Soil Contamination Effects on Microbial Life

Christian Dresser- Pittsburgh Central Catholic High School Grade 10

Common Inorganic Soil Contaminants

Lead - poisonous paint can gradually come off buildings and contaminant soils

Mercury ndash poisonous generated from cement production coal fired plants etc

Acetone ndash used in fuel for cars and trains dissipates slowly in soil Arsenic ndash poisonous used in integrated circuits Barium ndash poisonous used in welding railroad tracks together Benzene ndash produced from coal found in exhaust Coal - carried via near by trains combustible Gypsum ndash used in Ordinary Portland Cement (OPC) drywall and

plaster composed of calcium sulfate

Soils

Soils are among the most precious though least appreciated resources for human kind

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

The world is facing a crisis of soil integrity erosion

salinization compaction Pollution

Soils

Soils near industrial urban or transportation centers are considered at high risk for soil pollution

Organic ndash bacteria viruses molds etc Inorganic ndash lead mercury acetone arsenic barium

benzene coal and gypsum

Lead Poisonous can cause blood and brain disorders Highly malleable and ductile 8 million tones produced annually Used in car batteries organ pipes and stained glass

windows Lead paint gradually sheds off buildings and contaminants

soils

Mercury Used in thermometers barometers and other scientific

apparatuses Poisonous can cause brain damage can be absorbed and

inhaled through the skin and mucous membranes Generated from cement production

Acetone Component of paints and varnishes Used to dissolve many plastics Solvent can depress the central nervous system Dissipates slowly in soil Used in fuel for cars and trains

Arsenic Poisonous metalloid with many allotropic forms Used in pesticides herbicides insecticides and various

alloys Used in integrated circuits Exposure to high level can cause multi-system organ failure

Coal Carrieddelivered via near by trains Largest source of fuel for the generation of

electricity World consumption use is 62 billion tons annually Used to produce syngas a mixture of carbon

monoxide (CO) and hydrogen gas (H2) Combustible sedimentary rock

Barium Used in rat poison making bricks and glass

making Extremely poisonous effects the nervous

system Used in welding rail tracks together

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Common Inorganic Soil Contaminants

Lead - poisonous paint can gradually come off buildings and contaminant soils

Mercury ndash poisonous generated from cement production coal fired plants etc

Acetone ndash used in fuel for cars and trains dissipates slowly in soil Arsenic ndash poisonous used in integrated circuits Barium ndash poisonous used in welding railroad tracks together Benzene ndash produced from coal found in exhaust Coal - carried via near by trains combustible Gypsum ndash used in Ordinary Portland Cement (OPC) drywall and

plaster composed of calcium sulfate

Soils

Soils are among the most precious though least appreciated resources for human kind

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

The world is facing a crisis of soil integrity erosion

salinization compaction Pollution

Soils

Soils near industrial urban or transportation centers are considered at high risk for soil pollution

Organic ndash bacteria viruses molds etc Inorganic ndash lead mercury acetone arsenic barium

benzene coal and gypsum

Lead Poisonous can cause blood and brain disorders Highly malleable and ductile 8 million tones produced annually Used in car batteries organ pipes and stained glass

windows Lead paint gradually sheds off buildings and contaminants

soils

Mercury Used in thermometers barometers and other scientific

apparatuses Poisonous can cause brain damage can be absorbed and

inhaled through the skin and mucous membranes Generated from cement production

Acetone Component of paints and varnishes Used to dissolve many plastics Solvent can depress the central nervous system Dissipates slowly in soil Used in fuel for cars and trains

Arsenic Poisonous metalloid with many allotropic forms Used in pesticides herbicides insecticides and various

alloys Used in integrated circuits Exposure to high level can cause multi-system organ failure

Coal Carrieddelivered via near by trains Largest source of fuel for the generation of

electricity World consumption use is 62 billion tons annually Used to produce syngas a mixture of carbon

monoxide (CO) and hydrogen gas (H2) Combustible sedimentary rock

Barium Used in rat poison making bricks and glass

making Extremely poisonous effects the nervous

system Used in welding rail tracks together

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Soils

Soils are among the most precious though least appreciated resources for human kind

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

The world is facing a crisis of soil integrity erosion

salinization compaction Pollution

Soils

Soils near industrial urban or transportation centers are considered at high risk for soil pollution

Organic ndash bacteria viruses molds etc Inorganic ndash lead mercury acetone arsenic barium

benzene coal and gypsum

Lead Poisonous can cause blood and brain disorders Highly malleable and ductile 8 million tones produced annually Used in car batteries organ pipes and stained glass

windows Lead paint gradually sheds off buildings and contaminants

soils

Mercury Used in thermometers barometers and other scientific

apparatuses Poisonous can cause brain damage can be absorbed and

inhaled through the skin and mucous membranes Generated from cement production

Acetone Component of paints and varnishes Used to dissolve many plastics Solvent can depress the central nervous system Dissipates slowly in soil Used in fuel for cars and trains

Arsenic Poisonous metalloid with many allotropic forms Used in pesticides herbicides insecticides and various

alloys Used in integrated circuits Exposure to high level can cause multi-system organ failure

Coal Carrieddelivered via near by trains Largest source of fuel for the generation of

electricity World consumption use is 62 billion tons annually Used to produce syngas a mixture of carbon

monoxide (CO) and hydrogen gas (H2) Combustible sedimentary rock

Barium Used in rat poison making bricks and glass

making Extremely poisonous effects the nervous

system Used in welding rail tracks together

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Soils

Soils near industrial urban or transportation centers are considered at high risk for soil pollution

Organic ndash bacteria viruses molds etc Inorganic ndash lead mercury acetone arsenic barium

benzene coal and gypsum

Lead Poisonous can cause blood and brain disorders Highly malleable and ductile 8 million tones produced annually Used in car batteries organ pipes and stained glass

windows Lead paint gradually sheds off buildings and contaminants

soils

Mercury Used in thermometers barometers and other scientific

apparatuses Poisonous can cause brain damage can be absorbed and

inhaled through the skin and mucous membranes Generated from cement production

Acetone Component of paints and varnishes Used to dissolve many plastics Solvent can depress the central nervous system Dissipates slowly in soil Used in fuel for cars and trains

Arsenic Poisonous metalloid with many allotropic forms Used in pesticides herbicides insecticides and various

alloys Used in integrated circuits Exposure to high level can cause multi-system organ failure

Coal Carrieddelivered via near by trains Largest source of fuel for the generation of

electricity World consumption use is 62 billion tons annually Used to produce syngas a mixture of carbon

monoxide (CO) and hydrogen gas (H2) Combustible sedimentary rock

Barium Used in rat poison making bricks and glass

making Extremely poisonous effects the nervous

system Used in welding rail tracks together

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Lead Poisonous can cause blood and brain disorders Highly malleable and ductile 8 million tones produced annually Used in car batteries organ pipes and stained glass

windows Lead paint gradually sheds off buildings and contaminants

soils

Mercury Used in thermometers barometers and other scientific

apparatuses Poisonous can cause brain damage can be absorbed and

inhaled through the skin and mucous membranes Generated from cement production

Acetone Component of paints and varnishes Used to dissolve many plastics Solvent can depress the central nervous system Dissipates slowly in soil Used in fuel for cars and trains

Arsenic Poisonous metalloid with many allotropic forms Used in pesticides herbicides insecticides and various

alloys Used in integrated circuits Exposure to high level can cause multi-system organ failure

Coal Carrieddelivered via near by trains Largest source of fuel for the generation of

electricity World consumption use is 62 billion tons annually Used to produce syngas a mixture of carbon

monoxide (CO) and hydrogen gas (H2) Combustible sedimentary rock

Barium Used in rat poison making bricks and glass

making Extremely poisonous effects the nervous

system Used in welding rail tracks together

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Mercury Used in thermometers barometers and other scientific

apparatuses Poisonous can cause brain damage can be absorbed and

inhaled through the skin and mucous membranes Generated from cement production

Acetone Component of paints and varnishes Used to dissolve many plastics Solvent can depress the central nervous system Dissipates slowly in soil Used in fuel for cars and trains

Arsenic Poisonous metalloid with many allotropic forms Used in pesticides herbicides insecticides and various

alloys Used in integrated circuits Exposure to high level can cause multi-system organ failure

Coal Carrieddelivered via near by trains Largest source of fuel for the generation of

electricity World consumption use is 62 billion tons annually Used to produce syngas a mixture of carbon

monoxide (CO) and hydrogen gas (H2) Combustible sedimentary rock

Barium Used in rat poison making bricks and glass

making Extremely poisonous effects the nervous

system Used in welding rail tracks together

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Acetone Component of paints and varnishes Used to dissolve many plastics Solvent can depress the central nervous system Dissipates slowly in soil Used in fuel for cars and trains

Arsenic Poisonous metalloid with many allotropic forms Used in pesticides herbicides insecticides and various

alloys Used in integrated circuits Exposure to high level can cause multi-system organ failure

Coal Carrieddelivered via near by trains Largest source of fuel for the generation of

electricity World consumption use is 62 billion tons annually Used to produce syngas a mixture of carbon

monoxide (CO) and hydrogen gas (H2) Combustible sedimentary rock

Barium Used in rat poison making bricks and glass

making Extremely poisonous effects the nervous

system Used in welding rail tracks together

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Arsenic Poisonous metalloid with many allotropic forms Used in pesticides herbicides insecticides and various

alloys Used in integrated circuits Exposure to high level can cause multi-system organ failure

Coal Carrieddelivered via near by trains Largest source of fuel for the generation of

electricity World consumption use is 62 billion tons annually Used to produce syngas a mixture of carbon

monoxide (CO) and hydrogen gas (H2) Combustible sedimentary rock

Barium Used in rat poison making bricks and glass

making Extremely poisonous effects the nervous

system Used in welding rail tracks together

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Coal Carrieddelivered via near by trains Largest source of fuel for the generation of

electricity World consumption use is 62 billion tons annually Used to produce syngas a mixture of carbon

monoxide (CO) and hydrogen gas (H2) Combustible sedimentary rock

Barium Used in rat poison making bricks and glass

making Extremely poisonous effects the nervous

system Used in welding rail tracks together

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Barium Used in rat poison making bricks and glass

making Extremely poisonous effects the nervous

system Used in welding rail tracks together

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Benzene

Component of cigarette smoke and found in exhaust

Produced from coal Used to make polymers and plastic Serious health effects including multi -

organ cancer and even death

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Experimental Soil Samples Industrial Soil ndash located near trains and a cement

factory possibly exposed to the inorganic contaminants such as lead mercury and gypsum

Non-Industrial Soil ndash not located near trains nor a cement factory most likely not exposed to inorganic contaminants

Control Soil ndash store-bought potting soil not exposed to any inorganic contaminants

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Cement

Made by heating limestone with small quantities of other materials (such as clay) to 1450deg C

The resulting hard substance called clinker is ground with a small amount of gypsum into a powder to make Ordinary Portland Cement (OPC)

Basic ingredient of concrete mortar and grout

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Soil

Broken rock particles that have been altered by chemical and environmental conditions weathering and erosion

Altered by interactions between the lithosphere hydrosphere atmosphere and biosphere

Potting soil contains peat moss composted bark sand and perlite

It also contains vermicompost for water retention

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Gypsum Composed of calcium sulfate CaSO4 2H2O Occurs as twinned crystals and transparent

cleavable masses called selenite Deposited in lakes oceans and hot springs Used in drywall plaster soil conditioner fertilizer

blackboard chalk and OPC

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

E Coli Escherichia coli (E coli) ndash very common found in

intestinal tract of most mammals There are many strains of E coli most are non-

pathogenic Pathogenic strains can cause illness and death in

humans Frequently studied in biology ndash ubiquitous simple

structure easily manipulated in the laboratory

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Purpose

The purpose of this investigation was to determine the effects of soil componentscontaminants on the survivorship of Ecoli

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Null Hypothesis

The addition of soil to Ecoli suspensions will not significantly affect survivorship

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Materials 500 grams of sterilized soil from an industrial site (railroad tracks and

cement factory) 500 grams of sterilized soil from a non-industrial site 500 grams of sterilized control soil (store bought) Sterile Dilution Fluid (SDF) (10mM KH2PO4 10mM K2HPO4 1mM MgSO4

01mM CaCl2 100mM NaCl) 3 tubes for the 3 different soil extracts ( 300mL SDF and 500 g soil for

each tube) 14 tubes for the low and high concentrations of the extracts (Low- 89mL

SDF 10mL extract High ndash 99mL extract) Ecoli bacteria (1 mL in each of the 14 tubes (107 cellsmL) ) 56 LB Agar plates ( 8 for each concentration) Sterile Pipettes LB media (yeast extract (05 ) NaCl (1) tryptone (15)) Bunsen burner Ethanol Autoclave Klett Spectrophotometer

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Procedure 1 E Coli was grown overnight in sterile media 2 A sample of the overnight culture was added to fresh

media in a sterile sidearm flask 3 The culture was placed in a shaking water bath until a

density of 50 Klett spectrophotometer units was reached This represents a cell density of approximately 108 cellsmL

4 The culture was diluted in SDF to a concentration of approximately 103 cellsmL

5 Soils from 3 different sites were sterilized in an autoclave for 45 minutes

6 500 g of each sterilized type of soil were weighed 7 500 g of each type of soil were added to 300mL of SDF

creating a soil extract

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Procedure - ConcentrationsLow

ConcentrationIndustrial Non-

IndustrialControl Soil

SDF 89mL 89mL 89mLExtract 10mL 10mL 10mLEcoli 01mL 01mL 01mL

High Concentration

Industrial Non-Industrial Control Soil

SDF 00mL 00mL 00mLExtract 99mL 99mL 99mLEcoli 01mL 01mL 01mL

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Procedure Contrsquod

11 01mL of the E Coli cell culture was added to each test tube yielding a final volume of 100mL and a cell density of approximately 107 cells mL

12 The solution in each tube was mixed by vortexing and allowed to sit at room temperature for 15 minutes

13 After vortexing to evenly suspended cells 01mL aliquots were removed from the tubes and spread on LB-Agar plates

14 The plates were incubated at 37 degrees Celsius overnight

15 The resulted colonies were counted Each colony is assumed to have arisen from 1 cell

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Data Ave Small Ave Large Ave Total

Non ndash Industrial Low

24 31 55

Non ndash Industrial High

11 9 20

Industrial Low 15 16 32Industrial High 21 16 37

Control Dirt Low

8 13 22

Control Dirt High

22 14 36

Control SDF 16 14 29

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Total Average of Colonies Counted

plt05

plt05

plt05

plt05

plt05plt05

0

10

20

30

40

50

60

Soils and Concentrations

Col

onie

s C

ount

edP-value = 0000184 plt05 = Significant

plt 05

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Dunnet Test Results Variables t value Interpretation

Control Dirt Higha = 05 n = 8 a = 01

12227 Not SignificantNot Significant

Control Dirt Lowa = 05 n = 65 a

= 01

10417 Not SignificantNot Significant

Industrial Higha = 05 n = 7 a =01

13344 Not SignificantNot Significant

Industrial Lowa = 05 n = 8 a = 01

23666 Not SignificantNot Significant

Non-Industrial Higha = 05 n = 75 a

= 01

13504 Not SignificantNot Significant

Non-Industrial Lowa = 05 n = 75 a

= 01

39529 SignificantNot Significant

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Conclusion

In my hypothesis I stated that the E coli grown in the simple SDF fluid would have the greatest survival rate

Through the experiment this was proven wrong

The E Coli grown in a low concentration of the non-industrial soil had the greatest survival rate while the E Coli grown in the simple SDF solution (control) had the fifth highest survival rate

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Conclusion

Through the experiment there was a Rejection of the Null hypothesis The soil contaminants did have effects on the survivorship of EColi

In comparison with the control SDF through a Dunnetrsquos test the survivorship of the EColi in the Non-Industrial Low was significant to the survivorship of the EColi in the control SDF

This was the only concentration that was significant which was proven in the Dunnetrsquos test

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

Future Implications More soils from different locations could be tested The soils could be tested on the survivorship of yeast More LB agar plates could be used for each type of soil Different concentrations of the soil extracts could be tested

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References

References Dr John Wilson biostatistician University of Pittsburgh httpenwikipediaorgwikiLead httpenwikipediaorgwikiMercury httpenwikipediaorgwikiAcetone httpenwikipediaorgwikiArsenic httpenwikipediaorgwikiBarium httpenwikipediaorgwikiBenzene httpenwikipediaorgwikiCoal httpenwikipediaorgwikiCement httpenwikipediaorgwikiGypsum httpenwikipediaorgwikiSoil httpenwikipediaorgwikiE Coli httpwwwepagovebtpagespollsoilcontaminantshtml httpwwwecywagovprogramshwtrdemodebrispages2dwinsoilhtml httpwwwegrmsuedu~envirotoolscgi-binsoilphp3

• Soil Contamination Effects on Microbial Life
• Common Inorganic Soil Contaminants
• Soils
• Soils (2)
• Lead
• Mercury
• Acetone
• Arsenic
• Coal
• Barium
• Benzene
• Experimental Soil Samples
• Cement
• Soil
• Gypsum
• E Coli
• Purpose
• Null Hypothesis
• Materials
• Procedure
• Procedure - Concentrations
• Procedure Contrsquod
• Data
• Slide 24
• Dunnet Test Results
• Conclusion
• Conclusion (2)
• Future Implications
• References