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A Study on the Potential Mutagenicity of Overcooked Hardboiled Eggs using Allium cepa test Russel Christine B. Corcino Germaine D. Arda

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Background of the Study Eggs Major part of the daily diet of humans Can be cooked in different ways (ex. Boiling) People may sometimes overcook eggs Generates hydrogen sulfide (H 2 S) harmful to the body Coagulates the structure of the egg People still consume overcooked eggs

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Statement of the Problems 1.How long is the normal cooking time of an egg to be considered? 2.Is there a difference in the physical characteristics of an overcooked hardboiled egg and a normal cooked hardboiled egg? 3.Is there a significant difference on the mitotic index among the overcooked hardboiled eggs and normal cooked hardboiled egg? 4.Is there a significant difference between the mitotic indexes of overcooked eggs cooked in different time intervals?

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Scope and Limitations Boiling is the method used in cooking the eggs Three time intervals (including the normal cooking time) Positive and negative controls for comparison Allium cepa test is used (microscopic analysis only) No chemical analysis made Study is conducted at MSU-IIT IDS Chemistry Laboratory II

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General Procedure

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TREATMENTDESCRIPTION ACooked at normal cooking time BCooked 2 minutes beyond normal cooking time CCooked 5 minutes beyond normal cooking time SAMPLEDESCRIPTION AExposed to Treatment A BExposed to Treatment B CExposed to Treatment C Positive Controlexposed to 0.05% colchicine Negative Controlexposed to distilled water Table 1. Treatments Done to the Hardboiled Egg Table 2. List of the Different Sample of Roots

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Allium cepa test 1.Distilled water 2.Treatment solution (45 minutes) exposure to the treatment 3.Distilled water 4.Fixative solution (30 minutes) freeze the cells 5.Distilled water 6.1 M HCl and left (10-12 minutes) to soften the root tip 7.Distilled water.

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Results of the Experiment A.Normal Cooking time: 15 minutes B.Physical Characteristics (Overcooked vs. Normal) (a)Normal-cooked egg, (b) Egg cooked 2 mins. beyond normal, (c) Egg cooked 5 mins. beyond normal

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Fig. 1. Mitotic Indices of the Onion Roots Exposed to the Different Treatments C. Microscopic Analysis

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Can be caused by CARCINOGENICITY, which is a type of MUTAGENICITY

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Fig 2. Normal Mitosis of Allium cepa roots (from the left: Interphase, Prophase, Metaphase, Anaphase, Telophase) (Huss, 2007)

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Fig. 3. Chromosomal aberrations detected in Replicate 2 exposed to treatment B: (A) chromosomal bridging during early anaphase, (B) chromosomal stickiness.

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Hypothesis Testing (One-way ANOVA) H o : μ 1 = μ 2 = μ 3 = μ 4 = μ 5 H 1 : at least one differ from the others α = 0.05 (level of significance) Table 3. Basis for Groups for ANOVA and Post hoc Analysis Group #Treatment 1Positive Control 2Negative Control 3Treatment A 4Treatment B 5Treatment C

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Reject Ho since p-value=0.0003 is less than α = 0.05. Proceed to Post hoc Analysis! Table 4. ANOVA table SourceSSdfMSF p-value Treatment2,385.2618404596.315460015.19.0003 Error392.4813361039.2481336 Total2,777.74317614

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Table 5: Post hoc Analysis: p-values for pairwise t-tests Group # Group 2Group 3Group 1Group 5Group 4 Mean 31.8913535.3543144.6988459.7860163.19930 Group 231.89135 Group 335.35431.5138 Group 144.69884.0312.0977 Group 559.78601.0003.0007.0145 Group 463.19930.0001.0003.0047.5197

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Treatment A vs. Controls Group 3 has no significant difference with group 2 since their p-value is equal to 0.5138 which is greater than α= 0.05. Group 3 has no significant difference with group 1 since their p-value is equal to 0.0977 which is greater than α= 0.05. μ 1 = μ 3 μ 2 = μ 3

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Treatment B vs. Controls Group 4 has a significant difference with group 2 since their p-value is equal to 0.0001 which is less than α= 0.05. Group 4 has a significant difference with group 1 since their p-value is equal to 0.0047 which is less than α= 0.05. μ 1 μ 4 μ2 μ4μ2 μ4

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Treatment C vs. Controls Group 5 has a significant difference with group 2 since their p-value is equal to 0.0003 which is less than α= 0.05. Group 5 has a significant difference with group 1 since their p-value is equal to 0.0145 which is less than α= 0.05. μ 1 μ 5 μ2 μ5μ2 μ5

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Treatment A vs. Treatment B and Treatment C Group 3 has a significant difference with group 4 since their p-value= 0.0003 is less than α= 0.05. Group 3 also has a significant difference with group 5 since their p-value= 0.0007 is less than α= 0.05. μ3 μ4μ3 μ4 μ 3 μ 5

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Treatment B vs. Treatment C Group 4 has no significant difference with group 5 since their p-value is equal to 0.5197 which is greater than α= 0.05. μ 4 = μ 5

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Summarizing the Results μ positive = μ A μ negative = μ A μ positive μ B μ negative μ B μ positive μ C μ negative μ C μ A μ B μ A μ C μ B = μ C

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Conclusions 1.The normal cooking time to be considered for hardboiled eggs is 15 minutes. 2.There is a difference on the physical characteristics among the normal cooked hardboiled eggs and overcooked hardboiled eggs. 3.There is a significant difference between the mitotic indices in the roots of the Allium cepa exposed to normal cooked hardboiled egg and overcooked hardboiled egg. 4.There is no significant difference between the mitotic indices in the roots of the Allium cepa exposed to overcooked egg cooked at different time intervals.

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Recommendations The researchers recommend the ff: To do more study about this research using other mutagenicity tests like Drosophila, etc. To test the possible effects of raw eggs to the cells. To test other variety of egg samples such as ducks, birds and native chicken.

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THE END

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