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ANTIMICROBIAL ACTIVITY OF FRESH PLANT JUICE ON THE GROWTH OF

BACTERIA AND YEASTS


Merih KIVAN
Dept. of Biology, Faculty of Science, Univ. of Anadolu, Eskiehir, TURKEY
Buket KUNDUHOLU
Dept. of Biology, Faculty of Science and Art, Univ. of Osmangazi, Eskiehir,
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.
ZET
Bu almada 17 deiik bitkiden elde edilen extrenin kltr ortamnda bakteri ve mayalar
zerindeki inhibitr etkisi test edilmitir. Bakteriler zerinde en yksek inhibitr aktivite
gsteren bitkinin sarmsak olduu belirlenmitir. Soan, prasa, krmz biber, bahe turpu ve
yaban turpu 1-3 bakteri tr zerinde inhibitr etki gstermitir. Dier bitki extreleri (lahana,
nane, dereotu ve maydanoz) test edilen bakterilerde inhibisyon oluturmamtr. Sarmsak,
soan (1 ve 2), dereotu ve lahana extreleri test edilen tm mayalarn geliimini inhibe
etmitir. Test edilen mayalar zerinde en dk inhibitr etkiye ise prasa, bahe turpu ve
nane extrelerinin sahip olduu belirlenmitir.
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). Kvan and Akgl (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 fr 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 Eskiehir, 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-peptoneglucose 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 horseradish (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
1. Zaika, L.L. Spices and herbs: their antimicrobial activity and its determination
J. Food Safety, 9:97-118 (1975)
2. Shelef, L.A., Naglik, O.A and Bogen, D.W. Sensitivity of some common
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)
4. Diaz, R., Quevedo-Sarmiento R. and.Ramos-Cormenzana, A. Phytochemical
and antibacterial screening of some spices of Spanish Lamiaceae Fitoterapia,
4,329-333, (1988)

5. zelik, S., Farkl


yrelerden alnan soan (Allium cepa L.) rneklerindeki
antimikrobiyal maddelerin (fitonsid) bakterisit ekilerinin aratrlmas , Gda,1
42-46 (1986)
6. Mantis, A.J., Karaioannoglou, P.G. and Spanos, G.P. et al. The effect of
garlic extract on food poisoning bacteria in culture media. I. Staphylococcus
aureus Lebensm. Wiss. Technol. 11:26-28 (1978)
7. Al-Delaimy, K. S.and Ali, S. H. Antibacterial action of vegetable extracts on
the growth of pathogenic bacteria J. Sci. Food Agric. 21:110-112 (1970)
8. Johnson, M. G. and Vaughn, R.H. Death of Salmonella typhimurium and
Escherichia coli in the presence of freshly reconstituted dehydrated garlic and
onion Appl. Microbiol., 17, 903-905 (1969)
9. Temmyo, R. Studies on the prevention of outbreaks of food poisoning caused
by Vibrio parahaemolyticus Bull. Tokyo Med. Dent. Univ. 13: 489 (1966)
10. Beuchat, L.R., Sensitivity of Vibrio parahaemolyticus to spice and organic
acids J. Food Sci., 41: 899-902 (1976)
11. Salzer, J.J., Broker U and H. F. Klie et al. Wirkung von pfeffer und
pfefferinhaltssteffen auf die microflora von wurtswaren Die Fleischwirtschaft
57:1975-1976 (1977)
12. Huhtanen, C.N., Inhibition of Clostridium botulinum by spice extracts and
aliphatic alcohols J. Food Protec. 43(3):195-196 (1980)
13. Moore, G.S. and Atkins, A.D. The fungicidal and fungistatic effects of an
aqueous extract on medically important yeast-like fungi Mycologia, 69:341348 (1977)
14. Tynecka, Z. and Gos, Z. The inhibitory action of garlic (Allium sativum L.)
on growth and respiration of microorganisms Acta Microbiologica Polanica.,
5B: 51-62 (1973)
15. Kvan , M. and A. Akgl, Mayalarn gelimesi zerine baharatlarn etkisi,
G da, 2, 145-152 (1988).
16. Kvan , M. and Akgl, A., Antibacterial activities of essential oils from
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)
18. Amonkar, S.V. and Banerji, A. Isolation and characterization of larvicidal
principle of garlic Science, 174:1343-1344 (1971)
19. Pruthi, J.S. Properties and uses In Spices and Condiments. Chemistry,
Microbiology and Technology, Academic Press, Inc. New York, pp. 16-68
(1980)
20. Altman, L.V., Fedotova, N.L and Babichenko,L.V. et al. Bactericidal
properties
of
freeze
and
dried
horse
radish
Konservnaya;
Ovoshchesushilnaya, Promyshlenost, 9, 40 (1980)
21. Conner, D.E. and Beuchat, L.R. Effects of essential oils from plants on
growth of food spoilage yeasts J. Food Sci., 49(2):429-434 (1984)

Table 1: The commercial and scientific names of plant tested.


Plan
t name

Botanical Source

Family

Plant Part
Garl

ic Allium sativum L.
Onion (3 cultivars)
Leek
Horse-radish (black)
Garden-radish
Horse-radish (white)
Cabbage
Brussels sprout
Cauliflower
Rocket plant
Peppermint
Celery
Red pepper
Dill
Parsley

Liliaceae
Allium cepa L.
Allium porrum Don.
Raphanus raphanistrum L.
R. sativus L. var. radicula
R. sativus L.
Brassica oleracea L. var. oleracea
B.oleracea L. var. gemmifera
B.oleracea L. var. botrytis
Eruca sativa Miller
Mentha spicata L.
Apium graveolens L.
Capsicum annum L.
Anethum graveolens L.
Petroselinum sativum

bulb
Liliaceae
Liliaceae
Cruciferae
Cruciferae
Cruciferae
Cruciferae
Cruciferae
Cruciferae
Cruciferae
Labiatae
Umbelliferae
Solanaceae
Umbelliferae
Umbelliferae

bulb
bulb
root
root
root
leaf
leaf
flower
leaf
leaf
bulb
fruit
leaf
fruit

Table 2: Tested bacteria and yeast


B
acteria

Strain no

Yeasts

Strain no
Ba

cillus cereus
Bacillus subtilis
Enterobacter aerogenes
Escherichia coli
Klebsiella pneumoni
Proteus vulgaris
Pseudomonas aeruginosa
Salmonella typhimurium
Serratia marcescens
Staphylococcus aureus
Vibrio parahaemolyticus

B 83
B 74
En 65
E9
Kl 31
Pr 14
Ps 1
S 498
Sm 4
St 14
V. 98

Candida crucei
Candida utilis
Candida tropicalis
Pichia membranafaciens
Rhodotorula rubra
Saccharomyces cerevisiae
Saccharomyces rouxii
Saccharomyces pombe
Saccharomyces octoporus
Shizosaccharomyces bailii

FRR 1302
FRR 1650
FRR 1282
FRR 1235
FRR 2406
FRR 1519
FRR 1911
FRR 2208
FRR 2171
FRR 2227

Table 3: Inhibitory properties (inhibition zone diameter in mm) of plant juice towards
bacteria.
Plan
t

Garlic

Onion
1

Onion
2

Onion
3

Leek

Red
pepper

Garden Horse
radish radish
white
B.ce

reus
B. subtilis
E. aerogenes
E. coli
K. pneumoni
P. vulgaris
P. aeruginosa
S. aureus
S. typhimurium
S. marcescens
V. parahaemolyticus

20
26
26
22
28
14
24
28
18

14
28
12
-

12
20
-

18
13
-

28
-

28
30
16
-

13
24
-

12
-

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

Garlic

Oni.

Oni.

Oni.

Leek Horse

Horse

rad.

rad.

black

white

10
28
11
16
-

10
12
13
12
28
-

Garden Cabbage
rad.

Pepper Red Dill


mint

pep.

14
17
10

15
24
11
21
10
-

Parsley

a
b
c
d
e
f
g
h
i
j

17
38
22
23
17
23
26
15
26
32

18
20
28
19
24
23
15
26
28
28

20
38
28
16
25
10
20
10
10
28

20
13
16
28
18
10
38
26
26

10
-

ND
ND
ND
ND
ND
ND
ND
10
12
28

28
13
18
15
13
19
14
-

17
32
28
19
13
18
17
28
26
32

14
10
13
10
18
-

(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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