Antimicrobial Effect of Cranberry Powder on E.coli ATCC 25922 in Apple Cider and Ground Beef ...

122 Pages·2010·1.65 MB·English

Antimicrobial Effect of Cranberry Powder on E.coli ATCC 25922
in Apple Cider and Ground Beef
by
Jane Palakeel
A Research Paper
Submitted in Partial Fulfillment of the
Requirements for the
Master of Science Degree
With a Major
ill
Food & Nutritional Sciences
Approved: 6 Semester Credits
The Graduate School
University of Wisconsin-Stout
August, 2010
2
The Graduate School
University of Wisconsin-Stout
Menomonie, WI
Author: Palakeel, Jane J.
Title: Antimicrobial Effect of Cranberry Powder on E.coli ATCC
25922 in Apple Cider and Ground Beef
Graduate DegreelMajor: MS Food and Nutritional Sciences
Research Adviser: Cynthia Rohrer, PhD
MonthNear: August, 2010
Number of Pages: 122
th
Style Manual Used: American Psychological Association, 6 edition
Abstract
The possibility of using cranberry powder as an antimicrobial agent was studied by
examining its effect on the growth of E coli A TCC 25922 in apple cider and ground beef and its
organoleptic properties in beef patties. The standard plate counts (SPC) results showed that
cranberry powder (2% w/w) reduced Ecoli ATCC 25922, by 0.43 log CFU/mL, in apple cider
(p>0.05) and 0.08 log CFU/g in ground beef (p<0.05) compared to their respective controls on day
6. Thus demonstrating that 2% w/w cranberry powder posses antimicrobial benefits when
incorporated in ground beef but not as well in apple cider. RT-PCR showed results contrary to that
observed in SPC. Cranberry powder (2% w/w) significantly reduced E coli ATCC 25922 (p<0.05),
by 0.83 log CFU/mL in apple cider but showed less antimicrobial activity in ground beef compared
to the control. At least one hundred panelists evaluated beef patties supplemented with and without
cranbeny powder. No significant difference for color, cooked beef flavor, cardboard flavor, grainy
3
texture, aftertaste, overall acceptance and preference were found among beef patties with 0%, and
2% w/w cranberry powder on day 0 (p>0.05) except for sweetness and juiciness which were both
significantly greater in cranberry supplemented beef patties. Therefore, 2% w/w concentration of
cranberry powder can be considered an ideal concentration to be added into ground beef patties.
4
The Graduate School
University of Wisconsin-Stout
Menomonie, WI
Acknowledgements
I would like to express my sincere gratitude to the following individuals for their help,
contribution, effort, encouragement in completion of my thesis work. I would like to thank Dr.
Cynthia Rohrer, for her tremendous contribution to my thesis completion by involving in framing,
editing the research proposal, designing the sensory tests, editing my thesis, research poster and for
her constant encouragement. Dr. Kitrina Carlson for helping set up the microbiology experiments,
developing the methodology, providing work space and supplies, and for great moral support and
help in completing my thesis work. I would like to thank Dr. Carolyn Barnhart for editing my
thesis and instilling the zeal in me to complete my thesis work. I would like to thank Dr. Marcia
Miller for providing me valuable inputs for my research and for editing my thesis. I would like to
specially thank the Biology department, for providing the laboratory equipment, reagents and
microbiology media and the space to carry out the experiments for the research. I would like to
thank the Cranberry Seed Oil Company for providing the cranberry powder to conduct my research.
I would like also thank Dr. Steve Nold for guiding, helping me with data analysis and directing me
towards completing the last components of my research work. I would like to thank my family
members and friends, without their valuable support, advice and help I would not have been
successful in achieving my goal. Last but not least, I would like to thank God for giving me the
strength to overcome all the uphill battles I faced during my research work.
5
Table of Contents
............................................................................................................. , .... Page
Abstract ............................................................................................................ 2
List of Tables ...................................................................................................... 9
List of Figures .................................................................................................... 10
Chapter 1: Introduction ................... ·. ...................................................................... 12
Statement of the Problem .............................................................................. 15
Research 0 bj ecti ves .................................................................................... 15
Assumptions of the Study ............................................................................. 16
Definition of Terms .................................................................................... 16
Limitations of the Study ................ '" .. '" ....................................................... 17
Chapter II: Literature Review .................................................................................... 18
History of Cranberries ............................... '. ............................................ , ..... 18
Cranberry Species and Attributes ..................................................................... 19
Cranberry Cultivation Factors ......................................................................... 19
Dry Harvesting ................................................................................. 20
Wet Harvesting .................................................................................2 0
Processing of CranbelTies ..............................................................................2 1
Physiochemical Quality ................................................................................. 21
Polyphenols ..................................................................................... 22
Phenolic Acid Chemistry of Cranberries ................................................... 22
Flavonoid Chemistry of Cranberries ......................................................... 23
Flavonols ....................................................................................... 24
6
Flavanols ....................................................................................... 26
Proanthocyanidins ............................................................................ 27
Anthocyanins ................................................................................... 28
Resveratrol ...................................................................................... 29
Nutritive Quality ........................................................................................ 30
Health Benefits ......................................................................................... 31
Urinary Tract. ................................................................................. 31
Helicobacter Pylori ........................................................................... 33
Prevention of Kidney Stone Information .................................................. 34
Cardiovascular Diseases ..................................................................... 35
Antimicro bial Properties ...................................................................... 36
Antioxidants .................................................................................... 39
Anticancer. ..................................................................................... 40
Dental Plaque .................................................................................. 41
Cranberry Powder. ..................................................................................... 42
Engineering of Cranberry Powder ......................................................... .42
Antimicrobial Effects of Phenolic and Organic Acids ............................................ .43
Inhibition of E.coli 0157 :H7 in Apple Cider. ...................................................... .44
Inhibition ofE.coli 0157: H7 in Ground Beef.. .................................................... 46
Real Time Polymerase Chain Reaction ............................................................. .47
Sensory Evaluation ..................................................................................... 48
Chapter III: Methodology ....................................................................................... 51
Sample Selection and Description .................................................................... 51
7
Bacterial Strain ................................................................................. 52
Reagents ........................................................................................ 52
Preparation of Mueller Hinton Agar. ....................................................... 53
Methanol Extraction and Vacuum Filtration ....................................................... 53
Kirby Bauer Disk Diffusion (Antimicrobial Disk Method) ...................................... 53
Microanalysis for Apple cider. ....................................................................... 54
Apple Cider Preparation ..................................................................... 54
pH Measurement ............. " .............. , ............. , ..... , ............................ 56
Microanalysis for Ground Beef. ..................................................................... 56
Ground Beef Preparation ..................................................................... 56
pH Measurement .............................................................................. 57
Sensory Evaluation .................................................................................... 57
Genomic DNA Extraction ............................................................................ 59
Real Time PCR Analysis .............................................................................. 60
Data Analysis ........................................................................................... 61
Chapter IV: Results and Discussion .......................................................................... 62
Methanol Extraction ................................................................................... 62
Antimicrobial Effect of Cranbeny Powder In Apple Cider. ..................................... 64
Antimicrobial Effect of CranbelTY Powder In Ground Beef. .................................... 69
Comparison of E. coli Population in Cranbeny Powder Added Apple Cider and Ground
Beef. ..................................................................................................... 76
Quantification by RT-PCR .................................................................................7 7
Sensory Evaluation .................................................................................... 87
Chapter V: Conclusion ......................................................................................... 92
8
Recommendations ..................................................................................... 96
References ....................................................................................................... 97
Appendix A: Sensory Advertisement. ................. '" ................................................. 121
Appendix B: Beef Patty Scorecard .......................................................................... 122
9
List of Tables
Table 1: Apple Cider and Ground Beef Treatments .................................................... 55
Table 2: Samples and Three Digit Codes Assigned ................................................... 59
Table 3: Diameter of Zone ofInhibition of CranbelTy-Methanol Extract on
Saureus ATCC 25923, Ecoli ATCC 25922 and Styphimurium .................................. 64
Table 4: Colony Forming Units (CFU/mL) of Ecoli ATCC 25922 in Apple Cider with and
without Cranbeny Powder at different Dilutions on Day 0, Day 2, Day 4, and Day
6 ................................................................................................... 66
Table 5: Means and Confidence Intervals of Colony Fonning Units (log CFU/mL) in Apple
Cider with and without Cranberry Powder .................................................. 67
Table 6: Colony Forming Units (CFU/g) of Ecoli ATCC 25922 in Ground Beef with and
without Cranberry Powder at different Dilutions on Day 0, Day 2, Day 4, and Day
6 ................................... ~ ............................................................... 71
Table 7: Means and Confidence Intervals of Colony Forming Units (log CFU/g) in Ground Beef
with and without Cranbeny Powder ......................................................... 72
Table 8: Concentrations of Serially Diluted Ecoli DNA with adding Cranbeny Powder.. ....... 81
Table 9: Concentrations of Serially Diluted Ecoli DNA without adding Cranbeny Powder... 81
Table 10: Mean Concentrations of Ecoli ATCC 25922 DNA in Apple Cider with
and without Cranbeny Powder. .............................................................. 82
Table 11: Mean Concentrations of Ecoli ATCC 25922 DNA in Ground Beef with
and without Cranbeny Powder .............................................................. 84
Table 12: Mean Sensory Attributes of Control and Cranbeny Powder added-Ground Beef
Patties .................. , ............................. .......................................................... 87
10
List of Figures
Figure 1: Caffeic acid, Sinapic acid, p-Coumaric acid and Benzoic acid ........................... 23
Figure 2: Quercetin and Myricetin ........................................................................ 26
Figure 3: Procyanidin dimer AI and Procyanidin dimer A2 ........................................... 28
Figure 4: Cyanidin and Peonidine ........................................................................ 30
Figure 5: Apple Cider ( concentrate) and Ground Beef Treatments .................................... 52
Figure 6: Serial dilution and spread plating technique ................................................. 57
Figure 7: Dilution of sample and stomaching in stomacher ........................................... 58
Figure 8: Sample Preparation and taste testing in sensory booth ..................................... 60
Figure 9: C1000 Thermal Cycler .......................................................................... 61
Figure 10: Inhibition of Ecoli ATCC 25922 in Apple Cider with and without CranbelTY Powder
on Day 0, Day 2, Day 4, and Day 6 .............................................................. 69
Figure 11: Plate Counts of E coli A TCC 25922 in Ground Beef with Cranberry Powder on Day
2 .......................................................................................................... 74
Figure 12. Plate Counts of Ecoli ATCC 25922 in Ground Beef without CranbelTY Powder on
Day 2 ............................................................................................. 74
Figure 13: Inhibition of E coli ATCC 25922 in Ground Beef with and without Cranberry Powder on
Day 0, Day 2, Day 4 and Day 6 .................................................................................. 75
Figure 14: Inhibition of Ecoli ATCC 25922 in both Apple Cider and Ground Beef having added
CranbelTY Powder on Day 0, Day 2, Day 4 and Day 6 ...................................... 77
Figure 15: Melting Curve for Ecoli ATCC DNA sample at 10-1 dilution ............................ 78
Figure 16. Amplification of Serially Diluted E coli DNA with and without Cranberry
Po\vder. ......................................................................................... 79