Micro 5 Unknown Bacteria Identification

Spring 2019 Setup

* 9 flasks of BHI broth
	+ 50mL
	+ Use Media bottles??

* 12 flasks of Nutrient Broth (for instructor use)
	+ 50mL

Gram Negatives: Check if they are the same each semester\*\*\*\*\*

* *E. coli*
* *Citrobacter freundii*
* *Klebsiella pneumoniae*
* *Enterobacter aerogenes*
* *Salmonella enteriditis* (or whatever species in this genus that we have)
* *Shigella flexneri*
* *Proteus mirabilis*
* *Proteus vulgaris*
* *Providencia stuartii*
* *Morganella morganii*
* *Pseudomonas aeruginosa*
* *Alcaligens fecalis*

Gram Positives:

* *S. aureus*
* *S. epidermidis*
* *E fecalis*
* *Micrococcus luteus*
* *Corynebacterium xerosis* (this one was contaminated with Staph during the gram stain lab)
* *B. subtilis*
* *B. cereus*

### Lab 2: Streak Plating

* Incubated culture flasks
* Sterile culture tubes = 1/student
* sterile transfer pipettes

1 per student (plus 6 extra)

* NA plates
* MacConkey Agar (media info)
* PhenylEthanol Agar plates (Media Info)

### Lab 3: Isolation I

2 per student (plus 10 extra)

* NA plates

* Gram Staining
* Sterile water

### Lab 4 Isolation II

* Incubated plates
* Gram Staining
* Sterile Water

### Lab 5: Biochemical Testing I

(1 per student plus 5 extra):

* Incubated Plates
* Leftover media
* Sterile Water
* Triple Sugar Iron Agar slant (Media Info)
	+ 20 x 150 test tubes with 12mL TSI / slant. Make on slant rack. Note: Bigger Butts Please!
	+ Provide students inoculating needles
* MR-VP Broth (recipe here) (Media Info)
	+ 1 x 6 mL MR-VP culture tube student
* Simmon's Citrate Agar slant (media Info)
* Mannitol Salt Agar plate (media Info)
* Endospore Staining (see materials on Endospore lab) (Information)

### Lab 6: Biochem II

(1 per student plus 5 extra)

* Incubated Plates
* Leftover media
* Sterile Water
* Urea broth (recipe here) (media Information)
	+ Prepare Day-of:
	+ Pre-sterilize water and tubes but this broth cannot be heated
	+ make with sterile technique
	+ Prep sterile objects the day before and prep broth just before class
	+ 2mL broth tubes - more broth will make a slower reaction time
* SIM agar deep
	+ SIM Medium, 16 x 100 mm Tube, 8 ml Deep
	+ Read Here
* Nitrate Reduction broth (Information)
	+ Use premade medium
	+ **With durham tubes**
* Nutrient Gelatin (Media Info)
	+ 6mL in 16 x 100 tt
* Nutrient Agar Plates
* Glucose Phenol Red Broth (recipe here)
	+ Phenol Red HiVeg Broth Base = 16g/L
	+ Glucose (Dextrose) = 5g/mL
* Mannitol Phenol Red Broth (recipe here) (Media Info)
	+ Phenol Red HiVeg Broth Base = 16g/L
	+ Mannitol = 5g/mL
* Methyl Red and Barritt’s Reagents Sterile (Information)
	+ Transfer Pipettes = 1/ student
	+ Sterile Culture tube (methyl red test) = 1/student
	+ 3 Methyl Red Dropper Bottles (1 mL Broth w/3 drops)
	+ 3 x Barritt's Reagent A (4 drops)
	+ 3 x Barritt's Reagent B (2 drops)
		- See recipes in reagent library

### Lab 7 Biochem III

* Incubated Plates
* Leftover media
* Sterile Water
* Oxidase Reagent (see recipe)
	+ No filter paper. They add straight to plate
	+ Make for a 1x use and properly dispose of the unused amount
* Catalase
* Nitrate Testing reagents (A, B, and zinc)
	+ In Class Methods Here
* For SIM
	+ 3 x Kovac's Reagent
* For nutrient gelatin results
	+ Ice to cold incubate the tubes for 15 minutes

### Lab 8: Biochem IV

            Nothing further – but we may use some of the extras from above