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 for our population of red wormsTo determine ld 50 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, CaCO 3, water Litmus paper Pesticide % 1-(4-CHLOROPHENOXY)-3,3- DIMETHYL-1-(1h-1,2,4- TRIAZOL-1-YL)-2-BUTANONE)

Experiment layout 10 worms added to each jar Worms allowed to adjust to new soil habitat pesticide added after week 1 Data collected weekly worms sorted and analyzed for lesions, swelling, writhing, Number on 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

pH of each soil was taken after soil was mixed and again after the fungicide was added Initial pH of the soils were about 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