BRIEF REPORT

F o l i a Microbiol. 29, 3 4 8 - - 352 (1984)

A Comparative Study of Raw Garlic Extract

and Tetracycline on Caecal Microflora and
Serum Proteins of Albino Rats
]~.N.

SttASH, KANTH, S.C. BASAPPA

and V.

SRE:ENIVASA MURTHY

Central Food Technological Research Institute, M y s o r e -- 570 013, I n d i a

Received September 6, 1983

A B S T B A C T . S t u d i e s c o n d u c t e d o n t h e c o m p a r a t i v e a c t i o n of r a w garlic e x t r a c t a n d t e t r a c y c l i n e
h y d r o c h l o r i d e o n e q u a l c o n c e n t r a t i o n s s h o w e d t h e r a w garlic e x t r a c t to be a m o r e p o t e n t a n t i m i c r o b i a l a g e n t t h a n t e t r a c y c l i n e . T h e r e v e r s i o n to n o r m a l c y of caecal microflora a n d s e r u m
proteins after the withdrawal of test materials indicated an inverse relation between intestina.
microflora a n d s e r u m globulins.

The broad antimicrobial effectiveness of raw garlic extract is a well estabhshed fact (Subramanyan et al. 1958; Prasad and Sharma 1980). The broad
antibacterial nature of tetracyclines is universally recognized and tetracyclines are extensively used in therapy. However, intestinal bacteria are
known to develop resistance to some of the antibiotics on prolonged usage
(Drasar and Hill 1974). Under the circumstances, it was envisaged to compare
the action of raw garlic extract and tetracycline on equal concentrations on
caecal microfl0ra and serum proteins.
Garlic bulbs were purchased from the local market and tetracycline hydrochloride capsule were procured from a dispensary. Antibiotic assay employing
S. aureus culture was carried out following the procedure of Collins (1964) to
determine the concentration of tetracycline hydrochloride equivalent to
0.5 g / m L of raw garlic extract, and was fixed to be 4 m g / m L of tetracycline
hydrochloride.
Eighteen adult male Wistar strain albino rats of similar weight range were
allotted into groups of three animals each. They were fed with a control corn
starch diet ad libitum (Shashikanth 1983). Two groups were maintained as
controls, one for 3 d, the other for 6 d. The remaining four groups were t r e a t e d
as experimental. The ~third and fourth group rats were administered with
2 m L of 0.5 g / m L of fresh raw garlic extract per animal per d a y for a period
of 3 d successively. Similarly, fifth and sixth group rats were administered
with 2 m L of a 4 m g / m L of tetracycline hydrochloride in water for 3 d.
At the end of 3 d the administrations were withdrawn and animals belonging to first, third and fifth groups, representing control, raw garlic and

1984

C O M P A R I S O N OF G A R L I C E X T R A C T A N D T E T R A C Y C L I N E

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K.N. S H A S H I K A N T H et al.

Vol. 29

Fro. 1. Serum protein profile of control rats (gels scanned

in Beckman Spectrophotometer gel scanning model 26);
A albumin, P prealbumin, ~l, ~2, ~l, ~2 and y globulins.

tetracycline groups, respectively, were sacrificed, blood was collected a n d

stored under refrigerated conditions. The caecum was taken out aseptically
and the caecal contents were analyzed for total streptococci, coliforms,
lactobacilli, aerobes and anaerobes employing Streptococcus faecalis agar, eos
in methylene blue agar, t o m a t o juice agar, total plate count agar and Brewer's
anaerobic agar respectively, after suitable serial dilutions to get 3 0 - - 3 0 0
colonies per plate. The serial dilutions for plating anaerobes were carried
out employing anaerobic mineral solution (Bryant and Robinson 1961). All
the plates were incubated at 37 i 1 ~ for 1 d. The anaerobic plates were
incubated at 37 ~ 1 ~ for 3 d in an anaerobic incubator (National Appliance
Company, Portland, Oregon, USA) filled finally with 90 ~/o of pure Iolar
grade I Nitrogen and 10 % of carbon dioxide, after prior recyclings with
carbon dioxide alone three times. ,Triplicates were maintained and the counts
were subjected to statistical analysis using the _F-test.
Polyacrylamide gel electrophoresis (PAGE) of the sera was done following
the procedure of Davis (1964). The gels were scanned in a B e c k m a n Spectrophotometer Scanning Model 26.
The results obtained are shown in Table I. At equal concentrations b o t h
tetracycline and raw garlic extract showed pronounced inhibition as compared to the controls. Nearly a hundred-fold decrease was observed in t o t a l
streptococci and coliforms followed b y ten-fold decrease for total lactobacilli, aerobes and anaerobes. After 3-d successive administrations of
both the test materials caecal microflora counts for tetracycline administered
rats had significantly lowered, as compared to those of garlic and controls,
especially in cases of total streptococci, aerobes and anaerobes. Though the

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COMPARISON OF GARLIC EXTRACT AND TETRACYCLINE

35t

FIG. 2. S e r u m p r o t e i n profile o f r a w garlic fed r a t s ; left : r a w garlic a d m i n i s t e r e d for 3 d; right: r a w garlic a d m i n i s t e r e d for 3 d A- 3 d c o n t r o l diet;
for s y m b o l s see Fig. 1.

caeeal flora of rats fed raw garlic extract had significantly lower counts they
were significantly more than those of tetracycline fed rats.
However, on feeding only control diet after withdrawal of the test materials, the microflora of the caeca of tetracycline fed animals tended to revert
to normalcy faster than those after garlic, indicating a prolonged inhibitory
action of garlic extract on intestinal microbes.
Bacteria develop faster resistance to commercially available antibiotics
(Drasar and Hill 1974). At certain times prolonged administrations of antibiotics m a y lead to the colonization of yeasts and moulds which are hazardous
to the hosts. On the other hand, natural antimierobial agents such as garlic
can inhibit both bacteria and moulds apart from its hypoglyeaemic and
hypeeholesteremic actions.
The serum protein profile of control, raw-garlic extract, and tetracycline
fed animals are presented in Figs. 1 -- 3. After the withdrawal of the administrations and feeding of only control diet it was observed t h a t the serum
protein profile, especially globulins tended to r e v e r t to normalcy faster in
tetracycline-fed animals than those fed with garlic, which clearly demonstrated the higher inhibitory activity of garlic than tetracycline.
The data also suggested t h a t there existed correlation between the intestinal
microflora and serum globulins. This was clearly seen in the fact t h a t serum
globulins decreased after the administration of garlic and tetracycline and
reverted to normalcy after their withdrawal. This rallies with the findings
of BhaVanlshankar (1981) and Berg and Savage (1979) who found an inverse
correlation between the intestinal laetobaeilli count and globulin levels in the

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K . N . S H A S H I K A N T H etal.

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F~o. 3. Serum protein profile of tetracycline fed rats; left: tetracycline

adzninistered for 3 d; right: tetracycline a d m i n i s t e r e d for 3 d H- 3 d control
diet; for s y m b o l s see Fig. 1.

blood. This was confirmed b y a comparative s t u d y of feeding raw and boiled

garlic extracts for a period of four weeks to albino rats (Shashikanth 1983).
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influence on bile acid metabolism. PhD Thesis, U n i v e r s i t y of Mysore 1980.
BERO R.D., SAVAGE D.C.: I m m u n o l o g i c a l responses a n d microorganisms indigenous to the
gastrointestinal tract. Amer.J.Clin.Nutr. Qt, 1364 (1972).
BRYANT M.P., ROBINSON I.M.: Some nutritional r e q u i r e m e n t s of t h e genus Ruminococcus. Appl.
Microbiol. 9, 96 (1961).
COLLINS C.H.: Microbiological Methods, p. 296. ~Butterworths, L o n d o n 1964.
DAvis J3.J.: D i s c oleetrophoresis, i . Method a n d application to h t t m a n s e r u m proteins. Ann.
N.Y.Acad.Sci. 1 2 1 , 4 0 4 (1964).
DRASAR B.S., HILL M.J.: Human Intestinal Flora, 1st Ed. Academic Press, L o n d o n 1974.
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SU~RAF[]Iff.ANYAN V., SREENIVASA MURTHY V., KRISHNA MURTHY K., SWAMINATHA~ M.: The
effect of incorporation of garlic in t h e diet on t h e intestinal mieroflora of rats. Ann.Biochem.
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