Final Project

NUMBER	PRODUCT	DISPLAY TEMPERATURE		STORE NAME OR DESCRIPTION  OF PRODUCT LOCATION	COMMENTS  (Determine & Discuss if Temperature meets  antimicrobial standards for low temperature preservation and reduction of microbial activities)
		(oF)	(oC)
1.	Gallon of Fresh Milk	35 °	2 °	PUBLIX  In the open refrigerator next to sour cream	According to online sources of education (NC State University), milk should be stored at 32- 45 °F. Bacteria will no grow at this parameters of temperature. Meets standards.
2.	Fresh Orange Juice	37 °	3 °	PUBLIX Open refrigerator next to Natural Juices	Florida Citrus indicates to all customers that orange juice should be stored at 40 ° F or below to  avoid the loss of its quality taste. Meets standards.
3.	Any 2 fruits (Name Fruits)	39 ° 39 °	4 ° 4 °	PUBLIX Red Apples  Strawberries	Iowa State University recommends to keep apples at a temperature near 32ºF and a relative humidity between 90 and 95 percent. This will ensure they are kept fresh for longer. For strawberries the Geroge Mateljan Foundation recommended to keep them at an optimum temp of 36 to 38º F.
4.	Yogurt (any Brand/Flavor)	39°	4°	WINN DIXIE La Yogurt Open refrigerator  near cream cheese	The recommended temperature to keep yogurts at is 35 to 40º F, this will inhibit the yogurt from spoiling.
5.	Eggs (Specify size)	18 °	-8 °	All sizes are stored side by side on the Dairy section. Publix store	According to the egg safety center, eggs should be  kept at a temperature of 45 °F or below to avoid bacterial growth. Thus indicating that it meets the standards.
6.	Chicken (Leg Quarters, Drumsticks, or Wings)	28 °	-2 °	Chicken is kept refrigerated together in separate packages  on the Meat area, poultry section.	Food safety Government site indicates that all chicken should be stored at a temperature lower than 40ºF to avoid the growth and reproduction of  bacteria that is already in the raw chicken.
7.	Deli (just one, by Name)	35 °	2 °	WINN DIXIE Virginia Ham	Ham is also stored at a temperature les than 40ºF because higher temperatures stimulate the rapid growth on meats and poultry.
8.	Whole Cantaloupe	38 °	3 °	WINN DIXIE Open refrigerator next to pineapples	The optimal storage temperature for ripe cantaloupe is 36 to 41º F, said the ANR University of California, to avoid the growth of bacteria such as E.coli and salmonella. Meets the standards.
9.	Whole Rotisserie Chicken	27 °	-3 °	PUBLIX Open refrigerator next to the ground beef	Rotisserie chicken is stored in an air sealed container for storage, since it was previously cooked at really hight temperatures. To avoid spoilage and bacterial growth the temperature stored at should be less than 40º F as recommended.
10.	Meat, Beef, or Steak	27 °	-3 °	WINN DIXIE Open refrigerator near the Ribs	Food services of America recommends the costumers to store the meat at  28F and 32F, since bacteria will grow rapidly at 40º F.
11.	‘Parmalat’ Brand Milk	77°	25°	Store environment temperature.  Publix store; Line 5 on the same shelf of coffee and canned milk, etc.	Parmalat is stored at room temperature (cool environment) up to 6 months, which is the shelf life when it is still unopened. Once opened it needs to be refrigerated at 35.6 to 37F and consumed within 7 days, to avoid the bacterial infection and food poisoning.
12.	Hospital Rooms	50-70 °	10-21 °	Hialeah Hospital	Hospital rooms are kept at a low temperature to avoid more growth of microorganisms, or keep microorganisms that are already present, low. Static.

Test 3 Review

 TOPICS FOR TEST #3

1. BACTERIAL SHAPES & ARRANGEMENTS:

Name and describe major
Coccus (single)
Coccobacillus - (look oval )
Diplococci - (Two together)
Streptococci -(chain)
Staphlylococci - ( Cluster or group)
Sarcinaecocci (8 together)
Tetradcocci (4 together)
Bacillus - Single
Diplobacilli
Streptobacilli
Spirillum - (Spiral Shape)
Vibrio - Curve Baccillus
Spirochete -  Like Snake (must be seen live)
 

a. Shapes used to identify Bacteria

 

b. Arrangements for Bacterial identification

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2. DIFFERENTIAL STAINING

a. Describe and contrast Differential Staining with Simple Staining

- Differential Staining emphasizes bacterial diversity with color
 - using 2 basic dyes with contrasting colors to stain bacteria differently.
- Differential staing is about Highlighting differences between bacteria 
 - Each Differential Staining procedure uses its unique pair of basic dyes. 
-  Differential staining results in positive / negative results .
- Most frequently used differential stains are 
          1.  Gram Stain - to distinguis different kinds of bacteria
          2.  Acid-Fast stain -  distinguishes Mycobacterium 

- simple Staining uses just one dye:( a basic dye ) it highlights the specimen with color in contrast to a clear background. 
- primary purpose is to highlight the entire microorganism so tht cellular shape and basic stucture are visible.

(Special Staining)  is used to isolate specific parts of microorganisums...  such as endospores and falgella and the presence of capsules..

 b. Learn reagents and Chemical Groups of Dyes used

      (pairs of Basic Dyes for diff. staining)

      Crystal violet (purple) ---- Safranin (red)      ------>(if primary Stains stays purple +)

      Carbol Fuchsin ----------------------- Methylene Blue  ------ used in acid fast staining 

      Malachite Green  -------------------Safranin ----- used in endospore (spore) staning 

c. Recall steps and what each step does to the cell

      

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3. THE GRAM STAIN

a. List the steps and describe final results.

b. Identify Mordant, Decolorizer, Primary, and Counter stains 
                 1. Primary die  --- (colors everything)
                              1b. Apply MORDANT ----- it fixes the primary stain , not a dye,

                                                            no color change 
2.  Deodorizers ----either Alcohol, Acitone, Acid      

                                   1. alcohol--> ethanol or proponal

                                   2. Acetone ---(acetone + alcohol 1:1) 
                                                         dehydrates cell,
                                                         never pure ...
                                   3. Acid ------- (Acid 3% + Alcohol 97%),
                                                                     deodorizes &   "destroys", 
                                                                     usually not used too strong

                                   4. Water------ rarely used 
 3. Apply Counter Stain---- ( Safranin contrast for crystal violet )

c. Determine how specimens look after each step of Gram Stain
       

d. Review final results of Gram Stain

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4. WHY GRAM STAIN?

Learn the Explanations for Gram Reaction:

Peptidoglycan: Occurrence, Location, and Thickness

Peptidoglycan – consists of repeating disaccharide attached to Polypeptides to form lattice

Peptidoglycan found in cell wall of “Eubacteria”

Prokaryotic cells - Archaea does not have peptidoglycan in cell walls.

 Gram positive = wall has 95% peptidoglycan -

 Gran Negative = wall has 15% peptidoglycan + 80% outer membrane

      Outer membrane – full of fat (Lipids) – Liproteins,

                                                                               phospholipids,

                                                                                lipopolysaccharide,

                                                                                periplasmic space

          Periplasmic Space –(gap) which mean less material to stain – contains high levels of degradative enzymes and transports proteins..

 

Crystal Violet-Iodine Complex –

                     is the primary dye for gram stain

                     It dyes both gram + and Gram –

                     Its retained by Gram + because wall has lots 95% of peptidoglycan

                     It is washed out from gram- because has only 15% peptidoglycan.

 

Result

Gram + purple (visible)

Gram – (invisible – until you do counterstaining with safranin- turns pink red)

 

Components and Chemistry of Bacterial Cell Wall

  - Teichoic Acids – only in gram + , attached to peptidoglycan only

  - Lipoteichoic acids - only in gram + , attached to peptidoglycan and  plasma membrane

 -  Basic Dyes

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5. GRAM PROFILE AND COMPARISON OF GRAM POSITIVE

AND NEGATIVE BACTERIA IN TABLE 4.1, PAGE 87

a)  Basal Bodies                      e) Phospholipids

b)  Outer membrane               f) Lipo-Polysaccharides

c)  Lipid Content                     g) Types of Lipids

d)  Flagella Structure              h) Periplasm

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6.     EXOTOXINS

a) What are they and where do they come from?

b)  Which microbes make them?

c) How Toxigenic are they and how do they work?

d) When can they be used for Immunological benefits?

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7.     FUNCTIONAL GROUPS OF EXOTOXINS

a) Identify the 3 Functional names of Exotoxins

b) Describe their properties and targets

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8.     STRUCTURAL TYPES OF EXOTOXINS

a)  Name the 3 Structural Types of Exotoxins

b)  Describe their properties and how they work

c)  Explore specific examples and list Bacteria associated with each

d) Study the bacteria, Toxin, and Symptoms of Toxic Shock Syndrome

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9.    BACTERIAL ENDOTOXINS

a)  Recall major properties of Bacterial Endotoxins

b)  Compare their characteristics to Exotoxins

c) Describe Symptoms of Endotoxins and explain how they are provoked

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10.    BACTERIAL TOXIGENICITY

a)  Define Toxigenicity?

b)  Define bacterial Virulence and identify some virulence factors

c) Compare Properties and Toxigenicity for:

      Exotoxin and Endotoxin

      Bacteremia and Toxemia

      Streptolysin S and Streptolysin O

      Leukocidin and Hemolysin

      Virulence and Non-Virulence

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11.     TOXOIDS

a)  What are Toxoids?

b)  Where do they come from and how are they formed?

c)  Explain their clinical roles as vaccines

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12.     ANTI-TOXINS

a)  What are Anti-toxins?

b)  Where do they come from and how are they formed?

c)  Explain their role in Immunity

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