Chick Embryology

INTRODUCTION
Embryology is the study of the development of organisms. This is as true of plants as it is of

animals. In all cases a cell or a group of cells becomes separate from the organism as a whole,
either physically or physiologically, & progressively becomes a new complete organism. For
example- a microscopic cell, the human egg, proliferate & develops into a new highly
complex multicellular organism. This is known as development .It involves three
fundamental processes:a. Increase in cell number by cell division(cleavage), b.The
differentiation of cells into tissue(cell differentiation & tissue formation),c.The
arrangement of these cells & tissues to form parts & organs(organogeny).The study of the
developmental process from a single cell to a baby in 9 months. In other words,
investigations of the molecular, cellular and structural factors contributing to the formation of
an organism.Developmental biology is another name of embryology.The literal meaning of
embryology is the study of embryo where embryo denotes the juvenile stage,either within the
shell,egg membrane or in the mother’s womb.It deals with the different stages of an
organism.Developmental biology, on other hand,is not only restricted to different
developmental stages of an embryo but also includes the biochemical,genetic& physiological
& morphological aspects of the developing embryo.
HISTORICAL REVIEW.
Aristotle (384-322 B.C), a Greek philosopher studied embroys of animals and many of his
observations were accurate. Anaximander (600 B.C) was perhaps the first to conceive the
idea that living creatures arose from moist element and man was like fish in the
beginning.The Greek philosopher Empedocles was aware of the fact that foetus arose partly
from male and party from females semen. He also had knowledge regarding the different
stages of development and that heart formed first and nails later. The credit of establishing
Embryology as a separate and independent branch of Biology goes to Aristotle.He believed
that the soul or form of the embryo was derived from the father and mother supplies the
matter or soil in which the embryo grows. Aristotle’s views were acceptable through most of
the medieval period, and his views appeared in the writings of Fabricius (1537-1619) and
Harvey (1578-1657).
1 | Development of chick embryo as a model system for embryological study of Vertebrate

These scientists stated that the soul or the determining cause for development existed in the
blood of animals. With the invention of the microscope by RobertHooke in 1665 the sperm
and ovum were discovered but their importance in developmentwas not established.
One of the most amazing conclusions of modern biological research is that the mechanisms
of development are very similar for all animals, including humans. This fact has only been
known since it has become possible to examine the molecular basis of developmental
processes. As recently as 1980 we knew nothing of these mechanisms, but 25 years later we
know a lot. Over this period, developmental biology has been one of the most exciting areas
of biological research. These dramatic advances came from three main traditions that became
fused together into a single world-view: experimental embryology, developmental genetics,
and molecular biology. Experimental embryology had been in existence since the beginning
of the twentieth century, consisting mainly of microsurgical experiments on embryos of frogs
and sea urchins. These had demonstrated the existence of embryonic induction: chemical
signals that controlled the pathways of development of regions of cells within the embryo.
The experiments showed where and when these signals operated, but they could not identify
the signals, nor the molecular nature of the responses to them. Developmental genetics has
also existed for a long time, but it really flowered in the late 1970s when mass genetic screens
were carried out on the fruit fly Drosophila, in which thousands of mutations affecting
development were examined. These mutagenesis screens resulted in the identification of a
high proportion of the genes that control development, not just in Drosophila, but in all
animals. Molecular biology had started with the discovery of the three dimensional structure
of DNA in 1953, and became a practical science of gene manipulation in the 1970s. The key
technical innovations were methods for molecular cloning to enable single genes to be
amplified to a chemically useful quantity, methods for nucleic acid hybridization to enable
the identification of DNA or RNA samples, and methods for DNA sequencing to determine
the primary structures of genes and their protein products. Once this toolkit had been
assembled it could be applied to a whole range of biological problems, including those of
development. It was used initially to clone the developmental genes of Drosophila. This
turned out to be of enormous importance because most of the key Drosophila genes were
found to exist also in other animals, and frequently to be controlling similar developmental
processes. Molecular biological methods were also applied directly to vertebrate embryos and
used to identify the previously mysterious inducing factors and the genes regulated by them.

Out of over 1 million animal species, modern developmental biology has focused on a very
small number which are often described as “model organisms.” This is because the
motivation for their study is not simply to understand how that particular animal develops,
but to use it as an example of how all animals develop. Much developmental biology research
is supported by medical research funding bodies and their ultimate goal is to understand how
the human body develops, even if this is not the immediate goal of the investigators
themselves. For this reason there is often an attraction to studying a process that also occurs
in humans. A model organism is a species that has been widely studied, usually because it is
easy to maintain and breed in a laboratory setting and has particular experimental advantages.
 Model organisms are nonhuman species that are used in the laboratory to help
scientists understand biological processes.
 They are usually organisms that are easy to maintain and breed in a laboratory setting.
 For example, they may have particularly robust that are easily studied and
manipulated in the lab, this is useful for scientists studying development.
 Or they may occupy a pivotal position in the evolutionary tree, this is useful for
scientists
There are many model organisms. One of the first model systems for molecular biology was
the bacterium Escherichia coli, a common constituent of the human digestive system. Several
of the bacterial viruses (bacteriophage) that infect E. coli also have been very useful for the
study of gene structure and gene regulation (e.g. phages Lambda and T4). However,
bacteriophages are not organisms because they lack metabolism and depend on functions of
the host cells for propagation The six model organisms and their positions in the phylogenetic
tree of animals are shown in Fig. 1.1 It will be noted that they do not provide a very good
coverage of the animal kingdom, as there are just two invertebrates and four vertebrates.
However, theevolutionary distance between them is sufficiently large to make it likely that
any developmental features possessed by all six are shared by the entire animal kingdom.
:T
able:2.1-Table showing species ,their common name including their phylum,subphylum and class

The six model species have each been selected because they have some particular
experimental advantages for developmental biology research. It is true that sometimes
individual scientists choose their organisms just because they like looking at them and
working with them. But there are also some more objective practical considerations that
govern the selection of organisms for particular purposes which are summarized in figure.
Fig: 1.1-Phylogenetic tree showing the positions of the big six model organisms used in developmental biology
The Chick as Laboratory Material- The chick is one of the most , satisfactory
animals on which student laboratory work in embryology may be based. Chick embryos in a
proper state of preservation and of, the stages desired can readily be secured and prepared for
study. Used as the only laboratory material in a brief course they afford a basis for
understanding the early differentiation of the organ systems and the fundamental processes of
.body formation common to all groups of vertebrates. In more extended courses where
several forms are taken up, the chick serves at once as a type for the development
characteristic of the large-yolked eggs of birds and reptiles, and as an intermediate form
bridging the gap between the simpler processes of development in fishes and amphibia on the
one hand and the more complex processes in mammals on the other. In medical school
courses where a knowledge of human embryology is the end in view the chick not only
makes a good stepping-stone to the understanding of mammalian embryology, but also
provides material for the study of early developmental processes not readily demonstrable in

mammals.The fowl belongs to order Galliformes of class Aves of subphylum Vetebrata. Its
embryology has been worked out more extensively because of the following reasons:
 Its eggs are of convenient size.
 Its egg are easy to procure.
 Its earlier stages are similar to those of reptiles, mammals and even of man.
 Its different developmental stages can be easily obtained in laboratory conditions for
experimental purposes.
Despite of above reasons, the embryology of chick is important due to its phylogenetic
significance. The eggs and embryos of chick have to endure dryness and the harshness of
rugged surroundings .Therefore, they are supplied with a watery envelope of albumen, tough
outer membranes, and often a hard shell. The newly hatched young should be well developed
to crawl on land and defend themselves. This requires a large store of nutrient in the egg in
the form of yolk. Further more, the embryos need special adaptations to protect their delicate
outer surfaces; to carry on respiratory exchange of gases with the surrounding air and needed
receptacles to retain the wastes which , if accumulate with in the embryo proper become
toxic. To serve these functions , the embryo developed four foetal membranes namely , the
chorion , amnion , yolk sac and allantois. Eggs which meet these specifications are known as
cleidoic eggs characteristic of reptiles , birds and monotremes.
EMBRYOLOGY OF CHICK
The chicken (taxon -Gallus gallus) embryo develops and hatches in 20 to 21 days and has
been extensively used in embryology studies. Historically, the chicken embryo was one of the
first embryos studied, readily available and easy to incubate, embryo development can be
directly observed by cutting a small window in the egg shell. A key to this model organism
study was the establishment of a staging atlas by Hamburger & Hamilton in 1951, which
allowed specifc developmental landmarks to be seen and correlated with experimental
manipulations of development. This much cited paper included images of all key stages and
was more recently republished in the journal Developmental Dynamics (1993), for a new
generation of avian researchers. Probably just as important has been the recent chicken
genome sequencing, providing a resource to extend our knowledge of this excellent
developmental model.

Fertilized eggs can be easily maintained in humidified incubators and during early stages of
development the embryo floats on to of the egg yolk that it is using for nutrition. As the
embryo grows it sinks into, or below the, yolk. The regular appearance of somites allowed
early experimenters to acurately stage the embryo. The embryo was accessible and easy to
manipulate (limb grafts/removal etc) that were informative about developmental processes.
Chicken cells and tissues (neural ganglia/fragments) are also easy to grow in tissue culture.
The discovery that quail cells have a different nuclear appearance meant that transplanted
cells (chick/quail chimeras) could be tracked during development. For example, LeDourian's
studies showed how neural crest cells migrate widely throughout the embryo.
GONADS & GAMETES

Primary sex differentiation in the embryo includes differentiation of the initially indifferent
gonads into either testes in genetic males or ovaries in genetic females, and differentiation of
the primordial germ cells (PGC) into either spermatogonia or oogonia. The interaction
between germ cells and somatic cells is essential for the initiation of the latter process,
gametogenesis, in both birds and mammals.
The embryology starts with the production of gametes from gonads & union of male &
female gametes during fertilization. The gonads & gametes of all animal including chick are
different in case of male & female.
Testes & Sperm: These are seen in case of male chicks. In male fowl,the testes are paired
ovoid structures,suspended from the dorsal body wall by mesorchia. Sperm maturation occurs
inside the seminiferous tubules of testes. A vas deferens arising from each testes carries
sperms into the cloaca. The cloacal opening lies on a genital papilla. A mature sperm of fowl
is about 50μ in length. It’s head is short & cylindrical with a pointed acrosome. The tail is
long,tapering&vibratile.

Fig:1.2-Figure showing reproductive organs of male fowl
Ovary & Ovum: In all flying birds only one ovary of the left side is functional. The right
ovary & oviduct degenerate. It is advantageous for the birds because: Bird’s eggs are large
&megalecithal. The development of eggs in both the ovaries & their passage through both the
oviduct would adversely affect the size of the eggs & put a limitation on the amount of
reserve yolk. Absence of one ovary & oviduct makes the body lighter & more efficient for
flight.The female fowl(hen) has only one functional ovary & a single oviduct. It present on
the left side. The right ovary & oviduct are present in the young. In adult these are presented
by streak. Ovaries are suspended from the dorsal body wall by mesovarium. The primordial
germ cells differentiate in 5 days old chick embryo but sex cells are alike in the two sexes.
Fig 1.3-Figure showingreproductive organ of female hen

The ovary consists of bundles of ovarian follicle embedded in the storma. Each ovarian
follicle has a large central oocyte surrounded by a peripheral layer of amall follicle cells.
Oocyte grows in size by the accumulation of yolk. The yolk is drawn from the maternal blood
stream by the follicle cells &is deposited in the ovum. While the yolk forms the 1 st sphere in
the centre of the ovum. The subsequent layers of yellow &white yolk are deposited
alternately in a daily cycle for a specific period during maturation of ovum.The ovary is a
cluster of developing ova, and is located midway between the neck and the tail of the bird
and attached at the back. The ovary is fully formed when a pullet chick hatches but is very
small until the chick reaches sexual maturity. At hatch, a pullet chick has tens of thousands of
ova, or potential eggs that theoretically could be laid, although most never develop to the
point of ovulation. The maximum number of eggs a hen can lay is determined when it
hatches because no new ova form after the chick hatches. Each ovum starts as a single cell
surrounded by a vitelline membrane. As the ovum develops, additional yolk forms. The color
of the yolk comes from fat-soluble pigments, called xanthophylls, contained in the hen's diet.
Hens fed diets with yellow maize (field corn) or allowed to range on grass typically produce
eggs with dark yellow yolks. Hens fed diets with white maize, sorghum, millet, or wheat
typically produce eggs with pale yolks. The color of yolks can be improved (made darker) by
the addition of marigold petals to feed to provide the desired level of xanthophylls. The ovum
is enclosed in a sac that ruptures along the stigma, or suture line, during ovulation.The
nucleus in the young oocyte lies in it’scentre. But with the deposition of yolk, the egg nucleus
is displaced towards the animal pole. The cytoplasm forms a germinal disc or blastodisc
around the nucleus & lies at top of yolk mass. Later on, zona radiata is formed as a thin
membrane inside the layer of follicle cells. In the stroma of ovary, the young & small follicles
are present in the centre, & larger & older ones on the periphery. The fully grown follicles
bulge into the coelom. The formation of the ovum, the phenomena of fertihzation, and the
stages of development occurring prior to the laying of the egg have been more completely
worked out in the pigeon than in the hen. The observations which have been carried out on
the hen's egg indicate, as might be expected from the near relationship of the pigeon and the
hen, that the processes in the two forms are closely comparable. The following account which
is based chiefly on observations made on the pigeon's egg may, therefore, be taken to apply
equally well in all essentials to the hen's egg. The part of the egg commonly known as the
"yolk" is a single cell, the female sex cell or ovum. Its great size as compared with other cells
is due to the food material it contains. While the egg cell is still in
the ovary, material which is later used by the embryo as food is deposited in its cytoplasm.
This deposit which is known as deutoplasm consists of a viscid fluid in which are suspended
granules and globules of various sizes. As the deutoplasm increases in amount the nucleus
and the cytoplasm are forced toward the surface so that eventually the deutoplasm comes to
occupy nearly the entire cell. This abundance of deutoplasm accumulated in the ovum
furnishes a readily assimilable food supply, which makes possible the extreemly rapid
development of the chick embryo.A section of the hen's ovary passing through a nearly
mature ovum . The ovum and the tissues which surround it projecting from the ovary but
connected to it by a constricted stalk of ovarian tissue. The protuberance containing the ovum
is known as a follicle. The bulk of the ovum itself is made up of the yolk. Except in the
neighborhood of the nucleus the active cytoplasm is but a thin film enveloping the yolk.
About the nucleus a considerable mass of cytoplasm is aggregated. The region of the ovum
containing the nucleus and the bulk of the active cytoplasm is known as the animal pole
because this subsequently becomes the site of greatest protoplasmic activity. The region
opposite the animal pole is called the vegetative pole because while material for growth is
drawn from this region it remains itself relatively inactive.

Fig 1.4-Reproductive tract of hen showing the
ovulation process
Oviduct: When ovulation occurs, the ovum (yolk) enters the oviduct. The oviduct is a
twisted tube that is 25 to 27 inches long when fully developed and is divided into five major
sections. These sections are the infundibulum, magnum, isthmus, shell gland, and vagina.
The first part of the oviduct, the infundibulum (or funnel) is 3 to 4 inches long and engulfs
the ovum released from the ovary. The term funnel is an inaccurate name for this section
because it suggests that the infundibulum is waiting for the yolk to fall into it, which is not
the case. Instead, the released yolk stays in place, and the muscular infundibulum moves to
surround it. The yolk remains in the infundibulum for 15 to 17 minutes. Fertilization, if it is
going to occur, takes place in the infundibulum.The next section of the oviduct is the
magnum. At 13 inches long, it is the largest section of the oviduct, as its name implies
(magnum being the Latin word for "large"). The yolk remains here 3 hours, during which
time the thick albumen (egg white) forms.The third section of the oviduct is the isthmus,
which is 4 inches long. The isthmus, as its name implies, is slightly constricted (the term
isthmus referring to a narrow strip of land joining two larger tracts of land). The isthmus is
where the inner and outer shell membranes form. The developing egg remains here for 75
minutes.
The next section of the oviduct is the shell gland (or uterus), which is 4 to 5 inches long. In
this section, the shell forms on the egg. The shell largely is made of calcium carbonate. The
hen's body mobilizes 8 to 10 percent of body calcium from its bones to make the egg's shell.
Bone calcium provides 47 percent of the calcium required to make a shell, and the hen's diet
provides the remainder. Pigment deposition, if there is any, occurs in the shell gland. The egg
remains here for 20 or more hours.The last part of the oviduct is the vagina, which is about 4

to 5 inches long. The vagina does not really play a part in egg formation but is important in
the laying of the egg. The vagina is made of muscle that helps push the egg out of the hen's
body. The bloom, or cuticle, forms on the egg in the vagina prior to oviposition (the laying
of the fully formed egg).
Fig 1.5-Figure showing reproductive tract of laying hen.

The egg travels through the oviduct small end first but turns in the vagina and comes out
large end first.Near the junction of the shell gland and the vagina are deep glands known as
sperm host glands that can store sperm for long periods of time, typically 10 days to 2 weeks.
(One of the unique things about birds is that the sperm remain viable at body temperature.)
When a hen lays an egg, sperm can be squeezed out of these glands into the oviduct and then
can migrate to the infundibulum to fertilize an ovum.
OVULATION & MATURATION:

The fully developed oocyte breaks through the ovarian wall, ruptures the follicle & comes out
into the coelom near infundibulum. During ovulation the muscle fibres surrounding the
mature follicle contract & egg bursts out of the capsule.
At this stage, the nucleus of the oocyte enters 1st maturation division. Secondary maturation
division occurs after ovulation, when secondary oocyte has reached the anterior end of
oviduct.
COPULATION & OVIPOSITION:
During copulation , the cock or rooster crouches on the back of the hen & while the latter
elevates her tail, & thus, turning the cloaca upward, the cock or rooster turns his tail
downward, so that the two cloaca becomes opposed to eachother. The flagellated
spermatozoa are then transferred & injected into the vagina of the hen. They travels upward

& accumulate in then upper portion of oviduct where they may remain alive for two weeks &
will fertilize the ova as the latter enter the oviduct.
FERTILIZATION:
Fertilization occurs in the anterior part of oviduct. After the complete development, the
ovarian follicle breaks through the ovarian wall,thus rupturing the follicular envelope &
liberating the immature ovum into the coelomic cavity near the ostium of the oviduct, without
the rupture of capillary vessels & the shedding of blood. Immediately prior to this, the 1 st
maturation division gives rise to a small sized 1 stpolocyte& a large sized secondary oocyte.
The secondary oocyte enters the oviduct by the active movements of ostium which virtually
grasps & swallows it.In the upper region of oviduct, many sperms surround the secondary
oocyte, the several of which enter the oocyte(polyspermy). Though polyspermy occurs as a
rule in chick, but the spermatozoon which succeeds 1st in penetrating it’s head through the
eggmembrane,brings effective fertilization. It’s nucleus fuses with the egg nucleus
( femalepronucleus )& thus amphimixis occurs. The nuclei of other sperms become
degenerated subsequently.The mere entry of the sperm in the egg membrane induces the
secondary oocyte to undergo quick second maturation division & become mature ovum. The
passage of the fertilized egg in the oviduct is slow, taking approximately 22 hours. As the
fertilized egg passes downward inside the oviduct, two event occurs simultaneously;firstly,
most of the early development of fertilized egg i.e., cleavage,blastulation& even gastrulation
takes place; & secondly, various accessory coverings are added to the developing egg by the
oviducal wall, before it is laid. The development of the chicken embryo occurs in the egg,
partly in the body of the hen but mainly during the brood period after the lay.The proteins and
lipids are synthesized in the liver of the hen before ovulation and are stored in layers in the
egg yolk. The mature egg (= oocyte) is released by the ovary during ovulation and is then
incorporated in the funnel-shaped opening (infundibulum) of the oviduct. Fertilization by
the sperm of the cock can occur at that location during a short time period (approximately in
the first 15 minutes). Therefore, the sperm cells have to pass the whole way through the
oviduct from the cloaca up to the infundibulum. The sperm cells can be stored temporarily in
the vagina where selection is taking place. The egg membrane becomes the fertilization
membrane in a very short time after penetration by the sperm cell. This membrane inhibits
penetration by other sperm cells. The fertilized oocyte (= zygote) starts immediately with the
cleaving process and a small germ layer is formed at the surface of the yolk. The egg is

loaded with albumen at the magnum. The two semi permeable shell membranes are formed
at the isthmus while the poriferous calcified shell is set off in the uterus. The egg is layed
around 24 to 28 hours after fertilization. A light contraction of the egg mass takes place after
cooling down and results in the formation of an air chamber between the two aforementioned
shell membranes.
FORMATION OF ACCESSORY LAYERS AROUND THE FERTILIZED

OVUM.
As the fertilized egg passes down the first or convoluted & glandular part of
oviduct( magnum), a thick layer of dense albumen is secreted around the viteline membrane.
As the egg descends in the oviduct it rotates twisting the adherent albumen in spinal cords,
called chalazae, projecting at either ends of the yolk.
Additional albumen is deposited in concentric layers about the ovum while the egg moves
down in the oviduct. The shell membrane, which are sheets of matted organic(keratin)
fibres, are added to the egg as it progresses through the isthmus part of oviduct. The porous
calcareous shell is added while the egg passes through the shell gland portion of the oviduct
(the uterus). Within the egg, there forms a air space in between the two shell membranes at
the blunt end of the egg.According to Von Bear’s rule(1828), the axis of the embryo is
crosswise to the long axis of the egg. The sharp end of the egg of hen is on the embryos right,
blunt end is on it’s left. The chalazae becomes coiled clockwise on the future embryo’s left,
counter clockwise on it’s right. The chalazae help in keeping the ovum well placed in the
centre of the albumenous mass after it complete it’s formation.The oviduct leads ultimately to
the cloaca, a common chamber into which the excretory ducts also open. The developing egg
remains in the body approximately for 22 hours before it is laid. The development of eggs of
hen occurs at the body temperature, i.e., about 38°c.

Fig:1.6-Figure showing fertile and infertile eggs
STRUCTURE & CHEMISTRY OF THE FERTILIZED EGG :
The fully formed & newly laid fertilized egg of hen is a large epithelial body with one end
broader than the other. It is about 3 cm broad & 5 cm long with it’s enormous mass of yolk; it
is an example of polylecithal(macrolecithal) or highlytelolecithal egg. The polarity of an
egg is well marked, the animal pole having very small amount of active cytoplasm in the
form of disc & a zygote nucleus & is called germinal disc or blastodisc. Most of the space
of animal pole & vegetal pole of the egg is occupied by the yolk. The yolk &blastodisc are
ultimately bounded by a plasma membrane. Besides the plasma membrane, the egg is
bounded by no less than five accessory membranes. The plasma membrane is followed by a
vitelline membrane which is a converted form of zona radiate. According to balinsky(1970)
the vitelline membrane of hen’s egg is of double origin. The inner layer vitelline membrane is
produced in the ovary, in the space between the oocyte & the follicle cells.this layer is
composed of very rough fibres. An outer more finely fibrous layer is then formed on the top
of the 1st when the egg enters the upper portion of the fallopian tube. The next egg membrane
is the white or albumen of the egg. Yolk & egg whit provide food for the developing embryo.
Towards the end of the incubation, a part of the shell dissolves & the CaCO 3 released is
utilized to build the developing bones. The shell also provides a little protein. The egg whit or
albumen supplies protein,water& minerals, while enzymes, vitamins & pigments come both
from yolk & albumen.
Fig:1.7-Figure showing parts of hens egg

Egg shell:The egg shell is the outermost protecting covering. It is a whit, hard but porous
structure, formed of CaCO3. The pores have a diameter of 0.04 to 0.05 m & are about 7,000
in number. These are more numerous at the broad end of the shell & are filled with some
proteinaceous substance related to collagen. Thus the shell is porous & allows exchange of
oxygen & carbon dioxide through it. The shell is secreted by shell gland portion of the uterus.

The shell is soft & flexible in afreshly laid egg but soon becomes hard & brittle. The type of
shelled egg shut off from it’s surrounding, is termed cleidoic.
Shell membrane: Inside the shell are two shell membranes. These are formed of keratin
fibres matted together & are deposited by the isthmus part of oviduct.
Fig:1.8-Figure showing parts of shell membrane of egg

The outer shell membrane is thick & tough while the inner one is thin. The two membranes
are in close contact throughout the inner surface of the shell except at the broad end, where
they are separated by a wide air space. The air space is filled with air & is formed when the
egg is about to be laid down. It helps the developing embryo in breathing. In the fresh egg the
air space is very small. It increases in size with the age of egg due to evaporation of water
through porous shel.
Shell & membrane of hen’s egg.
Albumen: The space between the shell membranes & vitelline membrane of the ovum is
filled with albumen or the white of egg. It consists of 85% water & the rest 15% is a mixture
of several proteins, mostly albumins which make up 94% of the dry weight. The egg
albumen is separated into two parts. The outer part is watery & is known as light albumen
or less dense albumen. It is secreted partly by the isthmus of oviduct & partly by the
uterus. The middle layer of albumen is thick, gelataneus, semisolid & viscous & known as
dense albumen. It is secreted by the glandular wall of magnum part of oviduct. The
innermost layer is made of very viscous albumen(glycoprotein) called the chalaziferous
layer which surround the ovum proper. It forms a pair of spirally twisted ropes or cords
called the chalazae, one towards each pole of the egg. Chalazae helps to keep the ovum in
the centre of the egg albumen. The albumen plays a role in nutrition of the embryo, it serves
as a water store& it also provides ednvelope protecting the embryo from mechanical &
chemical injuries.

Ovum: The ovum is a large spherical body embedded in the egg albumen. It is surrounded
by a plasma membrane & a vitelinemembrane .the vitelline membrane is a modified zona
radiate & has a double origin. It’s inner layer is formed in the ovary & is composed of very
rough fibres, while the outer layer is formed in the upper portion of the fallopian tube & is
formed of fine fibres.
Yolk: The major portion of ovum is formed of yolk. The ovum contains a nucleus surrounded
by a very small amount of yolk free active cytoplasm restricted to a tiny germinal disc or
blasdtodisc,whichfloates at the animal pole of the ovum. It is about 3 mm in diameter. The
blastodisc or germinal disc represents the living portion of the ovum & contains the zygote
nucleus which undergoes cleavage. It is whitish in colour& circular in outline. The somewhat
darker area around the white blastodisc is known as periblast or marginal area. The remaining
space of the animal & the entire vegetal hemisphere of the egg is thus occupied by the yolk.
The yolk is not homogenous but consists of alternate layers of yellow & white yolk arranged
concentrically around a central flask shapped mass of white yolk. The layers of yellow yolk
are thicker than those of the white yolk. The yellow colour of yolk is due to the presense of
yellow carotenoids. Apart from the difference in colour, microscope examination reveales
that there are differences in the glandular & globules of the two types of yolk. Those in the
white yolk being smaller & less uniform in appearance. The central flask shapped mass of
white yolk is termed, latebra, while it’s outer neck like part is known as the neck of latebra,
which expands under the surface of blastodisc into the broad area, called the nucleus of
pander.Most of the yolk is liquid, but abot 23% of it is solid called yolk sphere. The yolk as a
whole contains 48.7% water, 32.6% phospholipid & fats, 16.6% proteins,1% carbohydrate &
1.1 % other chemical molecules including vitamin A, B,D & E. the proteins of this yolk
occurs in the form of a phosvitin&lipovitelline. The lipids are the greater part of the solid
constituents of the yolk & serve as the major source of food for the embryo. About 50% fatty
portion of yolk is natural fat, & the rest is phosphatids& cholesterol. The carbohydrates of the
yolk are free glucose & polysaccharides combined with the phospholipids, phosphoproteins
&cerebrosides.

Fig:1.9Longitudinal section of hen’s egg.
CLEAVAGE OR SEGMENTATION.
MEANING OF CLEAVAGE:
Fertilization results into the formation of zygote. The process of segmentation (cleavage)
immediately follows fertilization or any other process which activates the egg. Cleavage
consists of division of the zygote into a large number of cellular entities. The cells which are
produced during segmentation are called blastomeres. At first, the cells remain closely asso-
ciated, but later on they form the lining of a hollow sphere called blastula. The blastula
contains a cavity named blastocoel and its outer covering is designated as blastoderm. The
formation of blastula culminates the cleavage period. The process of segmentation prepares
the groundwork for the future design of the embryo by producing adequate number of cells.
The cleavage also establishes the fundamental conditions for the initiation of next
developmental stage —gastrulation.
PLANES OF CLEAVAGE:
During early cleavage, distinct geometrical relationships exist between the blastomeres, i.e.,
each plane of cell-division bears a definite relationship with each other. The planes of
division are:
a. Meridional plane of cleavage: When a furrow bisect both the poles of the egg passing
through the median axis or centre of egg it is called meridional plane of cleavage. The
median axis runs between the centre of animal pole and vegetal pole.
b. Vertical plane of cleavage: When a furrow passes in any direction (does not pass through
the median axis) from the animal pole towards the opposite pole.
c. Equatorial plane of cleavage: This type of cleavage plane divides the egg halfway
between the animal and vegetal poles and the line of division runs at right angle to the
median axis.
d. Latitudinal plane of cleavage: This is almost similar to the equatorial plane of cleavage,
but the furrow runs through the cytoplasm on either side of the equatorial plane.
Types of Cleavage:
Considerable amount of reorganisation occurs during the period of cleavage and the types of
cleavage depend largely upon the cytoplasmic contents.
Different types of cleavage encountered in different eggs are catalogued below:
a. Holoblastic or total cleavage: When the cleavage furrows divide the entire egg. It may
be:
Equal: When the cleavage furrow cuts the egg into two equal cells. It may be radially
symmetrical, bilaterally, symmetrical, spirally symmetrical or irregular.Unequal: When the
resultant blastomeres become unequal ir size.
b. Meroblastic cleavage: When segmentation takes place only in a small portion of the egg
resulting in the formation of blastoderm, it is called meroblastic cleavage. Usually the
blastoderm is present in the animal pole and the vegetal pole becomes laden with yolk which
remains in antihcleaved state, i.e., the plane of division does not reach the periphery of
blastoderm or blastodisc.
c. Transitional cleavage: In many eggs, the cleavage is atypical which is neither typically
holoblastic nor meroblastic, but assumes a transitional stage between the two.
EFFECT OF YOLK ON CLEAVAGE: The fertilized egg in most cases contains
yolk, which are inert bodies. During division these bodies exert mechanical influences. In the
egg of Amphioxus, the yolk is thin and remains uniformly distributed. Therefore the division
is complete and early divisions occur at a very quicker rate. The amphibian egg contains yolk
which is localised at the vegetal pole. Here division initiates from the animal pole and
extends towards the vegetal pole, where the progress of cleavage slows down considerably.
Consequently, the animal pole divides faster than the vegetal pole. The eggs of reptiles and
birds are fully laden with large masses of yolk, thus restricting the cytoplasm and nucleus on
the periphery as a circular disc on the animal pole. Here the lines of cleavage divide only the
small animal pole region. Such effects of yolk on cleavage pattern influence the pattern of
further development.
CLEAVAGE & BLASTULATION ON CHICK:
Cleavage in hen’s egg begins while it passes down the oviduct even before it is surrounded
with albumen & other membrane. Because of much larger amount of inert yolk, the cleavage
furrows never cut through the enter ovum & divide the egg completely, therefore, the

cleavage divisions remain restricted to the small whitish cytoplasmic disc (blastodisc) which
occupies a small area at the top of the yolk at the animal pole. Such a type of claeavage is
known as partial or meroblastic & discoidal. The cell formed by meroblastc cleavage are
not completely demarcated but represent protoplasmic areas with separate nuclei, being
isolated by cleavage furrows. Moreover, the cleavage furrows do not extend the whole area of
blastoderm but are restricted to the center of blastodisc. This results in the demarcation of
cenral cells surrounded on the periphery by a rim of unsegmented cytoplasm. This
unsegmented area is known as periblast or marginal zone.
Fig:1.10Early cleavage in hen’s egg in surface view. A. First cleavage,B. second cleavage,C.Third cleavage, D.
Fourth cleavage,E. Fifth cleavage,F. Early morula.
First cleavage: The 1st cleavage furrow occurs at about 4½ to 5 hours after fertilization. It is
slightly meridional or vertical incison near the centre of germinal disc in the direction of the
animal-vegetal axis. It neither extends right across the disc, nor through it’s whole thickness,
so that the germinal disc is divided only incompletely & superficial into two blastomeres
which remain unseparated from the underlying yolk.
Second cleavage: In each of two blastomeres, mitotic spindles initiating the second cleavage
arise at right angle to the position which was occupied by the first cleavage spindle. This is
determines that the two simultaneously appearing second cleavage furrows will be at right
angles to the 1st . Since these two second cleavage furrows lie in the same plane & are
apparently continuous, they are usually considered together. This cleavage is also meridional.
Third cleavage: The cleavage furrows of third cleavage are vertical cutting across the second
set of cleavage furrows. These furrosws tend to be parallel to the 1 st cleavage furrows but are
usually irregular. Third cleavage results in the formation of eight blastomeres.

Fourth cleavage: It’s cleavage furrows are also vertical but not synchronous. The fourth
cleavage takes place in such a manner that the central part of the eight cells established by the
third cleavage are cut off from their peripheral portions. Thus, it proceeds to form eight
central blastomeres which are surrounded by eight marginal blastomeres. The central cells
now appear completely separated from one another in a surface view of the blastoderm, but
sections show them still unseparated from the underlying yolk at their lower ends & the
blastomeres on the circumference are, in addition , continuous with the uncleaved cytoplasm
at their outer edges.
CENTRAL & MARGINAL CELLS:
After fourth cleavage, the planes of further cleavage furrows become irregular, rapid &
involve both central & result in the formation of following cells:
(i).The apical or central cells that are located centrally in the blastodisc. These cells have
complete cell bounderies but are still continuous with the underlying yolk.
(ii).The radial furrows that further divide the marginal cells that lie on the periphery of the
central cell. These cells have incomplete cell boundaries being connected with the marginal
periblast on the sides & with the yolk on the undersurface.
MORULA & PERIBLAST FORMATION:
After 32 cell stage , the horizontal or latitudinal cleavage furrows are laid down 1 st in the
region of central cells & later in the marginal zone. This establishes a superficial single layer
of cells completely separated from the egg cytoplasm beneath. This layer form the
blastoderm. It’s cells rest upon a layer of cells which are continuous on their deep faces with
the underlying yolk. The lower layer of incomplete cells is called centralperiblast. The
marginal cells, which are present along the maegins of blastodisc remain continuous with
each other, are also continuous with the underlying yolk. These form marginal periblast.
In addition to the cleavages which are indicated on the surface, vertical sections of the 32 cell
embryo show cleavage planes of an entirely different character. These cleavages appear
below the surface & parallel to it. They establish a superficial layer of cells which are
completely delimited & rest upon a layer of cells which are continuous on their deep faces
with the yolk. Continued divisions of the same type eventually establish several strata of
superficial cells. This process appears 1st in the central portion of the blastoderm, & then
progresses centrifugally as the blastoderm increases in size but does not extend to it’s
extreme margin. The peripheral margin of the blastoderm remains a single cell in thickness &
the cell there lie unseparated from the underlying yolk as well continuous with the uncleaved

cytoplasm at thir outer edges. As the nuclei of the cells lying at the edges( marginal cells)
divide, more & more cells are cut off to join the ones lying in the centre.
Fig.1.11Blastodisc of hen’s egg in blastula showing subgerminal cavity, area opaca,area pellucid.
At the end of segmentation of chick embryo has arrived at the morula stage which consists
of the disc shapped mass of cells several stara in thickness lying closely applied to the yolk.
In the centre of the blastoderm the cells are smaller & completely defined; at the periphery
the cells are flattened,larger in surface extent, & are not walled off from the yolk beneath &
closely adherent to it,is called the zone of junction.
BLASTULATION:
In the latter stage of cleavage, blastomeres of the central area of blastodisc become separated
from the underlying yolk as under-
The cell division at this stage occurs in horizontal or tangential planes so that the upper cells
of the central area of the blastoderm become completely separated from the lower
incompletely separated cells which retain their connection with the yolk mass. The marginal
cells, which remain continuous with eachother around their outer edges, are also continuous
with the mass of yolk as well as with lower layer of cell resulting from horizontal division of
central cells. All these blastomeres eventually loose their dividing furrows & form
asyncytical layer of cytoplasm containing many nuclei which remain for a time around &
beneath the perimeter of the blastoderm. Eventually,a slit like cavity called the subgerminal
space or segmentation cavity, appears in between a thin discoidal cap of cells & the
underlying yolk.
The blastomeres of central area are further divided by many vertical & horizontal cleavage

furrows, so that the blastodisc has
four to five cells thick central
region. It forms a delicate layer of
rounded & somewhat loose cells
& is little over 3 mm in diameter.
The marginal cells or the cells of
the periphery (marginal periplast)
& the syncytical layer lying
underneath the sub germinal
cavity (central periplast) remain
closely adhere to the yolk & is
called periplast, which acts as
trophoblast tissue. It is supposed
to breakdown the yolk & making
it available to the growing
embryo.The peripheral area of
Fig:1.12Blastulation
blastoderm is opaque because the large yolk laden blastomeres rest directly on the underlying
yolk. This outer zone is known as the area opaca. The cells of the central mass of blastomeres
which overlie the fluid filled sub germinal cavity, on the other hand , are smaller & relatively
free from yolk, apper translucent. They constitute the area pellucida containing formative or
embryonic cells which later form the embryo proper, & the area opaca develops into extra
embryonic structures. pellucid segregate & move inward in the subgerminal space one by one
or in loose clusters & form a less organized thin flat epithelial layer called hypoblast. The
yolk poor & small sized blastomeres remain at the surface & organize into a thin superficial
layer known as the epiblast. The hypoblast remains separated from the epiblast by a narrow
slit, the blastcoel. The
process of separation of the hypoblast forming blastomeres is an act of infiltration,
polyinvagination,delamination or involution. In hen the hypoblast starts forming just before
the egg is laid, & is completed in the first hours after laying.
The two layers epiblast &hypoblast,certainly do not represent ectoderm & endoderm
respectively. In fact, the epiblast or the upper layer gives rise to all three germinal layers of
the embryo( the ectoderm, mesoderm & endoderm) & the hypoblast or the lower layer to the

endoderm. It has,therefore, been concluded that the two layered embryo of chick is still in the
blastula stage & that the lower layer, the hypoblast, corresponds to the blastocoels of
amphibians & fishes. Thus,cleavage converts the germinal disc into a disc shapped blastula
(with the blastocoels in the central area alone) called the discoblastula which floats at the top
of yolk mass. It is at this stage the egg is laid.
DISCOBLASTULA:
In chick the fully formed or mature blastula is adiscoblastula. It consists of:
A central multicellular & multilayered blastoderm free from the underlying yolk & central
periblast. It forms area pellucid. Underlying blastocoels or subgerminal cavity thet separates
blastoderm from the underlying centralperiblast& yolk. Marginal periblast tissue present on
the periphery of central blastoderm. It is called germ wall. It may be differentiated into inner
zone of distinct cells which divide rapidly, contribute cells to the peripheral zone of growing
cellular blastoderm.
Area pellucid & area opaca: The peripheral area of discoblastula is opaque because the
large yolk-laden blastomeres rest direcly on the underlying yolk. This outer peripheral zone is
known as the area opaca. The cells of the central mass of blastomeres, overlying the fluid
subgerminal cavity, are smaller & relatively free from yolk & appear translucent. They
constitute the area pellucid.
Fate map of discoblastula: The blastoderm in discoblastula of chick consists of two parts
i.e. the area pellucid & area opaca. The area opaca (area vitelline) does not form any part of
the embryo proper. It forms non or extra embryonic ectoderm which during later
developmental stages forms extra embryonic membranes & blood vessels ( areavasculosa).
The area vasculoss is involved in the digestion of the yolk. Hence, the fate map is restricted
to area pellucid. Moreover , fate maps of both epiblast & hypoblast are to be made out
separately.The epiblast has material for ectoderm, mesoderm & also for endoderm. The
hypoblast is formed exclusively of endoderm material. The area pellucid is differentiated into
the following areas i.e. the anterior 2/3rd of area pellucid is prospective ectoderm consisting of
prospective epidermis & extra embryonic ectoderm, a crescentic area is prospective neurula
area., next to neural area is the prospective notochord., it is followed by prechordal
mesoderm., on the side of perichordal region is the lateral plate mesoderm.

Fig1.13Blastodisc showing separation of epiblast & hypoblast & formation of blastula by
delamination.
GASTRULATION.
Gastrulation is a phase in the embryonic development of animals where the blastula
reorganizes itself into a gastrula. It does this by folding itself inward . This is a critical point
in development because it is when the embryo transforms itself from a hollow sphere made
from a single layer of cells into a multi-layered structure. The layers are called the primary
germ layers; the endoderm, ectoderm, and mesoderm. Each species has its own uniqueness
when it comes to the process of gastrulation, but there are similarities that span the entire
animal kingdom. Frogs, chickens, and sea urchins are 3 species most studied by
developmental biologists and comparative embryologists.
Fig:1.14 the process of transformation of a single celled zygote into the gastrula.

Fig:1.15gastrulation changes the cell layer from one to three.
The Effect of Yolk on Gastrulation:
The process of gastrulation begins as soon as blastulation is accomplished. Gastrulation as it
occurs in birds is not difiicult to understand if one grasps its fundamental similarity to the
corresponding process in forms with scanty yolk. In Amphioxus, gastrulation is an
inpocketing of the blastula . A double layered cup is formed from a single layered hollow
sphere much as one might push in a hollow rubber ball with the thumb. The new cavity in the
double walled cup is termed the gastrocoele. The opening from the outside into the
gastrocoele is called the blastopore. In gastrulation the single cell layer of the blastula is
doubled upon itself to form two layers. The outer cell layer is known as the ectoderm and the
inner layer as the entoderm. These layers differ from each other in their positional
relationship to the embryo and to the surrounding environment. Each has different functional
potentialities and each will in the course of development give rise to quite different types of
structures and organs. It is the importance of their later history rather than any complexity or
veiled significance about the way in which they arise that attaches such importance in
embryology to the estabhshment of these two layers.In the gastrulation of Amphibian
embryos the yolk forces the invagination of the blastoderm toward the animal pole, but the
inpocketing takes place into the blastocoele and the interrelationships of ectoderm, entoderm,
and gastrocoele are established in fundamentally the same way as in Amphioxus.
Gastrulation in birds is greatly modified by the large amount of yolk present . Infolding must
be effected in a disc of cells resting like a cap on a large yolk sphere. The smallness of the
blastocoele sharply restricts the space into which the invagination can grow. Instead of
arising as a relatively large circular opening the blastopore appears as a crescentic slit at the
margin of the blastoderm. The crescentic blastopore may be regarded as a potentially circular
opening which has been flattened as it develops between the growing disc of cells and the
unyielding yolk which under lies them. The invaginated pocket of entoderm which grows in

from this compressed blastopore is also flattened, conforming to the restrictions of the shape
and size of the blastocoele.Moreover the floor of the invagination is represented only by a
few widely scattered cells lying upon the yolk. It is as if the lower layer in its ingrowth was
impeded and broken up by the yolk. The scattered cells representing the floor of the
invagination soon disappear and the yolk itself comes to constitute the floor of the
gastrocoele. Notwithstanding the great displacement of the blastopore and the gastrular
invagination toward the animal pole and the restricted size and incomplete floor of the
gastrocoele, the cell layers and the cavity established can be homologized with the
corresponding features in forms where the course of development has not been so extensively
modified by yolk. A comparative review of the diagrams will afford a general understanding
of the infolding process of gastrulation. These diagrams aim to convey merely the scheme of
the process. They are therefore simplified and emphasize the similarities of gastrulation in
forms with widely varying amounts of yolk, rather than the details of the process in any one
form. With this general groundwork we may now profitably return to the blastula stage and
consider in somewhat more detail the process of gastrulation as it occurs in birds. Hen lays
egg while the embryo is preparing for gastrulation. In chick, the process of gastrulation is
prolonged & highly modified due to the presence of yolk. The gastrular movements
responsible for the establishment of embryonic membranes are convergence, streaming,
involution, epiboly & elongation.One of the unique features of chick gastrulation is the
cellular rearrangement that occurs at the posterior end of the blastula and forms the primitive
streak, a thickening of the tissue. The cells making up Hensen’s node at the end of the
primitive streak elongate across the blastula. Later, the cells of Hensen’s node regress, paving
the way for the formation of the central nervous system. In the chick embryo, the cells of the
ectoderm go on to form the skin and neural tissue, endoderm cells line the respiratory and
gastrointestinal tracts, and the kidneys, circulatory system and skeleton are made from the
mesoderm cells.
Gastrulation includes the following two types of morphogenetic movements:
a. Emboly: It involves only the epiblast & includes convergence, invagination & involution
(ingression) through blotting process by the mesentoblast cells (i.e.. progenitors of both
endoderm & mesoderm). Thus, emboly is exhibited with the formation, elongation &
ultimate recession of the primitive streak & also the formation of the head process. It results
in the formation of embryonic endoderm, mesoderm & notochord.

b. Epiboly: It includes the overgrowth of ectoderm or epiblast and also of hypoblast( or extra
embryonic endoderm) over the yolk mass.
Gastrulation is separated into three distinct processes or phases which occur in a
chronological sequence. These are: Formation of endoderm, Formation of mesoderm &
Formation of embryonic axial structure or primitive streak.

OBJECTIVES
One of the greatest miracles of nature is the rapid transformation of a seemingly lifeless egg
into a new living organism. Egg hatching provides a rare opportunity to study the stages of
embryonic growth during the 21 days of incubation, and gives students a chance to relate the
stages of chick development to that of other embryos. Although chick embryos start to
develop as soon as they are formed in the hen’s body, many things can affect their speed of
growth and their ability to form healthy chicks.
A. To make direct and daily visual observations of living embryos;
B. To learn the anatomy and physiology of the egg and embryos;
C. To understand that chicken embryos are more readily available and less expensive to use
than frogs in the study of embryology, anatomy and physiology.
D. To gain a deeper interest in biology and science. This is a "hands on" project and is an
excellent opportunity for young people to appreciate the development of the embryo

REVIEW OF LITERATURE
When a hen’s egg is laid, development is already well underwayand a two-layered disc of
many thousands of cellslies on top of the yolk (Bellairs and Osmond, 2005). The seriesof
events that take place during the next day or so, whenthe egg is incubated, sets up the body
axes and defines where the various organs will form. The first visible sign that marks the
head to tail axis of the future embryo is the primitivestreak, a thin opaque band of cells that
extends from the edge of the embryonic disc. One of the first organ systems to develop is the
vascular system. Blood islands are seen soon after one day of incubation and the circulation is
established about a day later. All this time the embryo is growing and changing shape. The
major regions of the embryo become recognizable, e.g. head, trunk and tail, followed by
formation of specific organs such as limbs, eyes, lungs etc. during the third and fourth days of
incubation. During the final stages of development, from ten days until twenty or twenty-one
days when the chick hatches, there is considerable growth and also elaboration of
differentiated cells and tissues including ossification of the skeleton and formation of feathers
The sequence of chick development has been described as an illustrated series of
developmental stages by Eyal-Giladi and Kochav (1975) for the first seven hours of
allowing standardisation between researchers working on chick development. Furthermore
the detailed description of the developing anatomy of the chick embryo in ‘The Atlas of
Chick Development’ which builds on the Hamburger and Hamilton description of chick
development (Bellairs and Osmond, 2005) has allowed reliable reporting of manipulated
chick embryonic anatomy.
Classical experimental embryology

Major insights into developmental mechanisms have come through the ease of ablating
tissues and of making tissue grafts in early chick embryos. Thus one can cut out a small
piece of tissue from an embryo and then graft it to another site in the same embryo or to
another embryo. Such microsurgery can be carried out through a window made in the
shell, while the embryo is still lying in the egg. Experiments can also be performed on
early chick embryos placed in simple culture systems although, in these systems, the
embryos can only continue developing for short periods of time.
Identifying cell-cell interactions
One of the most challenging problems in embryonic development is to understand the

mechanisms that ensure that all the parts of the body form in their proper places.
Waddington’s classical experiment on chick embryos in the 1930s (reviewed in Stern and
Conrad, 2000) identified an organizer region – equivalent to the organizer that had recently
been discovered in amphibian embryos – which specifies the body plan. This region is the
knot-like structure at the tip of the primitive streak known as Hensen’s node which when
grafted ectopically in early chick embryos, induces the formation of a secondary body axis.
This discovery was of major significance not only in terms of understanding how the body
plan is laid down in the chick embryo but also because it illustrated that similar mechanisms
operate in different vertebrates. Indeed Waddington then went on to demonstrate that
mammalian embryos also had an organizer by grafting the node of a rabbit embryo
ectopically in an early chick embryo and showing that it too could induce a secondary axis.
Other crucial long range signalling centres that specify the pattern of structures that develop
in particular regions of vertebrate embryos were also first identified through grafting
experiments in chicks. These include notochord/ floor plate which specifies the pattern of
neurons that develop with respect to top to bottom (dorso-ventral) of the neural tube (van
Straaten et al., 1988, Yamada et al., 1991), the polarizing region of the limb bud which
specifies the number and pattern of digits (Saunders and Gasseling, 1968), and the isthmic
organizer, at the midbrain/hindbrain which specifies the pattern of the adjacent regions of the
brain (Marin and Puelles, 1994; reviewed in Wurst and Bally-Cuif, 2001). Experiments in
chick embryos have also demonstrated the existence of a myriad of very local cell-cell
interactions between adjacent tissues. An example of this type of local interaction occurs
between epithelial and mesenchymal tissues at many different times and places in the embryo
and epithelial-mesenchymal interactions are often both dynamic and bidirectional. The
discovery of such interactions stemmed again from the ease of locally ablating epithelial
tissues in chick embryos. It is also possible to cleanly separate epithelial and mesenchymal
tissues, then make recombinations, between, for example, the two tissues in different
orientations or tissues from embryos at different stages of development and graft the
recombined tissues back into the embryo. Thus, for example, in the developing chick limb
bud, ablation experiments revealed that interactions between the apical ectodermal ridge (the
epithelial thickening that rims the bud) and underlying mesenchyme are necessary for
outgrowth (Saunders, 1948; Summerbell, 1974), while limbs developing fromrecombinations
between a mesenchymal hull from a right wing bud and ectodermal jacket from a left wing
bud demonstrated that interactions between the ectoderm (epithelium) covering the sides of

the bud and the underlying mesenchyme control the development of the structures on the
upper and lower sides of the limb (MacCabe et al., 1974).
Tracing cell fate

Increasingly sophisticated cell labelling techniques have been used to trace cell fate and to
follow the movements of cells in chick embryos, again uncovering general principles
applicable to all vertebrate embryos. Chick/quail chimeras were first recognised in the late
1960s as powerful tools for tracing cell fate and a whole raft of discoveries has come from
their analysis (reviewed in Le Douarin, 2005). Such chimeras are made by grafting fragments
of tissue from quail embryos into chick embryos and then following the fate of the grafted
cells. Quail cells were traditionally recognised in chick embryos through differences in
staining properties, but, more recently, quail-specific antibodies have been used. Chick
embryos with such inter specific grafts are able to survive and even go on to hatch because
the operations are carried out before the immune system has developed. Chick/quail chimeras
have, for example, led to the elucidation of the fate of cells from the neural crest, including
the discovery that the crest gives rise to almost the entire peripheral nervous system. The
neural crest is a small population of cells that arises at the edges of the neural plate. When the
neural plate rolls up to form the neural tube, neural crest cells come to lie on top of the tube
beneath the ectoderm. By tracing the fate of quail cells within chick embryos, it became clear
that neural crest cells disperse along particular tracts within the embryo and give rise to a
wide range of tissues, not only to the dorsal root and enteric ganglia, but also to the pigment
cells of the skin, Schwann cells and, in the head, connective tissues. Most of the skull is
derived from neural crest (reviewed in Creuzet et al., 2005). More recently, mouse/chick
chimeras have been used to study the derivatives of the neural crest (Fontaine-Perus and
Cheraud, 2005) and the development of somites (reviewed in Fontaine- Perus, 2000). Somites
are segmented blocks of cellsrunning down the back of the embryo on either side of the
spinal cord and give rise to back muscle (and the muscle of the limbs and tongue) and
vertebrae. Another method of tracing the fate of cells in embryos is by locally applying
lipophilic dyes that label cell membranes.This method has been used in chick embryos to
trace short term movements of cells, the dye eventually becoming diluted by cell division.
Thus fate maps of embryos have been created during gastrulation, a process in which some
cells in the upper layer move to the primitive streak, then detach and move inside the embryo
to create the three main body layers (e.g. Hatada and Stern, 1994). In addition, the fate of

cells in older chick embryos in specific regions, such as the hindbrain (Fraser et al., 1990) or
the developing limb (Altabef et al., 1997; Vargesson et al., 1997) has been traced using
lipophilic dyes. Some of these experiments have revealed, rather surprisingly, the existence
of cell lineage restricted compartments similar to those previously found in insects. In the
chick hind brain, for example, the progeny of labelled cells are confined to just a single
rhombomere (one of the seven recognizable hindbrain segments seen in both chick and
mouse embryos) and do not straddle the boundaries with adjacent rhombomeres showing that
each rhombomere constitutes a separate compartment (Fraser et al., 1990). Even more
remarkably, the ectoderm in both limb and non-limb forming regions of the trunk of chick
embryos was found to be compartmentalised with respect to the back and front of the embryo
(Altabef et al., 1997). The compartment boundary running down the sides of the embryo
appears to be important for positioning the limbs. Evidence for a similar boundary has also
been discovered more recently in developing mouse limbs (Guo et al., 2003) showing that the
initial findings in chick embryos are more widely applicable. Labelling cells by expression of
Green Fluorescent Protein (GFP) which can be visualised in living embryos has also been
used to good effect in chick embryos . Gene constructs can be introduced into cells of chick
embryos by electroporation or by using retroviruses. In a recent set of experiments, primitive
streak cells electroporated with GFP were cut out of one embryo and grafted into host
embryos at different positions along the streak. Films of the movement trajectories of the
labelled cells showed that the behaviour of streak cells depended on the position in which
they were grafted, thus revealing the importance of local interactions in determining cell
migration (Yang et al., 2002).
Molecular basis of development

Retinoic acid and cell-cell signalling The first successful use of chick embryos to screen
formolecules that can modulate development resulted in thefinding that a vitamin A
derivative, retinoic acid, couldmimic signalling by the polarizing region in the limb bud.The
signalling properties of the polarizing region, a smallregion of mesenchyme cells at the
posterior margin of thelimb bud, were first uncovered by grafting experiments(Saunders and
Gasseling, 1968). When the polarizing regionwas cut out from the posterior margin of one
chickwing bud and transplanted to the anterior margin of a secondwing bud, an additional set
of digits formed from theanterior part of the wing bud that normally does not formdigits. This
formed the basis for an assay to screen substancesfor polarizing activity (reviewed in Tickle,

2002). Unexpectedly,it was found that small pieces of paper or beads soaked in retinoic acid
(Tickle et al., 1985; ) grafted to the anterior margin of chick wing buds could induce mirror-
image duplications like those obtained following polarizing region grafts. There is now good
evidence that endogenous retinoic acid plays a role in limb bud initiation and that, when
applied to the anterior margin of a chick limb bud, triggers a cascade of expression of genes
that encode cell-cell signalling molecules that control both digitnumber and pattern. It has
also emerged that retinoic acid plays important roles in many other regions of vertebrate
embryos including the central nervous system. Furthermore the use of small beads to apply
locally defined molecules, including proteins and inhibitors, to chick embryos has been
exceptionally powerful in probing development.
Genes and development

Up until about 15 years ago, very little was known about the molecules that mediate cell-cell
signalling and control cell fate in embryos. But since then there has been an explosion
in discovering developmentally important genes through Drosophila mutants, tumours and
clinical genetics. Although none of these genes was first identified inchickens, a new
repertoire of manipulations devised to modulate gene expression and function has meant that
chick embryos have been at the forefront in testing gene function.A landmark in
developmental biology was the discovery of the vertebrate Hox genes. These genes are
relatives of the Drosophila homeotic genes which, when mutated, lead to rather grotesque
phenotypes in which one part of the body is replaced by another. An example is the mutant
Antennapedia in which an antennna is replaced with a leg. In insects, homeotic genes are
clustered and arranged in sequence along the DNA (this cluster is split in two in Drosophila)
and successive genes from the 3 _ end of the cluster to the 5 _ end are expressed in
successive segments along the body of the insect from head to tail. The homeotic genes
encode proteins that contain a DNA-binding domain and regulate expression of other genes
and thus are responsible for determining the development of individual segments. In
vertebrates, including chickens, there are four clusters of related Hox genes that appear to
have undergone duplication during evolution. Furthermore as, in the fly, 3 _ genes appear to
be expressed at high levels towards the head end of the embryo while 5 _ genes are expressed
at higher levels towards the tail end ( Duboule and Dollé, 1989; Graham et al., 1989 for initial
analysis in mouse embryos).5 _ members of two of the vertebrate Hoxgene clusters are
expressed in the early limb bud of both mouse and chick embryos in overlapping domains

centred on the posterior tip of the limb where the polarizing region is located (Izipsúa-
Belmonte et al., 1991; ) and the first evidence that these Hox genes are involved in patterning
the limb came from experiments on chick embryos. Thus it was shown that manipulations
that lead to formation of extra digits from anterior cells, polarizing region grafts or beads
soaked in retinoic acid, also resulted in an ectopic pattern of Hox gene expression anteriorly
mirroring the pattern normally seen posteriorly. In addition, when one of the more 5 _
Hoxdgenes was inserted into a replication-competent avian-specific retrovirus which was
then used to infect chick limb buds so that the gene was expressed throughout the limb bud,
an additional digit formed and/or changes in digit morphology were induced (Morgan et al.,
1992). These results have recently been illuminated by genetic manipulations in mice
(Zakany et al., 2004). Many other developmentally important genes, including genes
encoding cell-cell signalling molecules, have been discovered through finding relatives of
Drosophila genes.One of these vertebrate gene families comprises the three hedgehog genes,
which are related to the Drosophila hedgehog gene and includes the Sonic hedgehog (Shh) .
Shhencodesa long range signalling molecule which turns out to be involved in the
development of virtually every part of a vertebrateembryo (reviewed in Ingham and
MacMahon, 2001). Chicken embryos have played a key role in dissecting the function of Shh
, beginning with the finding that Shh was expressed in several signalling centres in the chick
embryo, including the polarizing region of the limb (Riddle et al., 1993;) notochord and floor
plate (Roelink et al.,1994;Hensen’s node (Levin et al., 1995) and in the zona
limitansintrathalamica (ZLI), at forebrain/midbrainboundary (Kiecker and Lumsden, 2004).
Experiments in chick embryos showed that Shh, like retinoic acid, can act as a polarizing
signal in the developing limb. When cells expressing Shh or beads soaked in Shh were placed
at the anterior margin of a chick wing bud, mirror-image duplications of the digits developed
(Riddle et al., 1993; Yang et al., 1997). Furthermore, it was shown that beads soaked in
retinoic acid induced ectopic expression of Shh , thus providing a mechanism for the
induction of digit duplications by retinoic acid (Riddle et al., 1993). Shh was also shown to
mediate patterning of the neural tube by the notochord/floorplate and also to pattern the
adjacent somites. Shh is expressed asymmetrically in the chick node and was one of the first
genes to be shown to be involved in setting up Left/ Right asymmetry through a signalling
cascade includingthe secreted factors, Nodal and Lefty (Levin et al., 1995).

Genomics and chicken development
The recent breakthroughs in chicken genomic resources have reconfirmed the chicken as an
exceptional model organism for developmental biology. Sequencing of the chicken genome
has allowed forward genetics to be applied to a chicken developmental mutant for the first
time. We are now able to easily identify chicken homologues of genes known to act during
development in other species as well as to search for new developmentally regulated genes or
regulatory elements. The comprehensive EST databases will enable in silico expression
analysis and large scale microarray based expression profiling thus uncovering the
developmental regulation of gene expression on a genome wide scale. Transgenic
technologies in chicken also now allow manipulation of gene expression either transiently or
stably during development The embryo of the domestic fowl (Gallus gallus) holds the record
as the animal with the longest continuous history as an experimental model for studies in
developmental biology, spanning more than 2 Millenia. Throughout this time, it attracted
great naturalists, artists, philosophers, and pioneers of biology and stimulated them to think
about the most fundamental questions on generation and life like no other organism has ever
done, except the human. The ancient Egyptians are documented as having opened hens’ eggs
at different periods during incubation to observe the progress of embryo development, and by
around 300 BC Aristotle undertook careful studies of the morphology of the embryo (as
much as he could without the aid of magnifying devices); this can be considered as the first
‘scientific’ study of embryo development and his work referred to by hisfollowers right up to
the 19th Century (Needham, 1959). After the mediaeval ‘Dark Ages’, the resurgence of
aninterest in Anatomy and embryo development in the Renaissance attracted figures
including Leonardo DaVinci (1452–1519), UlisseAldrovandi (1522–1605) and Hieronymus
Fabricius ab Aquapendente (1537–1619) to return to the study of the embryo within the egg.
At this time the debate between Preformation and Epigenesis was starting to gather
momentum and the chick embryo was often a reference to which subscribers to either camp
resorted to support their favourite theory. As an attempt to contribute to the debate, William
Harvey (1578–1657) observed chick embryos at early stages of development and concluded
that the heart was the first functioning organ to develop in the embryo. By observing the
motion of the blood through the heart and early vessels, he discovered the circulation of the
blood and understood the function ofarteries and veins. The ensuing 200 years or so were
accompanied by an increase in the number and the detail of anatomical studies of the embryo,
each time enriched by a new technical advance. The introduction of histological sectioning

and of selective staining methods allowed Pander and von Baer to start to understand the
significance of germ layers in development. These pioneers also started to ask question about
causality in development—what mechanisms are responsible for such stereotyped
development? Further andincreasingly careful histological studies followed throughout the
19th Century, with the most important contributions being made by Rauber and Hensen and
culminating in a beautiful histological atlas by Mathias Duval (1889). By the end of the 19th
Century, experimental embryology (Entwicklungsmechanik) started to replace simple
histological observation as it became clear that principles could only emerge from
experimental manipulation of the embryo, but the initial advances were mainly made through
work in other organisms (sea urchins and amphibians for ‘embryoembryonic regulation’ and
induction, marine invertebrates for lineage studies, Drosophila for developmental genetics)
and the chick was a little slower in catching up. But therewere some salient chick studies at
this time, including Graeper’s spectacular 3-d stereo time-lapse movies ofembryos labelled
with spots of vital dyes to follow cell movements (made in 1926, published in 1929
andevolution from the wild progenitor to the present biodiversity of chickens in the
world.Tixier-Boichardet.al (2009) worked on characterization and monitoring of
indigenouspoultry genetic resources. They clearly depicted the characterization protocols on
thebasis of population size and structure, geographical distribution, production systems
inwhich the breed is found, phenotypic attributes (physical features, performance levelsand
any unique features), historical development of the breed (crossbreeding, selection). For
village chickens, information on specific features such as morphological diversity,
scavenging behavior, product quality and disease resistance along with molecular markers
contribute to define breed identity and can assign individuals to their population of origin and
proposed indicators for between- as well as within- population variability. Krygeret.al (2010)
described the opportunities and limitations of smallholder poultry production and overview of
current knowledge regarding the chicken genetic resources kept in smallholder production
systems in developing countries. FAO has classified poultry production into four sectors:
sectors 1 and 2 comprise large-scale commercial operations; sector 3 comprises small-scale
commercial farms; sector 4 comprises backyard and scavenging production, and is largely
based on indigenous birds, often belonging to local breeds that have been kept for many
years in a particular area and are adapted to local conditions. In a large numbers of
developing countries in Africa and Asia, indigenous birds constitute up to 80 percent of the
standing poultry population. The article provided FAO guideline for assessment of poultry
genetic resources
worldwide. Das et.al (2013) conducted study on 60000 Rhode Island Red (RIR) and
indigenous chickens reared at backyard poultry production system under scavenging at five
agro climatic zones in West Bengal, India. They studied twenty one variables under three
major criteria viz. physiological, economical and adoption of technology. The best possible
combination of input and supportive factor for economic backyard poultry farming at
farmers’ doorstep of various agro-climatic zones were also analyzed.Motaet.al (2001)
experimented with 250 apparently healthy Gallus gallusdomesticusfowl and found the mean
values for the WBC, heterophil, eosinophil, basophil,lymphocyte and monocyte, counts were
23.620, 7.690, 0.650, 1080,13.560 and 0.640cells/ μl respectively. Puthpongsiripornet.al
(2001) showed vitamin E supplementation in laying at 30 – 35 week age reduced the negative
effects of heat stress with improved in vitro proliferation of lymphocytes. Iheukwumere and
Herbert (2003) investigated the haematology and serum biochemistry of broiler chickens and
recorded the haematological parameters like PCV, RBC, WBC and Hb as 38.00%, 9.20 x
106/cu mm , 6.80 x 103/cu mm and 13.00 g /100 ml respectively. Zulkifliet.al (2003) studied
the effects of early age feed restriction and heat conditioning on heterophil/lymphocyte ratios
(HLR), heat shock protein (hsp)-70 expression and body temperature of heat stressed male
broiler chickens. The HLR response reduced to the heat challenge and hsp-70 response
improved both in feed restriction upto 60% and feed restriction with heat stress. They also
showed that in latter the birds exposed to heat became more hyperthermic than
normal.Mashalyet.al (2004) showed that during heat stress there was increase in HLR from
0.39 (normal) to 0.56 (heat stress) in 31 wk laying hen with peak production.
Use of genomic resources – uncovering important sequences

The chick genome has fewer, smaller genes and fewer pseudogenes than Xenopuslaevis,
zebrafish, mouse or human.This not only makes it a good anchor for the analysis of other
genomes but it also makes it simpler than in other species to detect true homologues of
developmentally important genes and to manipulate their expression (see transgenesis
below). The increase in available chick sequence and the completion of the genome has
allowed the search for novel members of families of genes which are important in
development. For example, chicken Tbx18 was initially discovered through BLASTing the
Tbx motif against the UMIST EST collection and has been since found to be important in
somite, limb and heart development (Tanaka and Tickle,2003). Comparison of the chicken
genome with other genomes has also revealed conserved regions of the genome that act as

regulatory elements for gene expression in specific spatio- temporal patterns during
development. Beckers et al. (1996) compared the sequence of the Hoxd-11 loci in chicken,
mouse and zebrafish sequence and uncovered one conserved region which controlled the
expression of Hoxd-11 during development, when tested in the mouse limb. In comparison
Lin et al. (2006) compared the regulatory sequences of chicken genes which showed a
transcriptional response to Shh in a microarray expression profile of chicken feather bud
development. They found in these genes a common transcriptional responsive element which
is also present in mouse and human and which was then found to be required for expression
of the targets of the Hh signalling pathway in a chicken cell line. And yet other groups have
combined the identification of chicken regulatory sequences with transgenesis in order to
dissect the spatio-temporal action of different enhancers during development (Uchikawa et
al., 2003; see transgenesis below). Furthermore it has only been realised since the large scale
sequencing of the developmental specific EST libraries that there are a large (as yet
undetermined) number of microRNAs (miRNAs) which have developmentally specific
expression patterns (Hubbard et al., 2005). miRNAs regulate gene expression at the level of
the transcript and the chick has already been instrumental in determining how they act in
development,for example that miR-196 regulates expression of Shhand Hoxb8 (Hornstein et
al., 2005) and that miR-206 is regulated by Fgf signalling in myogenesis (Sweetman et al.,
2006). The chick can also be used to investigate the expression patterns of miRNAs during
development through the use of LNA-modified probes (Darnell et al., 2006; Sweetman et al.,
2006). Furthermore advances in identifying miRNAs have opened up further potentials for
knocking down gene expression in the chick .
One dilemma in comparing embryo development between poultry species is the absence of a
common reference of sequential stages of morphogenetic development. To counter this
problem, the normal table of embryogenesis was devised to accurately assess embryo
development during the oviductal and incubation phases of embryogenesis . It is not only
important for research on the morphogenetic development of the avian embryo, but it is also
useful for investigators in the poultry industry attempting to uncover the basis of fertility and
hatchability problems The ability to differentiate a live from a dead embryo or a fertilized
from an unfertilized germinal disc (GD) is a prerequisite for determining whether the
problem lies with fertility or embryo mortality. Unfortunately, the ability of the hatchery
technicians and poultry scientists to differentiate a viable embryo from an early dead embryo
from an unfertilized GD is questionable . This differentiation is very important when
performing fresh egg breakouts or candling breakouts. When eggs are candled, the clear eggs
(i.e., those with no indication of embryo development) are all classified as unfertilized, and
the percentage of fertilized eggs gives what is known as candling fertility. When the clear
eggs are opened and examined (breakout) and the GD visually examined to differentiate an
early dead embryo from an unfertilized GD, then true fertility can be established . This
practice requires trained technicians and, in the case of eggs examined within 24 h from the
onset of incubation, a normal table as a reference to identify the actual stage of embryo
development. The usefulness of clear-egg breakouts in the turkey industry was demonstrated
by Krueger . Krueger’s data revealed the magnitude of early embryonic mortality of some
commercial turkey breeder flocks, suggesting that early embryonic mortality can be an
insidious problem. Precision in identifying the status (fertilized vs. unfertilized and stage of
embryonic development) of the GD is crucial in understanding the basis of hatch failures. In
studies describing the effect of preincubation on long-term storage of turkey and broiler eggs,
Fasenko et al. observed early embryo mortalities of 7.2% in turkeys and 6.2% in broiler
chickens. BakstnandAkuffo reported that when the GD is critically examined at breakout,
eggs from 18 turkey hens had 100% true fertility, 17 hens had 1 or more unfertilized or early
dead embryos, and, of the 10 hens producing 1 or more early dead embryos, 3 hens were
responsible for 50% of the early deads. Notwithstanding their commercial importance,
reports addressing early embryonic mortality and hatchability in ducks, geese, Japanese quail,
and Guinea fowl are limited. Brun et al. found no differences in early embryo mortality (2 to
3%) between mule (a commercial cross between a male Muscovy and female Pekin), Pekin,
and Muscovy embryos at candling after 6 d of incubation. However, breaking out the clear
eggs revealed an early embryonic mortality of 14.9% in the mules and 10.3% in the Pekin
and Muscovy eggs. More than 50 billion chickens are raised annually as a source of food, for
both their meat eggs. Chickens raisedfor eggs are usually called layers while chickens raised
for meat are often called broilers . On average a layinghen produces one egg/day. All laying
hens start to lay exactly when they are 21 weeks old. Planning istherefore required for egg
production to be constant so as to meet market demand. In areas where the climate ishot and
humid, commercial hybrid laying hens produce on average between 180 and 200 eggs/year.
In more temperate climates laying hen can produce on average between 250 and 300
eggs/year.Wild chickens meet their needs naturally, however chicken domestication has
introduced new environments and restrictions. Management systems have evolved and
become extensive. Confinement facilitates egg collection, chicken capture, reproduction,
protection from predators/pests and biosecurity measures. Chickens that are partially or

totally confined, require man to provide those needs which cannot be accommodated
naturally such as: provision of water and feed; shelter and/or ventilation; artificial lighting;
cleaning and/or disinfection of facilities. These needs become more intensive and critical as
the space/hen is reduced and production increases. Confinement systems include: on range in
fields/paddocks where climate permits; confined indoors in pens with access to range;
confined totally indoors either in pens or cages with either single or multiple hens/cage. The
relative performance of hens from 18 to 76 weeks of age in conventional cages, in enriched
cages and in pens revealed only small differences in egg production, mortality and feed/egg .
Globally, a range of relevant management systems is applied. The specific type is determined
largely by climatic and economic restraints. Modern poultry production occurs primarily in
enclosed buildings to protect the birds from weather, predators, and the spread of diseases
from wild birds. This has allowed farmers to greatly increase production efficiency while
significantly reducing the amount of labor required. The majority of egg production is carried
out using a conventional cage system, where layers live in cages and have limited mobility.
Housing density and reduced space per hen have aroused consumer concern about hen
welfare. In this perspective management systems have developed to meet the chicken’s needs
for health and safety. These usually include: provision of water and feed; shelter and/or
ventilation; artificial lighting; cleaning and/or disinfection of facilities. This also happened in
response to worldwide research and development activities about welfare-friendly poultry
housing system and husbandry as well as in compliance to a European Union Council
Directive (1999/74/EC) on welfare of laying hens. This directive requires conventional laying
cages to be phased out by 2012. Traditional cages have been modified and “enriched” cages
and aviaries have been developed and encouraged along with free- range systems. The
benefits of free-range poultry farming for laying hens include opportunities for natural
behaviors such as pecking, scratching, foraging and exercise outdoors . However the
production costs of these modified measures are much higher than those of eggs from caged
hens . Layer operation size in developed countries has increased dramatically during the last
century. Flocks of 100,000 laying hens are common with some exceeding 1 million . Large
layer farms consist of several large houses of in-line and off-line types . In in-line production
systems, eggs from all houses are gathered and transferred to an adjacent plant for processing
and refrigeration prior to shipping. The appropriate nutrition provision to hen is pre-requisite
for optimal egg production. Nutritionists design rations to meet the laying hen’s energy,
protein, minerals and vitamins requirements determined by maintenance, body weight and

level of egg production. Ingredient selection is determined by availability, price and, if
available, nutrient biological availability estimates, to minimize cost of the ration. Specific
fats and cereal grains provide energy, content and digestibility of essential amino acids
determine the protein source, a variety of vitamins and trace minerals are included in
supplements given to egg laying hens. The laying hen requires higher levels of calcium and
vitamins A, D and choline than other chicken. Hens can be sustained on diets with limited
nutrients; however, production levels will be reduced accordingly. To meet consumer
demands, “organic eggs” are now produced. Hens producing organic eggs has specific
housing requirements as outlined by the European Commission Regulation (EC) No
889/2008. This poultry farming must be kept in a free range system and fed only organic feed
free of antibiotics and synthetic fertilizers. Hen’s feed must include grains from only crops
certified as “organic”. Antibiotics can be used but only for the control disease outbreaks.
Laying hens must have free access to the outdoors with adequate vegetation during the day
time .Family poultry production has provided opportunities for income generation to the local
communities, apart from satisfying their own needs. This production system is promoted in
Ghana, Tanzania, and Zambia by the Food and Agriculture Organization of the United
Nations and the International Egg Commission. Recommendations to improve management
and production for these smaller and indigenous operations are made available .Worldwide
active research activities are on record with the objective to improve poultry practices with
emphasis on housing systems . In the same perspectives symposia, with the intervals of four
years, were periodically held in number of European countries (Denmark, 1981; Germany,
1985; France, 1989; UK, 1993;Netherland, 1997; Switzerland 2001; Poland, 2005 and Italy,
2009) to address the poultry farming systems and related issues. Latest and updated research
based contributions on this subject gathered at the 9th European symposium on poultry
welfare held in Uppsala, Sweden . This meeting illustrated extensive research efforts made in
number of countries by evaluating welfare of hens including housing and husbandry research
and development to disseminate information all across the world.

MATERIALS AND METHODS
The chicken (taxon -Gallus gallus) embryo develops and hatches in 20 to 21 days and has
been extensively used in embryology studies. Historically, the chicken embryo was one of the
first embryos studied, readily available and easy to incubate, embryo development can be
directly observed by cutting a small window in the egg shell. A key to this model organism
study was the establishment of a staging atlas by Hamburger & Hamilton in 1951, which
allowed specific developmental landmarks to be seen and correlated with experimental
manipulations of development. This much cited paper included images of all key stages and
was more recently republished in the journal Developmental Dynamics (1993), for a new
generation of avian researchers. Probably just as important has been the recent chicken
genome sequencing, providing a resource to extend our knowledge of this excellent
developmental model. Fertilized eggs can be easily maintained in humidified incubators and
during early stages of development the embryo floats on to of the egg yolk that it is using for
nutrition. As the embryo grows it sinks into, or below the, yolk. The regular appearance of
somites allowed early experimenters to accurately stage the embryo. The embryo was
accessible and easy to manipulate (limb grafts/removal etc) that were informative about
developmental processes. Chicken cells and tissues (neural ganglia/fragments) are also easy
to grow in tissue culture. The discovery that quail cells have a different nuclear appearance
meant that transplanted cells (chick/quail chimeras) could be tracked during development.
For example, LeDourian's studies showed how neural crest cells migrate widely throughout
the embryo.
Principles Of Incubation
Five major functions are involved in the incubation and hatching of poultry eggs. They are:
 Temperature
 Humidity
 Ventilation (Oxygen and Carbon dioxide level and air velocity)
 Position of eggs
 Turning of eggs

1. TEMPERATURE
Temperature is the most critical environmental concern during incubation because the
developing embryo can only withstand small fluctuations during the period. Embryo starts
developing when the temperature exceeds the Physiological Zero. Physiological zero is the
temperature below which embryonic growth is arrested and above which it is reinitiated.
The physiological zero for chicken eggs is about 75oF (24oC). The optimum temperature for
chicken egg in the setter (for first 18 days) ranges from 99.50 to 99.75 o Fand in the hatcher
(last 3 days) is 98.50 F. Relative humidity should be set at 55 to 60 percent. If the incubator
uses a passive humidity control system, add water to the pan or trough daily to maintain
correct humidity levels. If the humidity in the incubator is too low or too high, the hatch will
fail.
Insufficient humidity causes:
◆The air cell to be too large at the time of hatch
◆The contents of the egg be too viscous for the chick to turn
◆The membranes to be too tough to break
◆The navel to not close properly
Excess humidity will causes:
◆ Too little water to evaporate from the egg
◆The air cell to be too small for the chick to reach during the hatching process
◆The chick to drown or be too swollen with water to turn in the egg
◆The yolk sac to be too large for the navel to close completely.
The air cell of the egg should become larger as incubation progresses because of the balance
between temperature and humidity during incubation. Chicken eggs lose 12 percent to 14
percent of their total weight to evaporation during incubation. You can weigh racks of eggs
during incubation to detect problems with humidity and evaporative loss before a hatch is
destroyed. The chick embryo uses oxygen and produces carbon dioxide. This gas exchange is
insignificant during early incubation or when a small number of eggs are incubated; however,
follow the manufacturer’s recommendations to assure that developing chicks have adequate
oxygen available. Near the end of the incubation period, the shell nearly filled with the
embryo and a full incubator requires large amounts of oxygen. Ensure adequate ventilation
and monitor wet and dry bulb temperatures very carefully during the last third of incubation.
2. HUMIDITY
Incubation humidity determines the rate of moisture loss from eggs during incubation. In
general, the humidity is recorded as relative humidity by comparing the temperatures
recorded by wet-bulb and dry-bulb thermometers. Recommended incubation relative
humidity for the first 18 days ranging between 55 and 60% (in setter) and for the last 3 days
ranging between 65 and 75%. Higher humidity during hatching period is given to avoid
dehydration of chicks.
3. VENTILATION
Ventilation is important in incubators and hatchers because fresh oxygenated air is needed for
the respiration (oxygen intake and carbon dioxide given off) of developing embryos from egg
setting until chick removal from the incubator.
The oxygen needs are small during the first few days compared to the latter stages of
development. Oxygen content of the air at sea level is about 21%. Generally the oxygen
content of the air in the setter remains at about 21%. For every 1% drop in oxygen there is
5% reduction in hatchability. Carbon dioxide is a natural by-product of metabolic processes
during embryonic development and is released through the shell. The tolerance level of CO2
for the first 4 days in the setter is 0.3%. CO2 levels above 0.5% in the setter reduce
hatchability and completely lethal at 5.0%.
Since the normal oxygen and CO2 concentrations present in air seem to represent an
optimum gaseous environment for incubating eggs, no special provision to control these
gases is necessary other than to maintain adequate circulation of fresh air at the proper
temperature and humidity.
Position of eggsartificially incubating eggs should be held with their large ends up.
It is natural for the head of the chick to develop in the large end of the egg near the air cell,
and for the developing embryo to orient itself so that the head is uppermost. When the eggs
are incubated with the small end up, about 60% of the embryos will develop with the head
near the small end. Thus, when the chick is ready to hatch, its beak cannot break into the air

cell to initiate pulmonary respiration. Eggs positioned horizontally will incubate and hatch
normally as long as they are turned frequently. Under normal circumstances eggs are set with
large end up for the first 18 days (in setter) and in horizontal position for the last 3 days (in
hatcher).
5. Turning of eggs
Birds, including chickens and quail, turn their eggs during nest incubation. Nature provides
nesting birds with the instinct of turning eggs during incubation. Similarly eggs to be turned
at least 8 times a day. Turning of eggs during incubation prevents the developing embryo
adhering to the extra-embryonic membranes and reduces the possibility of embryo mortality.
In large commercial incubators the eggs are turned automatically each hour i.e. 24 times a
day. Most eggs are turned to a position of 45o from vertical, and then reversed in the opposite
direction to 45o from vertical. Rotation less than 45o are not adequate to achieve high
hatchability. Turning is not required in Hatcher.
Factors Setter Hatcher

Temperature 99.50 to 99.75 F 98.50 F
Relative humidity 55-60% 65-70 %
Position Large end up Horizontal
Turning Manual 8 times automatic 24 Not running
times
Handling of Hatching Eggs and Storage
Table 2.2 Effects of factors involve in hatching and storage of eggs
The quality of hatching egg cannot be improved after lay but one can reduce the loss in
hatching egg quality by adopting some standard procedures. Maintaining egg quality in the
breeder houseThe hen will lay her eggs over nesting material. Use of enough clean, dry and
mold-free nesting material can avoid cracked and dirty eggs. Similarly hens to be trained to
use nests to lay eggs instead of laying on floors by providing sufficient number of nest boxes
well in advance before the laying starts.The frequency of hatching egg collection is very
important to maintain quality. Hatching eggs should be collected at least 4 times a day.
Hatching eggs are susceptible to contamination and every effort must be made to reduce the
microbial load. Therefore, it is imperative that people wash and sanitize their hands before
collecting eggs from the nests or egg belts. The flats that eggs are placed on must also be
sanitized and free of organic material.
Selection of hatching eggs

Not all eggs laid by a breeding flock are set. Eggs that are cracked, dirty or misshapen are
usually not used for hatching. Very small or very large eggs do not hatch as well as eggs in
the middle size range.Eggs with thin or very porous shells are not likely to hatch well because
of excessive losses of water during incubation .
Reducing contamination of hatching eggs
Poor hatching egg sanitation can be a major cause of lower hatchability and poor chick
quality. There is no such thing as a sterile eggshell. More bacteria are picked up on the shell
when the egg passes through the cloaca where urine and intestinal contents also pass. The
bacterial load found on an eggshell at the time of lay ranges from 300 to 500 organisms.
After oviposition, every surface the egg comes in contact with can further inoculate the shell
surface. After an egg is laid it begins to cool. During the cooling process the egg contents
begin to shrink and producing negative pressure. This is one of the more opportune times for
bacteria on the shell surface to penetrate the eggshell. Egg has many natural defense
mechanisms to reduce bacterial penetration. The shell itself provides some protection. The
cuticle on the surface of the eggshell is the best natural barrier to penetration. The inner and
outer shell membranes provide additional barriers. The albumen provides a somewhat
effective control over contamination. The albumen has a high pH in which most bacteria
cannot survive. The chalazae contain an enzyme, lysozyme, which has antibacterial
properties.Many breeder people choose some methods to reduce the microbial load over the
eggshell. Sanding, buffing, and wiping the hatching eggs are not good methods of sanitation.
Sanding and buffing will remove at least part of the cuticle resulting in eggs that are more
susceptible to penetration. Fumigation with formaldehyde gas is an effective method of
sanitizing hatching eggs. Solutions containing quaternary ammonium compounds, formalin,
hydrogen peroxide or phenols may be moderately effective in reducing the microbial load
over hatching eggs. DO NOT wash eggs unless necessary. If it is necessary to wash eggs
always use a damp cloth with water warmer than the egg. This causes the egg to sweat the
dirt out of the pores. Never use water cooler than the egg. Also, do not soak the eggs in
water. In the present study, the chicken eggs which belong to same date of hatch were
utilized. Three numbers of eggs were collected every day and set for incubation. At the end of
the 20th day, all the eggs were opened and day wise embryos were collected from day 1 to
day 20. The eggs were cracked into finger bowl which contained 0.9% warm saline solution

without disturbing the yolk. With a fine forceps and scissors, theblastoderm were separated
from the yolk and allowed to float away in the saline solution. By using a microscopic slide,
the embryos were picked up and transferred to a separate bowl containing fresh saline
solution upto fifth day embryo stage. The remaining stage of chick embryos were collected
by using forceps and scissors and were transferred to warm saline solution. Afterwards,
excess saline from the dish were pipetted out and they were replaced with fixative solution
(Neutral buffered formalin and Bouin’s fluid). The embryos were allowed for overnight
fixation and were used to study the different morphological development (Figs. 1 and 2)
(Schoenwolf and Watterson, 1989).
Materials
• Chick Ringer’s solution
• Chicken eggs
• 1 Petri dish
• Forceps
• Scissors for dissection
• Methylene Blue
• Permanent mounted embryo tissue (33hours, 72 hours and/or 96 hours)
Procedure
1. Crack the egg into a Petri dish filled with Ringer’s solution.
2. The egg will turn so that the embryo is on top; if not, twist the chalazae (the thick, twisted
strands of the albumen) until the embryo is on top.
3. Make a cut in the yolk outside at the lower side of the blastoderm.
4. Grasp the opaque peripheral area of the blastoderm (which is continuous with the yolk)
with your forceps.
5. Completely cut around the blastoderm while holding it with your forceps and gently pull
the blastoderm loose, moving it to a part of the dish with no yolk or albumen.

6. Add 2-3 drops of methylene blue and allow it to sit for 5 minutes. This is done to enhance
contrast.
7. Rinse the embryo with warm Ringer’s solution.
8. Time the heart beat of the embryo: how many beats per minute do you observe? Do this 5
times for every 5 minutes interval.
9. Make a rough sketch of the isolated chick embryo and take observations.
10. Measure and record the size of the embryo.
11. Compare and contrast permanent mounted embryo tissue with the live embryo
RESULT DISCUSSION AND FUTURE WORK

The first day of chick development takes place inside the mother hen (in utero), during which
the embryo progresses from fertilization to late blastula/early gastrula formation. The salient
features of developmental anatomy in this period are conserved among the sauropsids (birds
and reptiles). Many of these features are also shared in prototherian (monotreme) embryos,
whereas metatherian (marsupial) and eutherian (placental) embryos display significant
variations. Important for understanding the evolution of early development in amniotes, the
knowledge of cellular and molecular mechanisms regulating in utero chick development may
also offer valuable insight into early lineage specification in prototherians and conserved
features in mammalian early development. This commentary provides a snapshot of what is
currently known about intrauterine chick development and identifies key issues that await
further clarification, including the process of cellularization, allocation of maternal
determinants, zygotic gene activation, mid-blastula transition, cell layer increase and
reduction, radial symmetry breaking, early lineage segregation, and role of yolk syncytium in
early patterning
Day 1
On 24 hours of incubation, the first indication of development of body plan, the head fold was noticed
at the cranial end. It appeared like a crescent shaped structure. The neural folds were visible which
expanded posteriorly from the head fold. At this stage, pairs of somites on either side of notochord
and primary optic vesicle were noticed (Bellairs and Osmond, 2005).
Day 2
The cranial flexure and trunk flexure were started to visible which were the indications of external
appearance of chick. Somites and amniotic vesicle were clearly observed. (Pikalow et al., 1994)
Day 3
On day 3, the cervical flexure formed an acute angle with the axis of the trunk. The trunkflexure was
entirely disappeared and the trunk was in a straight line with the base of the tail (Hamburger and
Hamilton, 1951). Distinct vitelline blood vessels on the yolk and to the embryo were clearly seen and
also the heart pumping and circulation were distinctly noticed to the naked eye (De Haan, 1990)
Day 4
On day 4, both wing and limb bud were noticed as a bulge at the cranial and caudal regions
respectively. At this stage, the head bends round in the neck region and it came to lie at right angles to
the trunk (Flynn et al., 1991). Faint eye pigmentation was noticed.
Day 5
On day 5, embryo showed distinct optic vesicles with eye pigmentation on the lateral aspects of
cephalic region and also prosencephalic vesicle at the head region. In the wing demarcation of elbow
and knee joints were observed. The digital plate in wing was distinct and first three toes were
separated (Murray and Wilson, 1994).
Day 6
As mentioned by Hamilton (1965), distinct outgrowth of beak was visible and neck between collar
and mandible had lengthened. The external auditory meatus was noticed.
Day 7
On seventh day, three major segments of leg and wing are clearly demarcated. The gap
between beak and mandible had narrowed. Feather germs began to appear at brachial region
and at the level of leg region. Scleral papilla was clearly visualised (Bellairs and Osmond,
2005).
Day 8
On eighth day, the embryo showed well distinct digits and toes and they were lengthened. The
mandible and neck also lengthened. The egg tooth was clearly viewed. The feather germs became
clearly visible and the scleral papilla formed almost a complete circle (Bellairs and Osmond, 2005).
Day 9
In the limbs, grooves appeared between first, second, third and fourth digits and which later
forms the webs of digits. Distinct phalanges in toes were noticed. Eyelids began to overgrow
the eyeball and eyelid circumference becomes ellipsoidal (Majumdar, 1988).
Day 10
In the tenth day of embryological development, distal segment of wings and legs became longer and
primordia of claws were visible. The nostril was narrowed to a slit like opening and labial groove was
clearly visible on the tip of the upper jaw (Bellairs and Osmond, 2005). As mentioned by Hamilton
(1965), the flight feathers and nine rows of feather germs between upper eyelid and dorsal midline
were not seen and comb primordia were also not noticed.
Day 11

On eleventh day, onset of cornification of claws was seen and pads on plantar surface of foot were
visible. In the leg, scales were seen over the entire surface of leg. The openingbetween the upper and
lower eyelids was much reduced. The feather germs became conical and noticed over many areas of
the body especially dorsal midline and caudal regions of body. Distinct development of flight feathers
were noticed (Hamilton, 1965).
From eighth to twelfth day, the development of eyelids, feather germs, growth of legs, wing, beak and
feathers were gradually increased. Afterwards, there was only growth of structures which were
already formed was noticed up to 20th day. There was no new development of structures were seen
after eighth day. Only growth of organs and structures which increased the weight of embryo was
observed. The study on sequential development of chick embryos in Namakkal variety of chicken
concluded that the development of each and every parts of body were gradually started at 24 hours of
incubation and most of the structures were fully developed at eighth to twelfth day of incubation.
Afterwards, only increase in growth of structures was observed.

Fig 1.16:photograph showing the sequential stages of chick embryos from day 4 to day 12
Fig 1.17: figure showing the sequential stages of chick embryos from 13 day to day 19

References
1. Bellairs. R and M. Osmond. 2005. The Atlas of Chick Development. Second edition,
Elsevier publication, California.
2. De Hann, R.L. 1990. The embryonic origin of the heartbeat. In The Heart 7th ed.
McGraw-Hill, New York, pp.72-77
3. Flynn, M.E., Pikalow, A.S., Kimmelman, R.S. and Searls, R.L. 1991. The mechanism of
cervical flexure formation in the chick. Anat. Embryol. 184: 411-20.
4. Hamburger, V and Hamilton, H.L. 1951. A series of normal stages in the development of the
chick embryo. J. Morph. 88: 49-92
5. Hamilton, H.L. 1965. Lillies Development of the Chick, an Introduction to Embryology Holt
Rinehart and Winston, New York.
6. Horder,T.J., Witowski, J.A. and Wylie, C.C. 1986. A History of Embryology. Cambridge
University Press.
7. Majumdar, N.N. 1988. Textbook of Vertebrate Embryology. Tata McGraw-Hill, New Delhi.
8. Murray, B.M. and Wilson, D.J. 1994. A scanning electron microscopic study of the normal
development of the chick wing from stages 19-36. Anat. Embryol. 89: 147-58.
9. Pikalow, A.S., Flynn, M.E. and Searls, R.L. 1994. Development of the cranial flexure and
Rathke’s pouch in the chick embryo. Anat. Rec. 238: 407-14.
10. Schoenwolf, G.C. and Watterson, R.L. 1989. Laboratory Studies of Chick, Pig and Frog
embryos. Guide and Atlas of Vertebrate Embryology, 6th edition. Macmillan, New York.
11. Bellairs, R., and M. Osmond. 1998. The Atlas of Chick Development. Acad. Press, New
York, NY.
12. Bakst, M. R., and V. Akuffo. 2002. Impact of egg storage on embryo development. Avian
Poult. Biol. Rev 13:125–131.
13. Bakst, M. R., S. K. Gupta, W. Potts, and V. Akuffo. 1998. Gross appearance of the turkey
blastoderm at oviposition. Poult. Sci. 77:1228–1233.
14. Krueger, K. K. 1990. Fertility in female turkeys: How to manage it? Pages 205–212 in
Control of Fertility in Domestic Birds.J. P. Brillard, ed. INRA, Nouzilly, France.
15. Fasenko, G. M., V. L. Christensen, M. J. Wineland, and J. N. Petitte. 2001. Examining the
effects of prestorage incubation of turkey breeder eggs on embryonic development and
hatchability of eggs stored for four or fourteen days. Poult. Sci. 80:132–138.
16. Fasenko, G. M., F. E. Robinson, A. I.Whelan, K.M. Kremeniuk, and J. A. Walker. 2001.
Prestorage incubation of long-term stored broiler breeder eggs: 1. Effects on hatchability.
Poult. Sci. 80:1406–1411.
17. Brun, J. M., N. Sellier, M. M. Richard, F. Batellier, and J. P. Brillard. 2004. Effect of in vivo
sperm storage and breeder hen’s age on fertility and embryonic survival in pure Pekin duck
hens.
18. and their crossbreeding with Muscovy drakes. Proc. XXII World’s Poult. Congr., Istanbul,
Turkey.
19. Hamburger, V., and H. L. Hamilton. 1951. A series of normal stages in the development of
the chick embryo. J. Morphol. 88:49–92.
20. Eyal-Giladi, H., and S. Kochav. 1976. From cleavage to primitive streak formation:
Complementary normal table and a new look at first stages of the development of the chick. I.
General morphology. Dev. Biol. 49:321–337.
21. Gupta, S. K., andM. R. Bakst. 1993. Turkey embryo staging from cleavage through hypoblast
formation. J. Morphol. 217:313–325.
22. Dupuy, V., B. Nersessian, and M. R. Bakst. 2002. Embryonic development from first
cleavage through seventy-two hours incubation in two strains of Pekin duck (Anas
platyrhynchos). Poult.Sci. 81:860–868.
23. Stepinska, U., and B. Olsanska. 1983. Cell multiplication and blastoderm development in
relation to egg envelope formation during uterine development of Japanese quail (Coturnix
coturnixjaponica) embryo. J. Exp. Zool. 228:505–510.
24. Bakst, M. R., S. Gupta, and V. Akuffo. 1997. Comparative development of the turkey and
chicken embryo from cleavage through hypoblast formation. Poult. Sci. 76:83–90.
25. Pekin ducks, Grimaud Fre`res Se´lection, Roussay, France.
26. Bakst, M. R. and H. Cecil. 1997. Techniques for Semen Evaluation, Semen Storage, and
Fertility Determination. Poult. Sci. Assoc., Savoy, IL.
27. Stemi SV11 stereomicroscope, Carl Zeiss SA, Le Pecq, France.
28. Photomicrographs were obtained with a 4.0 megapixel Powershot G3 Canon Digital Camera,
Canon SA, La Garenne Colombes, France.
29. Altabef M, Clarke JDW, Tickle C: Dorso-ventral ectodermal compartments and origin
of apical ectodermal ridge in developing chick limb. Development 124: 4547–4556
(1997).
30. Beckers J, Gerard M, Duboule D: Transgenic analysis of a potential Hoxd-11 limb

regulatory element, present in tetrapods and fish. Dev Biol 180: 543–553 (1996).
31. Bellairs R, Osmond M: The Atlas of Chick Development, 2 nd Ed (Elsevier
Academic Press, Oxford 2005).
32. Boardman P, Sanz-Ezquerro JJ, Overton I, Burt DW, Bosch E, et al: A comprehensive
collection of chicken cDNAs. Curr Biol 12: 1965–1969 (2002).
33. Creuzet S, Couly G, Le Douarin NM: Patterning the neural crest derivatives during
development of the vertebrate head: insights from avian studies. J Anat 207: 447–459
(2005).
34. Darnell DK, Simran K, Stanislaw S, Konieczka JK, Yatskievych TA, Antin PB:
MicroRNA expression during chick embryo development. Dev Dyn 235: 3156–3165
(2006).
35. Das RM, Van Hateren NJ, Howell GR, Farrell ER, Bangs FK, et al: A robust system
for RNA interference in the chicken using a modified microRNA operon. Dev Biol
294: 554–563 (2005).
36. Davey MG, Paton IR, Yin Y, Schmidt M, Bangs FK, et al: The chicken talpid 3 gene
encodes a novel protein essential for Hedgehog signaling. Genes Dev 20: 1365–1377
(2006).
37. Davey MG, James J, Paton IR, Burt DW, Tickle C: Analysis of talpid 3 and wild-type
chicken embryos reveals roles for Hedgehog in development of the limb bud
vasculature. Dev Biol 301: 155–165 (2007).
38. Duboule D, Dollé P: The structural and functional organization of the murine HOX
gene family resembles that of Drosophila homeotic genes. EMBO 8: 1497–1505
(1989).
39. Eblaghie MC, Lunn JS, Dickinson RJ, Munsterberg AE, Sanz-Ezquerro JJ, et al:
Negative feedback regulation of FGF signaling levels by Pyst1/ MKP3 in chick
embryos. Curr Biol 13: 1009– 1018 (2003).
40. Ede DA, Kelly WA: Developmental abnormalities in the trunk and limbs of the
talpid3 mutant of the fowl. J Embryol Exp Morphol 12: 339–356 (1964).
41. Eyal-Giladi H, Kochav S: From cleavage to primitive streak formation; a

complementary Normal table and a new look at the first stages of the development of
the chick. I. General morphology. Dev Biol 49: 321–337 (1975). Ferretti P, Tickle C:
The Limbs, in
42. Feretti P, Copp A, Tickle C, Moore G (eds): Embryos, Genes and Birth Defects, 2nd
Ed, pp 123–166 (Wiley, Chichester 2006).

Chick Embryology

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Chick Embryology

Uploaded by

Copyright:

Available Formats

INTRODUCTION

Embryology is the study of the development of organisms. This is as true of plants as it is of

1 | Development of chick embryo as a model system for embryological study of Vertebrate

2 | Development of chick embryo as a model system for embryological study of Vertebrate

3 | Development of chick embryo as a model system for embryological study of Vertebrate

4 | Development of chick embryo as a model system for embryological study of Vertebrate

5 | Development of chick embryo as a model system for embryological study of Vertebrate

GONADS & GAMETES

6 | Development of chick embryo as a model system for embryological study of Vertebrate

Fig 1.3-Figure showingreproductive organ of female hen

9 | Development of chick embryo as a model system for embryological study of Vertebrate

10 | Development of chick embryo as a model system for embryological study of Vertebrate

Fig 1.5-Figure showing reproductive tract of laying hen.

OVULATION & MATURATION:

11 | Development of chick embryo as a model system for embryological study of Vertebrate

12 | Development of chick embryo as a model system for embryological study of Vertebrate

FORMATION OF ACCESSORY LAYERS AROUND THE FERTILIZED

13 | Development of chick embryo as a model system for embryological study of Vertebrate

Fig:1.7-Figure showing parts of hens egg

14 | Development of chick embryo as a model system for embryological study of Vertebrate

Fig:1.8-Figure showing parts of shell membrane of egg

15 | Development of chick embryo as a model system for embryological study of Vertebrate

16 | Development of chick embryo as a model system for embryological study of Vertebrate

18 | Development of chick embryo as a model system for embryological study of Vertebrate

19 | Development of chick embryo as a model system for embryological study of Vertebrate

20 | Development of chick embryo as a model system for embryological study of Vertebrate

21 | Development of chick embryo as a model system for embryological study of Vertebrate

22 | Development of chick embryo as a model system for embryological study of Vertebrate

23 | Development of chick embryo as a model system for embryological study of Vertebrate

24 | Development of chick embryo as a model system for embryological study of Vertebrate

25 | Development of chick embryo as a model system for embryological study of Vertebrate

26 | Development of chick embryo as a model system for embryological study of Vertebrate

27 | Development of chick embryo as a model system for embryological study of Vertebrate

A. To make direct and daily visual observations of living embryos;

B. To learn the anatomy and physiology of the egg and embryos;

28 | Development of chick embryo as a model system for embryological study of Vertebrate

Classical experimental embryology

29 | Development of chick embryo as a model system for embryological study of Vertebrate

30 | Development of chick embryo as a model system for embryological study of Vertebrate

Tracing cell fate

31 | Development of chick embryo as a model system for embryological study of Vertebrate

Molecular basis of development

32 | Development of chick embryo as a model system for embryological study of Vertebrate

Genes and development

33 | Development of chick embryo as a model system for embryological study of Vertebrate

34 | Development of chick embryo as a model system for embryological study of Vertebrate

35 | Development of chick embryo as a model system for embryological study of Vertebrate

Use of genomic resources – uncovering important sequences

37 | Development of chick embryo as a model system for embryological study of Vertebrate

39 | Development of chick embryo as a model system for embryological study of Vertebrate

40 | Development of chick embryo as a model system for embryological study of Vertebrate

41 | Development of chick embryo as a model system for embryological study of Vertebrate

 Ventilation (Oxygen and Carbon dioxide level and air velocity)

42 | Development of chick embryo as a model system for embryological study of Vertebrate

Insufficient humidity causes:

◆The air cell to be too large at the time of hatch

◆The membranes to be too tough to break

◆The navel to not close properly

Excess humidity will causes:

◆ Too little water to evaporate from the egg

44 | Development of chick embryo as a model system for embryological study of Vertebrate

Factors Setter Hatcher

Selection of hatching eggs