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bIOMARKERS

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					 Biomarkers
by heather seavolt and
 stephanie mCPHERSON
    Types of
  biomarkers
• DNA, RNA or protein
  adducts and enzymes
• cytogenic markers
• tissue markers and whole
  body alterations
• behavioral markers
• measure toxicants in
  body fluids & tissues
• death of marker animals
     Ld50 & LC50
• Ld50 - median lethal
  dose (amount of a
  chemical in one dose
  that will kill 50% of
  the population)
• Lc50 - concentration of
  material in air that
  will kill 50% of the
  test subjects
Initial Purpose
• To observe any effects
  of the fungicide on
  the red worms

• To determine ld50 for
  our population of red
  worms
         SOIL
    ECOTOXICOLOGY
      EXPERIMENT
• Materials:
• 24 pint mason jars with lids
• Worms
• Soil mixture - composed of
  silica sand, kaolin clay, peat
  moss, CaCO3, water
• Litmus paper
• Pesticide - 0.88%
     1-(4-CHLOROPHENOXY)-3,3-
     DIMETHYL-1-(1h-1,2,4-
     TRIAZOL-1-YL)-2-BUTANONE)
     Experiment
       layout
• 10 worms        • Data
  added to each     collected
  jar               weekly

• Worms           • worms
  allowed to        sorted and
  adjust to new     analyzed for
  soil habitat      lesions,
                    swelling,
• pesticide         writhing,
  added after       Number on
    week 1          and below
                    surface
    fungicide
  concentration
      series
• Control series (3 jars)
• 6 series run with
  fungicide (3
  jars/series)
• first fungicide series
     4 ml fungicide :128 ml
        of water
• diluted 5 times for
  each successive 5 series
• 1st dilution: 1/2 the
  fungicide
  concentration of
  series 1 (2:128 ml)
• 2nd dilution: 1/4 the
  concentration of
  series 1 (recommended
  concentration)
• 3rd dilution: 1/8 the
  concentration of
  series 1
• 4th dilution: 1/16 the
  concentration of
  series 1

• 5th dilution: 1/32 the
  concentration of
  series 1
• Samples of lindsey and
  jason’s soil
• 1. sample from the
  copper pipe area
• 2. sample from site 1
• 3. sample from around
  suspicious oil
  container
     mortality
• Mortality is defined in
  this experiment as
  lack of movement in
  response to stimulus
  or absence, since the
  worms disintegrate
  rapidly
      Further
     objectives
• Additional objectives
  of this experiment:
• 1. examine effects of
  pollution on non-
  target organisms
• 2. Accuracy of results,
  if any, in real world
  situations
• 3. Consider effects of
  toxin on different
  organisms
• 4. How this experiment
  would be improved if
  carried out for a
  longer time and
  replicated
• 5. Ethical issues
  involved in animal
  experimentation
                                 data
                  Week
Number of worms alive 2              Week 3            Week 4            Week 5
                  Jar 1 Jar 2 Jar 3 Jar 1 Jar 2 Jar 3 Jar 1 Jar 2 Jar 3 Jar 1 Jar 2 Jar 3
Control Series         9 10 10           9 10 10           6     7     9     6     6 10
Original Series        6     4     8     6     5     8     6     5     5     5     5      5
1st Dilution           9 10 10           8 10 11           8 10 13           8     9 12
2nd Dilution          10 11        9     7     7     7     8     7     9     7     6 10
3rd Dilution           9     8 10        8     7 10        8     8     9     7     9      7
4th Dilution           8     0     9     8     0     9     9     0 10        8     0      9
5th Dilution          11     9     0 11        8     0     9     4     0     8     6      0
                       9
Soil - Copper Pipe area N/A N/A          7 N/A N/A         6 N/A N/A         5 N/A N/A
Soil - Oil Container 3 N/A N/A           3 N/A N/A         3 N/A N/A         2 N/A N/A
Soil- Site 1          10 N/A N/A         6 N/A N/A         6 N/A N/A         6 N/A N/A
Concentration vs. Percent Mortality
• pH of each soil was
  taken after soil was
  mixed and again after
  the fungicide was
  added
• Initial pH of the soils
  were about 6.0 - 6.5
• pH after addition of
  fungicide ranged from
  4.5 (in the more highly-
  concentrated soils) to
  5.5 (in the less highly-
  concentrated soils)
      results
• Slight trend in
  mortality rates and
  increased fungicide
  concentration

• in soil samples from
  jason and lindsey,
  mortality rate low
  except in soil taken
  from oil container
  site
   conclusions
• The highest fungicide
  concentration used (4
  times the suggested
  concentration)
  appears to have a
  fairly strong impact
  on the worms
• The soil sample from
  the oil container was
  not a good habitat for
  the worms since there
  was a fairly immediate
  drop in the worm
  count two weeks after
  the worms were
  introduced into this
  soil
• The other soils with
  varying
  concentrations of
  fungicides provided
  no worse a habitat for
  the worms than did
  the control series

• overall reproductive
  ability did not appear
  to be altered

				
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