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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. 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

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  1. What Lives In Your Water?New MdBioLab Activity G. Houston-Ludlam Graduate Student CBMG, UMCP

  2. 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

  3. 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

  4. 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.

  5. 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

  6. Place Filter 1 • Peel cover off filter (best done by instructor or partner) • Grab edge with sterilized forceps

  7. Place Filter 2 • Place paper filter grid side up on top of metal screen • Paper must completely cover screen to get proper filtration

  8. Reassemble Filtration Apparatus • Place filter funnel on top of paper filter • Clamp glassware in place

  9. 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

  10. Wash Filter Funnel • With clean syringe, wash the sides of the funnel to get any splashes

  11. Remove Filter • Unclamp filter funnel • Grab edge of filter with sterile forceps and break vacuum seal

  12. 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

  13. 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

  14. 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

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