4. Dimensions(μm) of Mammalian
Spermatozoa
species Head Mid piece Principle
piee
Total length
Length width Length Width
Bull 9 4.5 14 42 50-70
Boar 8.5 4.2 10 36.1 54.6
Ram 8.2 14 42 60-70
Horse 6.41 3 50
Rat 7.9 3.2 18.4 1.3 96 123
Dog 5.9 3.9 10.6 45.8 62
Cat 5.76 2.5 8.34 0.77 46.25 59
5. Principal part of spermatozoa
• Nucleus
• Acrosome
• Connecting piece
• Middle piece
• Principal piece
• End piece
HEAD
TAIL
6. Head
⚫It is the anterior most part of the spermatozoa.
⚫The head of the spermatozoais elongated and ovoidin
shape.
⚫The head length and width of spermatozoa is varied
species to species.
⚫The head is formed of acrosome and nucleus.
7. Nucleus
⚫The head nucleus is entirely filled with nearly homogenous
nuclear material.
⚫Haploid chromosome are present in the nucleus.
⚫DNAsurrounded bythe nuclear membranes.
8. Acrosome
⚫ The anterior 60 percent of the nucleus covered byacrosome cap or head
cap.
⚫ The acrosome is formed by golgi body and it is bounded byunit
membrane.
⚫ Outer membrane of acrosome cap is identical with galea capitis.
⚫ Acrosome cap thickness is 0.1 microns.
⚫ Followingenzymes are present in the acrosome –
Acid hydrolases,Acid phosphatase, Hyaluronidase,Acrosin
etc.
9. ⚫The anterior part of the nuclear membrane or the inner
membrane of acrosome,is modified to form the
perforatorium.
⚫Cell membrane encloses the head,body and tail of the
spermatozoon.
⚫The membrane is closely applied to the cell at the
anterior portion ofthe acrosomecap, the postnuclear
cap and at Jensen’s ring.
10. ⚫Detachment occurs byrupture of cell membrane and outer
membrane of the acrosome cap in the equatorial zone.
⚫The detached outer membranes of the acrosome may
break into two halves,or remain joined and appear as
“bathing –cap”shaped structure.
12. Tail
⚫It is the longest part ofsperm.
⚫It contains axial filaments.
⚫Axialfilamentiscoveredbyasheath, whichismadeupof9
singlet fibers.
⚫The morphological point of view tail consist – Neck,middle
piece, principal piece and end piece.
13. Neck
⚫It is the smallest part of spermatozooa.
⚫ In neck of sperm 2 centrioles are present.
Proximal centriole
Distal centriole
⚫Proximal centriole lies in adepression in the posterior surface of
the nucleus.
⚫Proximal centriole is perpendicular to main axis of the sperm.
14. ⚫The proximal Centriole hasno active function in the
spermatozoon but is apotential activist within an egg during
first cleavage division of the fertilized egg
⚫Distal centriole is alonglongitudinal axis ofthe sperm.
⚫Distal centriole acts asbasal body and gives rise to axoneme
of the sperm-tail.
⚫Axial filament of sperm structure as 9 + 2 manner.
15. Middle piece
⚫ Middle piece is located between the neck and the
annulus.
⚫ It containsthe mitochondrial sheath, ahelix 80 to 100
mitochondria wrapped around the axoneme.
⚫ The axoneme of the mammaliansperm is surrounded bynine
outer dense fibers which are also called the coarse or accessary
fibres.
⚫ The mitochondrial sheath is believedto be the source of energy
(A
TP) for sperm motility
.
16. ⚫Middle piece of sperm have 9+9+2 fibrous pattern.
⚫At the junction of mid piece and principal piece is present
the annulus which isalso known as the ringcentriole or
Jensen’sring.
⚫This ringcentriole prevent the mitochondria toward the tail
reason.
17. Principal piece
⚫The Principal piece continues posteriorly from the annulus
and extend to the near the end of tail .
⚫The principal piece of mammalian spermatoza is surrounded
by afibrous sheath.
⚫ Fibrous sheath is composed of a series of circumferentially
oriented ribs that extend half way around the tail end in two
longitudinal columns.
18. End piece
⚫ The end piece or terminal portion of the tail is about 3 to 4 microns in length
and consist of the terminal portion of the fibrils coveredby the cell membrane
but fibrillar coil sheath of the tail is absent.
19. Axenome filament
⚫Runs throughout the tail.
⚫The core ofthe axenome consists oftwo central microtubules
surrounded byarow of nine doublet microtubules.
⚫one microtubule of each doublet is complete, having 13
protofilaments, the other is C-shaped and has only 11
protofilaments .
⚫which are made exclusively of the dimeric protein tubulin.
20. ⚫Dynein arm motor complexes allow microtubule to slide
against each other.This causes thesynchronised shortening
and extension of the microtubule on opposite site of the
axoneme allowing the flagellar to bend.
⚫Alteration of this bending action creates the beating motion
of aswimming sperm.
⚫Radial spoke regulate the motion ofaxoneme.
22. Points to remember
⚫The head and tail are principle part of spermatozoa.
⚫Acrosomal cap and nucleus are part of head.
⚫Tail are subdivided in to the four part Neck, middle piece,
principle piece and end piece.
⚫Outer coarse fiber are present on the surface of mid piece
and principle piece only
.
⚫Mid piece and principle piece have9+9+2 fibrous
arrangement.
⚫End piece have 9+2 fibrous arrangement.
⚫Tubulin and dynein are the axonemal protein.
29. Pre-copulatory Stage
• Courtship
Species specific events
Sniffing of the vulva by male
Urination by the female
Flemen lip curl (exept BOAR)
Chin resting on female rump
Increased phonation
Male checks for female lordosis
30. Copulatory Behavior (cont.)
• Varies among species
• Short copulaters (1 - 3 seconds)
Bull
Ram
• Sustained copulater (5 - 20 minutes)
Boar
• Intermediate copulater(20 to 60 seconds)
Stallion
31. Pre-copulatory Stage
• Search for sexual partner
In female animals - limited to estrus
In primates can occur at any time
In male can occur at any time
Increased physical activity
• Involves all of the senses
1. Visual (Sight)
2. Olfactory (Smell)
3. Phonation
4. Tactile
32. Precopulatory Stage (cont.)
• Sexual arousal
• Female
• Lordosis
• Presents hindquarters to male
• Male
• Erection
• Penile protrusion
33. Copulatory Behaviour
• Erection and protrusion of penis (exeptional)
• Mounting and fixation
• Intromission
• Ejaculatory thrust and ejaculation
34. Erection and protrusion of penis (exceptional)
• In most animals, Erection and protrusion of penis from prepuce, occurs
before mounting
• Exception - camel the full protrusion occur after mounting
35. Mounting and fixation
• Mounting – Male fixes his fore legs around the hind quarter of female
grasp her firmly which follows pelvic thrusts
• In she camel the mounting occur in sitting position
• In cat and Rabbit the male bite the scruff (behind the head or at the neck)
36. Intromission
• Contraction of the abdominal muscle (rectus abdomenus) results in direct
contact (apposition) of the pelvic region of male to the female external
genitalia followed by penile insertion
Ejaculatory thrust and ejaculation
Involuntary, pulsatile release of semen in female reproductive tract
37. Post copulatory behaviour
• Dismounting
• Memory
• Refractory Period/ Refractoriness
• Dismounting – Retraction of penis into the
prepuce
38.
39. Refractory Period/Refractoriness
• Most animals show sexual inactivity immediately after
copulation
• Ram and buck in exception usually lick the penis after
ejaculation.
• Cats smell the site of copulation.
• The duration of this period is variable and modified by
environmental stimuli (5 min to one hour )
• This period is decreased by changing the female
40. Pre-Copulatory behavior in bull
• Courtship
• Grazing with cow
• Guarding the cow
• Liking cow vulva
• Dribbling prostate secretion
• Flehman’s reaction
49. Tomsexualbehavior
• Olfactory cue in female
urine
• Calling the female
• Sniffing
• Holding the queen
neck
• 10 seconds
• Lick its penis
50. Semen evaluation
• Provides information on the complex of sexual function
• The semen is the normal discharge of the male at time of mating.
• It is a suspension of spermatozoa in a fluid medium called seminal
plasma.
• Spermatozoa originates from the testes, primary sex glands and stored in
the epididymis, constitute 10% of the total volume
• Seminal plasma is a mixture of secretions from seminal vesicles, cowper’s
gland, prostate gland, ampullae and epididymis.
• Secretions of seminal vesicles form nearly 55% of the total volume of the
bull semen
• It takes about 60 days before spermatozoa appear in the ejaculated
semen.
51.
52. Care during semen handling
• Hygiene of AV - clean and free from contaminants excessive petroleum
jelly powder present on new liners and antiseptics and chemicals of any
kind.
• At the time of collection excessive dirt and debris should be kept
• Immediately after collection the semen vial should be placed in a water
bath at 37°C.
• Overheating and rapid chilling of semen should be avoided as it affects the
semen quality.
• Too much agitation and shaking of semen should be avoided.
• Exposure of semen to sunlight should be avoided ??????
53. Battery of tests
• Macroscopic examination
• Microscopic examination
• Biochemical tests
• Resistance to environment
• Physical evaluation
• Chemical Evaluation
• Biological Test
54. Macroscopic examination
1. Volume
• Remains fairly constant for each species.
• Varies among individuals and between ejaculates within the same
individual.
• Volume increases
Increase in age and body size of animal
General reproductive health
Vigour and frequency of service.
Teasing and stewing of bulls are practiced to increase the volume of
semen.
Semen volume increases up to 6-8 years of age.
55. Macroscopic examination
• Decreased in semen volume
• Young or old males
• Reproductive / general illness
• Males used excessively
• Incomplete ejaculation or failure of ejaculation
• Bilateral seminal vesiculitis
56. Macroscopic examination
Species Volume of Semen
Average (ml) Range (ml)
Bull 4 1-15
Stallion 70 30-250
Ram and Buck 1 0.7-3
Boar 250 1.25-400
Dog 10 1.25
Cat 0.04 0.01-0.12
Fowl 0.75 0.25-2.0
Elephants - 50-100
Volume of semen in difference species
57. Macroscopic examination
2. Colour
• Bull and buck semen - Milky white, creamy or opaque.
• Buffalo semen is whitish when compared to bull semen.
• In stallion, boar and dog - Pearly white to grey and translucent.
• Yellow colour of semen is normal in some bulls (spl. indegenous)
• Due to riboflavin content,secreted from ampulla or seminal vesicles
• The highly concentrated semen will be creamy in colour and if there is
only very few or no sperms then the colour will be watery.
58. Macroscopic examination
2. Abnormal Colour
S.no Abnormal Colour Etiology
1 Brownish Orchitis (Blood pigments)
2 Dark red to pink blood Hemorrhage in male reproductive tract
3 Yellow green Pseudomonas aerogenosa infection - pus
This colour appears on keeping semen some time after
collection
4 Light brown Contamination with dung
5 Dull and dirty white Increased number of spermatogenic cells
6 Yellow Presence of urine
7 Chunk clots/Curdy
appearance
Infection (Vesiculitis/Brucellosis)
59. Macroscopic examination
3. VISCOSITY AND DENSITY
Viscosity is assessed by using semen delivery pipettes. Viscosity increases
with sperm concentration
The specific gravity of bull semen is 1.0361 There is a positive correlation
between specific gravity and sperm cell concentration.
Colour Density Grade
Creamy DDDD
Milky DDD
Thin milky DD
Translucent & cloudy D
Watery O
60. Macroscopic examination
• Certain pathological conditions of testis and accessory sex glands may
affect the consistency of semen
Pathological condition Consistency of semen
Epididymitis Less milky semen
Catarrhal conditions of accessory sex glands Thick viscous semen
Seminal vesiculitis Purulent flocculi
61. MICROSCOPIC SEMEN EVALUATION
• Mass Motility
• The collective movement of sperms or their wave motion is called as mass
activity
• It is estimated by keeping a semen drop on a warm glass slide and
examining under low power microscope with out putting coverslip.
• Mass activity may be classified into five grade scale depending upon the
wave motion and rate of sperm activity.
• 5 and 4th grade samples are acceptable. Others should be discarded.
62. MICROSCOPIC SEMEN EVALUATION
• Mass motility
S.No. Findings Description Grade
1 Very vigorous forward motion, extremely rapid waves
and eddies, about 90-100 % active sperms.
Excellent 5
2 Vigorous, progressive movement with rapid and
abruptly forming waves and eddies, about 70-80%
sperms are motile.
Good 4
3 Progressive rapid movement of sperm, slow moving
waves and eddies, 50-60% sperms are motile.
Fair 3
4 Oscillatory or rotary movement, no waves and
eddies, 30-40% progressively motile sperms.
Poor 2
5 Stationary bunting or weak rotary movements, 10-
20% scattered progressive sperms.
Very poor 1
6 Immotile sperms. All dead 0
63. MICROSCOPIC SEMEN EVALUATION
2. INDIVIDUAL MOTILITY
• Motility is the most common and extensively used tool for estimating the
semen quality.
• The movement of individual sperm - the individual motility.
• To assess the individual motility diluted semen (diluted with physiological
saline or 3% sodium citrate or Tris buffer or Tris-egg yolk extender) is kept
on a warm slide and covered with cover slip.
• The slide is kept on the stage warmer of the phase contrast microscope
and examined under high power.
• Progressive movement - sperms with very rapid straight forward direction
64. MICROSCOPIC SEMEN EVALUATION
INDIVIDUAL MOTILITY
• Circular movement - sperms with movement in circular path – Cold shock
• Reverse movement - sperms moving in reverse manner
• Oscillatory movement - sperms with jerky movement – Energy difficiancy
• The progressively motile sperms are only taken into account, while
estimating the initial motility.
• The progressive sperms will cover a distance of 100-120 µ in a second.
65. MICROSCOPIC SEMEN EVALUATION
• Based on progressive motility, the semen samples are graded
as follows.
S.No. Progressive motility Grade
1 90-100% Excellent
2 70-80% Good
3 50-60% Fair
4 30-40% Poor
5 0-20% Very poor
A good semen sample should have an initial motility of 70%.
67. ESTIMATION OF SPERM CONCENTRATION
Haemocytometer
• It is a very old method used to assess the sperm concentration but
is most accurate method.
• The procedure is as same as RBC estimation.
Advantages
• Accurate method
• It is used when few bulls requires estimation
Disadvantages
• Requires skill
• The procedure is time consuming.
• So it cannot be used in places where large samples are processed.
68. CONCENTRATION ESTIMATION BY
HAEMOCYTOMETER
• Materials required
• Semen sample (fresh/frozen)
• Haemocytometer set
• Phase contrast microscope
• Watch glass
• Dilution fluid (0.1% formal saline or distilled water )
• Eosin powder
• Blotting paper
69. Concentration estimation by haemocytometer
Procedure
• Mix the semen sample gently to get uniform distribution of sperms.
• Place the semen in a sterile watch glass.
• Put a speck of eosin powder and mix it with semen.
• Aspirate the semen from watch glass into a RBC pipette upto 0.5 mark.
• Clean the tip of the pipette with blotting paper.
• Draw the dilution fluid in the same RBC pipette upto 101 mark.
• Roll the pipette between palm of the hands for 2 minutes to ensure
through mixing of the fluid and semen.
• Discard first few drops.
70. Concentration estimation by haemocytometer
Procedure
• Charge the haemocytometer by releasing the fluid below the coverslip
which is placed over the haemocytometer.
• While charging overflowing and air bubble formation should be avoided.
• Wait for 1-2 minutes for the sperms to settle.
• Examine the charged haemocytometer for under low power and then in
high power.
• The sperm counting is done in RBC chamber.
• Count the number of sperms in left top, right top, right bottom, left
bottom and center squares of RBC chamber and calculate the
concentration.
72. Concentration estimation by haemocytometer
Counting 5
chambers
Number of sperms in 1 ml of
semen
= N x 10000 x 1000
= N x 107 millions
73. Estimation of sperm concentration
• Cell volume method
• The semen is centrifuged immediately after collection.
• The sperms will deposit at bottom and the seminal plasma will come to
top.
• The packed volume of sperms is measured and based on this the
concentration is estimated.
• This is not an accurate method because the materials other than the
sperms will interfere with the result.
74. Estimation of sperm concentration
• Calorimeter
• Here the optical density of the semen sample is measured and
from which the concentration of the sample is arrived.
• The colorimeters are designed to measure the percentage of light
transmitted through a light absorbing media can be used.
• The percentage of transmission is a function of the concentration of
the light absorbing agents in the medium.
• This principle is applied to estimate the sperm concentration in a
semen sample.
75. Estimation of sperm concentration
• Calorimeter
• The percentage of transmission recorded has to be converted as sperm
concentration.
• Standardization with haemocytometer
• Based on this a working chart has to be prepared and it can be used
for routine concentration.
• The instrument has to be calibrated once in a month.
Diluted semen
Meter reading
76. Estimation of sperm concentration
Calorimeter
• Advantages
• Faster than other methods.
• Results are reliable
• Disadvantage
• Initial standardization requires time
• Operation requires time and skill.
• The media used for suspending the semen should dust free otherwise the
results will be wrong.
77. Estimation of sperm concentration
• Photometer
It is the advanced form of
colorimeter.
Principle - same as colorimeter.
Instrument itself will display the final concentration, dilution rate and
number of doses can be made.
It is faster than the colorimeter.
The instrument has to be calibrated once in two weeks with
haemocytometer to ensure better working condition.
78. Estimation of sperm concentration
• Opacity tubes
• Brown’s opacity tubes have been used for estimating sperm
concentration.
• Opacity increases with increase in concentration of semen.
• Computer Assisted Semen Analyzer (CASA)
• It is the most advanced method.
• Instrument is costly.
79. Normal sperm concentration in different
species
.No. Species Concentration
1 Cattle bull 1200 (800-1400 millions/ml)
2 Buffalo bull 800 (600-1200 millions/ml)
3 Stallion 250 (200-600 millions/ml)
4 Ram and buck 3000 (2000-4000 millions/ml)
5 Boar 250 (200-500 millions/ml)
6 Dog 250 (125-500 millions/ml)
80. Concentration related nomenclature
S.No. Terminology Explaination
1 Normozoospermia Normal sperm concentration
2 Oligozoospermia Reduced sperm concentraion
3 Polyzoospermia Increased sperm concentration
4 Azoospermia Zero sperm concentration
81. Viability of spermatozoa
• Traditional method – EOSIN NIGROSIN
• Principle
• The live and dead sperm is assessed by a vital stain - eosin-nigrosin which
was found by Hancock during 1951.
• The eosin is a vital stain which can only pass through the loosely integrated
plasma membrane of dead sperm and stains it as pink color.
• The nigrosin is a negative stain which gives background color.
• Initially - Mayer (1947) by using eosin (vital stain) and opal blue
(background stain).
• Later Hancock (1951) used eosin (vital stain) and nigrosin (background)
82. EOSIN NIGROSIN
• Materials required
• Semen sample (fresh/frozen)
• Glass slides
• Eosin stain (5%)
• Nigrosin stain (10%)
• Immersion oil
• Phase contrast microscope
• Preparation of 5% eosin stain
• Weigh 5 gm eosin powder, put in pestle and mortar.
• Prepare 2.9% sodium citrate solution, boil it.
• Add the boiling solution to stain and grind it well.
• Finally filter and store it at 4 ⁰ C.
83. EOSIN NIGROSIN
• Preparation of 10% nigrosin stain
• Nigrosin water soluble powder 10 gm + 100 ml, 2.9% sodium citrate
• Store at 4 degree temp
• Staining procedure
• A drop of eosin, four drops of nigrosin and a small drop of semen are
placed on a clean, grease free slide.
• Mix the semen first with eosin and then immediately with nigrosin stain.
• The mixture is taken on the edge of a slide and pulled across the top of
another slide leaving a smear
• Allow it to dry in air.
• 200 spermatozoa are counted under oil immersion at a magnification of
100X
87. ACROSOME INTEGRITY
• Acrosome a cap like structure on the head of the spermatozoa covers 60%
of the anterior portion of the sperm head.
• The Mammalian spermatozoon acrosome is golgi derived secratory
granule inclosed by outer and inner acrosomal membrane.
• The acrosomal matrix, which underlies the outer acrosomal membrane,
contains both structural and nonstructural (non-enzymatic and enzymatic)
components
• A double Trypan blue/Giemsa staining method was first used for sperm
evaluation by Kovács and Foote in 1992
91. BIOCHEMICAL TESTS
• pH estimation
• Methylene blue reduction test
(MBRT)
• Hypoosmotic swelling test
(HOST)
• Fructolytic index
• Oxygen uptake/ utilization test
• Pyruvate utilisation test
•Glutamic oxaloacetic
transaminase (GOT) activity
•Hyaluronidase activity
•Resazurin test
•Buffering capacity test
•Alkaline & acid phosphatase test
•Millovanov’s resistance test
92. pH estimation
Species pH
Bull 6.8
Stallion 7.4
Ram/Buck 6.8
Boar 6.8
Dog 6.7
Cat 7.4
pH paper
Indicator dyes
Bromothymol blue - 5.2-6.8
Bromo cresole purple – 6.0-7.6
Phenol red – 6.8 – 8.4.
pH of the semen affected during
Inflammatory conditions affecting the accessory
sex glands
When bulls are used excessively – Alkaline pH
Incomplete ejaculation – Alkaline pH
93. METHYLENE BLUE DYE REDUCTION TEST
• Principle that the hydrogen ions are liberated during sperm metabolism
which will reduce the blue colored methylene blue into colorless
leucomethylene blue.
• Procedure and interpretaion of reuslt
• Materials required
• Fresh semen
• Egg yolk citrate diluent
• 5 ml test tube
• Incubator
94. METHYLENE BLUE DYE REDUCTION TEST
• Methylene blue solution
• (50 mg of methylene blue in 100 ml of 2.9 per cent sodium citrate buffer)
• Liquid paraffin
• Water bath
• Procedure
• Take 0.2 ml of fresh semen and 0.8 ml of egg yolk citrate diluent in a sterile
5 ml test tube
• Add 0.1 ml methylene blue solution and mix the contents
• Place 1 cm layer of liquid paraffin
• Keep the test tube in a water bath of 46.5° C
• Observe the time taken to change in color from blue to colorless
95. METHYLENE BLUE DYE REDUCTION TEST
S.No. Time interval Classification
1 3 - 5 min Good
2 9 min Average
3 > 9 min Poor
Based on the time taken to change the color, the semen sample
is graded as
96. HYPO OSMOTIC SWELLING TEST (HOST)
• The integrity of plasma membrane is a pre-requisite for maintaining
fertility.
• Fluid transport occurs in an intact cell membrane under hypo osmotic
conditions until equilibration is reached between inside and outside the
cell.
• Due to influx of fluid there will be bulging or bending of the tail fibre
occurs.
• This phenomenon is known as "tail curling" or the sperm with curled tail is
known as "swollen sperm".
• Spermatozoa with chemically and physically intact membrane will show
tail curling under hypo osmotic conditions whereas spermatozoa with an
inactive membrane will not.
97. HYPO OSMOTIC SWELLING TEST (HOST)
• Materials required
Fresh/frozen semen
Sugar tube
Distilled water/ HOS media
Water bath
Pipette and tips
Preparation of HOS media
For 75 mosm media
Sodium citrate - 0.367 gm
Fructose- 0.675 gm
Distilled water - 100 ml
98. HYPO OSMOTIC SWELLING TEST (HOST)
• Procedure
• Take 0.9 ml of HOS media /distilled water in a sugar tube
• Keep the tube in 37 ° C water bath for 5 minutes to bring the
temperature of media to 37 ° C
• Then add 0.1 ml of semen
• Incubate the mixture at 37 ° C water bath for 30 minutes
• Place a drop of semen in a clean, grease free glass slide and put a
cover slip over it.
• Examine under phase contrast microscope to see the tail curling
99. HYPO OSMOTIC SWELLING TEST (HOST)
The fresh semen - minimum of 70% and above
Post thaw - minimum of 50% reacted sperm.
100. FRUCTOLYSIS INDEX
• Fructose, a glycolysable sugar, present in the semen
• Its level in semen is regulated by the male sex hormone testosterone
• Fructolysis in the semen is assessed by measuring the disappearance of sugars
and accumulation of lactic acid by a constant number of spermatozoa in a
specific time and under specified conditions.
• Mann (1948) for the first time proposed fructolysis as an index for evaluating
the activity of semen.
• Fructolytic index is defined as the amount of fructose utilized by
109 spermatozoa in one hour at 370C.
101. Oxygen uptake/utilization test
• Principle – Metabolic activity of sperm is measured in terms of oxygen
uptake.
• Values represented as R.Q. (Z)
• Zo2 = volume of CO2 produced by the spermatozoa per unit of time
divided by the volume of O2 consumed in the same unit of time.
• The ZO2 value of bull semen is to be 21
• This test indicates activity and live percentage of spermatozoa.
102. Pyruvate utilisation test
• Principle – Based on oxygen utilization in presence of pyruvate and
associated compounds
• First started by Melrose during 1952.
• Oxygen consumption rate after the addition of pyruvate and pyruvate plus 2,
4 di nitrophenol.
• Flouride added - reduce the exogenous metaboloism to a low level.
• When added with pyruvate the oxygen consumption in high fertile bull semen
and low fertile bull semen is increased but on addition of 2,4 di nitrophenol
the oxygen uptake is increasing two fold in high fertile bulls but not in low
fertile bulls.
• The oxygen uptake was measured with the help of monometric equipment.
103. Presence of certain Enzymes and Hormones
• GLUTAMIC OXALOACETIC TRANSAMINASE (GOT) ACTIVITY
• Indicates the damage to spermatozoa
• HYALURONIDASE ACTIVITY - indicate acrosomal damage.
• RESAZURIN TEST - indicator of metabolic activity of semen.
Resazurin
• Semen +Resazurin (Blue) Pink
Dehydrogenase
1 minute
CL
4 minute
ALKALINE & ACID PHOSPHATASE TEST
Presence indicated - functional state of accessory sex glands and
metabolic activity of spermatozoa.
Dihydroresurufin
Resurufin
104. Millovanov’s resistance test
• Evaluates ability of spermatozoa to withstand the action of 1 per cent
Sodium chloride solution.
• Denoted by – R
• The milliliter of 1 % Sodium chloride solution required to stop the
progressive motility of all spermatozoa in 0.02 ml of semen.
• Good quality semen - ‘R’ value to be not less than 5000.
105. Cold shock resistance test
• This test is done to assess the fertilizability/preservability/storage ability (5°C)
of the semen of a particular bull.
• The spermatozoa are subjected to unfavourable condition such as cold (0°C).
• The percentage of resistant live sperm is counted after cold shock.
• Procedure
• Take one ml of freshly collected in a test tube and place it in a beaker
containing crushed ice (0⁰C) for 10 minutes.
• Conduct Viability by eosin-nigrosin
106. BIOLOGICAL TEST
• Zona-free hamster ova penetration bioassay - in vitro fertility assessment
of spermatozoa.
• The hamster egg penetration is a heterologous sperm penetration test.
• hamster egg will allow the other species sperms after the zona removal by
enzymatic digestion.
• ZP - removed by treating the oocytes with freshly prepared 0.1% trypsin
in mBWW medium for 30 to 60 seconds.
108. Semen Extender
• Semen Extender or diluent is a chemical medium used for preservation,
extension and protection of sperm cells against various shocks during
processing, storage and transportation used for artificial insemination.
• Characteristics of good extender
• Isotonic (280- 310 m Osm) to seminal plasma
• Buffering capacity (regulate pH)
• Cold shock protection
• Protection from cryo-damage/cryoinjury
• Energy source (sperm metabolism)
• Control microbial contamination
109. Various components used in semen extender
1. Egg yolk
• It is non-penetrating cryoprotectant
(Cold shock protection)
• Contains
Phosphatidylcholine (lecithin)
Phosholipids
Lipid extracts
Lipoprotein fractions
Specific lipoproteins
Provides protection from cold shock
110. Egg yolk
• 20 percent Egg yolk is still used as a standard as it is cheap and readily
available
• Philips in 1939, was the first to report egg yolk use in the diluents
• In egg yolk, Low Density Lipids (LDL) abundance is considered a main cryo-
protective agent
• The LDLs are composed of 17-60 nm spherical molecules
• The LDL contains 11-17% proteins and 83-89% lipids
• Egg yolk increased post thaw motility by solubilizing the cell membrane
lipids and binds to the sperm
• But such a high amount increased the risk of microbial contamination
resulting in metritis and transboundary diseases
111. Semen
Spermatozoa
Seminal plasma
BSP Proteins (A1, A3, A5)
Sperm Membrane+ BSP
Cholesterol efflux
membrane destablisation
capacitation and AR
Good for NI Bad for cryopreservation
LDL
112. Milk sources
• Lactose
• Cannot diffuse across the cell plasma membrane, thus helps in creating
osmotic pressure
• Major protecting agent is casein micelle
• Minor – Whey protein (α lactalbumin, β lactoglobulin) and lactose
• Milk protein binds with 50-80 % BSP Proteins
• Protein-protein interaction
113. Soy-lecithin
• Replacement of animal protein source with plant protein sources
• To avoid resulting diseases and keep the biosecurity from trans-boundary
diseases
• Lecithin (present in different cryoprotectants) protects the plasma
membrane by restoring phospholipid that is lost due to heat and protects
the cell viability
• Soy-lecithin is a very good alternative to phospholipids present in egg yolk
for semen cryopreservation
• Commercially available - AndroMed, Biociphos
114. Various components used in semen extender
• Ions for maintaining osmolarity
• Zwitter ionic buffers
• Amino acids
• α-keto acids
• Combination of salts and carbohydrates are added in extenders
• Buffer
• Hydroxymethyle aminomethane (Tris) and citric acid are very common
used buffers in various types of diluents used for ruminant semen.
• Tris containing egg yolk glycerol extender was developed in 1963
115. Antimicrobials
• The most common contaminants are Gram positive bacteria along with E.
coli and Salmonella spp
• Cornel extender was the first standard diluent to have Penicillin G,
Streptomycinand Polymixim-B
• Penicillin and streptomycin are still used at the rate of one gram per liter
Energy source
Fructose
116. Glycerol
• Glycerol is classified as a penetrating cryoprotectant and most common
used for dilution of semen worldwide
• Glycerol cryo-protective effect was accidentally found by Polge and co-
workers in 1949
• Glycerol as conventional cryoprotectant prevents the intracellular
crystallization
117. Classification of semen extenders
1. Ambient temperature – 2.5x106 motile spermatozoa
IVT (Illini variable temperature)
Coconut milk extender
Self carbonating extender e.g. (CUE) Conell university extender & Caprogen
extender
2. Chilling/refrigeration temperature extender - 10x106 motile sperm
• EYC (egg yolk citrate)
• EYP (egg yolk phosphate)
• TRIS extender
• Skimmed milk extender
• Whole milk extender
• D2 diluent
3. Cryopreservation Temp. - 196
Glycerolated EYC
Glycerolated EY TRIS
119. Methods of semen collection
Collection from vagina (Spoon/Sponge method)
Massage technique
Artificial vagina method
Electroejaculation method
Digital manipulation method
120. Vaginal method ( Spoon or Sponge technique)
• The simplest and earliest method of semen collection was from vagina.
• Bull is allowed to mount the cow/dummy in naturally and the semen is
collected from vagina by means of a spoon or Sponge or syringe with a
long nozzle.
121. Disadvantages of spoon method
1. Unsatisfactory semen quality
• Selectively small volume of semen is mixed with large volume of vaginal
mucus.
• To avoid this female out of oestrus can be used.
• Later, Breeder’s bag and urethral fistular methods of semen collection
were used.
2. Non-scientific method.
3. Danger of contamination and disease transmission.
4. Keeping quality of the semen will be very poor.
122. Ampullary Massage technique
• Here semen is collected by massaging the ampullae and seminal vesicle
through the rectum.
• This method was reported as early as 1925 by Case and late well described
by Miller and Evans (1934).
• Procedure
• Before starting semen collection by massage method ensure that the bull
has not ejaculated either by natural service or by some other method for
two to three days to ensure that ampulla is full of spermatozoa.
• Secure the bull in a suitable stanchion & The preputial hair should be
clipped
123. Ampullary Massage technique
• By tapping the preputial sheath stimulate the urination and empty the
bladder
• Properly wash the preputial sheath with warm water and carefully dry it
with a clean towel
• Perform back-racking
• The operator can carefully massage the seminal vesicle, prostate, cowper’s
gland to stimulate some secretion of seminal fluid to rinse the bull’s
urethra.
• The seminal vesicles are gently massaged for few times with fingers by
backward and downward strokes towards urethra and a cloudy fluid is
expelled.
124. Ampullary Massage technique
• Then the ampullae is massaged one by one by gentle, slow and rhythmic
manner.
• The ampullae is squeezed/stripped by pressing over floor of the pelvis and
the pelvic urethra may be massaged.
• The massage can be done for 5 minutes. After the massage of the
ampullae the S-curve of the penis should be straightened to allow the
escape of semen.
• An assistant holding a glass funnel fitted to a collection vial directly
beneath the bull’s sheath can collect the semen during massaging.
• The quantity of fluid collected from seminal vesicle ranged from .5 to 21
ml and ampullae ranged from .5 to 23 ml.
125. Ampullary Massage technique
• Advantages
• Semen can be collected from bulls, which are physically incapable of
mounting due to less libido, lameness or fracture .
• To collect semen from bulls that are refusing to artificial vagina.
• To collect from bulls those are incapable of erection.
• Disadvantages
• Chances of contamination by urine or sand.
• Seminal vesicles secretion leads to unbalanced semen components than
ejaculated semen.
• Poor concentration and poor keeping quality.
• Requires skill on the past of operator or training on the past of bull.
• Excessive massage will cause inflammation of the organs.
126. Electroejaculator method
• Electrical stimulation of ampullae and seminal vesicle is also a
method of collecting semen from bulls.
• This method was used by Gunn in 1936 for rams.
• Later it was modified by Rowson and Murdock, Marden, Dzuik for
collection of semen in bulls.
• Thibault et al. (1948) successfully used this for bulls.
127. Electroejaculator method
• Procedure
• The animal is restrained in a trevis
• The preputial hairs may be clipped, washed and dried
• Teasing or sexually stimulating the bull before applying electro ejaculate
may help to inform the quality and quantity of semen
• After cleaning the rectum the glove hand with the electrodes is inserted
and passed forward upto 30 cm and the electrodes can be pressed down
on the floor of the rectum directly over the two ampullae between the
two diverting seminal vesicles
128. Electroejaculator method
• An alternate current is passed starting at 5 voltage and return back to zero
every 5 to 10 seconds.
• This is gradually increased up to 15 volts. Generally the erection and
ejaculation occurs at 10-15 volts when 0.5 to 1 ampere current is flowing.
• The semen is collected in a clean sterile glass with a glass funnel.
• The first portion generally consists of merely the watery secretion of the
accessory glands and can be discarded.
• The portion is usually containing semen is collected and used.
129. Electroejaculator method
• Advantages
• Semen can be collected from males that refuse to donate semen in artificial vagina, lowered sex
libido or when injuries or deformities make this impossible.
• Helps to collect the semen from bulls that are not able to mount
• Bulls that are recumbent but having good pedigree can be collected by this method
• Disadvantages
• The technique may be painful to bulls. It can not be practiced frequently, since frequent use of this
method may affect the health status of the bulls.
• The quality of semen collected by electro ejaculation cannot be compared with semen obtained by
AV method because the non accessory sex gland secretion.
• The semen collected by this method is usually more in volume and less in sperm concentration. The
contamination of semen will also be more.
130. Artificial vagina (AV) method
• The AV provides the most satisfactory method of collecting semen from
bulls.
• This is the most widely used and preferred method for routine semen
collection.
• This method has several advantages over other methods and a normal,
clear ejaculate can be collected.
• Artificial vagina is very convenient and is more near to the natural service
i.e. it contributes the threshold of warmth, pressure and friction
simulating the normal characters of female reproductive passage.
131. Artificial vagina (AV) method
• Limitations
• Requires trained/skilled personnel for collection of semen.
• Chances of injury by furious bulls
• Development of artificial vagina
• Russians (Kumorov and Nagev, 1932) designed the first artificial vagina.
• The Cornell University workers developed Cornell Model
• “Danish model” was developed in US
• In tropical countries the “short model” was developed
132. Russian model of Artificial Vagina
• Made of rigid rubber cylinder
• 60 cm long and 5.5 cm inner diameter.
• The inner side of rubber cylinder was covered by a thin walled rubber liner,
the ends was turned back over the outer cylinder to form water tight jacket
• The jacket was filled with hot water enough to bring the inside of AV to also
body temperature.
133. Danish model of artificial Vagina
• Danish model is commonly used for semen collection in united states.
• Length – 40.70 cm
• Outer diameter – 6.25 cm
• Inner diameter – 5.7 cm
• The above mentioned dimensions may be lowered for smaller bulls in order to
collect the ejaculate near the collecting receptacle
134. Cornell model of Artificial Vagina
• Developed by scientists of Cornell university
• Length – 71.20 cm
• Outer diameter – 6.25 cm
• Inner diameter – 5.7 cm
• This model overcome the advantage of Danish model by covering the
collecting receptacle and exposing to outside environment
135. Disadvantage of AV
• Semen receptacle is exposed to light and ambient temperature
• Damage of cold shock to spermatozoa during collection in cold weather.
• Cornell model overcome the advantage of Danish model by covering the
collecting receptacle and exposing to outside environment
138. Inner liner Inner liner/ latex liner
Rough neoprene liner
Rough latex liner
Smooth latex liner
Parts of Artificial Vagina
139. Preparation of AV
• The parts AV should be sterilized before it is used for semen collection to
avoid the contamination and disease transmission.
• After sterilization all parts should be dried and stored in a dust free
cabinet or incubator.
• The inner liner should be inserted into the AV and both the ends are
turned back over the ends of the cylinder
• Rubber bands may be attached on both the ends so as to form a jacket
between the liner and rubber cylinder.
140. Preparation of AV
• The director cone is fastened over one end of the AV and the glass semen
collecting vial is attached to the smaller end of the cone.
• After assembling AV, half the jacket is filled with warm water of about 65-
70 ⁰ C is poured to get the final temperature of 42.5 to 45 ⁰ C inside the
AV.
• The remaining half of the jacket is filled with air. The temperature of AV
may vary depending upon the season, time semen collection and air
temperature. (Click here to view picture).
• The insulation bag is used in colder or hotter environment to avoid
cold/heat shock to the spermatozoa.
141. Preparation of AV
• The insulation bag is applied to cover the director cone and
collection vial. (Click here to view picture)
• After assembling the AV, sterile lubricating jelly is applied for 3 to 5
inches of the interior of liner.
• The temperature of the AV should be checked by the semen
collector before taking it for semen collection. (Click here to view
icture)
• Excessive lubrication will cause the contamination over semen
samples by carrying the jelly through the AV over the bull’s pelvis.
142. Lubricants
K.Y
. Jelly
Sterile white Vaseline jelly
White mineral oil (Sterile)
Tragacanth gum (3 gm Tragacanth, 5 ml
glycerine and 50 ml distilled water)
143. Preparation of bull
• Semen collection is done in the early morning.
• The scheduled bulls will be carried from their paddocks to
washing area.
• The animals should be washed to remove all the dirt.
• The bulls should be tied with bull apron and the prepuce
should be cleaned with normal saline
• The bulls are allowed to watch the other bulls mounting
which will stimulate the bull.
• The forward and backward movement of dummy will also
stimulate the bull.
• The protrusion of penis from prepuce will indicate the bull is
ready to mount.
147. Semen collection
• The teaser should be restrained in a trevis.
• The dummy also used occasionally for semen collection
• 2 to 3 false mountings are given
• The right handed operator can approach the bull from right side by
holding the AV on his right hand
• When the bull mounts and ready to make the thrust, the operator
using the left hand drawn the penis from sheath and directs it into
the AV which is held at an angle that it is in have with the testis.
148. Semen collection
• During the entire process the testis itself should not be touched as it will
leads to refusal to service.
• The AV should be directed in such a way that the semen should be
ejaculated in director cone or directly on collection vial
• After collecting the semen the collection vial is disconnected immediately,
closed with clean stopper / sterile aluminum, plastered with details of
bulls, placed in a beaker containing water of about 34 ⁰ C and examined as
soon as possible
149. Prepared by-
Dr. S. K. Sheetal
Assistant Professor cum Jr.Scientist Department of
Veterinary Gynaecology and Obstetrics,
Bihar Veterinary College, BiharAnimal Sciences
University, Patna-800014
1
150. INTRODUCTION
2
• The Study of patho-physiology of male
reproductive system is known as Andrology.
• Functional anatomy of male genitalia is a
basic point to understand andrology.
• The value of the bull is more than half of the
herd in cattle breeding
152. 4
• Reproductive system = Mesodermal Origin
• In early embryonic life Nephric & Genital
regions formed from mesodermal tissue
Finally modified into Urogenital Sinus
• Cranial portion of nephric region
Pronephros
• Caudal to pronephros mesonephros
(Wolffian body) and mesonephric duct
(Wolffian duct)
153. 5
• Metanephros (Kidney) 3rd excretary organ
develop from outgrowth of mesonephric duct.
• Pronephros degenerate
• Mesonephros degenerate
• Mesonephric tubules persist
• Mesonephric duct persist and utilized in
males for transport of sperm from testes to
pelvic urethra.
154. 6
Bulging structure from mesonephros
Genital ridge/ Gonadal ridge/ Urogenital
ridge/ Indifferent gonad/ Undifferentiated
gonad/ primitive gonad/ primordial gonad
Position medial to the mesonephros
Appearance of genital ridge on mesonephros
=
Human/ Bull = 28 days
Horse = 27 days
Boar = 26 days
Dog/Sheep = 24 days
155. • Origin of primary germ cells extra gonaldal
•
• Wall of Yolk sac endoderm in the region of
hind gut.
• Migration of primordial germ cells =
• Amoeboid movement (Diapedesis)
• Chemotaxis
Secondary sex cord Medula Testis (Male)
Primary sex cord Cortex Ovary (Female)
7
156. Origin of internal ducts
8
• Embryonic stage double set of sex ducts in
vertebrate
• During differentiation only one duct system
persist
• 1. Mesonephric duct (Wolffian duct) = Male
• 2. Paramesonephric duct (Mullerian duct) =
Female
157. Testis
• Sertoli cell formation Male
• Other name Sustentacular/Nurse cell
derived from mesonephric tubule epithelial cells
•
• Secrete AMH (Anti Mullerian Hormone) or
MIH (Mullerian Inhibiting Hormone)
Inhibit the formation of Mullerian duct
(Paramesonephric duct)
9
158. • Interstitial cells (Leydig Cells) mesodermal
mesenchymal cells
Secretes Testesterone
• Seminiferous tubule remain solid until just
before puberty.
• Initial appearance of tunica albuginea 1st
histological evidence of testicular
development (7th week)
10
159. Mesonephric duct (Wolffian duct) =
Male
11
• Epididymis
• Ductus deferens or vas deferens
• Ampulla
• Seminal vesicle
160. Paramesonephric duct (Mullerian
duct) = Female
• Oviduct
• Uterine horn
• Uterine body
• Cervix
• Cranial 2/3 Vagina
• Rest part of vagina formed by Urogenital sinus
Prostate & bulbourethral gland
12
165. Male Reproductive Organs:
17
Scrotum
↓
Two testes or testicle
↓
Duct System
(Epididymis, Vas deference, Ampulla)
↓
Accessory sex glands
(Seminal Vesicle, Prostate and Bulbo-urethral Gland)
↓
Penis and prepuce
167. SCROTUM AND TESTES
19
• Scrotum-
• Cutaneous pouch derived from skin and fascia
• House of testicle.
• Location:-
• In between thigh or inguinal region in all
domestic animals except boar and cat.
• Boar and Cat caudal to thigh or perennial
region.
• Testis fixed to the scrotum scrotal ligament
attached to caudal end
173. Layers of tissues between scrotal skin
and testis proper
25
1. Tunica dartos layer:
Present under the scrotal skin
Composed of smooth muscle fibres, fibrous
and elastic connective tissue.
Surrounds both the testes.
Forms medial septum w/n two testes
2. Loose connective tissue layer
Present under tunica dartos layer
174. 26
3. Vaginal process layer:
Present under loose connective tissue layer
It is an extension of peritoneum passing
through abdominal wall at the inguinal canal.
Composed of a) Tunica vaginalis communis :
corresponds to parietal peritoneum of
abdominal cavity.
b) Tunica vaginalis propria: corresponds to
visceral layer of peritoneum of abdominal
cavity
175. 27
• 4. Tunica albuginea layer:
Present beneath the visceral layer of the
vaginal process (tunica vaginalis propria)
Tough layer composed of fibromuscular tissue.
Extensions of tunica albuginea penetrate the
testicular parenchyma to join at medistinum.
Fibrous septa divides parenchyma into
lobules
Lobules contains seminiferous tubules (75%
of testicular mass)
176. DESCENT OF THE TESTICLE
• TESTIS originates near caudal pole of kidney
• Sink through inguinal canal
Scrotum
Inguinal ligament, diaphragmatic ligament &
mesorchium Suspend the testicle
Testicular descend passive
Testis governs its own decends
Hormonally controlled from testis or adrenals
28
177. • Testicular descent gubernaculum testis
• Regressed & Decrease in size after descend
• Forms Ligamentum testis & scrotal ligament
descend of testis completed by birth or soon after
29
178. 30
Period of testicular descend
• Bull= 106 days of GP
• Ram & Boar = 70 days of GP
• Horse = at or near birth
• Dog = 3-4 days postnatally
179. Cryptorchidism
31
• One or both testes are retained either in the
abdominal cavity or in the inguinal canal.
• High flanker: the animal in which the testicle
descends in to the inguinal canal but not into
the scrotum is called “high flanker”.
• Unilateral or bilateral crypotchidism
• Unilateral spermatogenesis from one
normal testis
• Bilateral no spermatogenesis, sterile.
180. 32
Length of the seminiferous tubules in
different species
• Bull = 5 km
• Ram = 4 km
• Boar = 6 km
• Dog = 150 m
• Cat = 25 m
181. Passage of spermatozoa
33
• Seminiferous tubules Tubuli recti (straight
tubule) Rete testes Efferent tubules (6-24
in number) Epididymis Vas deferens
Ampulla Urethra
182. Component of spermatic cord
34
1. Internal spermatic artery
2. Internal spermatic vein
3. Vas deferens
4. Autonomic nerve from renal and caudal
mesenteric plexus.
5. Lymphatic vessels
6. Internal cremaster muscles
7. Tunica vaginalis propria.
183. Shape, colour of parenchyma and
measurement of testis
Horse Bull Ram Boar Dog Cat
shape Oval Elongated
/Oval
Elongated
/ Oval
Elliptical Round to
Oval
Round to
Oval
parenchy
ma
Reddish
gray
Yellow Creamy
white
Grayish to
dark red
Reddish Reddish
Measure
ment (cm)
11 x 6 x 4 14 x 7 x 7 10 x 6 x 6 13 x 7 x 7 1 x 1.2 to
4x 2.5
1.2 x 0.7
to 2 x 1.5
Weight
(gm)
200-300 200-300 200-300 150-200 7-15 ------
Plane Horizontal Vertical Vertical Oblique Oblique Oblique
35
184. Thermoregulation of testes
36
1. Lack of subcutaneous fat in scrotal skin.
2. Rich supply of sweat gland in scrotal skin.
3. Cremaster and Dartos muscle :-
Contract – during cold weather (close to body)
Relax – during summer (away from body)
4. Pampiniform plexus of the testicular vein –
arterial blood supply of the testis is cooled
down by pampiniform plexus of the testicular
vein
185. Functions of the testes
37
1. Production of testosterone (endocrine
function)
2. Production of the spermatozoa (exocrine
function)
3. Blood testes barrier protect the germinal
epithelium from immunological damages.
** Bull produces 12-17 million spermatozoa per
gram of testicular tissue daily
186. Blood testes barrier (BTB)
38
• Blood testes barrier is a physical barrier
between blood or lymph vessels and the
lumen of the seminiferous tubule of the
testes.
• Adjacent Sertoli cells (or sustentacular cells)
joined together by tight junction through
basal cytoplasmic processes over
spermatogonia.
• These tight junction form blood testes barrier.
188. 40
It divides the seminiferous epithelium into the
basal and the apical (adluminal)
compartments.
The blood-testis barrier is one of the tightest
tissue barriers in the mammalian body.
Blood testes barrier prevent entry of harmful
substances from blood affecting developing
sperms.
It also prevents sperm related proteins to
enter circulation.
189. 41
• Basal compartment:- spermatogonial renewal
and differentiation and cell cycle progression
up to the preleptotene spermatocyte stage
take place outside of the BTB.
• Apical compartment:- Meiosis I and II,
spermiogenesis, and spermiation all take place
in a specialized microenvironment behind the
BTB
190. 42
EPIDIDYMIS
• Coiled tube closely attached to the testis
• More firm in consistency than testis
3 different parts:
1.Head (caput) broad, covers 1/3rd proximal
end of testis
2. Body (Corpus) narrow, running towards distal
end of testis
3. Tail (Cauda) enlarged end at distal pole of the
testis.
continue with the vas deferens.
192. 44
• Histologically two prominent layers
• 1. circular muscle fibers layer
• 2. Pseudostratified columnar cell layer
• Based on histology -- 3 Segment
1. Proximal segment ciliated cells (kinocilia), beating
outward
almost no lumen
medusa (deattached ciliated cells) formation
2. Middle segment wide lumen, cilia are not so
straight
3. Terminal segment lumen very wide, short cilia
packed with spermatozoa
193. Functions of the epididymis
45
1. Absorption:
testicular fluids >>> semen ejaculates
Ex. Ram’s testicular fluid 60 ml, semen ejaculate 1
ml
Epithelial cells of cauda epididymis
↓
Active absorption of fluid
↓
highly concentrated spermatozoa
194. 46
2. Secretion:
Epididymal cells,
Maitain viability of spermatozoa.
3. Maturation:
Occurs during storage period
migration of the cytoplasmic droplet from the
neck of spermatozoa to the distal end of
middle piece.
increased capacity for motility & ferlilizing
ability.
196. 48
4. Transportation:
Due to ciliated epithelium.
Action of peristaltic waves of muscles fibers.
Duration of epididymal journey:
Ram = 13-15 days
stallion = 8-11 days
Bull = 10 days
Boar = 9-12 days
5. Storage:
Cauda epididymis stores approx. 50% sperms
Two epididymis stores up to 3-4 days
production of spermatozoa.
Spematozoa remains in quiescent metabolic
stage in epididymis
197. VAS DEFERENS OR DUCTUS DEFERENS
49
• Cauda epididymis pelvic urethra.
• Thick muscular walls and small lumen.
• Convoluted near cauda epididymis and run
parallel to corpus epididymis.
• Pass through inguinal canal into abdominal
cavity along with components of spermatic
cord.
• Then open into pelvic urethra
198. 50
• Vas deferens 3 mm thick in bull
6 mm in stallion
•
• Elongated terminal part of vas deferens is
called Ampulla.
• Length = 10-12 cm, diameter = 1 to 1.5 cm
• Dog & cat ampulla absent
• The opening of ampulla in the cranial portion
of the pelvic urethra through a rounded
prominance is k/a “Colliculus seminalis”.
199. ACCESSARY SEX GLANDS
1. Vasicular glands or Seminal vesicles :
Paired gland with lobulations
Located on the pelvic floor cranial & lateral
to the ampullae.
Open in the pelvic urethra near opening of
ampullae (colliculus seminalis) or both
shares common ejaculatory duct
Branched tubular secretary glands
Add volume, nutrition and buffers
200. Secretion of about 50% of total semen ejaculate
Secretions Alkaline in nature.
Contains Protein, fructose, ascorbic acid, citric
acid, potassium bicarbonate and enzymes.
Stallion – vasicular gland (elongated Pear
shaped sacs) gel to ejaculates.
Boar – large bag like milky and viscous fluid
secretions high Inositol & ergothionine
content.
Bull – secretion yellow high riboflavin
Absent in Dog & Cat.
201. 2. Prostate Gland :
• Bull : Location pelvic floor on or around
neck of bladder or cranial portiion of pelvic
urethra.
• Opens into pelvic urethra lateral to the
“colliculus seminalis” through many ducts.
• Two parts a) pars propria (body)
• b) pars disseminata
• Ram no body
• Dog Larger in older dogs
203. 3. Bulbourethral gland or Cowper’s
gland
• Paired gland
• Either side of pelvic urethra near the ischial
arch.
• Bull : embedded under bulbospongiosus
muscles.
• Bull/Ram/Stallion = Ovoid
• Boar = cylindrical
• Absent = Dog
204. • Bull: driblling before ejaculation secretions
of prostate & bulbourethral glands.
• Cleans urethra prior to semen ejaculations.
• Gel (rubber like white substance) formation in
boar semen
205. PENIS
• Copulatory organ of male
• Dorsal part made of corpus cavernosum penis
• Corpus cavernosum penis enclosed by thick layer of
tunica albuginea layer
• Ventral part surrounding the urethra corpus
spongiosum penis
• Bull /ram /boar sigmoid flexure (S –shaped curve)
• Sigmoid flexure :
• Post scrotal Bull , Ram
• Pre-scrotal boar
• Absent horse
206. • Retractor penis muscles attached to the distal
end of sigmoid flexure.
• Help in penile erection by compression and
pumping action
• Bull: penis length = 90 cm, diameter = 4-5 cm
• Glans penis = 7.5-12.5 cm long pointed
and twisted
• Penis of bull = Fibroelastic (less erectile tissue)
207. • Stallion:
• Penis length = 50 cm, diameter = 2.5-6.0 cm
(non-erected)
• Length = 2x on erection, glans penis 3x
• Urethral process in glans
• Shallow groove in glans called fossa glandis
urethral sinus or diverticulum
208. • Ram:
• 30 cm , 1.5-2.0 cm
• Characterized by urethral process extends
4-5cm beyond glans penis
• Boar:
• 45-55cm,
• No glans
• Terminal part twisted counter clockwise
209. Dog:
• Penile Length = 6.5-24 cm
• Two separate corpora cavernosa
• Cranial free portion of penis contains a bone
“Os penis” or Bacculum
• Glans penis : two parts
1. Bulbus glandis: proximal 1/3 part
2. pars longa glandis : distal 2/3 part
Bulbus glandis engorged with blood during
copulation
210. Cat:
Short and directed caudally & ventrally
Urethra lies dorsally in the penis
Os penis absent or short
Bulbus glandis absent
Glans penis absent
Terminal part contains numerous spines
(about 120) pointing backward reason for
cat cry during copulation
217. Incidence of Poor libido Bulls
(n=84)
8%
15%
77%
Very poor Poor Good
218. Incidence of Poor libido Bulls (< 5 yrs)
n =24
88%
4%
8%
Very Poor Poor Good
219. Incidence of Poor libido Bulls (5 -10 yrs)
n=30
13%
17%
70%
Very poor Poor Good
220. Incidence of Poor libido Bulls (>10 yrs)
n=30
7%
20%
73%
Very poor Poor Good
221. • General health
• Ophthalmic
• Olfactory
• Auditory
• Musculoskeletal
Eyes
Nose
Ear
Limbs, joints, vertebral
column
Structural soundness
Sexual arousal
222. An early growth of squamous cell
carcinoma on cornea
An invasive squamous cell carcinoma
Visual Stimulation – SEXUAL AROUSAL
Structural soundness contd…
EYES
223. Olfactory system
- Biostimulation : Pheromones
- Nose : free from foul discharge or lesions
- The respiratory rate should be normal
Auditory system
- Perfect ear : vocal sound perception
Structural soundness contd…
224. :
MUSCULOSKELETAL /
LOCOMOTOR SYSTEM
The feet should be free of
- Interdigital fibroma
- Abscess
- Foot rot
- Abnormal hoof growth
- Joint luxation
- Arthritis
- Nonspecific lameness
Vertebral Injury
Mounting with neck
Structural soundness contd…
225. Normal Sickle hock Post legged
(Postiness)
Camped behind
Structural soundness contd…
Side View
227. - Hooves of structurally sound bull : Even
- Need for hoof trimming : Faulty leg conformation
- Structurally unsound bull : libido worsens with age
- Fault in the rear legs are heritable
Structural soundness contd…
228. • Reproductive system
Reproductive system
It is examined for : any lesions / lacerations / injuries
Pendulous prepuce are more prone to injuries and prolapse
Normal Pendulous Prolapse preputial stricture
Prepuce
229. Penis : Best examined in sexually excited condition
Reproductive System
240. VOG-604:Andrology and Male Infertility
Dr. S. K. Sheetal
Assistant Professor cum Jr.Scientist Department of
Veterinary Gynaecology and Obstetrics,
Bihar Veterinary College, Bihar Animal Sciences
University, Patna-800014
1
241. Vices
2
Vices are bad behaviors of the animals.
Vices are more common in male animals
than females especially in large animals.
242. Reasons of Vices
3
Improper handling or abuse of animals.
Closely housed or confined in the dark.
Lack of -- exercise,
-- sunlight,
-- normal surroundings
No association with other animals.
Intact males are more aggressive.
243. Types of Vices
4
A. Masterbation or Onanism
B. Viciousness in males.
C. Slowness in breeding
244. A. Masterbation or Onanism
5
• It is observed in males of all species.
• Frequency of masterbation declines with
regular use of animals.
• Accumulation of smegma in the preputial
sheath irritation Masterbation
• Boars preputial diverticulum insert penis
into it ejaculation called “Balling up”
245. Management of Masterbation
6
• Stallion: regular cleaning of sheath
• Regular exercise
• Castration of pet dogs if not used for
breeding.
• Surgical removal of diverticulum.
• Separation of boars to prevent pederastry or
rectal copulation.
246. B. Viciousness in males
7
It is due to confinement and ill treatment.
Dairy breeds are more apt to be daangerous
than beef breeds.
Ill treated, teased and irritated animals
develop viciousness difficult to manage
247. Management of Viciousness
8
• Proper , intelligent handling of male animals
from very young age.
• Gegular daily handling, firm training and
exercise.
248. C. Slowness in breeding
9
• Acquired vice in male animals
-- favoured by improper training.
•
•
-- ill treatment.
-- painful accident at the time of
copulation.
• Management :
-- done by checking of the cause.
250. Introduction of Artificial Insemination
Dr. C. S. Azad
Assistant Professor cum Jn. Scientist
Veterinary Gynaecology and Obstetrics
drazadvet@gmail.com
251. Artificial Insemination
The term “Artificial Insemination,” commonly called “AI” implies the deposition
of Semen into the female reproductive tract by the use of Artificial
means (instruments) rather than by natural service involving the male.
(Tomar,1970)
Artificial Insemination means the deposition of the semen from a male into the
female genitalia during oestrus by mechanical means rather than by the direct
service of the respective male.
In natural mating, the male ejaculates semen directly into the vagina or near
the os uteri of the female. With the technique of Artificial Insemination semen
is collected into an artificial vagina exteriorly. It is evaluated for its qualities
and is extended and preserved with suitable media prior to use. The processed
semen is inseminated into the reproductive tract of receptive females.
A.I is a technique in which semen are collected from the male, processed,
stored and artificially introduced into the estrous female reproductive tract at
proper time for the purpose of conception.
252. HISTORY AND DEVELOPMENT
Year Author Development
1322 A.D. Arab Horse Breeders Inseminated Mares with Stallion’s semen.
1677 Antoni van Leeuwenhock and
Johann Hamm
Demonstrated motile cells in semen and
referred it as “Animalcules.”
1780 Lazzaro Spallanzani
(Father of Modern A.I)
A. I. in amphibians and in a bitch, he also
proved that the fertilizing power of semen
resided in the spermatozoa carried by
spermatic fluid
1786 John Hunter Narrated semen are mawkish and unpleasant
Taste - first it is insipid & later pungent and
the first discharge is bluish white in color &
creamy in consistency.
1799 John Hunter Artificial Insemination in Human beings.
1890 French veterinarian Repiquet A.I. in Horses and advised it as a means of
overcoming sterility.
1890 Sand and Stribolt Obtained 4 successful conceptions after
Artificial insemination in 8 mares.
253. Year Author Development
1909 Ivanovich Ivanoff (Russian) First man who successfully did A.I in cattle, sheep and birds
1914 Professor Amantea (Human
Physiologist)
Designed artificial vagina for dogs
1933 Walton Described the handling of semen
1934 Miller and Evans Ampullary massage technique in bulls.
1935 Gunn Electroejaculator for rams.
1936 Edward Sorensen and Jens Gylling
Holm
first co-operative artificial breeding association in Denmark
1938 E.J.Perry first co-operative artificial breeding association at New
Jersey in U.S.A.
1938 Milovanov Devised artificial vagina for bull, stallion and ram and
extenders for diluting the semen.
1938 Laplaud, Thiabault and Cassou Devised electroejaculator for bulls
1949 Polge, Smith and Parkes Cryoprotective effect of glycerol in frozen semen.
1951 Stewart Birth of first calf born from frozen semen
1952 Smith and Polge Glycerolas a cryoprotective agent and freezing of semen
at -196 ° C in liquid nitrogen
1955 Perks Cattle breeding association
First time used pellets as packaging materials
1957 American Breeders Service of Madison,
Wisconsin
Practiced the use of long distance transport of semen in dry
ice or in liquid nitrogen.
254. Indian Scenario
• 1939 Sampatt kumaran
First man who did A.I in cattle at the palace dairy herd of
Maharaja of Mysore. Inseminated Hallikar cows with semen collected
from Friesian bulls.
• 1942 P. Bhattacharya established A.I centre at IVRI, Izatnagar
• 1943 reported the birth of first buffalo calf born through A.I. at
Agricultural Institute, Allahabad.
• 1944 Four regional centers were established throughout India to
implement A.I on large scale at Izatnagar, Patna, Bangalore, Calcutta
and Montgomery (Pakistan).
• 1948 Dr. Veeramani Iyer first did A.I in Tamil Nadu at Madras
Veterinary College, Chennai
• 1961 Frozen semen technology was first introduced in India at NDRI,
Bangalore.
255. • First five year plan (April, 1951 to March, 1956) a master project the
key village scheme was launched.
To bring about rapid genetic improvement in the stock, artificial
insemination was accepted as a major activity of the scheme for the
improvement of cattle and buffaloes in the country
Under the scheme 600 key villages and 150 artificial insemination
centres were established during the period 1952 to 1956.
• Second Five-year plan (April, 1956 to March, 1961) the scope of work
has been further extended by 1957, 400 artificial insemination
centre’s were operating.
• Some private agencies or co-operative organizations dealing with
livestock have also adopted artificial insemination for breeding work.
256. Advantages of A.I.
Quick genetic improvement
Control of venereal diseases
Maximum possible use of best sires
Quick progeny testing
Economical
Correct breeding records
Quality of semen
Overcome size difference of animal
Overcome physical inability of bull
Easy transportation
Cryopreservation of semen
Early detection of undesirable genetic traits in the progeny
Good programme for small holding dairy farmers
257. Disadvantages of A.I.
Requires high standards of technical efficiency
Proper and timely detection of estrus
Proper knowledge of reproduction is required to avoid AI in
unwanted conditions.
Genetic defects may be transmitted quickly if selection of a bull is
not perfect.
High risk of inbreeding
Costly equipments are required
261. Basics of Artificial insemination
1. Anatomy of female reproductive system
Palpation of organs – structure, consistency etc
Proper place of semen deposition
Structures act as hurdle for AI
Vaginal folds
Urinary bladder
Fornix of vagina
262. Basics of Artificial insemination
2. Per-rectal palpation
Detection of heat
Palpation of genital structure i.e. cervix/ovary/follicle/CL
Palpation of abnormalities i.e. Endometritis
Rule out gestational heat.
Common malpractices
PRE without proper lubrication
Use of soap as lubricant
263. Basics of Artificial insemination
3. Preparation for insemination and hygiene
It’s simple to adopt good practices then to break bad ones
Proper restraining of animal
AI equipment must be kept at dry and dust free area
Wash perennial area with antiseptic solution
Protect loaded AI gun from direct sun light
Keep AI sheath in original packet till used
264. Basics of Artificial insemination
Checking water temperature
before thawing
Keep the goblet below neck
while taking an straw
Thawing of straw
265. Basics of Artificial insemination
Dipping the straw in warm water
(370C) for a period of 45 seconds
Wiping out water droplet by
use of tissue paper
Thawing of straw
266. Basics of Artificial insemination
Positioning of straw in AI gun
Straw cutting – Lab seal end
Loading of AI gun
267. Basics of Artificial insemination
Putting plastic sheath over AI gun
Loading of AI gun
268. Basics of Artificial insemination
Location & examination of cervix
Retraction of uterus
Examination of the ovaries
Palpation of follicles
Palpation of C.L.
273. Timing of AI – The game changer
• Optimum fertility - insemination at the later half of standing
heat
• Heat detection mechanism in a farm is optimum – follow AM-
PM rule
• Heat detection mechanism is not optimum and conception
rate is low – insemination soon after heat detection
274. Standard Method of Insemination
• Cows are inseminated at the internal cervical os or just inside
the short uterine body.
• Recto-Vaginal Method of AI
• Restrain the animal
• Perform Back racking
• Grasp the cervix through the rectum with the left hand.
• The vulval lips are opened by downwards pressure from the
arm in the rectum
• A catheter is then passed into the vagina and manipulated
through the cervix by the right hand.
275. The circular folds of vaginal mucosa are obliterated by
pushing the cervix forward.
The catheter is initially inserted pointing upwards at an
angle of about 30° to avoid entering the urethral
meatus or fossa
Catheter is then moved horizontally until it engages in
the external os of the cervix.
The left hand squeezes the anterior vagina on to the
caudally projecting external os of the cervix, thereby
obliterating the fornix of the vagina and facilitating
entry of the catheter into the cervix.
Standard Method of Insemination
276. Standard Method of Insemination
• Entry into the external os is accompanied by a characteristic
‘gritty’ sensation.
• The catheter is then introduced through the convoluted
cervical canal by manipulation of the cervix through the rectal
wall.
• One finger is placed over the internal os of the cervix so that
the tip of the catheter can be palpated as it emerges from the
cervical canal
277. Alternative Sites for Insemination
• Single horn insemination
• Technique require palpation of the ovaries to determine the site of
ovulation
• Careful straightening of the ipsilateral horn to allow the
insemination catheter to be inserted to within approximately 2 cm
of the uterotubal junction.
• Disadvantages
• Premature rupture of the follicle, perforation of the uterine wall
• Risk of polyspermic fertilisations
• Increase in the time required to perform the insemination.
278. Artificial Insemination in Sheep
Route Fresh Liquid Frozen Insemination Vol. Conc. Of Fresh Liquid
Frozen Inseminate (x
106/mL)
Vaginal 300 400 0.3–0.5 mL 2000
Intracervical 100 150 180 0.05–0.2 mL 1000
Transcervical
intrauterine
60 0.1–0.5 mL 200–400
Laparoscopic
intrauterine
20 20 20 0.05–0.10 mL/
horn
400–800
279. Artificial Insemination of Goats
• Effects of Seminal Plasma upon Storage
• Seminal plasma contain egg yolk–coagulating enzyme (EYCE)
that is secreted by the bulbourethral gland.
• The toxic interaction with egg yolk and EYCE
• EYCE coagulates egg yolk and hydrolyses lecithin to fatty acids
and spermicidal lysolecithins (via a phospholipase A)
• Bulbourethral gland secretions also have a toxic interaction
with milk.
• A 55 to 60 kDa glycoprotein lipase, originally named SBUIII
now called BUSgp60 releases oleic acid from milk triglyceride
281. Prepared by-
Dr. S. K. Sheetal
Assistant Professor cum Jr. Scientist ,
Department of Veterinary Gynaecology and Obstetrics,
Bihar Veterinary College,
Bihar Animal Sciences University, Patna
1
282. TERATOLOGY
It is the division of embryology and pathology dealing
with the abnormal development and malformations of
the
antenatal individual.
2
283. Definitions
Anomaly:
•It refers to the malformation involving only an organ or
part of the body.
Monster:
• It refers to an animal with extensive deformity.
Teratogens
Variety of environmental factors or agents causing
non- genetic anomalies or monsters.
3
285. Achondroplasia, or dwarf, “comprest” or
“bull dog calves”
5
Most common type brachycephalic “snorter” dwarf in
Herefords with’ short, broad head, bulging forehead,
malocclusion of the jaw, prognathism of mandible, pot-belly,
low viability and great susceptibility to bloat and dystocia.
Autosomal recessive defect with some modifiers.
“bulldog” calves is usually aborted about the fifth to eighth
month of gestation.
Hydramnios occurs in pregnant Dexter cattle carrying a
“bulldog” calf.
287. • It is a condition where skin fails to form.
It occurs most commonly on the legs below the knees
and hocks and on the muzzle, ears, tongue and mucous
membranes.
It has been described in Holsteins,Ayrshires, Jerseys,
Brown Swiss and Shorthorns.
Epithelioenesis Imperfecta Epitheliogenesis Imperfecta-
Closeup
Epitheliogenesis imperfecta
7
288. • Hypotrichosis congenita or alopecia is a recessive defect
characterized by degrees of hairlessness in Holsteins, Polled
Herefords.
Acroteriasis congenita or amelia and hemi melia is seen in
Holsteins and Brown Swiss and other breeds. This is
characterized by missing, shortened, deformed, or
“amputated” limbs.
8
289. 9
Muscle contractures and ankyloses, or arthrogryposis has
been reported as a recessive in Dole cattle in Norway; and a
dominant with incomplete penetrance in England.
Hydrocephalus in Herefords, Ayrshires, Holsteins and other
breeds. It is characterized by the birth of “dummy” or
“bawler” calves that are unable to nurse properly and die in
several days. The heads may be enlarged or normal in site
but section of the head and brain reveals distended
ventricles.
291. Double Muscling in Cattle
11
INHERITED AND GENERALLY NONLETHAL DEFECTS IN CATTLE
Polydactylism An autosomal dominant character with
incomplete penetrance.
Syndactylism or “mule-foot” single autosomal recessive. The
front feet of the Holstein calves were much more frequently
affected with syndactyly than the rear feet.
Muscular hypertrophy or “double” muscling is characterized by
reduced fat deposits, light bone, thin skin, and large muscles.
When fetuses are affected dystocia often occurs.
292. 12
Vestigeal tail (Holsteins, Angus, Shorthorns).
Fused teats (Guernseys, Herefords).
Supernumerary teats (all breeds).
Missing phalanges or “Creeper” calves (Swedish cattle).
Impacted premolars and “parrot-mouth” (Shorthorns) Short
spine (Norwegian cattle).
Agnathia or absence of a lower jaw (Jerseys and other
breeds) Opacity of the cornea is probably a recessive character
in Holsteins.
Dermoid cysts on cornea, (Herefords and Guernseys).
293. Non-Genetic Teratological Defects
Susceptibility
The zygote is not as susceptible to teratogens during the
period of the ovum or blastula or the period of the fetus as
it is during the period of the embryo and organogenesis,
especially the first half of that period.
Ovum
(12 days)
The period of early differentiation in the embryo or about the
time germ layers and organs are developing - Highly
susceptible.
Susceptibe
Embryo
(12-45 days)
Foetus
(45 to term)
13
294. TERATOGENIC AGENTS OR FACTORS
14
Nutritional deficiencies in the dam
Vitamin A and E, riboflavin, folic acid, pantothenic acid, niacin
and other vitamin deficiencies, minerals such as iodine and
possibly manganese, and amino acids such as tryptophane
may cause congenital defects. Hypervitaminoses A and D will
also cause anomalies.
Endocrine disturbances of the dam
Diabetes, thyroid malfunction, and large exogenous doses of
glucocorticoids, ACTH, insulin, androgens, progestagens,
estrogens, thyroxine and thiouracil will cause defects of the
embryo. Large doses of glucocorticoids in pregnant animals
at the proper stage of gestation may cause cheilo or
palatoschisis. Progestagens given during pregnancy may
cause masculinization of the genitalia of female fetuses.
295. 15
Physical factors
Reduced atmospheric pressures, hypothermia, hyperthermia and,
anoxia cause anomalies.
Radiation
X-ray or radioactive substances induces congenital defects.
Drugs or chemicals
Thalidomide, quinine, sulphonamides, tetracycline, streptomycin,
salvarsan, lead, mercury, nicotine, malathion,
tetrachloride, apholate, selenium, fluorine, cytotoxic
carbon
agents
including aminopterin in sheep, nitrogen mustard, actinomycin D,
6 mercaptopurine, azoserine, azo dye, trypan blue and other
dyes, salicylates, histamines, ergot, “Diamox,” reserpine,
phenylmercuric acid, galactose, E.D.T.A. , Veratrum californicum
and locoweeds possibly containing lathyrogens, all produce fetal
anomalies in animals under certain conditions.
296. 16
cholera in swine, feline
Infections
Blue tongue in sheep, hog
panleucopenia in cats, bovine virus diarrhea- mucosal
disease virus, and toxoplasma can cause anomalies in the
embryo.
Ageing of ova
By delaying ovulation 24-48 h was characterized by a three-
fold increase in chromosomal anomalies with a higher
incidence of embryonic death in rats. Similarly ageing of
rabbit spermtozoa before permitting them to fertilize eggs
resulted in normal fertilization but greater embryonic death
losses. Thus age affects the genes and chromosomes as does
other agents.
297. Hydrocephalus
Occurs due to an abnormal accumulation of fluid in the
cranial cavity.
Internal hydrocephalus is due to excessive fluid in the
ventricular system.
External hydrocephalus is rare and due to excessive fluid
between the brain and dura mater.
Internal and external hydrocephalus may be combined.
17
298. SCHISTOSOMUS REFLEXUS
Seen most commonly in cattle, rare sheep, goats, swine.
Marked ventral curvature of the spine so the occiput of the head
lies near the sacrum.
The body and the chest walls are bent laterally and the thorasic
and the abdominal viscera are exposed.
The pelvis is deformed, liver is abnormal in shape and cystic.
The rumen is occasionally distended with fluid.
The limbs are usually ankylosed and rigid.
In rare cases the limbs and head may be enclosed in a complete sac
of skin.
Schistosomus reflexus
In Bovine
18
299. CAMPYLORRACHIS SCOLIOSA
Campylorrachis scoliosa is a fetal monster, rarely seen in cattle
and swine.
It is characterized by a lateral curvature of the spine.
The limbs are usually deformed and ankylosed.
Lateral View Dorsal View
19
PEROSOMUS ELUMBIS
Seen occasionally in cattle and swine.
Characterized by a lack of vertebrae and spinal cord caudal to the thoracic
region.
The monster has a small, flattened, deformed pelvis with strongly ankylosed
and flexed hind limbs and atrophy of the muscles of the rear quarters.
300. PEROSOMUS HORRIDUS
Caprine Perosomus
Horridus
Perosomus horridus is a bovine fetal monster with general ankylosis and
muscle contractures.
This is due to a marked double S-shaped lateral twisting of the vertebrae.
It is characterized on external examination by a short spine.
20
301. 21
Miscellaneous anomalies due to displacement of tissues include
dermoids, and dentigerous cysts.
o Dentigerous cysts
Characterized by a displaced dental follicle containing
fluid and teeth, is seen most
commonly in the horse.
May be located beneath the ear and are called an “ear
tooth.”
o Dermoids
Seen occasionally on the cornea, third eyelid or on the
neck in cattle and other species.
In horses dermoid tumors may rarely involve the ovary
or testis, especially the retained testis.
302. AMORPHUS GLOBOSUS
Holocardius amorphus or Amorphus globosus
The general body form is unrecognizable.
It may occasionally be seen, most commonly in the cow, but also in the mare,
sheep and goat attached to the placenta of the normal monozygotic or dizygotic
twin.
These very imperfect zygotes are parasitic upon the placenta of the normal twin
and are never observed in single births.
Usually appears as a round or oval, edematous structure weighing 1/2 to 7 lbs.
Covered with skin and hair and containing connective tissue, fat and other soft
tissues and occasionally cartilage and bone.
Amorphus globosus was an imperfect zygote of dizygotic twins.
The monster composed mainly of stomach and intestinal tissues.
Fig.1: Gross
morphology of
amorphus fetus
with pigmented
skin and a few
hairs.
22
303. 23
CONJOINED TWINS
Conjoined twins in which the components or component parts are
symmetrical are called Diplopagus monsters or “Siamese” twins.
Triplopagus is extremely rare.
Conjoined twins arise from a single ovum and are monozygotic.
Occurrence: about once in 100,000 bovine births.
Most common in cattle but are seen rarely in sheep, pigs, dogs
and cats and are extremely rare in horses.
304. 24
CONJOINED TWINS
Type I: Those conjoined twins in which each component is complete or nearly so
include
Thoracopagus, sternopagus, or ziphopagus twins are joined at or near the
sternal region. The internal organs are usually duplicated. The components
are face to face.
Pygopagus monsters are connected at the sacrum and the components are
back to back.
Craniopagus twins are united at the heads. Components may be facing in the
same or in the opposite direction.
Ischiopagus fetuses are joined at the lower pelvic region and the bodies
extend in a straight line and the heads in the opposite direction.
Type II: The two components equal one another in this group but each is less
than an entire individual.
Usually associated with lateral fusion.
305. Either face may be complete or one eye of each may be fused into a
common medial orbit. Di-, tri- or tetraophathalmus and di-, tri- or
tetraotus may be present. 25
May vary from single normal individuals to those of two normal but
superficially joined individuals.
Duplication may lead to doubling of the cranial end of body while the caudal
end remains single; or the caudal part
may be doubled and the cranial part single.
Duplication can occur at both cranial and caudal ends with the middle area
of the monster remaining single.
Duplication, of the cranial part of the fetus is more common than that of the
caudal portion.
DUPLICATION IN THE CRANIAL REGION
Monocephalus
Monsters with partial duplication of the frontal region, nose and mouth
are referred to as Diprosopus or double face.
306. M h
a e referred to as Diprosopus or double face.
May vary from single normal individuals to those of two normal but
superficially joined individuals.
Duplication may lead to doubling of the cranial end of body while the caudal
end remains single; or the caudal part
may be doubled and the cranial part single.
Duplication can occur at both cranial and caudal ends with the middle area
of the monster remaining single.
Duplication, of the cranial part of the fetus is more common than that of the
caudal portion.
DUPLICATION IN THE CRANIAL REGION
Dicephalus or two heads, with distomus or monostomus occasionally isseen.
Mon ocephalus
onsters with partial duplication of the frontal region, nose and mout
r In Cow In Cow In Buffalo
Either face may be complete or one eye of each may be fused into a
common medial orbit. Di-, tri- or tetraophathalmus and di-, tri- or
tetraotus may be present. 26
308. 28
A Dicephalus dipus dibrachius monster has two fore- limbs and two hind limbs with partial
duplication of the spine and one or two tails, dicaudatus.
Dicephalus dipus tribrachius has 3 forelimbs
Dicephalus dipus tetrabrachius has 4 forelimbs.
DUPLICATION IN THE CAUDAL REGION
Duplication in the caudal region is designated as Dipygus. Tripygus is very rare.
Monocephalus tripus dibrachius has 3 rear limbs.
Monocephalus tetrapus dibrachius has 4 rear limbs.
Cephalothoracopagus has a single neck and more or less complete fusion of the heads to form
an almost single face.
Syncephalus has one face, four ears, and a single or partially doubled cerebrum.
Janiceps is a monster with two faces on opposite sides of the head.
DUPLICATION OF CRANIAL AND CAUDAL REGIONS
Duplication of both cranial and caudal regions is Dicephalus dipygus.
o Dicephalus tripus tribrachius
o Dicephalus tetrapus tetrabrachius
o Dicephalus tripus tetrabrachius, or
o Dicephalus tetrapus tribrachius
309. UNEQUAL AND ASYMMETRICAL CONJOINED
TWINS
Unequal and asymmetrical conjoined twins are composed of one very imperfect and
incomplete twin, called the parasite, dependent on the other twin, the autosite. This
is called a heteropagus monster. The autosite is nearly normal and the parasite is
attached to it as a dependent growth.
o The parasite may be attached to the visible surface of the autosite. Common
junction sites are the back, thorax, sacrum or pelvis, and in rare cases the
abdomen, head, or palate. The latter is called epignathus. Some of the smaller,
more imperfect parasites may be called teratomas.
o A parasite can rarely develop within the autosite usually in the abdominal,
thoracic, pelvic or cranial cavity, or in the spinal canal or scrotum.
o Teratomas in the abdominal cavity and between the mandibles have be2e9n
312. Introduction
⚫ Inability or reduced ability to fertilize the ovum due to pathology of testis,
epididymis and accessory sex glands is called as Impotentia Generandi.
⚫ Fertility in male is the normal functioning of the testes, accessory sex glands
and ducts to deliver sperm of normal quality and quantity.
Impotentia
Generandi
Associated with apparently
normal semen production
Associated with abnormal
semen production
Conditionscausing partial
orcompleteinability to
impregnatenorm
al cycling
females
313. Associated with apparently normal semen
production
⚫ Bulls infected with brucellosis, vibriosis, trichomoniasis, IBR-IPV virus,
and mycoplasma may produce normal semen.
⚫ Intrauterine insemination of brucella infected semen usually results in
infertility.
⚫ Many infertile bulls had lower DNA content of the spermatozoon nucleus
than the fertile bulls.
