Bioresearch Bulletin (2010) 2: 91-93
ORIGINAL COPY
Impact of Clinical Pathogens on the Gut Toxicity of Selected Albino Mice
ABHINAV CHAURASIA1, ABHISHEK CHAUDHARY1, R.BALAJI RAJA2*
Received: 02 August 2010 /Accepted: 22 August 2010 /Published online: 13 September 2010
© Bioindica Press 2010
ABSTRACT
The impact of various clinical pathogens on the
gut toxicity has remained an elusive question as the re-
sults have been highly inconclusive till date. The mice
were classified as normal and test with difference in their
feeding cycle. Various clinical pathogens used were Sal-
monella typhi, Shigella and Escherichia coli. The intro-
duction of the clinical pathogens was followed by the
removal of various visceral organs including small intes-
tine, liver, spleen and kidney. Microbiological analysis of
these organs using agar media, employing standard
streak method yielded both qualitative and quantitative
details about the toxicity caused by clinical pathogens
used in these albino mice. Small intestine was found to
having the highest accumulation of the pathogens in
comparison with other visceral organs with 89 x 10 -3
CFU. A clear correlation was found between the gut tox-
icity and the nature of the clinical pathogen employed.
The study can be extended to the mechanism beneath the
differences arising in the various visceral organs when
encountered with clinical pathogens.
Keywords: Salmonella typhi, Shigella, Escherichia coli,
Toxicity, Visceral organs
INTRODUCTION
Toxicity induced in the various visceral organs
by the hyper presence of clinical pathogens has been a
long standing common phenomenon (Brandtzaeg 1988,
Kerr 1990). Various kinds of toxicity are associated with
pathogenic organisms, gut toxicity, hepatotoxicity,
nephrotoxicity, neurotoxicity etc… The pathogenic mi-
crobial organisms which form the primary cause for most
of these toxicities are bacteria followed by virus, yeast
and other types (Brandtzaeg and Sollid 1987). Gut toxic-
ity seems to be the most common toxicity induced in
Abhinav Chaurasia1 Abhishek Chaudhary1
Department of Biotechnology, SRM University,
Kattankulathur – 603203, Tamilnadu, India
R.Balaji Raja2*
Department of Biotechnology,
National Institute of Technology, Raipur, India.
Telephone: +91-09942574378,
email: balajiraja_vlr@yahoo.com
91
humans and animals by these pathogenic bacteria. The
toxicity inducement of the bacteria seems to be following
a different mechanism altogether when it comes to the
organs being affected (Cancellieri and Fara 1985). Re-
ceptor bound mechanism becomes the case in gut toxic-
ity whereas intermediate compound formation is the
mechanism in case of nephrotoxicity. The case of neuro-
toxicity is the most complicated in terms of mechanism
as it involves a ‘stop-go’ approach where the toxicity is
not induced in one step but in a phased wise manner
(Carter and Collins. 1974).
Pathogenic microbes also differ in their toxicity
mechanism based on the species which is involved. For
example Salmonella species follows a molecular ap-
proach whereas if Shigella species is involved, it has
‘stop-go’ approach (Chau and Tsang 1981). There is a
distinct difference in the quantification of infection be-
tween the genus and the species as well. The same genus
with a different species yields a different result altogether
when it comes to level of toxicity.
The organs which are most affected by any class
of microbes are the visceral organs. Visceral organs
mainly include small intestine, large intestine, spleen,
kidney and the liver. Each organ shows its own charac-
teristic resistance against the advent of infection induced
by the pathogenic microorganisms (Childers and Bruce
1989, Kantele 1986). Liver is the most potent in putting
up a stiff resistance against a wide spectrum of patho-
genic bacteria and it is the most successful in warding off
the potential threat posed by the bacteria. The above
mentioned is the reason why liver is the least affected
when it comes to clinical pathogen infection. In compari-
son the gut seems to be the most prone organ when it
comes to clinical infection by the microbes.
As gut is already containing a good number of
microbes which assist in digestion, the mechanism to
identify the harmful bacteria becomes week resulting in
it being the soft target as far as clinical infestation is con-
cerned (Edelman 1986, Hohmann 1978). The mechanism
being the major factor does not stop the mention about
the compositional difference in these organs. Liver is
made of thicker tissues as it is the primary organ for me-
tabolism and detoxification. Spleen is made up of com-
paratively less hard tissues which are in transition phase
between being hard and soft. Kidney is composed of
spongy tissues which are easy target for the invading
microbes and small intestine is the easiest target because
of the tissue composition (Finlay 1988, Gorbach 1989).
Bioresearch Bulletin (2010) 2: 91-93
Small intestine is made up of the softest tissues when
compared with other organs in question.
MATERIALS AND METHODS
Pre-Clinical Standardization
12 male albino mice were obtained from TANU-
VAS (Tamil Nadu Veterinary and Animal Sciences Uni-
versity), Madhavaram, Chennai. They were 6 weeks old
and weighed around 25-30 grams. The mice were placed
in 2 cages labeled as Control (C) and Test (T). They were
left for acclimatization period of 15 days in their respec-
tive cages. The mice were fed once a day with standard
rodent chow and provided with distilled water. All ex-
periments were performed under controlled conditions
(temperature [21 +_ 2°C], humidity, and a 12-h light-
dark cycle).
Infection
Salmonella typhi (MTCC Number-237), Shigella
(MTCC Number-121), Escherichia coli (MTCC Number
-095) were procured from IMTECH, Chandigarh and
were sub cultured in a specific medium (LB Agar me-
dium). 2 test mice were infected with 0.1 ml of 10 -2 E.
coli , 2 were infected with Salmonella typhi and 2 were
infected with Shigella sp. on the 16th day. On the 17th
day 0.1 ml of 10-4 E. coli , Salmonella typhi and Shigella
sp were injected through subcutaneous route using gas-
tric tube (Merck Instruments, USA). It was ensured that
during injection all the mice were healthy and did not
have any physical wounds.
Harvesting of Visceral organs
One mouse taken from both groups (Test and
Control), anaesthetised, dissected and the Visceral organs
were harvested (Liver, spleen, small intestine and kid-
ney). The harvested organs were preserved in a formalin
solution.
Calculation of CFU
200 mg of the visceral organ was grinded well
using a mortar and pestle and made in to 5 dilutions, 10 -1
to 10-5. Dilution of 10-3 was taken for the 2 samples
(control and test). The CFU was calculated using the
formula,
CFU = Number of colonies
Dilution factor
RESULTS AND DISCUSSION
As far as the Liver is concerned, Escherichia coli
infection was found to be having minimum impact. The
pathogen count in the control mice was found to be 93 x
10-3 whereas the pathogen count in the case of test mice
was found to be 28 x 10-3. In the kidney, the pathogen
count in the control mice was found to be 60 x 10 -3
whereas the pathogen count in the case of test mice was
found to be 8 x 10-3. Spleen had the pathogen count in
the control mice as 38 x 10-3 whereas the pathogen count
in the case of test mice was found to be 4 x 10 -3. Small
intestine had the maximum pathogen count among the
visceral organs with control mice having a count of 98 x
92
10-3 whereas the pathogen count in the case of test mice
was found to be 41 x 10-3.
As far as the Liver is concerned, Salmonella ty-
phi infection was found to be having minimum impact.
The pathogen count in the control mice was found to be
81 x 10-3 whereas the pathogen count in the case of test
mice was found to be 23 x 10-3. In the kidney, the patho-
gen count in the control mice was found to be 77 x 10 -3
whereas the pathogen count in the case of test mice was
found to be 14 x 10-3. Spleen had the pathogen count in
the control mice as 38 x 10-3 whereas the pathogen count
in the case of test mice was found to be 10 x 10 -3. Small
intestine had the maximum pathogen count among the
visceral organs with control mice having a count of 90 x
10-3 whereas the pathogen count in the case of test mice
was found to be 32 x 10-3.
As far as the Liver is concerned, Shigella sp.
infection was found to be having minimum impact. The
pathogen count in the control mice was found to be 65 x
10-3 whereas the pathogen count in the case of test mice
was found to be 17 x 10-3. In the kidney, the pathogen
count in the control mice was found to be 78 x 10 -3
whereas the pathogen count in the case of test mice was
found to be 13 x 10-3. Spleen had the pathogen count in
the control mice as 44 x 10-3 whereas the pathogen count
in the case of test mice was found to be 9 x 10 -3. Small
intestine had the maximum pathogen count among the
visceral organs with control mice having a count of 69 x
10-3 whereas the pathogen count in the case of test mice
was found to be 15 x 10-3.
The liver seems to have been least affected by all
the clinical pathogens whereas the small intestine is the
most affected by all three pathogenic bacteria. The im-
pact of these clinical pathogens seems to be affected by
the organisation of the tissues and the mechanism fol-
lowed by the system in which the particular organ is as-
sociated with. Further studies in the same line can solve
many unanswered questions about the mechanism of
every organ when encountered with each clinical patho-
gen.
ACKNOWLEDGEMENTS
We would like to thank the management of SRM
University for allowing us to carry out this work at their
premises. Our sincere thanks go to Dr.Shivashekhar, vice
principal, SRM Medical college who helped with histo-
pathological analysis. The help of Dr. Sekar, veterinary
officer, Animal House, SRM University; Mr. Prab-
hakaran, technician, Animal House, SRM University;
Mr.Jaganath Patra, Technician at Pathology lab, SRM
Hospital, Mr. Jayaprakash, Lab Assistant, Department of
Microbiology, SRM Hospital is really appreciable.
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