Nanotechnology holds promise for advances in biotechnology and medicine. Its applications could include building faster computers, more efficient solar cells, finding tumors using nanorobots, and improving agriculture. In animal reproduction, nanotechnology is still in its infancy but may transform the livestock sector. Examples include using nanoparticles for sperm preservation and imaging, microencapsulation of sperm for controlled release, and biosensors to detect reproductive hormones for estrus detection. While nanotechnology shows potential, oversight is needed to ensure safety and address societal issues as these applications develop further.

1. An Overview of
Applications
in Animal Reproduction

2. Science has advanced to the point that cutting edge
research involves working with individual atoms and
molecules
Nanotechnology holds the promise to exceed the advances
achieved in recent decades in biotechnology and medicine
Nanotechnologies are one of the most prominent actors of
the scientific revolution marking the beginning of the new
Millennium

3. Its applications are expected to have dramatic
impacts
building tremendously faster computers,
constructing lighter aircrafts,
finding tumors using nanorobots
generating vast amounts of energy from highly efficient solar cells
possibly improving agriculture and livestock sector
C The application in the animal sector is still in its
infancy & is predicted to transform the entire livestock
sector

5. • Nanotechnology
Branch of technology that deals with dimensions &
tolerances of 0.1-100 nm. It is the engineering of functional
systems at the molecular scale
Technology devoted to manipulation of atoms &
molecules leading to construction of structures in the
nanoscale size range retaining unique properties
Nanobiotechnology
Branch of nanotechnology with biological & bio-
chemical application or uses
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6. History…
First use of the concepts in "There's Plenty of Room at the Bottom," a
talk given by him at American Physical Society meeting at Caltech on
29,Dec ’59
Richard Feynman - The theoretical capability was
envisioned as early as 1959 by this renowned
physicist
Grand father of nanotechnology
K. Eric Drexler popularized the word
'nanotechnology' in the 1980's-building machines on
the scale of nanometers
Father of nanotechnology
R.Feynman
K.E.Dexler
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7.  Countries in the world are investing to secure a market
share
 US leads with a 4 yr, 3.7 billion US dollars investment
through its National Nanotechnology Initiative (NNI)
 China’s share of publications rose from 7.5% in ’95 to 18.3%
in ’04, taking that country from 5th to 2nd among the world
leaders
 potential of nanotechnology to revolutionize the health
care, textile, materials, information and communication
technology and energy sectors. Several products enabled
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10. Nanomaterials are structures created by nanotechnology
research that range from 1 to 100 nanometers in size
Lighter
Stronger
Faster
Smaller
Durable
Common examples are
fullerenes, nanotubes,
buckyballs, quatum dots,
dendrimers & nanoshells

11. ELECTRONIC
PHYSICAL SCIENCES
SPACE RESEARCH
FUEL CELLS-SOLAR CELLS
ENVIRONMENT PROTECTION- WATER, AIR, TOXICANT
DETECTOR
FABRICS
SPORTING GOODS
MEDICINE & REPRODUCTION...??
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12. Micro/nanofluidics
Biosensors
Micro/nano
encapsulation

14. Process in which tiny particles or droplets are surrounded
by a coating to give small capsules many useful properties
It is a small sphere with semi permeable membrane around it
Material inside is referred to as the core, internal phase, or
fill & the wall is sometimes called a shell, coating or
membrane
Sustained sperm release
timing of AI less critical
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15. Pan coating, Air-suspension coating, Centrifugal
extrusion, Vibrational Nozzle, polymerization. Done at 23° C
 Sodium alginate, high calcium buffer, barium alginate
 Cellulose sulfate (CS)-poly-diallyldimethyl ammonium
chloride -(pDADMAC)
 Poly-l-lysine, polyvinylamine and protamine sulfate

17. Encapsulation has been successful with capsules ranging
in size from 0.75 to 1.5 mm, and with sperm concentrations
from 45 to 180 x 10 (6) cells per mL
S-pDADMAC-based capsules break up within 72 h after
addition of either purified cellulase or cellulase-filled
alignate-Ca2+ capsules
Fertility studies indicate that sperm encapsulated with poly-
l-lysine or protamine sulfate may achieve normal fertility
Nebel et al., (1998)
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18.  sperms are highly susceptible to cold shock
 prolonged storage at ultra low temperatures reduces survival
rate
Alginate micro-encapsulation process can improve
 longevity of boar spermatozoa stored at 5°C &
 fertility of microencapsulated spermatozoa in vivo

19.  Novel applications of nanostructured silica in biotechnology
& reproduction
 Nanosize fumed silica, used as a carrier in nanomaterials,
proved to provide itself positive biological effect with respect
to reproductive cells particularly sperms
 New silica based nanomaterials as admixtures to existing
cryomedia were synthesized & successfully tested with post-
thaw bovine sperm & human reproductive cells to increase
their viability, longevity

21. Use of nanoparticles for imaging has gained
considerable momentum in recent years
Fluorescent Quantum Dots (QDs), which are small
nanocrystals (1–10 nm) made of inorganic semiconductor
materials, possess several unique optical properties best
suited for in vivo imaging
In addition, QDs are extremely photostable, making them
ideally suited for long-term observations or repeated
measurements
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22.  Nanoparticles consisting of a core of biodegradable PLGA
with free pendant chains of functionalized poly(ethylene
glycol) onto which the targeting moieties are conjugated
 Targeting moiety is a bioactive ligand, either FSH or LH,
that is specifically recognized & captured by the
corresponding bioactive receptors
 are highly expressed in ovaries, & causing nanoparticles
to localize in the ovaries prior to MRI imaging

23. Detection of cancer protein biochips consisting of micro- &
nanoarrays whereby quantum dots (QDs) conjugated to
antibodies (Abs) of prostate specific antigens (PSA) has been
used for detection of various clinical biomarkers
Potential offered by QDs in in vitro analysis of cancer
biomarker imaging Gokarna et al.,
(2008)

24. image shows prostate cancer cells that
have taken up fluorescently labeled
nanoparticles (shown in red)
Nuclei and cytoskeletons are stained blue and green resp.
Similarly designed targeted nanoparticles are capable of
getting inside cancer cells & releasing lethal doses of
chemotherapeutic drugs to eradicate tumors
Benjamin., (2006)

25. •
AFM is one of the foremost tools for imaging, measuring, &
manipulating matter at the nanoscale
Only technique capable of real-time imaging of the surface of
living cell at nano-resolution
Provides advantage of directly observing living biological
cells in their native environment
Sperm head defects & acrosome can be examined and
correlated with the lack of functional integrity of the cell

26. Unstained, unfixed sperms in their natural physiological
surroundi-ngs can be imaged which provides valuable details
on various defects in sperm cells as 3D images with precise
topographical details

28.  Nanotechnology recently has emerged as one of the most
propitious field in cancer treatment
 Targeted nanomedicines may aid in evading the adverse effects
(such as immunosuppression, cardiomyopathy, and neurotoxicity)

29. A nanoparticulate system to deliver a diphtheria toxin suicide
gene (DT-A) driven by a prostate specific promoter to cells
Using a degradable, poly(beta-amino ester) polymer, poly(butane
diol diacrylate co amino pentanol) (C32)
Nearly 50% of normal prostates showed a significant reduction in
size, attributable to cellular apoptosis, whereas injection with naked
DT-A-encoding DNA had little effect Peng et
al.,(2007)
Significant apoptosis was also observed in C32/DT-A injected
prostate tumors with no damage to surrounding tissue

30. Studies with nanoparticle-hypericin formulation showed that the
combination was more effective than hypericin itself at killing
ovarian cancer cells
FSHR can be targeted by using conjugated nanoparticle, FSH33-
NP, from a peptide derived from FSH (Zhang et al., 2009)
This novel delivery system is shown to have very high selectivity
& efficacy for FSHR-expressing tumor tissues

32. An emerging technology that allows a fresh examination of the way
assisted reproduction is performed
Micro/nanofluidic devices potentially allow the development of more in
vivo like systems for the cell culture, and embryo development
Microchannel culture systems provide an embryo culture environment
that more closely mimics the in vivo environment & help in faster
embryonic development
Loading wells for embryo manipulation
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33.  In vivo, the embryo is bathed in a constantly changing
environment as it moves through the oviduct to the uterus
Microfluidics is well-suited to meet the needs of embryo culture
Changing of media is straightforward & no manipulation of embryo
is necessary
The media can be gradually changed around the embryo, rather
than subjecting it to sudden changes in environment
“lab-on-a-chip”

34. Zona pellucida removal device
(A) Microfluidic device used for insemination
(B) Microchannel that holds the oocyte at a specific point while allowing
media containing sperm to flow to & past the oocyte
(C) oocytes fertilized with microfluidic insemination that have developed
to cleavage-stage embryos
Microfluidic device designed for
microinsemination

35. Studies designed to separate motile sperm using microfluidic
devices have been reported
(Seo et al., 2007)
Development of efficient, reliable culture systems for single
embryos or gametes will provide new avenues to study the
microenvironment experienc-ed by these unique cells

37. “A chemical sensing device in which biologically derived
recognition entity is coupled to a transducer, to allow
quantitative development of a complex biochemical parameter”
“An analytical device incorporating deliberate & intimate
combination of a specific biological element (that creates a
recognition event) and a physical element (that transduces the
recognition event)”
Malhotra, (2005)
Definition of biosensor…

38. “Biosensor” signifies that the device is a
combination of two parts: (i) a bio-element, (ii) a
sensor-element
Biosensors can be of many types such as:
(i) Resonant biosensors,
(ii) Optical-Detection biosensors,
(iii) Thermal-Detection biosensors,
(iv) Ion-Sensitive biosensors, &
(v) Electrochemical biosensors

39. Specific “bio” element recognizes a specific analyte & the
“sensor” element transduces the change in the biomolecule
into an electrical signal
Responds to the presence of a specific analyte by
producing an electric signal that is proportional to the
concentration of analyte
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40. Quantitative analysis of progesterone in milk
 A biosensor for on-line measurement of progesterone in milk
to precisely detect estrus
 Novel P4 biosensor system which can sample milk from the
line on demand by the computer & pulled it to a central point
where it can be analysed with a disposable biosensor
(Frost, 1997)
 Another biosensor based on immunoassay for molecular
recognition, which was developed to run in approximately 8
minutes Claycomb et al.,
(1998)

41. Sensors fabricated by depositing anti-progesterone monoclonal
antibody onto screen-printed carbon electrodes (SPCEs)
Device operates in a competitive immunoassay format & relies on a
reduction in the binding of ALP -labelled P4 to sensor surface in
presence of endogenous milk progesterone (Pemberton et al.,
2001)
The results from field-tests show that the progesterone biosensor
can characterize the ovulation cycles of cows and detect pregnancy
Scientists are also trying to develop a fully automated ovulation
prediction system for dairy cows

42. Novel Biosensor for Rapid Measurement of
Estrogen Based on a Ligand-Receptor Interaction
A bioaffinity sensor for detection of estrogen, based on
specific binding of estrogen to its receptor immobilized on a
gold disk electrode
Sensitivity:- 10-50 pg

43. Reproductive management is a major financial concern of the
dairy industry, with missed estrus detection a main cause of lost
income
Use of nanotubes for breeding management is an interesting area
and can be exploited for prompt heat detection (Scott, 2005)
Nanotubes have the ability to detect analytes/hormones
could be used for assessing hormonal state of farm animals in
vivo to better control the timing of breeding
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44. Nanotubes implanted under the skin provide
real-time measurement of estradiol level in blood
These tubes have the ability to bind & detect the estradiol
levels at the time of oestrus by near infrared fluorescence
The signal from sensor could be incorporated as part of a
central monitoring
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45. DETECTION OF ANTISPERM ANTIBODY
By electrochemical method based on Au nanoparticles with a
mixed monolayer for eliminating nonspecific binding
Impedance spectroscopy is used to characterize the modified
procedures, the immobilization of sperm antigen (SpAg) & binding of
AsAb
The change of electron-transfer resistance correlates with
concentration of AsAb with the detection limit of 10 mU/ml

46.  Application of miniaturization technologies to medicine &
biology is progressing rapidly
 Nanotechnology will leave no field untouched by its ground
breaking scientific innovations
 These systems have the potential to perform high
throughput analysis, utilize small amounts of samples &
efficient analysis of gametes or embryos

47.  So far, the use of nanotechnology in animal sector has been
mostly theoretical, but it has begun and will continue to have a
significant effect in the main areas of the medicine &
reproduction
 Nanotechnology tools like microfluidics, nanomaterials,
bioanalytical nanosensors have the potential to solve many more
puzzles related to animal health, production & reproduction
 significant benefits could arise from nanotechnology applied to
animal reproduction; but.... there is a need to prepare for
oversight, health & environmental safety, societal issues that are
likely to arise from these applications