fertilizer effects on acid rain stressed algae andrew haky 9 th grade, pittsburgh central catholic...
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Fertilizer Effects on Acid Rain Stressed AlgaeAndrew Haky
9th Grade, Pittsburgh Central Catholic HS1st Year in P.J.A.S.
RunoffPart of the water cycle and describes
the water that flows over a land surface.
Surface runoff occurs on land, typically creating a ‘watershed.’◦Materials that are transported on surface
runoff are fertilizers, petroleum, pesticides, herbicides, and salt.
◦Pollution can come from this runoff
EutrophicationCaused by an overabundance of nutrients
in an ecosystemNo limiting factor on algae populationsUncontrollable growth takes up resources
necessary for other organisms-oxygen.Limits biodiversityCan occur naturallyOccurs today by fertilizer run off
Serious Pollution Problem
Acid Precipitation
Worldwide problem Caused by pollution of
sulfur dioxide and nitrogen oxide
Often originates from smokestacks, vehicle exhaust, and burning fossil fuels
Average pH of 5.6
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An increase of acid rain in many aquatic systems reduces biodiversity.
Acid Rain in Pennsylvaniao Average pH of
precipitation in PA: 4.5
o Major problem due to industry
o The many abandoned coal mines in PA create a large problem for the water bodies in the area
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Nitrogen- found in ammonia used in fertilizers, causes algae populations to grow rapidly(eutrophication)
Phosphates are vital for cellular processes such as the synthesis of ATP and ADP. Both phosphates and nitrogen are normally limiting factors of algal populations.
The main ingredients in Miracle-Gro are Nitrogen, Phosphates and Potash.
Potash and
H2SO4
PotashPotassium oxide in
fertilizers93% used to make
fertilizersImportant for food
crops, gives them favorable qualities
Canada is the world’s leading producer
Sulfuric Acid Top product in
chemical industry Can be combined with
ammonia to create agricultural products
Corrosive, can bind with water droplets to create acid rain
ChlamydomonasUsed as a bio-indicatorGenus of green algaeCan create starchHas an Eyespot to orient itself
to lightTwo flagella, swims with a
breaststroke-like motion 10 μm in diameterLarge,crescent-shaped
chloroplastsFound in freshwater, soil,
oceans, and snow
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EuglenaProtistEuglenophyte because of chloroplasts20-300 μm in lengthEndocytosisFlagellum propels with whip-like motionCan also make an inchworm-like movement Has an Eyespot to orient itself towards lightFound in nutrient-rich freshwaterEffected by acidityAsexual, prokaryotic fission
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Purpose
Determine if fertilizers (Miracle-Gro) and acid rain (H2SO4 ) have an effect on algae populations (Euglena and Chlamydomonas) individually or in synergy.
Hypotheses
NULL- neither Miracle-Gro nor acidity will significantly affect algal population growth and they will not act synergistically to affect growth
PROCEDURE
1. 60 tubes were set up to receive equal light.2. Chlamydomonas was added to half the tubes
and Euglena to the other half. 3. Miracle-Gro and/or sulfuric acid was added to
the culture tubes in the following ratios:
4. using a micropipet in 0,0.5, & 0.05 mL volumes to 10 tubes each to both Chlamydomonas and Euglena samples.
5. 0.01x Sulfuric Acid was added using a micropipet to 5 out of every 10 tubes in 0.02 mL, but not the other 5.
6. Spring water was then added using a micropipet to make each tube 5 mL full.
Steps 2-5 created 30 tubes of Chlamydomonas. 15 tubes had a pH of 7 and the other 15 tubes had a pH of 5. For both of these groups, 5 tubes had a fertilizer concentration of 0x, 5 had a concentration of 0.001x, and five had a concentration of 0.1x. The same was also done using Euglena.
6.Each tube was marked with a black line so that it would enter the spectrophotometer the same direction every time.7.Wax paper was used to cover each tube. The tubes were each inverted 3 times.8. The spectrophotometer was allowed to warm up for 30 minutes at 430 nm.9.Each tube was placed into the spectrophotometer and the absorbance was taken.10. After 19 days, the results were analyzed using statistics and graphs.
1 3 5 7 9 12 15 190
0.1
0.2
0.3
0.4
0.5
0.6
0.7
Chlamydomonas Sample’s Absorbances
pH 7, 0x pH 7, 0.001x pH 7, .1xpH 5, 0x pH 5, 0.001x pH 5, .1x
Day of Experiment
Ab
so
rban
ce R
ead
ing
(n
m)
1 3 5 7 9 12 15 190
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
0.2Euglena Sample’s Absorbances
pH 7, 0x pH 7, 0.001x pH 7, .1xpH 5, 0x pH 5, 0.001x pH 5, .1x
Day of Experiment
Ab
sro
baan
ce R
ead
ing
(n
m)
pH 7, 0x pH 7, 0.001x
pH 7, 0.1x
pH 5, 0x pH 5, 0.001x
pH 5, 0.1x
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
Euglena Absorbance's: First & Last Days
Day1Day 19
Sample Group
Avera
ge A
bsorb
an
ce o
f S
am
ple
Gro
up
Statistical Analysis
Did the pH have an effect on the Chlamydomonas Samples at either 0x, 0.001x, or 0.1x fertilizer concentrations?
- Insignificant, the groups had p-values of >0.05 (p-values were 0.720 at 0x, 0.670 at 0.001x, and 0.249 at 0.1x ACCEPT NULL
Did the fertilizer concentrations have an effect on Chlamydomonas at a pH of 5 or 7?
- Significant, the p-value were: 7.714E-7 at pH7 & 3.37E-4 at pH5 REJECT NULL
Was there an interaction between pH and fertilizer concentration with the Chlamydomonas Samples?
- Insignificant, the p-value of 0.110516 is >0.05 ACCEPT NULL
ChlamydomonasANOVAS
Statistical Analysis Euglena
Did the pH have an effect on the Euglena Samples at either 0x, 0.001x, or 0.1x fertilizer concentrations?
-Significant at 0x (p-value of 0.002). REJECT NULL Insignificant at 0.001x or 0.1x because p-values were > than 0.05. (0.473 at 0.001x & 0.275 at 0.1x) ACCEPT NULL
Did the fertilizer concentrations have an effect on Euglena at a pH of 5 or 7?
-Insignificant, the p-values were 0.476 at pH7 & 0.333 at pH5 ACCEPT NULL
Was there an interaction between pH and fertilizer concentration with the Euglena Sample?
- Insignificant, the p-value of 0.429 is >0.05 ACCEPT NULL
ANOVAS
Dunnet’s TestFor the significant ANOVA’s, Dunnet’s tests were conducted to find which group(s) varied significantly from the control.
Chlamydomonas, pH7- The samples with a fertilizer concentration of 0.001x did not vary significantly from the control but the 0.1x samples did.
-T 2.53<3.03 T-crit [0.001x] T 10.22>4.63 T-crit [0.1x](very sure)
Chlamydomonas, pH5- The samples with a fertilizer concentration of 0.001x did not vary significantly from the control but the 0.1x samples did.
-T 1.616< 3.03 T-crit [0.001x] T 5.620>4.63 T-crit [0.1x] (very sure)
Euglena, 0x fertilizer- the pH difference was found to be significant.
-T 4.378> 4.03 T-crit (very sure)
CONCLUSIONSoThe null hypothesis is rejected because fertilizer concentration had an effect on Chlamydomonas and because pH did have an effect on Euglena at 0x. The null is accepted in that pH did not effect Chlamydomonas and fertilizer concentration had no effect on Euglena.
oResearch indicated Euglena would be effected by pH level. This was supported at a 0x fertilizer concentration.
oEutrophication may have played a role in Euglena’s spike and drop. It is possible it could have effected Chlamydomonas at a later time.
Notes/Variables/Limits
Algae shock?
Euglena Tube <pH 5, 0.001x>
Spectrometer accuracy
Cloudy and overcast days
Sample size limit
Could test thermal water pollution
Could test salt concentration
Use other organisms
EXTENSIONS
Bibliography
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