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

Algal ImportanceBase of aquatic food chainUsed as a bio-indicator for aquatic environments

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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