Bachelor Thesis Presentation on Probiotics

EFFECT OF PROBIOTICS ON HEALTH STATUS OF OBESE

SUBJECTS

BY

Ms. C . D. Prathyusha (06J41A2305) Ms. P. B Kavya (06J41A2309)

B.Tech (4-2) Department of Biotechnology

External Guide Dr.M.Shiva Prakash, Assistant Director

National Institute of Nutrition (ICMR) Hyderabad

Distribution of Dominant Bacteria

The Benefits Of Bacteria

• The word bacteria bears a negative connotation, but in truth, there are both harmful and beneficial bacteria. How often have kids heard their mothers mention bacteria as the reason their hands need to be washed before eating?

• The truth is, a lot of the bacteria found on surfaces are harmful. But there is a plethora of beneficial bacteria contained inside our bodies. These bacteria work to remove toxins and other dangerous elements from our bodies, and promote overall health and wellness.

• Having more of these bacteria in our bodies can be quite beneficial because of the removal of harmful toxins our innate bacteria populations might have missed.

Toxic and Non Toxic Bacteria

Introduction of additional beneficial bacteria into our bodies

Proboitic bacteria then add themselves to the existing bacteria

Promoto better health and can also be used to treat more harmful conditions like autism .

Probiotic Bacteria L.bulgaricus S.thermophilus

PROBIOTICS

“Living Micro organism which when administered in adequate amount confer a health benefit on host.” (FAO 2000 )

Elie Metchnikoff (1907) is considered to be inventor of Probiotics.

Minora Shirota (1930s) suggested that right mix of bacteria in gut could prevent disease

Promotion of health status using Probiotics introduced a new concept of

Probiotic Therapy.

Antibiotics & Probiotics

• While antibiotics are utilized to kill bacteria, probiotics, on the other hand, are bacteria in themselves. We often think of bacteria as being exclusively detrimental but many of them are necessary and beneficial, such as PROBIOTICS.

• These particular bacteria are essential in maintaining the natural balance of organisms in the intestines and ensuring proper digestion.

• Human body consists of 85% harmful bacteria and 15%beneficial bacteria

• Probiotics are designed to replace beneficial bacteria in the body.  Probiotics are ingested like a standard capsule pill, and the bacteria are then released into the body.  Taking probiotics as a supplement is an excellent idea for all people who don’t receive proper nutrition.

• Because probiotics are all natural, there’s no need to worry about foreign chemicals or toxins entering the body and causing damage.  When foreign chemicals or toxins are the culprit, probiotics is the solution

beneficial

harmful

Suppression of exogenous pathogens

Suppression of endogenous pathogens

Reduction of risk of diseases

Immune enhancement

Functions of Probiotics

• Inclusion of Probiotic bacteria in fermented dairy product enhances their value as better therapeutic functional food.

• Curd and yoghurt are important fermented foods• Organism used as Probiotic in fermented food items include : Lactobacillus bulgaricus, L. casie, L. rhamnoses, L.

acidophillus, Streptococcus thermophilus, Bifidobacterium etc

Probitics in Fermented Food

OBESITY

Obesity is an excess of body fat to such an extent as to have an adverse effect on well being and health (WHO ).

Health problems related to Obesity Include :

• Diabetes.• Heart Disease.• Increased cholesterol.• Lower immune status.• Lower Nutritional levels leading to ill health.

To study the effect of Probiotics supplementation ( Indian curds) on Immune status by analyzing immunoenzyme marker Adenosine deaminase in obese subjects

To observe the changes in the micronutrient levels (Zn, Ca & P) before and after Probiotics supplementation

To assess the effect on lipid profile status (Total Cholesterol ,Triglyceride and HDL cholesterol) after supplementation of Probiotic in obese subjects.

OBJECTIVES

METHODOLOGY

Different Bio chemical Reactions

Yogurt Preparation

ESTIMATION OF CALCIUM • Calcium in alkaline medium combines with Ortho-

cresolpthalein complex one to form peuple coloured complex. Intensity of the colour formed is directly proportional to the amount of calcium present in the sample

• Reaction: calcium+OCPC purple colored complex

• Calculation: Calcium in mg/dl = Abs. T / Abs. S × 10

RESULTS OF CALCIUMCategory Number of

subjects‘0th’ day ‘7th’ day

Obese individuals (supplemented group)

10 8.62 ± 4.34 7.38 ± 1.63

Controls (Non supplemented group)

6 6.25 ± 2.5 10.62 ± 8.26

Normal range: 8.7-11.0 mg/dl

Estimation of Adenosine Deaminase • Adenosine Deaminase hydrolyses adenosine to ammonia and

inosine. The ammonia formed further reacts with phenol and hypochlorite in an alkaline medium to form a blue indophenol complex with sodium nitroprusside acting as a catalyst. Intensity of the blue coloured indophenol complex formed is directly proportional to the amount of ADA present in the sample.

• REACTION: Adenosine + H2O Ammonia + Inosine Ammonia + Phenol + Hypocholrite Blue Indophenol Complex

Alkaline medium • CALCULATIONS:

Total ADA activtyU/L: Abs.T-AbsSB / Abs.S-Abs.B × 100

RESULTS FOR ADA Category Number of

subjects‘0th’ day ‘7th’ day

Obese individuals (supplemented group)

10 23.32±13.42 16.98±5.41

Controls (Non supplemented group)

6 23.02±4.44 20.7±2.65

Normal range: 15-30 U/L

ESTIMATION OF TOTAL CHOLESTEROL• Cholesterol esters are hydrolyzed to produce cholesterol.

Hydrogen peroxide is then produced from oxidation of cholesterol by cholesterol oxidase.

• The indicator quinonemine is formed from hydrogen peroxide and 4-aminoantipyrine in the presence of phenol and peroxide.

• The absorption of the red quinoneimine dye is proportional to the concentration of cholesterol in the sample.

• Calculation: Cholesterol in mg /dl= Abs. of (T)/Abs. of (S) x 200mg/dl

RESULTS OF TCCategory Number of

subjects ‘0th’ day ‘7th’ day

Obese individuals (supplemented group)

10 161.24±33.87 160.97±23.65

Controls (Non supplemented group)

6 152.0 ±8.0 218 ± 1 18.0

Normal ranges: 140-250 mg/dl

Estimation of HDL Cholesterol • Low density lipoproteins (LDL and HDL) and

chylomicron fractions are precipitated by precipitating reagent .

• After centrifugation, the cholesterol concentration in the HDL(high density lipoprotien) fraction remains in the supernatant in this phase and is determined by an enzymatic method.

• Calculation: HDL cholesterol = AC/AS x50 mg/dl x 2

Results of HDL Cholesterol

Category Number of subjects

‘0th’ day ‘7th’ day

Obese individuals(supplemented group)

10 29.67 ±8.20 40.70 ±7.75

Controls ( non-supplemented group)

6 31.74 ±4.94 34.84 ±6.95

Normal range: 30-75 mg/dl

Estimation of Triglycerides • Triglycerides are determined after enzymatic

hydrolysis with lipases. The quinonemine indicator is formed from hydrogen peroxide, 4-aminophenazone, and chlorophenol under the catalytic influences of peroxidases.

• Calculation: Triglycerides = AT/AS x 200 mg/dl

Results of Triglycerides Category Number of

subjects ‘0th’ day ‘7th’ day

Obese individuals (supplemented group)

10 133.59 ± 94.85 140.01 ± 41.76

Controls (non- supplemented group)

6 179.46 ± 8.89 190.16 ± 23.33

Normal range: 40-165 mg/dl

Thank You