Effectiveness of sodium bicarbonate mixed with saba banana (Musa acuminate x balbisiana) and red banana (Musa acuminate ‘Red Dacca’) sap as an alternative mordant in gram stain /
Ronnel M. Mojica.
- Indang, Cavite : Cavite State University- Main Campus, 2018.
- xiv, 126 pages: illustrations ; 28 cm.
Research Study (Bachelor of Science in Medical Technology) Cavite State University.
Includes bibliographical references.
College of Nursing (CON) College of Nursing (CON)
EFFECTIVENESS OF SODIUM BICARBONATE MIXED WITH SABA BANANA (Musa acuminata x balbisiana) AND RED BANANA (Muse acuminata Red Dacca') SAP AS AN ALTERNATIVE MORDANT IN GRAM STAIN Research Study RONNEL M. MOJICA CAMILLE ANNE B, RODILA PATRISHIA MAE O, TORRES College of Nursing CAVITE STATE UNIVERSITY Indang, Cavite May 2018 EFFECTIVENESS OF SODIUM BICARBONATE MIXED WITH SABA BANANA (Musa acuminata × balbisiana) AND RED BANANA (Musa acuminata 'Red Dacca') SAP AS AN ALTERNATIVE MORDANT IN GRAM STAIN Undergraduate Research Study Submitted to the Faculty of Department of Medical Technology College of Nursing Cavite State University Indang, Cavite In partial fulfillment of the requirements for the Research MEDT 24B 00017891 Effectiveness of sodium bicarbonate mixed with saba banana (Musa acuminate x 648.5 M72 2018 RS-831 MOJICA, RONNEL M. RODILA, CAMILLE ANNE B. TORRES, PATRISHIA MAE O. May 2018 ABSTRACT MOJICA, RONNEL, RODILA, CAMILLE ANNE, TORRES, PATRISHIA MAE, Effectiveness of Sodium Bicarbonate Mixed with Saba Banana (Musa acuminata × balbisiana) and Red Banana (Musa acuminata 'Red Dacca') Sap as an Alternative Mordant in Gram Stain. Undergraduate Research, Bachelor of Science in Medical Technology. Cavite State University. Indang, Cavite. May 2018. Adviser: Dr. Hosea dL. Matel. This study aimed to (a) determine the effectiveness of Sodium bicarbonate mixed with Saba Banana (Musa acuminata × balbisiana) and Red Banana (Musa acuminata 'Red Dacca') sap can be used as an alternative mordant for gram staining based on color, clarity and stability;(b) determine the effectiveness of sodium bicarbonate mixed withSaba Banana (Musa acuminata × balbisiana) and Red Banana (Musa acuminata 'Red Dacca') sap against Gram's Iodine in microscopic examination. Two trials, eight treatments, three replicates and a control were used in the study, eight bacteria were allocated to each treatment. The stained specimen was observed for its color, clarity and stability. The staining ability of every treatment were developed and categorized. Sodium bicarbonate mixed with Saba Banana (Musa acuminata × balbisiana) and Red Banana (Musa acuminata 'Red Dacca') sap were used as an alternative mordant to Gram's Iodine, exhibited a good color, clarity and stability. The treatment which has the best staining ability has a pH of >8 because of its alkalinity. All concentrations of Sodium bicarbonate mixed with Saba banana (Musa acuminata × balbisiana) and Red Banana (Musa acuminata 'Red Dacca') sap were highly V significant if it will be compared to the staining ability of Gram's Iodine. Saba Banana Sap (Musa acuminata x balbisiana) has the highest rating compared to Red banana in terms of mordant color, clarity and stability. The control and treatments exhibit slightly alkaline pH. Eight bacteria was stained using Gram's Iodine as the control and was also stained using Sodium bicarbonate mixed with Saba Banana (Musa acuminata × balbisiana) and Red Banana (Musa acuminata 'Red Dacca') sap which also demonstrate its effectiveness as compared to Gram's Iodine. vi TITLE PAGE... TABLE OF CONTENTS Page i BIOGRAPHICAL DATA..... ACKNOWLEDGEMENT. ABSTRACT............. TABLE OF CONTENTS... LIST OF FIGURES.............. LIST OF APPENDIX FIGURES. LIST OF APPENDIX TABLES.. INTRODUTION.. Objectives of the Study....... Significance of the Study...................... Time and Place of the Study... Scope and Limitation of the Study.......... Definition of Terms.... Conceptual Framework. REVIEW OF RELATED LITERATURE. Sodium Bicarbonate... Banana... Characteristics of Banana... pH of Banana. Saba Banana.. Characteristics of Saba Banana. ii 111 V vii X xi xiii 1 3 3 5 5 6 6 8 8 9 10 10 10 11 vii Health Benefits of Saba Banana... Red Banana.. 11 12 Benefits of Red Banana... 14 Health Benefits of Red Banana to the Body's Health... 14 Banana Sap... 15 Banana Sap in Medicine. 15 Adverse Effects of Banana... 16 Banana Sap Dye in Fabrics...... Work done on Banana pseudostem..... Banana Sap's Important Constituents.. Storage... 17 17 18 21 Gram Staining... 21 Gram Positive and Gram Negative.... 23 Biosafety Level 1 Bacteria......... 24 List of Gram Positive and Gram Negative Bacteria... 25 Gram Stain Reagents.... 25 Composition of Grams Iodine...... 27 Conversion of gram positive cells to gram negative......... 27 McFarland Standard... METHODOLOGY... Research Design... Preparation of Materials..... 27 29 30 30 Collection of Plant Material.. 30 Extraction of Plant Material... 31 Treatment Preparation.. 33 viii Preparation of Bacteria for Staining...... Smearing Procedure.... Staining Procedure... Determination of Staining Properties... Determination of Stability and Validity... Criteria for Grading... Data Gathering...... Likert Scale... Proper Disposal of Specimen... Statistical Analysis...... RESULTS AND DISCUSSIONS.. SUMMARY, CONCLUSION AND RECOMMENDATION.. Summary... Conclusion. Recommendations....... REFERENCES... APPENDICES.. ix 33 33 34 35 35 37 38 39 40 40 41 67 67 70 72 70 2 2 2 76 LIST OF FIGURES Figure Page 1 Conceptual Framework of the Study....... 2 Illustration of the Grading System... 3 Flowchart of Methodology.. 4 Schematic diagram of the study.. 5 Gram's staining procedure.. 6 Process of preparing the banana sap. 7 Collection and Processing of Plant Material. 8 Extraction and Processing of Banana Sap.... 7 39 78 79 80 81 82 83 9 Treatment Preparation...... 84 10 Bacterial Preparation McFarland Standard.. 85 11 Bacterial Smearing Procedure.. 86 12 Gram Staining Procedure..... 87 13 Decontamination Procedures.. 88 X LIST OF APPENDIX FIGURES Figure 1 Flowchart of methodology Page 78 2 Schematic diagram of the study 3 Gram staining procedure 4 Process of preparing the banana sap 5 Collection and Processing of Plant Material 6 Extraction and Processing of Banana Sap 7 Treatment Preparation 8 Bacterial Preparation McFarland Standard 9 Bacterial Smearing Procedure 79 80 81 82 83 84 85 86 10 Gram Staining Procedure.. 87 88 11 Decontamination Procedures 12 Staphylococcus aureus using Sodium Bicabonate mixed with Saba Banana Sap 89 13 Staphylococcus aureus using Sodium Bicarbonate mixed with Red Banana Sap. 90 14 Staphylococcus epidermidis using Sodium Bicarbonate mixed with Saba Banana Sap 91 15 Staphylococcus epidermidis using Sodium Bicarbonate mixed with Red Banana Sap. 92 16 Bacillus subtilis using Sodium Bicarbonate mixed with Saba Banana Sap 93 17 Bacillus subtilis using Sodium Bicarbonate mixed with Red Banana Sap xi 94 18 Bacillus coagulans using Sodium Bicarbonate mixed with Saba Banana Sap 19 Bacillus coagulans using Sodium Bicarbonate mixed with Red Banana Sap 95 96 20 Enterobacter aerogenes using Sodium Bicarbonate mixed with Saba Banana Sap 97 21 Enterobacter aerogenes using Sodium Bicarbonate mixed with Red Sap 98 22 Escherichia coli using Sodium Bicarbonate mixed with Saba Banana Sap 99 23 Echerichia coli using Sodium Bicarbonate mixed with Red Banana Sap 100 24 Serratia marcescens using Sodium Bicarbonate mixed with Saba Banana Sap.. 101 25 Serratia marcescens using Sodium Bicarbonate mixed with Red Banana Sap 102 26 Pseudomonas aeruginosa using Sodium Bicarbonate mixed with Saba Banana Sap 103 27 Pseudomonas aeruginosa using Sodium Bicarbonate mixed with Red Banana Sap 104 Xii LIST OF APPENDIX TABLES Table 1 Mean scores for color in tests of Between-Subjects Effects for Staphylococcus aureus... 2 Staphylococcus aureus mean scores for color... 3 Mean scores for clarity in tests of Between-Subjects Effects for Staphylococcus aureus...... 4 Staphylococcus aureus mean scores for clarity... Page 105 105 106 106 5 Mean scores for color in tests of Between-Subjects Effects for Staphylococcus epidermidis... 107 6 Staphylococcus epidermidis mean scores for color... 107 7 Mean scores for clarity in tests of Between-Subjects Effects for Staphylococcus epidermidis......... 108 8 Staphylococcus epidermidis mean scores for clarity... 108 9 Mean scores for color in tests of Between-Subjects Effects for Bacillus subtilis... 109 10 Bacillus subtilis mean scores for color.... 109 11 Mean scores for clarity in tests of Between-Subjects Effects for Bacillus subtilis... 110 12 Bacillus subtilis mean scores for clarity... 110 13 Mean scores for color in tests of Between-Subjects Effects for Bacillus coagulans... 111 14 Bacillus coagulans mean scores for color... 111 15 Mean scores for clarity in tests of Between-Subjects Effects for Bacillus coagulans.......... 112 16 Bacillus coagulans mean scores for clarity.. 112 xiii 17 Mean scores for color in tests of Between-Subject Effects for Enterobacter aerogenes... 18 Enterobacter aerogenes mean scores for color. 19 Mean scores for clarity in tests of Between-Subjects Effects for Enterobacter aerogenes... 20 Enterobacter aerogenes mean scores for clarity... 21 Mean scores for color in tests of Between-Subjects Effects for Escherichia coli... 113 113 114 114 115 22 Escherichia coli mean scores for color... 115 23 Mean scores for clarity in tests of Between-Subjects Effects for Escherichia coli... 116 24 Escherichia colimean scores for clarity......... 116 25 Mean scores for color in tests of Between-Subjects Effects for Serratia marcescens... 117 26 Serratia marcescens mean scores for color.. 117 27 Mean scores for clarity in tests of Between-Subjects Effects for Serratia marcescens... 118 118 28 Serratia marcescens mean scores for clarity...... 29 Mean scores for color in tests of Between-Subjects Effects for Pseudomonas aeruginosa... 30 Pseudomonas aeruginosamean scores for color.. 31 Mean scores for clarity in tests of Between-Subjects Effects for Pseudomonas aeruginosa... 32 Pseudomonas aeruginosamean scores for clarity.. xiv 119 119 120 120 EFFECTIVENESS OF SODIUM BICARBONATE MIXED WITH SABA BANANA (Musa acuminata × balbisiana) AND RED BANANA (Musa acuminata 'Red Dacca') SAP AS AN ALTERNATIVE MORDANT IN GRAM STAIN MOJICA, RONNEL M RODILA, CAMILLE ANNE B. TORRES, PATRISHIA MAE O. An undergraduate research study submitted to the faculty of the Department of Medical Technology, College of Nursing, Cavite State University, Indang, Cavite in partial fulfillment of the requirements for the degree of Bachelor of Science in Medical Technology with Contribution No. Prepared under the supervision of Hosea dL. Matel, PhD. and Charmaine V. Rosales, RMT. INTRODUCTION Sodium bicarbonate is a white, crystalline powder that is commonly used as a pH buffering agent, an electrolyte replenisher and systemic alkalizer. It has a pH of 8-9, non-toxic and bitter taste. In microbiology, it is used as an assistant to mordant stain. It may be added just before use to intensify the uptake of color. Banana sap is a well-known extract that can cause stain which cannot be easily removed with bleach and soap. It is extremely sticky which can be used as an alternative adhesive. Sap can be obtained from different parts of the banana tree. It has some special properties relating to various phenomena such as browning of fruits after harvesting caused by its polyphenol oxidase constituent and also permanent staining of