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reproductive physiology in queen (cat)
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Asst. Prof. Hala Jawad Alfatlawy

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Study of some Anatomical,
Physiological and Pathological
aspects of feral Queen’s
Reproductive System in Iraq
A Thesis Submitted
To
The Council of the College of Veterinary Medicine ,
University of Al-Qadisiya
in Partial Fulfillment of The Requirement for
the Degree of Master of Science in
Theriogenology
By
Hella Jewad Kadhim Al-Fatlawy
B.V.M.S
Supervised By
Asst. Prof. Dhia H.Jassim, D.Sc Asst.Prof. Thaier A.Abid,
D.Sc
2010 A.D.
1431 A.H.
Supervisors Confirmation
We certify that this thesis was prepared under our
supervision at the Department of Surgery and
Obstetrics, College of Veterinary Medicine/
University of Al-Qadisiya in partial fulfillment of
the requirement for the degree of master in
Theriogenology
Supervisor Supervisor
Asst .Prof. Dhia H.Jassim,D.Sc Asst. Prof. Thaier A.Abid,

D.Sc
/ /2010 / /2010
In the view of the available recommendations I
forward this thesis for debate by the examination
committee
Asst .Prof. Jabbar A.A. AL-Saaidi , D.Sc

Vice Dean for Postgraduate Studies
/ /2010
We, the members of the examining committee ,certify that after reading
this thesis and examining the student in the contents, we think it is
adequate for the award of the master degree of veterinary medicine /
Theriogenology.
Dr. Mohammad J. Alwan
Prof. (Chairman)
College of Vet. Med., University of Baghdad
Dr.Ihsan Hammody Saeed Dr. Taher Abdl- Hussain
Asst. Prof.(member) Asst. Prof.(member)
College of Vet. Med., College of Vet. Med.,
University of Baghdad University of Basra

Dr. Dhia ,H. Jassim Dr. Thaier A. Abid
Asst. Prof.(member and supervisor) Asst. Prof.(member and

supervisor)
College of Vet. Med., College of Vet. Med.,
AL-Qadisiya university AL-Qadisiya university
Approved by the council of the college of the Veterinary Medicine
Asst.Prof. Dhia H. Jassim Prof. Dr. Khairi

A.Dawood
Head of department the Dean
Date of Examination : 27/6/2010
Dedication
To ..My Beloved Husband ,Dr. Haider Al-

Ramahi
For His Encouragement and
Patience
To …My Sweetie Daughter, Wid
For Her Lovely Smile
Hella
Acknowledgement
Forefront ,most I thank the great God, who gives me strength and
patience to carry out this study.
I would like to express my extra thanks to my supervisor Dr.
Dhia Hussein and Dr.Thaeir Alwan for their help and advices to
complete this thesis. Many thanks, also to Prof. Dr. Khairi A.
Dawood, Dean of College of Vet. medicine for his generous
supports .Also, I would like to thank Assist. Prof. Dr. Jabbar A. Al-
Saaidi, Head of post graduate studies committee for his support
and help.
Many thanks to the Head and all staff of the department of
Veterinary Surgery and Obstetrics, Vet. Medicine College of Al-
Qadisiya University for their kind assistance, and to Dr. Khaleel,
G. for microscopical photography .
My great gratitude to Dr.Hassan Al-Awadi, Dean of Vet.
Medicine college of Babylon University, Head and staff of the
department of Public Health in same College, Assist.Prof. Mujeeb
Shamsa (An najaf technical institute) ,Dr. Helmee Hamid (College
of Vet.Med. Babylon Univ.), Prof. Dr.Muhammad Jewaed and the
staff of the department of Pathology-College of Vet. Med.
Baghdad Univ. and the staff of diagnostic laboratory in Al-
Diwanyia general hospital .
All thanks to the staffs of libraries of Vet. Med. Colleges of Al
Qadisiya , Babylon and Baghdad Universities. Finally I would like
to thank any person who help to complete this thesis.
Hella J.Khadim
Abstract
The present study is carried out to investigate some aspects of
reproduction in Iraqi feral cats. Sixty blood samples and female genital
specimens from sexually mature queens were trapped during the period
from December 2008 to December 2009.The specimens were classified into
pregnant and non pregnant. The non pregnant were further classified into
reproductive systems with or without obvious pathological changes.
During the entire study period, the results showed that the means of length
,width and thickness of right and left ovaries were 9.8 × 5.03 ×3.95 mm and
9.56 ×4.8 ×3.05 mm respectively, with no significant variation (p≥ 0.05)
between the dimensions of right and left ovaries or between dimensions of
the different seasons. The means of length of right and left uterine tubes
were 54.4 and 55.7 mm respectively ,with no significant variation (p≥ 0.05)
between the dimensions of right and left uterine tubes or between dimension
of different seasons. The means of length and diameter of right and left
uterine horns were 48.44 × 5.41 mm and 54.43 × 5.14mm ,with significant
variation (P≤0.01) between dimensions of right and left uterine horns and
between the dimensions of different seasons .Also, the result showed that the
means of the length and diameter of uterine bodies and cervix were 22.75
×5.29 mm and13.2 × 8.72mm respectively with significant variation
(P≤0.01) between dimensions of different seasons .
The physiological study results showed that incidence rate of follicular
phase was 24.56 % and the highest incidence was in Winter 62.5% followed
by Spring and Summer in percentage 44.4% and 20% respectively, while no
follicular activity was noted in animals during Autumn . The hormonal assay
showed that the mean of the estradiol -17ß in 14 follicular phase animals
was 47.5pg/ml, while The percentage of vaginal epithelial cells in 14 estrus
queens for parabasal cells, intermediate cells nucleated superficial cells and
anucleated superficial cells were 0.92% , 12.21 % , 61.5 % and 27.71%
respectively.
In present study the luteal phase incidence was 29.82% and the highest
incidence was in Spring 56.66%,followed by Summer and Autumn in
percentage 41% and 20% respectively, while it was 6.66% in Winter. The
average of corpora lutea per animal was 4.7 % and the incidence rate of
pregnant queens in this study was 19.29%,the highest incidence was in
Summer 33.33%,followed by Spring and Autumn in incidence rate 28.57%
and 25% respectively, while no pregnant cat was captured in Winter. The
implantation rate was 86.06% and the transuterine migration of embryos was
observed in incidence rate 27.27% and all transuterine migration of embryos
was occurred in animals with 100% implantation rate . The maximum
number of embryos that migrate was 2 /animal. The incidence rate of
pseudopregnancy 11.76%, and characterized by present of active corpora
lutea on ovaries , with slight enlargement of uterus and uterine tube .
In all animals with diestrus the mean of plasma progesterone level was 48
ng/ml ,and the cell comprising the vaginal smear include intermediate cells
47% parabasal cells45% and nucleated cells 5% .
The anestrus phase was recognized in incidence rate 36.84% and the
highest incidence of anestrus phase was noted in Autumn 75%, followed by
Winter ,Summer and Spring in incidence rate 46.66% ,20% and 7.14%
respectively. Cells comprising the vaginal smear in anestrus queens include
intermediate cells 87.9%, parabasal cells 8.6%,nucleated superficial cells
2.7% , anucleated superficial cells 0.8% .
The pathological study was revealed 3 out of 60 queens with gross
reproductive systems abnormalities ,with incidence rate 5% ,these cases
include paraovarian cyst, follicular cysts and pyometra.
Content
No. Title Page
Chapter One: Introduction
1 introduction 1-2
Chapter Two: Review of Literatures
2 Review of Literatures 3-36
2.1. The Cats 3
2.1.1. Scientific Classification of The Domestic Cats 3
2.1.2. Domestication and History 4
2.1.3. Hybridation with Wild Cats 4
2.1.4. Breeds 5
2.2. Feline Reproduction 6
2.2.1. Anatomy of Female Genital Tract 6
2.2.1.2 Ovaries 6
2.2.1.2.1. Morphology and Location 6
2.2.1.2.2. Blood Supply 6
2.2.1.2.3. Ovarian Follicles 7
2.2.1.2.4. Corpous Luteum 7
2.2.1.3 Uterine Tubes (oviducts) 8
2.2.1.3.1 Morphology and Location 8
2.2.1.4. Uterus and Cervix 8
2.2.1.4.1 Morphology and Location 8
2.2.1.4.2. Blood Supply 9

2.2.1.5. Vagina and Vestibule 9
2.2.1.5. 1. Blood Supply 10
2.2.1.6. Vulva and Clitoris 10
2.2.2. Physiology of Female Reproduction 11
2.2.2.1. Breeding Season 11
2.2.2.2. Puberty 13
2.2.2.3. Estrous Cycle 14
2.2.2.4 Phases of Estrous Cycle 15
2.2.2.4.1. Proestrus 15
2.2.2.4.1.1 Definition 15
2.2.2.4.1.2. Duration and Signs 16
2.2.2.4.1.3. Hormonal Changes 16
2.2.2.4.1.4. Vaginal Cytology 16
2.2.2.4.2. Estrus (Follicular Phase) 17
2.2.2.4.2.2 Duration and Signs 17
2.2.2.4.2.3. Horminal Changes 18
2.2.2.4.3. Interestrus 19
2.2.2.4.3.1. Definition 19
.2.2.4.3.2. Duration and Signs 20
2.2.2.4.4. Anestrus 21
2.2.2.4.4.2 Duration and Signs 22
2.2.2.4.5. Dioestrus 23
2.2.2.4.5.1. Definition 23
2.2.2.4.5.2. Duration & Clinical Signs 24
2.2.2.5. Mating and Ovulation 25
2.2.2.6.4. Hormonal Changes 28

2.2.2.7. Pregnancy 28
2.2.2.7.1. Definition 28
2.2.2.7.3. Behavior 29
2.2.2.7.4. Hormonal Changes 29
2.2.3 Pathology of Female Reproductive System 30
2.2.3.1. Disorders of The Ovary 30
2.2.3.1.1. Ovarian and Parovarian Cysts 30
2.2.3.1.1.1 Cystic Rete Ovarii 30
2.2.3.1.2.2. Follicular Cysts 30
2.2.3.1.2.3. Serosal Inclusion Cysts 31
2.2.3.1.2.4. Parovarian (Periovarian) Cysts 31
2.2.3.1.3. Ectopic Adrenocortical Parovarian Nodules 31
2.2.3.1.4. Ovarian Neoplasia 31
2.2.3.2. Disorders of the Uterine Tube 32
2.2.3.2.1. Hyperplasia of the Uterine Tube 32
2.2.3.2.2. Salpingitis 33
2.2.3.3. Disorders of The Uterus 33
2.2.3.3.1. Cystic Endometrial Hyperplasia 33
2.2.3.3.2. Endometritis and Pyometra 34
2.2.3.3.4. Acute Metritis 34
2.2.3.4. Disorders of queen’s Vagina 36
2.2.3.4.1. Vaginal Prolapse 36
2.2.3.4.2. Vaginitis 36
2.2.3.4.3 Vaginal Neoplasia 36
Chapter Three: Materials and Methods
3. Materials and Methods 37-48
3.1. Materials 37
3.1.1. Animals 37
3.1.2. Sampling 38
3.1.2.1. Blood Samples 38
3.1.2.2. Vaginal Smears 38
3.1.2.3. Female Reproductive Tract Samples 38

3.1.3. Culture media 39
3.1.4. Reagents and Solutions 39
3.1.5. Chemicals 40
3.1.6. The Diagnostic Equipments 40
3.1.6. Different Materials 40
3.2. Methods 40
3.2.1. Designing of Study 40
3.2.1.1. Non Pregnant Reproductive Systems Samples 43
3.2.1.2. Pregnant Reproductive Systems Samples 43
3.2.1.3. Measurement of Reproductive Tract 44
3.2.1.4. Hormonal Examination 44
3.2.1.5. Bacterial Isolation 45
3.2.1.5. 1 Preparation and Sterilization of Media 45
3.2.1.5.2. Preparation of Reagents 45
3.2.1.5.3. Isolation of Bacteria 45
3.2.1.5.4. Identification of Isolates 46
3.2.1.5.5. Bacterial Identification by APi-20E 48
3.2.1.5.6. Statistical Analysis 48
Chapter Four : Results
4. Results 49
4.1. Anatomical Study 49
4.1.1. Topography of Queen’s Reproductive System 49
4.1.1.1. Ovary 49
4.1.1.2. Uterine Tube 49
4.1.1.3 Uterus 49
4.1.2. Measurments of Queen’s Reproductive System 54
4.1.2.1. Ovary 54
4.1.2.2. Uterine Tube 55
4.1.2.3. Uterine Horn 58
4.1.2.4. Uterine Body 60
4.1.2.5. Cervix 61
4.2. Physiological study 63
4.2.1. Follicular Phase 63
4.2.2. Luteal phase 68
4.2.3. Anestrus phase 76
4.3. Pathological Study 80
Chapter Five : Discussion
5. Discussion 87
5.1. Anatomical Study 87
5.1.1. Topography of Genital Parts:
5.2. Physiological Study 89
5. 2.1. Incidence of Sexual Activity (Follicular and Luteal Phases) 89
5.2.2 Incidence of Sexual Inactivity (Anestrous Phase) 90
5.2.3. Pregnancy and Pseudopregnancy 91
5.2.6 Vaginal Cytology 92
5.2.7. Hormonal Changes 93
5.3. Pathological Study 94
Chapter Six : conclusions and recommendation
6.1 Conclusions 96
6.2 Recommendations 97
Chapter Seven : References
7 References 98-113
List of Tables
No. Title page
2.1 Scientific Classification of Domestic Cat 3
2.2 The Domestic Cat's Breeds 5
2.3 The Vaginl Cytology in Interestrus Phase 21
2.4 The Vaginl Cytology in Anestrus Phase 23
2.5 The Vaginl Cytology in Dioestrus Phase 25
2.6 Some of Disorders of Queen’s Ovary 32
2.7 Some of Disorders of Queen’s Uterus 35
4.1 The Measurement of Ovaries During Different Seasons 56
4.2 The Measurements of Ovaries According to Age 57
4.3 The Measurements of Uterine Tubes During Different Seasons 57
4.4 The Measurements of Uterine Tubes According to Age 58
4.5 The Measurements Uterine Horns During Different Seasons 59
4.6 The Measurement Uterine Horns According to Age 60
4.7 The Measurements of Uterine Body According to Season 61
4.8 The Measurements of Uterine Body According to Age 62
4.9 The Measurements of Cervix According Season 62
4.10 The Measurements of Cervix According to Age 62
4.11 The Incidence of Follicular Phase According to Individual Ovary and 65
Season
4.12 The Relationship Between Number of C. L. and No. of Fetuses 72
4.13 the measurements of reproductive system with parovarian cyst 81
4.14 the measurement of the reproductive system with follicular cyst 83
List of Figures
No. Title page
2.1 The Estrus Cycle of Domestic Queen 15
3.1 The Local Made Feral Cat Trap 37
3.2 The Steps of Study Design 42
4.1 The Position of Ovary, Caudal to Kidney 50
4.2 The Suspensory Ligament 50
4.3 The Proper Ligament 51
4.4 The Ovarian Bursa 51
4.5 The Topography of Uterine Tube 52
4.6 The Unbend Uterine Tube 52
4.7 The Shape of The Uterus 53
4.8 The Uterine Body 53
4.9 The Broad Ligament and The Round Ligament 54
4.10 The Incidence of Follicular Phase in Different Seasons 63
4.11 The Present of 2Follicles on Left Ovary 64
4.12 The Incidence of Follicular Phase in Study Months 64
4.13 The Incidence of Follicular Phase According to Individual Ovary 65
and Season
4.14 The Straight Uuterus in Follicular Phase 66
4.15 The Spiral Uterus in Follicular Phase 67
4.16 The Vaginal Cytology of Follicular Phase 67
4.17 The Incidence of Luteal Phase in Different Seasons 68
4.18 The Incidence of Luteal Phase in Study Months 69
4.19 Corpus Hemorrhgicum in Recently Ovulated Cat 70
4.20 The Obvious Enlargement of Oviduct and Uterus in Recently 70
Ovulated Queen
4.21 The Pregnancye in Different Seasons 71

4.22 The Percentage in Different Months 71
4.23 The Migration of 2 Embryos From Right to Left Uterine Horn 73
4.24 Appearance of Reproductive System in Pseudopregnancy 74
4.25 The Vaginal Cytology During The Diestrus Phase 75
4.26 Appearance of Involuted Uterus 75
4.27 The Inactive C.L. of Involuted Reproductive System Compare 76
With Active C.L
4.28 The Mean of The Anestrus Phase According to The Seasons 77
4.29 The Anestus Phase According to The Months 77
4.30 The Reproductive System During The Anestrus 78
4.31 The Reproductive System During The Anestrus 79
4.32 vaginal cytology during anestrus phase 79
4.33 The Parovarian Cyst 80
4.34 The Parovarian Cyst in The Body 81
4.35 the Follicular Cysts 82
4.37 The Appearance of Uterus With Suppurative Endometritis 84
4.38 The Marked Thickness of The Endometrium 84
4.39 The Histological Appearance of Uterus With Suppurative 85
Endometritis
4.40 The Histological Appearance of Uterine Tube With Suppurative 86
Endometritis
Chapter one
Introduction
Introduction
Domestic cat (Felis catus ) is one of seven species of the genus Felis (a
group of small size cats),which is apart of Felids ,the family of mammals
that share a common ancestor 10-15 millions year ago (Driscoll et al., 2009
),and include in addition to domestic cat, lion, tiger, cougar and many others
(Wilson, and Reeder ,2005 ).
In past, the cats are used as familiar symbols of independence, power,

graceful, elegance and soft luxury, but now, the urban families around the
world show increasable interest to rearing a cat as household pet animal,
thus many organizations and cat fancier associations are developed to
educate people about it ( Wastlhuber ,2001).
Scientifically, domestic cats are used as laboratory animals due to the fact
that they have some physiological features more in common with human
than the laboratory rabbit or other rodent, hence they have extremely used in
behavioral and biomedical researches particularly in neurological signs
(James ,1995).
Recently, the domestic cat (Felis catus) is a useful model for studying
comparative biology of the Felidae family and for developing assisted
reproductive techniques for propagating related, endangered species (Wildt
,1991), either by embryo transfer technique or stem cell cloning (Pope et
al.,2004).
For all above, and due to the rarity of the available references about the
feline reproductive system particularly in Iraq, the present study is designed
Aims of Study
1. Study of topography and measurements of queen's non pregnant

pathological changes free reproductive systems during different
ages and seasons .
2. Study of feline follicular phase incidence, ovarian and uterine
changes, hormonal levels, and vaginal cytology during different
seasons.
3. Study of feline luteal phase incidence, ovarian and uterine changes,
hormonal levels, and vaginal cytology during different seasons
4. Recording of implantation rate and trans uterine migration in Iraqi
feral cats.
5. Study of feline anestrous phase incidence, ovarian and uterine
changes , hormonal levels ,and vaginal cytology during different
seasons
6. Study of some pathological changes of reproductive systems, which
observed during the study period, include :
 Pathological gross examination of lesions.
 Histopathological examination of lesions.
 Isolation of pathogenic aerobic bacteria involving with the
lesions.
Chapter two
Review of literature
2. Review of Literatures
2.1. The Cats
The cat is a small carnivorous mammal that is valued by mans for its
companionship and its ability to hunt household pests, and is currently the
most popular pet in the world, and cat is known to hunt over 1000 species
for food . (Siegal ,2001).
2.1.1.Scientific Classification of The Domestic Cats
The Scientific classification of domestic cat mentioned in table 2.1

according to Driscoll, et al., (2007).
Table (2.1) reveal the classification of the domestic cat
Kingdom Animalia
Sub kingdom Eumetazoa

Phylum Chordate
Sub phylum Vertebrata
Class Mammalia
Sub class Theria
Order Carnivora
Suborder Feliformia
Family Felidae
Sub family Felinae
Genus Felis
Species F. catus
2.1.2.Domestication and History
Until recently, the cat was commonly believed to have been domesticated
ancient Egypt were it was a cult animal, however, in 2004 a domesticated cat
that was buried 9500 years ago was discovered in Cyprus. Driscoll et
al.,(2009) found that lines of decent of all housed cats probably run through
as few as five self domesticating African wild cats (Felis lybica) 8000 BC
in the near east. Several countries after Christ begin to import the cat, India
was the first to introduce them in to religion and Hindu rites,in China,cats
were popular pets since the beginning of the Christian era and were
considered bearer of good fortune , in Japan domestic cats were introduced
in sixth century AD at the same time Buddhism (Vigne et al., 2004) .
The tradition of bringing cats on board ship was a result of the believe that
they brought good luck as well as to their ability as rats catcher so the ocean
did not hinder the spread of cats as they did the migration of other land
animal. Cats were able to enter the new world and Australia because of their
compatibility with ships and human explorer (Wastlhuber ,2001).
2.1.3. Hybridation with Wild Cats
Domestic cats can produce fertile offspring after mates with several of
small wild cats specially African wild cats (Felis lybica), European wild cats
(Felis sylvistris ) and jungle cats(Felis chaus) ( Wiseman et al., 2000),all
these moderate size cats are found in Iraq (Hatt,1959). Pope et al., (2004)
were succeeded in the use of the African wild cats (Felis lybica ) as a
somatic cells donor to evaluate the in vivo developmental competence after
transfer into domestic cats recipients.
2.1.4. Breeds
The famous cat breeds which registered with the Cat Fanciers
Association in the united state according to Wastlhuber , (2001) are
mentioned in table 2.2.
Table (2.2) reveals the Domestic cat's breeds
Breed Country Origin Coat
Abyssinian Egypt Natural short
American Curl United States Mutation Short/long
American Wirehair United States Mutation Rex
Burmese Burma Natural Short
California Spangled California Crossbreed Short
Devon Rex England Mutation Rex
Egyptian Mau Egypt Natural Short
Javanese - Crossbreed Long
Manx Isle of Man Natural/mutation Short/long
Persian Iran - Long
Peterbald Russia Cross breed Hairless
Russian Blue Russia Natural Short
Siamese Thailand Natural Short
Somali Somalia Mutation Long
Turkish Angora Turkey Natural Semi long

2.2. Feline Reproduction
2.2.1. Anatomy of Female Genital Tract
2.2.1.2 Ovaries
2.2.1.2.1. Morphology and Location
The ovary of adult queen is oval in shape with slightly compressed

bilaterally ,it has a smooth surface at pre puberty age, then gradually be
more granular due to present of small follicles (Johnston et al., 2001).The
ovary size and weight are vary according to the breed, age and sexual
activity of queen, in young or sexual inactive queen the ovary is measured
0.3 cm long × 0.2 cm width with proximal weight 0.3 gm ,while in adult
sexual active queen it reach 1 cm long ×0.5 cm width and about 1 gm
weight (Shille et al., 1983; Bliley &Al-Qara’awy ,2005).
As contrast to most large animal ,the feline ovaries are located in

abdominal cavity ,exactly in the dorsal abdomen caudal to kidneys (Johnston
et al., 2001).Each ovary is attached to diaphragm by suspensory ligament
,to dorsal body wall by mesovarian and to the end of uterine horn by short
,thick proper ligament. The ovarian bursa is formed by a fold of peritoneum
originating from lateral side of mesovarian which pass around the ovary,
this bursa has small slit –like opening on medial side which join between the
bursal and peritoneal cavities ( Fletcher ,1974).
2.2.1.2.2. Blood Supply
The ovarian artery is originating from the aorta ,it supplies the ovary and
caudal portion of uterine tubes ,while there are three ovarian veins drains the
ovary ,uterine tubes and cranial portion of uterine horns ,which terminate in
the caudal vena cava(Del Campo and Ginther ,1974).
2.2.1.2.3. Ovarian Follicles
The ovarian follicles is a small simple structure appear in spherical shape

ranged from 1.5-4 mm in diameter, During the ovulatory cycle the follicle
diameter progressively increases during the estrus period to reaching the
diameter of 3.2 mm in average with at least one follicle per queen being
larger than 3 mm and the maximal follicular size is reached between the 2 nd
and the 6th day of estrus with average of 4.25 ± 1.5 days. Fontbonne and
Malandain ,(2006) and Wildt and Seager , (1978) were reviewed that the
follicular phase approaches three to seven follicles enlarge ,and the rest
undergo atrasia and the most follicular development occur in the 48 hours
interval just prior to onset of estrous behavior.
2.2.1.2.4. Corpous Luteum
Corpora lutea which form after ovulation appear orange –yellow grossly
and may reach peaking in size (4.5 mm) about 13 days after ovulation (Wildt
&Seager, 1980; Wildt et al., 1981) .The formation of corpus luteum involves
luteinization of the granulosa, by which the granulosa is converted from
estrogen secretion to progesterone secretion, the process is initiated by the
pro-ovulatory LH surge (Reece, 1997).
The cavity of ruptured follicle and the fibrin clot within serve as the
framework on which the granulosa cells develop, the vessels from the theca
externa invade the developing corpus luteum, so that it becomes
vascularized maintenance of corpus luteum is provided by LH derived from
LH surge and by the basal circulating levels of LH (Reece, 1997; Bliley
&Al-Qara’awy 2005).
2.2.1.3 Uterine Tubes (oviducts)

2.2.1.3.1 Morphology and Location
The uterine tube of queen has a several parts ,the cranial end ,the
infundibulum (funnel) is located over the ovary but does not surround it as in
the bitch, the abdominal opening of funnel has fringe process called fimbrae
which erected during ovulation as a response to the effect of the oxytocin
(Chatdarong et al., 2005),the second portion is ampulla which is relatively
wide part located between the funnel and the third narrow portion which
called the isthmus that opened to uterine horn through uterine orifice((Perez
et al., 1999) .Uterine tube of queen lie craniomedial to the ovary ,from there
the uterine tube pass cranially ,laterally and caudally in the mesosalpinx
before joining the uterine horn through low papilla with a collar of circular
smooth muscle that protrude into the lumen of uterine tube ,the change in the
diameter of uterine orifice is occur under effect of estrogen and
progesterone (Johnston et al., 2001) .In adult queen the length of the uterine
tube ranged about 5-6 cm ( Fletcher ,1974).
2.2.1.4. Uterus and Cervix

2.2.1.4.1 Morphology and Location
The normal uterus of adult queen is a “Y” shape organ ,has two
relatively long uterine horns, uterine body and cervix, the short uterine body
measured 2 cm long ,lie between the descending colon dorsally and the
urinary bladder ventrally ,the horns are extended cranially to join with
uterine tubes(Spector ,1956). Cervix is the thick walled neck of uterus, it
has an internal orifice, central canal and external orifice that is opened in
vaginal end where the cervix extend a short distance into vagina (Busch
,1962).
The uterine body and horns are attached to the body wall by
mesometrium ,which is continuous cranially with mesovarian and which
with the mesosalpinx comprises the broad ligament ,the round ligament of
uterus is a lateral extension of the broad ligament passes from the horn
caudolaterally to attach near the inguinal ring (Johnston et al., 2001).
The size of uterus depends on size ,age ,parity of cat and the phase of
estrous cycle or stage of pregnancy (Reece ,1997), Bliley &Al-Qara’awy
,(2005) were reviewed the measurements of uterus parts as fallowing, in
non pregnant queen the uterus weight about 1.5 gm ,the uterine body length
is 2cm ,the uterine horn length is 10 cm and the cervix length ×diameter is
about 0.5 ×1.5cm.
2.2.1.4.2. Blood Supply

The major blood supply of the uterus is uterine artery (formerly called the
middle uterine artery) the cranial part of uterus is also supplied with blood
from the ovarian artery ,and the caudal part of the uterus receives blood from
the vaginal artery , the blood drainage of uterus occur via 2 uterine vein , the
left and right uterine vein are anostomized to the area of the uterus and
cervix (Del Campo and Ginther ,1974).During pregnancy the blood supply
to the uterus increases dramatically ,when the uterine artery is palpated ,a
vibration of the blood within it can be felt (Reece ,1997).
2.2.1.5. Vagina and Vestibule

The vagina of adult queen extend caudally from the cervix to the region
of hymen , just cranial to the external urethral orifice in the vestibule or
urogenital sinus (Fletcher , 1974) the vestibule diameter is 4mm with
about1-2 cm long , while the vagina is non distensible with about 1 mm
diameter and 1-2 cm in long , so that the distance from vulva to the cervix is
about 2-3 cm long (Zambelli &Cunto,2005)
2.2.1.5. 1.Blood Supply

The vagina and vestibule receive its blood supply from the vaginal
artery(Reece ,1997) .
2.2.1.6. Vulva and Clitoris

The vulva of the adult queen is the caudal portion of the female genitalia
that extend from the vagina to the exterior (Reece, 1997). It consist of two
small, round , lateral labia located just below the anus which unite at dorsal
and ventral commisures , during estrus the labia are slightly edematous and
reddened (Johnston et al., 2001 ).
Clitoris is consist of paired of crura of erectile tissue (the corpora

cavernousum clitoridis) originating on the ischial arch, that unite to form
body which end ,distally the erectile glans (Fletcher ,1974).The clitoris is
located in the floor of the fossa clitoridis on the ventral midline of the
vestibule ,the clitoris is homologous to the penis of male ,in the queen it may
contain a small cartilage (Bloom , 1954). Clitorial hypertrophy may occur in
response to exposure to exogenous or endogenous testosterone and may be
present in individuals with abnormality of sexual differentiation(Johnston et
al., 2001).
2.2.2. Physiology of Female Reproduction
2.2.2.1. Breeding Season
The queen is described as a seasonally poly estrous and long day
induce ovulator ,she cycle repeatedly during breeding season unless
interrupted by pregnancy ,pseudopregnancy or illness, and it’s possible to
induce estrous by social stimuli , either by a tomcat or anestrous
female(Root et al., 1995; Johnston et al., 2001) .
Estrous cycles will occur at interval ranging from 4 to 30 days, but are
typically 14-21 days apart(Wildt et al., 1918; Root,1995),and under the
natural conditions the free ranging domestic cats can produce occasionally 3
litters /year, with 1-5 kitten in each litter (pope,2000), and this is marked in
contrast to the canine where bitch may cycle only twice per year(Feldman
and Nelson ,2004; Simpson et al., 2004).In general the breeding season in
cats is determined by number of factors including the length of day light
,environmental temperature ,breed and presence of other cats(Eldredge et al.,
2008 ).
Many authors suggested that the estrous in free ranging queen is induced
by an increase in day length ,where as decreasing photoperiod results in
seasonal anestrous(Concannon and Lein,1983; Feldman and Nelson,1996 ;
Chatdarong,2003 ),and the feline estrus cycle occur at 4-30 day interval in
queen exposed to constant day length with about 14 hours bright light
period(Michel,1993).There are different data about relationship between the
breeding season and geographical area ,Eldredge et al., (2008) revealed that
the breeding season of cats in northern hemisphere is from March to
September and from October to March in southern hemisphere
.Lofstedt,(1982) reviewed various surveys document greatest frequency of
cycling activity in queen in the northern hemisphere in January and February
with gradual frequency decline until September and October ,while Tsutsui
et al., (1989) and Hurni,(1981) were reported that cat breed throughout the
year in equatorial zones and the breeding season decrease with increasing
the latitude from equator to about 6months at polar circles.
The artificial light can alter the normal ovarian activity in queens (Shille
and Sojka ,1995).Cline et al., (1980) and Randall ,(1975) reported that
queens housed indoor with constant temperature but seasonal light
(windows) cycle seasonally ,and queens housed indoor for 12 month with 14
hours of light daily show a decrease in estrous behavior only in April and
May .Inadequate intensity or duration of light is the major reason for
prolonged anestrous in queens kept in apartments and indoor
catteries(Feldman and Nelson,1996),but the effect of light on estrous cycle
of housed cat can be quiet complicated ,pet cat usually don’t receive a
consistent light pattern because of their exposure to both natural and
artificial sources ,the artificial lighting in the home may not always results in
predictable ovarian cycle ,also subjectively ,most of these queen don’t cycle
in October ,November and December (Shille and Sojka,1995).
In the temperate zones, where the queen cycle around year, the high
ambient temperature during summer may reduce the incidence of estrous
(Feldman and Nelson,1996). It has been suggested that the interestrous
period may lengthen during rather warm temperature (Concannon et al
,1989),and the effect of heat and /or humidity on ovarian function remain
subjective and depend to some degree on individual variation(Banks ,1986;
Shille and Sojka ,1995). The ovarian inactivity in Winter may be due to
elevation of Prolactin and melatonin in circulating plasma (Leyva et al.,
1984; Leyva et al., 1989).
According to the breed, long haired cat show more marked seasonal
distribution of births than do short haired cats (Johnstone ,1987),on other
hand, Banks ,(1986) record that the long hair breed is more sensitive to
photoperiod than short hair breed, and about 90% and 40% showing Winter
anestrous respectively.
2.2.2.2. Puberty
Puberty refers to the age where the animal ( male or female ) can produce
the gametes and this followed by the estrous and ovulation in the female
.The optimal breeding age of cats is between 1.5 and 7years , female older
than 7 to 8 years tend to cycle irregularly , have smaller litters , and have
more problem with abortion and congenital defects(Griffin ,2001a).
Young cat less than 1 year of age may also have irregular cycles and be
less predictable in their sexual behavior (Feldman & Nelson,2004).Many
authors record that many factors effects on the age of the breeding , include
breed ,many short hair breeds reach puberty earlier than long hair breed , the
time of year which determine the length of day light , and the body condition
of the queen (Johnston et al., 2001; Long,2006 ;Eldredge et al., 2008) .
Long ,(2006) was reported that the first estrous may occur in queens at
any time between 4 and 12 months of age , while others observed that the
puberty is beginning from 6 to 9 months of age (Reece, 1997) .
Different studies record the effect of the animal weight on the onset of the
puberty, the domestic cat is generally thought to reach sexual puberty after
she attains at least 80% of adult body weight 2.3 to 3.2 kg (Feldman &
Nelson , 2004 ) .Simpson et al., (2004) said that the cats usually start to
show estrous when they have reached about 2.3 to 2.5 kg body weight .
2.2.2.3.Estrous Cycle
The estrous cycle defined as a co-ordinate sequences of ovarian ,uterine

,vaginal and behavioral changes ,which had occur in mammals to ensure
production and fertilization of female gametes (Simpson et al., 2004).In
general, the estrous cycle of queen consist of four separate major phases
include ,proestrus ,estrous ,diestrus and anestrous (Long ,2006).However ,
the combination of seasonally polyestrous cycle and induced ovulation
makes the feline estrous cycle unlike that of bitch , also the normal queen
may undergo to a fifth phase called post estrus or interestrus (Feldman &
Nelson , 2004).
The female cat cycling repeatedly every 14-21 days(Little ,2001),but in

absence of mating and/or ovulation ,heat period are observed every 10-20
days during the breeding season(Simpson et al., 2004) .Figure 1.2 explain
the relationships between estrous cycle phases according to (Johnston et
al.,2001).
Proestrus 0-2days
Estrus≈2-19days
Postestrus Diestrus Pseudopregnant Diestrus Pregnant

≈40 days ≈60dayes
8-10 days
Anestrus ≈30-90 days
Figure (2.1) reveal the estrus cycle of domestic queen
2.2.2.4 Phases of Estrous Cycle

2.2.2.4.1. Proestrus
2.2.2.4.1.1 Definition
The Proestrus is defined as the period when the males are attracted by
non-receptive female(Simpson et al., 2004),in fact it is observed in a
minority of queens(Fontbonne and Malandain ,2006).Eldredge et al., (2008)
were described this phase as a period of courtship during which exposure to
the male act as hormonal stimulus that bring full heat, this belief stems from
the observation that in feral cat colonies where male companionship is
common, the conception rate is higher than in catteries ,where courtship is
limited.
2.2.2.4.1.2. Duration and Signs
Proestrus is not observed regularly in all cycling queen .Shille and Sojka
,(1995) were recognized this phase in27 out of 168 cycles ,and due to it
difficult to recognized, proestrus and estrous are often taken together and
just called estrous(Long,2006).However ,Feldman and Nelson ,(2004) were
observed that the queen in this phase may be less sexually demonstrative
than is noted in subsequent estrous ,she does show estrous behavior but she
doesn’t allow the tomcat to mount .Chatdarong ,(2003) was reported that
the range of duration of proestrus is 1-2 days, while Shille and Sojka ,(1995)
were recorded this duration about 0.5-2 days .
The main behavioral signs which associated with the onset of proestrus
include continuous rubbing of head and neck against any convenient objects,
constant vocalizing, lordosis posturing and rolling (Little,2001).
2.2.2.4.1.3. Hormonal Changes
During proestrus there is abrupt rise in circulating estrogen which secreted

from granulosa cells of ovarian follicles, rather than previous phase
anoestrus or interestrus (Shille, 1979), this elevation in estrogen
concentration is associated with rapid follicular growth. Although, a two-
fold increases which occur in plasma estrogen concentration to reach value
above 40 pg/ml, this concentration is considers as low or insufficient to
allow maximum behavioral expressions of heat (Feldman & Nelson, 2004).
2.2.2.4.1.4. Vaginal Cytology
The effect of estrogen on vaginal epithelium of queen it cause an

increased number of epithelial cells layer and to cause vaginal cornification
resulting in morphologic appearance of exfoliated epithelial cells
(Fontbonne & Malandain , 2006).
The vaginal cytology smears of the queen in this phase has been described
by Feldman & Nelson ,( 2004) as it consist of parabasal cells
18%,intermediate cells 60%,nucleated superficial cells 20% and anucleated
superficial cells 20%,,with about 8% neutrophils.
2.2.2.4.2. Estrus (Follicular Phase)
The queen’s estrus is defined as behavioral stage of receptivity to

mating (Little, 2001), or it’s a period of breeding, therefore this phase is
recognized solely by queen’s behavioral response to the tomcat, its begin
when she allow the male to mount and end when this behavior
cease(Feldman & Nelson , 2004).
2.2.2.4.2.2 Duration and Signs
There are various data about the duration of estrus in queen, Chatdarong
,(2003) was recorded that the estrus phase duration is ranged from 3 to 16
days with average 7days,Shille et al., (1979) were demonstrated a wide
range of duration and suggested that the healthy fertile queen may exhibit
estrus for as little as 1day to as long as21 days, and the queens experiencing
coitus are in estrus for approximately 8.5 days, but the queen that don’t
have coital contact are in estrus for 6days on average. In other study ,Shille
and Stabenfeldt ,(1979) were recorded that the estrus duration was similar in
queen that experienced coitus and ovulation (x=8.6;SD=4.1 ;n=32) and
queen that had coital contact but failed to ovulate (x=8.3; SD= 4.3;n=43)
,while Shille et al., (1979)were found that the estrus was shorter in queens
which did not have coital contact(x=6.2days ;SD=2.9 ;n=98) and the
behavior without regard for coitus in 168 cycles was 2-19days.Wildt et
al.,(1981) study the relationship between the duration of estrus and number
of follicle, and reported that were similar in ovulating (5.2±2days ;5
follicles) and non ovulating (6.4±0.6 days ;5.2 follicles) queens.
The behavioral signs of estrus include crouching with the fore quarters
pressed to the ground, rolling and hyperextension of back causing lordosis
and presentation of the vulva for mating (Stover &Sokolowski, 1978; Shille
et al., 1979; Voith, 1980). The estrous queens is frequently vocalized and
show restlessness and affectionate head rubbing to owner, as well as other
object such as pieces of furniture (Lein et al.,1982), they also crawl with
their thorax against the floor and she will raise her hindquarter (Reece
,1997;Eldredge et al. ,2008).
2.2.2.4.2.3.Horminal Changes
As the estrus phase is described as the phase of follicular activity it could

be recognized biochemically via plasma estrogen concentration ,the
evidence of follicular activity is defined as plasma estrogen concentration
reaching a high of 20 to 50 pg/ml (or higher) ( Long,2006), Feldman and
Nelson ,(2004) were detailed the hormonal changes in different estrous days
as following :during the follicular phase ,the plasma estrogen concentration
rapidly increases and remain elevated for 3-4 days, and abruptly begin to
decline. The first day of follicular phase is associated with estrogen
concentration of approximately 25 pg/ml and rising to approximately 45
pg/ml on day 3,slightly above 50 pg/ml on day 5 and from 12-25 pg/ml on
day 7 and usually back to 10 pg/ml on day 8.Shille and Stabnfeldt,(1979)
were reported that the peak of plasma estradiol may reach 70 pg/ml ,while
Griffin ,(2001b) was explained that the raised level of estrogen are necessary
to induce estrogen behavior and prime the anterior pituitary gland for the
luteinizing hormone(LH) surge that cause ovulation .Simpson et al. ,(2004)
were suggested that after estradiol-17 ß rise ,if the no ovulation occur it will
decrease to basal concentration within 5-10 days and if ovulation occur the
concentration will decline within 2-3 days.
2.2.2.4.2.4.Vaginal Cytology
Johnston et al.,(2001) and Shille et al.,(1979) detailed the vaginal

cytological changes during estrous phase as fallowing :in estrous phase ,the
anuclear superficial cell will increase to 10% of the total on the first day of
follicular phase and continue to increase in number to reaching
approximately 40% of total from the 4th to 7th days, in the same time the
intermediate cells decrease in number from approximately 40-50% in
interestrous period to less than 10% during the first 4 days of follicular
phase, the number of superficial cells is fairly constant throughout behavior
estrous ranged from 40-60 of cells present .Erythrocytes and leukocytes are
rarely seen in the feline vaginal smear during proestrous or estrous and
parabasal cells comprise less than 10% of the total cells at all stages of the
cycle(Shille et al.,1979).
2.2.2.4.3. Interestrus
2.2.2.4.3.1.Definition
The interestrus phase defined as the brief period of sexual or reproductive
inactivity which separate between sexual activity waves of follicular
function(Feldman and Nelson,2004),also it called interestrus interval or post
estrus (Lofstedt, 1982).
2.2.2.4.3.2.Duration and Signs

This phase is observed between two periods of intense sexual activity,
during which the cat show no specific physical or behavioral signs .The
duration of this phase is probability not affected by coital contact that fail to
induce ovulation (Feldman and Nelson, 2004).The interestrus period in
queen experiencing coitus,but not ovulate were shorter in duration than
those in animals that had not experience coitus(8days versus 10 days,
respectively)(Shille et al., 1979) . Chatdarong,(2003) was suggested that if
the queen is not mated ,ovulation will not occur and the pro ovulatory
follicles become atrectic and degenerated ,but the interestrus period is much
longer than 8 days in average if ovulation is induced(Pseudopregnancy and
pregnancy).The ovulation result in corpus luteum development that secret
progesterone (Johnston et al.,2001).There are different interestrus durations
which recorded by many authors ,include 2-10 days(Lofstedt,1982),2-12
(Shille and Sojka,1995) and 14 days (Long,2006).
The interestrus period is characterized by a return to normal personality

and the queens don’t breed, don’t attract males and lose the extremely
affectionate behavior pattern that typify estrous, the intense rubbing ,rolling
and vocalizing are disappear(Johnston et al., 2001; Chatdarong,2003) .
The interestrus period follow cessation of follicular function ,which is

characterized by an abrupt decline of plasma estrogen concentration to level
below 20 pg/ml (Wildt and Seager ,1980 ;Shille&Sojka,1995;Johnston et
al.,2001),and this concentration remains at the low or basal level throughout
this phase to become just below 20 pg/ml during proestrus (Chatdarong,
2003 ;Long,2006).
2.2.2.4.3.4. Vaginal Cytology
Mills et al.,(1979) described the cells comprising the vaginal smear in

queens during this phase as present in table 2.3
Table (2.3) reveals the vaginal cytology in interestrus phase of queen
Cell type Range % Mean %
Para basal cell 0-29 8.9
Intermediate cells 50-100 75
Nucleated superficial cells 0-14 13.2
a nucleated superficial cells 0-3 1.4
Neutrophils 32/100 epithelial cells 1-78
2.2.2.4.4. Anestrus
The anoestrus is defined as the period of clinical reproductive quiescence
and a seasonal absence of cycling activity (Feldman &Nelson,2004)
.Long,(2006) detailed that the anoestrus is the period during the winter when
there is no ovarian activity.
2.2.2.4.4.2 Duration and Signs
Anoestrus occurs when animals are living under very short natural day
light (winter)(Simpson et al.,2004) . It occurs in late autumn months include
October ,November and December in queens exposed to short light interval
of 4-6 h/day in the natural photoperiod in the northern hemisphere ,but may
be absent in animals submitted to constant long light day (Johnston et
al.,2001; Simpson et al.,2004).
Anoestrus typically has been described in some colonies and / or

individual queens as beginning in mid summer (June through august) and
lasting through January,and the heat of the summer is the other factor that
may induce the onset of anoestrus(Concannon & Lein ,1983).The female in
anoestrus dose not revealed any attracting to the male and does not display
sexual behavior or exhibit evidence of active ovarian function . It is difficult
to differentiate the queen in anoestrus from one that has been
ovariohysterectomized(Little,2001).
Hormonally, the anoestrus period is due to the secretion of melatonin

from the pineal gland ,melatonin is secreted during the hours of darkness and
suppresses the release of gonadotropin releasing hormone(GnRH) from the
hypothalamus ,thus the anterior pituitary gland does not release the
gonadotropins which are the follicular stimulating hormone (FSH) and
luteinizing hormone(LH) that would stimulate the ovaries (Noaks et
al.,2003),this will remain the concentration of the estrogen and progesterone
basal levels( Johnston et al. ,2001;Simpson et al.,2004) . Banks and
Stabenfeldt,(1983) were recorded Serum prolactin concentrations in six
anoestrus queens during a7 weeks period of 7 hours of light per day
averaged 13.2 ± 0.5 ng/ ml .
The vaginal cytology of the anoestrus is consistent with an interestrus

period, during anoestrus ,cells are rare , mucus is obvious and clear ,and the
majority of cells are basal or parabasal with some intermediate cells
leukocyte can sometimes be observed(Simpson et al.,2004). Mills et
al.,(1979) were describe that the exfoliated vaginal epithelial cells consist of
less than 10% parabasal cells,40%to70% intermediate cells , and 30% to
40% nucleated superficial cells ,background debris is obvious . and different
data was recorded by Johnston et al. ,(2001) which are present in table 2.4.
Table (2.4) reveals the vaginal cytology in anestrus phase of queen
Cell type Range % Main percentage
Para basal cell 0-50% 9.7%
Intermediate cells 50-100% 87.4
Nucleated superficial cells 0-22% 2.7%
a nucleated superficial cells 0-2% 0.2%
Neutropills 0 – 50 3/100epithelialcells
2.2.2.4.5.Dioestrus
2.2.2.4.5.1.Definition
Diestrus defined as a phase of progesterone dominance in the queen
(Johnston et al., 2001), she must have coital contact or similar vaginal
stimulation to induce ovulation before corpora lutea develop (Feldman
&Nelson, 2004).If ovulation occur, but she does not become pregnant,
diestrus will follow (Griffin, 2001b) .
2.2.2.4.5.2. Duration & Clinical Signs
Corpora lutea will form in this phase within 24 -48 hours of ovulation and
begin secreting progesterone (Feldman and Nelson, 1996).They remain
functional for 30-50 days (average35 days) in non pregnant queens ,at which
time regression occurs ,an interestrus interval follows (Griffin,2001b).
Schmidt et al.,(1983) detailed that the diestrus last approximately 40 days in
the Pseudopregnancy queen and approximately 60 days in the pregnant
queen, but Feldman&Nelson,(2004)were said that the ovaries contain
several firm , tan to orange corpora lutea that remain functional for 35 to 73
days .The uterus is largest in diestrus , because of the extreme thickening of
the endometrium ,the horn become turgid and twisted inside the serosa
,forming irregular corkscrew bulges that may be palpable ,vagina, vestibule
and vulva are normal in appearance ,on rare occasions , queens exhibit
behavioral signs of estrus at the termination of diestrus(Feldman &
Nelson,2004).
2.2.2.4.5.3.Hormonal Changes
Johnston et al.(2001) were detailed that the serum progesterone

concentrations range from 1,5ng/ml to more than 20 ng/ml in the diestrus
queens ,it ends with luteolysis , when serum progesterone concentration fall
below 1.5 ng/ml( Schmidt et al.,1983) . progesterone concentrations decline
to basal levels , in such cases ,hormonal estrus is absent and estradiol
concentrations remain at basal levels (Griffin ,2001b).plasma progesterone
concentrations greater than 1-2 ng/ml are associated with diestrus and are
greater than 20 ng /ml within14 to18 days.
Tsutsui&Stabenfeldt, (1993) were described the cells comprising the

vaginal smear in eight late diestrus cats as in table 2.5.
Table (2.5) reveals the vaginal cytology in Dioestrus phase of queen
Cell type Range % Main percentage
Para basal cell 10-85 48%
Intermediate cells 10-85 50%
Nucleated superficial cells 0 -5 2%
a nucleated superficial cells - 0%
Neutropills 0 -97 32/100epithelialcells
2.2.2.5. Mating and Ovulation
The cat is considered as a reflex mediated induced ovulator (Johnston et

al.,2001; Griffin,2001a; Simpson et al.,2004; Michael et al.,2008) , that
mean the coitus is required for ovulation (Reece,1997) .
The ovulation rate is vary according to breed , number and timing of

mating (Simpson et al.,2004).Long,(2006) was documented that the
ovulation is easier in some breed than others such as , the oriental breeds
require fewer mating to stimulate ovulation than short hair breeds . In
oriental breeds and in some circumstances the ovulation will occur without
mating by stroking the back (Gudermuth et al.,1997), furthermore ,cat kept
in colonies may ovulate without being mated , this phenomenon may be due
to the phermonal activity within the colony or the sexual activity of other
queens (Long ,2006) , so the spontaneous ovulation seen to occur more
frequently in old cats (Simpson et al.,2004) .Chatdarong,(2003) suggested
that the mechanism of induced ovulation as a coital stimulus will induces a
neural firing reflex that stimulates the medial basal hypothalamus to
synthesis and libration gonadotropin releasing hormone (GnRH ) , this will
stimulate the release of luteinizing hormone (LH) (Robinson
&sawyer,1987). Long, (2006) was reported that the many successive mating
are important to trigger ovulation and release of GnRH which trigger LH
release from anterior pituitary gland . The luteinizing hormone is released
within a minutes following coitus or after administration of exogenous LH
releasing hormone (LHRH) (Chakraborty et al., 1979; Fontebonne
&Malandain, 2006).
The magnitude of LH release has been demonstrated to increase with

increase in number of mating , and only about half the queens bred a single
time will secrete adequate LH to induce ovulation (Johnston et al.,2001) , in
general , the maximal LH level are attained 4 hours after the onset of 8-12
copulations , while some queens are require at least 4 copulations to release
enough LH (Eldredge et al.,2008) , the interval between coital stimulation
and ovulation has been reported to vary in cats ,from 25 -50 hours (Shille et
al. , 1983).
2.2.2.6.Pseudopregnancy
2.2.2.6.1.Defenition
It is a physiologic alteration that usually occurs when queens has been
induced to ovulate but fails to become pregnant , or if the ovulated oocytes
are not fertilized (Chatdarong ,2003),while Michael et al.,(2008) defined the
pseudopregnancy as a condition in which the female display the physical and
emotional signs of pregnancy without carrying the kittens . The queens may
undergo as many as 4-5 pseudopregnancies during the course of one poly
estrus season (Feldman & Nelson, 2004).
2.2.2.6.2. Etiology
There are many natural and artificial causes of pseudopregnancy ,which

lead to induce ovulation without subsequent fertilization , these include ,
male infertility , inhibiting contact between sperms and ova ,artificial
induction of ovulate (mechanical or hormonal ) genital tract problems
(Chatdarong ,2003).
2.2.2.6.3. Signs and Duration
The obvious signs of pseudopregnancy are very rare in cats

(Griffin,2001a) but when present , it were mild , include lactation , nesting
and tending kittens (Feldman &Nelson,1996) , the queen’s abdomen may be
distended with increase of appetite and weight gains , and sometimes it may
exhibit constant meowing (Griffin,2001b;Mechael et al.,2008).
Concannon& Lein,(1983) were revealed that the average of duration of

luteal activity in pseudopregnancy was 36.5 days , with range of 30-50 days
, presence of corpora lutea in case of pseudopregnancy ranged 30-70 days ,
while Feldman& Nelson,(2004)were reported that the actual luteal phase
lasts about 36-73 days , followed by 7-10 days interestrus interval.
2.2.2.6.4. Hormonal Changes
The basal progesterone concentration which associated with no functional

corpora lutea is <0.5 mg / dl (Feldman& Nelson,2004), the luteal activity
during pseudopregnancy is begins about 4 days after the first day of
copulation , the plasma progesterone concentration will increase above 1 mg
/ dl and reach the maximal level around 20-25 days after mating
(Verstegen,1998) , then begin to decline gradually at days 25 to reach basal
value in 30-40 days after the first day of copulation (Verhage et
al.,1976;Shille and Stabenfeldt,1979;Shille&Sojka,1995) .
2.2.2.7. Pregnancy
2.2.2.7.1.Definition
It is a physiologic alteration that occurs when queen has been induced to
ovulate and become pregnancy, in free roaming queens there is a bimodal of
pregnancy , most kittens are born in midspring and late summer
(Schmidt,1986).
2.2.2.7.2 Gestation Length

Gestation length defined as the interval from a fertile mating to
parturition ,It is variable due to the relatively long period of sexual
receptivity and the un certainly of the time of the ovulation and conception
and it may be divided in to three trimester , each approximately 3 weeks in
duration(Griffin,2001b).
The queen gestation period is 65-67 days on average with a range of 62-
71 days (Root et al., 1995), while others recorded that the length of feline
gestation is ranging from 42-75 days timed either from the first or the last
breeding (Munday&Davidson, 1993; Johnston et al. ,2001).
2.2.2.7.3. Behavior
Many queens become more docile during the first two trimester and
exhibit pinking of the nipples and the nipples become pinker and more erect,
by the first trimester ,the behavior changes are obvious which include
abdominal distention ,enlargement of mammary gland , excessive grooming
of the mammary gland and perineal area and nesting(Beaver,1992).
2.2.2.7.4.Hormonal Changes
The progesterone concentration during the initial 14 -20 days of

pregnancy is similar to those seen in pseudopregnant cats
(Feldman&Nelson,2004),thereafter, plasma progesterone concentration are
increased in pregnant queens (Concannon&Lein,1983), in addition
,progesterone begins to be synthesized and secreted by the placenta after 30
days of gestation ,plasma progesterone concentration very markedly among
pregnant cats around 5-50ng /ml (Shille&Sojka,1995).The serum
progesterone concentration are not helpful in diagnosis of pregnancy
because they do not significantly differ in pregnant and pseudopregnant
queens(Feldman&Nelson,1996) .
Relaxin is the only pregnancy specific hormone in cats(Reece,1997;

Verstegen, 1998;Griffin,2001b),prolactin is elevated in the blood around 25-
30 days and only after relaxin rising (Reece,1997 ;Feldman& Nelson,2004 )
.Klonisch et al., (1999) revealed that by gestation day 40-50,
ovarioctomydoes not result in termination of pregnancy because placental
progesterone maintains pregnancy at the end of gestation ,plasma estrogen
and prolactin concentration and relaxin which produce from the placenta.
2.2.3Pathology of Female Reproductive System

2.2.3.1.Disorders of The Ovary
2.2.3.1.1.Ovarian and Parovarian Cysts
2.2.3.1.1.1Cystic Rete Ovarii
Cystic rete ovarii is very common in domestic cats, and occur as result to
distention of rete ovarii with fluid , and it would be expected that the rete
enlarge with age , they common in young as well as old cats
(McEntee,1990).
Gelberg et al.,(1984) described cystic rete ovarii in 20 cats , the cysts were
within the ovary or at the tubal extremity of the ovary , and cyst lining was
variable , they were single or multilayered ,cuboidal to columnar ,ciliated
and non ciliated or combination of these .
2.2.3.1.2.2.Follicular Cysts
Follicular cysts that arise from mature or atretic follicles are reported to
be the most common type (Gelberg et al, 1984), and the etiology is still
unknown (Johnston et al.,2001).The normal follicles of the queens is
supposed to be up 2-3mm in diameter, the larger follicles are said to be
cystic, the affected cats show prolonged estrus or persistent estrus because a
persistent source of estrogen (Feldman and Nelson ,2004;McEntee,1990).
The follicular cysts are visible grossly and ranged from 1- 2.5cm in
diameter. Gelberg,(1984) was observed cysts as unilateral in 16 cats and
bilateral in four from 20 cats ,while Romagnoli,(2003) was reported that the
cats have both functional and non functional ovarian cysts .
2.2.3.1.2.3. Serosal Inclusion Cysts
The inclusion cysts is a cystic structure forms when there is trapping of

mesothelium from the serosa in a region beneath the serosal surface ,they
appear as a thin walled fluid filled cysts that are visible on the surface of the
ovary , (McEntee,1990)
2.2.3.1.2.4.Parovarian (Periovarian) Cysts
The cysts that occur outside the ovary are commonly called parovarian
cysts, it arises either from remnant of the mesonephric (walffian) tubules or
the paramesonephric (Mullarian) tubules (McEntee,1990).
2.2.3.1.3.Ectopic Adrenocortical Parovarian Nodules
Ectopic adrenocortical parovarian nodules have been recognized in many

species, in the cat they occur in the broad ligament of the ovary (Johnston et
al., 2001), and they occur as a single, unilateral nodules, as bilateral nodules,
or as two nodules on a single side (Altera&Miller, 1986). These nodules
ranged in size from 2- 5 mm in diameter and were smooth well delineated
firm and gold to tan in color (Johnston et al., 2001).
2.2.3.1.4. Ovarian Neoplasia
The incidence of primary ovarian neoplasia in the cat is low ,the

occurrence and behavior of ovarian tumors are influenced by the tissue of
origin within the ovary (Stein,1974), table 2.6 reveal the main ovarian
neoplasia which recorded in domestic cat.
Table (2.6) reveals some of disorders of queen’s ovary
Disorder Reference
Granulose Cell Arnbjerg,1980 Gelberg& cEntee,1985

Tumors Aliakbri&Evoghli,1978 Engle,1969
Norris et al.,1969
Interstitial Cell Kennedy et al.,

Tumors 1998
Adenoma Nielsen,(1964)
Adenocarcinoma Norris et al., (1969)
Dysgerminoma Maclchain,1987)
Teratoma Dehner et al.(1970); Basaraba et

al.,(1998) ;Sato et al. ,(2003
2.2.3.2. Disorders of the Uterine Tube
2.2.3.2.1. Hyperplasia of the Uterine Tube

It’s an occlusion of uterine tube at the uterotubal junction with
hyperplasia of oviduct villi (Cline et al., 1981). Herron, (1978) was
described a bilateral papillary hyperplasia and stromal edema of the uterine
tube in an infertile 3 years old Himalayan queens with uterotubal junction
obstruction, also has been described .
2.2.3.2.2. Salpingitis
It is inflammation of uterine tube, which usually associated with

endometritis-pyometra as an ascending infection, the agents are probably
same as those causing endometritis, but studies are lacking (Gelberg and
McEntee, 1986). The gross appearance of salpingitis varies with severity and
may cause perioophoritis as the infundibulum is affected and the tissues
around the ovary are included in the reaction (Shehata, 1974) .
2.2.3.3. Disorders of The Uterus

2.2.3.3.1. Cystic Endometrial Hyperplasia
The physiological hyperplasia of the endometrium occur during the luteal
phase of the cycle and subsequent atrophy occurs during anoestrus
(Agudelo,2004). The ovarian androgens and ovarian interstitial endocrine
cells may play a role in the occurrence of this case (Perez et al. ,1999).
Chatdarong et al. ,(2005) thought that the progesterone alone will cause
endometrial hyperplasia , and the endometrium becomes thickened with
fluid filled cystic structures .In uncontaminated case, the fluid is clear and is
present within cystic structures in the uteri mucosa , this fluid if free, in the
uterus represented the excellent medium for bacterial growth
(Feldman&Nelson,2004).
Misirlioglu et al. ,(2006) were examined 10 normal cats and 20 cats with
cystic endometrial hyperplasia and pyometra, they found that the
endometrial hyperplasia group could be divided in to 2 sub groups ,those
with mild hyperplasia and those with marked hyperplasia, while Perez et al.
,(1999) were suggested that the feral queens did not have cystic endometrial
hyperplasia . Agudelo, (2005) was documented that interesting to note that
the disorder has been described in queens over 5 years of age that have never
had kittens.
2.2.3.3.2.Endometritis and Pyometra
Endometritis is the inflammation of the endometrium, pyometra is the

uterine inflammatory disorder characterized by cystic endometrial
hyperplasia (Johnston et al., 2001). Pyometra has been observed in intact
and ovariectomized queens treated with exogenous progesterone
(Long,1972), and the age of the effected animals ranges from 1-20 years
(Kenney et al,1987,Potter et al.,1991).
The histologic features of the feline uterus with pyometra include cystic
endometrial hyperplasia, endometritis,endometrial polyps and endometrial
atrophy (Potter et al., 1991). The endometrial hyperplasia is invariably
present in cases of endometritis and pyometra (Misirlioglu et al.,2006).The
main bacterium which often isolated from fluid in the pyometric uterus of
the cats is E.coli which also is one of the most commonly isolated organisms
from the vagina of healthy cats(Kenney et al.,1987;Davidson et al.,1992).
Staphylococcus spp. ,Pastuerella spp., Klebsealla spp., Moraxella spp., and
Pseudomonas spp. also have been cultured from feline pyometric uterus
(Wilson et al.,1988) .Clemeston and Ward ,(1990) was founded bacteria in
the uterus of 2 out of 29 cats sampled .
2.2.3.3.4.Acute Metritis
It is a bacterial infection of the lining of the uterus that spreads upward

through the birth canal during the birthing process or immediately thereafter
un sanitary kittening quarters can cause it (Eldredge et al., 2008). It is seen
in 12 to 96 hours following parturition and involves infection secondary to
disorders such as retained placenta, retained fetus and injury from
instruments other trauma (Feldman & Nelson, 2004).
Table 2.7 reveal some of disorders of queen’s uterus
Disorder Reference
Endometrial Polyps Gelberg & McEntee,1984; Abel,1990; Potter et

al.(1991).
Adenomyosis Bloom ,(1954); McEntee,(1990);
Mucometra/Hydrometra Van Haaften and Taverne,1989;Abel,1990
Segmental Aplasia Memon and Schelling , 1992; Marcella et al.

,(1985)
Uterine Torsion Manda, 1986;Biller and Haibel,1987;Young et

al.,1992
Uterine Neoplasia Sorribas,1987
Leiomyoma Miller et al.,2003
Leiomyosarcoma Cooper et al.,(2006) ;Miller et al. ,(2003)
Endometrial Stromal Papparella and Roperto, (1984); Sorribas,

Tumors (1987); Sato et al. ,(2007)
Mixed Mullerian Evans and Grant, (1977); Papparella and

Tumor,MMT Roperto,(1984); Nicotina et al. ,(2002)
Haemangioma Fukui and Matsude ,(1983);

2.2.3.4.Disorders of queen’s Vagina
2.2.3.4.1. Vaginal Prolapse
Vaginal prolapse (non obstetric) is rare in the queens ,but may occur
during estrus or anoestrus (Joshua,1965).A prolapsed vagina removed from a
14 years old anoestrus ,domestic short hair cats at ovariohesterectomy had
histologic evidence of marked edema with surface ulceration and
inflammation(McEntee,1990) .
2.2.3.4.2. Vaginitis
The primary vaginitis is rare in the cats , the clinical signs include
polyuria , dysuria , frequent cleaning of the vulva , and vaginal
discharge(Johnston et al., 2001) .
2.2.3.4.3Vaginal Neoplasia
Vaginal tumors are un common in cats ,and many reports in this species
are of single occasional cases in spayed or intact older females (Whitehead,
1967 ;Stein ,1981).The most common vaginal tumor type in the cats is the
leiomyoma which may measure up to 7×7×8 cm (Wolke,1963).The clinical
signs of the vaginal tumors include bulging of the perineal region ,prolapse
of the tumor tissue from the vagina dysuria ,polyuria and
constipation(Hoover&Griesemer,1971).
Chapter three
Materials and Methods

3. Materials and Methods
3.1. Materials
3.1.1. Animals
The study was carried out in three governorates include ,Babylon, Al-
diwaniya and An-najaf from December 2008 to December 2009 , 60 mature
female cat were captured by local made feral cat trap* figure 1.3 , each
captured animal was anaesthetized according to(James ,1995) by using
Kitamine** and Xylazine*** in a dose of 10mg/kg b.w. and 0.15 mg\kg
b.w. respectively ,then the anaesthetized animal was examined clinically
with recording of obtained data in special examination card (appendix1).
Figure (3.1) the local made feral cat trap
*The feral cat trap was designed by Dr. Haider M.Al-Ramahi

** OBOI,laboratories/India
*** Interchemie/Holland
3.1.2. Sampling
3.1.2.1. Blood Samples
60 blood samples collected from cephalic vein according to McCumin

and Bassert,(2002) by 21 gauge needle fitted to 5 ml disposable syringe,
then the blood was poured in labeled plastic plain tube ,these tubes were
placed in room temperature for 2 hours , and the serum collected by
Pasteur’s pipette after centrifugation at 3000/rpm for 10 minutes, the
obtained serum was placed in labeled screw-capped plastic container which
stored in -20oC until laboratory investigation .
3.1.2.2.Vaginal Smears
The vaginal smears was employed to estimate the stage of estrous cycle
by evaluation of exfoliated cells. It was carried out according to the method
that is recommended by Fontbonne and Malandain, (2006), briefly by
introducing a cotton- tipped swab moistened with sterile saline in to the
vestibule and caudal vagina with avoiding of clitoral fossa during the
procedure because the superficial cells from this area could alter the
cytological interpretation. The cotton tipped swab was rolled onto a glass
slide, allowed to air- dry and stained with Giemsa stain.The correlation
between vaginal cytology and stages of the estrus was interpreted according
to Simpson et al.,(2004) .
3.1.2.3.Female Reproductive Tract Samples
Fifty five samples of the female reproductive tract are collected directly
after euthanizing of animal, the female reproductive tract was taken entirely
from the ovaries to the vulva for measurement and preservation.
3.1.3. Culture media
Name of media Company-origin
Nutrient agar Oxoid - England

Blood agar Mast -UK
MacConky’s agar Oxoid -England
Brain-heart infusion broth Mast -UK
Simmon’s citrate agar Biolife -USA
Peptone Difco - USA

Kliger iron agar Oxoid-England
Urease base medium Biolife - USA
Manitol salt agar Difco USA

Peptone water medium Microbiologics-
USA
Sugar fermentation medium Difco-USA
MR-VP Difco -USA
3.1.4. Reagents and Solutions

A. Reagents
Reagent Company- Origin
Oxidase reagent BDH- England
Catalase reagent BDH- England
Voges proskauer BDH- England

reagent
B. Solutions
Solution Company – Origin
Gram’s stain Crescent Diagnostics - KSA
Gimsa’s stain Crescent Diagnostics - KSA
Formaldehyde 10% BDH – England
3.1.5. Chemicals
Chemical Company – Origin
Absolute ethanol BDH - England

NaCL BDH - England
Glucose, fructose ,lactose, Difco - USA
maltose, and sucrose
H2Ο2 Fluka - Switzerland
3.1.6. The Diagnostic Equipments
The diagnostic equipment which used was Api20-E made by

Biomerieus company- France .
3.1.6. Different Materials
Name Company – Origin

Human blood O+ Al-Diwaniya teaching
hospital
Rabbit plasma Microbiologics - USA
3.2. Methods
3.2.1. Designing of Study

The study was designed as shown in figure 2.3, briefly as
following:
A. Reproductive systems were classified to pregnant and non pregnant

samples.
B. The non pregnant samples classified in to reproductive tract with or
without pathological changes.
C. Every non pregnant and has pathological changes reproductive tracts
were examined for gross pathology and histopathology as well as
bacteriological examination.
D. The reproductive tracts which appearance normally were examined
for anatomical study (topographical description as well as recording
of different measurements of reproductive tract) and Physiological
study of different phases of estrous cycle description of physiological
changes, hormonal level and vaginal smear for each phase.
E. The reproductive tracts with pregnancy were examined to record the
number of the fetuses , ovary status ,implantation rate , transtrauterine
migration of embryos and hormonal examination.
F. The age of each captured animal was estimated roughly to less and
above one year according to Van Aarde ,(1983).
06 sexually
mature queens
06 blood
06
samples
reproductive
systems
Hormonal
examination
Pregnant reproductive Non pregnant

systems (11 animal ) reproductive systems
(49 animal )
Reproductive systems Reproductive systems

 Incidence of pregnancy. not reveals reveals pathological
pathological
 Number and distribution changes(3 animal)
changes(46 animal)
of fetuses
 Implantation rate.
 transtrauterine
migration of embryos.  Anatomical  Gross description of
study(Topography pathological changes
&measurement of  Histopathological
different parts. examination
 Physiological study  Bacteriological
(estrous cycle examination
phases,hormonal
Figure (3.2) reveal the steps of study design
3.2.1.1. Non Pregnant Reproductive Systems Samples
The non pregnant reproductive tract was dissected and removed

entirely from recently euthanized animal, and examined for any abnormal
changes .
 A longitudinal incision was made along whole tract to expose the

luminal surface for examination to observe any abnormalities or
pathological changes, if reproductive systems showed clear gross
pathological lesions , small piece 1cm3 were fixed in 10% buffered
neutral formalin for 3 days for histopathological examination
according to Luna, (1968), while another piece were taken for
bacteriological examination , the bacteriological identification
performed according to Collee et al, (1996).
3.2.1.2.Pregnant Reproductive Systems Samples
For examination of these samples, the number of fetuses per animal,

number of fetuses in each uterine horn and the number of corpora lutea
were obtained.
The transuterine migration of embryos and implantation rate were

recorded according to Tsutsui et al.,(1989) as following :The number of
ovulation was estimated by number of CL in the ovaries .The number of CL
was determined by external examination of the ovary with active CL which
being round shape with a 3mm diameter and raised 2mm above surface of
the ovary,the color is pinkish with evidence of blood vessels on surface .All
structure characteristic of CL were incised and examined internally for size
(4-5mm) and color (yellow) indicative of active CL .Small whitish yellow
CL about 1-2mm in size were judged to be from the previous cycle. Each
uterine horn was opened and the number of fetuses was recorded.
Transuterine migration of embryos was estimated to have occurred prior to
implantation when the number of fetuses in a given horn was more than the
number of CL present in the ipsilateral ovary. The implantation rate
considered as 100% when the number of total CL was equal to the total
number of fetuses.
3.2.1.3. Measurement of Reproductive Tract
The measurement data were obtained from non pregnant reproductive

tract without any pathological changes as following:
A. Ovary
After dissecting and detachments of both ovaries from oviduct and

surrounded tissue , the measurement of length, width and thickness will
obtained by caliper vernier (Nichi-Japan) .
B. Oviduct
Both oviduct were dissected and detachment at uterine orifice ,the

measurement of straight length was recorded by caliper vernier.
C. Uterus
After removing of ovaries and oviducts from reproductive tract , the
length and external diameter of both uterine horn , uterine body , and
cervix will measured by the caliper vernier .
3.2.1.4. Hormonal Examination
The stored sera were send private laboratory for determination of

estradiol 17-ß and progesterone hormones by ELISA (enzyme linked
immunosorbant assay) apparatus (Bio active DNM-9602/Germany) with
specific kit ( Abbot laboratories USA) and according to the instruction of
the company .
3.2.1.5.Bacterial Isolation
The obtained swabs were send within transport media to the Al-
diwanyia hospital for bacteriological examination under supervision of
specialist staff .
3.2.1.5.1Preparation and Sterilization of Media
The above mentioned media were prepared according to the

instructions of company ,after adjusting of PH ,the media were sterilized
in autoclave at 121ΟC , 15 bar for 15 minute , while the sugars and urea
sterilized by filtration.
3.2.1.5.2. Preparation of Reagents
A. Oxidase reagent: prepares from 1% solution of Tetra- methyl –P-

phenylene diamine dehydrochloride.
B. Catalase reagent : prepared at 30 % of hydrogen peroxide
C. Voges – proskauer reagent : it consist of
1. Reagent A : 3ml of 5% α- naphthol in absolute ethyl alcohol
(Collee et al.,1996).
2. Reagent B: methyl red reagent, which reset from dissolving of 0.1 g
methyl red in 300 ml ethyl alcohol and 200 ml of distilled water

(Cruikshank et al.,1975) .
D. Kovac’s reagent: it prepared by dissolving of 10 g P-diethyl-

aminobenzyldehyde in 50 ml iso amyl alcohol, then adding of
concentrated hydrochloric acid slowly and stored at 4 ΟC until
use
(Cruikshank et al., 1975).
3.2.1.5.3. Isolation of Bacteria
Bacteriological isolation was carried out by streaking of blood agar ,Mac

Conkey agar by sampling swabs and incubate at 37oCfor 18- 24 hours .
3.2.1.5.4. Identification of Isolates
The bacterial isolates were identified according to followings:
1. Colony properties : the morphological properties of bacterial colonies

include ,the shape ,color , surface ,consistency, transparence ,hamolysis
pattern on blood agar and lactose fermentation on Mac Conkey agar
(Sneath et al.,1986 ).
2. Direct microscopical examination a direct smear from bacterial colonies
was fixed and stained with Gram’s stain to observe the shape
arrangement ,stain reaction , spore formation and present of capsule.
3. Biochemical tests :
A. Gram’s positive Bacteria
1. Catalase test:- this test carried out by mixing of small portion of bacterial
colony with 30% of hydrogen peroxide on clean glass slide , the release
of bubbles considered as positive result (Baron and Finegold,1990) .
2. Coagulase test:- after mixing of one drop of sterile normal saline with
small portion of bacterial colony on the clean sterile glass slide , add a
loop full of rabbit plasma to the mixture , the positive result is describes
as coagulation of plasma during 5-50 seconds and this reveal the
presence of coagulated enzyme. The negative result must be confirmed
by test tube to reveal the free enzyme by dilution of plasma 1:10 and
placing 0.5ml from it in to clean sterile test tube , then mixed with 0.5 ml
of bacterial suspension and incubate at 37Co , the coagulation of plasma
within 2-4 hours considered as positive result .
3. Mannitol fermentation test : this test used for differentiation of mannitol
non fermenting and fermenting Staphylococcus aureus by inoculation of
bacterial isolates on mannitol salt agar , the later were growth as yellowish
colonies .
4.
B. Gram’s Negative Bacteria :

1. Oxidase test : the oxidase reagent was added to bacterial colony on the
nutrient agar , the appearance of purple color during 10-60 second

considered as positive result (Baron and Finegold, 1990)
2. Indol test : after inoculation of bacterial isolates on peptone water ,
incubate at 37Co for 48 hours , add 0.5 ml of Kovac’s reagent with

persistent mixing , the appearance of red ring considered positive result
(Collee et al.,1996).
3. Citrate utilization test : the bacterial isolate was inoculate on slope citrate
agar and incubate at 37 Co for 24 hours ,the change of slope surface from
green to blue color considered as positive result (Cruickshank et
al.,1975).
4. Methyl red test : in this test the isolated bacteria was inoculated on MR-
VP and incubate at 37CΟ for 48 hours add 5 drops of methyl red reagent ,
the changing of media to red color due to acid production was
considered as positive result , while the changing of media to yellow
color considered as negative result (Collee et al.,1996).
5. Voges proskauer test : the isolated bacteria was cultured on MR-VP
media and incubate at37 Co for 24 hours ,add 6 drops of reagent A with 2
drops of reagent B for every tube contains the incubated media , after 15
minute ,the positive result occurred as appearance of red color , while the
negative result occurred as appearance of yellow color (Baron and
Finegold ,1990).
6. Sugar fermentation test :the isolated bacteria was inoculated on sugar
fermentation media at 37 Co for 24 hours the positive result was

considered as change of reagent color from red to yellow .
7. Motility test : a test tube contain semi solid nutrient agar was inoculated
with isolated bacteria by stabbing and incubate at 37 Co for 24 hours, the

spread of growth on the stabbing considered as positive result .
8. Urease test : the urea agar was streaked with isolated bacteria and
incubate at 37 Co for 24 hours, the change of media color from yellow to

rosy red considered as positive result (Cruickshank ,1975) .
3.2.1.5.5.Bacterial Identification by APi-20E

Another bacteriological test was performed to confirm the identification
of Gram negative bacteria according to instructions of company
3.2.1.5.6. Statistical Analysis
The results was analyzed statistically by chi square test and student's t
test to determine any significant variation between different means(Steel
and Torrie,1980).
Chapter four
Results
4. Results
4.1. Anatomical Study
4.1.1. Topography of Queen’s Reproductive System
4.1.1.1. Ovary
The results showed that the shape of the adult studied queen's ovary was
oval ,with slight bilateral compression ,its located in dorsal abdomen caudal
to the kidney ,figure4.1, and attached to the dorsal body wall by mesovarian
,to the diaphragm by suspensory ligament figure 4.2 and to the end of the
uterine horn by proper ligament of the ovary figure 4.3 .The ovary was
enclosed by bursa with a small slit- like opening located medially ,figure 4.4.
4.1.1.2. Uterine Tube
The uterine tube extends from the craniomedial aspect of the ovary to the
cranial ovarian end, and then extends caudally through the mesosalpinx to
join with the cranial end of the uterine horn, figure 4.5 and4.6.
4.1.1.3 Uterus
The uterus of queen was "Y” in shape with short uterine body and relatively
long uterine horns, figure 4.7.The uterine body was located beneath the
descending colon and above the urinary bladder, figure 4.8 .Each uterine
horn was extend cranially to meet the uterine tube ,the whole uterus (uterine
horn and uterine body) was attached to the body wall by the mesometrium.
The broad ligament ties the uterine horns to abdominal wall, formed by
folding the peritoneum, the round ligament of the uterus was imbedded in a
fold of broad ligament and extend caudolaterally to each near the inguinal
ring ,figure 4.9.
Kidney
Figure (4.1) reveal the position of ovary (arrow), caudal to kidney.
Figure (4.2) reveal the suspensory ligament (arrow)

Figure (4.3) reveal the proper ligament of the ovary(arrow)
Figure (4.4) reveal the ovarian bursa of the ovary(arrow)

CR
M L
R R
CD
Figure (4.5) reveal the topography of uterine tube, M:Medial;L:Lateral; CR:

Cranial ; CD : Caudal
Figure (4.6) reveal the unbend uterine tube

Figure (4.7) reveal the Y shape of the uterus
Bladder
Colon
Figure (4.8) reveal the uterine body (arrow)

B
B
.
Figure (4.9) reveal the broad ligament (B) and the round ligament (R)
4.1.2.Measurments of Queen’s Reproductive System
The measurements of pathological changes free reproductive system of 46

adult ,non pregnant queens were taken during the different seasons.
4.1.2.1.Ovary
During entire study period , the results showed that the mean of the
length, width and thickness of the right ovary were 9.8mm ±0.27 (mean
±SE; range 7-14.5) ,5.03mm ± 0.12(mean ±SE; range 3-8) and 3.95mm±
0.7 (mean ±SE; range 2-5) respectively ,while they were 9.56mm ± 0.26
(mean ±SE; range 7-14) ,4.8mm ±0.13(mean ±SE; range 3-7) and 3.05mm±
0.12 (mean ±SE; range 2-5) respectively in the left ovary . The statistical
analysis reveals no significant variation between right and left ovaries or
between the total mean and the means of the different seasons(P≥0.05)
,table4.1.
According to the age ,in the queens less than one year old , the mean of
the length ,width and thickness of the right ovary were 9.46mm ± 0.33(mean
±SE; range 7-12), 4.56± 0.18 (mean ±SE; range 3-5), 2.8± 0.12 (mean ±SE;
range 2-3.5) respectively, while they were 9 ± 0.33 (mean ±SE; range 7-
11.5), 4.64 ± 0.19 (mean ±SE; range 3-5), 2.48 ± 0.11 (mean ±SE; range 2-
3) in the left ovary . In queens above than one year old ,the length ,width and
thickness of the right ovary were 9.95 ± 0.38(mean ±SE; range 7-14.5) , 5.1
± 0.18 (mean ±SE; range 3.5-8) , 3.44 ± 0.14 (mean ±SE; range 2-5)
respectively, while in the left ovary they were 9.94 ± 0.35(mean ±SE; range
7-13.5) , 5 ± 0.19(mean ±SE; range 3.5-7) , 3.4 ± 0.13(mean ±SE; range 2-5)
respectively, table4.2.
4.1.2.2.Uterine Tube
The mean of the length of the right uterine tube was 54.4mm
±0.08(mean ± SE ; range 52.-60mm) while it was 55.7mm ±0.05 (mean ±
SE ; range 53-58mm) in the left uterine tube . The statistical analysis reveals
no significant variation between right and left uterine tubes and between the
total mean and the means of the different seasons ,table 4.3.
According to the age ,in the queens less than one year old , the mean of the
length of the right uterine tube was 53mm ± 0.33(mean ±SE; range 52-
54mm), while it was 56.8mm ± 0.33 (mean ±SE; range 53-61mm)in left
uterine tube. In the queens above than one year old ,the length of the right
uterine tube was 58.1mm ± 0.38(mean ±SE; range 51-65mm), while in the
left uterine tube it was54.4± 0.35(mean ±SE; range 50-60) ,table 4.4.
Table (4.1) reveal
the measurement
seasons
and right
ovaries during
M±SE
±0.70
±0.27
±0.12
±0.26
±0.13
±0.12
9.80
5.03
3.95
9.56
4.80
3.05
of leftTotal
different
46
46
46
46
46
46
n
Season winter Spring Summer Autumn
Ovary
n M±SE n M±SE n M±SE n Autumn
M±SE
Length 9.93 8.85 10.15 10.48

mm 15 ±0.46 10 ±0.17 10 ±0.21 11 ±0.63
±0.39
Width 5.13 5.20 4.80 4.46
mm 15 ±0.32 10 ±0.13 10 ±0.15 11 ±0.13
Right
Thickness 15 3.33 10 3.45 10 3.10 11 2.91
mm ±0.24 ±0.21 ±0.16 ±0.09
Length 9.53 8.90 9.60 10.85

mm 15 ±0.42 10 ±0.28 10 ±0.61 11 ±0.77
Width 4.90± 5.20 4.80 4.30

mm 15 0.29 10 ±0.19 10 ±0.15 11 ±0.16
Left
Thickness 15 3.00± 10 3.18 10 2.95 11 3.16
mm 0.26 ±0.25 ±0.15 ±0.10
Table (4.2) reveal the measurements(mm) of left and right ovaries according to
age
Age
Less than one Above than one year
ovary year
n M±SE n M±SE
Length 18 9.46±0.33 28 9.95±0.38

mm
Right Width 18 4.56±0.18 28 5.10±0.18
mm
Thickness 18 2.80±0.12 28 3.44±0.14
mm
Length 18 9.00±0.33 28 9.94±0.35
Left
Width 18 4.64 ±0.19 28 5.00±0.19
mm
Thickness 18 2.48±0.11 28 3.40±0.13
mm
Table (4.3) reveal the measurements(mm) of left and right uterine tubes during
different seasons
season Winter Spring Summer Autumn Total

n M±SE n M±SE n M±SE n M±SE n M±SE
U.tube
Right 15 56.50 10 55.60 10 57.70 11 55.30 46 54.40
±0.08 ±0.55 ±0.05 ±0.18 ±0.08
Left 15 55.90 10 55.90 10 55.10 11 56.30 46 55.70

± 0.08 ± 0.12 ±0.08 ±0.14 ±0.05
Table 4.4 reveal the measurements(mm) of left and right uterine tubes according
to age
Age
Less than one year Above than one year
U. tube n M±s n M±s

Right 18 53.00±0.33 28 58.10±0.38
Left 18 56.80±0.33 28 54.40±0.35
4.1.2.3.Uterine Horn
The results showed that the mean of the length and diameter of the right
uterine horn were 48.44mm ±1.47 (mean ± SE ; range 40-60) and 5.41mm
±0.5 (mean ± SE ; range 2-14) respectively ,while they were 54.43mm ±1.13
(mean ± SE ; range 42-65) and 51.4mm ± 0.08 (mean ± SE ; range 2-15)
respectively in the left uterine horn.
The statistical analysis reveals significant variation (P≤0.01) between the

measurements of right and left uterine horns ,and another significant
variation was observed between the total mean and each of the recorded
means of different seasons ,table 4.5.
M±SE
±1.47
±1.13
±0.08
48.44
54.43
±0.5
5.41
5.14
right uterine horns during
measurements of left and
Total
Table (4.5) reveal the
different seasons
46
46
46
46
n
M±SE
±3.34
±0.32
49.66
3.5±
0.34
3.48
3.58
54±
Autumn
11
11
11
11
n
M±SE
±2.12
±1.23
±2.15
±1.29
53.52
56.6
5.3
4.4
Summer
10
10
10
10
n
M±SE
±1.42
±1.04
±1.62
±1.16
44.8
7.45
7.7
50
Spring
10
10
10
10
n
M±SE
±1.73
±0.67
±1.86
±0.61
50.33
55.53
4.73
4.73
winter
15
15
15
15
n
Diameter
Diameter
Length
Length
mm
mm
mm
mm
U. horn
Season
Right
Left
The results also showed that in the animals less than one year old, the
mean of the length and diameter of the right uterine horn were 54.7mm±1.55
(mean ± SE ; range 42.5-62) ,3.38mm±4.83(mean ± SE ; range 2-14)
respectively, while they were 51.06mm±2.13 (mean ± SE ; range 42-65),4.5
mm ±0.95(mean ± SE ; range 3-15 ) respectively in the left uterine horn. In
animals above than one year old the mean of the length and diameter of the
right uterine horn were 53.57mm±1.38(mean ± SE ; range 42.5-62) ,6.4mm
±0.65(mean ± SE ; range 2-14) respectively, while they were 49.8mm ±1.44
(mean ± SE ; range 42-65),5.32 mm ±0.63(mean ± SE ; range 3-15 )
respectively in the left uterine horn. The statistical analysis reveals
significant variation (P≤0.01) between right and left uterine horns length,
while no significant variation observed between the means of uterine horns
lengths according to the age, table 4.6.
Table (4.6) reveals the measurement of length and diameter of left and right
uterine horns according to age
Age Less one year Above one year

U.horn
n M±SE n M±SE
Right Length 18 54.70±1.55 28 53.57±1.38

mm
Diameter 18 3.38±4.83 28 6.40±0.65
mm
Left Length 18 51.06±2.13 28 49.80±1.44
mm
Diameter 18 4.50±0.95 28 5.32±0.63
mm
4.1.2.4. Uterine Body
The results showed that the total mean of the length and diameter of the
uterine body were 22.75mm ± 1.52(mean ± SE; range 20-35) and 5.29± 0.36
(mean ± SE; range 3-12) respectively, Table 4.7 reveal the means of lengths
and diameters of the uterine body during different seasons. According to the
age ,the results showed that the length and diameter of the uterine body were
16.8mm ± 2.94(mean ± SE ; range 14-19) ,4.33 mm ± 0.57 respectively in
the animals less than one year and 26.53 mm ± 0.95 and 5.9 ± 0.58
respectively in the animals above than one year ,table, 4.8.
4.1.2.5. Cervix
The results showed that the mean of the length and diameter of the cervix
were 13.2mm ± 0.59(mean ± SE; range 6-20) and 8.72mm ± 1.95
respectively in the different seasons, table 4.9.
The mean ± standard error of the length and diameter of the cervix 12.6
mm ± 0.9 and 3.6 mm ± 0.44 respectively in the animals less than one year ,
and 12.26 mm ± 0.78 and 4.3 mm ± 0.35 respectively in the animals above
than one year, table 4.10.
Table (4.7) reveals the measurements of uterine body according to season
Season Winter Spring Summer Autumn Total
U.body n M±S n M±S n M±S n M±S n M±S
L 15 13.00 10 14.90 10 20.60 11 23.66 46 22.75

mm ±2.52 ±1.27 ±2.44 ±1.59 ±1.52
15 2.53 10 5.65 10 5.00± 11 2.41 46 5.29
D ±0.46 ±0.68 0.8 ±0.61 ±0.36
mm
Table (4.8) reveals the measurements of uterine body according to age

U.body
n M±SE n M±SE
L 16.80±2.94 26.53±0.95
mm 18 28
D 4.33±0.57 5.90±0.58
mm
Table (4.9) reveals the measurements of cervix according season
Season winter Spring Summer Autumn Total
cervix n M±S n M±S n M±S n M±S n M±S

Length 15 11.84 10 15.10 10 11.26 11 16.50 46 13.20
mm ±0.98 ±0.87 ±1.19 ±0.56 ±0.59
Diameter 15 5.50 10 6.80 10 4.25 11 6.51 46 8.72

mm ±0.31 ±0.64 ±0.59 ±0.18 ±1.95
Table (4.10) reveals the measurements (mm) of cervix according to age

cervix
n M±SE n M±SE
Length 18 12.60±0.90 28 12.26±0.78
mm
Diameter 18 3.60±0.44 28 4.03±0.35
mm
4.2. Physiological study

4.2.1. Follicular Phase
The number of animals with follicular phase was 14 out of 57of non
pregnant pathological changes free queens, in incidence rate 24.56%. The
highest incidence of follicular phase was in Winter 62.5%(5 / 8),followed by
Spring and Summer in percentage 44.4%(4 /9) and 33.3%(5 /15)
respectively, while no follicular activity was noted in animals during
Autumn, figure 4.10. According to age, all follicular activity was observed
in animals above 1 years old.
The size of recorded follicles was ranged from 0.5 to 2 mm, and the
number was ranged between 1- 3 in the same ovary, figure 4.11.
According to the months of study, the highest incidence of follicular phase

cases was in the February 100% and the lowest was in the August, while it
was absent in September, October and November and April ,figure 4.12.
70.00%
62.50%
60.00%
50.00%
44.44%
40.00%
30.00%
33.3
20.00%
10.00%
0%
0.00%
winter spring summer automn
Figure (4.10) reveal the incidence of follicular phase in different seasons
Figure (4.11) reveal the present of 2follicles on left ovary

120.00%
100%
100.00%
80.00%
60.00%
50% 40%
40.00% 40%
25%
20%
28.57% 16.66%
20.00%
0% 0% 0%
0.00% 0%
Figure (4.12) reveal the incidence of follicular phase in study months
Throughout the study period, out of 47 observed follicles,27 were present

in right ovaries and 20 in the left ovaries . Table 4.11 and figure 4.13 reveals
the incidence of follicles on the right and left ovaries.
Table (4.11) reveals the incidence of follicular phase according to individual ovary and
season
Season
Winter Spring Summer Autumn
Ovary
n 15 7 5 0
Right
% 60 53.85 55.55 0
n 10 6 4 0
Left
% 40 46.15 44.45 0
Total No. 25 13 9 0
70%
60%
60% 55.55%
53.85%
50% 46.15% 44.45%
40%
40%
Right ovary
30%
Left ovary
20%
10%
0% 0%
0%
Winter Spring Summer Autumn
Figure (4.13) reveal the incidence of follicular phase according to individual ovary
&season
The results showed that the uterus shape was differ according to size of
follicle\s ,the uteri of queens with follicles less than 1 mm were slightly
enlarged and congested, with straight uterine horns , figure 4.14 ,while the
uterine horns were twisted (Spiral in shape) with hyper folded if the follicles
larger than 1 mm ,figure 4.15.
The hormonal assay showed that the blood level of the estradiol -17ß in 14
follicular phase animals was 47.5pg/ml, ranged from 25-70pg/ml.
The percentage and range of vaginal epithelial cells in 14 estrus queens

for parabasal cells was0.92% (0-3) ,intermediate cells was 12.21 %(0-25),
nucleated superficial cells was 61.5 %(6-88) dna anucleated superficial cells
was 27.71%(4-88).
Figure (4. 14) reveals the straight uterus in follicular phase
Figure (4.15) reveal the spiral appearance of uterus in follicular phase

N
AN
Figure (4.16) reveal dominance of nucleated(N) and anucleated (AN) superficial cells in
vaginal smear of follicular phase
4.2.2. Luteal phase
The luteal phase was recognized in 17 out of 57 non pregnant

pathological change free queens, with incidence rate 29.82 %.
The highest incidence of luteal phase was in Spring 56.66%,followed by

Summer and Autumn in percentage 41% and 20% respectively, while it was
6.66% in Winter, figure 4.17. According to the months of study, the highest
incidence of luteal phase cases was in the April 80% and the lowest was in
the January 20%, while it was absent in February, October ,November and
December figure 4.18.
The average of corpora lutea per animal was 4.7 % ± 0.12 (Mean ± SE ;
range 2-5)with approximately equal distribution between right and left
ovaries as 2.41% ± 18(Mean ± SE ; range 0-5) in the right ovary vs. 2.35%±
0.58(Mean ± SE ; range 0-9) in the left ovary .The number of corpora lutea
per ovary was same in 3 queens (17,81%), greater in right ovary in 7 queens
(41.17%) and greater in left ovary in 7 queens (41.17%) .
60.00% 56.66%
50.00%
41%
40.00%
30.00%
20%
20.00%
10.00% 6.66%
0.00%
winter spring summer automn
Figure (4.17) reveal the incidence of luteal phase in different seasons

90.00%
80.00% 80%
70.00%
60.00% 50%
50.00% 50% 40%
40% 40%
40.00% 33.33%
30.00%
20.00% 20%
10.00% 0% 0%
0.00% 0%
0.00%
Figure (4.18) reveal the incidence of luteal phase in study months
The animals with this phase were gathered into 4 groups which describe as
following :
a) Recently ovulated queen:

In this study, only one queen was recorded as recently ovulated, it
has 2 corpus hemorrhagicum on the right ovary with 3 corpus
hemorrhagicum on the left ovary figure 4.19.
The oviduct and uterus were markedly enlarged and congested as
shown in figure 4.20.The estimated estradiol 81-ß and progesterone
were 16 pg/ml and 2ng/ml respectively.
b) Pregnant queens
The number of pregnant queens which noted in this study was 11/57 ,in
incidence rate 19.29%.The highest incidence of pregnancy was in Summer
33.33%,followed by Spring and Autumn in incidence rate 28.57% and 25%
respectively, while no pregnant cat was captured in Winter, figure 4.21.
According to months of our study, the highest incidence of pregnancy in

queen was in March 50%,followed by July ,September ,August ,June, April
and May in incidence rate 40% ,40%,33.33%,25%,20% and 20%
respectively, figure 4.22.
Figure (4.19) reveal the corpus hemorhhagicum in recently ovulated cat (arrow)
Figure (4.20) reveal the obvious enlargement of oviduct and uterus in recently
ovulated queen
35.00%
33%
30.00%
28.57%
25%
25.00%
20.00%
15.00%
10.00%
5.00%
0.00%
0.00%
Winter Spring Summer Automn
Figure (4.21) reveal the pregnancy percentage in different seasons

60.00%
50.00% 50%
40% 40%
40.00%
33.33%
30.00% 25%
20%
20%
20.00%
10.00%
0% 0% 0%
0% 0%
0.00%
Figure (4.22) reveal the pregnancy percentage in different months
The number of fetuses per each uterine horn was same in 7/11 queens
(63.63%),greater in right uterine horn in 2/11 queens (18.18%) and greater
in left uterine horn in 2/11 queens (18.18%).
According to 100% ovulation rate, the implantation rate was 86.06%

(range 50-100%).The implantation rate was 100% in 6/11 queens
(54.55%),80% in 1/11 queens ( 9.09%), 75% in 2/11 queens (18.18%),
66.66% in 1/11 queens (9.09%) and 50% in 1/11 queens (9.09%).Table 4.10
reveal the relationship between number of corpora lutea, number of fetuses
and implantation rate in 11 pregnant queens .
Table (4.10) reveals the relationship between numbers of corpora lutea, number of
fetuses
CL /animal No .of Average fetus Implantation

cats no.(range) rate %
2 3 2(-) 100
3 3 3(-) 100
4 2 3(2-4) 75
5 1 4(-) 80
6 1 5.33(4-6) 88.88
8 1 6.66(6-8) 83.33
The transuterine migration of embryos was observed in 3/11 queens with

incidence rate 27.27%. All transuterine migration of embryos was occurred
in animals with 100% implantation rate.
The maximum number of embryos that migrate per animal was 2 ,in the
first queen , two embryos were migrated from right to left uterine horn , in
the second queen one embryo migrated from right to left uterine horns
,while in the third queen one embryo was migrated from left to right. figure
23.4.
In all animals with transuterine migration of embryos the difference in

number of fetuses between left and right uterine horns to the difference of
corpora lutea between left and right ovaries were 1:3 ,0:2 and 2: 4 .
2embryo
3CL
Figure (4.23) reveal the migration of 2 embryos from right to left uterine horn
C ) Pseudopregnant queens
The pseudopregnancy was noted in (2/17) queens (11.76 %) , the first
queen has only 2 active corpora lutea on the right ovary , while the other
queen has 2 active corpora lutea on each ovary , the uterus and the uterine
tube enlarged slightly. The means of length and diameter of right and left
uterine horn uterine were 45 × 8 mm and 47.5 ×8 mm respectively, while
the means of length and diameter of uterine body were 21.25 × 6.5 mm
respectively, figure 4.24.
In pregnant and pseudopregnant animals, the mean of plasma

progesterone concentration was 48 ng/ml (range 22-56) ,and the cell
comprising the vaginal smear in diestrus cats (mean percentage followed by
range) included intermediate cells ( 47% ; 18-78) parabasal cells(45%;12-
82), anucleated superficial cells (0%),nucleated cells (5%; 0-5) ,figure 4.25.
D)Queens with involuted uterus
The involution was noted in 3 /17 with incidence rate 17.64% ,all queens
were lactating , and had inactive corpora lutea on their ovaries ,the uteri
were straight and markedly dilated ,figure 26.4 and 27.4.
The mean ±SE of length and diameter of right and left uterine horn were
45.66× 12mm and 40×11.66 mm respectively , while the means of length
and diameter of uterine body were 21× 6.66 mm respectively.
Figure 24.4 reveal the appearance of reproductive system in pseudopregnancy
Figure( 4.25) reveal vaginal cytology during the diestrus phase

Figure( 4.26) reveal the appearance of involuted uterus
Figure (4.27) reveal the inactive C.L. of involuted reproductive system(A),compare

with active C.L. (B)
4.2.3. Anestrus phase

The anestrus phase was recognized in 26 out of 57 pathological free
queens , with incidence rate 36.84% .The highest incidence of anestrus
phase was noted in Autumn 86.66%, followed by Winter ,Summer and
Spring in incidence rate 46.66% ,20% and 7.14% respectively, figure 28.4.
According to the months , the highest incidence of the anestrus phase cases
was in October and November 100% and the lowest was in May and July
20% ,while it was absent in February and March ,figure 29.4.
100.00%
90.00% 86.66%
80.00%
70.00%
60.00%
50.00% 46.66%
40.00%
30.00%
20.00% 20%
10.00%
7.14%
0.00%
Winter Spring Summer Automn
Figure (4. 28) reveal the mean of the anestrus phase according to the seasons
120.00%
100% 100%
100.00%
80.00% 0%
60%
60.00% 50.00%
40% 40%
40.00%
25%
20% 20%
20.00%
0%
0.00% 0%
Figure (4.29) reveal the anestus phase according to the months
In all observed animals with anestrus phase the ovaries were entirely
smooth ,and the mean of uterine horns measurements was at the minimum
of all recorded measurements figure 30.4 and 31.4.The mean ±SE of
length and diameter of right and left uterine horn were 50.5 ±1.54 × 3.2±
0.6mm and 54.65±1.87 ×11.66 ±1.45mm respectively ,while the means±SE
of length and diameter of uterine body were 16.2±2.73×4.5±0.45
respectively .
Cells comprising the vaginal smear in anestrus queens (Mean percentage

followed by range )included intermediate cells 87.9%(50-100%), parabasal
cells 8.6% (0-50%) ,nucleated superficial cells 2.7%(0-22%) , anucleated
superficial cells 0.8%(0-2%) ,figure 32.4.
.
Figure( 4.30) reveal the reproductive system during the anestrus
Figure (4.31) reveal the reproductive system during the anestrus

Figure( 4.32) reveal vaginal cytology during anestrus phase
4.3.Pathological Study
During the study period, out of 06 only 3 queens with gross reproductive
systems abnormalities were observed ,with incidence rate 5% ,these cases
were described as following :
A. Case (1):Parovarian(Periovarian) Cyst
One queen with parovarian cyst was captured in this study, this queen was
above one year old and weigh 5 kg ,the clinical examination revealed that
she was healthy. After dissecting the animal a large parovarian cyst was
located on the left ovary , it was lobulated, thin walled cyst filled with clear
fluid (about 6ml) ,this cyst was surround the ovary entirely, figure 4.33 and
4.34. The ovary and uterine horns was normal grossly .The right ovary was
with previous ovarian activity ,with 1 small follicle on its surface.
Parovarian cyst
oviduct
Figure (4.33) reveal lobulated parovarian cyst
Figure (4.34) reveal parovarian cyst(arrow)

Table(4.13)reveal the measurements of reproductive system with parovarian cyst
Part Measurement/mm
Ovary Right 7 ×5×3
(L×W×Th)mm Left 7×4×3
Oviduct(L )mmRight 50
Left 55
U.horn Right 60×6
(L×D)mm Left 75×9
U.body(L×D)mm - 25×5
Cervix (L×D)mm - 15×9
The histpathological examination of the ovary with the parovarian cyst

reveal normal microscopical structure of the ovary and oviduct .
B. Case (2): Follicular Cyst
Only one queen in this study was reported with follicular cyst. The queen
was over one year old and weight about 4.5kg
The clinical examination revealed that the queen was healthy , after
dissecting the animal ,a follicular cyst was found on the left ovary ,thin
walled cyst ,filled with clear fluid ,5mm in diameter , the ovaries and
uterine horns were normal grossly ,Figure 4.35.
Figure (4.35) reveal the follicular cysts(arrow)
Tables (4.14) reveal the measurement of the reproductive system with follicular
cyst
Part Measurement/ml
Ovary Right 9×5×4
(L×W×Th)mm Left 7×5×4
Oviduct(L )mm Right 4.5
Left 4.8
U.horn Right 40×5
(L×D)mm Left 45×5
U.body(L×D)mm 24×7
Cervix (L×D)mm 17×8
The histopathological examination of the ovary with the follicular cyst

reveal normal micrscopical structure of the ovary with presence fibrosis and
relatively large follicles as well as normal sized follicles .
C. Case (3): Pyometra
The third case which reported in this study was pyometra , the captured
queen was above one year old and weight about 4kg , the clinical
examination revealed that the queen was healthy , after dissecting the animal
,the uterus was enlarged with congested appearance ,figure 4.37 and 4.38.
The color of the two uterine horns was abnormal and the endometrium was
very congested with present of exudates.
Congested uterus
.
Figure (4.37) reveal the abnormal appearance of uterus with pyometra grossly
Figure (4.38) reveal the marked thickness of the endometrium and presence of cheesy
pieces(arrow)
The histopathological examination of the uterus with the endometritis

reveal sever infiltration of neutrophils in sub epithelium and between the
uterine glands ,large number of necrotic neutrophils in the lumen of the
uterine glands and uterine lumen and the exfoliation of the epithelium, figure
4.39.
The histopathological appearance of the uterine tube reveal the dilation of

the uterine tube and epithelial atrophy , infiltration of the epithelium , fusion
of villi formed vacuole filled with pus and mucous ,the lumen of the uterine
tube filled with mucous 4.40.
The bacterial isolation and identification of exudates culturing reveals 2

isolates ,one is G-ve bacteria ,the growing on selective media, features of
colony and biochemical test reveal the characteristics of E.coli ,the another
G+ve isolates is confirmed as Staphylococcus spp.
Figure (4.39) reveal sever infiltration of neutrophils in sub epithelium and between the
uterine glands ,large number of necrotic neutrophils in the lumen of the uterine glands and
uterine lumen and the exfoliation of the epithelium (arrow),(H&E.X200)
B A
Figure (4.40) (A&B) reveal the dilation of the uterine tube fusion of villi formed vacuole
filled with pus and mucous(H&E.A.X200,BX100).
Chapter five
Discussion
5. Discussion
5.1. Anatomical Study
5.1.1. Topography of Genital Parts:
All the obtained topographical features, include the location and

attachment of different genital parts of non pregnant pathological changes
free Iraqi feral queens are completely compatible with other results were
recorded by many authors (Del Campo and Ginther ,1974 ; Shille et
al.,1983;Johnston et al.,2001 ; Chatdarong et al.,2005).
5.1.2. Measurements
Out of 40 of non pregnant gross pathological changes free queens the

mean of measurements (length × width ×thickness ) of right and left ovaries
were 9.8mm ± 0.27 × 5.03mm ± 0.12× 3.95mm ± 0.7 and 9.56mm ± 0.26
× 4.8mm ±0.13 ×3.05mm± 0.12 respectively. There were no significant
variations between the total mean and the mean of each Winter, Spring,
Summer and Autumn, as shown in table 4.1.The measurements of queen's
ovaries in this study are close to that obtained by Johnston et al., (2001)
(10mm length ×3mm width 5mm thickness).
The absence of significant variation between the measurements of age

classes are in disagreement with Bliley &Al-Qara’awy ,(2005), that they
supposed that the length and width of ovaries of young queens were
3mm×2mm respectively , while in adult queens were 10mm×5mm. The
reason of the disagreement may be due to that the authors may include
immature queens, while we study the measurements in mature queens aged
less than 1 year, or due to breed variations none of other references refer to
any variation between right and left mature queen's ovaries on age basis.
The means of length of right and left uterine tubes in this study were
54.4mm ±0.08 and 55.7mm ±0.05 respectively with no significant variation
between the right and left uterine tube according to season or age. Similar
results were obtained by Fletcher, (1974), who’s reported the range of
uterine tube length (50-60mm).
Also, our results showed that the means of length and diameter of right
uterine horn were 48.44mm ±1.47 and 5.41mm ±0.5 respectively, while they
were 54.43mm ±1.13 and 5.14mm ± 0.08, respectively in the left uterine
horn, with significant variation between the measurements of right and left
uterine horns, and between the total mean and each of the recorded mean of
different seasons.
Non of any reference refer to difference in the length of right and left
uterine horn, but Al-Delemi, (2007), recorded same results in Arabian she
camels which has the same 'Y' shape uterus, Bliley &Al-Qara’awy, (2005)
explained the same findings as that incidence of pregnancy in left uterine
horn is greater than right one, and the confirmation of this result as fact in
cats, need more studies of more samples either live or postmortem
examination.
The results showed that the total means of the length and diameter of the
uterine body were 22.75mm ± 1.52and 5.29± 0.36 respectively with
significant variation between the total mean and the means of different
season .According to the age, the length and diameter of the uterine body
were 16.8mm ± 2.94 and 4.33 mm ± 0.57 respectively in the animals less
than one year and 26.53 mm ± 0.95 and 5.9 ± 0.58 respectively in the
animals above than one year. The length and diameter of the cervix were
13.2mm ± 0.59 and 8.72mm ± 1.95 respectively with significant variation
between the total mean and the means of different season. the mean ±
standard error of the length and diameter of the cervix 12.6 mm ± 0.9 , 3.6
mm ± 0.44 respectively in the animals less than one year , and 12.26 mm ±
0.78 , 4.03 mm ± 0.35 respectively in the animals above than one year . The
seasonal variation in our data are in agreement with result of Reece, (1997),
as fact that the uterine horn is place of implantation, so it is influenced with
the physiological sequences of estrous phase, and that interpret that the
lowest measurements in this study was recorded in season of anestrous
(Autumn and Winter), while the variation in measurement was observed in
other seasons according to the incidence of different reproductive stages.
The measurements of uterine body and cervix are similar to those reported
by Bliley &Al-Qara’awy, (2005) and Johnston et al., (2001)
5.2. Physiological Study
5. 2.1. Incidence of Sexual Activity (Follicular and Luteal Phases)
Although several researchers refer to that in the tropical areas ,the

domestic cat may breed throughout the year(Little,2001 ;Simpson et al.,2004
;Fontbonne and Malandain,2006 ),there are several factors may influence or
determine breeding of domestic cat include length of day, environmental
temperature and breed(Eldredge et al.,2008),so the fluctuation in breeding
may vary from country to country .Fleldman and Nelson,(1996) reported
that in temperate zone the high environmental temperature may decrease the
estrous incidence ,while Shille and Sojka,(1995) revealed that the effect of
heat and humidity on domestic cat breeding depend on individual variation.
The present study was preformed along different 4 seasons on feral cat,
which they are roaming outdoor and expose to natural irregular light as well
as possibility of high individual variations existence, the fluctuation in
incidence of follicular and luteal phases was frankly observed. Therefore in
this study the results showed that 14 out of 57 pathological free queens were
observed with follicular phase, in percentage rate 24.65%.Figure 4.10
demonstrate the distribution of follicular phase incidence in different
seasons. The highest incidence was recorded in Winter(62.5%) ,followed by
Spring(44.44%) and Summer(20%) ,while no follicular activity was
observed in Autumn.
on month basis, Figure 4.11 reveal that the highest incidence of follicular
phase was recorded in February (100%), While it was absent in April,
September ,October and. And November .
Also in this study, the incidence of luteal phase was 29.82 %, and the
highest incidence was in Spring(56.66%) and the lowest was in
winter(6.66%),and according to months, figure 18.4 reveal that the highest
incidence of luteal phase was in April(80%) and the lowest was in January
(20%),while it absent in December, February ,October and November .
5.2.2 Incidence of Sexual Inactivity (Anestrous Phase)
The incidence rate of anestrous phase which observed in this study was
45.6% ( 26 out of 57), figure 4.28 reveal that the highest incidence of
anestrous phase was noted in Autumn 86.66%, followed by Winter ,Summer
and Spring in incidence rate 46.66% ,20% and 7.14% respectively, and
according to the months , the highest incidence of the anestrous phase was in
October 100% and the lowest was in May and July 20% ,while it was absent
in February and March .
The highest incidence period of anestrous in this study are in agreement

with Johnston et al., (2001), that said it occurs in late Autumn months
include October, November and December .Simpson et al., (2004) explained
this as result of day light shortage under natural conditions.
The ovarian inactivity in Winter may be due to elevation of Prolactin and

melatonin in circulating plasma(; Leyva et al.,1984 ;Leyva et
al.,1989;Simpson et al.,2004 ),while the relatively high incidence of
anestrous phase in Summer may be due to increase of the environmental
temperature as recorded by (Concannon and Lein ,1983).
5.2.3. Pregnancy and Pseudopregnancy
The number of pregnant queens which noted in this study was 11/57 ,in
incidence rate 19.29% and the highest incidence of pregnancy was recorded
in Summer 33.33%,followed by Spring and Autumn in incidence rate
28.57% and 25% respectively, while no pregnant cat was captured in Winter
and according to months of our study, the highest incidence of pregnancy in
queen was in March 50%,followed by July ,September ,August ,June, April
and May in incidence rate 40% ,40%,33.33%,25%,20% and 20%
respectively, figure 4.22 , these data are in agreement with pope, (2000)
who reported that under the natural conditions the free ranging domestic cats
can produce occasionally 3 litters /year, with 1-5 kitten in each litter, and the
incidence of pregnancy is in marked contrast to pregnancy of bitch, which
may breed only twice a year (Feldman and Nelson ,2004; Simpson et
al.,2004)
In this study ,the number of fetuses per each uterine horn, implantation
rate, relationship between number of corpora lutea, number of fetuses,
transuterine migration of embryos and the maximum number of embryos
that migrate per animal are in agreement with Tsutsui et al., (1989) they
observed that the fetal number was tend to distributed equally between the
left and right uterine horn and implantation rate increased when the ovaries
contain 1-8 C.L. The transuterine migration of embryos was observed in
3/11 queens with incidence rate 27.27%; all transuterine migration of
embryos was occurred in animals with 100% implantation rate.
Little references are available about transuterine migration of embryos in

domestic cat ,but it has been recorded in other litter bearing animals ,Dziuk
et al.,(1964) reported it in 40-50% of pregnant sows, while Tsutsui,(1975)
observed it in 50% of pregnant bitches. All previous our data are near to that
of Tsutsui et al., (1989) who recorded that the transutrine migration of
embryos occurred from the side with ovary containing the greater number of
CL to the side of fewer CL and the migration of embryos more than 3
embryo was not observed, while the migration rate recorded by the above
authors was more than that recorded in this study, may be due to the
difference in the number of examined cats between studies.
Feldman&Nelson,(2004) reported that the queens may undergo as many as

4-5 pseudopregnancies during the course of one poly estrus season . Michael
et al., (2008) observed enlargement of abdomen with increase of appetite
and weight gains in queens with pseudopregnancy, while Concannon& Lein,
(1983) revealed that the average of duration of luteal activity in
pseudopregnancy was 36.5 days. All these data are close to that obtained in
present study where the pseudopregnancy was noted in 2out of 17(11.76 %)
queens with at least 2 active corpora lutea with slightly enlargement of
uterus and the uterine tube.
5.2.6 Vaginal Cytology
The cyclical changes of vaginal exfoliated epithelium in canine have been

extensively studied and are widely used clinically (Mills et al., 1979;
Simpson et al., 2004), while only few reports are available on vaginal
cytology in feline species.
In the present study, the morphological changes of vaginal epithelium

during the various stages of estrous cycle were reported in the estrus phase,
out of 14 estrus queens, the percentage of vaginal epithelial cells reveal that
the dominant cells were superficial including nucleated superficial cells
(61.5 %) dna anucleated superficial cells( 27.71% ).The parabasal cells and
intermediate cells were 0.92% and 0.25% respectively. These data are
similar with that obtained by Johnston et al., (2001) and Shille et al., (1979)
documented that the number of superficial cells is fairly constant throughout
behavior estrous and ranged from 40-60% of total cells present.
In the diestrus queens ,the cell comprising the vaginal smear were include
intermediate cells ( 47%), parabasal cells(45%) and (5%) and this findings
are in agreement with that of Tsutsui &Stabenfeldt,(1993) reported 50%
,48% and 2% for intermediate cells , parabasal cells and superficial
nucleated cells respectively.
Also, the rate of cells comprising the vaginal smear in anestrous queens
were include intermediate cells 87.9% , parabasal cells 8.6%,nucleated
superficial cells 2.7% and anucleated superficial cells 0.8%(0-2%) are
incompatible with finding of Mills et al.,(1979) noted that during anoestrus
the cells are rare , mucus is obvious and a bund ,and the majority of cells are
basal or parabasal with some intermediate cells, while the present results are
in agreement with Johnston et al.,(2001) described that the exfoliated
vaginal epithelial cells consist of less than 10% parabasal cells,40%to70%
intermediate cells , and 30% to 40% nucleated superficial cells ,background
debris is obvious .
5.2.7. Hormonal Changes
The hormonal assay in present study was revealed that the highest mean
of serum estradiol was recorded in the follicular phase 47.5 pg/ml, ranged
from 25- 70 pg/ml and the level was 16 pg/ml during the recently ovulated
queen, these finding are in agreement with Long,(2006) who recorded a
concentration of 20-50 or higher during proestrous and estrus phases, and
within the range of Feldman and Nelson,(2004) who observed that the level
of serum estradiol was increase from 25 pg/ml in first day of estrus to the
peak 50 pg/ml in 5th day, then decline to 10 pg/ml in the10th day.
During the luteal phase the mean of progesterone level was 2 ng/ml in
recently ovulated cat (with obvious corpus hemoragicum), increase to reach
48 ng/ml (range 22-56) in animals with diestrus (pregnancy and
pseudopregnancy),these results are in disagreement with Johnston et al.,
(2001) they suspected the serum progesterone concentrations ranged from
1.5ng/ml to more than 20 ng/ml in the queens with diestrus.
All examined blood samples which obtained from the queens in anestrous
phase reveal that the level of estrogen and progesterone at the basal level,
these are compatible with levels mentioned by others (Johnston et al., 2001;
Simpson et al., 2004).
5.3. Pathological Study
The relatively low incidence of reproductive problems in present study

(5%) might be due to small sample size, or due to fact, that the feral cat has
short life comparable with household queens which receive more care and
hygienic conditions.
In present study, only one case was reported of single, lobulated and thin
walled cyst filled with clear fluid parovarian cyst, the ovaries and uterine
horns were normal grossly and microscopically.
There were no available reference refer to recording of parovarian cyst in

cat ,except McEntee, (1990) explained that it arises either from remnant of
the mesonephric (walffian) tubules or the paramesonephric (Mullarian)
tubules. Noaks, et al.,(2003) suggested that many tiny parovarian cysts
ranged 1-3cm in diameter, are very common incidental finding in
slaughtered cattle. Al-Delemi, (2007) recorded two cases of parovarian cysts
in Iraqi she camel.
The size and description of follicular cyst was found in present study
was in agreement with Feldman and Nelson ,(2004) and McEntee,(1990)
they thought that the normal follicles of the queens is up 2-3mm in diameter
and the larger follicles are said to be cystic, and with Gelberg et al.,(1984)
whose observed cysts as unilateral in 16 cats and bilateral in four from 20
cats .
Potter et al.,(1991) concluded that the prevalence of endometritis in cats
increases with age in sexually intact female cats and mainly after parturition
,these close to case was reported in this study , which was lactating ,over 1
years old queen .All observed gross and microscopical changes were
compatible with description of Potter et al.,(1991) and Misirlioglu et
al.,(2006) .
The isolates of concurrent case (E.coli and Staphylococcus spp.) were in

agreement with results of Davidson et al., (1992) and Kenney et al.,(1987).
Patricia et al.,(1999) reported E.coli and Staphylococcus spp., as vaginal
flora in various stages of queen estrous.
Chapter six
Conclusion & Recommendation

Conclusions
It is concluded that:
1. The present study considers only one of its kind in Iraq which
includes some anatomical , physiological and pathological aspects in
Iraqi feral queens reproductive systems.
2. The Iraqi feral queens are poly estrous, breeding throughout the year.
3. Only the uterine horn ,uterine body and cervix measurements are
influenced by season
4. The highest incidence of follicular phase in Winter and the lowest in
Summer , while it was absent in Autumn , with increase of the
estradiol 17-ß to 25-70pg /ml and dominance of superficial cells in
vaginal cytology .
5. The incidence of the luteal phase is high in Spring and
characterized by high serum level of progesterone with dominance of
intermediate cells in vaginal smear.
6. The anestrus phase is recorded in all seasons, with minimal
measurements of genital parts .
7. The highest incidence of pregnancy is in Summer and the number of
recorded fetuses/ animal ranged 2-8 .
8. The implantation rate is 86.06% ,and transuterine migration of
embryos occur in 27.27% , the maximum number of embryos that
migrate per animal was 2.
9. The study demonstrate low incidence (5%) of pathological changes in
feral queens reproductive systems ,the recorded cases are include
parovarian cyst ,follicular cyst and suppuative endometritis .
10.The vaginal smears assist in determination of feline various estrous
phases .
11.The estimation of serum progesterone can not differentiate between
pregnancy and pseudopregnancy ,while relaxin consider as indicator
for pregnancy .
Recommendations
It is recommended that:
1. control of the onset of feline puberty by More studies which
carried out about the environmental and physiological conditions.
2. Applying of advanced procedures in feline reproduction such as
ultrasonic device and fluoroscopy .
3. More researches are to be done about the infertility in domestic
cats .
4. Carrying out the same present study on Iraqi middle sized wild
cats ,to determine the steps of wildcat preservation.
5. More researches are to be done about the reproductive systems of
tom cat of feral cats.
Chapter seven
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reproductive physiology in queen (cat)

Presentation · December 2017

Asst. Prof. Hala Jawad Alfatlawy

infertility in mare causes and treatment View project

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The user has requested enhancement of the downloaded file.

Asst .Prof. Dhia H.Jassim,D.Sc Asst. Prof. Thaier A.Abid,

Asst .Prof. Jabbar A.A. AL-Saaidi , D.Sc

Dr. Mohammad J. Alwan

College of Vet. Med., University of Baghdad

Dr.Ihsan Hammody Saeed Dr. Taher Abdl- Hussain

Asst. Prof.(member) Asst. Prof.(member)

College of Vet. Med., College of Vet. Med.,

University of Baghdad University of Basra

Asst. Prof.(member and supervisor) Asst. Prof.(member and

College of Vet. Med., College of Vet. Med.,

AL-Qadisiya university AL-Qadisiya university

Approved by the council of the college of the Veterinary Medicine

Asst.Prof. Dhia H. Jassim Prof. Dr. Khairi

Head of department the Dean

Date of Examination : 27/6/2010

To ..My Beloved Husband ,Dr. Haider Al-

2.2.1.4.2. Blood Supply 9

2.2.2.6.4. Hormonal Changes 28

3.1.2.3. Female Reproductive Tract Samples 38

4.21 The Pregnancye in Different Seasons 71

In past, the cats are used as familiar symbols of independence, power,

1. Study of topography and measurements of queen's non pregnant

2.1. The Cats

2.1.1.Scientific Classification of The Domestic Cats

The Scientific classification of domestic cat mentioned in table 2.1

Sub kingdom Eumetazoa

2.1.3. Hybridation with Wild Cats

Table (2.2) reveals the Domestic cat's breeds

Breed Country Origin Coat

Abyssinian Egypt Natural short

American Curl United States Mutation Short/long

American Wirehair United States Mutation Rex

Burmese Burma Natural Short

California Spangled California Crossbreed Short

Devon Rex England Mutation Rex

Egyptian Mau Egypt Natural Short

Javanese - Crossbreed Long

Manx Isle of Man Natural/mutation Short/long

Persian Iran - Long

Peterbald Russia Cross breed Hairless

Russian Blue Russia Natural Short

Siamese Thailand Natural Short

Somali Somalia Mutation Long

Turkish Angora Turkey Natural Semi long

2.2.1. Anatomy of Female Genital Tract

2.2.1.2.1. Morphology and Location

The ovary of adult queen is oval in shape with slightly compressed

As contrast to most large animal ,the feline ovaries are located in

2.2.1.2.2. Blood Supply

2.2.1.2.3. Ovarian Follicles

The ovarian follicles is a small simple structure appear in spherical shape

2.2.1.2.4. Corpous Luteum

2.2.1.3 Uterine Tubes (oviducts)