BIO 243 HFCC Chapter 14 Microbes in the Produce Aisle Lab i have the lab manual; the answers r in there once u accept the bid I will provide it Name ______

BIO 243 HFCC Chapter 14 Microbes in the Produce Aisle Lab i have the lab manual; the answers r in there once u accept the bid I will provide it Name ____________________________
Lab 2-6 Fluid Thioglycollate Broth
Read the lab protocol found on pages 95-100 of Microbiology Laboratory Theory and Application by Leboffe and Pierce.
It’s important that you read the lab and data sheets to understand the scope of the lab activity.
Watch ​Basics of Fluid Thioglycollate Media​.
1. Use this space to take notes on the concept of “aerotolerance”. ​Provide a definition, description of the concept​,
and any other information you feel is relevant.
2. Use colored pencils/pens/crayons to color code your notes in this chart, and to complete your sketch of where you
would find each organism type in the FTB test tube at right. Some zones will overlap.
3. Perform a search of the 3 organisms listed on pg 96 in the Materials section. Based on your
research, predict which category these organisms belong to by writing the species name in
the “Aerotolerance” box that they belong in (see example)..
4. Then, draw an arrow next to the test tube at right that points from areas of lowest oxygen
concentration and points to areas with the highest oxygen concentration.
Aerotolerance
Obligate Aerobe
Obligate
Anaerobe
Microaerophile
Facultative
Anaerobe
Ex: Escherichia
coli
Aerotolerant
Anaerobe
Define the term
Predict where you expect to find
this kind of organism in the FTB
Name ____________________________
5. Use this space to identify the properties of the fluid thioglycollate broth. List the various components of the broth and
their purpose, except for resazurin which is listed in #6.
a. How is this broth useful to microbiologists? Why would a microbiologist perform this assay?
6. Purpose of resazurin dye.
a. What is the purpose of resazurin dye in the broth?
b. Where is the resazurin dye located in the broth?
c.
Why is the broth pink at the surface?
7. Why is it important to prepare this broth fresh? Why can’t you store it for days before using it?
8. Suppose you forgot to prepare fresh broth before performing this assay and used an old broth.
a. Which type of organism would be most ​negatively affected​ by old broth?
b. Which type of organism would be most ​positively affected​ by old broth?
Name ____________________________
Lab 2-7 Anaerobic Jar
Read the lab protocol found on pages 101-104 of Microbiology Laboratory Theory and Application by Leboffe and Pierce.
It’s important that you read the lab and data sheets to understand the scope of the lab activity.
Watch ​How to Prepare an Anaerobic Jar
1. In the boxes below, make predictions for which of the three species you expect to see growing in the aerobic plate
and in the anaerobic plate that is incubated in the sealed jar.
Aerobic Plate
Anaerobic Plate in Jar
2. After incubation, you examine the jar before opening it and see that the methylene blue strip is blue (or the
resazurin strip is pink).
a. Based on this, what would you guess the internal environment to be- aerobic or anaerobic?
b. Based on this, how would you expect the growth on the plates inside the jar to differ from the plates
incubated outside the jar?
c.
Based on what you learned in Lab 2-6, predict which of the three organisms would be most affected by
the conditions described above.
3. An alternative to the anaerobic jar is a candle jar, in which a candle is placed and lit with the lid closed to enable
the flame to use the available oxygen. Typically in this system, not all of the oxygen is used. Which types of
organisms would most likely benefit from this environment?
Name ____________________________
Lab 2-8 The Effect of Temperature on Microbial Growth
Read the lab protocol found on pages 105-110 of Microbiology Laboratory Theory and Application by Leboffe and Pierce.
It’s important that you read the lab and data sheets to understand the scope of the lab activity.
Review ​The Effects of Temperature on Microbial Growth
1. Review Figure 2.40. Identify and define the three “cardinal temperatures” that describe where a species can live.
a.
b.
c.
2. Review Figure 2.41
a. At which temperature do thermophiles have the highest growth rate?
b. Are psychrophiles able to survive at 30℃?
c.
What temperature range do psychrophiles and psychrotrophs overlap in?
d. Interpret what the graph shows about mesophiles.
Lab 2-9 The Effect of pH on Microbial Growth
Read the lab protocol found on pages 111-116 of Microbiology Laboratory Theory and Application by Leboffe and Pierce.
It’s important that you read the lab and data sheets to understand the scope of the lab activity.
Review ​The Effects of pH on Microbial Growth​ and use the information located there to answer the questions below.
1. On the pH scale below, indicate the 3 pH ranges (use brackets or double-ended arrow) that the 3 major pH
classifications of organisms can inhabit. Label each range (acidophile, neutrophile, alkaliphile) appropriately.
Chapter 14: Microbes in the Produce Aisle
Part One: The Case
In October, discussions in a small town in Louisiana centered on dramatic world events,
especially terrorism. However, an outbreak of pneumonia in 33 people from their community within a
single week changed the talk in the coffee shops and community center. Epidemiologists with the
state Department of Health began examining the patients for the most common causes of pneumonia
in adults: the bacterial species ​Streptococcus pneumoniae​, ​Mycoplasma pneumoniae​, and
Haemophilus influenzae​ and influenzaviruses and adenoviruses. The patients were negative for all of
these pathogens, and they contacted the Centers for Disease Control and Prevention (CDC). Basic
epidemiological strategy is to look for factors common to those infected but not common in nonvictims
that match the victims in age, geographical location, and similar factors. One factor the scientists
noticed was that patients were more likely to have shopped at a particular grocery store before
becoming hospitalized. This is puzzling, because while one might expect foodborne infections to be
related to a grocery store, what would be a link for pneumonia?
Imagine you are on the field team of the CDC sent to investigate this outbreak
1) What first steps would you take to solve this mystery?
2) How would you isolate and identify the cause of the pneumonia?
Part Two:The Mystery Pathogen
Epidemiologists call the first patient in an outbreak the index patient. Perhaps you first
assumed that an index patient transmitted the disease to other customers at the grocery store,
resulting in the outbreak. That would be a good start toward solving the mystery. However, this
pneumonia outbreak was atypical. As noted before, all of the most common pathogens of the lungs
(see Chapter 22) were absent in these patients. When epidemiologists began their investigation, they
found a surprise. An autopsy was performed on two patients who died. Lung biopsy samples were
positive for a Gram-negative bacterium, ​Legionella pneumophila​, as determined by a direct
fluorescent antibody test. This antibody test was used because culture of​ Legionella​ is slow (5 or
more days for the bacterium to grow) and technically challenging, requiring special media and
protocols. ​Legionella pneumophila​ is described starting on page 683 in the textbook.
3) Since ​Legionella​ is not spread from person-to-person, how could people in the grocery
store have been exposed?
4) What was the most likely route of entry (see pg 417) for the infected patients?
5) What is a likely mode of transmission (see Table 14.11, pg 430)?
Part Three:
Most of the patients in this outbreak had common risk factors for legionellosis, including
diabetes, chronic obstructive pulmonary disease (COPD), being a smoker, or being elderly. A key
discovery in the investigation occurred when ​Legionella​ was cultured from the water reservoir used in
the machine that misted produce in the vegetable aisle. Patients breathed the
bacterially-contaminated aerosol, resulting in exposure.
6) Why is Legionella considered an opportunistic infection?
7) How could contamination of the water reservoir be prevented in the future?
8) How are patients usually treated for legionellosis?
Resources:
https://www.cdc.gov/legionella/clinicians/diagnostic-testing.html
https://microbewiki.kenyon.edu/index.php/Legionella

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