Lab 18: Enterotube

Enterotube test is only for suspected Entero- (intestinal) bacteria

Lab 25: pBLU

lawn

white colonies

blue colonies

                                                lawn                           NG                    NG

                                                Lawn               White Colonies       Blue Colonies

Lab 24: Bacteriophage Titer

Lab 28: Blood Smear

1. Leukocytes= White blood cells (WBC’s)= cells of body’s immune defenses

Non-specific Leukocytes=WBCs of innate immunity= nonspecific cells that help combat foreign invaders 

Includes: neutrophils, eosinophils, basophils, monocytes, and natural killers (NK) cells

Specific Leukocytes= WBC of adaptive immunity= specific cells that destroy specific foreign invaders

Includes: T and B lymphocytes 

-in this lab we look at blood smears and identify different leukocytes 

1. Neutrophils= multilobed nucleus and granules

2. Lymphocytes – specific subtypes cannot be differentiated without special staining 

-smallest cells, large nucleus- makes cell look like it is crescent moon

3. Monocytes= largest cells, nucleus is U or kidney bean shape 

4. Eosinophils= red granules- 2 lobed nucleus 

5. Basophils- purple granule with obscure nucleus 

• WBC in body from most to least=  Never Let Monkeys Eat Bananas 

****KEEP IN MIND- These photos were not taken in class... my photos didn't turn out well.  These photos are one's I found on the web.  I recommend you check out the photos provided by K. Gard on WebCt... but for general idea, these pictures should help you out : )*****

1. Neutrophils

  

2. Lymphocytes 

3. Monocytes 

4. Eosinophils 

5. Basophils

(Lecture) "The Boy in the Bubble" Movie

1. What is SCID, and how does it lead to death?

Severe Combined Immuno Deficiency=SCID

-a genetic disease 

-no immune system- so any germ can kill you

-only affects males

2. What was one doctor’s bold new idea to address SCID in the 1970’s?

 Place infants at risk for SCID in a sterile plastic bubble immediately after birth until they can be tested for disease

 If they end up having disease then give them a bone marrow transplant for new immune system

3. Why did the plan not work for David?

-bc David’s older sister was not a bone marrow match 

4. When David was 18months old, what choices faced the doctors regarding his condition?

- they thought about removing David from bubble and letting disease run its natural course, because there were no treatments at the time and the bubble was originally a temporary not permanent structure 

-they decided to leave him in the bubble 

5. How did David become a research subject, and did this benefit science?

- bc doctors performed experiments/tests on him to understand the disease

-plus they kept him in a bubble

- Yes, research benefited science, learned:

- how SCID was caused

- that viruses can lead to cancer

- what germ free blood was like

- what isolation does to children 

6. How did the doctors control his behavior when he tired to poke holes in the bubble?

- They made him aware of the dangers presented by the outside world

-ie. they told him how microbes would kill him

7. Why did David’s special suit not work?

- bc he was afraid of getting in suit bc he didn’t want germs to get him

8. As David grew older and more aware of his situation, how did he respond psychologically?

- Depression, anxiety, distant, fearful, nightmares

9.What new hope was there for David at age 12?

- Experimental bone marrow transplant from non match ie. his sister could now donate her bone marrow even though it wasn’t a perfect match

10, How did David die?

-after receiving experimental bone marrow transplant, his body grew various cancerous tumors because his sisters bone marrow contained a dormant virus

-lead to discovery that viruses can cause cancer 

Lab 16: Catabolism of Proteins

 Tests For the Catabolism of Proteins

1. Hydrogen Sulfide (H2S) Test/Indole Test/ Motility (SIM)

a. Hydrogen Sulfide (H2S) Test

b. Indole Test

c. Motility Test 

2. Phenylalanine Deaminase Test

3. Rapid Ornithine Decarboxylase Test 

4. Urease Test 

1. Hydrogen Sulfide (H2S) Test/Indole Test/ Motility (SIM)

a. Hydrogen Sulfide (H2S) Test

-tests for enzyme cysteine desulfurase-bacteria with this enzyme can break down amino acid cysteine

- growth medium- SIM medium agar 

Positive= black

Negative= medium remains yellow or black line forms just below surface

Hydrogen Sulfide (H2S) Test

Positive=black

Neg= yellow or yellow with black line 

Hydrogen Sulfide (H2S) Test

b. Indole Test

-tests for enzyme tryptophanase- if bacteria has they can break down tryptophan 

- growth medium=SIM medium

- Positive= pink strip

-Negative= white strip (no change)

Indole Test

-note #22 (the middle strip) is the ONLY NEGATIVE =WHITE strip

Indole Test 

Positive=pink  

Indole test

 positive-= pink

 Indole Test

#22= negative= white

#27= positive= pink 

c.Motility Test 

+= black or yellow with fuzzy growth 

- = yellow with 2d ribbon growth

*note-when reading motility- ignore the little black line just below surface?

Motility Test= 

 Negative=2d Ribbon 

 Motility Test

Negative= 2D ribbon 

 Motility Test

Positive= Black 

Motility Test

Positive= Yellow and Fuzzy 

(think pipe cleaner)

 Motility Test

Positive= Yellow and Fuzzy

1. 
   

2. Phenylalanine Deaminase Test 

-tests for enzyme phenylalanine deaminase

-growth medium= phenylalanine agar

-reagent= ferric chloride (FeCl3)

-after incubating, the reagent ferric chloride is added to the medium

+ test= green = -phenylpyruvic acid reacts with ferric chloride and turns green

- test= no change- medium remains yellow

Phenylalanine Deaminase Test 

Phenylalanine Deaminase Test

Positive= green 

Phenylalanine Deaminase Test

Negative= yellow= no color change 

3. Rapid Ornithine Decarboxylase Test

-tests for enzyme ornithine decarboxylase

- removes carboxyl group from amino acid ornithine

-growth medium= rapid ornithine medium

Positive test= purple

Negative test= grey/clear 

Rapid Ornithine Decarboxylase Test

#32= negative= grey/clear

#16= positive=purple 

1. 
   

1.  

4. Urease Test

test for enzyme urease

growth medium= urea agar

Positive test= fushia= presence of urease causes the formation of basic products ammonia and ammonium hydroxide

Negative test= medium remains yellow= medium stays acidic

Lab 15: Catabolism of Carbohydrates -MR-VP AND Citrate Test

MR Test
Growth medium= MR-VP broth
Reagent= methyl red indicator
Positive= red
Negative= yellow
Uninoculated= yellow

Positive MR test =RED

 Negative MR test= Yellow

 Uninoculated Tubes= yellow 

VP Test
growth medium= MR-VP broth
reagent= alpha-naphthol and potassium hydroxide (KOH)
Positive= crimson
Negative= copper
Uninoculated= yellow (see above)

VP test 

 Positive VP= crimson 

 Negative VP= copper 

CItrate test 

-tests for enzyme citrate permease 

Growth medium= Simmon’s citrate agar -contains bromomythmol blue= pH indicator that is green= neutral and blue= basic

Positive Citrate test= blue- bacteria contains enzyme citrate permease 

Negative Citrate test= green- bacteria does NOT contain enzyme citrate permease 

Citrate Test 

Negative Citrate= Green 

Positive Citrate= Blue

note how the one on the right is mostly blue with some green, 

this is still positive bc it is turning blue