Top Banner
The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9
22

The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Jan 03, 2016

Download

Documents

Adam Black
Welcome message from author
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
Page 1: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

The Effects of Vitamins B6 and C on Yeast Growth

By: Gianmarco FrezzaPittsburgh Central Catholic HS

Grade 9

Page 2: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Rationale

• Humans need vitamins B6 & C from their diets or as supplements

• Vitamin B6 RDA: 1.3 mg/day

• Vitamin C RDA males: 90 mg/day, females: 75 mg/day

• Does vitamin B6 or C affect the microbial flora of the body?

Page 3: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Microbial Flora

• Refers to microorganisms living in or on the body (gastrointestinal tract)

• Little is known about the association between humans and their flora

• Effects are mutualistic, parasitic, pathogenic, and commensal

• Perform functions beneficial to the host, including the manufacture of essential vitamins, and the prevention of colonization by undesirable microbes

• Human foods and supplements may have unintended effects on the flora populations and their functions

Page 4: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Saccharomyces cerevisiae

• A fungi• One of the most

intensively studied eukaryotic model organisms

• Commonly known as yeast

• Can be used as a model for human microflora

Page 5: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Vitamin B6 (Pyridoxine)

• Important for more than 100 enzymatic reactions involved in metabolism and brain development during pregnancy and infancy

• Helps the immune system function properly

Page 6: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Vitamin C (ascorbic acid)

• An antioxidant, protects cells from damage from free radicals

• Helps the immune system function properly• Growth and repair of tissues• Protects against cardiovascular disease• Prevents scurvy

Page 7: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Purpose

• To test if Vitamin B6 or C have an effect on S. cerevisiae growth individually or synergistically

Page 8: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Hypothesis

• Null: the vitamins WILL NOT have an significant effect on yeast growth individually or synergistically.

• Alternate: the vitamins WILL have a significant effect on yeast growth both individually and synergistically.

Page 9: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Materials• 36 sidearm flasks• Aluminum foil• Lab coat• Disposable rubber gloves• Micropipetters• Sterile micropipette tips• Sterile dilution fluid

(100mMKH2PO4, 100mMK2HPO4, 10mMMgSO4, 1mMNaCl)

• Macropippetter• Sterile macropipette tips• 3 Incubators

•1% stock solution vitamin B6•1% stock solution vitamin C•YEP media (1% yeast extract, 2% peptone, 2% glucose)•S. Cerevisiae•Klett Spectrophotometer

Page 10: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Procedure

1. 7.5 mL of YEP media was pipetted into each sterile sidearm flask.

2. Four replicates of the following combinations of variables were prepared as follows.

3. 0.5 mL of S. cerevisiae (KU=100) was pipetted into each flask.4. The flasks were covered with a sterilized aluminum foil cap.5. The 36 sidearm flasks were placed in incubators (32⁰C), and

readings were taken every half hour for six hours in a Klett spectrophotometer.

Page 11: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Mixtures by GroupsS.Cerevisiae(mL)

YEP media(mL)

Concentrations (B6, C) %

Vitamin C (mL)

Vitamin B6 (mL)

SDF (mL)

Group 1 0.5 7.5 [0%] B6 [0%] C 0 0 2

Group 2 0.5 7.5 [0%] B6 [0.01%] C

0.1 0 1.9

Group 3 0.5 7.5 [0%] B6 [0.01%] C

1 0 1

Group 4 0.5 7.5 [0.01%] B6 [0%] C

0 0.1 1.9

Group 5 0.5 7.5 [0.01%] B6 [0.01%] C

0.1 0.1 1.8

Group 6 0.5 7.5 [0.01] B6 [0.1] C 1 0.1 0.9

Group 7 0.5 7.5 [0.1%] B6 [0%] C 0 1 1

Group 8 0.5 7.5 [0.1] B6 [0.01] C 0.1 1 0.9

Group 9 0.5 7.5 [0.1%] B6 [0.1] C 1 1 0

Page 12: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Concentrations of the Vitamins within Groups

[0%] (0) B6 [0.01%] (L) B6

[0.1%] (H) B6

[0%] (0) C 00 L0 H0

[0.01%] (L) C 0L LL HL

[0.1%] (H) C 0H LH HH

Page 13: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Vitamin Effects on Yeast Growth

60 120 180 240 300 3600

20

40

60

80

100

120

140

Group 1 CGroup 2 0L Group 3 0HGroup 4 L0Group 5 LLGroup 6 LHGroup 7 H0Group 8 HLGroup 9 HH

Group 1 CGroup 2 0L Group 3 0HGroup 4 L0Group 5 LLGroup 6 LHGroup 7 H0Group 8 HLGroup 9 HH

Time (min.)

KU

L= [0.01%] H= [0.1%]B, C

Page 14: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Groups at 180 minutes

0% C 0.01% C 0.1% C0

20

40

60

80

100

120

0% B0.01% B0.1% B

Concentrations of vitamin C

Klett Units

Two factor ANOVA synergy P= 0.1162

Single Factor vitamin B P= 0.958538

Single factor vitamin C P= 0.972004

Page 15: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Groups at 360 minutes

0% C 0.01% C 0.1% C0

20

40

60

80

100

120

140

0% B0.01% B0.1% B

Concentrations of Vitamin C

Klett units

Two factor ANOVA synergy P= 0.1785

Single Factor vitamin B6 P= 0.000429

Single factor vitamin C P= 0.16963

Page 16: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Conclusion• The null hypothesis is accepted for all trials other than vitamin B6 at 360 minutes exposure.

• Vitamin B6 and C did not have a significant effect on yeast growth individually or synergistically.

Page 17: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Limitations and Extensions

Limitations• Only two

concentrations of the variables used

• Only growth was tested

• Six hour duration

Extensions•More concentrations

•Longer Duration

•Study survivorship

•More species

Page 18: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Resources• http://en.wikipedia.org/wiki/VitaminC

• www.ods.od.nih.gov/factsheets/VitaminC-HealthProfessional/

• http://en.wikipedia.org/wiki/VitaminB6

• www.ods.od.nih.gov/factsheets/VitaminB6-HealthProfessional/

• http://en.wikipedia.org/wiki/Saccharomyces_cerevisiae

• Mark Krotec, PTEI

• www.epa.gov/epawaste/nonhoz/municipal/

Page 19: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Raw Data at 180 minutes

Groups (Replicant 1) Klett units

Group 1 71

Group 2 73

Group 3 87

Group 4 73

Group 5 59

Group 6 41

Group 7 62

Group 8 79

Group 9 101

Groups (Replicant 2) Klett units

Group 1 70

Group 2 84

Group 3 80

Group 4 72

Group 5 78

Group 6 35

Group 7 57

Group 8 94

Group 9 100

Page 20: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

Raw Data at 360 minutesGroups (Replicant 1)

Klett units

Group 1 128

Group 2 107

Group 3 127

Group 4 92

Group 5 105

Group 6 24

Group 7 118

Group 8 105

Group 9 113

Groups (Replicant 2) Klett units

Group 1 125

Group 2 108

Group 3 120

Group 4 94

Group 5 98

Group 6 25

Group 7 119

Group 8 111

Group 9 117

Page 21: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

ANOVA at 180 minutes

Page 22: The Effects of Vitamins B6 and C on Yeast Growth By: Gianmarco Frezza Pittsburgh Central Catholic HS Grade 9.

ANOVA at 360 minutes