www.purdue.edu/ANE

Agriculture, Engineering, Science

Center for the Environment Birck Nanotechnology Center

Manufactured Nanomaterials in the Environment

Ron Turco, Loring Nies, Chad Jafvert, Bruce Applegate, Natalie Carroll,

Tim Filley, Robert Blanchette1, Leila Nyberg, Zhonghua Tong, Pradnya Kulkarni,

Marianne Bischoff and Benjamin Held1

Purdue University and University of Minnesota1

Acknowledgements The authors acknowledge support from

the National Science Foundation (NSF)

under Award EEC-0404006& United States EPA under Award RD-

83172001-0

RD-83172001-0

Nanotechnology Defined

• Working at length scales of ~1–100 nanometers [nm]– Results in new properties and functions

• Allows an ability to control (to see, measure, and manipulate) matter on the atomic & molecular scale

• Allows us to manipulate systems spanning from nano- to macroscopic scales

Start with a centimeter

Divide it into 10 equal parts (millimeter long)

Divide that into 10 equal parts (100 micrometers)

Divide that into 100 equal parts (micrometer)

Divide that into 10 equal parts (100 nanometers)

Divide that into 100 equal parts (Nanometer)

bean

flea

human hair

bacterium

virus

Nano- material

How small is a Nanometer?

1 cm

1 mm

100 m

1 m

100 nm

1 nm

1 cm = 10,000,000 nm

“There’s plenty of room at the bottom” (1959)

• The inspiration for nanotechnology came from Richard P. Feynman, 1959:

• ”The problems in chemistry and biology can be greatly helped if our ability to see what we are doing, and do thing on an atomic level, is ultimately developed – a development which I think cannot be avoided.”

Science (1985)

• Fullerenes are discovered by Robert Curl, Harold Kroto and Richard Smalley (Noble Prize)

• Spherical fullerenes -- buckyballs,

• Cylindrical nanotubes -- buckytubes

Nano-technology ~ 30 years later

• Eric Drexler (1986) Student in Feynman’s lab

Engines of Creation The Coming Era of Nanotechnology

(concept of “molecular manufacturing”)

Creation of the National Nanotechnology Initiative (NNI-Funding)(~25% DOD)

National Nanotechnology Initiative (NNI)

Budget History

The manufacturing technology of the 21st century

• Fabrication of devices with atomic or molecular scale precision

• Devices with some minimum feature sizes of less than 100 nanometers (nm) are considered to be products of nanotechnology

• The products can have quantum level features

From the Bottom• NT allows a bottom-up manufacturing

approach

• Nanotechnology processes can add:– material until the product has been created– makes the process similar to biological

systems– theoretically possible to start with one atom

• Production at 120 to 180 nm

Materials

• Lighter, stronger and programmable materials

• Lower failure rates and reduced life-cycle costs

• Better electrical efficiencies • Bio-inspired /Bio-reactive materials• Multifunctional, adaptive materials • Self-healing materials• Self-cleaning surfaces (e.g., windows)

Nano-based Products

• Computing, Data Storage, Electronics– Organic light-emitting diodes (OLEDs)– Nanoscale transistors– LCDs, LEDs, MP3s, electronic ink displays,

thin film batteries, and flexible electronics

• Integrated nanosensors: – Collecting, processing and communicating

data with minimal size, weight, and power consumption

Clothing/Film Products

• Color-changing fabrics

• Breathable waterproof ski jacket

• Wrinkle-resistant, stain-repellent threads

• Nanofilms are used now on eyeglasses, computer displays, and cameras to protect or treat the surfaces.

Health and Medicine Products

• Bandages embedded with silver nanoparticles – kill bacteria

• Drug delivery via a patch• Thin films on implantations into the human

body (for example screws, joints, and stents) allowing devices to last longer

• Respiration monitors that are many times more sensitive

• Nano-skin for skin graft applications

Health

• Nanocrystalline Sunscreen– Zinc oxide provides broad-spectrum

protection against UVA and UVB rays– Main ingredient is Z-COTE– Made with nanotechnology . . . Nano-

dispersed zinc oxide. . . . Goes from a white color to clear

Energy

• Energy Production: Clean, less expensive sources– Solar energy: Photovoltaic cells– Better Energy Utilization– Materials of construction sensing

changing conditions and in response, altering their inner structure

Environmental

• Environmental cleanup– Filters built out of carbon nanotubes, – Antifouling filters – Sieves that can filter bacteria and

poliovirus particles out of drinking water

Sports Products

• Tennis rackets – Nanotube Power and VS Nanotube Drive lightweight

• Tennis balls– Wilson Double Core tennis balls

• NanoDynamics golf ball – This ball is engineered with nanoparticles to spin less to

reduce the slice/hook problems

• Stronger golf clubs

• More accurate bowling balls

Potential Military NT(DARPA)

• Rugged/Embedded/Interlinked low-energy nanosensors to create pervasive networks

• Monitor for chemical & biological agents

• Implanted sensors for identification and health

Potential Military NT(DARPA)

• Nanofiber composites– Heat resistant, lighter and stronger– Cloaking devices

– Fuel Cells (H2 –storage) electric vehicles

• Strengthening of light armor• Better autonomous vehicles

– (combination of small electronics and nanofiber composites)

Potential Military NT(DARPA)

• Propellants and explosives with higher energy density

• Miniaturized guidance systems

Everything is great?

• Fears of Gray Goo

• Fears from “Prey”

• Comparisons of nano to biotech

• Generally no knowledge of environmental fate

• No data to back claims on either side of the argument

Exposure Routes

• Nanomaterials in clothing (uniforms) and equipment break off and enter the body and environment

• Nanoparticles as surface coverings erode and enter environment

• Nano-based fuels/explosives/ cloaking agents create residuals

A Nanomaterials Fate?

Introduction

Volatilization

Runoff

Leaching

Drainage -- Tile Flow

Degradation

DriftUptake

Sorption

Question: Is C60 is impacting the microbiology in the soil food web?

26

http://www.blm.gov/nstc/soil/bacteria/index.html

The talk presents the findings from a number of ongoing projects

27

Soils Work

Biosolids Work

Fungal Work

Typical Midwest Soils and chemical C60 preparations methods are established.

28

Texture

Name OM Sand Silt Clay

% pH %

Drummer 3.6 6 17 52 31

Tracy 1.5 5.5 55 37 8

Formation: Deguchi, et al., 2001

Concentration: Fortner et al., 2005

Size: DLS system

Our chosen soil microbiology methods are well established and documented

29

Microbial Form (PLFA/PCR-DGGE)Three domains model

Functions (CO2 CH4)

Size (Biomass)

Glucose Assimilation (14C-CO2)

Fungal Abilities (13C)

Evaluate Microbial Systems

C60 and nC60 had little impact on soil functions

Soil Respiration Biomass Size

30

Time (Days)

0 5 10 15 20 25 30 35

Cum

ulat

ive

CO

2 (

mg

CO

2 g

-1 so

il)

0.0

0.5

1.0

1.5

2.0

2.5

Soil THF nC60

C60

nC60 1 ppm / C60 1000 ppm – Drummer Soil

6-months

Soil-C THF-C C60 nC60

0

5

10

15

20

25

30

35

PL

PO

4 n

mo

l g-1 d

ry s

oil

No impact from longer incubations – Glucose assimilation testing method establishedTest procedure Response

31

Soil Challenged Soil Incubated Soil Tested

NanoMaterial

Time

Soil

14C-Glucose

14C-CO20 10 20 30 40 50 60 70

0

5

10

15

20

25

14C

O2

prod

uctio

n (%

of a

pplie

d 14

C-g

luco

se)

Time (h)

Soil-C THF-C C

60

nC60

Microbial profiling showed no difference after six months

32

M M Soil-C THF-C C60 nC60

Principal Componet 1( 58% of total variance)

-4 -3 -2 -1 0 1 2 3

Princi

pal C

om

ponent

2 (

19%

of

tota

l variance

)

-1.0

-0.5

0.0

0.5

1.0

1.5

2.0

C60

Soil

THFnC60

Fig. B

Principal Componet 1 (53% of total variance)

-4 -3 -2 -1 0 1 2 3

Princi

pal C

om

ponet

2 (

21%

of

tota

l variance

)

-1.0

-0.5

0.0

0.5

1.0

1.5

2.0

C60

Soil

nC60

THF

Fig. A

DGEE – 6 months PLFA 3 or 6 Months

Combinations of fullerenes with soil water stress show no effects

Five water potentials

Two nano materials

(nC60, C60, C12)

Two Soils

Respiratory response

33

Microbial activity measured as14CO2 evolved from 14C-glucose

treated soil exposed to nC60.

0 10 20 30 40

Cum

ula

tive 14 C

O2

evo

lve

d

(norm

aliz

ed

to c

on

trols

)

0

20

40

60

80

100

0.01 MPa0.03 MPa0.1 MPa0.5 MPa1.5 MPa

Drummer soil

Time (days)

0 10 20 30 40 50

Cum

ula

tive 14 C

O2

evo

lve

d

(norm

aliz

ed

to c

on

trols

)

0

20

40

60

80

100

Tracy soil

Soil diversity showed effects from C60 combined with water potential

Fatty Acids patterns from soils with nanomatrials and under water stresses (each symbol has an associated water potential)

34

PCA developed from FAMEs for treated and untreated soil.PC-1

-0.3 -0.2 -0.1 0.0 0.1 0.2 0.3

PC

-2

-0.3

-0.2

-0.1

0.0

0.1

0.2

0.3

Tracy Soil

Drummer Soil

Drummer nC60 controlDrummer nC60 treatedDrummer C60 controlDrummer C60 treatedDrummer C12 controlDrummer C12 treated

Tracy nC60 controlTracy nC60 treatedTracy C60 controlTracy C60 treatedTracy C12 controlTracy C12 treated

Preliminary data suggests nC60 crystal size had no effect on soil response

40 50 60 70 80 90 100 1100

2

4

6

8

10

12

14

16

18

Fre

qu

en

cy (

%)

Particle Diameter (nm)

Average Size 51 nm

0.00

5.00

10.00

15.00

20.00

25.00

30.00

35.00

40.00

Soil nC60-250nm

nC60-108nm

nC60-78nm

nC60-51nm

THF-C

% o

f ap

plie

d 14C

-glu

cose

0.00

5.00

10.00

15.00

20.00

25.00

30.00

Soil nC60-250nm

nC60-108nm

nC60-78nm

nC60-51nm

THF-C

% o

f ap

plie

d 14 C

-glu

cose

nC60 formed in different size classes (mixing speed) added to soil

Respiratory response

after 30 day exposure

Assessing the Impact of Nanomaterials on Anaerobic

Microbial Communities

Environmental Receptor: The Wastewater Treatment Plant

http://www.waterencyclopedia.com/images/wsci_04_img0570.jpg

Objectives and Hypotheses

• Objective I.

• Examine the effect of C60 on an anaerobic community.

• Hypothesis I. – C60 will remain inert and exert no detectable

toxic effects on anaerobic communities. Methanogenesis (community function) will be unaffected by treatment with C60.

Objectives and Hypotheses

• Objective II. • Develop Three-Domain Community

Analysis

• Hypothesis II. – Three-Domain Community Analysis will detect

shifts in anaerobic communities more completely than the more widely used analysis of community structure in a single domain.

Community Function Measured by Anaerobic Toxicity Assay

+/- Substrate:

C6H12O6

CH3OH

CH3CH2OH

(G/M/E)

Concentration – Dependent Antibiotic Toxicity

0

20

40

60

80

100

120

140

160

0 10 20 30 40 50 60 70 80

Time (days)

Vo

lum

e (

mL

)

- G/M/E reference. - 2 mg/L metronidazole + G/M/E. - 200 mg/L metronidazole + G/M/E.

C60 Did Not Inhibit Gas Formation

0

100

200

300

400

500

600

0 50 100 150

time (days)

Ga

s V

olu

me

(m

l)

- G/M/E Substrate Reference

- C60 dissolved in MeOH/EtOH + G/M/E

▬ - aqeuous suspension C60 + G/M/E

▲- C60 dissolved in toluene, plated on dried sludge + G/M/E

Antibiotic treatment induced communi

ty chagnes shifts in all three domains R1 M1 R2 M2

R1 m M1

R1 R2 R2*M1 M2 M2*

Archaea Eukarya Bacteria

C60 did not affect Archaeal Community Profile

A B C H1 D F G H2

Increasing % G+C

Denaturing gradient gel electrophoresis with Archaea primers, showing similar community profiles at the end of each experiment for treated samples and reference samples.

No Bacterial

Community Shifts with C60

treatment

A B C H1 D F G H2

Increasing % G+C

DGGE with Bacteria primers, showing similar community profiles at the end of each experiment for treated samples and reference samples.

Conclusions and Significance

• No evidence of C60 toxicity to any subset of the microbial community, No evidence of major community shifts

• No methods for measurement of nanomaterials or products in the environment, nano risk assessment not yet standardized

• Important role for analysis of microbial community structure and function

• Long-term studies of C60 in the environment will be necessary to determine biodegradation potential

Outreach

• Mission (objectives) of the outreach program are helping the general public, especially high school students, understand the science behind the manufactured nanoparticles.

• www.purdue.edu/ANE

Outreach

www.purdue.edu/ANE