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Letters in Applied Microbiology 1989,8, 123-125 CSG

The bactericidal activity of tea and coffee

SHIMAMURA Department
* of Bacteriology and Immunology, Showa University School of
Medicine, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142, Japan

Received 22 August 1988 and accepted 21 October 1988

TODA,M., O K U B OS,. , HIYOSHI,R . & S H I M A M U RT. A ,1989. The bactericidal

activity of tea and coffee. Letters in Applied Microbiology 8, 123-125.
Extracts of black tea, green tea, pu-erh tea or coffee inhibited the growth of various
bacteria known to cause diarrhoeal diseases. Tea or coffee also showed bactericidal
activity against Staphylococcus aureus and Vibrio parahaemolyticus.

During the preliminary phase of a programme Miiller-Hinton agar (BBL) in petri dishes.
for the formulation of foods to aid the preven- Wells (8 mm diameter) were punched in- the
tion of cholera and other diarrhoeal diseases it agar using a stainless steel borer and were filled
was found, unexpectedly, that green tea agglu- with 0.1 ml of the tea or coffee extracts. The
tinated cholera vibrios and rendered them non- plates were incubated overnight at 37C and the
mobile. The potential bactericidal activity of diameter of any resulting zones of inhibition
three tea extracts and a single coffee extract was measured. Each combination of bacterial strain
investigated using a wider range of bacteria. and extract was repeated three times.

Materials and Methods VIABLE CELL C O U N T METHOD

BACTERIA Bacterial suspensions were diluted 1 : lo5 with

nutrient broth (2 times conc.). Equal volumes
The bacterial strains are listed in Table 1.
(1 ml) of the dilution and extracts of tea or
coffee were mixed and incubated at 37C and, at
P R E P A R A T I O N OF E X T R A C T S O F intervals, 0.1 ml of the mixture was spread on to
TEA OR COFFEE two separate nutrient agar plates which were
Tea leaves and coffee beans were obtained com- incubated overnight at 37C. The mean number
mercially. Tea leaves or powdered coffee were of colonies was enumerated and compared with
suspended in phosphate-buffered saline (PBS) at that of a control (PBS). This experiment was
20% or 25% (w/v) concentration, respectively. repeated four times for each combination of
The suspensions were held at room temperature extract and bacterial strain.
for 3 h and then centrifuged at 15000 rev/min
for 10min. The pH of the supernatant fluids was Results and Discussion
adjusted to 7.0 with 2.5 mol/l NaOH or
2.7 mol/l HCI. Extracts of tea and coffee were first examined
for the inhibition of bacterial growth of 24
strains of pathogens causing diarrhoeal diseases
by an agar diffusion method (Mitsuhashi et nl.
Bacterial suspensions were cultured in peptone 1981). The data are shown in Table 1. Black tea,
water for 18 h and 0.2 ml mixed with 10 ml of green tea, pu-erh tea (Chinese tea) and coffee
inhibited the growth of both Gram-positive and
* Corresponding author. negative bacteria. Gram-positive bacteria were
124 Masako Toda et al.
Table 1. Inhibition of bacterial growth by tea and coffee
Black Green Pu-erh
Bacteria tea tea tea Coffee
Staphylococcus aureus 209 P +++ +++
aureus ATCC 25923 +++ ++
epidermidis ATCC 1228 +++ +++
Salmonella typhi ++ +
typhimurium TSA-2121 ++ +
enteritidis 87-350 ++ -
Shigella Jexneri 2a + -
dysenteriae - ++
Enteroinvasive Escherichia coli
(EIEC) EI-4 0124:H- - -
Enterohaemorrhagic Escherichia coli
(EHEC) TC-91 0157:H7 - -
Enteropathogenic Escherichia coli
(EPEC) EP-01 01 11 - -
Enterotoxic Escherichia coli
(ETEC) F 86352-1 - -
Vibrio cholerae 0-1, V86 El Tor + ++
cholerae 0-1, 569B classical + +
cholerae non 0-1 ++ ++
parahaemolyticus +- +-
Juoialis CDC 9554-78
mimicus ATCC 33653 ++ ++
Plesiomonas shigelloides 83-640 ++ +-+
Aeromonas sobria TAE-217 ++
- -
hydrophila TAE-219
Yersinia enterocolitica TE-14 -
Pseudomonas aeruginosa
Enterobacter cloacae ATCC 23355 - -

+ + +, D > 1 6 mm; + +, 16 mm>D>12mm; +, 12 m m > D > 8 mm; -, no


more sensitive than Gram-negative bacteria and The components of tea and coffee responsible
tea was a more effective inhibitor than coffee. for the observed bactericidal activity have yet to
Staphylococcus aureus, Staph. epidermidis, Vibrio be identified. The tea and coffee samples tested
cholerae non 0-1 and Plesiomonas shigelloides here were previously screened for the presence
were sensitive to all the tea and coffee extracts of insecticides and chemical fertilizers and no
tested. Escherichia coli and Pseudomonas aeru- residues were detected. The heat treatments
ginosa were resistant. inherent in the preparation of tea and coffee
To determine whether the observed inhibition probably preclude the involvement of proteins
of growth by these extracts was bactericidal or in this effect. Tea has been reported to have
bacteriostatic, one strain of each of Staph. antiviral properties (John & Mukundan 1979;
aureus, V . parahaemolyticus and V . cholerae 0-1 Green 1949) which appear to be due to tannin.
was tested using the viable cell method. Figure 1 Preliminary experiments in our laboratory
shows that all the tea and coffee extracts killed suggest that tannic acid does indeed inhibit the
V. parahaemolyticus within 3 h. Growth inhibi- growth of bacteria.
tion by pu-erh tea had not been observed for
this organism with the agar diffusion method We thank Yasuo Kudoh for the generous gift of
(Table 1). In the case of Staph. aureus, black and the strains used in this study and George M.
pu-erh tea showed stronger bactericidal activity Fukui for reviewing this manuscript. This work
than green tea or coffee. Vibrio cholerae 0-1 was was supported by grants from the Japanese
killed within 1 h by all four extracts (data not Cholera Panel of the United States-Japan Co-
shown). operative Medical Science Program.
Bactericidal tea and coflee 125
a b



' 50

Time ( h 1
Fig. 1. Bactericidal effects of tea and coffee against a, Staphylococcus aureus or b, Vibrio parahaemolyticus. 0,
black tea; 0 ,green tea; x , pu-erh tea; A, coffee. No. of colonies of control, Staph. aureus, V . parahaemolyticus:
0 h, 430,205; 1 h, 456,229; 3 h, 595,363; 24 h, 155 x lo6, 10 x lo6, respectively.

GREEN, R.H.1949 Inhibition of multiplication of influ- tea, caffeine and tannic acid. Indian Journal of
enza virus by extracts of tea. Proceedings of the Medical Research 69,542-545.
Society for Experimental Biology and Medicine 71, MITSUHASHI, S. et al. 1981 Determination of minimum
84-85. inhibitory concentration (MIC). Chemotherapy 29,
JOHN, T.J. & MUKUNDAN, P. 1979 Virus inhibition by 76-79.