ANSC 3401 Reproductive Physiology Lab Manual
Female Anatomy
Introduction: The structures of the female anatomy will be covered from the external
genitalia to the internal reproductive organs. The changes during the different stages of the
estrous cycle and pregnancy will also be covered.
Vulva: the outermost portion of the female reproductive tract
Made of two portions:
Labia majora – outer lips
Labia minora – inner lips
Proestrus:
Estrus:
Metestrus:
Diestrus:
Anestrus:
Pregnancy:
Vestibule: the common pathway of the reproductive and urinary tract
Terminal portion of the vagina
Site of the urethral opening (suburethral diverticulum)
http://www.ttuhsc.edu/som/obgyn/faculty/Prien...0Reproductive%20Physiology%20Lab%20Manual.htm (1 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Clitoris: erectile tissue located just anterior (inside) the vulva
Homologous to the glans penis in the male
Contains numerous sensory nerves and increases sexual excitement
in the female
during copulation
Estrus:
Vagina: copulatory organ and birth canal
Site of sperm deposition in most species
Contains many mucosal glands
Made of highly elastic muscle and connective tissue
Vaginal Fornix: blind pouches on either side of the cervix
Proestrus:
Estrus:
Metestrus:
Diestrus:
Anestrus:
Pregnancy:
Cervix: thick-walled, inelastic "tube" that separates the vagina the uterus
Usually a single structure except in species with duplex or didelphic
http://www.ttuhsc.edu/som/obgyn/faculty/Prien...0Reproductive%20Physiology%20Lab%20Manual.htm (2 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
uteruses
Made connective tissue
Prevents contamination of the uterus
Serves as a sperm reservoir
Produces cervical mucous
Shape is species specific:
Annular rings in cow and ewe
Corkscrew in pigs
Mucosal folds in mares
Os Cervix is the caudal opening (vaginal side)
Os Uteri is the anterior opening (uterine side)
Proestrus:
Estrus:
Metestrus:
Diestrus:
Anestrus:
Pregnancy:
Uterus: organ that contains the fetus during pregnancy
Produces hormones
http://www.ttuhsc.edu/som/obgyn/faculty/Prien...0Reproductive%20Physiology%20Lab%20Manual.htm (3 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Site of placental attachment
There are several types of uteri:
Bicornuate: two horns, no body, one cervix, one
vagina
Cattle, sheep, goats, swine and dogs
Biparite: two horns, one body, one cervix, one vagina
Horses
Duplex: two uteri, two cervix, one vagina
Rabbits and small rodents
Simplex: no horns, one body, one cervix, one vagina
Primates, including humans
Didelphic: two uteri (each with one body and one
horn), two vagina, two
vulva openings (Males of these species have a forked
penis.)
Kangaroos, opossums, and other
marsupials
Proestrus:
Estrus:
Metestrus:
Diestrus:
Anestrus:
Pregnancy:
http://www.ttuhsc.edu/som/obgyn/faculty/Prien...0Reproductive%20Physiology%20Lab%20Manual.htm (4 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Oviducts: paired structures that connect the ovaries and uterus
Also known as fallopian or uterine tubes
Transports ova and sperm
Made up of several segments:
Fimbria: finger-like projections that surround the ovary
to "catch" the oocyte
when released during ovulation
Infundibulum: funnel-shaped portion of the tube
connected to the fimbria
Ampulla-Isthmus Junction: site of fertilization
Uterotubal Junction: connection site of the oviduct
and uterus
Ovaries: paired structures that are the primary sex organs in the female
Produces the female gametes (oocytes)
Produces hormones
Follicles: contain the oocyte and liquor folliculi
Goes through several stages:
Primary
Secondary
http://www.ttuhsc.edu/som/obgyn/faculty/Prien...0Reproductive%20Physiology%20Lab%20Manual.htm (5 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Tertiary
Graafian: mature follicle that is ready to
ovulate
Atretic: follicles that degenerate and do
not progress to ovulation
After ovulation the ruptured follicle
becomes:
Corpus hemorrhagicum
Corpus luteum: produces
progesterone
Corpus albicans
Proestrus:
Estrus:
Metestrus:
Diestrus:
Anestrus:
Pregnancy:
Associated Structures: These are support structures for the reproductive tract.
Pelvic Bones or Girdle
Broad Ligament
Supports the uterus, oviducts, and ovaries
http://www.ttuhsc.edu/som/obgyn/faculty/Prien...0Reproductive%20Physiology%20Lab%20Manual.htm (6 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Contains the blood and lymphatic vessels and nerves
Uterine Artery
Ovarian Artery
Uterine Vein
Ovarian Vein
Utero-ovarian vein
Blood from this vein diffuses into the ovarian artery to
allow a direct path for
PGF2alpha (PGF2alpha has to go systemic
in the mare due to the lack of contact
between the utero-ovarian vein and the
ovarian artery)
Autonomic nerves
Sensory nerves
Arc-reflex
http://www.ttuhsc.edu/som/obgyn/faculty/Prien...0Reproductive%20Physiology%20Lab%20Manual.htm (7 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
http://www.ttuhsc.edu/som/obgyn/faculty/Prien...0Reproductive%20Physiology%20Lab%20Manual.htm (8 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Male Anatomy
Introduction: The structures of the male anatomy will be covered from the scrotum
and testes throughout the tract to the penis. The temperature control systems and
the sperm pathway will also be covered.
http://www.ttuhsc.edu/som/obgyn/faculty/Prien...0Reproductive%20Physiology%20Lab%20Manual.htm (9 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Scrotum: the sac-like structure that supports, protects, and regulates the temperature
of the testes
Made of two lobes
Homologous to the Labia majora in the female
The skin portion has a large number of sweat and
sebaceous glands
Tunica dartos is the smooth muscle portion
Testes: paired structures that are the primary sex organ of the male
Produces the male gametes (spermatozoa)
Produces hormones
Housed within the scrotum
Tunica vaginalis
An extension of the peritoneum that covers the testes
Tunica albuginea testis
The outer connective tissue layer
Parenchyma
The functional layer of the testes
Contains the Seminiferous tubules
Site of spermatogenesis
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (10 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Makes up 80% of testis weight
Germ cells (spermatogonia)
Become sperm
Seritoi cells (nurse cells)
Produces androgen binding
protein
Produces inhibin
Form the blood-testis barrier
Leydig cells (interstitial cells)
Located between the
seminiferous tubules
Produce testosterone
Cryptorchid
Condition where one or both testicles do
not descend into the scrotum
Unilateral cryptorchids are
fertile
Bilateral cryptorchids are
infertile
This condition can be inherited
Epididymis: paired convoluted tubules that transport sperm from the testes to the
ductus deferens
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (11 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Concentrate sperm
Storage of sperm
Maturation of sperm
Divided into three parts:
Caput epididymis (head)
Corpus epididymis (body)
Cauda epididymis (tail)
Spermatic Cord: paired structures that connect the testes to the urethra
Carries the nerve and blood supply to the testes
Testicular artery
Venous plexus
Lymphatic system
Nerves
Carry sperm to the urethra
Ductus derferens (also known as the vas deferens)
Support the testes
Temperature regulation
Controlled by the internal and external cremaster
muscles
Ductus Deferens: paired structures that transport sperm during ejaculation and
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (12 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
connect the epididymis to the pelvic urethra
Surrounded by a thick layer of smooth muscle
The ampulla is the enlarged end near the urethra
Absent in the boar
Urethra: single duct that transports both urine and sperm
Divided into two parts:
Pelvic urethra
Penile urethra
Accessory Sex Glands: glands located along the pelvic urethra
Contribute to fluid volume of semen
Provide nutrients, buffers, and other substances necessary for sperm
motility and
fertility
Secrete seminal plasma
Vesicular Glands (Seminal Vessicles)
Paired, lobular gland
Look like "cluster of grapes"
Contribute to over one-half of the total semen volume
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (13 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Secretes fructose and sorbitol (energy for sperm)
Secretes phosphate and carbonate buffers (protect
against shifts in pH)
Secretes the gelatinous material in stallion semen
Prostate Gland
Single gland
Secretes Na, Cl, Ca, Mg
Secretions cleans the urethra in the stallion
Bulbourethral Glands (Cowper’s Gland)
Paired glands
Secretions cleans the urethra in the bulls
Secretes the gelatinous material in boar semen
Penis: the organ of copulation in the male
Used by both the reproductive and urinary systems
Contains the penile urethra
Sigmoid Flexure
An s-shaped bend in the penis of bulls, rams, and boars
that allows the penis
to be retracted completely into the body by the
retractor penis muscles
Absent in the stallion
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (14 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Glans Penis
The free end of the penis
The shape varies depending on species
Homologous to the clitoris in the female
Most species have a fibro-elastic penis with some
erectile tissue
The horse however has a great deal more
erectile tissue
Cavernous Tissues
The erectile tissue that
engorges with blood during
sexual excitement
causing the penis to extend
(erection)
Corpus spongiosum penis
Corpus cavernosum penis
Ischiocavernosus muscle
Prepuce: encloses the free end of the penis
Also called the sheath
Homologous to the labia minora in the female
Sperm Pathway Out of the Body:
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (15 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Seminiferous tubules
Tubuli recti
Rete testis
Efferent ducts
Caput epididymis
Corpus epididymis
Cauda epididymis
Ductus Deferens
Ampulla
Pelvic Urethra
Penile Urethra
Temperature Control of the Testes: the testes need to be maintained 4 to 10 0F
lower than normal body temperature (91-97 0F)
Scrotum and Spermatic Cord
Tunica dartos muscle controls the scrotum’s surface
area
External and internal creamaster muscles control
proximity to the body
Cold weather causes these muscles to
contract decreasing the surface area
and drawing the testes closer to the body
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (16 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Hot weather cause these muscles to relax
increasing the surface area and
allowing the testes to swing down away
from the body
Sebaceous and Sweat Glands
In cold weather these glands are not as active
Hot weather causes these glands to become more
active allowing evaporation
to cool the scrotum
Pampiniform Plexus
The arteries that supply blood to the testes are very
convoluted and pass
through a pampiniform plexus of testicular
veins. This allows the arterial blood
entering the testes to be cooled by the
venous blood leaving the testes. As the
spermatic cord lengthens during hot
weather, more surface area is provided for
the heat exchange.
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (17 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Semen Analysis
Introduction: Semen analysis is a very important part of livestock production. Semen
is evaluated for artificial insemination, breeding soundness exams,
pre-purchase exams, and cryopreservation. Several factors are used
to evaluate semen. Each of these factors will be covered in detail.
Collection of semen for evaluation is usually done with the use of an
artificial vagina in stallions, rams, and bulls and by digital
manipulation in boars. It is also possible to use electroejaculation in
bulls and rams.
Gross Examination: this is the first step in semen evaluation and requires no special
equipment
Volume
Usually determined in milliliters
Gel-free volume is sometimes used
This is the volume after the gel-fraction is
removed
Volumes differ greatly between species
Low volumes can indicate a problem
Color
Milky
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (18 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Indicates high sperm concentration
Clear (watery)
Indicates low sperm concentration
Pink tinge
May indicate blood contamination
Yellow tinge
May indicate preputial fluid or urine
contamination
Activity (when observed closely)
Swirls of movement
Indicates high motility
No swirls
Indicates low motility
Consistency
Bull and ram semen will appear dense
Stallion and boar semen will be less dense
Debris
Dirt or other debris indicates that the male was not
properly cleaned or
careless collection
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (19 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Number of Live Sperm: the percentage of live sperm will be high than the percent
motile sperm
Eosin is a differential stain used to distinguish live form dead sperm.
Live sperm will appear clear
Dead sperm will be blue or green depending on the
background stain used
with the eosin
Process
1st a drop of semen is placed on a slide
2nd a drop of eosin is mixed with the semen and the
sample is then spread
across the slide
3rd slide is dried quickly usually on a hot plate
If dried slowly some sperm will die and be
stained
4th examine the slide under a microscope with 400x
power
5th count ten sperm from ten microscopic fields (total of
100 cells)
6th Figure % live
The number of live sperm divided by 100, multiplied by
100, gives the
percentage of live sperm (# live
sperm/100 x 100 = % live)
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (20 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Motility: the number of sperm that are moving
Circular and reverse movements are due to tail abnormalities
Lower than 40% motility is usually not considered suitable
Process
1st a drop of semen is placed on a slide
2nd add a coverslip
3rd examine slide under a microscope with 400x power
4th count ten sperm from ten microscope fields (total of
100 cells)
5th Figure motility
The number of moving sperm divided by
100, multiplied by 100 gives the
percentage of motile sperm (# moving
sperm/100 x 100 = % motility)
Forward Progression: a scale used to gauge the forward motion of the motile
sperm
Based on a five point scale:
5+ indicates moving extremely rapidly in a
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (21 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
forward motion
4+ indicated moving rapidly
in a forward direction
3+ moving forward with a purpose
2+ moving with no apparent direction
1+ indicates only a head or tail motion or moving in place
Morphology: the number of normal sperm
Process
1st heat kill the sperm on the slide used for motility by
placing it on a hot plate
for one minute
2nd count ten sperm from ten microscope fields (total of
100 cells)
3rd Evaluate the appearance
4th Figure % normal
The number of normal sperm divided by 100, multiplied
by 100, gives the
percentage of normal sperm (# normal
sperm/100 x 100 = % normal)
Concentration: the number of sperm per milliliter of semen
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (22 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
A MicroCell is a special slide used to count sperm. It has a measured
volume
(2ul) that allows for accurate counting of sperm.
Process
1st place a drop of semen into the chamber on one side of the slide
The MicroCell has two chambers allowing
for two semen counts per slide
2nd place the slide under the microscope and view at
100x power
3rd using the grid on the eyepiece, count the number of sperm in 10
squares
4th figure concentration per milliliter
If it is a non-extended semen sample the concentration
will be (# counted
x 106 = # of sperm/ml)
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (23 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
If the semen is extended at a 1:1 ratio
then the concentration will be (#
counted x 2 x 106 = # of sperm/ml)
If the semen is extended at a 2:1 (two
parts extender to one part semen)
then the concentration will be (# counted
x 3 x 106 = # of sperm/ml)
Densimeter
Machine that measures the density of a semen sample
and calculates the
concentration.
Spectrophotometer
Also known as a photoelectric colorimeter
Used by commercial semen production operations and
takes about one minute
Measures the light that passes through a semen sample
Calibrated with a sample of known concentration
Electronic Particle Counter
Most accurate way of determining sperm concentration
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (24 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Diluted sample of semen is passed through a capillary
tube so only one sperm
at a time can pass between two electrodes which
counts the sperm
Very costly
Viability: percentage of live, normal sperm
Figured by the multiplying the percentage of live sperm (as a
decimal number) by
the percentage of normal sperm (as a decimal number),
multiplied by 100, gives the % viability (% live x %
normal x 100 = % viability)
Acrosome Reaction: determines whether the acrosome cap has dissolved
The acrosome reaction must take place in order for the sperm to be able to go
through capcitation (a process necessary for fertilization).
Process
1st place a drop of semen that has been fixed in formalin
(formaldehyde) on a
slide
2nd add two drops of Chlortetracycline to the drop of semen and mix well
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (25 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Note: Chlortetracycline is a very potent neuro-toxin and it is
imperative that two sets of gloves are worn while handling
while
performing this procedure
3rd add a coverslip and a drop of microscope immersion oil
4th view with the oil immersion lens (1000x power) under a
florescent
microscope
All sperm will appear neon yellow
Non-acrosome reacted sperm will have an intact acrosome cap (sperm will
appear to have a distinct horizontal band across the middle of the head
Acrosome reacted sperm will have a head that appears
"faded" with no
distinct horizontal band
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (26 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (27 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Genetics
Introduction: Genetics are a critical part of reproduction. Each offspring is a product
of their parents. The DNA contained in both the sperm and egg is
responsible for information need to create an entirely new individual
of a species. Some basic genetics, chromosome numbers for the
different species, and some major genetically linked diseases will be
covered.
Basic Genetics
Gene: a sequence of DNA that codes for something (a trait), the basic unit of
inheritance
Locus: location of a gene on a chromosome
Allele: a form of a gene, two allele make up a gene (one on each homologous
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (28 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
chromosome)
Dominant Allele: an allele that will be expressed even if only one copy is
present, always represented as a capital letter
Recessive Allele: an allele that will only be expressed if two copies are present,
always represented as a lowercase letter
Co-dominant: both alleles are expressed (ex: roan)
Incomplete Dominance: one allele dilutes the other allele
Homozygous: having two copies of the same allele
Heterozygous: having one copy of each allele
Genotype: an animals underlying genetic make up
Phenotype: the characteristic that is expressed (due to the genotype)
Diploid: cells that have a 2n chromosome number (two sets of chromosomes)
n = the number of chromosome pairs in a species
Mitosis: cell division in which the diploid chromosome number is maintained
Haploid: cells (gametes) that have a 1n chromosome number (one set of
chromosomes)
Meiosis: cell division in the germ cells where the 2n chromosome number is reduced
to the 1n chromosome number found in the gametes
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (29 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Meiosis in the male:
Each primary spermatocyte under going meiosis results in four
spermatozoa
Meiosis in the female:
Each primary oocyte results in only one functional oocyte and three
polar
bodies
During fertilization (joining of the sperm and egg) the diploid (2n chromosome
number) is restored. Half of the genetic information (1n) comes from
the egg and the other half (1n) comes from the sperm.
Chromosome Numbers:
Human: 46 (23 pairs) Goat: 60
Horses: 64 Swine: 38
Ass: 62 Dog: 78
Cattle: 60 Cat: 38
Bison 60 Rabbit: 44
Sheep: 54 Chicken: 36
Genetic Problems:
Poly-spermia: More than one sperm fertilized an egg. Results in three or more sets
of chromosomes.
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (30 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Extra Chromosomes: Results from the incomplete separation of chromosome
pairs
during meiosis causing an additional chromosome in the egg or
sperm at conception.
Deleted Chromosomes: Results from the incomplete separation of chromosome
pairs during meiosis causing the sperm or egg at conception to have
too few chromosomes
Mutated Genes: can be due to carcinogens, radiation, toxins
Genetically Recessive Diseases: diseases that are only expressed if two copies of the
recessive allele are present. If only one copy is present, then the animal is a carrier for the
disease.
Dwarfism:
Palate- Pastern Syndrome:
Double Muscling:
Brain Hernia:
Hydrocephalus:
Spider Lamb Syndrome:
Leglessness:
Hairlessness:
Albinism:
Cystic Fibrosis:
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (31 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Sickle Cell Anemia:
Tay-Sachs Disease:
Embryo/Fetal Development
Introduction: It is important to have an understanding of the stages of pregnancy and
the development of the embryo/fetus and some of the problems that
can occur in order to be able to better manage the female and
offspring. A variety of species, including the human, will be used to
examine the development of the embryo/fetus.
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (32 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Terminology:
Zygote: As soon as the diploid number of chromosomes is returned
at
fertilization, what was once and ovum and sperm are now a zygote.
Embryo: From the two cell stage until differentiation is complete
the conceptus
is known as an embryo. This is usually not pat day 40
in the horse or day 60 in humans.
Morula: When the embryo has undergone three or four cell
divisions and is at
the 16 to 32 cell stage it is known as a morula. The
individual cells of the morula are known as blastomeres.
Blastocyst: After fluid begins to collect in the innercellular spaces
and forms a
cavity (called a blastocele) the embryo is known as a blastocyst.
Embryoblast: The cells of the inner cell mass which are located at
one pole are
now called the embryoblast.
Trophoblast: The outer cell mass that becomes the epithelial wall
of the
blastocyst is called the trophoblast. The trophoblast will
eventually become part of the placenta.
During the blastocyst stage the embryo hatches out of the Zona
Pellucida
Fetus: After differentiation has been complete and until parturition
the conceptus
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (33 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
is known as a fetus.
Development: Regardless of which species is being discussed, the gestation can be
broken down into three trimesters. The length of each trimester will vary
depending on the total length of gestation.
First Trimester: This is a very critical stage, because the all of the major organ
systems are developing and for the first few weeks, the female may not realize
that she is pregnant.
Begins at conception
Implantation of embryo
Day 6 to 7 in humans
Period of Organogenisis – development of all the major organ systems
Nervous system is 1st to begin forming and last to finish
Circulatory system is first to complete
Heartbeat detected at:
Day 16 in sow
Day 20 in ewe
Day 22 in cow
Day 23 in horse
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (34 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Day 25 in human
Digestive System
Reproductive System
Sexual Differentiation
Day 30 in swine
Day 35 in sheep
Day 45 in cow and horse
Maternal recognition of pregnancy
Period of greatest increase in cell numbers
Problems that can occur:
Lack of fertilization
Absorption of abnormal embryos due to problems such as poly-spermia and
genetic abnormalities
Implantation in the wrong place
Failure to implant
This is the period when the embryo is most susceptible to problems with
development
Most congenital malformations have their origin during this period
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (35 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Second Trimester:
Period of greatest size increase
Organ systems are growing and maturing
Third Trimester:
Period of greatest weight increase
Fetus is putting on subcutaneous fat layer
Fetus is preparing for birth
Organ systems are being completed
Lungs are preparing for breathing air
Nervous system will not be completed until after birth
Problems that can occur:
Premature delivery
Humans born during the sixth or seventh month have the most trouble
surviving
Animals born more than ~ two weeks early will have little chance of survival
Prolonged Pregnancy
Parturition
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (36 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Introduction: Many aspects of reproduction have been discussed up to this point,
including anatomy, semen analysis, genetics, and embryo/fetal
development. Now the end results of what we are trying to
accomplish with reproduction, getting an offspring on the ground,
will be discussed. In order to put an offspring on the ground,
parturition must be covered. Using the horse and cow as models,
the stages of parturition will be covered from the onset of labor to
delivery of the placenta.
Signs Labor is Imminent:
A few weeks (2 – 6 weeks) before giving birth, the udder will
become distended
7 – 10 days before foaling, the muscles around the tail head (croup)
relax and
shrink due to the general relaxation of the muscles and ligaments in
the pelvis
Mares may also leave the band prior to foaling, they generally want to be alone
when giving birth (most species are this way)
4 – 6 days before foaling, the mare’s teats fill
2 – 4 days before foaling, a waxy substance develops on the end of
the teats
At one day or less, the wax on the teats may fall off and milk can
start to drip
Stage One – Preparation: This is the first stages of parturition. Remember that
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (37 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
some animals that have had several offspring may not show any real
signs. They may just lay down and give birth.
Begins with uterine contractions and cervical dilation
Signs
Restless nervous actions showing discomfort
Cramps (signs of abdominal pain)
Some animals will kick at their bellies
Breaking out in a sweat
Walking round and round
Repeated standing and lying
This stage can last for several hours or only a few minutes, but
usually averages ~
20 minutes in the mares.
In cows this stage usually last for 2 – 6 hours during normal labor,
but can last
only 30 minutes or up to 24 hours
This stage end with the breaking of the allanto-chorion membrane
and the
expulsion of the water
This can be seen as a straw-colored fluid flowing from
the vagina
In some cases the "water bag" may actually protrude
from the vagina and the
fluid will be a dark amber color
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (38 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Stage Two - Delivery: The second stage is the delivery of the offspring.
This stage begins with the appearance of the amnion (whitish
membrane) or the
feet protruding through the vulva
This stage can last from 30 minutes to 4 hours in cows
Heifers may take longer
In the mare and sow expulsion should be faster because of the
danger of the
offspring suffocating due to the separation of the placenta
In the cow the placenta remains intact until the calf is
born
At this stage the female’s straining will be obvious as she tries to
expel the
offspring
The female may be lying or standing at this stage
This stage is complete when the offspring is delivered
It is important at this stage to be prepared to assist the animal in
cases of dystocia
or abnormal presentation
There are several abnormal presentations
Some of these can be corrected by
physical manipulation
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (39 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Others may need to have the offspring
pulled or delivered by C - section
Stage Three – Membrane Expulsion: This is the final stage of parturition.
The passing of the membrane (afterbirth) usually occurs within 30
minutes to 8
hours
Make sure to examine the placenta to make sure none it has been
retained
If the membranes are not expelled within 12 hours they are
considered to be
retained
Never try to pull a retained placenta out – no matter
how carefully this is
done, it can cause complications
Can give antibiotics, oxytocin, or prostaglandins to help
expel the afterbirth
When the membranes have been delivered, parturition is complete
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (40 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (41 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Bovine Reproduction
Introduction: Cattle are one of the most important of the domestic species in animal
science. Therefore it is vital that an animal scientist have an
understanding of the reproductive traits of this species so that the
best production decisions can be made. The normal vital signs,
reproductive traits, unique cow and bull traits, basic production
considerations, and important dates in early pregnancy will be
covered.
Vital Signs: In order in detect abnormalities or problems, it is necessary to know what
is normal.
Temperature 100.4 – 102.8 0F
Heart Rate 50 bpm
Respiration 30/min
Reproductive Traits:
Puberty
Dairy Cows 8 – 13 months of age
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (42 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
British Breeds 10 – 15 months of age
Zebu Breeds 17 – 27 months of age
Bulls 10 – 12 months of age
Gestation 278 – 293 days
Estrous 21 days
Estrus 12 – 18 hours
Ovulation 10 - 12 hours after end of estrus
Placentation: epithelio-chorial with cotyledons
Breeding Season: year long
Period of post-partum anestrus
Genetics 60 chromosomes (30 pairs)
Cow
Anatomy
Vagina
Site of semen deposition during natural mating
Cervix
Annular rings
Uterus
Bicornuate
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (43 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Uterine horns are fused giving the
impression of a large uterine body
Ovary
Almond-shaped
Ovulation occurs anywhere except the hillus
Signs of Estrus
Mount and stand for mounting by other cows
Restlessness and tail raising
Swollen vulva and clear mucus discharge
Vocalization (bawling)
Bull
Vessicular glands
Contribute to over ½ of the fluid volume of semen
Bulbourethral Glands
Secretions flush urine residue from urethra
Penis
Fibroelastic with a sigmoid flexure
Prepuce
Surrounded by long, tough preputial hairs
Ejaculation
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (44 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Last less than one seconds
Sperm
Cycle takes ~ 56 - 63 days
Output per day ~ 4 – 18 billion
Volume 3 - 10 ml
Concentration ~ 800 million – 1.2 billion per ml of
ejaculate
Motility 65 – 70%
Morphology 20% abnormal
Semen is collected with an Artificial Vagina or electroejaculation
Male to female ratio for natural service
Mature bull 40 – 50 per season
Production Considerations:
Breeding:
It is important to limit bull exposure to no longer than
90 days. This keeps the
calving season manageable, allows for
labor efforts to be concentrated, and
produces a more uniform calf crop.
Calving season can be scheduled to occur in either the
fall or the spring.
Estrus synchronization helps use labor more efficiently
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (45 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
when using AI.
Equine Reproduction
Introduction: The horse is a very important animal in the animal science industry.
Therefore, it is important to have an understanding of the
reproductive considerations in order to make the best production
decisions. The horse is the only domestic species that can
conceive while lactating. The normal vital signs, reproductive
traits, unique mare and stallion traits, and basic production
considerations will be discussed.
Vital Signs: In order in detect abnormalities or problems, it is necessary to know what
is normal.
Temperature 100.5 – 101 0F
Heart Rate 35 - 45 bpm
Respiration 12 – 36/min
Reproductive Traits:
Puberty
Mare 15 – 24 months of age
Stallion 13 – 18 months of age
Gestation 335 – 340 days (11 months)
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (46 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Estrous 21 – 23 days
Estrus 5 – 7 days
Ovulation 24 – 48 hours before end of estrus
Placentation: diffuse epitheliochorial with micro-cotyledons
Breeding Season: February to November (peak fertility May –July)
Long day seasonal breeder
Seasonal polyestrus
Period of winter anestrus
Genetics
Horse 64 chromosomes (32 pairs)
Ass 62 chromosomes (31 pairs)
Prezwalski 66 chromosomes (33 pairs)
Mare:
Anatomy
Vulva
The mare can have poor conformation of the vulva (slanted or
shelved) that allows feces contamination.
To prevent this a Caslicks operation
(stitching part of the vulva closed) can be
performed. This showed be opened 30
days prior to foaling.
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (47 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Clitoris
Found ~ 5cm inside the vulva
Harbors bacteria
Cervix
Has longitudinal mucosal folds
~ 5 – 7.5cm long and protrudes into the vagina
Vaginal fornix is an indentation around vaginal side of the cervix
site of semen deposition during natural mating
Uterus
Bipartite uterus.
Bifurcation - the division between the uterine horns.
Ovary
The ovary in the mare is a kidney bean shaped structure with an
ovulation fossa. This is unique to the mare
and the fossa is the only site where
ovulation takes place. (Ovulation can occur
anywhere on the ovary in other species
except the hillus.) The mare also is
different from other species because the
cortex is located on the inner portion of
the ovary while the medulla is on the
outside.
Signs of Estrus
Estrus in the mare can be detected by teasing (exposing her
to the stallion). When in estrus she will squat, have
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (48 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
frequent urination, lift her tail, and wink her clitoris.
Foal Heat
Another unique characteristic in the mare is the foal heat. It is the
1st post-partum estrus cycle. It is a fertile cycle and the mare can
conceive.
Can breed on foal heat if no dystocia or fluid in the uterus
More opportunities to breed
Can keep up with foaling dates
Usually occurs 7 – 9 days post-partum with ovulation occurring ~ day 11
Time to 1st post-partum estrus and ovulation shortens the later in the season that
the mare foals
Stallion:
Vessicular glands:
Large, produce the gel fraction (last portion of ejaculate)
Prostate
Secretes pre-sperm
Penis
Musculo-cavernosus
Largely erectile tissue that engorges with blood during sexual excitement to
produce an erection
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (49 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Glans penis is bell-shaped and flares (bells) during ejaculation
Ejaculation
Signaled by the flagging of the tail and the pulsing of the jets can be felt during
collection with the AV
Last ~ 10 – 15 seconds
Sperm
Cycle takes ~ 54 days
Production peaks at 7 –12 years of age
Output per day ~ 5 billion sperm
Volume 50 –75 ml
Concentration 30 –80 million per ml of ejaculate
Motility 60 – 100%
Morphology 20 – 30% abnormal
Semen is collected with an Artificial Vagina
Male to female ratio for natural service
2 year old stallion 4 – 7 mares
3 year old stallion 3 to 4 a week (10 –30 mares)
mature stallion once a day for 5 – 6 days or twice a day 3 times a
week (40
mares)
Flehmen response: a curling of the upper lip in response to estrus detection
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (50 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Vomeronasal organ is responsible for this
Production Considerations:
Spring transition
Period of irregular estrous cycles in the spring when the
mare is coming out of
anestrus
Light Program
Used to move spring transition to earlier in the season
Simulates lengthening daylight
Begin around Thanksgiving (last week in November)
Need 16 hours of light and 8 hours of dark
Recommend keeping lights on until 11:00pm
Constant light will cause very irregular estrous cycling
and is not good for the
stallion
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (51 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (52 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Ovine/Caprine Reproduction
Introduction: Sheep and goats are important species in animal science throughout the
world. It is important to understand the reproductive considerations
so as a producer, the best decisions can be made. Because sheep
and goats are similar in many respects, they will be covered
together. However, there are some traits in which they vary slightly,
as will be shown. The normal vital signs, reproductive traits, unique
traits of the ewe, doe, ram, and buck, and basic production
considerations will be covered.
Vital Signs: In order in detect abnormalities or problems, it is necessary to know what
is normal.
Temperature
Sheep 100.9 – 103.8 0F
Goat 101.3 – 103.5 0F
Heart Rate
Sheep 75 bpm
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (53 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Goat 90 bpm
Respiration
Sheep 19/min
Goat 15/min
Reproductive Traits:
Puberty
Ewe 7 -10 months of age
Doe 5 – 7 months of age
Ram 4 – 6 months of age
Buck 3 – 5 months of age
Gestation
Sheep 147 – 148 days
Goats 149 days
Estrous
Ewe 17 days
Doe 21 days
Estrus
Ewe 24 – 36 hours
Doe 30 – 40 hours
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (54 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Ovulation
Ewe late in estrus
Doe few hours after end of estrus
Placentation
Epitheliochorial with cotyledons
Breeding Season
Peaks from September to January
Can extend from late June to early April
Most are short day seasonal breeders
Genetics
Sheep 54 chromosomes (27 pairs)
Goat 60 chromosomes (30 pairs)
Ewe and Doe:
Anatomy
Vagina
Site of semen deposition during natural
mating
Cervix
Has annular rings
Serves as a sperm reservoir and filters out
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (55 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
dead sperm
Uterus
Bicornuate
In the ewe the uterine horns
are fused giving the
appearance of a larger
uterine body.
Ovary
Almond-shaped
Can ovulate from any place except the
hillus
Signs of Estrus
Ewe
Only exhibits signs of estrus when a ram is
present
Rubs the neck and body of ram
Smells rams genitalia
Shakes her tail vigorously
Doe
Signs of estrus are not as strong without a
buck present
Actively seeks out buck
Wags tail, bleats, urinates frequently
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (56 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Occasionally will stand to be mounted by
another doe, and will have some
mucous discharge and swelling of the vulva
Ram and Buck:
Vessicular glands
Small (4cm)
Prostate
Imbedded in the urethral muscles
Penis
Fibroelastic with a sigmoid flexure
Ram
Has a urethral process known as a filiform appendage that becomes
erect and
whirls in a circular motion during ejaculation
Ejaculation
Lasts less than one second
Sperm
Spermatogenesis takes ~ 49 days
Output per day ~ 8 - 12 billion sperm
Volume:
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (57 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Ram .5 -2 ml
Buck .5 – 2.5 ml
Concentration 2 billion per ml of ejaculate
Motility 75%
Morphology 10% abnormal
Semen is collected by electroejaculation
Male to female ratio for natural service per season
Ram Lamb 15 – 30 ewes
Yearling to five year old 25 –50 ewes
Aged Ram (6 year old +) varies depending on physical
condition
Flehmen response: a curling of the upper lip in response to estrus
detection
Vomeronasal organ is responsible for this
Production Considerations:
Tagging
Ewes should have wool around the dock and vulva
should be shorn prior to
breeding and lambing
Lambing
Ewe can be placed in lambing pens (at least 4ft x 4ft)
prior to lambing
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (58 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
should have a heat lamp
Apply iodine to naval cord
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (59 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Porcine Reproduction
Introduction: The pig is an important domestic animal. Due to the short gestation
length the sow can average 2.5 litters per year. The normal vital
signs, reproductive traits, unique sow and boar traits, and basic
production considerations will be covered.
Vital Signs: In order in detect abnormalities or problems, it is necessary to know what
is normal.
Temperature 101.6 – 103.6 0F
Heart Rate 60 bpm
Respiration 16/min
Reproductive Traits:
Puberty
Sow (Gilt) 5 – 7 months of age
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (60 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Boar 4 – 8 months of age
Gestation 114 days
Estrous 20 days
Estrus 48 – 72 hours
Ovulation mid-estrus
Placentation: diffuse epitheliochorial
Breeding Season: year long
Genetics
38 chromosomes (19 pairs)
Sow:
Anatomy
Cervix
Site of semen deposition during natural mating
Funnel-shaped with ridges that form a corkscrew
Uterus
Bicornuate
Uterine horns are long and convoluted
allowing more remove for the
multiple fetuses
Ovary
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (61 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Look like a cluster of grapes due to the growth of a large number of
follicles
Signs of Estrus
Swollen, pinkish red vulva
Will stand still when pressure is applied to the back (loin area)
Will mount other sows
Restlessness
Boar:
Prostate
Part of the glandular tissue
Secretes pre-sperm
Bulbourethral Glands
Secrete the gel fraction
Penis
Fibroelastic
Glans penis is corkscrew-shaped
Ejaculation
Last ~ 5 – 10 minutes
Sperm
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (62 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Cycle takes ~ 49 days (spermatogenesis 35 days)
Volume 150 – 250 ml
Concentration 200 – 300 million per ml of ejaculate
Motility 60%
Morphology 40% abnormal
Semen is collected by digital manipulation (gloved hand technique)
Male to female ratio for natural service
Young boars can pen breed 8 – 10 sows in a four week
period
Young boars can be handmated to 5 – 6 sows per week
Mature boars can pen breed 10 –12 sows in a four
week period
Mature boars can be handmated to 10 – 12 sows per
week
Production Considerations:
Reproductive Performance
Determined by pigs per sow per year
Performance declines after ~ 4.5 years
Post-weaning estrus is the first fertile cycle
Boars should always be considered dangerous and handled with care!
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (63 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Major Reproductive
Diseases
Introduction: The diseases that are associated with reproduction are very serious for
livestock producers. They can cause serious economic
consequences, because any diseases that causes problems with
reproduction can cause losses in offspring. Open females cost the
producer money.
Brucellosis: Also known as Bang’s disease or contagious abortion. In humans it is
called Malta fever. This is a reportable disease!
Cause
Gram negative intracellular bacteria
Brucella abortus (cattle) (can be the cause of fistulous
withers and poll evil in
horses)
Brucella melitensis (goats)
Brucella suis (swine)
Brucella ovis (sheep) – responsible for ram epididymitis
Brucella canis (dogs)
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (64 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
All types are infectious in domestic animal and humans.
Transmission
Ingestion is the most common method
Through licking or smelling of newborn calves form
infected cows, aborted
fetuses, membranes, or uterine fluids
Contaminated feed or water (from uterine fluids)
Can be through skin wounds, conjunctiva of the eye, milk from
infected cows,
vectors, copulation
Infected pastures are a source of contamination for 30 days
(remains viable in soil
and manure for ~3 days)
Signs and Symptoms
Inflammation of the genital organs and fetal membranes
Abortion may occur 2 – 3 months after infection
Sterility
Localized lesions in various tissues
Retained placenta will be thick and leathery
In the bull there will be orchitis and arthritis
Diagnosis
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (65 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Card test – also called the buffered antigen test
A field test performed at the sale barn by a veterinarian
Test all cattle over 20 months of age
Tube or rapid plate agglutination test
Mercaptoethanol test
Rivanol test
Brucella Milk Ring test
Treatment
No treatment is allowed!
All reactive cattle are branded and sent to slaughter!
Prevention / Control
Vaccination with RB-51 vaccine
Must tattoo right ear with RV and the year
Ear tag # and records sent to USDA
Only females between the ages of 4 – 10 months are
vaccinated
Performed by accredited veterinarian
Test and slaughter
Eradication program started in 1934
Chlamydiosis: Also known as Enzootic Abortion of Ewes (EAE). A highly
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (66 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
contagious disease that can cause severe economic losses in the sheep industry.
Cause
Chlamydia psittaci
Transmission
Ingestion of organism by licking aborted fetuses, membranes, or
fluid, or by
eating feed that has been contaminated
Can cause abortion in humans – Exercise extreme caution
with pregnant
women associated with flocks that have Chlamydial
abortions!
Signs and Symptoms
Abortion early in pregnancy
Weak, unthrifty, inefficient lambs
Diagnosis
Isolation of organism from the aborted fetus or membranes
Treatment
Antibiotics for two to three weeks
Prevention / Control
Vaccination
Remove aborting ewes from flock until all discharge is gone
Clean area where aborted fetus was thoroughly
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (67 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Feed ewes in bunks or feeders
Never keep poor lambs or ones that were weak at birth as
replacement animals
Epididymitis: inflammation of the epididymus. This is a serious concern in rams, but
it can occur in other species.
Cause
Actinobacillus ovis
Histophillus ovis
Brucella ovis
Transmission
The first two types are normally present in the prepuce, however
sometimes these
become pathogenic
The brucella ovis is transmitted either by homosexual activity or by
the ewes
during the breeding season
Signs and Symptoms
Heat, pain and swelling in the testicles during acute infections
The rear legs may appear lame due to the extreme discomfort
Enlargement and fibrosis of the epididymis
If scarring develops then the ram is basically useless
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (68 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
It is possible to surgically remove scar tissue blockage, however it is
cost
prohibitive
Wasting of testicular tissue
Can progress into secondary infections in the accessory sex glands
Diagnosis
Palpation of the testicles and epididymis for fibrotic lesions
Semen examined for white blood cells and abnormal sperm
Semen culture
ELISA enzyme linked immunoabsorbent assay
Treatment
Antibiotics work about ~ ½ of the time
Prevention / Control
Purchase disease free rams
Test rams
All positive rams should be culled
Test done every 60 days until conformation of
elimination
Genital Campylobacteriosis:. A venereal disease also known as Vibriosis.
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (69 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Cause
Bacteria
Campylobacter fetus venerealis (cattle)
Campylobacter jejuni (sheep)
Transmission
Coitus (copulation)
AI with contaminated semen or AI equipment
Sheep version is by ingestion, not venereal
Signs and Symptoms
No signs in bull
Temporary infertility
Irregular estrous cycle
Abortions may occur
Embryo reabsorbed (cattle)
Abortion by 5th or 6th month (cattle)
Abortion during last 8 weeks of gestation (sheep)
Herd history of cows returning for service after 60 days or longer,
large number
of open cows, overworked bull are signs
Diagnosis
Herd history and response to vaccine
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (70 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Isolation of bacteria in aborted fetus, lung, stomach, or placenta,
prepuce, vaginal
mucus of open cows
Treatment
Most cows recover in a short time if not re-infected
Some require 2 – 12 months for recovery
Can use penicillin or streptomycin
Vaccination is curative and preventative
In sheep outbreak produces immunity
Treat with antibiotics in feed
Prevention / Control
Vaccination of bulls and heifers
Females are vaccinated 6 weeks before breeding
Bulls require 2 injections and annual booster
Antibiotics in semen will kill organism
AI is best method of control
Vaccination and boosters for sheep
IBR/BVD: Infectious Bovine Rhinotrachitisis / Bovine Viral Diarrhea These diseases
are not true reproductive diseases, however, they do cause
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (71 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
problems with reproduction, so we will cover the reproductive
problems associated with them.
Cause
IBR – herpes virus
BVD – small RNA virus
Transmission
IBR
Contaminated feed or water (from nasal secretions)
Infected aerosol
Direct contact
BVD
Aerosols
Contact
Signs and Symptoms
IBR
Infection of the genital mucosa (vulva, vagina, penis,
prepuce)
Causes pustules and ulcerations
Abortion
3 – 5 weeks after cow has had respiratory
symptoms
Usually fetuses are 5 month or older
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (72 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Calves that are carried to term usually die
within five days of birth
BVD
Abortion 10 days to several months after disease
symptoms (diarrhea, fever
form 103 to 108 ° , ect.)
Superficial or ulcerated erosion of the mucosa (vulva
included)
Diagnosis
IBR from BVD
Symptoms
Isolation of virus
Differentiate between them
Treatment
IBR and BVD
Antibiotic for secondary infection
Prevention / Control
IBR
Vaccination of open cows and heifers
Disease can occur in vaccinated animals
BVD
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (73 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Vaccination only in healthy animals
Leptospirosis: Also known as Lepto, Infectious Jaundice, and Redwater.
Cause
Spirochete bacteria
Leptospira pomona
Leptospira hardjo
Leptospira canicola
Leptospira icterohemorrhagiae
Leptospira grippotyphosa
Leptospira bravasaliva
Transmission
Through skin or mucous membranes that come in contact with
infected urine
Infected animals are life long carriers
Can be transmitted to humans
Signs and Symptoms
Calves
Hemolytic icterus and hemoglobinuria
Elevated temperature (104 –106 ° F)
Anorexia
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (74 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Dyspnea
Anemia
Some die
Cows
Thick, yellow, blood-tinged milk
Abortion ~ 6 – 12 weeks after infection
Also common about 7th month into
gestation
"Abortion Storms"
Clear-red or port-wine colored urine
Seen more after high rainfall
Diagnosis
Herd history (no vaccinations)
Serology (blood samples)
Urine culture
Treatment
Antibiotics such as streptomycin, chlortetracycline, or oxytetracyclin
if given
early
Help eliminate carrier state
Prevention / Control
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (75 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Vaccination (5way)
Given every 6 months for 18 months then annually
Control rodents
Keep cattle away from infected water
Keep cattle and swine separated
Trichomoniasis: Also known as Trick. A contagious venereal disease in cattle
Cause
A pear-shaped flagellated protozoan known as Tritrichomonas
Transmission
Normally only spread during coitus (copulation)
90% of the females bred by an infected bull become
infected
It is possible to be spread by AI, however, the antibiotics used in
semen extenders
reduce the chance of this
Signs and Symptoms
Infertility
Repeat breeding
Irregular estrous cycle
Fetus death ~ 50 to 100 days after conception
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (76 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Some cows will have
Pyometra
Vaginitis
Cervicitis
Endometritis
A herd history of reduced pregnancy rates, wide spread calving
season, wide
variation in weaning weights, reduced calf crop – are all signs of trick
Diagnosis
Herd history
Finding of organism in placental fluid, stomach contents of aborted
fetus, or
uterus after abortion
Finding the organism in the prepuce, sheath, or penis
Treatment
Cows will usually clear themselves of infection in 90 days if given
sexual rest
Best to AI for at least two years
Treatment of bulls is not usually done because of expense
Should be slaughtered
Prevention / Control
Vaccine is now available
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (77 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Give twice a year to cows
Not tested in bulls yet
Reproductive Technology
Introduction: Ever since man domesticated animals, he has been manipulating the
reproduction in these animals. It started by selecting which animals
were to become parents. It continued with selection, culling, and
specific breeding programs. Now, he is able to use modern
technology to alter the reproduction in these animals. These
advanced technologies used in reproduction will be covered.
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (78 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Artificial Insemination: A process of introducing semen into the female
reproductive tract by an artificial means (not with natural deposition by the male).
History:
1st reported use was in the 1300 in Arabian horses
1st documented case was in 1780 by an Italian physiologist, Spallanzani
Started with several amphibians
Then with dogs
U.S.S.R.’s Ministry of Agriculture trained veterinarians in AI
1st Artificial Vagina 1914 (used for dogs) @ the University of Rome
Later AV’s were developed for use in larger species
1940’s the electroejaculator was developed
Benefits:
More offspring from superior males
Many bulls can produce enough semen in one year to inseminate 40,000 females
Many boars produce enough semen in one year to inseminate 1, 500 to 2,000
sows
Helps control the spread of venereal disease
Elimination of dangerous males (with shipped or cryopreserved semen)
Disadvantages:
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (79 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Labor for collection and insemination
Estrus detection a must
Some special facilities need (chutes, stocks, equipment)
Training needed to be proficient at AI
Techniques:
1st technique involved putting the semen into the vagina
2nd technique used a speculum placed in the vagina, light used to see posterior
cervical opening, glass rod inserted into the cervix
where semen was deposited
1937 Danish veterinarian developed the recto-vaginal (cervical fixation)
technique to be used in cattle
One hand is inserted into the rectum to manipulate cervix while the
other hand
guides the AI rod (insemination tube) into/
through the cervix where the semen is
deposited (into the anterior cervix or the
uterus)
This technique is still used today!!
Mares: AI is performed by inserting a hand into the vagina where two or three
fingers are inserted into the cervix to help guide the rod. The semen
is then deposited into the uterus.
Ewes and Does: can use three methods
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (80 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Speculum used to see posterior cervical opening, semen is deposited into cervix
A ball-tipped, 17 gauge needle attached to the end of the
insemination catheter is
passed through the cervix and semen is deposited into the uterus
Transcervical: cervix is grasped with Bozman forceps, to stabilize it, then a fine
insemination rod is "threaded" through the cervix where
the semen is deposited into the uterus
Laparoscopy Intrauterine insemination: a local anesthesia is applied to the
site
(usually in the flank) where the cannula is to be
inserted. The reproductive tract is located with the
laparascope and semen is injected into the lumen of the
uterus.
Sows: AI in the sow is very simple due to their cervical anatomy
The AI rod has a cork screw shape (similar to the boar’s penis) that is screwed
into the cervix and the semen is deposited into the uterus
Estrus Synchronization: The artificial manipulation of the estrous cycle in order to
cause the female to come into estrus (heat) at a more convenient time for the
producer. It is usually performed on several females at the same time.
Advantages:
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (81 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
More efficient use of labor for artificial insemination
More efficient use of labor at calving
Can decrease number of non-productive days
Necessary for embryo transfer
Time for estrus detection reduced
Disadvantages:
Having a number of heifers to "break in" to the milking routine at the same time
can be a problem for dairy producers
Labor – working animals to place and remove implants, give injections, or place
in feed
If using PGF method – can cause abortions
Synchronization Methods:
Prostaglandin (PGF)
Brand Names
Lutalyse
Bovilene
Estrumate
Usually given as two injections 11 days apart
Causes regression of the corpus luteum
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (82 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
CL must be present for it to work
Breed on estrus after 2nd injection
Progestens
Brand Names
Synchro-Mate B
Melengestrol Acetate (MGA)
Regumate
Synthetic progesterone
Replaces corpus luteum
Estrus occurs after withdraw of product
Can be given orally or implanted
Can be used in conjunction with PGF
Superovulation: A procedure where the female is treated with hormones to cause the
production of several ova instead of just one
Advantages:
Increases the use of superior females
Disadvantages:
Must inseminate with two to three times more semen and inseminate more
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (83 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
frequently
Labor – must handle animal more
Must use estrous synchronization and embryo transfer which are already labor
intensive
Ewes
A subcutaneous injection of 1000 IU of eCG (equine chorionic
gonadotropin,
PMSG) is given on day 12 or 13 of the estrous cycle to cause
superovulation
Averages ~ 12 ova ovulated
Cows
Treatment with FSH
Care should be taken not to over-stimulate the ovaries. This can result in
permanent damage
Several protocols have been used
eCG (PMSG) at a dose of 2000 - 2500 IU
Only used for research or in countries where eCG is approved for use in cattle
Averages ~ 12 ova ovulated
Sows
Subcutaneous injection of 750 – 1500 IU of eCG (PMSG) on day 15 of estrous
cycle (dose depends on size of animal)
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (84 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
A subcutaneous injection of 500 IU of hCG is given at the onset of estrus to cause
ovulation
Mares
Reliable superovulation has not yet been achieved
Embryo Transfer: the transfer of embryos from a donor (usually a superovulated
female)to a recipient
Embryos should be transferred within two hours or cooled to prevent degeneration
Embryos can be kept at 15 – 25 C0
Embryos can be frozen and stored for later use (Cryopreserved)
Ewes
Recovery of embryos is usually done surgically
Can be flushed from the oviduct and collected at the infundibulum
Cows
Recovery can be done with a Foley catheter
Lets fluid in and collects it when it comes out
Done one horn at a time
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (85 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Sows
Recovery can be done surgically before the embryos reach the uterus
Mares
Same as the cow, but both horns are flushed at the same time
Transfer Methods
Trans-cervical
Embryos are transferred into recipients using an AI rod
Embryos are placed in the uterine horn adjacent to the CL
Cervical Bypass
Embryo are placed in the uterus by using a long hypodermic needle that is
inserted through the vaginal wall and into the uterus
(bypassing the cervix)
Flank Surgery
Embryos are placed into the uterus using laparoscopy through the flank
In Vitro Fertilization: a process whereby the oocyte is fertilized outside the body
1st reported birth of an offspring from IVF occurred in 1959 with a rabbit
IVF is now common in many species including humans
Techniques
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (86 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
ICSI – Intercytoplasmic Sperm Injection
The most common technique used today
Involves injection of one sperm into the oocyte
SZI – Sub-Zonal Injection
Involves the injection of five sperm under the zona pellucida into the
perivitalin space
PZD – Partial Zonal Dissection
This technique is no longer used due to the large number of multi-sperm
fertilization
Cryopreservation of Semen: a process of storing semen by freezing
Extender must be added so semen can be divided and protected
Extender must contain:
Buffer – to protect against shifts in pH
Cryoprotectants – substances such as glycerol or DMSO are used to protect
sperm during freezing and thawing
Semen must equilibrate for at least four hours (and up to 12 hours) to
allow the glycerol to penetrate the sperm
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (87 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
Nutrients – for sperm energy
Antibiotics – for protection against microbial contamination
Usually stored in liquid nitrogen
Semen is stored in straws or ampules
Embryo Cloning (Twinning): the creation of multiple sets of genetically identical
embryos from one original embryo
Can be used to create identical embryos for research to eliminate genetic variation
Can be used to duplicate transgenic animals
Procedures for Creating Embryo Clones
Embryo Splitting – also known as embryo bisecting
Involves the separation of early embryonic cells (usually at the 4 or 8 cell
stage, but no later than the 16 cell stage)
Problems with smaller embryos and limited numbers lead to the development
of nuclear transfer
Nuclear Transfer
This process involves removing the nucleus from an unfertilized oocyte and
replacing it with the nucleus from a donor
cell (from an embryonic cell at the 4, 8, or
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (88 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
16 cell stage) creating a new embryo.
Theoretically an unlimited number of copies can be made, however,
realistically ~1000 can be created from
one embryo with only ~ ½ of those
surviving
True Cloning: the production of a genetically identical copy of an adult organism
1962 – John Gurdon cloned frogs from the gut cell of an adult frog
Clones nerve grew past the tadpole stage
1996 – Dr. Ian Wilmut, Dr. Keith Cambell, and their associates at the Roslin Institute
created the first cloned mammal, Dolly
1998 – Dr. Ryuzo Yanagimachi at the University of Hawaii created three generations
of cloned mice
1998 – Dr. Yomiuri in Japan created twin female calf clones from adult somatic cells
1998 – a clone of the last surviving member of a cattle herd from Enderby Island was
created
There are several potential benefits that can come from true cloning if some of the
obstacles can be overcome
Embryo Sexing: the determination of which sex an embryo is could be very helpful
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (89 of 90) [6/10/2008 12:00:29 PM]
ANSC 3401 Reproductive Physiology Lab Manual
to producers that are performing embryo transfer by allowing them to choose
more females for the cow/calf operation
Separation of Male (Y) and Female (X) Producing Sperm: this would
allow producers to select male or female producing sperm for AI
Pregnancy Testing: there are several techniques that can be used for pregnancy
detection
Progesterone Assays
Rectal Palpation
Ultrasound
Detecting pregnancy is very important to the producer because the loss of money on
every open (non-pregnant) animals
http://www.ttuhsc.edu/som/obgyn/faculty/Prie...Reproductive%20Physiology%20Lab%20Manual.htm (90 of 90) [6/10/2008 12:00:29 PM]