Today's Lab Interpritation

Tuesday in lab we inoculate 3 tubes of Thioglycollate broth to determine the oxygen requirements of our unknowns. We inocullate the tubes with E. coli, Clostridium p. and our unknown. We also did a similare exercise with nutrient broth plates using E.C. and C.P. as our controls to compare our unknowns to.

Results:
E. coli: facultative
C.p: strict anaerobic
#30: facultative

Plate Interpritation:
E. coli: facultative
C.p: anaerobe
#30: facultative, with smooth edges and made the medium a slightly darker color.

From the inoculated broth or our working slants we where to make slides for staining and viewing to determine the shape of our unkown as well as the E. coli and Clostridium p.

My slides made from the slant were a little hard to examine. I remade 3 more from the broth but didn't have time to look at them. I did accidently stain all 3 rather than just one. Hopefully I can use these next week.

Microbe Shapes:
E. coli: bacillus
C.p: cocci clusters
#30: Dr. Belcher thought perhaps mine where cocci that formed chains. I couldn't tell becuase my smear was a little thick.

Next week it's gram staining action like you've never seen!

This Weeks Media

Eosin-Methylene Blue is used to select G- lactose fermenters. The selective agent is eosin and methylene blue. The differential agent is methylene blue and lactose. If the organism ferments lactose it is most likely G- and lactose +. Medium is used to grow enteric microbes.
E coli: Microbe tolerates EMB. Microbe ferments lactose. Microbe is most likely G-
S. Aureus: Microbe does not tolerate EMB. Microbe does not ferment lactose. Microbe most likely G+
Ps. aeroginosa:Microbe tolerates EMB. Microbe does not ferment lactose. Microbe most likely G-
Unknown #30: Microbe does not tolerate Eosin & MB. Does not ferment lactose. Most likely G+

Phenyl Ethyl Alcohol is a medium used to select G+ cocci. The selective agent is phenyl ethyl alcohol and there is not a differential agent. Organisms that show good growth on this medium are most likely G+.

E. coli: Microbe does not tolerate Phenyl Ethyl Alcohol. Most likely G-

Ps. aeruginosa: Microbe does not tolerate PEA. Most like G-

S. Aureus: Microbe tolerates PEA. Most likely G+

Unknown #30: Microbe tolerates PEA. Most likely G+

MacConkey Agar is a medium used to select G- lactose fermenters. The selective agent is bile salts and the differential agent is neutral red and lactose. Lactose + microbes appear pink-red. Lactose - microbes appear cream to colorless. This medium is used to grow G- enteric microbes. E. coli: Microbe tolerates bile salts & Crystal Violet dye. Does ferment lactose. Most likely G-

Ps. Aerug: Microbe tolerates bile salts & CV. Does not ferment lactose. Most likely G-

S. Aureus: Microbe does not tolerate Bile salts or CV. Does not ferment lactose. Most likely G+
Unknown #30: Microbe does not tolerate bile salts or CV. Does not ferment lactose. Most likely G+

Mannitol Salt Agar is a medium used to select mannitol fermenting staph microbes. The selective agent is sodium chloride and the differential agent is phenol red. Positive mannitol fermenters turn yellow.
E coli: Microbe does not tolerate Sodium Chloride. Does not ferment Mannitol. Most likely G-

Ps. Aerug: Microbe does not tolerate NaCl. Does not ferment Mannitol. Microbe most likely G-

S. aureus: Microbe tolerates NaCl. Microbe ferments Mannitol. Microbe most likely G+

Unknown #30: Microbe tolerates NaCl. Microbe ferments Mannitol. Microbe most likely G+

Summary:

Unknown #30 had similare growth and fermentation patterns as S. aureus. However, unkown #30 had slightly better growth and fermentation when compared to S. aureus.

Unknown #3o tolerates MSA & PEA and does not tolerate EMB & MAC. It tolerates Phenyl Ethyl Alcohol, NaCl and ferments Mannitol. This microbe is most likely Gram + and tolerates a salty environment.

Inerpreting Growth on Media

1. Purpose/ characteristics of media?
a. selective, differential, both
b. what are the agents
c. types of microbes typically isolated

2. Is there good growth?
Yes: "Microbe tolerates_(selective agent)_" No: "Microbe does not tolerate_(selective agent)_"

3. Is there a change in color?
Yes: "Microbe ferments/uses/poseses_(differential agent)_" No:"Microbe does not ferment/use/poses_(differential agent)_"

4. Microne is most likely "__G-/G+__"

Lab #6

Previously in lab we made a 4 section streak plate on 4 different types of growth medium.
We divided each plate into 4 section like a pie and in each section drew a line from 4 different microbe broths.
Media:
Eosin Methylene Blue Agar
Phenylethyl Alcohol Agar
MacConkey Agar
Mannitol Salt Agar

Microbes:
Unknown #30
E. coli
Staphyloccocus aureus
Psudomonas aeruginosa

E. Coli

Microbe:
Escherichia coli

Common Name:
E. coli

Oxygen requirements:
Facultative (Anaerobic or Aerobic)

Origin:
Intestines of mammals

Day 3 #30

This is my first Quadrant Streak plate and I'm rather proud of it. I had to redo it 3 times because I kept streaking from 1 instead of 3. My unknown #30 made neat little colonies in the middle.

Here are it's stats:

Growth Medium:
Nutritive Agar
Margin:
circular or punctiform
Consistency:
moist maybe slime
Pigment:
none
Odor:
smelled like dog poop (no joke)

Day 2 with #30

This is my working slant of the unknown #30. Next to it is my unknown in a differential broth. You can't tell from this picture, but my unknown is facultative. This means it utilizes both aerobic and anaerobic respiration.

Day 1 with Unknown #30

We made working slants of our unknown. That was pretty musch it. Hopefully I did it right and next week I'll see my microbe growing on the agar slant.

Flame the Loop

It is rather evident that this is my first Microbilogy class. Infact, it's my first Biology class since Highschool, which was over 10 years ago. In highschool Biology we touched apon amebas and looked at yeast eating sugar. We never did petri dishes and innoculations. Needless to say this is all Greek to me. Most of the genis names are Greek come to think of it. For example Staphalococcus means bunch of grapes in Greek.
The picture on the left was my artistict interpritation of the Sterile Innoculation process as outlined in my lab manual.
For those of you already in Micro or Lab Techs looking for a laugh, you will see that my ideas are a bit laughable.
I was thinking old school as to what the "loop" should look like.