Red Acid Dye Effects of Microbial Life

Ryan KramerGrade 11

Pittsburgh Central Catholic

Problem

• Water systems are polluted with:• Antibiotics• Pharmaceuticals (Estrogen, etc.)• Mercury• Sulfur• Sewage• Fertilizers• Organics• Cleaning projects• Dyes and other colored agents

Dye Effects on Water

• Studies have shown that dyes add unwanted colors and toxins to water that effect microbial life and algal photosynthetic rates.

Euglena Gracilis

• Unicellular organism from the Protista Kingdom.

• Both autotrophic and heterotrophic.• Found in fresh and salt water.• Reproduce rapidly.• Absorbs light readily.

Acid Dye

• Used by companies to color clothing, textiles, ect.• Researchers claim they’re non toxic and have no

harmful effect on aquatic life.• Billions of gallons dumped changed photosynthetic

rates and alter PH harming aquatic life.• Modern chemistry still can’t prevent all dye

problems• Sudan I crisis in the UK when a carcinogenic dye got in over 400 super market items.

Food Coloring

• Studies have shown colors affect taste• For this reason companies add colors to enhance dining

experience

• It is a non-toxic dye.• Chosen because

• Its non-toxic• Easy to obtain• Will test if color alone will effect algal growth

Spectrophotometer

• Device measuring light intensity or wavelengths of light

• Used for measuring light absorption• Measures the light absorption of the algae

• More absorption means more algae• Wavelength: 430nm

Purpose• To determine if dyes have an

effect on the growth of the Euglena Gracilis.

Hypothesis

• Null Hypotheses: Adding color will not have a significant effect on algal growth.

• Alternative Hypotheses: Adding color

will have a significant effect on algal growth.

Materials

• Science Kit Educator Spectrophotometer

• 80 13x100 mm borosilicate culture test tubes

• Red food coloring• Micropipette• Test tube rack• Pipette tips

• Red Acid Dye• Euglena Gracilis• Microscope• Hemocytometer• Spring Water • Soil Water

Procedure

• Stock solutions of food coloring and dye were generated.

Spring H2O Soil H2O Euglena AD/FC

0% 2mL 1mL 2mL 0mL

1% 1.90mL 1mL 2mL 10uM

5% 1.50mL 1mL 2mL 50uM

Procedure

• The spectrophotometer was blanked and absorbance readings were taken.

• Absorbance readings for each tube were taken at 430 nm every day for two weeks.

• After two weeks a cell count was taken with a hemocytometer at 100 times magnification to check if the cells were alive after 2 weeks.

Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 141.2

1.25

1.3

1.35

1.4

1.45

1.5

1.55

1.6

Control1% FC5% FC1% AD5% AD

Extent of Time

Abs.

at 4

30 n

m

Euglena Absorbance

P Value: 6.064x10E-29

Dunnett’s Test

Variable Group T Value Interpretation

1 % FC 3.51 Significant

5 % FC 5.43 Significant

1% AD 5.51 Significant

5% AD 10.63 Significant

T Crit: 3.48

Euglena Cell Counts After 14 Days

P Value: 4.43E-12

Control 1% FC 5% FC 1% AD 5% AD0

5000

10000

15000

20000

25000

30000

35000

40000

45000

Variable Group

Num

ber o

f Cel

ls

Dunnett’s Test

Variable Group T Value Interpretation

1 % FC 3.57 Significant

5 % FC 5.289 Significant

1% AD 6.289 Significant

5% AD 11.889 Significant

T Crit: 3.48

Conclusion

• The null hypothesis was rejected for all four groups in the absorbance and cell count tests.

• This means that the algae survivorship was significantly effected by the red colored dyes.

• The most likely explanation for the decline in algal growth was the added color effected photosynthetic rates.

Limits and Extension

• Limits• The test tubes may have receive differing amounts of

sun light which could change photosynthesis.• Only one type of algae was used in this experiment.

• Extensions• Test more colors of dye.• Introduce a compound to break down the dye to see if

dye break down affect the algal growth.

References • K.Suthagar. POLLUTION_CONTROL_IN_DYE_INDUSTRY-_22-04-09.

National Centre for Catalysis Research. PPT.• "Water Pollution." MBGnet. Web. 05 Nov. 2011.

<http://www.mbgnet.net/fresh/pollute.htm>.• "BBC NEWS | Health | Food Recalled in Cancer Dye Scare." BBC News

- Home. Web. 05 Nov. 2011. <http://news.bbc.co.uk/2/hi/health/4277677.stm>.

• "Acid Dye." Wikipedia, the Free Encyclopedia. Web. 05 Nov. 2011. <http://en.wikipedia.org/wiki/Acid_dye>.

• "Food Coloring." Wikipedia, the Free Encyclopedia. Web. 06 Nov. 2011. <http://en.wikipedia.org/wiki/Food_coloring>.

• http://www.emc.maricopa.edu/faculty/farabee/biobk/biobookps.html