Semen Collection Methods for AI

• Semen collection retrieves sperm from ejaculation for Artificial Insemination (AI).
• High-quality sperm in large volumes is preferred for artificial insemination.
• Sperm are sensitive to temperature and sunlight.

Semen Collection Methods

• Artificial vagina, electroejaculator, massage, post-coitus collection, teasing, and Semen Epididymal Recovery (SER) are all semen collection methods.

Artificial Vagina

• Aims to replicate natural physiological conditions of a female's vagina in shape, temperature and texture.
• Bulls mount a teaser or dummy cow, penis is guided into the artificial vagina to collect semen.
• Effective for large livestock like cattle, horses, and buffalo, as well as small livestock.
• Consists of an outer liner, inner liner, a rubber band, a cone and collection tube.
• Outer liner adjusts in length and has opening for warm water and air.
• Inner liner is longer than the outer with end folded outward.
• Rubber band locks liners.
• Cone prevents semen from spilling and connects to the collection tube.
• Collection tube collects the semen.
• Protective cloth protects semen from temperature changes and sunlight.
• Assemble device; attach inner liner to outer liner, attach cone/collection tube, fill with warm water, apply lubricant.
• Prepare the holding area with bedding.
• Clean bull's abdominal area, prepuce, use towel.
• During implementation the penis is directed at a 45° angle inside.
• After ejaculation, hold the artificial vagina upright, gently shake to allow semen to flow inside collection tube.
• Rinse with cold water then warm soapy water, sterilize glassware, dry equipment.
• Requires trained bull, trained operator, adequate facilities, and a teaser.
• Advantages are high semen quality, large collected volume, high sperm concentration, low contamination risk.
• Disadvantages are requirement for trained bull, high libido, may be hard to use on wild or obese bulls, and injury.

Electroejaculator

• Uses a low-power electric current between 2 electrodes to stimulate the accessory glands.
• Electrojaculation is used when using an artificial vagina is not feasible.
• Probe goes into rectum, electrical stimulus is applied to induce erection.
• Nerves originate from the lumbosacral plexus.
• Prostate stimulation induces erection; ampulla and vesicular glands induce ejaculation.
• Erection and ejaculation are controlled by the parasympathetic and sympathetic nervous systems, respectively.
• Probe conducts electrical impulses, voltmeter displays voltage, battery is the source.
• Ensure electrodes are clean, check if the device is functioning, restrain animal, insert probe into rectum.
• Start stimulation by raising voltage gradually.
• Stop stimulation when animal shows impending ejaculation, which indicated by pushing hips forward.
• Animals may urinate post ejaculation, which may contaminate semen.
• Do not exceed 15V, may cause injury.
• For lumbar region: Improper placement will result in no erection and thick ejaculate.
• Can be used on animals with paralysis or impotence.
• Not as good as artificial vagina, time-consuming and costly.

Massage

• Palpates the accessory glands to stimulate semen release.
• Hand goes into the rectum, massaging seminal vesicle, stroking urethra.
• Males are first stimulated using teaser.
• Preputal and rectal area must be cleaned to reduce risk of contamination.
• Downward pressure is applied on vesicles and ampulla.
• Another assistant collects semen.
• It can be used for impotent males.
• Detects abnormalities in the accessory glands.
• Suitable for males unable to mount, requires minimal equipment.
• High risk of contamination, low sperm volume, low sperm quality, trauma.

Teasing

• Used for dogs.
• Technique massages the bulbus glandis to stimulate arousal and erection.
• Erection then continues in collection tube, female can assist the process.
• Massaging the bulbus glandis of the dog's penis until erection occurs.
• Advantages are effectiveness, speed, simplicity, and minimal equipment.
• Disadvantages are contamination, and may cause penis trauma.

Post-Coitus

• Semen collected after natural mating with sterile spoon.
• Not recommended because semen becomes contaminated.
• Semen can be collected from anterior part of the vagina using long spoon.
• Quick, simple, performed without coercion.
• Easily contaminated, requires natural mating, contaminated with female reproductive tract secretions.

Semen Epididymal Recovery (SER)

• Collects semen through epididymis for deceased males with erectile dysfunction or sexual disorders.
• SER performed post castration for living animals.
• Applicable to any kind of animal.
• Used when natural mating is impossible.
• Post-mortem collection can be performed in multiple ways.
• Incision is made using a blade on cauda epididymis to release seminal fluid.
• Aspiration made to epididymis and use pipette to collect.
• Float up the cauda epididymis.
• Apply semen extender to ductus deferens. Retrograde flush the semen.
• High semen quality.
• A large portion of spermatozoa can be obtained.
• Can be used for breeding of endangered species.
• Creates many straws, good for artificial insemination.
• Disadvantages are possible contamination, semen quality can degrade if not packaged properly.

Fresh Semen Evaluation (Macroscopic)

• Should be done right after extraction, it includes volume, color, consistency, and pH levels.

Volume

• Done via conical tube to estimate density.
• Volume depends on animal age physical condition, health status, nutrition, collection procedure, and breed.
• Spermatozoa per ejaculate is more important than actual volume.
• Bull: 5-8 cc, Ram: 0.8-1.2cc, Boar: 150-200cc, Stallion: 60-100cc, Dog: 3-80cc, Rooster: 0.2-0.5cc

Color

• Is normally milky-white color, the cloudier it is the more concentrated the sperm.
• Bull has slight creamy white color, ram is creamy white, boar and stallion cloudy grayish white, rooster milky and white.
• Yellowish is due to riboflavin pigment.
• Yellowish green is presence of Pseudomonas aeruginosa bacteria.
• Dark red color is presence of blood.
• Light brown and greenish means contamination.
• Brownish means blood decomposition.
• Clumps and clots indicates an issue with accessory glands.

Consistency

• Can be done by shaking semen, the cloudier it is the higher the concentration.
• Different grades of semen and quantity; watery at <150 million, milky 150-350 million, thin creamy 400-800 million, creamy 1500-2000, thick creamy >2000.

Acidity

• Effects sperm survival rates.
• pH is checked, sperm acid is increased when the concentration is high.
• Bull: 6.4-7.8 Ram: 5.9-7.3, Boar: 7.3-7.8, Stallion: 7.2-7.8 Dog: 6.3-6.7, Rooster: 7.2-7.6.

Oder

• It naturally has a fishy oder due to pus.
• Mass movement is done by putting semen on object glass, without glass and dilution with magnification of 10x10.
• High temperatures above 50 degrees can quickly kill sperm and reduce motility.
• Poor indicated no movement, fair indicated few waves, good is thin with small waves.
• Very good is large waves with many dark sections.

Motility

• Spematozoa is straight in field of view, affected by age of sperm, biochemical differences.
• Viscosity and pH impact the sperm.
• Spermatozoa movement types include progressive, circular, reverse, vibratory, oscillatory, imotile.
• Individual quality is good at 70-100 range. Prepare saline at 37C, mix it, put on glass, use 40x10 glass to view with at least 3 fields of view, view the progressive movement levels.

Concentration

• Spermatozoa density is based on density per mm, measured with hemocytometer.
• Dilute with water or formalin, observe at 400x magnification.
• Can be classified as densum is less distance that first head, estimated at above 1000million.
• Semidensum (SD) is equal at 1-1.5 to first head.
• The results with raro (R) is that the first head is bigger than the other, estimated 200-500.
• At Oligospermia the concentration measures less than 200 ml.
• Aspermia means there is no spermatoza.