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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PROJECT 6

WORKSHEET OF ANTI-MICROBIAL AGENTS FOR 5-MEMBER GROUP

Group Number: ________2___________

#	NAME OF ANTIMICROBIAL AGENT/PRODUCT	CATEGORY OR TYPE OF ANTIMICROBIAL AGENT	ACTIVE INGREDIENT IN ANTIMICROBIAL AGENT		MODE OR MECHANISM OF ANTIMICROBIAL ACTION
			INGREDIENT’S NAME	AMOUNT
1.	Purell	Sanitizer Antibacterial	Ethyl alcohol (Ethanol)	62%	Alcohol denatures microbial proteins
2.	Biaxin	Antibiotic	Clarithromycin a macrolide antibiotic	250 mg per 5mL oral suspension	Inhibits bacterial protein synthesis
3.	Contact Lens Disinfectant	Disinfectant	Hydrogen peroxide	3%	Hydroxyl free radicals produced by hydrogen peroxide destroy membrane lipids of microbe
4.	Listerine	Antiseptic	Eucalyptol Menthol Tymol	0.092%	Bacterial cell wall destruction, Bacterial enzymatic functions inhibition
5.	Zithromax	Antibiotic	Azithromycin	250mg	Azithromycin binds to rRNA of a bacterial ribosomal subunit, it blocks protein synthesis by inhibiting peptide translocation
6.	Anti-Bacterial Soap	Antibacterial	Triclosan	15%	Triclosan inhibits synthesis of fatty acids. Triclosan damages the cell walls of bacteria, slowing their growth so that the bacteria eventually die.
7.	Tilex	Antibacterial disinfectant	Sodium hypochlorite	2.40%	It causes proteins to lose their structures it is considered an oxidizing agent that prevents the cellular enzyme system from functioning.
8.	Lysol	Disinfectant spray	Alkyl (50% C14, 40% C12, 10% C16) dimethyl benzyl ammonium saccharinate, Ethanol	0.10% 58.00%	The ethanol in Lysol denatures protein.
9.	Colgate Toothpaste	Anti-cavity	Sodium monofluorophosphate	0.76% (0.15% w/v fluoride ion)	When bacteria in the mouth combine with sugars they produce acid. This acid can erode tooth enamel and damage our teeth. Fluoride can protect teeth from demineralization that is caused by the acid.
10.	Amoxil	Antibiotic	Penicillin	500mg	inhibits cell wall sythesis by binging antibacterial protein
11.	Cipro	Antibiotic	Ciproflaxicin hydrochloride	500mg	Affects the ability to grow by interfering with bacterial enzymes which are nessacery for bacteria to copy and repair DNA
12.	Levaquin	Antibiotic	Levoflaxin	700mg	Affects the ability to grow by interfering with bacterial enzymes which are nessacery for bacteria to copy and repair DNA
13.	Scope	Antibacterial, Antiseptic	Cetylpyridinium Chloride	0.05 - 0.10%	Contact with bacteria occurs by the disturbance of the cell membrane, inhibition of cell function, seepage of cell contents, and eventually cell death
14.	Speed Stick Gel Deodorant	Antiperspirant Antiseptic	Aluminum zirconium tetrachlorohydrex	10%, 15.4 and 18 %	Functions by diffusing into the sweat gland and forming a colloidal "plug" which limits the flow of sweat to the skin surface. The plug is gradually broken down and normal sweating resumes.
15.	Clorox	Disinfectant	Sodium Hypochlorite	6%	Hypochlorite, the active ingredient of clorox attacks essential bacterial proteins by causing proteins to lose their structure and form large aggregates. This inhibits the bacterial growth and ultimately kills the bacteria.
16.	Hydrogen Peroxide	Disinfectant Antiseptic	Hydrogen Peroxide	3%	Oxidation of cellular components (membrane lipids), growth inhibition
17.	Fabuloso	Cleaning and foaming agents	Sodium Dodecyl Benzene (linear) C9-11 Pareth-8 Sodium Laureth Sulfate	1%	Not antimicrobial, cleaning agents
18.	Gillette Antiperspirant Gel	Antiseptic	Aluminum Zirconium Octachlorohydrex Gly	16%	Aluminum salts are antimicrobial agents, they inhibit microbial growth by denaturing bacterial protein
19.	Dishwashing Liquid	Antibacterial	Triclosin	10%	Triclosan inhibits synthesis of fatty acids. Triclosan damages the cell walls of bacteria, slowing their growth so that the bacteria eventually die.
20.	Alcohol Swab	Antiseptic	Isopropyl	70%	Alcohols dehydrate cells, disrupt membranes and cause coagulation of protein
21.	Aquafresh	Anti-cavity	Sodium monofluorophosphate	0.15% w/v fluoride ion)	Both compounds produce ionic fluoride, which blocks glycolysis and exchanges with the hydroxyl group of the hydroxyapatite crystal of enamel. As a result, oral bacteria produce lesser amounts of acidic compounds to become more insoluble.
22.	Tide Detergent	Detergent	Borax (sodium borate)	20g/liter	Borate is a bacteriostatic and fungistatic. It inhibits bacterial and fungal growth and proliferation.
23.	Betadine	Antiseptic	Povidone iodine	10%	Impairs protein synthesis and alters cell membranes.
24.	Formaldehyde	Drying agent	Formaldehyde	10%	Inactivate proteins by forming covalent cross links with several organic functional groups on proteins
25.	Doxycycline	Antibiotic	Doxyeyclyline	200mg	lopophilic pass through lipid bilayerof bacteria blocking the binding of RNA and DNA inhibiting protein synthesis
26.	Savlon	Antiseptic Biocide	Cetrimide Chlorhexidine	3.0% 0.3%	Cetrimide-Destroys and damages cell membrane by lowering tension of surface. It causes microbial protein denaturation. Chlorhexidine-Binds to cell wall and destabilizes by osmosis causing it to rupturewhich causes fluid to leak leading to lysis and death.
27.	Mitchum Antiperspirant		Aluminum Sesquichlorohydrate (anhydrous)	25%	Antiperspirant
28.	Vinegar	Antibacterial	Acetic Acid	5%	The acid kills bacteria and viruses by denaturing (chemically changing) the proteins and fats that make-up these.
29.	Dynacin	Antibiotic	Minocycline Hydrochloride	50, 100 and 200 mg	Inhibits protein synthesis and thus bacterial growth by binding to 30S and possibly 50S ribosomal subunits of susceptible bacteria
30.	Neosporin	Antibiotic	Bacitracin zinc, neomycin sulfate, and polymyxin B sulfate	BZ: 400 units, NS: 3.5mg, PBS: 5000 units	Actively transported across the bacterial cell membrane, binds to a specific receptor protein on the 30 S subunit of bacterial ribosomes, and interferes with an initiation complex between mRNA (messenger RNA) and the 30 S subunit, inhibiting protein synthesis. DNA may be misread, thus producing nonfunctional proteins; polyribosomes are split apart and are unable to synthesize protein

References:

http://dailymed.nlm.nih.gov/dailymed/lookup.cfm?setid=b5bae1e7-2ee2-48e6-a50a-194527a19d39

http://www.nlm.nih.gov/medlineplus/druginfo/meds/a692005.html

http://aac.asm.org/content/45/7/2038.full

http://www.ncbi.nlm.nih.gov/m/pubmed/2133450/

http://www.pharmaceutical-drug-manufacturers.com/pharmaceutical-drugs/antiseptics.html

http://www.infectioncontroltoday.com/articles/2000/11/a-comparison-of-commonly-used-surface-disinfectan.aspx

http://www.patient.co.uk/health/antibiotics-leaflet

http://www.life.umd.edu/classroom/bsci424/Chemotherapy/AntibioticMechanisms.htm

http://www.tufts.edu/med/apua/about_issue/agents.shtml
http://ingredientpal.com/ingredients-of-listerine-mouthwash-in-cool-mint

http://www.medicines.org.uk/emc/medicine/1477/SPC/Zithromax+Capsules,%20+Suspension
http://www.drugs.com/search.php?searchterm=aquafreshAnnex

Mechanisms of antimicrobial action

TOPICS FOR TEST #3 1. BACTERIAL SHAPES & ARRANGEMENTS: Name and describe major a. Shapes used to identify Bacteria b. Arrangements for Bacterial identification 2. DIFFERENTIAL STAINING a. Describe and contrast Differential Staining with Simple Staining b. Learn reagents and Chemical Groups of Dyes used c. Recall steps and what each step does to the cell 3. THE GRAM STAIN a. List the steps and describe final results b. Identify Mordant, Decolorizer, Primary, and Counter stains c. Determine how specimens look after each step of Gram Stain d. Review final results of Gram Stain 4. WHY GRAM STAIN? Learn the Explanations for Gram Reaction:       Peptidoglycan: Occurrence, Location, and Thickness       Crystal Violet-Iodine Complex       Components and Chemistry of Bacterial Cell Wall       Teichoic Acids, Lipoteichoic acids, and Basic Dyes 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 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? 7.     FUNCTIONAL GROUPS OF EXOTOXINS a) Identify the 3 Functional names of Exotoxins b) Describe their properties and targets 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 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 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 11.     TOXOIDS a)  What are Toxoids? b)  Where do they come from and how are they formed? c)  Explain their clinical roles as vaccines 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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 TOPICS FOR TEST #3

1. BACTERIAL SHAPES & ARRANGEMENTS:

Name and describe major

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

b. Learn reagents and Chemical Groups of Dyes used

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

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

      Crystal Violet-Iodine Complex

      Components and Chemistry of Bacterial Cell Wall

      Teichoic Acids, Lipoteichoic acids, and 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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