Ar 405 Clinic Final

REPRODUCTION CLINIC - II
(AR 405)
To the Point Class Lectures

Delivered in Practical Classes
FINAL EXAM NOTES COMPLETE*

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Presented by:
MUHAMMAD SAJJAD HUSSAIN
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Up to Date: 25 May 2011 W ednesday
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* Only Portion of Dr.Nafees Akhtar is missing in this handout. Please consult the given handouts for all topics covered by said teacher. These handouts are available at photostat shop on your request. Thanks !!!
Delivered by: Dr. Najib Ur Rehman Clinical Use of Reproductive Hormones

Hormones are organic physiological substances. All reproductive processes are driven by hormones. Hormone: Hormone is a chemical substance that regulates certain activity of its target tissue (target is specific tissue which has receptors for the effect of the hormone). Receptor: Receptor is a special molecule present on the cell that binds with hormone to bring the required effect of the hormone. Hormones are organic physiological substances. All reproductive processes are driven by hormones. When problem is there in the effect of hormone function then it also indicates that the problem may lie due to the number of receptors present at target site.
GnRH (Gonadotropin Releasing Hormone)

GnRH is the mother reproductive hormone. It is a deca-peptide hormone. It is secreted from hypothalamus which is 1/300 part of the brain. It is the part of diencephalon present at the base of the brain. Hypothalamus has three areas: a) Preoptic nucleus, b) Suprachiasmatic nucleus (SCN), c) Anterior hypothalamic area (AHA). Hypothalamus is made up of neurons, secretion occur from cluster of cell bodies and gonadotrops in the pituitary are the target cells. It controls hunger, anger, emotions and sexual process and pleasure.
Factors which STIMULATE GnRH secretions

These factors can be divided into two categories, viz.: External or environmental and Internal or endogenous. External Factors: Good balanced nutrition Viewing sexually attractive individual of opposite sex Weather (pleasant) especially in seasonal breeders. Long day is favourable for mare. Short day is favourable for sheep. Favourable, comfortable and stress free environment. Internal Factors: Epinephrine & Nor-epinephrine. Melatonin favours the release of GnRH Coitus in Cat (physical stimulation to vagina) In cats and camels the sexual stimulation favors the release of GnRH as they are induced ovulators. Progesterone (P4) and Estrogen (E2) in acyclic females. Sight and Hearing of sexually attractive individuals. (especially for males) Pheromones favor the release of GnRH.
Factors which SUPPRESS GnRH secretions:

External Factors: Stress and Depression Starvation Prolonged sickness and Acute pain Internal Factors: Prolactin (a hormone) suppress the secretion of GnRH. Opoids i.e. -endorphins, secreted while suckling, thus suppress the release of GnRH Progesterone (P4) will lead to low GnRH production. Low Estrogen (estradiol 17) will lead to low production of GnRH. Estradiol 17 is needed for animal to be in heat.
Clinical Uses of GnRH

Managemental Uses of GnRH To enhance puberty in pre-pubertal age heifers. (To bring puberty earlier). To induce cyclicity in post-pubertal age heifers, if ovaries are of normal size. To induce cyclicity in postpartum females or early ovarian rebounce in postpartum females. To minimize the incidence of follicular cysts in postpartum females. In high yielder cows, incidence of follicular cysts is high in postpartum period; due to low LH secretion. So all postpartum females must receive single shot pf GnRH 20 days after parturition. Follicle on ovary ---> estrogen ---> LH will release upon GnRH stimulation. No follicle on ovary ---- > estrogen --- > FSH will release upon GnRH stimulation To improve conception rate, give a single shot of GnRH at the time of AI. It will cause release of LH surge and ovulation will occur at time, so improve the conception rate. There is about 5-10 % increase in conception rate. To improve pregnancy rate, give a single shot of GnRH 10-12 days post-AI. It will cause release of LH which ultimately acts on luteal cells of CL in order to stimulate the activity of CL for progesterone release. It is used in heat synchronization regime. Clinical Uses of GnRH To treat ovarian follicular cyst To curtail the prolonged heat period in mare. GnRH will cause release of LH, thus ovulation will occur. For this purpose, it is injected at 5th day of heat (estrus). To curtail the prolonged proestrus in bitch. In proestrus, bitch discharged blood out of vulva, so heat comes when blood discharge from vulva stops and female start accepting the male. Proestrus period in bitch is normally 10-12 days. In bitch, certain level of progesterone is also needed to come in heat, so inject GnRH after 10 days of proestrus, it will cause release of LH which ultimately luteinize the follicular cells and start releasing progesterone (Normally luteinization begins before ovulation in bitch).
To treat the cases of delayed ovulation; because of property of GnRH to release bulk amount of LH hormone. Fertlization failure is because of delayed ovulation, so a single shot of GnRH at the time of AI or service will solve this problem and increase the fertilization rate.
Products of GnRH
These are categorized into two types: i) Natural GnRH ii) Synthetic GnRH
Natural GnRH Preparations

It is very difficult to get natural GnRH because of its so short half life. No clinical use has been reported, synthetic preparations are only available.
Synthetic GnRH Preparations

>> Gonadrolin: Its structure is similar to that of natural GnRH. It is available with different trade names in the market: Cystorelin and Factel [50 g/ml] >> GnRH analogues: Such preparations are: Buserilin [Receptal 4 g/ml by Hoest; Conceptal 4 g/ml by Star], Lecirelin [Dalmarelin 25 g/ml] Dose rate: >> Gonadrolin: Normal dose(for post AI or service, and follicular growth etc.): 250 g/cow IM Dose for treatment of follicular cyst: 500 g/ml IM >> Buserilin: Normal dose (for post AI or service and follicular growth etc.): 20 g/cow IM Dose for treatment of follicular cyst: 40 g/cow IM >> Lecirelin: Normal dose( for post AI or service and follicular growth etc.): 50 g/cow IM Dose for treatment of follicular cyst: 100 g/cow IM Precaution: You must wait for 8-10 days after treatment. If no effect, then repeat it. You also do attention on nutrition of animal from 1 month before treatment. Protein and mineral mixtures must be provided to animal in their ration. Animal with poor body condition usually does not respond optimally to hormonal treatment.
Gonadotropins
These are of two types: i) Pituitary gonadotropins: FSH & LH ii) Placental gonadotropins: hCG & eCG FSH, LH, eCG and hCG are gonadotropic hormones. These are members of family glycoprotein.These hormones are made up of alpha and beta carbohydrates molecule chains. These two chains are linked by covalent bonds. Alpha subunit is same in all these four hormones having 92 amino acids but beta subunit is different which is responsible for
biological function. But alpha is also needed. Without a subunit the structure is incomplete and can not perform its function. Half life of Gonadotropins: Pituitary gonadotropins: FSH: 2 hours Placental gonadotropins: eCG: >3 days LH: 2 hours hCG: >12 hours
Comparative Features of pituitary and placental gonadotropins

Pituitary Gonadotropins (FSH, LH) Less half life Costly Low CH2O contents Placental Gonadotropins (eCG, hCG) Long half life Cheap High CH2O contents
FSH: Follicotopin Growth of follicle from secondary to tertiary follicle It plays major role in synthesis of estradiole Target is ovary Source of natural FSH is pituitary of porcine or bovine. It is also produced now a days through genetic engineering by inserting FSH producing gene into the E.coli organism. LH: Final growth of follicle, ovulation, formation of CL, maintenance of CL. Target is ovary and CL. Source of natural LH is porcine or bovine pituitary. eCG/PMSG: Its action is like FSH in all animals except in mare where its action is LH like. It is released in mare at 40 day pregnancy by endometrial cups (specific structure formed by trophoblast cells of embryo) by day 70 of pregnancy PMSG reaches to peak level i.e. 100 IU per ml of plasma and this peak persist up to 3rd month. During 4th month capacity of production reduces gradually and in 5th month it stops. No PMSG as endometrial cups are destroyed (destruction start in 4th but completed in 5th month). Fetus is 50% foreign for mother. In uterus local immune system becomes weak that it does not attack fetus. Pg weakens local immune system. Maternal system recognizes endometrial cups, kill them gradually, upto 5th month all dead no PMSG. hCG: Acton is LH like. Embryo starts production of HCG by day 10th of pregnancy because it implements with endometrium by that time. The outermost cells (trophoblast) start producing hCG. At day 90 the peak production of hCG is seen. At 5th month its production decreases but low level 22 IU per ml of plasma of hCG continues to be produced in it till end of pregnancy. It is leuteolytic hormone. Embryo is responsible for its own life by producing the sufficient level of the hCG. PMSG is a bigger molecule. It can not pass through kidney so can not pass through the urine and can not be detected. So blood is used to extract the PMSG. hCG will pass through kidney in the urine and can be separated from urine of the pregnant human female.
Clinical Use of FSH:

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Infertility due to failure of follicular growth It is used for superovulation Dose of FSH: Available preparations: FSH-P, Follitropin Follicular growth: 5 mg IM, 12 hourly for 2 days For superovulation: 5 mg IM, 12 hourly for 4-5 days
Clinical Use of PMSG:

Infertility due to failure of follicular growth It is used for superovulation Dose of PMSG: Available preparations: Gestyl (by Organon), Folligon , Fostim Follicular growth: 1000 IU, IM For superovulation: 2500-3000 IU, IM once in a cow
Clinical Use of LH:

For treatment of follicular cyst For treatment of delayed ovulation Dose of LH: Available preparation: Lutropin For delayed ovulation: 12.5 mg, IM For treatment of follicular cyst: 25 mg, IM
Clinical Use of hCG:

For treatment of follicular cyst For treatment of delayed ovulation Used in the Rig test Dose of hCG: Available preparations: Pregnyl (by Organon), IVF-C (by LG) For delayed ovulation: 5000 IU, IV For follicular cyst: 10,000 IU, IV
Rig Test
Purpose: To confirm the suspected cases of cryptorchidsm Action: hCG simulate the production of testosterone in blood Take blood sample (Sample A) 30 min before giving the injection of hCG. Then give a challenge dose of hCG i.e. 10,000 IU Collect the blood sample (Sample B) again 30 min after giving the injection. Compare the both samples (A& B) for the level of testosterone. Result: High rise in testosterone level in descendant testes but no difference is seen in cryptorchidsm. For testicular descent in pups: give a dose of 500 IU per pup, 2 times in a week for 4 week. ----------------------------------------------------------------------------------------------------------------
Delivered by: Dr. Maqbool Ahmad Repeat Breeding

When a female is not conceiving after 3 consecutive breeding without any clinically apparent signs of abnormality. It is the major cause of economic losses in dairy animals.
Causes:
There are four major sources; i) Female causes ii) Male causes iii) Owner causes iv) Doctor/veterinarian causes
Female causes of Repeat Breeding

i) Genetic problems: In case of white heifer disease; some parts or segments of reproductive tract are missing which form a blind pouch. In such cases, some time fallopian tube or horn (partially or completely) or cervix is missing. ii) Endometrial glands are missing: In the absence of endometrial glands, there will be not uterine milk result into early embryonic death. iii) Certain other developmental abnormalities of reproductive tract; like Double cervix: natural mating may be successful but A.I. may not. iv) Certain other problems affecting ovary: such as; Partial hypoplastic ovary v) If animal is not normal: chromosomal configuration is XXY, then there will be no chances of normal fertilization. Hormonal causes: vi) Follicle not ovulating: Following four factors are responsible for ovulating the follicle; i) LH surge ii) Intrafollicular fluid iii) Enzymatic thinning of wall of follicle iv) Smooth muscles present in follicle wall Insufficient level of LH, estrogen and P4 hormones also contribute in the failure of follicle ovulation. If LH surge not come, follicle will not go to rupture and it will persist and may lead to follicular cyst. In follicular cyst, animal will show irregular estrus. If receptors of LH are not responding, then ovulation also may not occur. vii) Delayed ovulation: It is very common in cross bred animals. Ova start to degenerate after 48 hours of start of estrus, either ovulation occur or not. This delayed ovulation may also occur in uterine infections due to which estrus duration is prolonged. Signs may be apparent or subclinical. Stress is one of the cause of ovulation. Stress by run to the inseminated animals also seen to enhance fertilization rate. viii) Any infection in the female genital tract; it may be specific or non-specific.
When zygote descends in uterine horn, then due to infection; rise in pH of uterine environment leads to the early embryonic death. ix) Salpingitis: It is responsible for development of ectopic pregnancy (mostly common in human females), in which zygote is unable to reach in the uterus due to inflammation of oviduct. Normally, parsitaltic and segmental contractions are responsible for transfer of ova and spermatozoa in ampula where fertilization has to take place. But in case of infection, these contractions are unable to produce and rolling of ova not occur. If by chance, fertilization occur then zygote is unable to survive in fallopian tube. x) Tube patency: refers to fallopian tube blockage. Fallopian tube blockage is due to any infection in F.T. Uterine TB is the most common problem of female women in which tube patency occurs. xi) Ovaro-bursal adhesion: If ovaro-bursal adhesion occurs, then fimbrial end unable to capture ova and ova may go into peritoneal cavity. This capacity is lost due to adhesion. xii) Environmental factors; causing early embryonic death xiii) Nutritional deficiency
Male Causes of Repeat Breeding

i) ii) iii) Testicular degeneration: It could due to many reasons as: Persistent high fever, Mumps, Chemical intoxication, Prolonged use of antibiotics. Crysptorchidsm: there may be one or both testes which not descend in scrotum. Any infection in male genital tract: It may be viral, bacterial or protozoal etc. Usually causative organism lodges in the accessory sex glands and ducts. Due to infection, fibrosis may also occurs of testicular parenchyma. Fibrotic Testes: Functioning will not be normal. Sperm concentration in the semen will be very low or no sperm seen in the semen. Genetic cause(s): such as small and fibrosed testes. Chromosomal abnormalities: either XXY or XYY Hypoplastic testes: give rise azospermia which is a genital disorder Unsuccessful mounting: weak thrust leads to partial ejaculation. Excessive use of bull Mishandling and poor quality of semen
iv) v) vi) vii) viii) ix) x)
Doctors/Technicians role in Repeat Breeding

As we know that more than 70% population of buffaloes show silent estrus. But this percentage is very low in case of cattle. Due to silent estrus, conception rate is very low due to some reasons: - inability to detect estrus at right time (estrus at proper time). - Estural behaviour of buffalo does not support the visual detection of estrus such as: buffalo in heat does not mount on others. i) Insamination during wrong time; in other phases of cycle instead of estrus period: In mid cycle, follicle and C.L. may be present. Due to estrogen production, animal show estural behaviour and may discharged mucus out. At this stage, animals may be insaminated but there will be no conception. ii) Improper handling of semen; Temperature for proper storage of fluid semen is 4C, which is maintained with ice packs. For frozen semen, liquid nitrogen ( temperature: 8
iii) iv)
v)
-196C) is used. It is very important to withdraw a semen straw from liquid nitrogen container with proper care and special care. Thawing is also very important in relation to time and temperature factor. After picking a straw from container: thawing is done at 37 C. 25% spermatozoa destroy in freezing and also same quantity in thawing. So a semen dose we get, has equal concentration of spermatozoa to that of fluid semen. Sperm conc. in a semen dose for cryopreservation: 20 million Sperm conc. in a fluid semen dose: 10 million Important step for preparing/Loading AI gun a) Lifting of goblet b) Picking of a semen straw c) Thawing of semen straw: During thawing, -60C to -15 C is very critical temperature which has to pass quickly. If you are thawing at 37 C, then dip the straw for just 30 sec to 1 min. If thawing is done in boiling water (100C) then just give a dip. So we conclude that temperature and time duration is very much important. Technicians mostly do thawing by rubbing of semen straw by taking it in the palms. In this way, there is no uniform thawing. Technicians do not dry the semen straw after picking from container and quickly load in the gun which should not be practiced. Remember that; a single drop of water coming along with wet straw is sufficient to spoil the semen dose. Proper loading and cutting of straw is also very necessary. Insamination must be completed within 5-10 min after thawing of semen. Technicians are unable to pass AI rod properly and insaminate at proper place. Detection of external oss of cervix is very important to pass AI rod successfully. Otherwise it may rupture the vaginal wall.
Owners role in Repeat Breeding

Proper detection of estrus is the first duty of owner. Owner must be vigilant to diagnose successful estrus. Heat signs: i) Ballowing ii) Restlessness iii) Off feed iv) Frequent urination v) Mounting behaviour (in cattle) vi) Stand to be mount (Standing heat) vii) Mucus discharge from vulva viii) Swollen, hyperemic and edematous vulva Internal signs: i) Vagina become glistening and pinkish ii) Cervix is opened iii) Uterine horn show marked tonicity; coiled and tonic iv) Graffian follicle on the ovary which is near to ovulate v) Rudimentary and conical corpus Luteum on the ovary Excessive use of bull also contributes in the repeat breeding because it depletes the storage of sperms and ejaculate contains very less amount of spermatozoa. One bull can be used maximum for 80 times in a breeding season. Breeding season: Cattle: Mid February to onward Buffalo: Mid Nov. to Mid Jan. ---------------------------------------------------------------------------------------------------------------
Delivered by: Dr. Ijaz Ahmad Anaestrus: Reproductive Failure in Female Animals
Fertility: It is an ability of individual to reproduce. Conception rate is more appropriate measure. Infertility: It refers to temporary loss of ability of individual to reproduce. It is a curable condition. It must be differentiated from the sterility. Sterility: It is defined as the permanent loss of ability of an individual to reproduce. It is an incurable condition. It may be congenital or environmental acquired. Estrus: Reoccuring period of sexual receptivity accompanied by ripening of one or more follicles and terminate with one or more ovum shedding.
Anestrus
It must be manifested by: - Absence of estrus - Lack of physiological heat signs - State of complete sexual inactivity
Types of Anestrus
There may be true anestrus or reported anestrus. It may be physiological or pathological state. It is a sign in a verity of conditions. True Anestrus: In this condition, ovaries will be plain and there will be no functional C.L. For treatment purpose, first provide good nutrition to improve the body condition of the animal. Give mineral supplements. If this nutrition plan fails to resume the cyclic activity, then go for hormonal therapy as last resort. Inject any one of the follitropin preapartion: FSH or FSH-like preparations, GnRH or PMSG for inducing cyclic activity by boosting follicular growth. As management tool, provide stress-free environment to the animals. Reported Anestrus: Animal is cyclic one and CL is also present on the ovary but reported as anestrus. First provide good nutrition to improve the body condition of the animal. Then, inject PGF2a for the lysis of CL so that ovarian activity can be resumed. Manual rupture of CL is not recommended.
Physiological Anestrus
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In physiological anestrus, it may be the following one: i) Prepubertal anestrus ii) Lactational anestrus iii) Gestational anestrus iv) Aging anestrus v) Anestrus due to stress 1. Aging anestrus: No stage is comparable to menopause but most farm animals are not maintained in old age. But anoestrus due to aging is also reported in animals which are maintained. Anoestrous during aging occurs due to some age-related changes. There is alteration in the relationship of hypothalamus pituitary gonadotropin axis and follicle releasing axis becomes refractory; becomes not responding to ovaries. There are two possibilities: Ovary may not responding to release of gonadotropin. The release of gonadotropin from pituitary is very less or insufficient. 2. Seasonal Anestrus: Seasonal anestrus is seen in seasonal breeders such as sheep, horses, cattle and buffalo. But its incidence and duration is variable. No cyclic changes in reproductive tract have been observed during anestrus period. Ovaries are found hard, no pre-ovulatory size follicle is there and usually CL may be absent. Extent of seasonal anoestrous varies with species, breed and physical environment. Cattle and buffalo are polyestrous animals but maximum no. of animals comes in heat during few months of the year. Peak breeding season in buffalo is September, October and in cattle; it is March, April. Seasonal anoestrus is more pronounced in sheep and mare which is associated with the photoperiod (tonic LH surge). Mare is long day breeder whereas sheep is short day breeder. Use of gonadotropins is disappointing. It is best managed by increasing photoperiod (day light). 3. Lactational Anestrus: After parturition, ovarian activity should resume within 30-35 days. Quiet ovulation occurs 2 weeks postpartum. Priming of CNS with progesterone is required for pronounced estrus symptoms. Whereas there are two occasions in the life of the animal in which quite ovulation occur (without any evident estrus signs): 1) Postpartum ovulation ii) First estrus ovulation of animal life Duration of cyclic activity and onset of estrus cycle is influenced by: - Season of calving - Level of milk production - No. of young ones being nest - Postpartum involution If animal calved when season is going to out, it will result to onset of anestrus period. In high yielding animals, cyclic activity is also stopped. Suckling activity release cortisol, which ultimately suppress the LH from anterior pituitary which has to cause final changes in mature follicle and then ovulate it. Prolactin is the major hormone of lactation which is implicated to suppress the release of gonadotropins; thus ultimately contribute in lactational anestrus. Malnutrition and calving in out of season or when there is a period of fodder scarcity also hampered the cyclic activity of animal.
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Mare come in heat 2 weeks postfoaling (called foaling heat or Dhamaan). Conception may occur but chances are very low. Some mares are nervy at that time and experience lactational anestrus. 4. Nutritional Anestrus: Energy plays significant role in the onset of cyclic activity. Animals on high energy plan usually attain puberty early. Some minerals (e.g. phosphorus, selenium) are more important. Deficiency of phosphorus result in delayed puberty. It causes ovarian dysfunction and also depresses the signs of estrus. Such animals show mild type of signs and of short duration. Ratio (P:Ca) when disturbed then animal will not show significant type of heat signs. In such animals, discharge of mucus (clear, transparent and string) is a good indication of estrus. 80% animals were in estrus which discharging mucus. When there is deficiency of one mineral, then it affects the absorption/assimilation of other mineral. Some vitamins (e.g. A,E) deficiencies leads to the irregular estrus cycle and finally result into anestrus. 5. Anestrus due to Stress Any type of stress (it may be environmental or nutritional) affects the onset of cyclicity and there may be failure of ovulation. Stress results in increased hypothalamus-pituitary-adrenal axis which inhibits the hypothalamus-pituitary-gonadal axis. In this condition, length of estrus cycle is increased but estrus period is decreased.
Failures of Follicular Development

Environmental - Season - Nutrition - Lactation Ovarian - Cystic ovarian Deg. - Hypoplastic ovary - Freemartin Uterine - Pyometra - Mummification - Pseudopregnancy
Gonadotropin Insufficiency
Failure of Follicular Development Anestrus
CL persistence
Factors affecting Follicular Development

Failure of follicle development ultimately leads to the anestrus. i) Environmental factors: These affects the release of gonadotropins due to which there is insufficiency of gonadotropins. a) Season b) Nutrition c) Lactation ii) Ovarian factors: results to the failure of follicular development.
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a) Cystic ovarian degeneration (COD) b) Hypoplastic ovary c) Freemartin iii) Uterine factors: There will be persistence of CL on ovary and high level of progesterone in the blood. a) Pyometra b) Mummification c) Pseudopregnancy Note: If suckling in sheep is stopped 2-3 weeks postlambing, then it will return back to estrus.
Certain other abnormalities which affect the onset of estrus

Specie Cattle Abnormality Anestrus Silent estrus or Subestrus Anestrus Anestrus Causes Pyometra, Mummification Lactation, Nutrition Heat stress Season, Lactation Lactation, Nutrition Mechanism Persistence of CL Release of gonadotropins is affected Endocrine system is affected Release of gonadotropins is affected Release of gonadotropins is affected
Sheep Mare
Abnormalities of ovaries and uterus which result in Anestrus

i) Cystic ovarian degeneration (COD) ii) Hypoplastic ovary iii) Freemartin
i) Cystic Ovarian Degeneration (COD)

There may be: a) Follicular cyst b) Luteal cyst c) Cystic corpora lutea
Follicular cyst
If a follicle of size 2.5 cm persist on ovary for a period of 7 to 10 days, then this will called follicular cyst. It will be a thick walled follicle. As follicle size increases, there will be increased amount of estrogen come out, due to which intense heat signs and prolonged estrus period is observed. Cause: It may be due to LH insufficiency (absence of LH surge) Irregular estrus cycle and prolonged signs of estrus of intense type (Nymphomenia) is the
Luteal cyst
In this type mature follicle is unable to ovulate but partial luteinization occurs to form a cyst. Thin rim of luteal tissue is formed around the un-ovulated ovarian structure. It leads to the anestrus, if it persists.
Cystic corpora lutea

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There is a central cavity in the developing corpus luteum. This cavity is quite wider due to which cystic corpora lutea covers about 70% of total ovarian tissue. Diagnosis: Female shows male like behaviour with certain physical changes; heavy neck and well muscles development, change in voice. Due to persistent high level of estrogen, there is relaxation of sacro-sciatic ligament and tail head is raised which is called sterility hump. Treatment: GnRH, LH and LH-like preparations When luteal tissue develops, then give PGF2a to resolve it and resume the ovarian activity. Important Note: Its occurrence is high in high yielding animals. Genetic predisposition is also of great consideration (daughters of affected mothers may prosper the problem).
ii) Hypoplastic ovaries:

It may be unilateral or bilateral. If unilateral, then there are chances to reproduce. Ovaries become hard, small and no preovulatory size follicle is found there. In bilateral hypoplastic ovaries, pelvis is not completely developed. Juvenile pelvis is small and animal is leggy one.
iii) Freemartin
In bovines, if female is born in co-twin with male, then she is always sterile. It is due to effects of male hormones on the female. Male gonads develop earlier in fetal life, hormones transfer from male to female side of same placenta which suppress the female reproductive tract activity. Hence, female is sterile by birth. Freemitis: A turbulence in the mid uterine artery is known as freemitis.
Prolonged Diestrus
It is unique in equine. Cause: Spontaneous prolonged life of cyclic CL It is the major cause of anestrus during breeding season. Why life of cyclic Cl is prolonged? It happens due to failure of release of PGF2a.
Disorders of Fertilization
1. Disorders of fertilization are discussed under: Fertilization Failure 2. Atypical Fertilization
1. Fertilization Failure
It may result from death of the egg or sperm before their fusion. There may be structural or functional abnormalities in egg or sperms. There may be some physical barrier in the fusion i.e. occlusion of the fallopian tube due to which transfer of gamete is affected. An-ovulation occurs in case of COD. No ovulation >>> No fertilization. Abnormalities of sperms 14
These contribute in the fertilization failure. These defects include: - structural defects of sperm (DNA defects) - Aging of sperm; due to which there is alteration in acrosomal cap - Enzyme leakage associated with storage of sperms - Fertility and integrity of acrosome are correlated. Fertilizable life must be differentiated from actual life of gamete. Fertilizable life is less than actual life. Abnormalities of egg Several morphological or structural abnormalities may be associated with: i) Damage to zona pellucida ii) Ovum may be giant size or it may be lentil shaped In such abnormalities, there will be no fertilization, but if it occur by chance then zygote may not further develop.
2. Atypical Fertilization
Several abnormalities are associated with atypical fertilization: i) Monospermy with two pro-nuclei ii) Polyspermy iii) Absence of pronucleus formation: it may be resulted by toxic medicine or exposure to injurious rays. iv) Aging of sperm/ovum, stress and unfavourable temperature also contribute in the fertilization failure.
Structural and Functional Causes of Fertilization Failure

Cause Abnormality Structural Abnormalities Congenital - Mesonephric - Cervix duplex - Unicornueus uterus Acquired - Tubal adhesion - Blockage of fallopian tube Functional Abnormalities Hormonal - COD - Changes in uterine secretion Managemental Specie affected Common in swine and also in cattle All species Mechanism Transport of sperm is affected Transportation of gamete is affected Ovulation is affected, an-ovulation and transport of gamete is affected. Results in death of egg or sperms No ovulation or anovulation
All species
- Delayed insamination All species - Early insamination - Wrong time insami. (Insami. in mid cycle)
Infectious Form of Infertility

Uterine environment is very hostile for growth of microorganisms. The temperature, pH and flora is very suitable for harboring an infection. Infection in the uterus ultimately results into: - Impair transport of sperms
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- Death of the spermatozoa If fertilization occur, then zygote has to descend down in the uterine horn, uterine environment will result into early embryonic mortality (early embryonic death). If early embryonic death not occur, then there may be still birth or delivery of a weak calf.
Specific Infection:
In specific infections, the predilection site of causative organism is reproductive tract, lodge there and proliferation occur. These do not require any predisposing factor. For example; brucellosis, leptospirosis, compylobacteriosis. Such infections ultimately cause infertility. Infections are enzootic in nature (large no. of animals in a herd is affected).
Non-specific Infection:
Infections caused by opportunistic pathogens. These require predisposing factor. These are sporadic in nature. How infection by Non-specific organisms is prevented? i) Physical barrier: It includes: - Cervical seal - Vulvular sphincter These barriers prevent the entry of organism into the reproductive tract. Any damage to these barriers may result into establishment of infection (i.e. cervicitis, vaginitis and endometritis etc.) ii) Natural defense mechanism: It is influenced by endocrine system. If life time, there are two occasion when these mechanisms are breached. i.e. Estrus and Parturition. During these both occasions, there is an increased blood supply to reproductive tract, due to which more defense cells protect from any infection. At the time of parturition, there is strong uterine contraction along with flow of mucus which washes out the whole tract completely. - The mucus discharge has a diluting effect against the infection. - Uterus under the influence of high level of estrogen is resistant to any infection, where as uterus under the influence of progesterone is prone to infection. ----------------------------------------------------------------------------------------------------------------
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Delivered by: Dr. Huma Jamil Infectious Forms of Infertility in Cattle and Buffalo
There are two types of infections: i) Specific Infections: These are enzootic types of infections. For example; brucellosis, vibriosis, trichomoniasis, leptospirosis, etc. ii) Non-specific Infections: Infections are caused by opportunistic pathogens which are commonly found on surface. For example; corynebacterium, salmonella, staphylococcus, streptococcus, and colliform (E.coli).
Non-Specific Infections
i) ii) iii) Puerperal metritis Endometritis Pyometra
i) Puerperal Metritis
It is the metritis which develops during the puerperal period. It develops few days after the parturition due to abnormal 1st and 2nd stage of labour. It is followed by uterine inertia, dystocia and retention of fetal membranes. Causative agents: Causative organisms of puerperal metritis are: - Corynebacterium - Streptococcus - Staphylococcus - Clostridium - E. coli Features: There is infection in the uterus and fetal membranes hang outside the vulva. These organisms colonize in the uterus and produce endotoxins. There is septicemia and toxemia due to proliferation of organism. General symptoms of septicemia are characterized by: increased body temperature, decreased milk production, diarrhea and gradual weight loss). Cervix is partially opened. Vulva and vagina are swollen. Reddish mucus discharged out. Complications: - Respiratory system disorders like pneumonia. Arched back; animal walk with stiff gait. - Abscess formation on the kidneys, liver and lungs. Treatment: i) First, remove the fetal membranes; manually or by hormonal therapy. Manual removal is difficult if vulva is hyperemic and uterus becomes friable. In hormonal therapy, use PGF2a @ 25 mg. ii) Administer oxytocin @ 50 IU, for expulsion of fetal membranes and fluid. Thing to remember for use of oxytocin is that fetal membranes should be passed through cervix or hang through the vulva. iii) Never give estradiol (esp. in the buffalo) it will cause strong contraction due to which chances of uterine rupture increased. iv) Uterine lavage with physiological saline solution @ 49 C. It will flush the obstructive material (if any) in the uterus. v) Intrauterine antibiotic infusions should be given. Uterine pessaries can also be used. Antibiotic sensitivity test is very necessary before selecting the drug of choice.
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Antibiotic Culture Sensitivity Test: culturing is done on the nutrient broth. Incubate @ 37 C for 24 hrs. After that, place different antibiotic disks in the plate having solidified agar. These disks may be of: Norfloxacin, Gentamicin, Kenamycin, Neomycin, Penicillin, Chlormaphenicol, Oxytetracycline. Then incubate for 24 hrs @ 37 C. Note the zone of inhibitaion. The more clearer and well developed zone around a disk indicates that drug is more sensitive and should be selected as drug of choice. Limitations of Culture Sensitivity test: Norfloxacin is not used intrauterine because it causes severe irritation. Others are non-irritant. It is very strange to know that invitro and invivo results are not similar. Other limitations are: Each organism may not give any growth on nutrient agar and in aerobic conditions such as clostridium; it must require anaerobic environment to survive. Limited time for culturing may not give appreciable growth of organism on the plate. Some organism such as Mycobacterium spp. require more time. Some fastidious type of bacteria such as Mycoplasma and Comyplobacter give no growth.
ii) Endometritis
It refers to the inflammation of endometrium. It is very common condition of cattle and buffalo. It usually develops after parturition. Clinical signs: Presence of pale yellow exudate with mucopurulent discharge from the vulva. Treatment: Best choice is administration of PGF2a @ 25 mg or 500 IU, I/M. By this treatment, animal will return to estrus within 4 to 5 days after flushing of infection.
iii) Pyometra
It refers to the accumulation of pus in the uterus. It usually develops after endometritis. It may be due to luteal cyst. It may develop after any specific infection such as vibriosis etc. It may be confused with pregnancy. Differential Points of Pyometra from Pregnancy: a) No fetal membranes b) Thin uterine wall b) No cotyledons d) No freemitis e) Corpus luteum is present on the ovary f) No conceptus DDx with Endometritis: - Corpus luteum is present in both conditions (pyometra and endometritis). - Uterus is enlarged in both cases. - In pyometra, uterus is filled with pus. While in endometritis, there is yellowish sticky discharge Treatment: First of all, regress the CL by administering PGF2a. For evacuation of uterus, use oxytocin @ 40-60 IU and stilboesterol @ 25-40 mg IM. Then go for parenteral administration of antibiotics.
Specific Infections
(i) Brucellosis
Syn: Bang disease Causative organism:
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B. abortis in Cattle and buffalo B. mallitensis in Sheep and Goat B. canis in Dog and cat B. suis in Swine It is gram ve bacteria which can cause abortion @ 6-8 mo of pregnancy in case of bovines. It can also cause still birth. Transmission: Mostly, by ingestion of contaminated food but may be through contact; nostrils, eyes and even through artificial insamination (AI). It does not transmit through natural mating because vulva acts as a physical barrier to infection. Brucellosis is more fatal for pregnant animals because infectious agent is more attractive towards fetus, placenta and lymphoid glands; because of more production of Erythritol which is a simple sugar. More erythritol production >> More invasion of bacteria >> Placentitis >> Abortion Aborted fetus and cow which abort the fetus are the major source of infection. Secretions from the aborted fetus, fluid and vaginal discharge are also full of brucella. Brucellosis infection may give rise to retention of fetal membranes which ultimately result into uterine inertia and puerperal metritis. Diagnosis: (i) Staining of material obtained from the aborted fetus, placenta and placental cotyledons. Prepare stain as: 20% carbol fuschin + 3% acetic acid + 10% Methylene blue Brucella will stain red and background will be black. It is first confirmatory test. (ii) Milk Ring Test (MRT) / Bang Ring Test: It is used as screening test. Antigen is prepared and then stained with methylene blue. Take 1 ml of milk in a test tube. Add 1 to 2 drops of stained antigen in the test tube. Mix it well by shaking. Incubate it for 1 hr @ 25-30 C. Result: If +ve: blue ring is formed on creamy surface of milk, whereas remaining is white. (iii) Rose Bangal Test (RBT) / Slide Agglutination Test (SAT) It is also a screening test. Antigen is prepared. Take 1 drop of serum on slide. Mix it with 1 drop of antigen. Result: If +ve then there will be formation of clumping and agglutination occurs. (iv) Complement Fixation Test (CFT) It is used as confirmatory test. Immunoglobulins (IgG and IgM) are detected. (v) ELISA : is also used as confirmatory test. Control: Vaccination: Attenuated vaccine having Brucella strain 19 is used in calves. Whereas killed vaccine having brucella strain 45/20 is used in heifers/cattle. No vaccination in bulls which are used for natural mating. If a bull semen is being used for AI, then it is necessary to perform screening test to declare the bull free from brucellosis infection.
(ii) Trichomoniasis
It is a venereal disease which is caused by Trichomonas fetus; a protozoa having an undulating membrane when observed microscopically. It usually causes; endometritis, pyometra, mucopurulent discharge, abortion and sterility in animals. Transmission: It occurs by coitus/service or through AI. In bulls, infectious agent colonizes in the crypts of prepuce and penis and these bulls are major cause of venereal transmission. In female animals, it multiplies in vagina and cervix within 3 weeks. In AI, as semen is directly pour into the body of uterus, so infection goes to the uterus and may cause pyometra. Importance of trichomoniasis:
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It got much importance because infection is associated with: i) Repeat breeding ii) Early embryonic death iii) Abortion (usually occurs in 2-4 mo of gestation but it may occur @7 mo of pregnancy) iv) Degeneration of fetus in the uterus, which ultimately leads to endometritis >>> PGs will not secrete and CL will persist on ovary v) Cow suffering from this infection develops agglutinating antibodies in its vaginal mucus and so after some time, auto recovery occurs as infection is washout. Diagnosis: In females: History of 60-90days non-return to estrus rate together with repeat breeding, along with purulent vaginal discharge, endometritis, pyometra and abortion (mostly in 2 to 4 mo of pregnancy). For direct demonstration of organism in vaginal mucus, collect the mucus from vagina by use of speculum and hand. First, lubricate well with oil or liquid paraffin, never use soap for this purpose (esp. in mare). Symptoms are quite similar to that of compylobacteriosis infection because irregular interestrus is observed in both cases. In males: Wash prepuce with physiological saline solution (10 ml). Take this solution in a test tube and centrifuge it. Prepare a warm slide to observe under microscope at 10X. It must be observed within 6 hrs after sample collection. Preputial scrapings mixed in physiological saline solution can also be used for diagnostic purpose microscopically. Aborted fetus can also be a best choice for diagnosis. Media used for transport of sample to diagnostic lab.:i) Buffer saline solution having fetal calf serum ii) Lactated Ringers solution iii) Dificomedia It can be cultured on: Oxoid media, and we can also use Dougles broth and Glucose serum broth for this purpose. Serological Test: Vaginal mucus agglutination test (VMAT) Treatment: If pyometra is there, then go for enucleation of CL, not manually but with PGF2a. Repeat it after 10-12 days. In cows, give sexual rest for 3-5 cycles in order to develop solid immunity and to increase fertility rate. In carrier bulls: use ointment 1% Bavaflavin (contains Trypaflavin+Surfen) on the prepuce after washing. Use dimetrinidazole @ 25 mg/kg b.wt. for 5 days orally or 75 mg/kg b.wt. TID I/V. Reinfection may occur in bulls so best choice is culling of carrier animals.
(iii) Compylobacteriosis
Syn: Formerly known as vibriosis It is a venereal bacterial disease which is caused by: Compylocater fetus; specie is C.fetus venerealis. Serotypes are A and B Compylocater fetus: specie is C.fetus fetus. Serotypes are A,B and C. Most imp is serotype C. Transmission: Through coitus/service or AI Symptoms: almost similar to that of previous one Infection exists in two forms:
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a) Acute: associated with causing infertility b) Chronic: It usually causes abortion. Acute infection: develops vaginitis, cloudy mucopurulent discharge, reddening of cervical mucosa, once uterus affected result into low grade inflammation which leads to infertility. Estrus cycle may be shortened or prolonged. Fetal membranes are severely affected and there is dark purulent material. Immunity develops after 2 to3 months. Ascending infection may involve the oviduct to causes salpingitis which is then characterized by sterility (permanent infertility). Bulls are symptomless carrier; their genitalia appear normal but their breeding records the prevalence of infection in a herd. Infected cows usually develop local antibodies i.e. IgA, IgM, IgG. Among these, IgG seen in uterus; and IgA & IgM in the vagina and cervix. These antibodies persists upto 10 mo in cervix and vagina. Recovered animals exhibit low fertility rate. Diagnosis: History and presenting complaints of repeat breeding, endometritis, pyometra, early embryonic death, abortion (usually @ 4 to 6 mo of pregnancy) and low fertility. Stain the organism which is observed as gram ve and looks like Flying Seagull, commashaped under microscope. Isolation of organism is done from genital secretions and semen of bulls. In bull, isolation may be done by aspiration method from prepuce and penis. Staining may be difficult; so go for FAT and IFT. For culturing, Bartlett technique is used. Vaginal mucus agglutination test (VMAT) may be used for diagnostic purpose. Media used for culturing of organism: i) Thyoglyconate media ii) Cooked meath broth iii) Veal infusion broth Transport media: Transport enrichment media; is used to store the organism for upto 2 days at normal room temperature. It contains solidified bovine serum with antibiotic. Treatment & Control: In male: Fat free cream containing 1% neomycin and polymyxin applied on the penis and prepuce after sedation. Streptomycin and erythromycin pastes can also be used. In female: Sexual rest is very important. PGF2a may be used to resolve the CL. Intrauterine antibiotic infusions of streptomycin and erythromycin (non-irritant) should be used.
Culture and Sensitivity Test

i) Collect the sample and then transport it to diagnostic lab immediately for sensitivity test. ii) Transport the sample in nutrient broth, at 37 C, may stored for upto 24 hrs. iii) Next day, transfer it on sensitivity agar; pH should be maintained. iv) Use different disks of antibiotics (i.e. tetracycline, gentamicin, penicillin, kenamycin, neomycin, norfloxacin) for sensitivity test. Ciprofloxacin is irritant, so it never use it. v) Then incubate it for 24 hrs at 37 C. vi) After incubation, observe for formation of zone of inhibition around each disk. Sterilization of glass ware is very necessary before test. A little residue or foreign material may change the pH of the media. Glass ware should be sterilized in hot air oven @ 170 C for 10 min. Media is sterilized in autoclave @ 121 C at 15 Pascal pressure.
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(iv) Leptospirosis
It is caused by a bacterial agent; Leptospira pomona. Clinically disease is characterized by: high fever, mastitis, and abortion in last trimester. Beside abortion, infection may be associated with retention of fetal membranes, metritis which leads to infertility. There is also hemoglobinurea because kidneys are severely affected. Udders become flaccid; thick yellow clotted or even some time blood mixed milk come out. Transmission: These are spirochetes which excrete in urine. Infection develops by ingestion of infected or contaminated food. It is no typical venereal disease, so no transmission through coitus or service has been observed. Diagnosis: History and presenting complaint of abortion associated with acute illness and blood-mixed milk On the basis of serology, antibodies may be drawn in the dam immediately after abortion. Before abortion, serum is collected from fetus for antibodies titer. But it is not possible in the fetus because it causes autolysis of fetus. Urine may be used for organism isolation. In serology; it is demonstrated that before abortion; serum titer is very high i.e. 1:1600 but it decreased to 1:400 after abortion. Transport Media: It may be 1% serum albumen and 5% flourosil Treatment: It is a self limiting disease. Strict quarantine should be done, if a new animal is going to introduce in an existing stock. Use streptomycin @ 25 mg/kg b.wt.
(v) Salmonellosis
It is a zoonotic problem which is caused by: Salmonella dublin and Salmonella typhimurium in cattle Salmonella abortis ovis in sheep Infection is characterized by: high fever due to toxemia, severe diarrhea or even dysentery. Due to toxemia, endometritis occur which causes the release of PGF2a ultimately leads to the lysis of CL, hence animal abort. Transmission: Ingestion of infected pasture and food. Since, organism is excreted in the feces and urine , so animal on grazing contaminate the pasture which becomes a source of infection for other healthy animals. Following abortion, uterus becomes severely affected. Animal may die to toxemia and dysentery. Otherwise animal excretes Salmonella organism for years in the feces. It is very important to dispose off the fetus properly, because it may act as a source of infection and cause zoonotic problem. Diagnosis: Signs and symptoms. Isolation of organism then go for staining (simple gram staining) and observe under microscope pink colored. Culture it on McConkey, brilliant green or SS agar. Serologically, you may go for ELISA and widal test (as used in human). Treatment: In past, chloramphenicol was used very successfully but now it is banned to use. Ciprofloxacin is the drug of choice most effective in salmonellosis. 22
(vi) Infection Mycoplasma

It is caused by bacteria; Mycoplasma spp. which is very fastidious type of bacteria; isolation is very difficult. It may cause abortion. Infection may be diagnosed on signs and symptoms. Alike isolation, culturing is also quite difficult as it requires more time i.e. 7-8 days for appreciable growth. Serological tests also helpful for diagnostic purpose.
(vii) Mycobacterium tuberculosis (TB) Infection

It does not cause abortion directly but affect the ovary, fallopian tube and uterus. Small nodules are formed which persist on the organs for long time result into loss of activity. Thus, it leads to low fertility and sterility (permanent fertility).
Specific Infections in Sheep

(i) Compylobacteriosis
Syn: Ovine vibriosis Causative organism: C.fetus intestinalis Transmission occurs through ingestion of contaminated food material. No venereal transmission. Organism resides in the gall bladder, intestine and placenta. Signs: Placentitis, necrosis of fetal cotyledons, abortion 3 days after fetal death. Diagnosis: Same isolation and identification as discussed previous. Serologically, IFT and FAT
(ii) Brucellosis
Causative organism: B. malletensis and B.ovis In male it causes epididmytis and abortion in female. Low fertility followed by abortion.
(iii) Lepstospirosis
Sheep is quite resistant to this infection.
(iv) Salmonellosis
Causative agent: S.abortus ovis It causes abortion in last trimester. Other findings are: placentitis and fetal bacteremia. Diagnosis: Signs and symptoms. Isolation and culturing of organism Treatment: In past, chloramphenicol was used successfully but now it is banned. Ciprofloxacin is the drug of choice.
Viral Infections
(i) Infectious bovine rhinotracheitis (IBR)
Syn: Infectious pustular vulvovaginitis Infection occurs in two forms: i) Respiratory form ii) Reproductive form; cause infection of vulva and vagina. Ulcers develop on mucus membrane of vulva and vagina. Infection may goes to uterus due to which yellowish pus comes out. Abortion occurs during 4 to 7 mo of pregnancy. Transmission: sexually (service and AI) and through inhalation. 23
Diagnosis: Signs and symptoms and based on serology. Treatment: No specific treatment. Supporting therapy and antibiotic therapy to avoid any secondary infection.
(ii) Bovine Viral Diarrhea

Syn: it is also called mucosal disease It is a mild, acute to chronic viral infection. Virus infect the pregnant cow, fetus may die or born with cerebral hyperplasia. Diagnosis: Signs and symptoms are diagnostic and perform serology. Treatment: No specific treatment
Other Viral Causes of Infertility:

Parainfluenza-III (PI-3) virus Epizootic bovine abortion Blue tongue Specific bovine venereal epididimytis () vaginitis () ---------------------------------------------------------------------------------------------------------------
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Delivered by: Dr. Shujait Ali Diagnosis and Therapeutic Management of Post-partum Infections in Bovines
Relevances: Uterine infections are a major cause of economic loss to the cattle industry. Incidence of uterine infections are influenced by calving management, general sanitation, pathogenic organisms, endocrine factors, lactation, nutrition and other environmental stress factors.
Etiology, definition and classification of post-partum infections:

Etiology: The uterus is normally protected from bacterial contamination by the vulva, vestibular sphincter and cervix. During and immediately after parturition, these mechanical barriers are breached and the uterus is normally contaminated by a variety of pathogenic and non-pathogenic microorganisms. Most of these bacteria are only transient residents and are promptly eliminated by the uterine defence mechanism during the puerperium. In some cases, however, pathogens persist in the uterus and cause disease. The organism most commonly associated with uterine disease in cattle is Actinomyces pyogenes. The gram-negative aeroboes Fusobacetrium necrophorum and Bacteroides melaninogeneicus are frequently associated with A. pyogenes. Bacteriodes decrease chemotaxis and inhibits phagocytosis by neutrophils allowing A.pyogenes to persist. A variety of other micro organisms are occasionally associated with uterine disease in cows and include Coliform, Pseudomonas aeuroginosa, Staphylococci, hemolytic Streptococci and others. Clostridium spp. occasionally infect the uterus and cause severe gangrenous metritis. Some of the organisms that transiently contaminate the uterus during the post-partum period produce penicillinase; this should be a considered during the selection of drugs and routes of administration. In cows, with a normal puerperium, the uterus is nearly free of bacterial contamination by 4 weeks after calving. Major groups of uterine bacteria: Coliform bacteria E. coli Proteus spp. Enterobacter spp. Incidental bacteria:
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Streptococci spp. Staphylococci spp. Pasteurella spp. Bacillus spp. Corynebacteria:
C.pyogenes (now called Actinomyces pyogenes) Gram negative anaerobic bacteria Bacteroids spp. Fusobacterium spp. Gram positive anaerobic bacteria Clostridium perfringes C. sporogenes Other Clostridium spp. INTERESTING FACTS C. pyogenes, Fusobacterium and Bacteroides spp. act synergistically to enhance the diseases. C. pyogenes is the most significant pathogen of the intermediate and post ovulatory periods. Many types of bacteria intermittently inhabit the uterus in the post partum period but most have little effect on fertility. However, they can give protection to the C.pyogenes, Fusobacterium and Bacteroid from the antibiotics because some of them secrete penicillinase which destroy the sensitive penicillins and cephalosporins. Definitions: Definition of uterine infections have considered character of uterine discharge, day postpartum, clinical findings and endocrine status. Unfortunately, clinicians and researchers have vaguely applied terms such as metritis and endometritus when describing uterine infections, which has contribute to confusion among veterinarians in the definition and economic impact of uterine infections. Therefore, specific definitions of related entities are presented as follows. Postpartum period: It is defined as the period from parturition to complete uterine involution. Metritis: Metritis is a result of severe inflammation involving all layers of the uterus (endometrial mucosa and submucosa, muscularis and serosa). Endometritis: Endometritis is characterized by inflammation of the endometrium extending no deeper than the stratum spongiosum. Pyometra: Pyometra is characterized by accumulation of purulent exudates of variable amount within the endometrial cavity, persistence of a corpus luteum and suspension of the estrous cycle.
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Classification:
(a) Puerperal period: It begins at the time of calving and continues until the pituitary gland becomes sensitive to GnRH at 7 to 14 days postpartum. (b) Intermediate period: It begins with increased pituitary sensitivity to GnRH and continues until the first postpartum ovulation. (c) Post ovulatory period: It begins at the time of first ovulation and last until involution is complete. It is about 45 days postpartum in normal cows. Puerperal period: During the puerperal period, mixed population of bacteria remains present in the uterus. Their numbers increase for several days, then start to decrease as involution progresses in normal cow. RFM is a common problem of the early postpartum period which increases the risk of uterine infections. Establishment of C. pyogenes, Fusobacterium necrphorum and Bacteroides spp. occur in this period, if condition is favourable. Eventually these get localize and cause chronic type of metritis in intermediate and postovulatory period. Coliform and incidental bacteria are decreased in number or eliminated from the uterus during the puerperal period especially in cows that develop more persistent infection with C. pyogenes and Gram-negative anaerobes. The Coliform and incidental bacteria play no significant role in causing chronic metritis although in the early stages they may be associated with septic or toxic metritis. Life threatening infection occurs almost exclusively during this period like septic puerperal metritis. Many micro-organisms that cause septic puerperal metritis are susceptible to penicillin, so it is the antibiotic of choice for systemic treatment of the affected cows. In cows with puerperal metritis without systemic involvement, local therapy with tetracycline alone is usually adequate. Oxytetracycline is the antibiotic of choice for intrauterine therapy in the puerperal period if there is no septic infection (See the tetracyclines to know the reasons). Intermediate period: In the intermediate period, bacterial populations are reduced in the uterus of normal cows. But in normal cow, uterine infection gets localized and causes endometritis or metritis and purulent discharge comes out when the cow lies down. Intrauterine infusion of 1-2 gm of tetracycline in 20 to 40 ml of sterile water or physiological saline solution indicated because the uterus of many cows still contains mixed population of bacteria that may produce penicillinase. This therapy should be continued for a minimum of 3 days. Post-ovulatory period: Most common diseases occurring in this period are metritis and pyometra. The bacteria that seem to be associated with chronic metritis are C. pyogenes and Gramnegative anaerobic bacteria (Fusobacterium necrphorum and Bacteroid spp.)
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These bacteria stimulate exudation of large members of leukocytes resulting in purulent exudates. Since most metritis of this period is the result of penicillin-sensitive micro-organisms (C. pyogenes and Gram-negative anaerobes) because most of the other bacteria have been eliminated up to this period. So, penicillin is the antibiotic of choice for the treatment in the post-ovulatory period. Intrauterine infusion of 10 lakhs IU of penicillin provides therapeutic levels and entire wall of the uterus for a minimum of 24 hours. Normal healing of the endometrium after elimination of C. pyogenes requires about a month. Therefore a cow should not be inseminated in next one or two estrous periods. Pyometra should be treated with estrogen / PGF2 and antibiotics (Penicillin is the antibiotic of choice).
INTERESTING FACT Inflammation of the oviduct, ovaries and ovarian bursa due to ascending infection from the uterus may be resolved and fertility may be restored in many cows but permanent functional impairment of oviductal epithelium may prevent successful fertilization and ova transport and the cow may become sterile. Treatment: Therapy of uterine infection has fallen into the broad categories of intrauterine therapy (antibiotics and antiseptic chemicals), systemic antibiotics and supportive therapy, and hormonal therapy. Intrauterine Therapy: A variety of antibiotics and antiseptic chemicals have been infused into the uterus of cows in attempts to treat postpartum infections. The bovine uterus is an anaerobic environment; thus antibiotics chosen for intrauterine use must be active in the absence of oxygen. In addition, most antibiotics and chemicals depress activity of uterine neutrophils and interfere with the uterine defence mechanism; therefore, the potential benefit of their use must be carefully weighed against their deleterious effects. Organisms that cause postpartum uterine infections usually are sensitive to penicillin, but bacterial contaminants during the first several weeks after calving produce penicillinase, which renders the drug ineffective if applied locally. By 30 days postpartum these organisms usually are eliminated, and intrauterine treatment with penicillin is more likely to be effective after that time The daily intrauterine dose of penicillin recommended to reach the minimal inhibitory concentration (MIC) for A. pyogenes is 1 x 106 IU. Oxytetracycline is commonly recommended for intrauterine therapy for postpartum infections. In a recent study, however, most isolates of A. pyogenes recovered from the uterus of cows were resistant to oxytetracycline and intrauterine treatment with large doses did not affect the frequency of A. pyogenes isolation. Furthermore, many preparations of oxytetracycline are irritating and cause chemical endometritis. If oxytetracycline is selected for intrauterine therapy, doses of 4 to 6 g/day have been recommended in one study.
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Intrauterine therapy with iodine solutions for the treatment of uterine infections is not recommended in all recent studies. Systemic Antibiotics and Supportive Therapy: A variety of broad spectrum antibiotics have been recommended for parenteral administration to cows with uterine infections. Penicillin or one of its synthetic analogues is most commonly recommended (20,000 to 30,000 IU / kg bid). Oxytetracycline probably is not a good choice for systemic administration because of difficulty in reaching the MIC required for A. pyogenes in the lumen of the uterus. Ceftiofur is a third-generation cephalosporin that has broad-spectrum activity against both Gram-positive and Gram-negative bacteria has been founded useful in the treatment of metritis. Moreover, ceftiofur has been reported to reach all layers of the uterus without voilative residues in milk. Subcutaneous or intramuscular administration of ceftiofur at a dose 1 mg/kg in dairy cows after parturition result sufficient concentration of ceftiofur and its active metabolites in plasma, uterine tissues and lochial fluid that exceedingly reported MIC values for common pathogens involved in metritis. Ceftiofur administered at a dosage of 2.2 mg/kg, daily for 5 days was efficacious in the treatment of metritis (rectal temperature > 103.1 F with foetid vaginal discharge). If dehydration complicates metritis, appropriate fluid therapy should be instituted and may be life-saving. Nonsteroidal anti-inflammatory drugs such as flunixin meglumine are used to combat toxaemia and improve appetite. Furthermore, cows with metritis may experience depressed appetite, affecting calcium and energy status. Consequently, therapy with calcium and energy supplements may be warranted. Hormonal Therapy: A variety of hormones have been administered to cows in attempts to prevent or treat postpartum uterine infections. Oestrogen has been administered to initiate or strengthen Myometrial concentrations, but its use is controversial. Contraction induced by oestrogen has been blamed for forcing the septic contents of the uterus not only through the cervix but also into the uterine tubes resulting in severe bilateral salpingitis. Uterine Defence Mechanism Normally, postpartum uterus has a heavy load of bacteria initially but these are eliminated by uterine defence mechanism in a normal cow. Phagocytosis of micro-organisms by neutrophils is so important bovine uterine defence mechanism. This defence system is stimulated about 2 days after parturition by the invading micro-organisms. Under pathological conditions, such as delivery of dead or weak calves, phagocytosis can be depressed for several days to several weeks. Trauma to the genital tract by removal of RFM and obstetrical procedures also depress phagocytosis.
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Possibly a number of antibiotics commonly placed in the uterus might also depress phagocytosis because of irritation. Uterine defence mechanism is stimulated by oestrogen and inhibited by progesterone. Oxytocin causes contraction of the myometrium if the organ is dominated by oestrogen. Thus, oxytocin is expected to be effective in aiding uterine evacuation if administered within 48 to 72 hours after calving. Doses of 20 to 40 IU repeated every 3 to 6 hours are commonly used. Likewise cows affected with dystocia, RFM, both and treated with PGF2a in early post partum, followed by a second treatment of PGF2a 14 days later, experienced a higher conception rate to first service than untreated cows experiencing a normal or abnormal parturition. Prostaglandin is the drug of choice for therapy of pyometra. The evacuation of the uterus in 85% to 90% of treated cows occurs in 3 to 9 days after treatment but the endometrial lesions should be allowed 30 days to heal before A.I. Prognosis: The prognosis for recovery from postpartum uterine infections varies with severity of the condition. Most cows with uncomplicated endometritis can be expected to recover. Metritis complicated by septicemia may result in permanent impairment of fertility, decreased milk yield, laminitis, or in extreme cases, death of the patient despite aggressive treatment. Pyometra is rarely accompanied by abnormal clinical signs other than anoestrus in cows and rarely endangers the health or life of the animal. Most cows recover promptly from pyometra, if the condition is diagnosed and treated early in its course. ----------------------------------------------------------------------------------------------------------------
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Delivered by: Dr. Nazir Ahmad

Diagnostic Ultrasound
Ultrasound: when we talk about ultrasound, then we must know what is a wave and its component. Since, ultra sounds propagate in wave. Wave: To describe a wave, two, three associated parameters must know to us; frequency, time period and wave length. Frequency: It can be defined as number of cycles completed by wave in one second. It is denoted with f which is equal to cycles per second (cycles/second). Unit is Hertz (Hz). Time period: It is reciprocal to frequency (T 1/f) . If refers to time required by a wave to complete one cycle. It is denoted with T and equal to 1/f. Unit is s (micro second) Wavelength: It is defined as distance covered by a wave in one cycle. It is denoted with . Unit is mm (millimeter). Sound waves: The waves of sound which are hearable by human ear, known as sound waves. The frequency of these waves ranges from 20 to 20,000 Hz. Subsound or Infrasound waves: The waves of sound whose frequency is less than 20 Hz (<20 Hz); ultimately not hearable by human ear are known as subsound or infrasound waves. Ultrasound waves: The waves of sound whose frequency is above than 20,000 (>20,000 Hz); not hearable by human ear are known as ultrasound waves. Diagnostic Ultrasound: wave frequency used in diagnostic ultrasound ranges from 1 to 10 MHz. (MHz = 106 Hz). Probes of 2.0, 3.5, 5.0, 7.5 and 10.0 MHz frequency are available in the market. Ultrasound: Ultrasound (also called sonography) is a diagnostic medical procedure that uses high-frequency sound waves to produce dynamic visual images of organs, tissues or blood flow inside the body. Principle on which ultrasound works is just opposite to that of radiography. Here reflected waves are important to make an image on the screen while in the radiography; absorbed waves leave a print on x-ray film.
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Parts of ultrasound machine

There are three parts; i) Transducer or probe ii) Main processor or monitor iii) Control panel / key board Transducer or Probe An ultrasonic transducer is a device which converts the energy into ultrasound waves. It is made up of unique type of crystal called piezo-electric crystals. Piezo is a Latin word which means; pressure. When voltage is provided to the device, sound waves are produced which enter into the body tissue. Fate of waves produced by transducer: There are three possibilities; either these waves reflected, refracted or absorbed completely by the body tissue. Hence, three phenomenon can be observed; i) Reflection ii) Refraction iii) Absorption or scattering Reflected waves are received back by the transducer, which then goes to processor and displayed on the monitor screen. Absorbed waves are not known to us because nothing goes back to the processor and displayed on screen.
Factors which determine the fate of waves

Factors which determine that how many waves will pass through reflection, refraction and absorption phenomenon. Nature of the tissue: More softer tissue >>> More waves will pass through and a very few will reflected back. More hard and compact tissue >>> Most waves reflected back and only a few absorbed / pass through the structure. Hence, image of the tissue or structure will be defined by its nature and consistency.
Advantages of ultrasonography
Here few advantages of ultrasonography are enlisted on the other diagnostic techniques i.e. radiography etc. (i) This technique is non-invasive (no surgery, no cut, no damage to the tissue by anyway). (ii) Image which is obtained is live and dynamic such image is known as real time image. (iii) This technique is innocuous (harmless). There is no side effects recorded in human beings. For this purpose, a scientist takes semen in a test tube. He attached probe and passed ultrasound waves through the semen. He observed that there is no effect on the quality (motility and concentration) of spermatozoa. (iv) This technique is used as a part of physiotherapy to treat certain chronic fractures. As transducer converts energy into sound waves which are helpful to heat up the affected area in order to enhance the blood circulation. Blood engorgement in a chronic fracture and injury carry the healing process. (v) Image obtained on the screen can be recorded on CD, DVD or computer memory for future use. While it is not possible in other techniques such as radiography. (vi) By use of this technique, it is possible to determine nature, size and location of the lesion (tumor, cyst or abscess). It will help us in surgical intervention thereafter. (vii) Image of the ultrasound can be printed for future use.
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Disadvantages of ultrasonography
(i) This technique can not be used for the examination of organs which are covered with bone; if organs are lying underneath or below the bone. For example, if we do ultrasonography of brain then it is not possible to observe the image of brain parts because a hard covering of brain in the form of skull does not permit us to view underlying tissue. (ii) Organs which are filled with air, they can not examine through ultrasonography i.e. lungs. In the lungs, most of the sound waves will scattered and not reflected back to give any image. (iii) Organs which are covered with fat, they can create some problem. It can be minimized by taking the image from that angle in which minimum fat come in the path. (iv) You should know normal appearance of the tissue/structure, then you can differentiate it with the abnormal. Additionally you should also know about the anatomy of other organs located around. Important Note: (i) For ultrasonography, first prepare the animal by clipping the hairs and wool from the site. Then use a gel before placing the transducer; it is applied to ensure a close contact with the body. This gel will eliminate the air between probe and body surface. Air between probe and body surface will damage the quality of image, and sometime it is possible that no image may obtained due to this defect. (ii) For pregnancy diagnosis, in large animals (mare etc) probe is placed in the rectum. False positive result may be accompanied with the true image as bladder is lying just under the uterus. (iii) Never depend on ultrasound report completely. It should be taken as additional aid for any diagnosis.
Display Formats
There are three formats of display in ultrasonography; (i) Amplitude mode (A-mode) (ii) Bright mode (B-mode) (iii) Motion mode (M-mode) (i) Amplitude mode (A-mode): It is an old version of display format. In this format, reflected waves are shown on the screen in the form of peaks and spirals. reflected waves --- peak height Increased peak height correspond the increased intensity of reflected sound waves. peak height indicates more hard structure; hence it tells use about the nature of tissue. It also tells about the distance of tissue from the probe. It is called one dimensional display. This display mode is not commonly used but some uses are there in human ophthalmology. Brightness mode (B-mode): It is most commonly used both in human and veterinary diagnostic medicine. In this format, reflected waves are shown by a series of spots on the screen.
(ii)
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Fluid filled structure such as graffian follicle will give a black image on the screen, whereas a hard bony, calcified structure will give bright image. Other soft images will give an image of contrast between black and white. e.g. Liver ultrasonograph is more black because there is more fluid in comparison to testical one, which is a mixture of black and white. B-mode is a two-dimensional display; you can assess that structure is how much deep from the surface and away from the sides. Images of abscess will be like pyometra but there will be a bright outline due to thick demarcation of abscess. Size of abscess can be determined with the help of scaling on the display. B-mode can detect: nature, location and exact size of the lesion (iii) Motion mode (M-mode): It is also very commonly used for examination of moving organs i.e. Heart; hence it is also called echocardiography. Heart function, intestinal movement is monitored through this display mode.
Selection of Frequency
Commonly used frequency for ultrasonography ranges from 1 to 10 MHz. Commercially, probes of different frequencies are available as follows: 2 MHz, 3.5 MHz, 5 MHz, 7.5 MHz, and 10 MHz
Imaging depth
It negatively (or inversely) correlates with the frequency of the probe. As frequency of probe is increased penetration power is decreased (less imaging depth) and vice versa. Optimum Imaging depth that is achieved by different frequencies: 2.0 MHz 30 cm 3.5 MHz 18 cm 5.0 MHz 12 cm 7.5 MHz 8 cm 10.0 MHz 6 cm
Imaging quality
It positively correlates with the frequency. As frequency is increased, better imaging quality is obtained. High frequency ---- Good image. Low frequency ----- Poor image Hence, frequency of probe set according to the depth of the structure. If you are going to examine deep structures (pregnancy diagnosis; fetus) then use low frequency probe. For superficial structure, use probes of higher frequency. In most of the ultrasound machines, probe of a single frequency is fixed but in modern era, such probes are also available in the market which are of dual frequency and you can switch on another frequency after switching off the first one. As a whole, it is concluded that probe of frequency 5.0 MHz is the best one for taking both superficial and deep images and it will be cost effective.
Types of Transducer
Transducer/probe used in ultrasound machine are of three types as follows; (i) Linear array (ii) Sector array
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(iii) Cortex array
(i) Linear array:

In this type, crystals (10 to 20 in number) are arranged in a row or line. Source of sound waves is different crystal. Image that is obtained will be rectangular in outline; because waves run parallel to each other.
(ii) Sector array:

There is only one crystal. When AC is provided then crystal starts rotating, due to which subsequent waves produced will be slightly in other direction. Source for different waves is same. As sound waves goes deeper, these will expand and cover more area. Hence, image will be wedge shaped. Merit: in comparison with Linear array: In linear array, when a tissue is scanned through a narrow space, then only waves propagate in a straight direction and scanned the structure come in their path (see figure). But in case of sector array, when probe is attached on such space, due to property of expanding the waves, nearby tissue is also scanned which is lying a bit on sides. Hence, scanning or imaging of more tissue area is possible with sector array transducer. Demerit: In linear array, it is easy to scan the whole tissue which is under the probe at a time while in case of sector array, due to such configuration of probe, it is not possible to scan the uppermost top sided tissue by placing the probe once on the tissue surface.
(iii) Convex array:

It is made up by combing the properties of both linear and sector array. There are many crystals which are placed in a convex fashion; to send the waves in multi direction (at different angles). Image will be just like that of sector array but it will be concave at the top.
Application of probes:
Linear and convex array probes are used transabdominally; placed topically to take the image of different visceral organs of abdomen. Linear array probes of different shapes are available; particularly to use transrectum; per rectum ultrasonography.
Echogenicity
Brightness in an ultrasound image/print is known as echogenicity. Fluid filled cavity/structure ---- echogenicity decrease Chronic infection (i.e. liver cirrhosis, epididymitis, calcified stone), echogenicity increase Fatty change in the liver (lipoma) --- echogenicity increase
Ultrasound Artifacts
Ultrasound artifacts are of three types: a) Acoustic shadowing: It is associated with highly reflected organ like bone, or stone in the kidney. The area which is just under that highly hard reflected structure, will look black than the normal on the screen. While there will be no evidence of fluid filled structure at that site. This artifact also seen in image of abscesses which are of curved border. E.g. Thickness of the abscess is same i.e. 2 cm but it will seem to be more at curved border in the image. This artifact is quite common. b) Enhanced through transmission:
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It is associated with poorly or weakly reflected tissue e.g. follicular cyst. Waves accumulated more at the area A than B and C (see fig) ; because waves not reflected back Hence, area under poorly reflected organ/structure is more brighter than normal. (iii) Reverbration: It is associated with a very small distance between probe and highly reflected organ. If highly reflected organ is lying just underneath the probe, then waves will bounce back by following reflection mechanism and will give no image. Instead of a clear image, we get multiple lines on the screen which reduce in length as propagate below. (iv) Mirror image: When you examine abdominal visceral organs; then diaphragm acts as a mirror. Image of same nature, size and at same distance is obtained on opposite side of the real image due to mirror imaging artifact.
Description of an Image
Echoic: This word is used for a very bright area seen in an ultrasonograph. Anechoic: It is used to describe a black area seen in an image. Diffused: If the entire area is a mixture of black and bright then; it is said to be diffused. To differentiate abnormal area from normal, terms hyperechoic and hypoechoic are used. Hyperechoic: echogenicity is more than the normal. Hypoechoic: echogenicity is less than the normal. Hence, in remarks we write that this much area seen hyperechoic at that site on image. ---------------------------------------------------------------------------------------------------------------End of Class Lectures (AR 405)
Remember: Only Portion of Dr.Nafees Akhtar is missing in this handout. Please consult the given handouts for all topics covered by said teacher. These handouts are available at photostat shop on your request. Thanks !!!
Compiled and Presented by:
Muhammad Sajjad Hussain DVM 8th semester For your suggestions and feedback: Email: dvmdoctors@gmail.com Contact: +92 322 6272 278 Visit: www.dvmdocs.webs.com Footnote: Compiler will take no responsibility, if any unintentional mistake found by reader. It is suggested to consult these handouts with your own note downed lectures. If any error and omission is found; then inform please.
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REPRODUCTION CLINIC - II

To the Point Class Lectures

FINAL EXAM NOTES COMPLETE*

MUHAMMAD SAJJAD HUSSAIN

Best of Luck !!!

Up to Date: 25 May 2011 W ednesday

Delivered by: Dr. Najib Ur Rehman Clinical Use of Reproductive Hormones

GnRH (Gonadotropin Releasing Hormone)

Factors which STIMULATE GnRH secretions

Factors which SUPPRESS GnRH secretions:

Clinical Uses of GnRH

Natural GnRH Preparations

Synthetic GnRH Preparations

Comparative Features of pituitary and placental gonadotropins

Clinical Use of FSH:

Clinical Use of PMSG:

Clinical Use of LH:

Clinical Use of hCG:

Delivered by: Dr. Maqbool Ahmad Repeat Breeding

Female causes of Repeat Breeding

Male Causes of Repeat Breeding

iv) v) vi) vii) viii) ix) x)

Doctors/Technicians role in Repeat Breeding

Owners role in Repeat Breeding

Failures of Follicular Development

Failure of Follicular Development Anestrus

Factors affecting Follicular Development

Certain other abnormalities which affect the onset of estrus

Abnormalities of ovaries and uterus which result in Anestrus

i) Cystic Ovarian Degeneration (COD)

Cystic corpora lutea

ii) Hypoplastic ovaries:

Structural and Functional Causes of Fertilization Failure

Infectious Form of Infertility

Culture and Sensitivity Test

(vi) Infection Mycoplasma

(vii) Mycobacterium tuberculosis (TB) Infection

Specific Infections in Sheep

(ii) Bovine Viral Diarrhea

Other Viral Causes of Infertility:

Etiology, definition and classification of post-partum infections:

Streptococci spp. Staphylococci spp. Pasteurella spp. Bacillus spp. Corynebacteria:

Delivered by: Dr. Nazir Ahmad

Parts of ultrasound machine

Factors which determine the fate of waves

(iii) Cortex array

(i) Linear array:

(ii) Sector array:

(iii) Convex array:

Compiled and Presented by:

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