Effect of Probiotic Tibial Weight In Male Broiler Chickens

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www.purdue.edu/ANE
Agriculture, Engineering, Science Center for
the Environment Birck Nanotechnology Center
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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
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Acknowledgements

The authors acknowledge support from the
National Science Foundation (NSF) under Award
EEC-0404006 United States EPA under Award
RD-83172001-0
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Nanotechnology Defined

• Working at length scales of 1100 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

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How small is a Nanometer?
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
1 cm 10,000,000 nm
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Theres 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.

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Science (1985)

• Fullerenes are discovered by Robert Curl, Harold
  Kroto and Richard Smalley (Noble Prize)
• Spherical fullerenes -- buckyballs,
• Cylindrical nanotubes -- buckytubes

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Nano-technology 30 years later

• Eric Drexler (1986) Student in Feynmans lab
• Engines of Creation
• The Coming Era of Nanotechnology
• (concept of molecular manufacturing)

Creation of the National Nanotechnology
Initiative (NNI-Funding) (25 DOD)
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National Nanotechnology Initiative (NNI) Budget
History
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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

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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

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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)

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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

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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.

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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

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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

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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

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Environmental

• Environmental cleanup
• Filters built out of carbon nanotubes,
• Antifouling filters
• Sieves that can filter bacteria and poliovirus
  particles out of drinking water

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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

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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

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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)

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Potential Military NT(DARPA)

• Propellants and explosives with higher energy
  density
• Miniaturized guidance systems

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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

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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

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A Nanomaterials Fate?
Drift
Uptake
Introduction
Volatilization
Sorption
Degradation
Runoff
Drainage -- Tile Flow
Leaching
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Question Is C60 is impacting the microbiology
in the soil food web?
http//www.blm.gov/nstc/soil/bacteria/index.html
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The talk presents the findings from a number of
ongoing projects
Soils Work
Biosolids Work
Fungal Work
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Typical Midwest Soils and chemical C60
preparations methods are established.
Formation Deguchi, et al., 2001 Concentration
Fortner et al., 2005 Size DLS system
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Our chosen soil microbiology methods are well
established and documented

• Evaluate Microbial Systems

Microbial Form (PLFA/PCR-DGGE) Three domains
model Functions (CO2 CH4) Size
(Biomass) Glucose Assimilation (14C-CO2) Fungal
Abilities (13C)
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C60 and nC60 had little impact on soil functions

• Soil Respiration

• Biomass Size

6-months
nC60 1 ppm / C60 1000 ppm Drummer Soil
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No impact from longer incubations Glucose
assimilation testing method established

• Test procedure

• Response

Soil Challenged Soil Incubated Soil
Tested
NanoMaterial
Soil
14C-CO2
Time
14C-Glucose
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Microbial profiling showed no difference after
six months

• PLFA 3 or 6 Months

• DGEE 6 months

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Combinations of fullerenes with soil water stress
show no effects

• Five water potentials
• Two nano materials
• (nC60, C60, C12)
• Two Soils
• Respiratory response

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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)

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Preliminary data suggests nC60 crystal size had
no effect on soil response
nC60 formed in different size classes (mixing
speed) added to soil Respiratory response
after 30 day exposure
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Assessing the Impact of Nanomaterials on
Anaerobic Microbial Communities
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Environmental Receptor The Wastewater Treatment
Plant
http//www.waterencyclopedia.com/images/wsci_04_im
g0570.jpg
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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.

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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.

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Community Function Measured by Anaerobic Toxicity
Assay
/- Substrate C6H12O6 CH3OH CH3CH2OH (G/M/E)
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Concentration Dependent Antibiotic Toxicity

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

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C60 Did Not Inhibit Gas Formation

• - 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

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Antibiotic treatment induced community chagnes
shifts in all three domains
Archaea
Eukarya
Bacteria
R1 m M1
M1 M2 M2
R1 M1 R2 M2
R1 R2 R2
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C60 did not affect Archaeal Community
Profile
Increasing GC
A B C H1 D F G H2
Denaturing gradient gel electrophoresis with
Archaea primers, showing similar community
profiles at the end of each experiment for
treated samples and reference samples.
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No Bacterial Community Shifts with C60
treatment
Increasing GC
A B C H1 D F G H2
DGGE with Bacteria primers, showing similar
community profiles at the end of each experiment
for treated samples and reference samples.
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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

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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

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Outreach
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www.purdue.edu/ANE