EFFECT OF TEMPEH AND SHRIMP

MEAL IN THE TREATMENT OF

ALUMINIUM CHLORIDE INDUCED
ALZHEIMERS IN RATS

Under the Guidence of- PRESENTED BY:

Dr.S.RAMACHANDRAN, Darlanka Yamini Bangaram

M.Pharm., Ph.D. Denngam Phom
Head, Department of Pharmacology Gorrela Likhitha
Penmatsa Nikhitha Sai
Rayapalli Jaya Satyapriya
AIM AND OBJECTIVE

 AIM :

Role of Tempeh along with shrimp in the experimental models of

Alzheimer's disease and oxidative stress .
 OBJECTIVE
The objectives of present study are to evaluate the following parameters :

 Effect of Tempeh , shrimp , and Tempeh and shrimp combination on memory performance in elevated plus maze
paradigm in aluminum chloride treated rats .
 Effect of Tempeh , shrimp and Tempeh + shrimp combination on spatial navigation task in aluminium chloride
treated rats .
 Effect of Tempeh , shrimp and Tempeh + shrimp combination on oxidative stress parameters in aluminium
chloride treated rats .

•a . Lipid peroxidation

•b . Superoxide dismutase catalase

 Effect of Tempeh , shrimp , and Tempeh + shrimp combination on acetylcholine Esteras activity in aluminium
 PLAN OF WORK
  MATERIALS AND METHOD

 MATERIALS AND METHOD

• This work was aimed to investigate the prophylactic and therapeutic role of Tempeh and Indian Shrimp on aluminum chloride
induced Alzheimer’s disease in rats.

• Methods: The test meal will be prepared in required ratio after processing each component by the recommended procedure
and it has to be tested for its Prophylactic role. The male Wistar rats are the animals needed for study and have to be divided
into six groups. Group I must be given with distilled water, Group II has to receive AlCl 3 (100 mg/kg, p.o.), Group III has to

receive Rivastigmine (1 mg/kg, p.o.), Group IV has to receive AlCl 3+ Test meal Dose 1, and Group V has to receive AlCl 3+
Test meal Dose 2 for 60 days. At the end of the study various parameters like behavioral and biochemical investigations have
to be assessed to confirm the success of treatment in Alzheimer’s.
 Preparation of Tempeh & Estimation of Vit B6, B12 & Serotonin

• About 1000g of dehulled soybeans has to be soaked in cold water at

room temperature for 18 h. The soaked soybeans have to be washed
thoroughly and steamed for 40min. After that, the steamed soybeans
have to be cooled down to room temperature and subsequently mixed
with Rhizopus oligocarpous. The inoculated beans have to be packed
into perforated polyethylene plastic bags and incubated for 30h at 30˚C
in incubator. After that it has to be dried, ground, and extracted with
deionized water by cold maceration process. The water extracts of
preparation have to be lyophilized using the freeze dryer. Lastly, the
lyophilized powder has to be stored at 4˚C in an airtight amber
container
  Preparation of shrimp powder and estimation of Selenium:

• Shrimp has to be purchased from rural market of Kakinada, Andhra

Pradesh. They have to be dried in the sun light and grounded into fine
powder. A 1.5% of 2 2,4-DNPH-NEDA reagent mixture has to be
prepared by dissolving 1g of NEDA and 005g of 2,4-DNPH by adding
2 drops of concentrated HCL and volume made up to 100ml with
distilled water.

• 1ml of test shrimp solution is to be transferred in to 10ml flask,

to which 1 drop of HCL and 2ml of 1.5% of 2 2,4-DNPH-NEDA reagent
mixture have to be added. The mixture has to be allowed to stand for
10min with occasional shaking for the completion of the reaction. The
contents were diluted to mark with distilled water and absorbance was
  RESULTS AND DISCUSSION
RESULTS :

 EFFECT OF TEMPEH ( T ) AND SHRIMP ( S ) COMBINATION ON

MEMORY PERFORMANCE IN ELEVATED PLUS MAZE PARADIGM IN
ALUMINUM CHLORIDE TREATED RATS -

• In the elevated plus maze task , control group of animals shows a mean
transfer latency on day 10. With in 90 sec all the rats entered the closed arm
. Aluminum Chloride treated rats showed a significant difference (p <0.005)
of transverse when compared with control group on day10 indicating that
chronic aluminum chloride treatment - induced memory impairment . In
contrast , co - administration of standard Rivastigmine ( p < 0.005 ) and
tempeh + shrimp ( 100 mg / kg + 100mg / kg ; p.o . ) , Tempeh low ( 200mg
/ kg ; p.o ) , Tempeh high ( 400mg / kg ; p.o ) , Shrimp low ( 200 / mg / kg ;
Fig.- Effect Of Tempeh ( T ) And Shrimp ( S ) Combination
p.o ) and Shrimp high ( 400mg / kg ; p.o ) with aluminum chloride On Memory Performance In Elevated Plus Maze Paradigm
( 40.mg/kg;i.p ) show significant decrease ( p < 0.001 ) In Aluminum Chloride Treated Rats
  RESULTS AND DISCUSSION

 EFFECT OF TEMPEH , AND SHRIMP COMBINATION ON

OXIDATIVE STRESS PARAMETERS IN ALUMINIUM CHLORIDE
TREATED RATS .

• Chronic administration of aluminum chloride for 30days showed

significant ( p < 0.005 ) increase in oxidative damage as evidence by
increased MDA and depleted catalase and SOD enzyme activity as
compared to naive group.Concurrent administration of standard and
tempeh + shrimp ( 100 mg / kg + 100mg / kg ; p.o . ) , Tempeh low
Rivastigmine ( p < 0.01 ) ( 200mg / kg ; p.o ) , tempeh high (400mg / kg ;
p.o ) , Shrimp low ( 200 / mg / kg ; p.o ) and Shrimp high (400mg / kg ;
p.o ) with aluminum chloride ( 40.mg/kg;i.p)shows significantly decrease
( p < 0.001 ) potentiated their protective effects (decreased MDA levels
and restored SOD and catalase levels ) as compared to their effects alone
Fig. Effect of tempeh , and shrimp combination on
and control group oxidative stress parameters in aluminium chloride
treated rats
  RESULTS AND DISCUSSION

 EFFECT OF TEMPEH , AND SHRIMP COMBINATION IN HISTOPATHOLOGICAL

INVESTIGATION OF BRAIN SECTION IN DIFFERENT STUDIED GROUPS :

• Microscopic examination of brain section of control rat showed normal morphological

structure of the hippocampus. On the other hand , microscopic investigation for brain
section of aluminum induced rat ( AD - induced group ) demonstrated various sizes of
amyloid plaques in the hippocampus. Histological investigation of brain section of AD -
induced rats treated with Rivastigmine and / or shrimp in a dose of 100mg / kg , 200 mg /
kg revealed more or ss normal histological structure of the hippocampus and all amyloid
plaques that are formed under the influence of AIC13 administration disappeared, as well
as the treatment with Tempeh in a dose of 200mg / kg , 400mg / kg . showed normal
histological structure of the hippocampus , but with few disturbance in hippocampus cells
arrangement. Moreover , the treatment with Tempeh in a dose of 100mg / kg and shrimp in Fig.No.1.12. Effect of tempeh , and shrimp
a dose with 100 mg / kg . showing normal histological structure of hippocampus with combination on acetylcholine esterase activity
in aluminum chloride treated rats
dislocation of some hippocampus cells in AD - induced rats and with few disturbance in
hippocampus cells arrangement associated with dislocation of some cells in AD induced
rats .
  CONCLUSION

 Tempeh contains significant amounts of vitamin B12, isoflavones, folic acid, calcium, protein etc. which may be responsible
in neurological activity. The results of the present study indicate that the Tempeh which is a fermented soya product along
with the shrimp which is a marine food source shows the potentiation role in the treatment of aluminum chloride induced
Alzheimer’s disease not only by neurofibrillary changes, but also from seizures and/or motor impairments. The present
research has opened for further research in the development of potent role of tempeh along with shrimp in Alzheimer’s
disease Vitamin B12 is very rich in non vegetarians diet when compared with vegetarian diet but ,Tempeh which is a
fermented product contain vitamin B12 due to fermentation by using Rhizopus oligocarpous and shrimp which is very cheap
and low cost available marine food source shows significant role in improving neurological conditions like Alzheimer's
disease ,it may also serve as a possible nutritional intervention and may be used as future nutritional supplement to combat
the neurological disorders in humans.
THANKYOU