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- 1. tube has loop, wire goes through all chambers, inoculates all chambers.
- 2. color diagram for all positives
- 3. tests enteric bacteria on various sugar fermentations, sulfur reduction , indole production, citrate utilization, etc.
- 4. looks like long multicolored tube with that wire that goes through entirety of tube
- : 12 tests at once, touch tip on bacteria and pull through media. Results coded for genus and species
- 1. can do for anything
- 2. isolate colony, put in saline, pipette drops into wells
- 3. incubate for 24 hrs , find #, tells genus and species, 20 tests at once
- 20 tests at once, mix saline solution with bugs pipette into wells. Results coded for genus and species
BBL Crystal kits
- 1. put bug in saline, dump in trough
- 30 tests at once, mix saline solution with bugs inoculate- score sheet provides biocode number-software program
- specialized blood agar
- 90 tests at once, computer reader tells you +/- and tells you what the bug is. Expensive, specialized agar. Must be a certain amount of days old. Efficient and very accurate.
- can be used for anything
- relies on combination of antibodies, antigen
ELISA based tests
- 1. takes salmonella , will react if +
- 2. tests for enzyme linked salmonella
- 3. does many tests at a time
salmonella 1-2 tests
- 1. for smaller masses of testing at a time
- salmonella will start swimming around, white crescent forms for +
- 2. + = yellow, cloudy yellow
- 1. sterile premoistened swab, twists, pushes swab into luciferan, luciferase, ATP (from cell)
- 2. ATP will react w/ L's and glow bc L's are bioluminescences
- fruit roll up version of petridish
- 2. Pro: less space, more convenient
- 1. generally used to grow stock cultures that can be refrigerated after incubation and maintained for several weeks
- 2. many differential media are agar slants
- vials are designed for storing biological material, human or animal cells, at temperatures as low as -196°C but should be used only in the
- gas phase of liquid nitrogen.
- 1.Individual beads may be removed without thawing the whole sample
- 2. involved in cryogenics
- 1. is a dehydration process typically used to preserve a perishable material or make the material more convenient for transport.
- 2. Freeze-drying works by freezing the material and then reducing the surrounding pressure to allow the frozen water in the material to sublimate directly from the solid phase to the gas phase
- 1. the global bioresource center
- 2. American Type and Culture Collection
- 1. must be careful due to bioterrorism
- Ground Transport: do NOT use personal vehicle, could be personally viable if it is not completely safe
- Packaging: layers of protection
- 1. put it in tube, absorbent material, bubble wrap, put it in 2nd tube,
- 2. must have paper w/ material have federal government sign for it
- 3. mail/parcel services: when name on shipping label, responsible until someone else signs for it
- 1. CDC , NIH identifies these, also DOT does
- 2. must have permit if restricted
- 3. CYA (cover your ass with paperwork)
- 1. (bacteriological Analytical Manual)
- 2. standardizes all tests
- 3. usually assigns numbers to each test
1. What is source is about how to test food products?
Compendium of Methods for the Microbiological Examination of Foods
ASM Manual for Evaluation of Clinical Samples (multiple volumes)
medical version of BAM
Methods for General and Molecular Bacteriology
1. to catch up on basic micro stuff if have bio background
1. what represents the best current practice of American water analysts.
- 1. Standard Methods for the Examination of Water and Wastewater
- 2. APHA?
- 1. American Type and Culture Collection
- 2. tells what biosafety level
- 3. info on bugs- all bugs can be ordered
- the manual in class that had detailed information about specific species of bacterium. Real collection is massive, with many many volumes
- 5 volumes
- 1. Material safety data sheet
- (must have 1 for every single chemical)
Biosafety in Microbiological and Biomedical Laboratories
- 1. (CDC/NIH)
- 2. what are minimum requirements needed in lab based off of what materials you're working with
- 3. : what min requirement you have to have in your lab. Eye protection, gloves, emergency showers, spill kits etc. OSHA uses this checklist for inspection
- 1. Loop bacteria onto slide.
- 2.Place small drop of 3% KOH 1” away from bacteria.
- 3. Drag small amount into bacteria and mix well for 30 sec. Gently lift loop off surface and look for “elastic string”
- If present= gram negative
- If not present= thick gram positive wall protected cell from KOH breakdown
Specific Laboratory Standard Operating Procedures (SOP's)
has all cautions in it
- 1. Clean slide with Bon-Ami and wipe dry with kemwipe.
- 2. Loop a colony near clear edge of slide.
- 3. 1-2 drops Congo Red on clump and mix. 4. 1 drop crystal violet mix and let stand 1 min.
- 5.Smear across slide using another slide. 6. 6. Allow to air dry rinse to observe.
- Thin pink line surrounded by white cloud indicates positive
Effectiveness of Disinfectants
- 1. Add 0.1 ml bug to sterile saline. Make a 0.1 ml spread plate.
- 2. Add 0.1 ml of disinfectant- This is a mix tube.
- 3. Add 0.1 ml to TBA at 0 min, 5 min, and 10 min- after each TBA tube is spread plate immediately. Incubate at 37 C
clear halo around bacteria=positive
use white card w printed circles add bacteria to card. Stir the antiserium and bacteria for 1 min. Positive=gray liquid