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What Lives In Your Water? New MdBioLab Activity. G. Houston-Ludlam Graduate Student CBMG, UMCP. Our Activity. Step 1- Collect water samples Field trip or Homework Students should work in pairs Will require a “collection kit” Clean plastic bottles Gloves

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what lives in your water new mdbiolab activity

What Lives In Your Water?New MdBioLab Activity

G. Houston-Ludlam

Graduate Student

CBMG, UMCP

our activity
Our Activity
  • Step 1- Collect water samples
    • Field trip or Homework
  • Students should work in pairs
  • Will require a “collection kit”
    • Clean plastic bottles
    • Gloves
    • Ziplocs for ice and containment of sample

http://ian.umces.edu/imagelibrary/albums/userpics/10025/normal_iil_ian_bf_395.JPG

our activity1
Our Activity
  • Step 2- Filter water samples and culture overnight
    • 2 different volumes
      • 10 ml
      • 100 ml
      • Allows for best opportunity to get a countable plate of 20-60 colonies

http://www.umesc.usgs.gov/aquatic/drug_research/capabilities.html

our activity2
Our Activity
  • Step 3- (Next Day) Count Colonies

Example of bacterial growth on selective media. Photo courtesy of Hornor Lab, Anne Arundel Community College, Arnold, MD.

equipment setup
Equipment Setup
  • Completely assembled filtration apparatus
  • Water samples in ice bucket
  • Field data sheet
  • Sterile 10 ml syringe or pipet
  • Beaker with ethanol holding forceps
  • Sterile paper filter
place filter 1
Place Filter 1
  • Peel cover off filter (best done by instructor or partner)
  • Grab edge with sterilized forceps
place filter 2
Place Filter 2
  • Place paper filter grid side up on top of metal screen
  • Paper must completely cover screen to get proper filtration
reassemble filtration apparatus
Reassemble Filtration Apparatus
  • Place filter funnel on top of paper filter
  • Clamp glassware in place
10 ml sample
10 ml Sample
  • Wet filter with 10 ml sterile, distilled water
    • Water removes static from syringe
  • When the water has suctioned through filter, apply 10 ml of water sample to filter
wash filter funnel
Wash Filter Funnel
  • With clean syringe, wash the sides of the funnel to get any splashes
remove filter
Remove Filter
  • Unclamp filter funnel
  • Grab edge of filter with sterile forceps and break vacuum seal
place on plate
Place on Plate
  • Hold plate tilted downward and away
  • Place filter at bottom edge of plate
  • Roll onto media to minimize bubbles
  • Cover and incubate 24 hrs
repeat for 100 ml
Repeat for 100 ml
  • Place new filter on filtration apparatus
  • Wet filter and suction through
  • Pour 100 ml into funnel
  • Wash sides of funnel
  • Place filter on media
after incubation
After Incubation
  • This is what the students will see after a 24 hour incubation at 41˚C (chicken body temperature)
  • Left-hand plates came from Patuxent River
  • Right-hand plates came from Warehouse Creek off South River
  • Top plates are 10 ml, bottom plates are 100ml samples