The staining response of a certain bacteria depends on its cell wall components
(http://www.ruf.rice.edu/~bioslabs/bios318/staining.htm)
Overview
Overview Mordant
Tanic acid, Iodine, etc., Forms a mordant-dye-protein complex which can not be decolorized in gram positive bacteria but is extracted by alcohol in gram negative bacteria
(Histopathologic techniques by Gregorios, M.D)
Overview Mordant
In Gram stain: It is added after the crystal violet and before the alcohol in the gram reaction.
Tannin
Repels many organisms that cause wood decay Used to preserve leather
(Nabors, 2005)
Tannic Acid
Commercial form of Tannin Weak acid Has mordant properties
Main Problem
Is the Tamarind (Tamarindus indica) Seed Extract effective as a Mordant in Gram Staining?
Sub Problems
What are the physical and chemical properties of the seed husk extract in comparison with the Grams Iodine? Will the physical and chemical properties of the tannic acid from the seed husk extract level or exceed those of the Grams Iodine?
Sub Problems
Using spectrophotometry, what is the concentration of the tannic acid from the seeds husk extract? Will the concentration of the tannic acid from the seed husk extract level or exceed those of the Grams Iodine concentration?
General Objective
To prove the effectiveness of Tamarind (Tamarindus indica) Seed Extract as Mordant in Gram Staining.
Specific Objectives
To be able to use tamarind leaves extract as a mordant in Grams staining To compare the effectiveness of tamarind leaves extract as a mordant in Grams staining to Grams iodine To determine the best solvent concentration to be used in Tamarind extract. To identify the concentration of tannic acid that will be effective to be used as mordant in Grams staining.
Significance
For the scientific community It can be a stepping stone in the evolution of the use of the local resources for medical studies. It can also give an idea to medical professions to come up with a new way or reagents of staining.
Significance
For the agricultural community The success of this study will be noted as advancement not only in the scientific studies but also for the agricultural growth of our country. Tamarind is one of the abundant crops in the Philippines and through this study, more opportunities will be developed for the use of more Tamarind products.
Significance
For the general public The use of the organic raw material will be promoted which can be in replacement of the chemically produced Grams Iodine.
Significance
For the medical technology field The medical technologist will be able to find an alternative for Grams iodine which can be a cause of many problems in their health. It can be the start of finding some alternative reagents for the toxic ones used in the laboratory for the benefit of the medical technologist.
Research Design
Experimental (True Exerimental)
Methodology
I. Collection II. Extraction ii.1. Aqueous: water ii.2. Basic/ Alkaline: dioxane ii.3. Acidic: acetic acid ii.4. Alcoholic: ethanol
Methodology
III. Measurement of Effectiveness III.i. Collection of Bacteria
E. coli and S. Aureus from ATCC
Nutrition broth
Methodology
III.ii. Test tube Preparation
Methodology
IV. Calculations v.1. Absorbance v.2. Relative color strength to the nearest 0.1%
Calculations
Beer Lamberts law: A = log10(1/T ) (1)
A is the absorbance at wavelength of minimum transmittance T is the transmittance (in decimal fraction form, where 100% = 1.0) at wavelength of minimum transmittance.
Calculations
Relative color strength to the nearest 0.1% Fs = 100 (A2C1/A1C2)
Fs = 100 (A2/A1)
Fs is relative color strength in percent C is the dye concentration subscript 1 refers to the reference dye subscript 2 refers to the dye under test Assuming C1 = C2, the following simplified equation is obtained:
Gantt Chart
OBJECTIVES 1. Topic proposal 2. Title proposal 3. Approval of proposals 4.RRL 5. Chapter 1 draft 6. Revision of chapter 1 and RRL 7. Chapter 3 draft 8. Revision of chapter 3 9. Preparation for experimentation 10. Final draft of chapters 1, 2, and 3. JUNE JULY AUG.
SEPT
OCT
Gantt Chart
OBJECTIVES 11. Experimentation 12. Analysis and interpretation of data 13. Chapters 4 and 5 draft 14. Revisions of chapter 4 and 5 15. Final manuscript 16. Oral defense NOV OCT DEC JAN FEB