UNIVERSITY OF VETERINARY & ANIMAL SCIENCES,...

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


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).


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


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


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


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


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


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


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


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


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).


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).


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


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


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,


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).


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.


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.


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).


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).


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


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


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)


Gardens

27.92 ± 0.17

0.869



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)


Gardens

1.08 ± 0.05

0.394



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)


Gardens

2.41 ± 0.13

0.425


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)


Gardens

2.65± 0.65

0.486


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


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


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


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


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


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


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


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


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


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








Results

60

Table 4.11: Nucleotide Sequence of Pavo cristatus mitochondrial cytochrome b gene (1-1143bp)


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


L08379.1 TGCCTTGCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA

121-180

PC1 TGCCTTGCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA

PC2 TGCCTTGCCACTCAAATCATCACTGGCCTACTACTGATCTCAGTAGCCACCGCAGACACA

PC3 AATTGGGCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA

PC4 TTAACCTCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA

PC5 TGCCTTGCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA

PC6 ATAGTCTCAAATCATGCTATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA

PC7 ATAGTCTCAAATCATGCTATCACTGGGTACGTATGACGATCACAGTAGTACACTACACCG



PC10 AATTGGGCCACTCAAATCATCACTGGCCTACTACTAGCAATACACTACACCGCAGACACA

Results

62


L08379.1 TCCCTAGCCTTCTCCTCAGTAGCCCACACATGTCGAAACGTACAATACGGCTGACTCATC

181-240

PC1 TCCCTAGCCTTCTCCTCAGTAGCCCACACATGTCGAAACGTACAATACGGCTGACTCATC


PC3 TCCCTAGCCTTCTCGGAAATTGGTTAACCATGTCGAAACGTACAATACGGCTGACTCATC

PC4 TCCCTAGCCTTCTCGGAAATTGGTTAACCATTTCCGGGAAACCAATACGGCTGACTCATC

PC5 TCCCTAGCCTTCTCCTCAGTAGCCCACACATGTCGAAACGTACAATACGGCTGACTCATG

PC6 TCCCTAGCCTTCTCGGAAATTGGTTAACCATTTTTTCCCGGTCATCCGAAATCTTCATGC

PC7 CAGACACATCCCTAGCCTTCTCGGAAATTGTTTTTTCCGATGACTGATGACAACTTCATG


PC9 TCCCTAGCCTTCTCCTCAGTAGCCCACACATGACATAACGTACAATACGGCTGACTCATC

PC10 TCCCTAGCCTTCTCGGAAATTGGTTAACCATGTAGCAGAAATGATAGTACGGCTGACTCA

Results

63


L08379.1 CGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGGA

241-300

PC1 CGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGGA




PC5 GTTAAGGGTTTAAACCCCGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGGA

PC6 AAACGGAGCTTCATTCTTCTTCATTTCCCAAATTGGGTTTGCATCTTCCTCCACATTGGA

PC7 CAAACGGAGCTTCATTCTTCTTCATTTCCCAAATTGGGTTGCATCTTCCTCCACATTGGA

PC8 CGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATTTGCATCTTCCTCCACATTGTA


PC10 TCCGAAATCTTCATGCAAACGGAGCTTCATTCTTCTTCATGCATCTTCCTCCACATTGGA

Results

64


L08379.1 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC

301-360

PC1 CGCGGCCTATACTACGGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC








PC9 CGATCCCAGATGATGAGCTCCTACTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC

PC10 CGCGGCCTAGACATGATTAGATGGTTATACAAAGAAACCTGAAACACAGGAGTAATCCTC

Results

65


L08379.1 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA

361-420

PC1 CTCCTCACGCTCATAGCAACCGCCTTCGTAGGCTACGTACTCCCATGAGGTCAAATATCA










Results

66


L08379.1 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC

421-480

PC1 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATTTTTTAAACC

PC2 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATATCGGACAAACC




PC6 TTCTGAGGAGCAACTGTTATCACAAATTTATTCTCAGCAATCCCTTATTTTTAAATTTTA





Results

67


L08379.1 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC

481-540

PC1 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC





PC6 ATAGTAGAATTTTCCCCTTTTCCCTTCTCAGTCGACAACCCAACCCTTACCCGATTCTTC

PC7 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACTTGGCCACTTTCTCCT

PC8 CTAGTAGAATGAGCCTGAGGAGGATTCTCAGTCGACAACCCAACTTGGCCACTTTCTCCT



Results

68


L08379.1 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC

541-600

PC1 GCCCTACACTTTCTCCTCCCCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC






PC7 CCCAAAGGAACCCTTTAAGGCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC

PC8 CCCAAAGGAACCCTTTAAGGCTTTGTAATCGCAGGAATTACAATTATCCACCTCACATTC



Results

69


L08379.1 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA

601-660

PC1 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACCCCCTTCCA

PC2 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCAGACAAAATTCCA







PC9 CTCCACGAATCAGGCTCAAACAATCCACTAGGCATCTCATCCAACTCGCAAATGGACAAT


Results

70


L08379.1 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC

661-720

PC1 TTCCACCCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC

PC2 TTCCACCCATACTACTCCCGACATTTGATAGATGACACTTCACTTATATTTATCCCATTC







PC9 GGACTACCATACTACTCCCTCAAAGATATCCTAGGCTTAACACTTATATTTATCCCATTC


Results

71


L08379.1 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGCA

721-780

PC1 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGCA

PC2 CTAACACTAGCCCTATTCTCACCCAATCTCCTAGGCGACCCAGAAAACTTTACCCCAGGG






PC8 CTAACACTAGCCCTATTCTCACCCACGTATGCAAACGTTTGACYTAGTAGTACCCCAGGG


PC10 CTAACACTAGCCCTATTCTCAGACAGTAGTACAGTAGATACAGTAAGTAGTACCCCAGCA

Results

72


L08379.1 AATCCCCTAGTAACCCCCCCACACATTAAACCAGAATGATACTTCTTATTTGCTTACGCC

781-840

PC1 AATCCCCTAGTAACCCCCCCACACATTAAACCAGAATGATACTTCTTATTTGCTTACGCC

PC2 AATTAACCGGAATTGGAATTGGATATTAAACCAGAATGATACTTCTTATTTGCTTACGCC

PC3 AATCCCCTAGTAACCCCCCCACACAGATTTCCAAGGAAATACTTCTTATTTGCTTACGCC

PC4 AATTAACCGGAATTGGAATTGGATATTAAACCAGAATGATACTTCTTATTTGCTTACGCC

PC5 AATTAACCGGAATTGGAATTGGATATTAAACCAGAATGGGGGTTTTCCCAAAATTTCCAA

PC6 AATGGGGAAAAGGGGAAAGGACACATTAAACCAGAATGATACTTCTTATTTGCTTACGCC





Results

73


L08379.1 ATCCTCCGTTCGATCCCCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC

841-900

PC1 ATCCTCCGTTCGATCCCCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC


PC3 GGGCCCCTTTAAACCCGCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC






PC9 GGGCCCCTTTAAACCCGCAACAGGCCAAATTAACATCTAGCCCTAGCAGCCTCAGTATTC

PC10 GGGCCCCTTTAAACCCGCAACAAACTAGGAGGCGTACTAGCCCTAGCAGCCTCAGTATTC

Results

74


L08379.1 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGAACCATAACATTCCGTCCA

901-960

PC1 ATCCTCCTTTTCGAAGTCAGCCTCCACAAATCCAAACAACGAACCATAACATTCCGTCCA

PC2 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGAACCATAACATTCCGGGCA

PC3 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGAACCATAACATTCCGTCCA


PC5 ATCCTCCTTTTAATCCCTTTCCTCCACAAATCCAAACAACGTTGGGCCCAAATTTGGCCA




PC9 ATCCTCCTTTTAACGATGATGACCCACAAATCCAAACAACGAACCATAACATTCCGTCCA


Results

75


L08379.1 CTCTCCCAAATTCTCTTTTGACTCCTAGTAGCTAACCTATTTATCCTAACATGAATCGGC

961-1020

PC1 CTCTCCCAAATTCTCTTTTGACTCCTAGTAGCTAACCTATTTATCCTAACATGAATCGGC

PC2 TAGTAGCAGTAGTAGATTTGAGCCAAATTTGGCCAAAAATTTATCCTAACATGAATCGGC


PC4 CTCTCCCAAATTCTCTTTTGAGCCAAATTTGGCCAAAAATTTATCCTAACATGAATCGGC

PC5 CTCTCCCAAATTCTCTTTTGAGCCAAATTTGGCCAAAAATTTATCCTAACATGAATCGGC


PC7 CTCTCCCAAATTCTCTTTTGAGCCAAGGGCCCAAATTTGGACCCCAGACATACAGTCGGC

PC8 CTCTCCCAAATTCTCTTTTGAGCCAAGGGCCCAAATTTGGACCCCAGACATACTGAGACT


PC10 CTCTCCCAAATTCTCTTTTGACTCCTAGTAGCTAACCTATTTATCCTAACATGAATCGGG

Results

76


L08379.1 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTTCTCCTACTTC

1021-1080

PC1 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTTCTCCTACTTC

PC2 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGACCATGGTAGTAACCCAAAAA

PC3 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTTCTCCTGGGGT

PC4 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTGGTCCCAAAAA


PC6 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCAGGGGGAAAACCCCTT


PC8 GATCAGTACGATGAACACCCATTCATTATCATCGGCCAGATAGCATCCTGGTCCCAAAAA

PC9 AGTCAACCAGTAGAACACCCATTCATTATCATCGGCCAGATAGCATCCTTCTCCTGGGGT

PC10 ATAGATCAACCAGTAGCATCATTCATTATCATCGGCCAGATAGCATCCTTCTCCTGGGGT

Results

77


L08379.1 AGCATCCTTCTAATCCTCTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA

1081-1143

PC1 AGCATCCTTCTAATCCGGGGGTTTCCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA

PC2 TTTTGGGAACCCTTAAAGTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA

PC3 TTTAAAACTCTAATCCTCTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA



PC6 TGCATCCTTCTAATCCTCTTCCCTGCAATCGGAACTTTAGAAAACAAAATACTAAACCACTAA





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%





6. PC6 Private Sectors (999/1177) 85%





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


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.


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

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106

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

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Annexure-2



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