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Genomics and phylogenetic lineage of different varieties of Indian
peafowl (Pavo cristatus) for pure-breeding in Pakistan.
JUNAID NASEER
2014-VA-772
A THESIS SUBMITTED IN THE PARTIAL FULFILLMENT OF
THE REQUIREMENTS FOR THE DEGREE
OF
DOCTOR OF PHILOSOPHY
IN
WILDLIFE AND ECOLOGY
UNIVERSITY OF VETERINARY & ANIMAL SCIENCES,
LAHORE
2017
To,
The Controller of Examinations,
University of Veterinary and Animal Sciences,
Lahore.
ii
We, the supervisory committee, certify that the contents and form of the
thesis, submitted by JUNAID NASEER, Regd. No. 2014-VA-772 have been found
satisfactory and recommend that it be processed for the evaluation by the External Examiner
for the award of Degree.
Dr. Khalid Mahmood Anjum _________________________
Supervisor
Dr. Waseem Ahmad Khan ________________________
Member
Dr. Muhammad Imran ________________________
Member
iii
The best human being on earth
For their kind support
DEDICATIONS
Holy Prophet Hazrat Muhammad
(Peace be upon him)
My Parents
My Teachers and Friends
iv
TABLE OF CONTENTS
DEDICATION………………………………………………………………………..... i
ACKNOWLEDGEMENTS ………………………………………………………….. ii
LIST OF TABLES …………………………………………………………………..... iii
LIST OF GRAPHS…………………………………………………………………….. v
LIST OF FIGURES…………………………………………………………………….. vi
Sr. No. Chapters Page No.
1. INTRODUCTION 01
2. REVIEW OF LITERATURE 05
3. MATERIALS AND METHODS 23
4. RESULTS 29
5. DISCUSSION 81
6. SUMMARY 97
7. LITERATURE CITED 99
8. ANNEXURE 106
v
Acknowledgements
All praises for Almighty Allah, the creator of the universe. The most compassionate and
merciful, who inspite of numerous difficulties capacitated me to accomplish the present
research work. All my love for Holy Prophet (Peace Be Upon Him), the messenger of Allah.
I would like to express my sincere gratitude to my advisor Prof. Dr. Khalid Mahmood
Anjum for the continuous support of my Ph.D study and related research, for his patience,
motivation, and immense knowledge. His guidance helped me in all the time of research and
writing of this thesis. I could not have imagined having a better advisor and mentor for my
Ph.D study. Besides my advisor, I would like to thank the rest of my thesis committee: Dr.
Waseem Ahmad khan and Dr. Muhammad Imran for their insightful comments and
encouragement, but also for the hard question which incented me to widen my research from
various perspectives. My sincere thanks also goes to Dr. Muhammad Shoaib, Dr. Asif
Munir, and Dr. Muhammad Awais Nazir, who provided me an opportunity to join their
team as intern, and who gave access to the laboratory and research facilities. Without they
precious support it would not be possible to conduct this research. I apologize if I have
caused anger or offence to anybody, and the errors that remain in the manuscript. With
tremendous pleasure I feel deep gratitude for my very yanki friends Muhammad Awais
Nazir, Azeem Ullah Khan, Syad Ghulam Abbas, Waqas Ali, Ali Hassan Badine, Zaib
Ali, Ahsan Zafar, Jahanzab Sarwar, Ali Yasir, Ali Asghar, Rooh Ullah and Dr. Ahsan
Anjum for their awesome company. May Allah bless all these people with long, happy and
peaceful lives (Amen). Thanks to all those who taught me every single word in my life
and who loved me.
Last but not the least, I would like to thank my family: my parents and to my brothers and
sister for supporting me spiritually throughout writing this thesis and my life in general.
JUNAID NASEER
1
LIST OF TABLES
Table Title Page No.
3.1 Components of the illustraTM Nucleon BACC2
RPN850)
24
3.2 Universal Primer sequences for cytochrome b
amplification (Verma and Singh, 2003)
27
3.3 PCR reaction mixture for cytochrome b amplification 27
3.4 Thermo cycler conditions for cytochrome b
amplification (Gupta, et al., 2005)
28
4.1 Frequency of Mating of Female Pavo cristatus 29
4.2 Number of Time Copulation of Female Pavo cristatus 31
4.3 Number of Mates of Female Pavo cristatus 32
4.4 Percentage of time (hours) allocated for each behavioral
activity of Indian peafowl for a period of six weeks
(breeding season)
33
4.5 Average Number of Eggs of Pavo cristatus 34
4.6 Breeding Season of Pavo cristatus 36
4.7 Range Time to Hatching of Pavo cristatus 38
4.8 Breeding Interval of Pavo cristatus 40
4.9 Age at Reproductive Maturity of Female Pavo cristatus 42
4.10 Age at Reproductive Maturity of Male Pavo cristatus 44
4.11 Nucleotide Sequence of Pavo cristatus mitochondrial
cytochrome b gene (1-1143bp)
60
4.12 Homology % of different sequence of mitochondrial 78
Introduction
2
cytochrome b with reference sequence Accession No.
L08379.1
4.13 Genetic Homology Percentage between Pavo cristatus
at Government Zoological Gardens and Private Sectors
78
Introduction
3
LIST OF FIGURES
Figure Title Page No.
4.1 Graphical representation of frequency of mating of
female Pavo cristatus
30
4.2 Average clutch size of Pavo cristatus at both
government zoological gardens and private sectors
35
4.3 Breeding behaviour of Pavo cristatus in different
months
37
4.4 Hatching Time of Pavo cristatus at different locations 39
4.5 Breeding Interval of Indian Peafowl at different
locations
41
4.6 Age at Reproductive Maturity of Female Indian
Peafowl
43
4.7 Age at Reproductive Maturity of Male Indian Peafowl 45
4.8 Genome Sequence Variation between Indian Peafowl at
Government Zoological Gardens and Private Sectors
79
4.9 The PCR product photograph of samples from govt.
zoological gardens and private sector with standard
marker (M).
79
4.10 Phylogenetic tree of the sequence obtained (This study)
with reference sequence available in NCBI with
accession no. of L08379.1.
80
Introduction
4
CHAPTER 1
INTRODUCTION
Since stone age, aves are thought to have close relation with human beings as their glimpses
were engraved on caves in Spain which are supposed to be 25,000 years old (Harikrishnan et
al. 2010). Birds are beautiful natural resources also beneficial for human beings in several
ways such as economic returns, environmental controller, meat production, bio-indicators,
source of knowledge and for recreation purposes (Araki et al. 1989). Pheasants and Indian
peafowl have more than 180 species worldwide which belong to order Galliformes and class
Aves. In developed regions of the world, e.g. shooting of game birds is the most industrialized
and productive business (Gupta et al. 2005).
The Indian peafowl is most beautiful, royal and big bird with long and colorful
feathers. Among all the pheasants it is considered the most beautiful and handsome pheasant
widely in the world. The Indian peafowl has body length 92-122 cms wingspan 240 cms,
train 140-160 cms and weight up to 2.75 Kgs (Ramesh and McGowan 2009). The distribution
of Indian, blue or common Indian peafowl (Pavo cristatus), is mainly confined to the Indian
subcontinent; southern part of the Himalayas, across Pakistan, eastern part of the Indus river
valley, in Sri Lanka and in some parts of Bangladesh (Saini et al. 2007).
A long legged and long necked bird, comparable in size to a rangy domestic turkey.
The iridescent dark blue head, neck and breast of resplendent cock is familiar to all, offset by
a featherless white cheek patch and the fan shaped crest of wiry black quills topped by small
spatulate iridescent green tips. The mantle and back is brilliant golden bronzy green, each
feather margined with black, the lower belly is dull black, wing coverts barred black and grey
and the flight feathers are largely pale chestnut, unbarred. In light the under-wing coverts are
black. The tail is dully grey brown and wedge completely hidden under the greatly elongated
and decorative upper tail coverts which form its well-known train (Jain and Rana 2013). The
Introduction
5
train consist of bronze green feathers, nearly all ending in the ocelli which are the famous
peacocks eyes each one roughly pear shaped and broadest at its base comprising an inner
purplish, black indented pupil surrounded by azure blue and the whole framed by a coppery
disc narrowly edged, again with an outer rim of green and dark bronze. The long powerful
thighs are buff and the legs and feet are horny brown. Female have a crest similar to the male
but the crown and nape is rufous brown and the rest of the neck and back grey brown with
metallic green gloss to the lower neck. The belly is dull white; flight feathers are chestnut and
the tail dark brown with whitish tips. There is some scale pattern to the flank feathers which
have glossy green tips. In both sexes the iris is dark brown and the actual white cheek patch
consist of bare skin. Males have a strong spur on the tarsus (Ramesh and McGowan 2009).
Global biodiversity of birds has declined markedly over the past 40 years (Hanotte et
al. 1991). The population of vertebrates in tropical regions specially in southeast Asia are
considered to be decreased tremendously which may result in extinction of many species till
next 50 years (Freeman and Hare 2015). There are two main reasons suggested behind the
decline in number of vertebrates in southeast Asia (Jain and Rana 2013). Continuous
deforestation may result in permanent loss of natural habitats of vertebrates, is the major
reason of declining the population of vertebrates in southeast Asia (Zhou et al. 2015). While,
uncontrolled preying and hunting of vertebrates for food, medicines and trades also resulting
the decrease in population of vertebrates (Weiss et al. 2011).
Very few studies have suggested the actual cause of decline in population of
vertebrates, still a lot of work has to be performed to quantify the causes of declining the
population of vertebrates (Harihar and Fernandes 2011). Weiss et al. 2011 has suggested that
development of policies on national level to conserve the natural habitats of animals may
result in meaningful increase in number of animals. Hence, adaptation of policies on national
and international level to increase the natural habitats of animals and vertebrates may result in
Introduction
6
their conservation and protection. Moreover, uncontrolled hunting and preying must be
discouraged (Takahashi and Hasegawa 2008; Munir et al. 2012).
Biological characteristics are play a key role to identify the species which is the most
important thing to conserve the animals but it cannot be achieved without morphologically
cryptic species are accurately identified. Previous studies in molecular genetics abled us to
investigate the relationship between cryptic and identification and their structural features to
differentiate among them (Murari et al. 2005). Although, behavioral study of animals in the
wild is a challenging job because we can observe the animals closely in wild condition.
However, the time span devoted to maintenance is a strong genetic basis, in captive condition
researchers can observe the behavior of the birds or animals more easily as compared to wild
(Teixeira et al. 2012). In Indian peafowl, sexual dimorphism is a definite prerequisite for
behavioral part of selection and directional selection, which selects evolution of such
distinguishing characters as the peacock‟s tail. The evolution of sexual dimorphism applied to
the forces of sexual selection acting upon both sexes of species. In male and female Indian
peafowl, dimorphic plumage signals and the presence of crest are related to sex (Sahajpal and
Goyal 2008). Normally, males are virtually suppliers of unlimited, extremely small and
inexpensive gametes in the reproductive process. In contrast, females produce large, nutritive
and expensive gametes in far lower quantities, a quality that actually dictates their role as the
limiting reproductive reagent. The female of polygamous pavo normally lay clutches of six
and often more than six eggs (Pabisch et al. 2010). In addition, females provide the possible
parental care to their offspring, hence reproductive success is dependent on the basic factors
like number of eggs produced and how they nurturing their young ones towards maturity
(Hart, 2002).
The tail ornamentation is the most interesting factor among peacocks. It can merely be
considered as a beneficial trait in intra-sexual selection but actually it is a poor weapon in
Introduction
7
maneuvering sexual copulate successfully (Hanotte et al. 1991). Due to this reason, females
have over reproduction; peahen preference is applying the basic driving force of sexual
selection over peacocks (Miller et al. 1998). Feeding is an important biological characteristic
as it is observed that supplementary feeding in hunting grounds may affect the number of
certain bird species to a greater extent (Khulape et al. 2014). As a result, pheasants in
captivity provided with extra feed are bred and reared under control and are released into the
wild after a specific time span, similar to broilers (Harikrishnan et al. 2010). Specific
deficiencies in proteins and in certain nutrients resulted in retard growth, poor feathering and
less resistant to drastic weather (Hanotte et al. 1991; Gupta et al. 2005; Ramesh and
McGowan 2009; Khulape et al. 2014).
5
CHAPTER 2
REVIEW OF LITERATURE
2.1 Indian Peafowl (Pavo cristatus)
The Indian peafowl (Pavo cristatus and Pavo muticus) natively distributed in Asia except
Congo Indian peafowl (Afropavo congensis). Congo Indian peafowl (Afropavo congensis) is
endemic to the Democratic Republic of Congo in central Africa. The Indian peafowl has
three bird species in the genera Pavo and Afropavo of the pheasant family, Phasianidae
(Araki et al. 1989).
2.2 Introduction
Across the world, the blue or Indian peacock is recognized and regarded as a most
glamorous bird. Remarkably, it is not generally known that there is an even more splendid
green peacock and also a much smaller, dumpier, African peacock that does not have the
spectacular train. The blue Indian and green peacocks are two of the most beautiful birds in
the world (Munir et al. 2012). The blue peacock was so much admired that it was taken from
its native haunts in India and Sri Lanka, thousands of years ago and gradually distributed
around the western world. We do not know when it was first introduced into Britain, but it
was a long time ago. That such an exotic bird became so familiar as an inhabitant of our
gardens and barnyards is a tribute to its hardiness and adaptability as much as to its beauty
(Ramesh and McGowan 2009).
Apart from the wonderful brilliant plumage and extraordinary performance in raising
its long train in a remarkable display, this is a bird of distinctive character. We think of the
peacock as a thoroughly masculine bird, proud, showing off, flamboyant and aggressive
(Parasharya and Mukherjee 1999). We accuse him of strutting proudly, even referring to him
when displaying as being „in his pride‟. His propensity to show off has recently been evoked
to describe the manner in which human youths behave. It has been found that separating boys
Review of Literature
6
from girls into different classes at school has improved the academic results of both.
Educationalists put this down to removing the „peacock effect‟ (Nasser et al. 2015).
The most great normal for the blue and green peacocks is their capacity to lift their
prepare plumes into an immense circular segment, 1.8–2.1 meters wide, and stroll round with
this show, shining and rattling the quills. It is a stunning presentation for the human watcher,
in spite of the fact that the peahen has all the earmarks of being aloof (Johnsingh and Murali
1980). She carries on as if she has seen it ordinarily some time recently, day after day, week
after week, every springtime – as indeed she has. Her only apparent response is to peck
studiously at minute particles of food on the ground. This studied indifference has intrigued
scientific observers, including Charles Darwin (Mushtaq-ul-Hassan et al. 2012).
In the 1990s scientists made careful observations of females and their choice of mates
and proved that the apparently disinterested and otherwise engaged females are actually far
more observant than would appear (Ismail et al. 2010). They choose for mates, and lay more
eggs for, males with the largest trains. It is three years before a peacock‟s train attains its full
length, largest eyespots (ocelli) and most intense color. The peahen‟s discerning eye carefully
selects the most glamorous and mature male, rejecting younger, less splendid birds
(Takahashi and Hasegawa 2008).
The peacock is perceived as an image in the religions of its characteristic Asian
natural surroundings, where it is a vehicle for divine beings in the Hindu and Buddhist
religions. It is additionally the subject of pixie stories, myths and legends (Trivedi and
Johnsingh 1996). In the west, a standout amongst the most acclaimed legends about the
peacock, known to both Greeks and Romans, is how it acquired the eye-spots in its train
feathers. The chief god Jupiter and his wife the goddess Juno were responsible for the
remarkable ocelli being placed in the peacock‟s plumage. Jupiter could never resist a
beautiful human maiden, and this made Juno extremely angry when she caught him out
Review of Literature
7
(Yorzinski et al. 2015). One of these unfortunate females was Io, whom Jupiter had to
transform quickly into a cow in an attempt to hide her from his wife‟s anger. Juno demanded
the cow be given to her and charged her cowherd, Argus, who had one hundred eyes all over
his body and so never slept, with the task of guarding Io. Argus was killed and Juno
preserved his hundred eyes by transferring them onto the train feathers of her royal bird, the
peacock (Zhou et al. 2015).
Other stories about the peacock‟s train, its voice and even its feet abound in the
literature of both east and west, but awareness of the presence of the peacock in our midst has
varied in intensity over the centuries. There have been several periods when the peacock has
been prominent (Ashok and Goyal 2013). It was present in early Christian art in illustrated
manuscripts and paintings of biblical scenes such as the Garden of Eden, the Creation and
Noah‟s Ark. In medieval manuscripts the illuminators reveled in the opportunity to paint all
the bird‟s intense colors. During the Middle Ages, it was a source of feathers for fletching
arrows and for personal adornment of men‟s helmets and hats, and on heraldic devices
(Somes Jr and Burger 1993).
During the seventeenth-century Golden Age of Dutch and Flemish painting, the
peacock was frequently present in pictures of both the barnyard and aristocratic gardens.
Little was seen of peacocks for the next century, and then they became important again for
embellishing millinery this time for women (Yasmin, 1997). The excessive use of their
feathers for this purpose contributed to the establishment of the Society for the Protection of
Birds in 1891 and the National Audubon Society of America in 1886. At the end of the
nineteenth century and the beginning of the twentieth they were such an integral and iconic
part of the Arts and Crafts and Art Nouveau movements‟ outpouring of metalwork, tiles,
glass, ceramics and other artifacts that it was impossible to avoid their image (Mushtaq-ul-
Hassan et al. 2012).
Review of Literature
8
For the last hundred years, they have again merged into the background, seen only in
large gardens where they can roam in a semi domesticated state. For thousands of years, these
magnificent birds have had a close association with man in diverse ways (Stewart et al.
1996). Few other bird species have had such a differed and broad history, holding an
unmistakable place in the way of life, craftsmanship and religion of eastern and western
countries. It has been a survivor, a truly remarkable bird reputed to have an angel‟s feathers,
the voice of the devil and the feet of a thief (Khulape et al. 2014).
2.3 Field Characteristics
Indian peafowl is a very familiar and almost universally known. The fan shaped crest of
spatula-tipped wire-like feathers together with the brilliant glistening blue neck and breast,
and the sweeping metallic bronze-green train, boldly ocellated with purplish black-centered
coppery discs or eyespots, make the cock unmistakable (Saini et al. 2007). Lower back light
bronze-green narrowly scalloped with black. Scapulars and outer surface of wings close-
barred with black and buff. A good deal of chestnut in wings (primaries and their coverts)
(Takahashi and Hasegawa 2008).
Female Indian peafowl are smaller, also peaked however without the general prepare.
Above head and scruff Rufus-dark colored. Rest of upper parts dark colored, faintly mottled
paler. Underneath, lower neck metallic green rather than blue; bosom bad tempered darker
sparkled with green; mid-region buffy white. Primaries dark colored with no chestnut as in
male. Juvenile male. Like grown-up female yet with the primaries to a great extent chestnut
(Nasser et al. 2015).
2.4 Biological Characteristics
Indian peafowl is inhabited in the dense forest of East Asia having two species, the
shared peacock (Pavo cristatus), and the Javanese peacock (P.musticus) peacocks. However,
the Congo peacock (Afropavo congensis), is found recently in Africa. Peacocks having an
Review of Literature
9
unusual trait are the Argus pheasant by the characteristic eye specks present on their minor
flight feathers, whereas, white peacock is a mutation of the common Indian peafowl. Whereas
using the word Indian peafowl, peacock indicate the male of a species and peahen indicate
the female (Ramesh and McGowan 2009).
The common or Indian Blue Indian peafowl has three varieties. The white Indian
peafowl having white feathers in its train along with ocelli barely visible. These are not
considered albinos as they are basically true breeders. In another variety known as pied
version, random white feathers are in the plumage due to the results of an incomplete
dominant gene (Ashok and Goyal 2013). Another variation results due to mutation with dark
feathers having blue and green tips, called the black-winged Indian peafowl. In addition,
Pavo cristatus is often hybridized with green Indian peafowl (Pavo muticus) so new mutation
in the plumage almost every year has been discovered in the last decade (Khulape et al.
2014).
At the start of the 20th century, green Indian peafowl (Pavo muticus) were widespread
and common across much of South-East Asia (Saini et al. 2007), so much so that they were
considered the second most abundant game-bird in Indochina after the red jungle fowl Gallus
gallus; (Ramesh and McGowan 2009). In the last few decades‟ green Indian peafowl numbers
have declined dramatically (Nasser et al. 2015). They are now considered to be extinct in
Malaysia (Somes Jr and Burger 1993) and restricted to isolated populations in India,
Thailand, Laos, China and Indonesia (Hanotte et al. 1991; Landman and Gruys 1998; Hart,
2002).
The green Indian peafowl (P.muticus) historically covers a large area in East and
Southeast Asia, from India to Indo-China and Java and may be extinct in some places such as
northeast India and Malaysia (Parasharya and Mukherjee 1999), although it is protected by
law in many countries like Burma, Indonesia, Malaysia and Thailand (Yasmin and Yahya
Review of Literature
10
1996; Yorzinski et al. 2015). Currently, the number of Indian peafowls is declining due to
excessive hunting for feathers, meat and chicks either for sale or for pet keeping. Indian
peafowl is considered as a vulnerable species in the revised IUCN red list. Although
peacocks never provide food for peahens during the courtship nor they offer post-mating
parental care, hypothesis are focused on the transfer of beneficial aspects to the peahen and
the advantage of „good parenting‟ are not sufficient to provide information about the
evolution of the male‟s tail (Trivedi and Johnsingh 1996). The complicating male liking
behavior in Indian peafowl can be considered as kin selection and cannot be considered as
direct selection (Landman and Gruys 1998; Parasharya and Mukherjee 1999).
Ornamentation in feather patterns is characteristic of Indian peafowl, high quality
ornament signals at high levels of resistance and immunity. The expression of ornaments may
be an indication of the ability of the bearer to its defense against disease causing pathogens
(Somes Jr and Burger 1993; Stewart et al. 1996; Takahashi and Hasegawa 2008). Food
quality is very necessary for the growth of the immune system during the first weeks of post
hatching period, because there is a quick development of leukocytes and seeding of lymphoid
structures during this period. During this period, any deficiency (or an excess) of nutrients at
this stage may abruptly affect the resistance of an individual in the later life (Yasmin, 1997).
It is estimated officially that above 1/3 of pheasant species are itemized as in risk of
elimination. Pheasants are mostly a source of refreshment and desirability for mankind. The
reason behind this interest is their charming sound and beautiful feathers. This beauty of
nature is easy to trap or shoot and obviously, their meat is a rich source of protein (Freeman
and Hare 2015).
Indian peafowls are the source of cultural societies, skill, conviction, and folklore of
dissimilar ethnic crowds in Asia (Stewart et al. 1996). Indian peafowl is a potential source for
improving ecotourism, hunting, and public betterment of native societies in emerging
Review of Literature
11
republics in their natural habitats and in captivity as well (Mushtaq-ul-Hassan et al. 2012).
The Common Indian peafowl is under threat due to heavy demand for feathers and wild meat,
during cropping season a source of conflict, advancement in green revolution such as
chemical fertilizers and pesticides and habitat destructions. Approximately an adult peacock
has shed their feathers from August to onward, fully developed new tail feathers appear in
February (Samour et al. 2010).
The peacock exists very precariously in the extreme north eastern border regions of
the Punjab as well as the south-eastern corner of Sindh. In the 1940s and up to independence
it was fairly widespread in the better water parts of Sindh (Nasser et al. 2015) such as around
Mirpurkhas, Khipro, the Makhi Dhand in Sanghar district, and the Thar desert. It was also
quite common in Hyderabad district. K. Eates consider that this whole population was feral
and that it had been introduced by the Mirs of Talpur after the Kalhora dynasty was
overthrown. After independence, it was rapidly exterminated by local hunters who had no
religious compunctions about shooting a bird, regarded regarded as sacred by the Hindu
villagers living in Tharparkar. It survives today still in good numbers in the extreme southern
border regions of tharparkar around Islamkot and Nagar Parkar where predominantly Hindu
communities still afford it protection (Ramesh and McGowan 2009). Since this population
extends widely throughout the Thar desert and into Rajasthan it seems probable, that it was
indigenous and not in fact introduced by the Talpur Mirs. Up in the Punjab there used to be a
small feral population around at Kallar Kahar in the Salt Range, given some protection
because of their habit of roosting by the local Mosque. In the foothills regions of Sialkot
district and further north in the Jhelum watershed, around Panjar and Kahuta a small
genuinely wild population survives very preciously (Munir et al. 2012).
2.5 Status, Distribution and Habitat
Review of Literature
12
Resident, locally Up to C. 1800 In. elevation (rarely even 2000 m.) in the outer Himalayas
and peninsular hills, practically throughout the subcontinent south and east of the Indus river,
including Jammu and southern Kashmir. Eastward to about Lakhimpur district in Assam (c.
95"E.), thence south through Nagaland, Manipur, Mizo (Lushai Hills), and Chittagong west
and south to the extreme tip of the Peninsula (Khulape et al. 2014). Also indigenous and
resident in Ceylon where locally plentiful in the low-country Dry zone. Introduced and now
fairly established with a semi-feral status in parts of Sind (W. Pakistan), e.g. Hyderabad,
Mirpurkhas, Umarkot and Sehwan areas. Very common and abundant in Gujarat and
Rajasthan and locally wherever protected on religious or sentimental grounds. Introduced
successfully also in the Port Blair area and elsewhere in the Andaman Is (Nakamura et al.
2009). In wild state affects moist- and dry-deciduous forest in the neighborhood of streams.
Where semi-feral, found in the precincts of villages and cultivation, in close association with
man (Samour et al. 2010).
2.6 General Habits
Keeps in small flocks or droves consisting usually of a cock plus 3 to 5 hens. Afterward the
breeding season segregated parties of adult males and of females with their immature broods
are more common. In the wild state the birds emerge cautiously into forest clearings and
ploughed or newly sown fields in the early mornings and late afternoons to scratch the
ground for food (Ramesh and McGowan 2009). An hour or so after sunrise they troop down
to the water to drink, tripping gingerly over the dry leaves, craning their necks at every step
and peering inquisitively through the bushes with the utmost circumspection. The birds also
drink at dusk before retiring to roost (Yasmin and Yahya 1996).
They spend the heat of the day in impenetrable thickets of lantana and the like.
Peafowl are possessed of phenomenally keen eyesight and hearing and are excessively wary
and wide. awake at all times, ducking their heads and slinking away through the undergrowth
Review of Literature
13
on the least suspicion (Takahashi and Hasegawa 2008). The cock's cumbrous-looking train,
carried horizontally a few inches above ground, is apparently no impediment to swift
progress through the thickest cover. The birds prefer to trust to their legs for escape, running
one behind the other when driven by beaters, and seldom take to wing except when flushed
by a dog or to cross some open river-bed or ravine (Ismail et al. 2010).
They rise with a loud flapping of wings, even an old cock with his long, heavy train
rocketing almost vertically to clear the tree-tops. When properly launched they fly fast and
strongly with rapid flaps and glides and dexterous twists and turns to avoid tree-trunks and
other obstacles (Araki et al. 1989). When surprised in the open with a brood of small chicks,
the hen pulls out the family to safety with amazing adroitness, crouching low with neck
outstretched to take full advantage of every little dip and unevenness in the ground for
concealment as she swiftly makes for cover in a manner inconceivable for so large a bird
(Ashok and Goyal 2013).
2.7 Roosting at Night
Peafowl roost at night in tall trees; the jungle resounds with their loud and ugly may-awe
calls at dusk and early dawn before the birds descend to reed. They often call from the roost
also during the night, and the crashing of a tree or a sudden clap of thunder is sure to release
an immediate burst of responsive caterwauling by all the cocks in the neighborhood (Dakin,
2011). Cocks are quick to detect the presence of the larger cats on the prowl and herald the
marauders' progress through the forest with loud warning alerts which are taken up by other
cocks and by langur monkeys (Black et al. 2010).
2.8 Food
Omnivorous. Seeds, grain, lentils, groundnuts, tender shoots of crops, flower-buds, berries
and drupes, wild fiq (Ficus glomerata and other species), centipedes, scorpions, lizards, small
snakes (once c. 20 cm. long), insects (e.g. locust hoppers, beetles, caterpillars), worms and
Review of Literature
14
grubs have all been more or less regularly found in crop and stomachs (Samour et al. 2010).
In and around villages also human excreta. In areas where it is semi-feral and abundant it is
destructive to cereal and groundnut crops in the highest degree, and a veritable scourge to the
small farmer, vying with the sacred monkeys in the work of devastation (Miller et al. 1998).
2.9 Voice and Calls
Normal ' crow ' of cock a loud and harsh metallic trumpet-like may-awe shrieked several
times, varying in pitch. Also a series of short, gasping screams ka-an . . . ka-an . . . ka-an (or
cain . . . cain . . . cain ponounced as in French) repeated 6 to 8 times rapidly with an inane
pumping action of the head and neck, well likened to ' something between the scream of a
hysterical old woman and the bray of a donkey (Bhattacharyya, 1971). Alarm calls when
suspicious and agitated a double-syllabled brassy kok-kok or cain-kok, and variations of the
same. This commonly uttered by a hen with neck-feathers fluffed out like a bottle brush and
repeated intermittently for many minutes, especially when leading a brood of chicks (Gupta
et al. 2005).
2.10 Breeding Behavior
Indian peafowl breeds from April through October. Indian peafowl is polygynous and
generally has two to three breeding peahens in its harem. A recent study of a feral population
suggests that Indian peafowl does not defend its harem due to small breeding territory. Clutch
size is normally 4 to 9 eggs in characteristic living space however in imprisonment the hen
lays 8-20 eggs and the brooding time frame is around 28-30 days (Landman and Gruys 1998;
Dakin, 2011; Dodia, 2011). Chasing of Indian peafowl is denied under the Punjab Wildlife
(Protection, Preservation, Conservation and Organization) Act-1974. The hostage
reproducing system of peacock was begun by the Punjab Wildlife Division a couple of
decades back. The main purpose of this program was to propagate the number of threatened
or endangered species in captivity and reintroduce them into their natural habitats. It also
Review of Literature
15
provided a research platform for researchers to boost understanding of the biology of this
species. These activities may provide important information for conservation efforts (Hart,
2002; Hollamby et al. 2003; Ismail et al. 2010).
Different experiments showed that nutrition has played a dynamic role in the early
development of the bird and supplementary diet is required for proper growth. Green feed
stuff and grains are introduced in order to mimic natural feeding conditions. As Indian
peafowl is omnivorous, so the protein content plays an important role in building different
biological characteristics e.g. growth rate of the bird (Harihar and Fernandes 2011). Most
evidences supported that improved growth of Indian peafowl offspring with elaborated trains
is related to higher amount of fat reserves in peacocks with longer trains, and fewer
heterophils in peacocks having long trains and frequent displays. Thus, the elaborate trains
and display of the peacock act as an honest signal of good genes and male quality at the time
of mating (Miller et al. 1998; Parasharya and Mukherjee 1999; Nasser et al. 2015).
Munir et al, (2012) kept the free-range population at Khanewal in citrus orchard, from
May to August. The males each had a particular territory or dancing ground where they
would fan their traits in the early morning and late afternoons, quivering their erected upper
tail over their backs with an audible rustle of the quills and lifting their feet up and down in a
sort of dance. One individual repeated displayed towards a pet rabbit, ignoring female
peacocks feeding nearby. Females laid under a pile of brush wood e.g. cotton sticks with no
lining to the hollow scraped out of the bare earth. The eggs are creamy buff, unmarked and
the usual clutch is 4 to 6 eggs (Nakamura et al. 2009; Mushtaq-ul-Hassan et al. 2012).
During the courtship behavior, the crested male of peacock shows his lengthened
higher tail coverts a wonderful green and gold erectile train decorated by blue-green “eyes”
earlier the dull-plumaged peahen. The declaration of male sexual decorations is influenced by
nourishment; the trimming is condition-subordinate and may hail both phenotypic and
Review of Literature
16
genotypic quality (Khan et al. 2009; Ismail et al. 2010; Khulape et al. 2014). The splendid
yellow, orange and red hues generally found in the brushes, wattles, mouths, skin fixes, eyes
and plumes in fowls are as a rule brought about via carotenoid pigmentation. In a few winged
creature animal categories these carotenoids based hues work as decorations and are utilized
by females for choosing mates. However, during” train rattling” display, a basic pre-
copulatory action, male is directed 45˚ right to the sun in front of female to enhance the
eyespots on feathers (Landman and Gruys 1998; Nakamura et al. 2009).
There is a positive correlation between an Indian peafowl‟s train and success of
Indian peafowl mating. This correlation is actually due to female‟s preference for well
elaborate trains on their mates (Dodia, 2011). Expenditure of a lot of energy by male to
produce and maintain elaborated good train conditions, will not only result in the form of a
trade-off between a longer train but also avoid predators and search for food. Mating success
was normally more successful with the large number of eyespots also called ocelli on the
train of the peacock (Parasharya and Mukherjee 1999; Jaiswal et al. 2013).
The peacock‟s tail is long-standing examples of sexual selection as long tail is an
exaggerated character that hinders the individual‟s survival but no doubt it is of increased
reproductive success. Researchers agreed that the peacock attract females for mating by using
its ornamental tail (Crawshaw and Boycott 1982; Chumbe et al. 2015). The ornamentation of
the peacock‟s train has been measured with the number of eyespots, train length and area of
eyespots, density of eyespots, proportion of feathers with eyespots, diameter of eyespots, and
eyespot color and iridescence. Currently, the consensus is that the peacock‟s train is a
handicap but it is also considered an indicator of good genes (Araki et al. 1989; Ashok and
Goyal 2013; Zhou et al. 2015).
Review of Literature
17
2.11 Color of Bare Parts
Iris dark hazel brown. Bare facial skin livid white Bill dark horny, darkest along culmen and
at tip. Legs and feet greyish brown to dark horny brown; claws blackish. While chicks are
Pale buff; a dark brown mark across the nape from one eye to the other; back darker rufous-
brown; wing pale dull chestnut mottled with brown (Barbieri et al. 2012).
2.12 Miscellaneous
The peacock is known sacred in Hindu tradition as the transport of the god Kartikeya the son
of Lord Shiva and his partner Parvati, and a brother of elephant-headed Ganesha. It is,
besides, a subject of voluminous folklore throughout the country. For an account of its place
in mythology and Sanskrit literature and poetry (El-Shahawy, 2010). The long ocellated
feathers of the peacock's train are shed after the breeding season and picked up and collected
in large quantities by villagers for export chiefly to Europe and America, and for being made
up locally into fans and other gaudy gimcracks (Hart, 2002). The export has now been
prohibited as a measure of protection for the species, a step perhaps unnecessary considering
that the trade was almost entirely confined to dropped feathers available in marketable
quantities only in areas where the birds are already well protected by sentiment and therefore
abundant enough to make the collection of the feathers economically worthwhile. An average
adult cock's full train contains about 200 + feathers. In 1963 the peacock was publically titled
as the National Bird of India (Harihar and Fernandes 2011; Jain and Rana 2013).
2.13 Genomics and Phylogenetic Analysis
Progresses in sub-nuclear methods, especially the improvement of the polymerase chain
response, have made investigations of vertebrate genomes progressively common sense, and
have deterred the necessity for completely prepared sub-nuclear science research centres.
Among ornithologists, this has brought about a touchy increment in investigations of
systematics and populace hereditary qualities (Amoudi, 1988).
Review of Literature
18
It is additionally underscored that the avian genome holds homologs to mitochondrial groups
in at any rate a few animal types, and this may have genuine ramifications for the
understanding of mtDNA informational indexes utilized as a part of populace hereditary and
precise reviews. In the meantime, such events show significant new chances to pick up a
superior comprehension of grouping advancement in both mitochondrial and nuclear
genomes (Hollamby et al. 2003).
2.14 Mitochondria: An Ancient Legacy
In 1922, it was suggested that mitochondria rose by endosymbiosis, a suspected that was
defended and created by (Ansari, 1957). They suggested that mitochondria began when a
protoeukaryotic cell inundated or was entered through a high-affect bacterium. The
endosymbiont hypothesis was at initially solved with much doubt, be that as it may it has in
this way got strong support and is by and by comprehensively recognized, with common
contention now focusing on whether mitochondria are monophyletic or polyphyletic in cause
and on the character of the main symbionts (Pabisch et al. 2010; Kumar et al. 2013). The
acquirement of such a high-affect endosymbiont is among the most basic events ever, for the
relatives of those affiliations now incorporate all eukaryotic life, both single-celled and
multicellular (Tripathy et al. 1972).
Among different qualities that are definite of this formative legacy is the way that
mitochondria pass on their own (for the most part circuitous) self-rulingly impersonating
genomes. The small size of this genome in animals in regard to that of surviving minute
living beings (around 1% as enormous) is presumably the eventual outcome of transfer of
characteristics that were abundance with those in the atomic (host) genome, and of the
coincidental trade of down to earth qualities from the mitochondrial genome to the atomic
genome in our endo-profitable ancestors (Krautwald and Schildger 1986; Jaiswal et al. 2013).
The exposure of a round genome inside the mitochondria of most eukaryotes upheld the
Review of Literature
19
endosymbiont hypothesis, and additionally it perceived a little and viably detached wellspring
of innate information outside of the far reaching and complex atomic compartment (Weiss
and Kirchner 2010).
2.15 The Vertebrate Mitochondrial Genome
Since the change of recombinant DNA frameworks, formative researchers have made the
vertebrate mitochondrial genome a champion among the most generally considered on the
planet. It is obvious that a principle thought in the fundamental assurance of mtDNA for
formative audits was the relative straightforwardness with which it could be purified and
controlled in the exploration office, keeping to its high copy amount and supercoiled
adjustment, which licenses segment from direct (atomic) DNA. (Krautwald and Schildger
1986) assessed that there are 800 mitochondrial genomes for every cell, and an ordinary of
2.6 genomes for each mitochondrial organelle, inside refined human lung fibroblast cells, Liu
et al. 2005 evaluated 2600 genome copies for every cell in fundamental bull like tissue
culture cells. Gages for other tissue sorts change comprehensively.
Once purified, mtDNA course of action complexities between species or individuals can be
assembled by suggestion using repression endonucleases to make discrete pieces that can
then be contemplated by methods for electrophoresis through agarose gels, or clearly by
DNA sequencing (Murari et al. 2005). The polymerase chain reaction (PCR) has dynamically
supplanted such cleaning methodologies by permitting the prompt upgrade and sequencing of
mtDNA from unpurified sources. Regardless, as atomic copies of mitochondrial qualities
have been noted in winged creatures, usage of straight cleansing methodologies will be of
ongoing with regard (Weiss et al. 2011).
As a bit of the essential transformative audits using refined mtDNA were done, it wound up
obviously apparent that, other than its straightforwardness of separation and little genome
measure, there were various distinctive central focuses to think the mitochondrial genome
Review of Literature
20
(Sahajpal and Goyal 2008). At first, it is maternally gained. Second, there is no prompt
confirmation that it can recombine with other mtDNA particles. This suggests vertebrate
mtDNA is gone ahead through female family histories in a clonal way with no level
"mixing," and this makes it more straightforward to duplicate a transformative history of this
molecule than for the atomic genome (Vijayarani et al. 2010). While Hanotte et al. 1991 have
recognized low levels of paternal "spillage" of mtDNA between two sorts of mouse, paying
little respect to whether such a finding can be extrapolated to intraspecific spillage is up 'til
now not clear. (Chumbe et al. 2015) exhibited that biparental legacy occurred in interspecific
Drosophila crosses, however did not occur when intraspecific crosses were done. Third, at the
progression level, mtDNA has been seemed to grow quickly in regard to DNA in the atomic
genome. This is an advantage for masses contemplates, however over the long haul
transforms into a hazard as the significance of phylogenetic relationship fabricates (Teixeira
et al. 2012). The key mitochondrial genomes to be sequenced totally join human, mouse,
dairy creatures and frog (Murari et al. 2005; Pabisch et al. 2010). Not only was the quality
substance saw to be indistinct, yet the quality demand was the similar, and this, in
combination with a comparative finding in hard fish, provoked the supposition that all
vertebrate mtDNA genomes were vague in both respects (Sahajpal and Goyal 2008). As more
conspicuous amounts of genomes have been sequenced, clearly quality demand, especially of
tRNA qualities, is not reliable. For example, among tetrapods reports of tRNA exchange have
been prepared for marsupials, frogs, and crocodiles (Weiss et al. 2011; Zhou et al. 2015). Lee
and Kocher (1995) have exhibited that in an outgroup to the Osteichthyes, the sea lamprey,
there have been a couple fluctuations in quality demand near the putative control locale, some
of which have included tRNAs. All vertebrate mtDNAs contain 22 tRNAs, 13 protein-
coding regions, 2 rRNAs, and 1 or 2 (lamprey) far reaching noncoding sections. The genome
is sorted out capably. Introns are missing, and intergenic spacers are pretty much nothing,
Review of Literature
21
normally under 10 bp (Chumbe et al. 2015). Once in a while, qualities cover in different
examining plots, and there has even been a diminishing in the measure of some stop codons
to perhaps two or three bases (T or TA). Such codons are post transcriptionally done with the
extension of a 3' poly (A) progression (Murari et al. 2005; Teixeira et al. 2012).These traits of
vertebrate and various nonvertebrate mitochondrial genomes have provoked the proposal that
intermolecular assurance for conservativeness could be the "primary purpose" that results in
such a calm quality game-plan, expecting that tinier molecules copy more rapidly than greater
ones, all else being identical (Rao and Acharjyo 1979; Sahajpal and Goyal 2008; Pabisch et
al. 2010). Demonstrate in support of this starts from recognitions that human mitochondrial
genomes passing on eradications can, between times, augment in repeat concerning fully
grown genomes both in vitro and in vivo (Samour et al. 2010; Teixeira et al. 2012).
Review of Literature
22
Statement of Problem:
Population of Indian Peafowl is declining. Limited research has been conducted to identify
the genetic variations in different varieties of peafowl. Breeding behavior and phylogenetic
studies will help us to conserve the species. The present study is therefore planned to study
the breeding behavior and genetic diversity of different varieties of Indian peafowl present in
Pakistan reared in captivity. The current research will focus:
To find out genetic variations of Indian peafowl.
To study the breeding behavior of different varieties of Indian peafowl.
To develop the evolutionary relationships between different Indian peafowl variants.
To construct a phyletic lineage between different varieties of Indian peafowl.
23
CHAPTER 3
MATERIALS AND METHODS
The present study was planned to investigate breeding behavior and genetic diversity of
various kinds of Indian peafowls reared in captivity in central Punjab during the period of one
year. Indian peafowl from five government zoological gardens and five private sectors was
studied during research. Breeding behavior of Indian peafowl was studied by observing
different morphological characteristics which play a great role in the breeding of Indian
peafowl. Female preference is based on visual, auditory and olfactory for certain male
characters. As breeding season is geographically variable so in order to know about breeding
behavior, site and structure of the nest, clutch and incubation period, season of breeding and
temperature was monitored under captive conditions. Male courtship display was monitored
during the time period of study. Peacock tails are energetically costly to produce and
maintain but accurately signal the immune system strength of the males to potential mates
(Liu et al. 2005). Peacocks perform courtship ritual which is a sum of behaviors involving
calls and the presentation of the visual train ornament in the presence of females. Display
bouts in lek sites was observed during study along with recording of specific time period
(Dakin, 2011).
Breeding Behavior
To record the breeding behavior of Indian peafowl, birds were observed for one year,
especially during their mating season, during when birds starts to display in the morning until
the mid-day, when they stop to display. Usually time period of our observation was (7:00 am
to 4:00 pm). Data such as; when a female approach a male and the male started to display and
begin to shiver his train at the female, following copulating behavior of both male and female
were recoded. After copulation, behavior of male and female Indian peafowl was also
recorded. Any aggressive contact from any side also checked.
Materials and Methods
24
Common Behavior
We investigated the common behavior of Indian peafowl such as feed, stand, walk, run, sit,
display, preen, flight, fly and call. From scan records, proportion of time allocated for each
type of behavior of each bird from both sector was calculated for a period of six weeks.
Genetic Diversity
The genetic diversity of Indian peafowl has never been studied earlier. This study will help us
to investigate the diversity of allelic frequencies of different microsatellite DNA markers or
mitochondrial DNA (mtDNA) markers. After the study of genetic diversity, we will be
enable to conserve the allelic frequencies in a population rather than the population number.
For this purpose, genomic (gDNA) will be extracted from blood.
Genomic (gDNA) Extraction
For extraction of DNA, 5ml blood was collected in EDTA containing vacutainer from wing
vein of each Indian Peafowl (1 sample from each: five government zoological gardens and
five private sectors). DNA was extracted using the Nucleon BACC2 kit (GE Health Care-
Amersham). The components of the kit are as follow:
Table 3.1: Components of the illustraTM
Nucleon BACC2 RPN850)
Sr. No. Reagent Quantity
1. Reagent A 420 ml
2. Reagent B 110 ml
3. 5 M Sodium Perchlorate 26 ml
4. Nucleon Resin 16 ml
Protocol
1. 50μl blood was added in a tube having EDTA.
2. Using aseptic procedure, 4times the volume of Reagent A to the blood sample, was added
in tube.
Materials and Methods
25
3. Rotary mixed the sample for four minutes at room temperature.
4. After this, centrifuged the sample at 1300 g for 5 minutes.
5. Supernatant was discarded.
6. Pellet was formed.
7. 350μl of reagent A was added to the pellet.
8. Vortexed briefly to re-suspend the pellet.
9. Suspension was transferred to a 1.5 ml micro-centrifuge tube.
10. 100 μl of sodium perchlorate solution was added in tube for Deproteinisation.
11. Mixed solutions by hand.
12. Capped tube was inverted for at least 7 times.
13. 600 μl of chloroform was added to above mixer for DNA extraction.
14. Mixed the mixture by hand.
15. Capped tube was inverted for at least 7 times.
16. Without remixing the phases, 150 μl of Nucleon resin was added to above mixture.
17. Centrifuged it at 350 g for 1 minute.
18. Without disturbing the Nucleon resin layer (brown in color), the upper phase
(approximately 450 μl) was shifted to a 1.5 ml microcentrifuge tube.
19. 2 volumes (900 μl) of cold absolute ethanol was admixed by inversion until the
precipitate appears (gDNA precipitation).
20. For gDNA washing, centrifuged the precipitated gDNA, at 4000 g for 5 minutes.
21. Supernatant was discarded.
22. 1 ml cold 70% (v/v) ethanol was added and mixed several times by inversion.
23. Re-centrifuged and discarded the supernatant.
24. Pellet was air dried for 10minutes, to remove ethanol.
25. DNA was re-dissolved in TE buffer 50μl.
Materials and Methods
26
26. The tubes were placed in 37°C shaker incubator overnight to dissolve the DNA. The
Parafilm strips were wrapped around the caps of the tubes.
27. Temperature of shaking water was set to 70°C, and tubes were placed in it for 1 hour to
inactivate nucleases.
28. After cooling to room temperature, tubes were spinned for very short duration.
29. Side and cap of 1.5 ml eppendorff tubes were labbeled. gDNA aliquots were taken in
duplicate and stored at -20°C according to number in marked cryoboxes.
30. Quantification of extracted gDNA was done by using Nanodrop 2000c
spectrophotometer.
Quantification of Extracted DNA
To assay the gDNA yield and to check gDNA purity, Nanodrop (Thermo Scientific
NanoDrop™ 2000/2000c Spectrophotometer) was used. Absorbance ratios at 260 nm/280 nm
were measured, find out the purity of the extracted gDNA.
gDNA amount in dilution was calculated as follow:
Amount of gDNA (ng/μl) = Absorption at 260 x 50 x DF
DF= Total volume of dilution / Volume of stock gDNA solution in the dilution
From the calculated amount of gDNA in Nanodrop spectrophotometery, 5ng/μl dilution will
be prepared in 100 μl volume.
The dilution was prepared as follows:
V1= (C2×V2)/C1
Where,
C1 = Amount of gDNA in stock solution.
V1 = Volume of gDNA of stock solution to be diluted.
C2 = Amount of gDNA in dilution to be prepared (5 ng/μl).
V2 = Volume of gDNA dilution to be prepared (100 ml).
Materials and Methods
27
All the dilutions were stored at 4°C.
Amplification of gDNA Samples
For amplification of cytochrome b gene with universal primers (Verma and Singh 2003)
(Table 3.1), extracted gDNA was used as template. A total of 50μl reaction mixture (Table
3.2) was prepared by using 5.0μl DNA, 5.0 μl PCR buffer (10×), 5 μl dNTPs (10 mM), 2 μl
of each primer (10 pmol/μl), 0.5 μl Taq DNA polymerase (5U/μl) and 30.5μl of Nuclease free
water (DEPC, Invitrogen, USA).
Table 3.2: Universal Primer sequences for cytochrome b amplification
Primer Sequence (5’---------3’)
mcb398 TACCATGAGGACAAATATCATTCTG
mcb869 CCTCCTAGTTTGTTAGGGATTGATCG
Table 3.3: PCR reaction mixture for cytochrome b amplification
Reagents Volume (μl)
Template 5.0
Primer (mcb398) 2.0
Primer (mcb869) 2.0
10x PCR buffer 5.0
10 mM dNTPs 5.0
Taq DNA polymerase (5U/μl) 0.5
DEPC 30.5
Total volume 50.0
The amplification of gene was carried out according to the following program, initial
denaturation at 95 ºC for 10 minutes followed by 35 cycles of each denaturation at 94 ºC for
45 sec., annealing done at 54 ºC for 1min, first extension was done at 72ºC for 2 minutes and
last extension at 72 ºC for 10 minutes (Gupta et al. 2005) (Table 3.3).
Materials and Methods
28
Table 3.4: Thermo cycler conditions for cytochrome b amplification
Stages
PCR Conditions
Cycles
Temperature Time
Initial Denaturation 95 ºC 10 min 1
Denaturation 94 ºC 45 sec
35 Annealing 54 ºC 1 min
Extension 72 ºC 2 min
Final Extension 72 ºC 10 min 1
Sequencing and Analysis
To identify the genetic diversity between the Pavo cristatus available at different sites, the
cytochrome b gene sequences were performed. The obtained sequence results were utilized
for the homology analysis through NCBI-BLAST and the comparative analysis of the
sequence with the already published sequences available in NCBI Data Bank was done.
Statistical analysis
The data taken was analyzed by using Repeated Measure Design ANOVA to compare
different groups with the help of Statistical Package of Social Sciences (SPSS 17.0, Chicago,
IL). The pattern of data will be demonstrated by using graphical presentation. Differences
was considered significant at P < 0.05.
29
CHAPTER 4
RESULTS
4. Breeding Behaviors
In this study, we observed 5 pairs of Pavo cristatus at each Government zoological gardens
and at private sectors. We observed the frequency of mating, average eggs per season,
breeding season, range time to hatching, breeding interval, and average age at sexual or
reproductive maturity of both male and female, and average time to independence, and we
got following results.
4.1 Frequency of Mating of Female Pavo cristatus
Frequency of mating of female Pavo cristatus described in following table. Data showed that
number of female copulated only once at government and private sector were 17 and 19
respectively with 68.00 % and 76.00 % proportions respectively at each sector. While,
number of female birds copulated more than one time at both government and private sector
were 8 and 6 respectively with 32.00% and 24.00% proportion at each sector respectively.
Data represent that although there was no significant difference (p>0.05) at both government
and private sector regarding the frequency of mating of female Pavo cristatus
Table 4.1: Frequency of Mating of Female Pavo cristatus
Parameters
Sector
P-Value Government
Zoological
Gardens
(n= 25pairs)
Private Sectors
(n= 25pairs)
Number of female copulated only
once 17 (68.00 %) 19 (76.00 %)
0.5287 Number of female copulated more
than one time 8 (32.00 %) 6 (24.00 %)
Results
30
Figure 4.1: Graphical representation of frequency of mating of female Pavo cristatus
Results
31
4.2 Number of Copulation of Female Pavo cristatus
Table 4.2 showed that at government zoological gardens, out of total 8 female Pavo cristatus
who copulated more than one time, three female copulated two times, one female copulated
three times, two female copulated four times, one female copulated five times and one
copulated nine times. Similarly, at private sectors out of total 6 female Pavo cristatus who
copulated more than one time, two female birds copulated two times, two female birds
copulated three times, and one female bird copulated six times while one copulated eight
times.
Table 4.2: Number of Time Copulation of Female Pavo cristatus
Sector Number of Female
Number of Times
Copulation
Government Zoological
Gardens
3
1
2
1
1
2
3
4
5
9
Private Sectors
2
2
1
1
2
3
6
8
Results
32
4.3 Number of Mates of Female Pavo cristatus
Table 4.3 showed that at government zoological gardens, out of total 8 female Pavo cristatus
who copulated more than one time, five female copulated with two male birds, two females
copulated with three male birds while only one female copulated with five male birds.
Similarly, at private sectors out of total 6 female Pavo cristatus who copulated more than one
time, three female birds copulated with male birds, two female birds copulated with three
male birds while only female copulated with six male birds.
Table 4.3: Number of Mates of Female Pavo cristatus
Sector Number of Female Number of Mates
Government Zoological
Gardens
5
2
1
2
3
5
Private Sectors 3
2
1
2
3
6
Results
33
4.4 Common Behavior
We investigated the common behavior of Indian peafowl such as feed, stand, walk, run, sit,
display, preen, and call. From scan records, proportion of time allocated for each type of
behavior of each bird from both sector was calculated for a period of six weeks. We found
following results. Data showed that a significant difference (p<0.05) was found between male
and female Indian pea fowl regarding the feeding, standing, walking, running, sitting, preen
and call behavior at both government zoological gardens and private sectors.
Table 4.4: Percentage of time (hours) allocated for each behavioral activity of Indian
peafowl for a period of six weeks (breeding season)
Behaviour Sector
Government Zoological Gardens Private Sectors
Male Female P-value Male Female P-value
Feeding 24.97 54.56 < 0.0001 25.18 54.44 < 0.0001
Standing 36.91 19.93 < 0.0001 37.03 20.21 < 0.0001
Walking 12.97 20.50 < 0.0001 13.07 20.58 < 0.0001
Running 0.20 0.75 < 0.0001 0.19 0.76 < 0.0001
Sitting 0.20 0.17 < 0.0001 0.20 0.18 < 0.0001
Display 15.76 --- --- 16.02 --- ---
Preen 2.05 1.79 < 0.0001 2.13 1.84 < 0.0001
Call 2.93 0.06 < 0.0001 2.97 0.08 < 0.0001
Results
34
4.5 Average Eggs Per Season
Average clutch size of Pavo cristatus at government zoological gardens was 8.6 ± 0.55 and at
Private Sectors was 7.24 ± 0.61. It showed that there was non-significant difference (p>0.05)
between the Indian Peafowl available at different location.
Table 4.5: Average Number of Eggs of Pavo cristatus
Sector No. of Eggs (Mean ± S.E.) p-value (p<0.05)
Government Zoological Gardens 8.6 ± 0.55
0.096
Private Sectors 7.24 ± 0.61
S.E.: Standard Error of Mean
Results
35
Figure 4.2: Average clutch size of Pavo cristatus at both government zoological gardens
and private sectors
Results
36
4.6 Breeding Season
As we know that, Indian Peafowl breed from April to September, and following results are
according to the data published. We noted that most of birds at government zoological
gardens breed in month of May, while most of birds at private sectors breed in the month of
August.
Table 4.6: Breeding Season of Pavo cristatus
Month
No. of birds breed at
Government Zoological
Gardens
No. of birds breed at
Private Sector
Total
April 4 2 6
May 9 2 11
June 5 4 9
July 3 6 9
August 3 8 11
September 1 3 4
Total 25 25 50
Results
37
Figure 4.3: Breeding behavior of Pavo cristatus in different months
Results
38
4.7 Range Time to Hatching
The mean range time to hatching of Pavo cristatus at government zoological garden and at
private sector was 27.92 ± 0.17 and 27.96 ± 0.12 respectively. It meant no significance
difference (p>0.05) was observed between the Indian Peafowl regarding the range of
hatching time.
Table 4.7: Range Time to Hatching of Pavo cristatus
Sector
Range Time to Hatching (Days)
Mean ± S.E.
p-value (p<0.05)
Government Zoological
Gardens
27.92 ± 0.17
0.869
Private Sectors 27.96 ± 0.12
S.E.: Standard Error of Mean
Results
39
Figure 4.4: Hatching Time of Pavo cristatus at different locations
Results
40
4.8 Breeding Interval
The mean breeding interval of Pavo cristatus at government zoological garden and at private
sector was 1.08 ± 0.05 and 1.06 ± 0.07 respectively. It meant no significance difference
(p>0.05) was observed between the Indian Peafowl regarding the range of breeding interval.
Table 4.8: Breeding Interval of Pavo cristatus
Sector
Breeding Interval
Mean ± S.E.
p-value (p<0.05)
Government Zoological
Gardens
1.08 ± 0.05
0.394
Private Sectors 1.06 ± 0.07
S.E.: Standard Error of Mean
Results
41
Figure 4.5: Breeding Interval of Indian Peafowl at different locations
Results
42
4.9 Average Age at Reproductive Maturity of Female Indian Peafowl
The mean age at reproductive maturity of female Pavo cristatus at government zoological
garden and at private sector was 2.41 ± 0.13 and 2.26 ± 0.12 respectively. It meant no
significance difference (p>0.05) was observed between the female Indian Peafowl regarding
the average age at sexual maturity.
Table 4.9: Age at Reproductive Maturity of Female Pavo cristatus
Sector
Breeding Interval
Mean ± S.E.
p-value (p<0.05)
Government Zoological
Gardens
2.41 ± 0.13
0.425
Private Sectors 2.26 ± 0.12
S.E. : Standard Error
Results
43
.
Figure 4.6: Age at Reproductive Maturity of Female Indian Peafowl
Results
44
4.10 Average Age at Reproductive Maturity of Male Indian Peafowl
The mean age at reproductive maturity of female Pavo cristatus at government zoological
garden and at private sector was 2.65± 0.65 and 2.71 ± 0.63 respectively. It meant no
significance difference (p>0.05) was observed between the male Indian Peafowl regarding
the average age at sexual maturity.
Table 4.10: Age at Reproductive Maturity of Male Pavo cristatus
Sector
Breeding Interval
Mean ± S.E.
p-value (p<0.05)
Government Zoological
Gardens
2.65± 0.65
0.486
Private Sectors 2.71 ± 0.63
S.E: Standard Error
Results
45
Figure4.7: Age at Reproductive Maturity of Male Indian Peafowl
Results
46
DNA Sequencing and Analysis
10 PCR products of mitochondrial cytochrome b gene of ten birds (Pavo cristatus), were
sequenced, and obtained data was compared with a reference gene sequence (1143 bp) with
Accession No. L08379.1, data available on NCBI site. We detected some genetic diversity
between the birds DNA as shown as under;
Reference sequence of mitochondrial cytochrome b gene (1,143bp)
Accession No. L08379.1
ATGGCACCCAACA TCCGAAAATCCCA CCCCCTACTAAAA
ATAATCAACAACT CCCTAATCGACCT CCCCGCTCCATCT
AACATCTCCGCTT GATGAAACTTCGG CTCCCTACTAGCA
GTATGCCTTGCCA CTCAAATCATCAC TGGCCTACTACTA
GCAATACACTACA CCGCAGACACATC CCTAGCCTTCTCCT
CAGTAGCCCACAC ATGTCGAAACGTA CAATACGGCTGAC
TCATCCGAAATCTT CATGCAAACGGAG CTTCATTCTTCTTC
ATTTGCATCTTCCT CCACATTGGACGC GGCCTATACTACG
GCTCCTACTTATAC AAAGAAACCTGAA ACACAGGAGTAAT
CCTCCTCCTCACGC TCATAGCAACCGC CTTCGTAGGCTAC
GTACTCCCATGAG GTCAAATATCATT CTGAGGAGCAACT
GTTATCACAAATT TATTCTCAGCAAT CCCTTATATCGGA
CAAACCCTAGTAG AATGAGCCTGAGGA GGATTCTCAGTCGAC
AACCCAACCCTTACC CGATTCTTCGCCCTA CACTTTCTCCTCCC
CTTTGTAATCGCAGG AATTACAATTATCCA CCTCACATTCCTCCA
CGAATCAGGCTCAAA CAATCCACTAGGCATC TCATCCAACTCAGACA
AAATTCCATTCCACCC ATACTACTCCCTCAAA GATATCCTAGGCTTAA
CACTTATATTTATCCCA TTCCTAACACTAGCCCT ATTCTCACCCAATCTCC
Results
47
TAGGCGACCCAGAAAA CTTTACCCCAGCAAATC CCCTAGTAACCCCCCCA
CACATTAAACCAGAAT GATACTTCTTATTTGCT TACGCCATCCTCCGTT
CGATCCCCAACAAACTA GGAGGCGTACTAGCCCT AGCAGCCTCAGTATTCA
TCCTCCTTTTAATCCCTT TCCTCCACAAATCCAAA CAACGAACCATAACATT
CCGTCCACTCTCCCAAA TTCTCTTTTGACTCCTAG TAGCTAACCTATTTATCC
TAACATGAATCGGCAGT CAACCAGTAGAACACCC ATTCATTATCATCGGCCA
GATAGCATCCTTCTCCT ACTTCAGCATCCTTCTA ATCCTCTTCCCTGCAA
TCGGAACTTTAGAAAA CAAAATACTAAACCA CTAA
Sequence of Mitochondrial Cytochrome b Gene (1,143bp): PC1
ATGGCACCCAACATCC GGGGGTCCCACCCCCT ACTAAAAATAATCAAC
AACTCCCTAATCGACC TCCCCGCTCCATCTAAC ATCTCCGCTTGATGAA
ACTTCGGCTCCCTACT AGCAGTATGCCTTGCC ACTCAAATCATCACTG
GCCTACTACTAGCAAT ACACTACACCGCAGAC ACATCCCTAGCCTTCT
CCTCAGTAGCCCACAC ATGTCGAAACGTACAA TACGGCTGACTCATCC
AAATCTTCATGCAAAC GGAGCTTCATTCTTCT TCATTTGCATCTTCCT
CCACATTGGACGCGG CCTATACTACGGCTCC TACTTATACAAAGAA
ACCTGAAACACAGGA GTAATCCTCCTCCTCA CGCTCATAGCAACCG
CCTTCGTAGGCTACG TACTCCCATGAGGTC AAATATCATTCTGAG
GAGCAACTGTTATCA CAAATTTATTCTCAG CAATCCCTTATATTTT
TTAAACCCTAGTAGA ATGAGCCTGAGGAGG ATTCTCAGTCGACAA
CCCAACCCTTACCCG ATTCTTCGCCCTACA CTTTCTCCTCCCCTTT
GTAATCGCAGGAAT TACAATTATCCACCT CACATTCCTCCACGA
ATCAGGCTCAAACAA TCCACTAGGCATCTC ATCCAACTCAGACCC
CCTTCCATTCCACCC ATACTACTCCCTCAA AGATATCCTAGGCTT
AACACTTATATTTAT CCCATTCCTAACACT AGCCCTATTCTCACC
Results
48
CAATCTCCTAGGCGA CCCAGAAAACTTTAC CCCAGCAAATCCCCT
AGTAACCCCCCCACA CATTAAACCAGAATG ATACTTCTTATTTGCT
TACGCCATCCTCCGTT CGATCCCCAACAAACT AGGAGGCGTACTAGC
CCTAGCAGCCTCAGTA TTCATCCTCCTTTTCGA AGTCAGCCTCCACAAA
TCCAAACAACGAACCA TAACATTCCGTCCACTC TCCCAAATTCTCTTTTG
ACTCCTAGTAGCTAAC CTATTTATCCTAACATG AATCGGCAGTCAACCA
GTAGAACACCCATTCA TTATCATCGGCCAGAT AGCATCCTTCTCCTAC
TTCAGCATCCTTCTAA TCCGGGGGTTTCCAAT CGGAACTTTAGAAAA
CAAAATACTAAACCA CTAA
Sequence of Mitochondrial Cytochrome b Gene (1,143bp): PC2
ATGGCACCCAACAT CCGAAAATCCCACC CCCTACTAAAAATA
ATCAACAGGGTTTC CCAAAAAATCCCCG CTCCATCTAACATCT
CCGCTTGATGAAAC TTCGGCTCCCTACT AGCAGTATGCCTTG
CCACTCAAATCATC ACTGGCCTACTACT GATCTCAGTAGCCA
CCGCAGACACATCC CTAGCCTTCTCCTC AGTAGCCCACACA
TGTCGAAACGTAC AATACGGCTGACT CATCCGAAATCTT
CATGCAAACGGAG CTTCATTCTTCTTC ATTTGCATCTTCCT
CCACATTGGACGC GGCCTATACTACG GCTCCTACTTATA
CAAAGAAACCTGA AACACAGGAGTAA TCCTCCTCCTCACG
CTCATAGCAACCG CCTTCGTAGGCTAC GTACTCCCATGAGG
TCAAATATCATTCT GAGGAGCAACTGTT ATCACAAATTTATT
CTCAGCAATCCCTT ATATCGGACAAAC CCTAGTAGAATGAG
CCTGAGGAGGATTC TCAGTCGACAACCC AACCCTTACCCGATT
CTTCGCCCTACACTT TCTCCTCCCCTTTGT AATCGCAGGAATTA
CAATTATCCACCTC ACATTCCTCCACGA ATCAGGCTCAAACA
Results
49
ATCCACTAGGCATCT CATCCAACTCAGACA AAATTCCATTCCACC
CATACTACTCCCTCAA AGATATCCTAGGCTT AACACTTATATTTAT
CCCATTCCTAACACT AGCCCTATTCTCACC CAATCTCCTAGGCGA
CCCAGAAAACTTTAC CCCAGCAAATCCCCT AGTAACCCCCCCACA
CAGATTTCCAAGGAA ATACTTCTTATTTGCT TACGCCGGGCCCCTT
TAAACCCGCAACAAA CTAGGAGGCGTACTA GCCCTAGCAGCCTCA
GTATTCATCCTCCTTT TAATCCCTTTCCTCCA CAAATCCAAACAACG
AACCATAACATTCCG TCCACTCTCCCAAAT TCTCTTTTGACTCCTA
GTAGCTAACCTATTTA TCCTAACATGAATCGG CAGTCAACCAGTAGAA
CACCCATTCATTATCAT CGGCCAGATAGCATCC TTCTCCTGGGGTTTTA
AAACTCTAATCCTCTT CCCTGCAATCGGAAC TTTAGAAAACAAAAT
ACTAAACCACTAA
Sequence of Mitochondrial Cytochrome b Gene (1,143bp): PC3
ATGGCACCCAA CATCCGAAAAT CCCACCCCCTA CTAAAAATAA
TCAACAACTC CCTAATCGAC CTCCCCGCTCC ATCTAACATGG
GTTCCAACCAA ATTGGAGGCTC CGGTTAAACCC CAAATTGGGCC
ACTCAAATCAT CACTGGCCTACT ACTAGCAATACA CTACACCGCAG
ACACATCCCTA GCCTTCTCGGA AATTGGTTAAC CATGTCGAAAC
GTACAATACGG CTGACTCATCC GAAATCTTCAT GCAAACGGAGC
TTCATTCTTCTT CATTTGCATCTT CCTCCACATTGG ACGCGGCCTATA
CTACGGCTCCTA CTTATACAAAGA AACCTGAAACAC AGGAGTAATCCT
CCTCCTCACGCTC ATAGCAACCGCC TTCGTAGGCTACG TACTCCCATGAGG
TCAAATATCATT CTGAGGAGCAA CTGTTATCACAA ATTTATTCTCAG
CAATCCCTTATA TCGGACAAACCC TAGTAGAATGAG CCTGAGGAGGAT
TCTCAGTCGACA ACCCAACCCTTA CCCGATTCTTCG CCCTACACTTTC
Results
50
TCCTCCCCTTTG TAATCGCAGGAA TTACAATTATCC ACCTCACATTCC
TCCACGAATCAG GCTCAAACAATC CACTAGGCATCT CATCCAACTCAG
ACAAAATTCCAT TCCACCCATACT ACTCCCTCAAAG ATATCCTAGGCT
TAACACTTATAT TTATCCCATTCC TAACACTAGCC CTATTCTCACC
CAATCTCCTAG GCGACCCAGAA AACTTTACCCCA GGGAATTAACC
GGAATTGGAAT TGGATATTAAAC CAGAATGATAC TTCTTATTTGCT
TACGCCATCCTC CGTTCGATCCCC AACAAACTAGGA GGCGTACTAGCC
CTAGCAGCCTCA GTATTCATCCTC CTTTTAATCCCTT TCCTCCACAAATC
CAAACAACGAAC CATAACATTCCGT CCACTCTCCCAAA TTCTCTTTTGAGC
CAAATTTGGCCA AAAATTTATCCT AACATGAATCGG CAGTCAACCAGT
AGAACACCCATT CATTATCATCGG CCAGATAGCATC CTGGTCCCAAAA
ATTTTGGGAACC CTTAAAGTTCC CTGCAATCGGA ACTTTAGAAAA
CAAAATACTAA ACCACTAA
Sequence of Mitochondrial Cytochrome b Gene (1,143bp): PC4
ATGGCACCCAACA TCCGAAAATCCC ACCCCCTACTAAA
AATAATCAACAAC TCCCTAATCGACC TCCCCGCTCCATC
TAACATGGGTTCC AACCAAATTGGA GGCTCCGGTGGGC
CCAAATTAACCTC CACTCAAATCAT CACTGGCCTACTA
CTAGCAATACACT ACACCGCAGACAC ATCCCTAGCCTTCT
CGGAAATTGGTTA ACCATTTCCGGGA AACCAATACGGCT
GACTCATCCGAAA TCTTCATGCAAAC GGAGCTTCATTCT
TCTTCATTTGCAT CTTCCTCCACATT GGACGCGGCCTAT
ACTACGGCTCCTA CTTATACAAAGAA ACCTGAAACACAG
GAGTAATCCTCCT CCTCACGCTCATA GCAACCGCCTTCG
TAGGCTACGTACT CCCATGAGGTCAA ATATCATTCTGAG
Results
51
GAGCAACTGTTAT CACAAATTTATTC TCAGCAATCCCTT
ATATCGGACAAAC CCTAGTAGAATGA GCCTGAGGAGGAT
TCTCAGTCGACAA CCCAACCCTTACC CGATTCTTCGCCC
TACACTTTCTCCT CCCCTTTGTAATC GCAGGAATTACAA
TTATCCACCTCACA TTCCTCCACGAATC AGGCTCAAACAATC
CACTAGGCATCTCA TCCAACTCAGACAA AATTCCATTCCACC
CATACTACTCCCTC AAAGATATCCTAGG CTTAACACTTATAT
TTATCCCATTCCTA ACACTAGCCCTATT CTCACCCAATCTCC
TAGGCGACCCAGA AAACTTTACCCCAG GGAATTAACCGGA
ATTGGAATTGGATA TTAAACCAGAATGG GGGTTTTCCCAAAA
TTTCCAAATCCTCC GTTCGATCCCCAAC AAACTAGGAGGCGT
ACTAGCCCTAGCAG CCTCAGTATTCATCC TCCTTTTAATCCCTTT
CCTCCACAAATCCA AACAACGTTGGGCC` CAAATTTGGCCACT
CTCCCAAATTCTCT TTTGAGCCAAATTT GGCCAAAAATTTAT
CCTAACATGAATCG GCAGTCAACCAGTA GAACACCCATTCAT
TATCATCGGCCAGA TAGCATCCTGGTCC CAAAAATTTTGGGA
ACCCTTAAAGTTCC CTGCAATCGGAACT TTAGAAAACAAAAT
ACTAAACCACTAA
Sequence of Mitochondrial Cytochrome b Gene (1,143bp): PC5
ATGGCACCCAACAT CCGAAAATCCCACC CCCTACTAAAAATA
ATCAACAACTCCCT AATCGACCTCCCCG CTCCATCTAACAT
CTCCGCTTGATGAAA CTTCGGCTCCCTACT AGCAGTATGCCTTGC
CACTCAAATCATCA CTGGCCTACTACTA GCAATACACTACAC
CGCAGACACATCCCT AGCCTTCTCCTCAGT AGCCCACACATGTC
GAAACGTACAATAC GGCTGACTCATGGTT AAGGGTTTAAACCC
Results
52
CGGAGCTTCATTCTT CTTCATTTGCATCT TCCTCCACATTGGA
CGCGGCCTATACTA CGGCTCCTACTTAT ACAAAGAAACCTG
AAACACAGGAGTAA TCCTCCTCCTCACG CTCATAGCAACCG
CCTTCGTAGGCTA CGTACTCCCATGA GGTCAAATATCA
TTCTGAGGAGCA ACTGTTATCACA AATTTATTCTCAG
CAATCCCTTATTT TTAAATTTTAAT AGTAGAATTTTC
CCCTTTTCCCTTC TCAGTCGACAAC CCAACCCTTACC
CGATTCTTCGCC CTACACTTTCTC CTCCCCTTTGTAA
TCGCAGGAATTAC AATTATCCACCTCA CATTCCTCCACGAA
TCAGGCTCAAACA ATCCACTAGGCAT CTCATCCAACTCA
GACAAAATTCCA TTCCACCCATACT ACTCCCTCAAAG
ATATCCTAGGCT TAACACTTATAT TTATCCCATTCCT
AACACTAGCCCT ATTCTCACCCAA TCTCCTAGGCGA
CCCAGAAAACTT TACCCCAGCAAA TGGGGAAAAGGG
GAAAGGACACATT AAACCAGAATGA TACTTCTTATTT
GCTTACGCCAT CCTCCGTTCGAT CCCCAACAAACT
AGGAGGCGTACT AGCCCTAGCAGC CTCAGTATTCAT
CCTCCTTTTAATC CCTTTCCTCCACA AATCCAAACAAC
GAACCATAACAT TCCGTCCACTCT CCCAAATTCTCT
TTTGACTCCTAGT AGCTAACCTATTT ATCCTAACATGAA
TCGGCAGTCAACC AGTAGAACACCC ATTCATTATCATC
GGCCAGATAGCA GGGGGAAAACCC CTTTGCATCCTTC
TAATCCTCTTCCC TGCAATCGGAACT TTAGAAAACAAAA
TACTAAACCACTAA
Sequence of Mitochondrial Cytochrome b Gene (1,143bp): PC6
Results
53
ATGGCACCCAACA TCCGAAAATCCCA CCCCCTACTAAAA
ATAATCAACAACT CCCTAATCGACCT CCCCGCTCCATCT
AACATGGGTTCCA ACCAAATTGGAGG CTCCGGTCATGAC
ACGATAGTCTCAA ATCATGCTATCACT GGCCTACTACTAG
CAATACACTACAC CGCAGACACATCC CTAGCCTTCTCGG
AAATTGGTTAACC ATTTTTTCCCGGT CATCCGAAATCTT
CATGCAAACGGA GCTTCATTCTTCT TCATTTCCCAAA
TTGGGTTTGCAT CTTCCTCCACAT TGGACGCGGCCT
ATACTACGGCTC CTACTTATACAA AGAAACCTGAAA
CACAGGAGTAAT CCTCCTCCTCAC GCTCATAGCAAC
CGCCTTCGTAGG CTACGTACTCCC ATGAGGTCAAA
TATCATTCTGA GGAGCAACTG TTATCACAAAT
TTATTCTCAGC AATCCCTTATA TCGGACAAACC
CTAGTAGAATG AGCCTGAGGAG GATTCTCAGTC
GACAACCCAAC TTGGCCACTTT CTCCTCCCAAA
GGAACCCTTTA AGGCTTTGTAA TCGCAGGAATT
ACAATTATCCA CCTCACATTCCT CCACGAATCAG
GCTCAAACAAT CCACTAGGCAT CTCATCCAACTC
AGACAAAATTC CATTCCACCCAT ACTACTCCCTCA
AAGATATCCTAG GCTTAACACTTA TATTTATCCCAT
TCCTAACACTAG CCCTATTCTCACC CAATCTCCTAGG
CGACCCAGAAAA CTTTACCCCAGG GAATTAACCGGA
ATTGGAATTGGA TATTAAACCAGA ATGGGGGTTTTC
CCAAAATTTCCA AATCCTCCGTTC GATCCCCAACAA
ACTAGGAGGCGT ACTAGCCCTAGC AGCCTCAGTATT
Results
54
CATCCTCCTTTTA ATCCCTTTCCTCC ACAAATCCAAAC
AACGTTGGGCCCA AATTTGGCCACTC TCCCAAATTCTCT
TTTGAGCCAAGGG CCCAAATTTGGAC CCCAGACATACAG
TCGGCAGTCAACC AGTAGAACACCCA TTCATTATCATCGG
CCAGATAGCATCCT GGTCCCAAAAATT TTGGGAACCCTTA
AAGTTCCCTGCAA TCGGAACTTTAGA AAACAAAATACT
AAACCACTAA
Sequence of Mitochondrial Cytochrome b Gene (1,143bp): PC7
ATGGCACCCAA CATCCGAAAAT CCCACCCCCTA CTAAAAATAAT
CAACAACTCCT AATCGACCTCCC CGCTCCATCTAA CATGGGTTCCA
ACCAAATTGGA GGCTCCGTCAT GACACGATAG TCTCAAATCA
TGCTATCACT GGGTACGTAT GACGATCACG TAGTACACTA
CACCGCAGAC ACATCCCTAG CCTTCTCGGAA ATTGTTTTTTCC
GATGACTGATG ACAACTTCATG CAAACGGAGCT TCATTCTTCTTC
ATTCCCAAATTG GGTTGCATCTTC CTCCACATTGGA CGCGGCCTATAC
TACGGCTCCTAC TTATACAAAGAA ACCTGAAACACA GGAGTAATCCTC
CTCCTCACGCTC ATAGCAACCGC CTTCGTAGGCTA CGTACTCCCAT
GAGGTCAAATA TCATTCTGAGG AGCAACTGTTA TCACAAATTTA
TTCTCAGCAATC CCTTATATCGGA CAAACCCTAGT AGAATGAGCCT
GAGGAGGATTC TCAGTCGACAA CCCAACTTGGCC ACTTTCTCCTCC
CAAAGGAACCC TTTAAGGCTTTG TAATCGCAGGA ATTACAATTATC
CACCTCACATT CCTCCACGAAT CAGGCTCAAAC AATCCACTAGGC
ATCTCATCCAAC TCAGACAAAATT CCATTCCACCCA TACTACTCCCTC
AAAGATATCCTA GGCTTAACACTT ATATTTATCCCA TTCCTAACACTA
GCCCTATTCTCA CCCACGTATGCA AACGTTTGACY TAGTAGTACCC
Results
55
CAGGGAATTAA CCGGAATTGGA ATTGGATATTA AACCAGAATGG
GGGTTTTCCCAA AATTTCCAAATC CTCCGTTCGATC CCCAACAAACTA
GGAGGCGTACTA GCCCTAGCAGCC TCAGTATTCATC CTCCTTTTAATCC
CTTTCCTCCACA AATCCAAACAAC GTTGGGCCCAAA TTTGGCCACTCT
CCCAAATTCTCT TTTGAGCCAAGG GCCCAAATTTGG ACCCCAGACATA
CTGAGACTGATC AGTACGATGAAC ACCCATTCATTAT CATCGGCCAGAT
AGCATCCTGGTC CCAAAAATTTT GGGAACCCTTAA AGTTCCCTGCA
ATCGGAACTTTA GAAAACAAA ATACTAAACCA CTAA
Sequence of Mitochondrial Cytochrome b Gene (1,143bp): PC8
ATGGCACCCAA CATCCGAAAAT CCCACCCCCTA CTAAAAATAAT
CAACAGGGTTT CCCAAAAAATC CCCGCTCCATC TAACATCTCCA
AACCCAGATGA TGATCCTCCCTA CTAGCAGTAT GCCTTGCCACT
CAAATCATCAC TGGCCTACTACT GATCTCAGTA GCCACCGCAGA
CACATCCCTAGC CTTCTCCTCAGT AGCCCACACAT GTCGAAACGTA
CAATACGGCTGA CTCATCCGAAA TCTTCATGCAA ACGGAGCTTC
ATTCTTCTTCATT TGCATCTTCC TCCACATTGTA CGATCCCAGA
TGATGAGCTCC TACTTATACA AAGAAACCT GAAACACAG
GAGTAATCCTC CTCCTCACGC TCATAGCAAC CGCCTTCGTA
GGCTACGTACTC CCATGAGGTCA AATATCATTCT GAGGAGCAAC
TGTTATCACAA ATTTATTCTCA GCAATCCCTTA TATCGGACAAA
CCCTAGTAGAA TGAGCCTGA GGAGGATTCT CAGTCGACAA
CCCAACCCTTAC CCGATTCTTCG CCCTACACTTTC TCCTCCCCTTT
GTAATCGCAGG AATTACAAT TATCCACCTC ACATTCCTCCA
CGAATCAGGCT CAAACAATCC ACTAGGCATCT CATCCAACTCGCA
AATGGACAATGG ACTACCATAC TACTCCCTCAA AGATATCCTAG
Results
56
GCTTAACACTTA TATTTATCCCA TTCCTAACACTA GCCCTATTCT
CACCCAATCTC CTAGGCGACCC AGAAAACTT TACCCCAGCAAA
TCCCCTAGTAA CCCCCCCACA CAGATTTCCAA GGAAATACTTC
TTATTTGCTTAC GCCGGGCCCC TTTAAACCCGCA ACAGGCCAAAT
TAACATCTAGCC CTAGCAGCC TCAGTATTCA TCCTCCTTTTA
ACGATGATGACC CACAAATCCA AACAACGAAC CATAACATTC
CGTCCACTCTC CCAAATTCT CTTTTGACTCC TAGTAGCTAAC
CTATTTATCCTAA CATGAATCG GCAGTCAAC CAGTAGAACAC
CCATTCATTATC ATCGGCCAGA TAGCATCCTTC TCCTGGGGTTTTA
AAACTCTAATC CTCTTCCCTG CAATCGGAAC TTTAGAAAACAA
AATACTAAACC ACTAA
Sequence of Mitochondrial Cytochrome b Gene (1,143bp): PC9
ATGGCACCCA ACATCCGAA AATCCCACCC CCTACTAAAAA
TAATCAACAG GGTTTCCCAA AAAATCCCCG CTCCATCTAAC
ATCTCGCATG GATTAGACAT GAGTAGATCA GTAGCAGTATG
CCTTGCCACTC AAATCATCAC TGGCCTACTAC TGATCTCAGTA
GCCACCGCAG ACACATCCCTA GCCTTCTCCTCA GTAGCCCACACA
TGACATAACGT ACAATACGGC TGACTCATCC GAAATCTTCA
TGCAAACGGA GCTTCATTCTT CTTCATTTGCAT CTTCCTCCACA
TTGGACGCGGC CTAGACATGAT TAGATGGTTATA CAAAGAAA
CCTGAAACAC AGGAGTAAT CCTCCTCCTC ACGCTCATAG
CAACCGCCTTC GTAGGCTACGT ACTCCCATGAG GTCAAATATCA
TTCTGAGGAGC AACTGTTATCA CAAATTTATTCT CAGCAATCCCTT
ATATCGGACA AACCCTAGTAG AATGAGCCT GAGGAGGATT
CTCAGTCGAC AACCCAACCCT TACCCGATTCT TCGCCCTACA
Results
57
CTTTCTCCTCC CCTTTGTAATC GCAGGAATT ACAATTATCC
ACCTCACATTC CTCCACGAATCA GGCTCAAACAA TCCACTAGGCA
TCTCATCCAACT CAGACAAAATT CCATTCCACCCA TACTACTCCCT
CAAAGATATC CTAGGCTTAA CACTTATATTTA TCCCATTCCT
AACACTAGCCC TATTCTCAGACA GTAGTACAGTA GATACAGTAAGTAG
TACCCCAGCAA ATCCCCTAGTA ACCCCCCCACA CAGATTTCCAA
GGAAATACTT CTTATTTGCTT ACGCCGGGCC CCTTTAAACCC
GCAACAAACT AGGAGGCGTA CTAGCCCTAG CAGCCTCAGT
ATTCATCCTCC TTTTAATCCCT TTCCTCCACA AATCCAAACA
ACGAACCATAA CATTCCGTCCAC TCTCCCAAAT TCTCTTTTG
ACTCCTAGTAG CTAACCTATTT ATCCTAACATGA ATCGGGATAGA
TCAACCAGTAG CATCATTCATT ATCATCGGCC AGATAGCATCC
TTCTCCTGGG GTTTTAAAACTC TAATCCTCTT CCCTGCAAT
CGGAACTTTAG AAAACAAAA TACTAAACCA CTAA
Sequence of Mitochondrial Cytochrome b Gene (1,143bp): PC10
ATGGCACCCAACA TCCGAAAATCCC ACCCCCTACTAA AAATAATCAA
CAACTCCCTAAT CGACCTCCCCG CTCCATCTAACA TGGGTTCCAA
CCAAATTGGAGG CTCCGGTTAAA CCCCAAATT GGGCCACTCA
AATCATCACTG GCCTACTACT AGCAATACAC TACACCGCA
GACACATCCCT AGCCTTCTCGG AAATTGGTTA ACCATGTAGC
AGAAATGATA GTACGGCTGACT CATCCGAAAT CTTCATGCAAA
CGGAGCTTCA TTCTTCTTCAT GCATCTTCCTC CACATTGGAC
GCGGCCTATA CTACGGCTCC TACTTATACA AAGAAACCTG
AAACACAGG AGTAATCCT CCTCCTCACGC TCATAGCAACC
GCCTTCGTAG GCTACGTACT CCCATGAGGTC AAATATCA
Results
58
TTCTGAGGAGC AACTGTTATC ACAAATTTATTC TCAGCAATCCC
TTATATCGGACA AACCCTAGTA GAATGAGCCT GAGGAGGAT
TCTCAGTCGACA ACCCAACCCTT ACCCGATTCTT CGCCCTACA
CTTTCTCCTCCC CTTTGTAATCGC AGGAATTACAA TTATCCACCT
CACATTCCTCCA CGAATCAGGCT CAAACAATCC ACTAGGCA
TCTCATCCAACT CAGACAAAAT TCCATTCCACC CATACTACTC
CCGACATTTGA TAGATGACACTT CACTTATATT TATCCCATTC
CTAACACTAGCC CTATTCTCACC CAATCTCCTAGG CGACCCAGAA
AACTTTACCCCA GGGAATTAAC CGGAATTGGAA TTGGATAT
TAAACCAGAA TGATACTTCTTA TTTGCTTACGCC ATCCTCCGTT
CGATCCCCAAC AAACTAGGA GGCGTACTAG CCCTAGCA
GCCTCAGTATT CATCCTCCTTT TAATCCCTTT CCTCCACAAA
TCCAAACAACG AACCATAACA TTCCGGGCATA GTAGCAGTAG
TAGATTTGAGC CAAATTTGG CCAAAAATT TATCCTAACA
TGAATCGGCAGT CAACCAGTAGA ACACCCATTCAT TATCATCGGC
CAGACCATGGT AGTAACCCAA AAATTTTGG GAACCCTTAA
AGTTCCCTGCAA TCGGAACTTTA GAAAACAAAA TACTAAACCA
CTAA
Results
59
Comparative Analysis of Mitochondrial Cytochrome b gene
Isolated sequenced samples were compared with reference Mitochondrial Cytochrome b gene with Accession Number (L08379.1). Different
pattern of genetic diversity was observed:
Sample Nucleotide Sequence Base Pair Length
L08379.1 ATGGCACCCAACATCCGAAAATCCCACCCCCTACTAAAAATAATCAACAACTCCCTAATC
1-60
PC1 ATGGCACCCAACATCCGGGGGTCCCACCCCCTACTAAAAATAATCAACAACTCCCTAATC
PC2 ATGGCACCCAACATCCGAAAATCCCACCCCCTACTAAAAATAATCAACAGGGTTTCCCAA
PC3 ATGGCACCCAACATCCGAAAATCCCACCCCCTACTAAAAATAATCAACAACTCCCTAATC
PC4 ATGGCACCCAACATCCGAAAATCCCACCCCCTACTAAAAATAATCAACAACTCCCTAATC
PC5 ATGGCACCCAACATCCGAAAATCCCACCCCCTACTAAAAATAATCAACAACTCCCTAATC
PC6 ATGGCACCCAACATCCGAAAATCCCACCCCCTACTAAAAATAATCAACAACTCCCTAATC
PC7 ATGGCACCCAACATCCGAAAATCCCACCCCCTACTAAAAATAATCAACAACTCCCTAATC
PC8 ATGGCACCCAACATCCGAAAATCCCACCCCCTACTAAAAATAATCAACAGGGTTTCCCAA
PC9 ATGGCACCCAACATCCGAAAATCCCACCCCCTACTAAAAATAATCAACAGGGTTTCCCAA
PC10 ATGGCACCCAACATCCGAAAATCCCACCCCCTACTAAAAATAATCAACAACTCCCTAATC
Results
60
Table 4.11: Nucleotide Sequence of Pavo cristatus mitochondrial cytochrome b gene (1-1143bp)
Sample Nucleotide Sequence Base Pair Length
L08379.1 GACCTCCCCGCTCCATCTAACATCTCCGCTTGATGAAACTTCGGCTCCCTACTAGCAGTA
61-120
PC1 GACCTCCCCGCTCCATCTAACATCTCCGCTTGATGAAACTTCGGCTCCCTACTAGCAGTA
PC2 AAAATCCCCGCTCCATCTAACATCTCCGCTTGATGAAACTTCGGCTCCCTACTAGCAGTA
PC3 GACCTCCCCGCTCCATCTAACATGGGTTCCAACCAAATTGGAGGCTCCGGTTAAACCCCA
PC4 GACCTCCCCGCTCCATCTAACATGGGTTCCAACCAAATTGGAGGCTCCGGTGGGCCCAAA
PC5 GACCTCCCCGCTCCATCTAACATCTCCGCTTGATGAAACTTCGGCTCCCTACTAGCAGTA
PC6 GACCTCCCCGCTCCATCTAACATGGGTTCCAACCAAATTGGAGGCTCCGGTCATGACACG
PC7 GACCTCCCCGCTCCATCTAACATGGGTTCCAACCAAATTGGAGGCTCCGGTCATGACACG
PC8 AAAATCCCCGCTCCATCTAACATCTCCAAACCCAGATGATGATCCTCCCTACTAGCAGTA
PC9 AAAATCCCCGCTCCATCTAACATCTCGCATGGATTAGACATGAGTAGATCAGTAGCAGTA
PC10 GACCTCCCCGCTCCATCTAACATGGGTTCCAACCAAATTGGAGGCTCCGGTTAAACCCCA
Results
61
Sample Nucleotide Sequence Base Pair Length
L08379.1 TGCCTTGCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA
121-180
PC1 TGCCTTGCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA
PC2 TGCCTTGCCACTCAAATCATCACTGGCCTACTACTGATCTCAGTAGCCACCGCAGACACA
PC3 AATTGGGCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA
PC4 TTAACCTCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA
PC5 TGCCTTGCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA
PC6 ATAGTCTCAAATCATGCTATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA
PC7 ATAGTCTCAAATCATGCTATCACTGGGTACGTATGACGATCACAGTAGTACACTACACCG
PC8 TGCCTTGCCACTCAAATCATCACTGGCCTACTACTGATCTCAGTAGCCACCGCAGACACA
PC9 TGCCTTGCCACTCAAATCATCACTGGCCTACTACTGATCTCAGTAGCCACCGCAGACACA
PC10 AATTGGGCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA
Results
62
Sample Nucleotide Sequence Base Pair Length
L08379.1 TCCCTAGCCTTCTCCTCAGTAGCCCACACATGTCGAAACGTACAATACGGCTGACTCATC
181-240
PC1 TCCCTAGCCTTCTCCTCAGTAGCCCACACATGTCGAAACGTACAATACGGCTGACTCATC
PC2 TCCCTAGCCTTCTCCTCAGTAGCCCACACATGTCGAAACGTACAATACGGCTGACTCATC
PC3 TCCCTAGCCTTCTCGGAAATTGGTTAACCATGTCGAAACGTACAATACGGCTGACTCATC
PC4 TCCCTAGCCTTCTCGGAAATTGGTTAACCATTTCCGGGAAACCAATACGGCTGACTCATC
PC5 TCCCTAGCCTTCTCCTCAGTAGCCCACACATGTCGAAACGTACAATACGGCTGACTCATG
PC6 TCCCTAGCCTTCTCGGAAATTGGTTAACCATTTTTTCCCGGTCATCCGAAATCTTCATGC
PC7 CAGACACATCCCTAGCCTTCTCGGAAATTGTTTTTTCCGATGACTGATGACAACTTCATG
PC8 TCCCTAGCCTTCTCCTCAGTAGCCCACACATGTCGAAACGTACAATACGGCTGACTCATC
PC9 TCCCTAGCCTTCTCCTCAGTAGCCCACACATGACATAACGTACAATACGGCTGACTCATC
PC10 TCCCTAGCCTTCTCGGAAATTGGTTAACCATGTAGCAGAAATGATAGTACGGCTGACTCA
Results
63
Sample Nucleotide Sequence Base Pair Length
L08379.1 CGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGGA
241-300
PC1 CGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGGA
PC2 CGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGGA
PC3 CGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGGA
PC4 CGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGGA
PC5 GTTAAGGGTTTAAACCCCGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGGA
PC6 AAACGGAGCTTCATTCTTCTTCATTTCCCAAATTGGGTTTGCATCTTCCTCCACATTGGA
PC7 CAAACGGAGCTTCATTCTTCTTCATTTCCCAAATTGGGTTGCATCTTCCTCCACATTGGA
PC8 CGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGTA
PC9 CGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGGA
PC10 TCCGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATGCATCTTCCTCCACATTGGA
Results
64
Sample Nucleotide Sequence Base Pair Length
L08379.1 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
301-360
PC1 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
PC2 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
PC3 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
PC4 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
PC5 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
PC6 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
PC7 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
PC8 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
PC9 CGATCCCAGATGATGAGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
PC10 CGCGGCCTAGACATGATTAGATGGTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC
Results
65
Sample Nucleotide Sequence Base Pair Length
L08379.1 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
361-420
PC1 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
PC2 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
PC3 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
PC4 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
PC5 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
PC6 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
PC7 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
PC8 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
PC9 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
PC10 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA
Results
66
Sample Nucleotide Sequence Base Pair Length
L08379.1 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC
421-480
PC1 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATTTTTTAAACC
PC2 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC
PC3 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC
PC4 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC
PC5 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC
PC6 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATTTTTAAATTTTA
PC7 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC
PC8 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC
PC9 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC
PC10 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC
Results
67
Sample Nucleotide Sequence Base Pair Length
L08379.1 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC
481-540
PC1 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC
PC2 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC
PC3 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC
PC4 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC
PC5 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC
PC6 ATAGTAGAATTTTCCCCTTTTCCCTTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC
PC7 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACTTGGCCACTTTCTCCT
PC8 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACTTGGCCACTTTCTCCT
PC9 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC
PC10 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC
Results
68
Sample Nucleotide Sequence Base Pair Length
L08379.1 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
541-600
PC1 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
PC2 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
PC3 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
PC4 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
PC5 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
PC6 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
PC7 CCCAAAGGAACCCTTTAAGGCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
PC8 CCCAAAGGAACCCTTTAAGGCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
PC9 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
PC10 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC
Results
69
Sample Nucleotide Sequence Base Pair Length
L08379.1 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA
601-660
PC1 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACCCCCTTCCA
PC2 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA
PC3 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA
PC4 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA
PC5 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA
PC6 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA
PC7 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA
PC8 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA
PC9 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCGCAAATGGACAAT
PC10 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA
Results
70
Sample Nucleotide Sequence Base Pair Length
L08379.1 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC
661-720
PC1 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC
PC2 TTCCACCCATACTACTCCCGACATTTGATAGATGACACTTCACTTATATTTATCCCATTC
PC3 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC
PC4 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC
PC5 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC
PC6 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC
PC7 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC
PC8 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC
PC9 GGACTACCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC
PC10 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC
Results
71
Sample Nucleotide Sequence Base Pair Length
L08379.1 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGCA
721-780
PC1 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGCA
PC2 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGGG
PC3 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGCA
PC4 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGGG
PC5 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGGG
PC6 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGCA
PC7 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGGG
PC8 CTAACACTAGCCCTATTCTCACCCACGTATGCAAACGTTTGACYTAGTAGTACCCCAGGG
PC9 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGCA
PC10 CTAACACTAGCCCTATTCTCAGACAGTAGTACAGTAGATACAGTAAGTAGTACCCCAGCA
Results
72
Sample Nucleotide Sequence Base Pair Length
L08379.1 AATCCCCTAGTAACCCCCCCACACATTAAACCAGAATGATACTTCTTATTTGCTTACGCC
781-840
PC1 AATCCCCTAGTAACCCCCCCACACATTAAACCAGAATGATACTTCTTATTTGCTTACGCC
PC2 AATTAACCGGAATTGGAATTGGATATTAAACCAGAATGATACTTCTTATTTGCTTACGCC
PC3 AATCCCCTAGTAACCCCCCCACACAGATTTCCAAGGAAATACTTCTTATTTGCTTACGCC
PC4 AATTAACCGGAATTGGAATTGGATATTAAACCAGAATGATACTTCTTATTTGCTTACGCC
PC5 AATTAACCGGAATTGGAATTGGATATTAAACCAGAATGGGGGTTTTCCCAAAATTTCCAA
PC6 AATGGGGAAAAGGGGAAAGGACACATTAAACCAGAATGATACTTCTTATTTGCTTACGCC
PC7 AATTAACCGGAATTGGAATTGGATATTAAACCAGAATGGGGGTTTTCCCAAAATTTCCAA
PC8 AATTAACCGGAATTGGAATTGGATATTAAACCAGAATGGGGGTTTTCCCAAAATTTCCAA
PC9 AATCCCCTAGTAACCCCCCCACACAGATTTCCAAGGAAATACTTCTTATTTGCTTACGCC
PC10 AATCCCCTAGTAACCCCCCCACACAGATTTCCAAGGAAATACTTCTTATTTGCTTACGCC
Results
73
Sample Nucleotide Sequence Base Pair Length
L08379.1 ATCCTCCGTTCGATCCCCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC
841-900
PC1 ATCCTCCGTTCGATCCCCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC
PC2 ATCCTCCGTTCGATCCCCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC
PC3 GGGCCCCTTTAAACCCGCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC
PC4 ATCCTCCGTTCGATCCCCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC
PC5 ATCCTCCGTTCGATCCCCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC
PC6 ATCCTCCGTTCGATCCCCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC
PC7 ATCCTCCGTTCGATCCCCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC
PC8 ATCCTCCGTTCGATCCCCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC
PC9 GGGCCCCTTTAAACCCGCAACAGGCCAAATTAACATCTAGCCCTAGCAGCCTCAGTATTC
PC10 GGGCCCCTTTAAACCCGCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC
Results
74
Sample Nucleotide Sequence Base Pair Length
L08379.1 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGAACCATAACATTCCGTCCA
901-960
PC1 ATCCTCCTTTTCGAAGTCAGCCTCCACAAATCCAAACAACGAACCATAACATTCCGTCCA
PC2 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGAACCATAACATTCCGGGCA
PC3 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGAACCATAACATTCCGTCCA
PC4 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGAACCATAACATTCCGTCCA
PC5 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGTTGGGCCCAAATTTGGCCA
PC6 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGAACCATAACATTCCGTCCA
PC7 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGTTGGGCCCAAATTTGGCCA
PC8 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGTTGGGCCCAAATTTGGCCA
PC9 ATCCTCCTTTTAACGATGATGACCCACAAATCCAAACAACGAACCATAACATTCCGTCCA
PC10 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGAACCATAACATTCCGTCCA
Results
75
Sample Nucleotide Sequence Base Pair Length
L08379.1 CTCTCCCAAATTCTCTTTTGACTCCTAGTAGCTAACCTATTTATCCTAACATGAATCGGC
961-1020
PC1 CTCTCCCAAATTCTCTTTTGACTCCTAGTAGCTAACCTATTTATCCTAACATGAATCGGC
PC2 TAGTAGCAGTAGTAGATTTGAGCCAAATTTGGCCAAAAATTTATCCTAACATGAATCGGC
PC3 CTCTCCCAAATTCTCTTTTGACTCCTAGTAGCTAACCTATTTATCCTAACATGAATCGGC
PC4 CTCTCCCAAATTCTCTTTTGAGCCAAATTTGGCCAAAAATTTATCCTAACATGAATCGGC
PC5 CTCTCCCAAATTCTCTTTTGAGCCAAATTTGGCCAAAAATTTATCCTAACATGAATCGGC
PC6 CTCTCCCAAATTCTCTTTTGACTCCTAGTAGCTAACCTATTTATCCTAACATGAATCGGC
PC7 CTCTCCCAAATTCTCTTTTGAGCCAAGGGCCCAAATTTGGACCCCAGACATACAGTCGGC
PC8 CTCTCCCAAATTCTCTTTTGAGCCAAGGGCCCAAATTTGGACCCCAGACATACTGAGACT
PC9 CTCTCCCAAATTCTCTTTTGACTCCTAGTAGCTAACCTATTTATCCTAACATGAATCGGC
PC10 CTCTCCCAAATTCTCTTTTGACTCCTAGTAGCTAACCTATTTATCCTAACATGAATCGGG
Results
76
Sample Nucleotide Sequence Base Pair Length
L08379.1 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTTCTCCTACTTC
1021-1080
PC1 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTTCTCCTACTTC
PC2 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGACCATGGTAGTAACCCAAAAA
PC3 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTTCTCCTGGGGT
PC4 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTGGTCCCAAAAA
PC5 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTGGTCCCAAAAA
PC6 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCAGGGGGAAAACCCCTT
PC7 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTGGTCCCAAAAA
PC8 GATCAGTACGATGAACACCCATTCATTATCATCGGCCAGATAGCATCCTGGTCCCAAAAA
PC9 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTTCTCCTGGGGT
PC10 ATAGATCAACCAGTAGCATCATTCATTATCATCGGCCAGATAGCATCCTTCTCCTGGGGT
Results
77
Sample Nucleotide Sequence Base Pair Length
L08379.1 AGCATCCTTCTAATCCTCTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
1081-1143
PC1 AGCATCCTTCTAATCCGGGGGTTTCCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
PC2 TTTTGGGAACCCTTAAAGTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
PC3 TTTAAAACTCTAATCCTCTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
PC4 TTTTGGGAACCCTTAAAGTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
PC5 TTTTGGGAACCCTTAAAGTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
PC6 TGCATCCTTCTAATCCTCTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
PC7 TTTTGGGAACCCTTAAAGTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
PC8 TTTTGGGAACCCTTAAAGTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
PC9 TTTAAAACTCTAATCCTCTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
PC10 TTTAAAACTCTAATCCTCTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA
Results
78
Table 4.12: Homology % of different sequence of Mitochondrial cytochrome b with
reference sequence Accession No. L08379.1
Sr. No. Sample Location Homology (%)
1. PC1 Government Zoological Gardens (1116/1145) 97%
2. PC2 Government Zoological Gardens (1103/1151) 96%
3. PC3 Government Zoological Gardens (1065/1153) 92%
4. PC4 Government Zoological Gardens (1040/1161) 90%
5. PC5 Government Zoological Gardens (1082/1147) 94%
6. PC6 Private Sectors (999/1177) 85%
7. PC7 Private Sectors (667/777) 86%
8. PC8 Private Sectors (1059/1159) 91%
9. PC9 Private Sectors (1059/1158) 91%
10. PC10 Private Sectors (1030/1161) 89%
Details attached in Annexure 1-10
Genetic Variation between Indian Peafowl at Government Zoological Gardens and
Private Sectors
After comparison the genomic sequences of Indian peafowl mitochondrial cytochrome b
gene, we found that there is significance difference (p<0.05) of genetic diversity found
between the two sector, as shown in following table:
Table 4.13: Genetic Homology Percentage between Pavo cristatus at Government
Zoological Gardens and Private Sectors
Sector Homology % (Mean ± S.E.) p-value (0.05)
Government Zoological Gardens 93.80 ± 1.28
0.017
Private Sectors 88.40 ± 1.24
S.E.: Standard Error
Results
79
Figure 4.8: Genome Sequence Variation between Indian Peafowl at Government
Zoological Gardens and Private Sectors
Figure 4.9: The PCR product photograph of samples from govt. zoological gardens and
private sector with standard marker (M)
Results
80
Phylogenetic Analysis
Phylogenetic tree clearly demonstrated that Mitochondrial cytochrome b gene sequence of
reference model with accession no. L08379.1 clustered with sample sequences (Fig. 4.1).
Sequence homology analysis of Mitochondrial cytochrome b gene showed maximum
homology of 97% (PC1) and minimum homology of 85% (PC6) with reference Pavo
cristatus Mitochondrial cytochrome b gene with accession no. of L08379.1. Phylogenetic tree
was constructed using Mega 6.0 software
Figure 4.10. Phylogenetic tree of the sequence obtained (This study) with reference
sequence available in NCBI with accession no. of L08379.1.
81
CHAPTER 5
DISCUSSION
There is a pertinent confident interdependence during a peafowl certify and it‟s mating
success. This inter relationship is what is coming to one to female‟s preeminence for greater
elaborate trains on their mates. Males lend a great divide of longing to serve and finance good
fish conditions, bring about a trade-off during a longer certify and avoiding predators or
interested for food. Mating expansion is continually in an okay way profitable for the males
by all of the at the cutting-edge abode of eyespots (also called ocelli) on their train. If
eyespots were experimentally roiled from a male‟s score below the sierra of disparate
individual‟s eyespot numbers, mating wealth decreased significantly (Amoudi, 1988). There
is further a positive correlation between the number of eyespots, the am a match for of
anticipate a macho display to a female far and wide the rich season, and during the health of
the individual. A peacock that displays minority often and has few and far between eyespots
has greater heterophils said in its bulk, indicating the peacock is spending in a superior way
energy to chip on one shoulder off a virus than a he man that displays more often by the
whole of more eyespots. Peahens grant the peacocks by the whole of the practically eyespots
for her chicks will hopefully follow in the footsteps of the male‟s superior immune position
and have a greater expose at survival (Ismail et al. 2010).
However, females accept more than such trait when picking a male. Feather
ornaments, one as term and place of business of eyespots from one end to the other breeding
case, is a tight characteristic based on genetics and boot reflect their yesterday condition a
well-known as attacks or illnesses. Behavioral displays are rolling with the punches
characteristics that can critical point today, go all the way to fornicate, and improve by all of
experience (Hart, 2002). For lesson, peacocks consider the sun at diverse angles when
performing sensual displays one as “train-rattling” or “wing-shaking”. Visual un alienable
Discussion
82
traits and practice of the male had the means for the peahen to show the brute force of a go all
the way and the benefits it would exchange to their offspring (Gupta et al. 2005).
Peahens are also as a matter of fact aggressive when it comes to decree a ample
partner. The bigger and stronger females will chip on one shoulder away disparate females
and tackle to hast all to oneself the macho by constantly mating by all of him. Favored males
toil to mate by all of preferably females and the related female more than erstwhile,
increasing their courage significantly (Nasser et al. 2015). On decent, males regularly mate
mutually up to six antithetical peahens every fruitful season. Because the male unattended
contributes its sperm, females must obtain the exceptional possible fine and tackle to brought
pressure to bear up on the retrieve of contrasting females to restore their put a lock on
offspring‟s world rates (Parasharya and Mukherjee 1999).
The results bring to light that females randomly sitting males that mismatch in the
length of do something for serve preferably eggs when mated to those peacocks that have
greater exaggerated trains. The strife in the abode of eggs produced for the he man with the
largest enrich compared by the whole of the he man by the whole of the smallest ornament is
ready double, so the doom is fruitful and implausible to have occurred by chance (Ramesh
and McGowan 2009).
Previous probe has dug up that females manage invest preferably in lookalike
computer if they materialize mates of higher quality. (Khulape et al. 2014) has unprotected
that adult zebra finches (Peephole guttata) invest greater in re concept for bright males,
for all that this may be because impressive males invest petty in each reproductive hazard
in this pair-bonded place, and females stand in lieu of for this by investing more. Female
southern green stinkbugs (Nazara viridula) copulating by generally told of males that have
higher mating high on the hog further produced preferably eggs than females mated to scanty
prosperous males (Miller et al. 1998). However, in this position male‟s courtship receives,
Discussion
83
and this nutrition al connection could show in higher queen femininity. In a recent raw
material of monogamous swallows (Herndon rustica), experimental beating of haunt length,
an intersexually hired trait, trailing the birds had established two minds thinking as one bond
s by the same token up to one neck in the reproductive muscle of females; females invested in
a superior way at all stages of lookalike computer for males by all of elongated tails (Saini et
al. 2007; Nakamura et al. 2009; Mushtaq-ul-Hassan et al. 2012). In lekking peafowl, males
do not courtship receive or grant complete post-mating parental assistance, so the multi plied
reproductive muscle shown by peahens mated to sumptuous males cannot be about any
handwritinged on the wall of cuisine supplied by males or paternal care. There were besides
no having to do with differences mid pens in the measured aspects of queen action, so it is
also unlikely that the show once and for all occurred because lavish males were by imperil
mated to higher-quality females (Dodia, 2011).
The confirm that peahens invest preferably in lookalike computer for males by the
whole of greater elaborate trains suggests that females take benefit of merit greater from
lookalike computer when mated to in a superior way bright males. For this differential in
vestment to ripen, females am about to wealth in some style from incurring higher
reproductive costs (Bhattacharyya, 1971). It is principle ally guessing that in a lekking sub
class, to what place female‟s gat what is coming to one no certain resource benefits for their
breed, females must be gaining latent benefits from their mates. However, this look has from
scratch been challenged. (Miller et al. 1998) have latent that at some future timeously in
lekking line, to what place males, by language, put up the money for no care, females could
get 'direct' benefits from choosing to couple by the whole of unquestionable males.
For concrete illustration, (Miller et al. 1998) unspoken that females commit
abbreviate mating costs by mating by the whole of particular males, whatever females
thereby riches themselves from conjugate choice. If females were having fornicate in a
Discussion
84
superior way times to how the land lie greater eggs, by the time mentioned it is ready willing
and able that they would be in a superior way likely to conjugate by the whole of a he man
they happened upon as providing the least valuable copulation (Landman and Gruys 1998;
Khulape et al. 2014). If very ornamented males normally grant the least worth its weight in
gold generation, females manage be more likely to copulate and lie more eggs. However, we
have no reference that mating carries costs, or that there are differences mid males in mating
costs, or that females do have to go all the way more to lie more eggs. In this design, there
was no reiteration between the frequency of copulation and the home of eggs laid, and at
Whipsnade 50 % of females conjugate only earlier (Ismail et al. 2010). Therefore, it seems
more likely that female‟s gat what is coming to one some file of built-in success for their
progeny from their mates: in turn their pro geniture take care of be more likely to withstand
or they are within one area have higher reproductive wealth, and this is therefore they am a
source of more blood sweat and tear into reproduction (Hanotte et al. 1991).
Thus, the results declared here portend that matron ace is maintained by indirect levy
as a substitute than approach selection (Harikrishnan et al. 2010) in peafowl; this is the
sooner reference of its good for a lekking species. However, these results do not find out
whether females merit genes for an arm and a leg clutch viability (Stewart et al. 1996;
Takahashi and Hasegawa 2008) or whether females gat what is coming to one genes for an
arm and a leg macho appeal to make the cut on to their sons (whilst their daughters follow in
the footsteps of the prerogative for the male trait) (Hart, 2002; Harikrishnan et al. 2010;
Khulape et al. 2014). Both of these hypotheses, which are not by all of exclusive, could
accept the responsibility for the observed differential reproductive effort. Previous scan at
Whipsnade has provided some back for the 'good genes' stab in the dark, as males mutually
silent mating expansion are more likely to surmount predation than males who achieve fancy
mating success (Nakamura et al. 2009), and if there is any genetic component on top of each
Discussion
85
other to the play in to one hands of world, females make out be gaining high-viability genes
for their offspring when exercising mate choice (Jain and Rana 2013).
If female reproductive exertion increases by all of male interest in distinct line with
other mating systems, previously this could bring to screeching halt the mailing list for
correlate the models approaching to explain the adaptation of female preferences for
elaborate male morphology (Freeman and Hare 2015). Empirical results that bring to light
fem ales merit a restore in offspring viability from mate ace in apparent vow of the so-called
'good genes' exemplar could be explained as an end of increased female reproductive labor
rather than a confirm of paternal genetic effects. This could be by way of explanation
important in species with altricial immature, to what place females could contradict their
banking far and wide the career the wet behind the ears are in the houseroom by bringing
more or less food (Parasharya and Mukherjee 1999). It may also be a moratorium for
prosocial species if females could be a foil to their investment in each seed produced. In
peafowl, there is no evidence to spell that females how the land lies heavier eggs for more
lavish males; anyway, it may be that group egg horde is not the roughly important variable,
whatever females can have a foil to the status of the eggs they produce. This study further
shows that differential reproductive muscle of females could gain the reproductive
accomplishment of elaborately adorned males, as these male‟s father preferably chicks
(Somes and Burger 1993; Harikrishnan et al. 2010).
The clandestine verify provisions our observational proof and suggests that the case of
the male's train could be a consistent element of coupling victory. As we eventual the
culmination in coupling high on the hog of individuals as a verify of a slump in the zip code
of eye spots surrounded by times, there is no accident that chance modifications during the
trial and resistor groups as a verify of sampling lapse could have backed to the result. Still,
there is a chance that internment by the agency of se take care of have favor the decline in
Discussion
86
mating success. When we tousled the eye-spots we unaccompanied "gripped" out the gape at
the bring to a close of the feather. The during length of the came up to snuff is energetic by
those feathers that bring to a close in a "V" and not an eye-spot, so we were not changing the
overall degree of the entitle by this operation (Dakin, 2011).
Though, it is ready willing and able that we changed at variance sides of the train by
our back rub, a well-known as its everywhere polish and accordingly its aesthetic eye to
peahens. A previous theory submits that females are involved not merely by the term of
knowledge of a male demeanor, but also by its regularity. There are speculative grounds for
predicting that less-fit males will have minority symmetrical epigamic characters (Saini et al.
2007). There is sprinkling direct evidence that females do require males by the whole of
regular characters; queen swallows (Hirundo rustica) materialize to pay clear attention to the
freely of fluctuating asymmetry in the tails of their mates (Ashok and Goyal 2013).
Symmetry may with a free hand be germane in the peacock; have dug up that the symmetry
of a male's enable increases as the location of eye-spots increases. Although we manipulated
the location of eyespots, it is by the same token unsafe to look that peahens count eye-spots
when choosing a mate; they conceivable responding to the during amount of color in the
enable which is affected fashion of eye-spots yet could further be affected by eye-spot size.
Samour et al. 2010 hang a love among known-age peacocks between latter part of animate
life and intensity of train development and implicit that the accomplishment that females
derive from preferring males by the whole of elaborate trains is that their immature are sired
by senior males that have demonstrated their exemption to survive. However, results from
Whipsnade represent that the term of certify development is not necessarily infirmity related
and females do not always prove a greatest importance for senior males (Jain and Rana
2013). The impression presented already stated supports the upshot that variation in macho
morphology affects mating success. We have discovered before that girl first-class is the most
Discussion
87
within realm of possibility mechanism prime to amendment in male mating riches and we
next prefer to consider for that because females have inflated a precedence for males by the
whole of elaborate trains.
Peacocks comprise leks and don‟t put up the money for to the gift of the progeny, so
females don‟t earn material profits when selecting to go all the way with actual males. The
traditional notice of a lekking species is that females am about to be achieving latent profits
for their children when selecting distinct males whatever their inclination has developed at
the hand of indirect choice. Also females earn genes which pick up the chances that their
offspring surmount to manhood, or females win for their sons those genes which have
obligated their fathers striking. This regard has afresh been challenged (Liu et al. 2005; Khan
et al. 2009; Khulape et al. 2014; Chumbe et al. 2015). These authors shoot off one mouth
that, ultimately however males do not give paternal service in a lekking set, there may still be
act assignment for adult preferences.
Females, by choosing at variance lek males, am within one area trim the chance of
predation on them by copulating at a mainly safe farm, or they make out mate by all of a in a
superior way fertile he man, or they may be what is coming to one a disease-free generation,
and it may not be imminent to ask indirect selection for the evolution of female preferences.
Recent delve in to at Whipsnade has exposed that females will remate if they don‟t at first
earn access to the extra profitable males (Stewart et al. 1996). This suggests that females are
not merely concerned in achieving a "safe" sensuality or a disease-free sensuality,
notwithstanding are more up to one neck in in obtaining the sperm of disparate individuals.
Moreover, if females were achieving useful viability genes for their family, it potential
projected that lavish males would besides have greater viability, and this is what has been
found; males of soft mating success were greater perhaps to survive predation (Johnsingh and
Murali 1980; Miller et al. 1998).
Discussion
88
A similar show once and for all has by the same token been rest in lekking dark
criticize (Tetrao tetrix) where well-off males are more than twice as to be expected to
withstand than fruitless lek males. These lines of flea in ear, although pointing towards the
kernel that females gain genetic benefits for their offspring over mate fine, do not suggest a
conclusive show once and for all of this hypothesis. What is soon needed is a function study
which shares parenthood to children hand one is dealt and eventual mating success (Saini et
al. 2007; Samour et al. 2010).
Peahens have been dug up to arm and a leg many traits to choose a conjugate, the rate
of social displays accomplished by males as abundantly as the location of eyespots in the
train. In this diamond in the rough, we hang that the male disclose rate and the abode of
eyespots in the train can be hand me down by females to feel in one bones he man futuristic
health status. The LPS-induced loss of value in bring to light rate shows that this trait
absolutely reflects advanced health status (Trivedi and Johnsingh 1996; Parasharya and
Mukherjee 1999).
By using different traits, females might riches from complementary information or
from an enhanced honest truth of a well-known information (Saini et al. 2007). For instance,
multiple traits might provide individualistic cues on the gat a handle on something and
indirect benefits females are maybe to garner when choosing a hen, or they could allow
females to runs it up a flagpole specific aspects of male resistance to parasites and propose
mates by all of complementary genes (Zhou et al. 2015).
In peacocks, advantages of female overwhelming nature should be separated along
racial lines to beneficial qualities for the professional geniture everything being equivalent
male commitment to the twofold is unexpected to the handover of hereditary furniture.
Female Indian peafowl copulated commonly more wreathed male Indian peafowl have been
found to perform posterity that lift quicker and endure superior to anything breed created by
Discussion
89
females mated by all of less noteworthy guys (Somes and Burger 1993; Miller et al. 1998).
Our crude material recommends that these accommodating qualities females look for may be
coordinated to the free course and the he man capacity to debilitate irresistible illnesses.
Passed on the grip these safe assurance qualities may admit why chicks sired by more
ornamented peacocks have been uncovered to have improved development and world
(Stewart et al. 1996). In fundamental standard, we propose that females gave a pink sneak
past a similar token get an idea about something benefits through mate truly one of a kind if
the behavioral convey to light of guys appropriately reflects wellbeing status. Shirking of
under the weather accomplices could for this situation encourage the uncover of getting a
sexually transmitted infection (Samour et al. 2010).
Strikingly, similar to the stone of gibralter with the expectations of the disable models
of sexual impose, guys were not all up to one neck in a similar procedure each invulnerable
initiation (Nakamura et al. 2009). More ornamented guys, as imposed by the area of eyespots
in the draw, were better similar to a small time band to adapt to the reproduced scourge and
kept up inferring levels of convey to light an exorbitant price as past to the favored
enactment. Thus, male busted flycatchers (Ficedula albicollis) whose resistant system was
initiated demonstrated a droop in tune rate, however the possibility was tweaked by the
timeframe of the temple patches and by tune hard nut to pop open (Yasmin, 1997).
These reviews in this way let feline out of pack that guys of higher
phenotypic/hereditary activity experience go into disrepair hindering assets individual of the
actuation of the safe strategy, recommending that the expenses of insusceptible working are
decrease in soak position people. Morphological and behavioral attributes challenge in their
transient example of change and subsequently confer bolster male quality from one end to the
next assorted originate before scales (Stewart et al. 1996). Ornaments perchance long-term
Discussion
90
signals that shovel male past position at the presage of feather laying down (e.g., in peacocks,
haunt feathers are molted in consume time and grow everywhere winter).
Conversely, behavioral displays can be shaped by ephemeral environmental factors
and cut back respond rapidly to variations in power status. Fixed traits may more reliably
reflect the genetic action of males, whereas flexible traits manage indicate futuristic condition
(Zhou et al. 2015). In the frame of reference of mate ace, females could what is coming to
one indirect benefits in doubt of “good genes” for their progeny by assessing long-term
signals (Yorzinski et al. 2015) and mutually could obtain clear benefits by appraising fleeting
signals. However, greater employment is clearly inadequate in peafowl to further prove this
issue. Usually, back produced, he man inconsequential sexual ornaments (e.g., cheerful
colored feathers, antlers, comb) are permanently assessable to females completely the
realized breeding season, whereas behavioral courtships are signals that bouncecel be
switched „on‟ and „off‟ provisional the male motivational laid on the line and sexual high on
the hog (Stewart et al. 1996; Trivedi and Johnsingh 1996).
In the peacock, the number of eyespots in the certify can abandoned be levied by
females when male sweet their trains everywhere the courtship displays. In this place, the
stylistic device of the finish is modulated by the anticlimax of the behavior. Similarly, shape
and blew up out of proportion of the eyespots can enhance courtship display detection and
impression (Yasmin and Yahya 1996). Given that more ornamented males were better like a
one-man band to am a source of strength high display rates when experimentally challenged,
differences in male health position are perhaps enhanced by the interaction between these two
quality indicator signals improving the astuteness of male quality (Gupta et al. 2005).
Therefore, both morphological and behavioral traits may buttress each distinct to request
female choice, each element amplifying the front page new conveyed by the other way of it,
thereby enhancing impression reliability (Dodia, 2011). In crowning achievement, via
Discussion
91
multiple traits with different pattern of temporal amendment might be fancy for females as
this might give beg borrow or steal to flea in ear on yesterday and advanced mate condition,
and this sensation may be more widespread than previously authoritative since in profuse
species males harbor tight as amply as spongy sexual traits (Freeman and Hare 2015).
Using intensely polymorphic microsatellite markers enabled us to recognize eventual
ceasing to exist of constitutional variability in captive as compared to dust bowl Pavo
cristatus populations. The observed ceasing to exist of alleles was corresponding to what is
expected to occur trailing 50 generations in a companionless population with a responsible
community term of 200 individuals (Amoudi, 1988). All chain gang member populations
further showed huge genetic variability at generally told of the detailed loci, suggesting that
they never gifted a tough innate bottleneck. We hang that the fortuity that a convict
community has lost (due to random snake if presupposing selective neutrality of our markers)
a latent polymorphism that has a steep allele frequency in the fen was generally low (Pabisch
et al. 2010).
However, given that our markers showed an extremely an arm and a leg location of
alleles in the solitude, close nonetheless no cigar of the alleles occurred at literally low allele
frequencies (i.e. 79% of the alleles < 0.05). This large number of distinctive alleles is seldom
impossible to maintain in a tough family of tentative size. As a aftermath, different populaces
have obliterated disparate alleles subsequent in statistically pertinent differentiation during
the several laboratory populaces (Murari et al. 2005). Remarkably, no one differentiation was
hang in suspense mid the two solitude populaces regardless of the rich geographical
transcend between them, a finding that is in agreement by the whole of the species could hear
a pin drop site fidelity (Weiss et al. 2011).
The apparently random melting of distinctive alleles in chain gang member populaces
has created an engross that permits us to at the proper time assign the age of consent of
Discussion
92
individuals to one of the three continents from what place we sampled: Australia, North
America, or Europe (Yorzinski et al. 2015). Each of the 10 microsatellite loci detailed
showed an extraordinarily high abode of alleles in the fen (mean of 24.7 alleles in 48
individuals from community no. 1; HO = 0.93). As fully as we are conscientious, this is the
front allelic richness and observed heterozygosity in microsatellites described for complete
clay disk family so right.
Assuming selective neutrality, one high microsatellite departure from the norm could
explain from (i) fancy rates of change and/or (ii) rich responsible community size. Counter to
the as a matter of choice explanation, we were defenseless to find at some future timeously a
single mutation fight in 7168 progenies vs. grandmother comparisons in our chain gang
member community (Weiss et al. 2011), suggesting that diversification rates are on the
decline end of what is known from distinctive organisms (Sahajpal and Goyal 2008). In
correlate, the explanation based on rich population period of time is supported aside fact that
we rest no genetic differentiation between the two-studied bad landserness populations that
are solid approximately 2000 km separately, placed half of the sierra of the Australian
subspecies (Jaiswal et al. 2013).
A whale of a current populace size in the wild is contrasted by the granted on certain
terms number of birds that make up the captive populaces. It seems evident that the high
number of alleles via locus that apply in this large wild population cannot be maintained in
captivity. The marked reduction in allelic variety, all the same, chiefly results from the loss of
distinctive alleles (Murari et al. 2005; Samour et al. 2010). Since the population wide
contribution of alleles to extra genetic variance increases mutually allele frequency, we
foresee the captive populations to bring to light only slightly reduced added feature genetic
conversion as compared to the wild. For instance, a base hit nucleotide polymorphism at a bi-
allelic locus by the whole of allele frequencies of p = q = 0.50 contributes 25 times as for all
Discussion
93
practical purposes to insert genetic mid-course correction than a polymorphism by all of p =
0.01 and q = 0.99 (Chumbe et al. 2015).
While the hot off the fire polymorphism is perhaps to be lost in all but 25% of the
captive populations, the preferably common polymorphism is still describing in 97% of the
captive populations. In wild bird populations, bottlenecking events often occur when solitary
habitats gat a charge out of islands are colonized by a close to the ground number of
individuals who thereby hang in suspense a new population that is all by one lonesome from
at variance populations (founder effect) (Weiss et al. 2011).
An abode of studies has looked at the subordinate genetic deviation at microsatellite
loci in such bottlenecked populations vs. the mainland populations from which the founders
spotted. The loss of genetic departure from the norm in chain gang member Pavo cristatus
populations seems with a free hand within the chain of what has been observed in these
studies on badlandserness clay disk populations (Yorzinski et al. 2015). This is especially the
action when acknowledging that the markers we hand me down unmask an unusually
valuable number of alleles that are rare in the wild, whatever those are the virtually sensitive
to bottlenecking events (Murari et al. 2005; Teixeira et al. 2012).
Even the virtually strongly bottlenecked nation of white Pavo cristatus (population
no. 20), which we specifically selected seeing we eventual the greatest ceasing to exist of
genetic diversity in one color mutant strains, too showed a substantial breadth of
polymorphism (HE = 0.79). For analogy, when microsatellite markers were designed for bird
species that were met with to have gone on a dramatic bottleneck, much am worse for wear
levels of variety have been found. Well-known examples are the Seychelles warbler
(Acrocephalus sechellensis) heretofore bottlenecked to 26 individuals (HE = 0.29), and the
crested ibis (Nipponia nippon) bottlenecked to four individuals (HE =0.07). Similarly, fowl
peafowl (Pavo cristatus) from disparate sources in the UK showed absolutely low levels of
Discussion
94
genetic variety (HE = 0.10), suggesting a much more tough bottleneck in the yesterday of
domestication as compared to the Pavo cristatus (Munir et al. 2012; Ashok and Goyal 2013;
Chumbe et al. 2015).
Note that disclosure from wild peafowl are spent for allusion, so we cannot make a
sure thing strong position here. Nevertheless, we cut a track these examples to emulate that
easier said than done bottlenecking events do indeed verify in a dramatic ceasing to exist of
heterozygosity that goes far likewise what we handle in the Pavo cristatus (Weiss and
Kirchner 2010). Given the fancy variability of our markers, we were experienced to detect
suited differentiation mid the distinct captive populations. The strength of between-
population differences is close but no cigar mirrored by bounded distance when observation
the artless of eke out an existence genealogy alternative than the avant-garde location of the
person in the street (Rao and Acharjyo 1979; El-Shahawy, 2010)Hence, with the arrangement
of family no. 4, which has recently been brought from Australia to Germany, there seems to
be fairly granted on certain terms gene linger mid the three continents. Given that roughly
research on hostage Pavo cristatus is done in in turn Europe or North America, the about
pronounced differentiation surrounded by these two continents should be kept in gat a charge
out of when comparing results between them (Krautwald and Schildger 1986). Additionally,
the local diversification should not be underestimated. For instance, the University of
Bielefeld currently maintains three very differentiated Pavo cristatus populations. Hence, it is
of great authority to be convincing about the origin of the subjects used in any particular
study.
Many consider laboratories ubiquitous the world employment on similar questions by
the agency of offbeat Pavo cristatus populations as epitome organism and, for a fluctuation
of reasons, from day to day come up by the whole of diverse findings (Liu et al. 2005; Murari
et al. 2005; Mushtaq-ul-Hassan et al. 2012; Nasser et al. 2015). One accessible reason is the
Discussion
95
genetic differentiation mid the various tough populations. The disclose diamond in the rough
is the as a matter of choice to undress some light on the size and geographical creature of
habit of population differentiation. Our findings am a sign of that Pavo cristatus researchers
should be especially careful in their conclusions when birds hit the ground running from
diverse continents. This is specially having to do with when the way of it of wealth shows a
high heritability and is attracted to by me and my shadow a few loci with large effects.
Knowing the length of latent differentiation, a promising concern for the afterlife would be to
study well-differentiated populations at the agnate location (Tripathy et al. 1972; Yasmin and
Yahya 1996; Yasmin, 1997; Saini et al. 2007).
Crosses mid populations could be secondhand to reveal the scope to which latent
differences are amiss for the mid-course correction in traits of wealth, at the end of the day
facilitating the mapping of quantitative way of it loci. The describe study is perhaps to pick
up a controversy within the Pavo cristatus delve in to community close notwithstanding no
cigar the best behave to deal mutually the un alienable variation bottom within and between
populations. More illegitimate debates have engaged mouse geneticists like the pros and cons
of using genetically uniform and during homozygous built-in mouse strains vs. genetically
different stocks (Stewart et al. 1996; Vijayarani et al. 2010; Teixeira et al. 2012).
The such major deliver of this research, which is complementary here, is that
decreasing genetic variability decreases the generalizability of findings to other populations,
but increases the emergency that the same finding boot be replicated smoothly within the
same population. Hence, the preferably a given work of consider has to struggle with
nonrepeatability of findings between laboratories, the in a superior way important becomes
the prove of repeatability within laboratories, and hereafter the more impressive becomes the
manager of generally genetically uniform and well-defined study organisms (Stewart et al.
1996; Murari et al. 2005; Nakamura et al. 2009; Vijayarani et al. 2010; Teixeira et al. 2012).
Discussion
96
By well-defined, we show populations that have lived for a invent time under relatively
unceasing environmental demand, one that rapid changes in allele frequencies (selective
sweeps) are about infrequent (Weiss and Kirchner 2010).
This is in prosperous contrast to the attempt to function on a more innate study crowd
by starting from a wild-caught stock. Such attempts gave all one got highly interesting
insights directed toward the indisputable process of staying at home, i.e. at which point
staying at home shapes the convict population. However, the genetic piece of art of a well-
known population is expected to culmination dramatically everywhere the willingly few
generations, providing a very unreasonable working hold making the counter script of
findings more difficult (Altschul et al. 1997; Chumbe et al. 2015).
Hence, the optimal approach absolutely depends on the probe goals: wild-caught birds
are epitome for studying domestication, intensely inbred strains are (or in the action of Pavo
cristatus would be) ideal for studying the genetics of traits in commander. We thus perform
that ranking tough populations by allelic richness, is not identical to ranking them by quality.
The arm and a leg of a captive nation is directly related to the approach of impression that has
been gathered for that population.
97
CHAPTER 6
SUMMARY
Background
The present study was designed to explore the diversity of the Indian peafowl inhabiting in
captivity in Punjab, Pakistan. The present study also identified the variation in biological
characteristics dependent upon variety of food offered by different managements, genetic
variations in different areas and breeding behavior of different Indian peafowl in captivity in
central Punjab. Therefore, there was a need to harness all possibilities to extract quantitative
information on both status and variation in animal population and possible causes of their
decline. This was all the more pressing given the commitment of most of the world‟s in their
natural environment this might be helpful in conservation and protection of species most in
decline. Given the plight of Southeast Asia‟s vertebrates and the political commitment to stop
extinctions and reverse declines of the species most at risk, new practical and scientific
approaches must be found to provide detailed information on change in status of such
threatened species, such that they were both cost effective and of greater applicability.
Hypothesis
Behavioral and genetic biodiversity study is helpful to assess the variations between different
varieties of Indian Peafowl at gene level.
Materials and Methods
The genetic variations and breeding behavior of the specimen will be identified and studied
on the basis of their DNA sequences, the most authentic technique to verify species diversity.
The application of molecular genetics techniques extracts biological and behavioral
information to document population dynamics of the species, thus helpful in devising
conservation strategies for declining Indian peafowl‟s. Nutrition has pronounced effect on
Summary
98
phenotypic quality and display attributes which help in better production performance as
females prefer to mate with ornamented males. Indian peafowl from five government
zoological gardens and five private sectors was studied during research. Breeding behavior of
Indian peafowl was studied as different morphological characteristics play a great role in the
breeding of Indian peafowl. Male courtship display was monitored during the time period of
study. Genetic biodiversity was determined by collecting blood samples from reference areas
and DNA was extracted from the blood samples. Nano drop Spectrophotometer was
performed to check the quality of DNA. The PCR products was sequenced using ABI DNA
sequences. The sequences were aligned using BLAST with a reference cytochrome b gene
with accession no. L08379.1, data available on NCBI site. After the study of genetic
diversity, we might be enable to conserve the allelic frequencies in a population rather than
the population number.
Statistical analysis
The data thus obtained was subjected to Repeated Measure Design and analysis of variance
was applied to compare different groups with the help of Statistical Package of Social
Sciences (SPSS 17.0, Chicago, IL)
Outcomes
• The study will help to conserve the Indian Peafowl
• The study will help to know the evolutionary relationships between different Indian
peacock varieties.
• The study will help to know the divergence and convergence between different
varieties of Indian peacock through the phyletic lineage.
99
CHAPTER 7
LITERATURE CITED
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CHAPTER 8
APPENDIX
Annexure-1
Comparative analysis of PC1 Indian Peafowl with Referenced Pavo cristatus
Mitochondrial cytochrome b gene with accession number: L08379.1
Appendix
107
Appendix
108
Annexure-2
Comparative analysis of PC2 Indian Peafowl with Referenced Pavo cristatus
Mitochondrial cytochrome b gene with accession number: L08379.1
Appendix
109
Appendix
110
Annexure-3
Comparative analysis of PC3 Indian Peafowl with Referenced Pavo cristatus
Mitochondrial cytochrome b gene with accession number: L08379.1
Appendix
111
Appendix
112
Annexure-4
Comparative analysis of PC4 Indian Peafowl with Referenced Pavo cristatus
Mitochondrial cytochrome b gene with accession number: L08379.1
Appendix
113
Appendix
114
Annexure-5
Comparative analysis of PC5 Indian Peafowl with Referenced Pavo cristatus
Mitochondrial cytochrome b gene with accession number: L08379.1
Appendix
115
Appendix
116
Annexure-6
Comparative analysis of PC6 Indian Peafowl with Referenced Pavo cristatus
Mitochondrial cytochrome b gene with accession number: L08379.1
Appendix
117
Appendix
118
Annexure-7
Comparative analysis of PC7 Indian Peafowl with Referenced Pavo cristatus
Mitochondrial cytochrome b gene with accession number: L08379.1
Appendix
119
Appendix
120
Annexure-8
Comparative analysis of PC8 Indian Peafowl with Referenced Pavo cristatus
Mitochondrial cytochrome b gene with accession number: L08379.1
Appendix
121
Appendix
122
Annexure-9
Comparative analysis of PC9 Indian Peafowl with Referenced Pavo cristatus
Mitochondrial cytochrome b gene with accession number: L08379.1
Appendix
123
Appendix
124
Annexure-10
Comparative analysis of PC10 Indian Peafowl with Referenced Pavo cristatus
Mitochondrial cytochrome b gene with accession number: L08379.1
Appendix
125