What Lives In Your Water? New MdBioLab Activity
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What Lives In Your Water?New MdBioLab Activity
G. Houston-LudlamGraduate Student
CBMG, UMCP
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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
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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
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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.
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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
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Place Filter 1
• Peel cover off filter (best done by instructor or partner)
• Grab edge with sterilized forceps
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Place Filter 2• Place paper filter grid
side up on top of metal screen
• Paper must completely cover screen to get proper filtration
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Reassemble Filtration Apparatus
• Place filter funnel on top of paper filter
• Clamp glassware in place
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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
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Wash Filter Funnel
• With clean syringe, wash the sides of the funnel to get any splashes
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Remove Filter
• Unclamp filter funnel• Grab edge of filter with
sterile forceps and break vacuum seal
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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
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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
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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