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This oral presentation examines the effects of dietary chitosan, extracted from the carapace of freshwater crab Sartorianaspinigera, on body weight, liver weight, and food intake of hypercholesterolemic albino rats. The study explores the potential of chitosan as a natural remedy for obesity, without the side effects associated with surgery and synthetic drugs.
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ORAL PRESENTATION FORGLOBAL OBESITY MEETING,2018 Effect of Dietary Chitosan extracted from carapace of freshwater crab Sartorianaspinigera on body weight ,liver weight and food intake of hypercholesterolemic albino rats. Presented by SHINY E.C. KACHHAP RESEARCH SCHOLAR IN ZOOLOGY ASSISTANT PROFESSOR RANCHI UNIVERSITY,INDIA
Statement of Problem What is obesity? Obesity is a medical condition in which excess body fat has accumulated to the extent that it may have an adverse effect on health.It is defined by body mass index(BMI) . If a person’s body weight is at least 20% higher than it should be,then the person is considered obese. A body BMI more than 30 is considered to be an indicator of obesity. Obesity has been linked to a number of health complications such as : Type II diabetes, Hypertension, Hypercholesterolemia, Fatty liver disease, Stroke, Infertility, and Breathing problems.
Some Data.. According to a survey conducted in 2015,about 600 million adults and 100 million children were found to be obese in 195 countries due to high fat food consumption in uncontrolled manner. In America According to Centers for disease control and Prevention(CDC),(2015-16),it has been found that 93.3 million American adults and 13.7 American children and teens are clinically obese. In India 10 million cases are noted every year India has the second highest number of obese children in the world after China. In Ezypt Ezypt tops the list for adult obesity at about 35 percent.
To cure obesity.. Bariatic surgery :It decreases intestinal absorption of food by bypassing part of digestive system.It has shown to cause various side effects such as diarrhoea, anemia Liposuction :it removes fat cells but shows side effects of heart disease and diabetes. Synthetic drugs: Phenylpropanolamine(PPA)is an appetite suppressent but shows an increased risk of haemorrhagic stroke in females. Sibutramine is also related to cardiovascular diseases. Orlistat is also used to loose weight but also has side effects.
Requirement of natural remedy Due to all the above side effects caused by costly surgeries and synthetic medicines,the society is moving towards the treatment by natural origin . Such medicines are not only cost effective but are also biocompatible and do not show side effects. Zootherapeutic medicines formed by crustacea,fungi,etc.,are highly recommended.
SARTORIANA SPINIGERA • It is commonly called as Rusty crab. • It is a freshwater edible crab abundantly found in Jharkhand’s shallow water bodies such as rice fields,canals,ponds. • It comes under LC(least concern)category in the IUCN red list. • Sartoriana spinigera is a delicacy and its soup is medically used by Jharkhand’s tribal people during cough and cold, eye and gastrointestinal infections,to attain faster recovery and strength after a severe health issue and also to women after delivery. CLASSIFICATION: Phylum: Crustacea Class:Malacostraca Order:Decapoda Family:Gecarcinucidae
CHITOSAN From the carapace of Sartorianaspinigera ,a fibre ,Chitosan is extracted . Fig: structural formula of Chitosan Chitosan is a linear polysaccharide formed by deacetylation of chitin.Chitosan is composed of two units: 80% deacetylated unit-ß 1-4 linked D glucosamine 20%acetylated unit-ß 1-4,N-acetyl-D-glucosamine Chitosan is a cation and thus gets attracted to anions such as bile acids and thus does not allows them to get absorbed in the intestine. Some reports suggest that chitosan extracted from marine crabs,shrimps, have the property of controlling cholesterol level in the body,and causing satiety.But no such work has been performed on the chitosan extracted from freshwater crab Sartoriana spinigera.
Specimen of Sartoriana spinigera Chitosan extracted from carapace of S.spinigera Chitosan fed to hypercholesterolemic albino rats Weight of rats measured on 1st and 30th day of experiment Food intake of rats were measured Liver weight of rats were measured on 30th day
Hypercholesterolemic rat Weight of rats measured on 1st and 30th day of experiment Liver weight of rats were measured on 30th day Food intake of rats were measured
Fig:Carapace of S.spinigera Fig:Foaming of carapace powder after addition of 2N HCl during demineralization Fig:Brown coloured carapace powder Fig:Cream coloured chitosan obtained
Characterization of Chitosan 1.Degree of deacetylation: Average degree of deacetylation of chitosan=78.53±2.03% 2.Characterstics of chitosan obtained from freshwater crab Sartorianaspinigera:
ANIMALS AND DIET 20 albino rats ,each measuring about 80 grams were purchased from local rearing laboratory. Rats were maintained at room temperature.There were 12 hours daytime light between 6AM to 6PM and 12 hours of dark Rats were fed with respective diets at 9 AM and 9PM.Water and basal diet were provided ad-libitum for 3 days as acclimatization period. Experimental diets were given for 30 days. Group A-basal diet Group B-high fat diet Group C-high fat diet + 5%chitosan Group D-high fat diet+ 5% hypolipidemic drug(ezetimibe)
Analytical method Estimation of body weight gain: Body weight of rats of all groups were measured on 1st(initial) and 30th(final) day to calculate the body weight gain. Estimation of food intake : Food intake of rats of all groups were calculated on 1st,3rd,5th,7th,14th,21st and 30th day to study the satiety in rats. Estimation of liver weight: On the final day of experiment,rats were sacrificed to obtain their liver,that were weighed to study the increase or decrease in weight of liver. Estimation of lipid content in liver: Lipid content of liver was calculated by drying the obtained liver and measuring the lipid by folch method Estimation of fecal excretion in rats: Total fecal matter was measured on1st,3rd,5th,7th,14th,21st and 30th day and were weighed to study the comparitive excretion of all groups. Also total lipid excreted by the rats were calculated by folch method.
Statistical analysis All values were expressed as the mean of 5 animals. The difference obtained in analysis of food intake of different groups on different experimental days were analysed by ANOVA. .Difference between body weight gain of group A and B and that of B and C group and that of C and D group were statistically analysed by Student’s t test to test the significance. Difference between average of food intake of group A and B and that of B and C group and that of C and D group were statistically analysed by Student’s t test to test the significance. Difference between liver weight of rats of group A and B and that of group B and C and that of group C and D group were statistically analysed by Student’s t test to test the significance.
Body weight gain * **=significant at 0.1% * *=significant at 1%
Graph 1: representing body weight gain in 30 days of 4 groups fed with respective diets • Group A showed least body weight gain. • Group B showed highest body weight gain. • Group C showed body weight gain lesser than that of group B and group D • Group D showed body weight gain lesser than group B.
Graph 2: representing average amount of food consumed by rats of different groups The result shows that group fed with chitosan consumed least amount of food amongst all groups.
Liver weight * * *=significant at 0.1% level * * *=significant at 0.1% level * * =significant at 1% level
Graph 3: representing wet weight of liver of four groups fed with respective diets on 30th day of experiment • Group A showed least weight of liver • Group B showed highest weight of liver • Group C showed liver weight lower than group B and very near to group A • Group D showed liver weight lower than group B and same as group C.
Graph 4: representing lipid content of liver(%) of four groups fed with respective diets on 30th day of experiment
Fecal excretion of lipid Group C fed with chitosan excreted highest amount of lipid .
Graph 5: representing fecal excretion of lipid of four groups fed with respective diets on 30th day of experiment
According to R.Ylitalo et al (2002),cationic Chitosan binds with the anionic lipid and fatty acid chains and does not allows them to get absorbed in the intestine, rather makes them get excreted out of the body and thus body weight is decreased Intestinal wall Cationic chitosan Anionic fatty acids Excreted out of the body
It has been observed that Chitosan fibre,when enters the digestive tract ,it swells and causes the feeling of fullness in stomach thereby causing satiety Digestive tract Swelled chitosan in acidic ph of stomach
According to Pan,H. (2016),Liver contains LDL receptors that are responsible for clearance of LDL from circulation. Chitosan increases LDL receptor mRNA expression so that more LDL gets removed from liver liver LDL receptors in liver that attract LDL and clear them from liver
Chitosan leads to an increase in Leptin hormone ,which causes satiety in the stomach thereby stimulating the individual to consume lesser food Increased concentration of Leptin i.e., the satiety hormone Increased concentration of chitosan in body Individual consumes lesser amount of food
CONCLUSION We have also conducted pathological studies on histology and transmission electron microscopy(TEM) of liver of rats of all four groups.In liver of rats fed with high fat diet and no chitosan,large amount of lipid dropolets were found.Whereas,inchitosan fed rats,lipid droplet concentration was very low. Total cholesterol level,triglyceride level and LDL level have also been found to be lesser in rats fed with chitosan.All these evidences indicate that chitosan has hypocholesterolemic activity and antiobesity activity.
Chitosan,a biopolymer extracted from Freshwater crab Sartoriana spinigera has antiobesity properties as it causes satiety in the gut . • Chitosan eliminates fat from the body and thereby inhibits its deposition causing a balanced body and liver weight. • Chitosan is a biocompatible,biodegradable and non-toxic substance that can be encouraged as an antiobesity medicine rather than synthetic drugs.