. “The fungicidal and fungistatic effects of an
aqueous extract on medically important yeast-like fungi” Mycologia, 69:341-
348 (1977)
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on growth and respiration of microorganisms” Acta Microbiologica Polanica.,
5B: 51-62 (1973)
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Gı da, 2, 145-152 (1988).
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Turkish spices and citrus” Flavour and Fragrance Journal, 1, 175-179 (1986)
17. Saleem, Z.M. and Al Delaimy, K.S. “Inhibition of Bacillus cereus by garlic
extracts” J. Food Protec., 45(11): 1007-1009 (1982)
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(1980)
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Ovoshchesushilnaya, Promyshlenost, 9, 40 (1980)
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Table 1: The commercial and scientific names of plant tested.

Plan
t name Botanical Source Family Plant Part

Garl
ic Allium sativum L. Liliaceae bulb
Onion (3 cultivars) Allium cepa L. Liliaceae bulb
Leek Allium porrum Don. Liliaceae bulb
Horse-radish (black) Raphanus raphanistrum L. Cruciferae root
Garden-radish R. sativus L. var. radicula Cruciferae root
Horse-radish (white) R. sativus L. Cruciferae root
Cabbage Brassica oleracea L. var. oleracea Cruciferae leaf
Brussels sprout B.oleracea L. var. gemmifera Cruciferae leaf
Cauliflower B.oleracea L. var. botrytis Cruciferae flower
Rocket plant Eruca sativa Miller Cruciferae leaf
Peppermint Mentha spicata L. Labiatae leaf
Celery Apium graveolens L. Umbelliferae bulb
Red pepper Capsicum annum L. Solanaceae fruit
Dill Anethum graveolens L. Umbelliferae leaf
Parsley Petroselinum sativum Umbelliferae fruit

Table 2: Tested bacteria and yeast

B
acteria Strain no Yeasts Strain no
Ba
cillus cereus B 83 Candida crucei FRR 1302
Bacillus subtilis B 74 Candida utilis FRR 1650
Enterobacter aerogenes En 65 Candida tropicalis FRR 1282
Escherichia coli E9 Pichia membranafaciens FRR 1235
Klebsiella pneumoni Kl 31 Rhodotorula rubra FRR 2406
Proteus vulgaris Pr 14 Saccharomyces cerevisiae FRR 1519
Pseudomonas aeruginosa Ps 1 Saccharomyces rouxii FRR 1911
Salmonella typhimurium S 498 Saccharomyces pombe FRR 2208
Serratia marcescens Sm 4 Saccharomyces octoporus FRR 2171
Staphylococcus aureus St 14 Shizosaccharomyces bailii FRR 2227
Vibrio parahaemolyticus V. 98
 Table 3: Inhibitory properties (inhibition zone diameter in mm) of plant juice towards
bacteria.

Plan
t Garlic Onion Onion Onion Leek Red Garden Horse
1 2 3 pepper radish radish
white

B.ce
reus 20 14 - 18 - - 13 12
B. subtilis - 28 - - 28 - - -
E. aerogenes 26 12 12 13 - - - -
E. coli 26 - - - - - - -
K. pneumoni 22 - - - - - - -
P. vulgaris 28 - - - - 28 - -
P. aeruginosa - - - - - 30 - -
S. aureus 14 - - - - - - -
S. typhimurium 24 - - - - - - -
S. marcescens 28 - 20 - - 16 24 -
V. parahaemolyticus 18 - - - - - - -

Table 4: Inhibitory properties ( inhibition zone diameter in mm) of plant juice towards yeasts

Garlic Oni. Oni. Oni. Leek Horse Horse Garden Cabbage Pepper Red Dill Parsley
1 2 3 rad. rad. rad. mint pep.
black white
a
17 18 20 20 - 10 - ND - - 15 17 -
b 38 20 38 13 10 - - ND 28 - 24 32 -
c 22 28 28 16 - - 10 ND 13 - - 28 -
d 23 19 16 28 - 28 - ND 18 - 11 19 14
e 17 24 25 18 - 11 12 ND 15 - 21 13 10
f 23 23 10 - - - 13 ND 13 - 10 18 13
g 26 15 20 10 - 16 12 ND 19 14 - 17 10
h 15 26 10 38 - - 28 10 14 - - 28 18
i 26 28 10 26 - - - 12 - 17 - 26 -
j 32 28 28 26 - - - 28 - 10 - 32 -

(a: C. crucei, b: C. utilis, c: C. tropicalis, d: P. membrafaciens, e: R. rubra, f: S. bailii, g: S. cerevisiae, h: S.

octoporus, i: S. pombe, j: S. rouxii).
ND: not determined
-: no inhibition zone

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