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TECHNIQUE OF ARTIFICIAL INSEMINATION IN CATTLE Prof. Dr. G.N Purohit MVSC, PhD, MNVAS Head Dept. Veterinary (Gynecology & Obstetrics) College of veterinary & Animal Science, Bikaner, Raj. India. Email: [email protected] Dr. GN Purohit - Confidential - Content & Video Not Permitted for Com...

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TECHNIQUE OF ARTIFICIAL INSEMINATION IN CATTLE Prof. Dr. G.N Purohit MVSC, PhD, MNVAS Head Dept. Veterinary (Gynecology & Obstetrics) College of veterinary & Animal Science, Bikaner, Raj. India. Email: [email protected]

Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

About Me : Professor G N Purohit, MVSc,Ph.D, MNVAS Head, Department of Veterinary Gynecology and Obstetrics, Veterinary College, Bikaner, Rajasthan, India

Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

Todays Topic – Technique of AI in cows. ■ Preparation for insemination and sanitation■ -Preparation of AI Gun ■ -Insemination technique

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Preparations for insemination and sanitation ■ Ensure that the cow to be bred is truly in heat and the semen to be used has motility. ■ Restrain the cow first and then thaw the semen. The restraint area should be familiar to the cow and free of stressful conditions. Unnecessary excitement may interfere with physiological mechanisms important to achieving a good conception rate. ■ Use proper sanitation

Insemination supplies should be kept dry and clean at all times. Breeding sheaths should be stored in the original package until used. Once the insemination device is assembled it must be protected from contamination and cold shock temperatures. Materials used to lubricate the rectum should not come in contact with the vulva region. Lubricants are generally spermicidal. Avoid using products that are irritating. Alternatively non spermicidal lubricants can be used.

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Common suggestions in preparation of AI guns using frozen semen

Avoid raising straws above Use forceps to remove Place straws in 95º F (350 -370C) the frost line water to thaw for at least 30-45 straws seconds. Use straws within 15 min

When all straws are used in Wipe straw completely dry the top goblet, remove and before placing in discard it. prewarmed A.I. gun

Cut straws ¼" below lab seal at 90º angle.

■ Locking mechanisms for sheaths of AI Guns • Split sheaths are designed for use with O-ring guns. • Non-split sheaths are designed for use with spiral or Kombicolor guns. Straw Size • Sheaths without plastic inserts are exclusively for use with ½ cc straws. • Sheaths with plastic inserts will accommodate both ¼ and ½ cc straws

AI Guns and sheaths

Sheaths are available in bulk or individual packages. Individually packed sheaths are better

Three types of A.I. guns

Slide sheath over A.I. gun

Slide sheath underneath loosened o-ring

Lock sheath and gun together with snug twist of the o-ring

Proper seal between straw and sheath is extremely important

In recent years more precise card type and other electronic thermal regulators of temperatures for thawing semen at the most accurate temperatures have become available (1-3). Similarly for accurate cutting of straws at 900 straw cutters are available (4) (3)

(1)

(2)

(4)

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■ Technique of insemination ■ The vulva region must be thoroughly wiped clean with a paper towel. ■ To avoid the possibility of entering the urethral opening on the floor of the vagina, the insemination gun should be inserted into the vulva upward at a 30 ̊ to 40 ̊ angle.

■ Place the cervix onto the insemination gun. Maintain slight forward pressure on the gun while manipulating the cervix slightly ahead of the gun.

Re-cap – What we learned ■ Ensure that the cow is in heat and restrain the cow first and then thaw the semen

■ Keep the material dry and clean, prevent contact of AI gun spermicidal lubricants ■ Avoid raising straws above frost line and use forceps to remove semen straws ■ Thaw semen at 350C to 370C for 30-45 secs

■ Cut straws straight at 900 angle ■ Split sheaths are used with O ring guns ■ Non-split sheaths are used with spiral or kombicolor guns

■ Sheaths with plastic inserts will accommodate both 0.5 cc ■ and 0.25 cc straws ■ Insert the AI gun at 300 to 400 angle in the vagina and

■ place cervix onto AI gun. Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

Contact me Prof. Dr. G.N Purohit MVSC, PhD, MNVAS Head Dept. Veterinary (Gynecology & Obstetrics) College of veterinary & Animal Science, Bikaner, Raj.India.

Email:[email protected] Linked in : linkedin.com/in/dr-govind-narayan-purohit-13a04a17 YouTube Channel link : https://www.youtube.com/user

/gnpobs61/videos

Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

KINDLY LISTEN TO THE LECTURE COMPLETELY, SHARE THE VIDEO AND SUBSCRIBE TO MY YOU TUBE CHANNEL GOVIND NARAYAN PUROHIT IF YOU LIKE THEM.

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ARTIFICIAL INSEMINATION IN CATTLE: DEFINITION, ADVANTAGES AND HISTORY Prof. Dr. G.N Purohit MVSC, PhD, MNVAS Dean Post Graduate Studies RAJUVAS and University Head Dept. Veterinary (Gynecology & Obstetrics) College of Veterinary & Animal Science, RAJUVAS, Bikaner, Raj. India. Email: [email protected]

Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

About Me : Professor G N Purohit, MVSc,Ph.D, MNVAS Head, Department of Veterinary Gynecology and Obstetrics, Veterinary College, Bikaner, Rajasthan, India

Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

Todays Topic –Advantages of AI and History ■ Definition of AI ■ Advantages/Disadvantages of AI ■ History of AI

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Artificial insemination (AI) is the process of collecting sperm cells from a male animal and manually depositing them into the reproductive tract of a female.

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ADVANTAGES OF ARTIFICIAL INSEMINATION:

Saving expenditure on bull maintenance

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Preventing spread of venereal disease Campylobacter

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Semen of worthy bulls is available even after death or physical disability

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Progeny Tested Bull semen can be used

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Many cows can be inseminated from a single ejaculate

400 straws Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

Cow unable to stand or refuses a bull can be mated

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Accurate Records can be maintained

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Due to regular evaluation of semen problematic bulls can be detected

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Disadvantages of AI ■ Requires specialized equipments and trained personnel ■ Requires estrus detection ■ Improper sanitation and insemination may reduce conception rates and increase disease spread

■ Reduction in gene pool due to use of superior sires only ■ Improperly tested bulls may increase disease spread multifold

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HISTORICAL EVENTS IN AI ( IN WORLD)

1300 A.D. Arabs performed AI, not scientifically documented but mentioned in an Arabic book. (Chief of Darfur) 167778 : Spermatozoa were first seen And described by Antoni van Leeuwenhoek and his assistant Johannes

1784 : More than 100 years later, in 1784, the first artificial insemination in a bitch was reported by the scientist Lazzaro Spallanzani (Italian physiologist, 1729-1799). This insemination resulted in the birth of three puppy’s 62 days later Spallanzani is also called father of Artificial Insemination. It is believed that Spallanzani was the first to report the effects of cooling on human sperm when he noted, in 1776, that sperm cooled by snow became motionless. Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

1899 : In 1899 the first attempts to develop practical methods for artificial insemination were described by Ilya Ivanovich Ivanoff (Russia, 1870- 1932). He was a pioneer in the selection of superior stallions multiplying their progeny through AI.

1914 : First Artificial vagina for collection of semen in dog was made by G.Amantia ( human physiologist in Rome University) and for Sheep and Goat by Prof.Fred.F.Mckenzie (USA). Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

1937 : Danish Veterinarians developed the first Recto-vaginal / Cervical fixation method of A.I. 1939-40 : Phillips and Lardy (1939) were the first to use egg yolk for protecting the Bull sperm from temperature shock upon cooling . This protection was explained by the effect of phospholipids and lipoproteins in the egg yolk. Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

Phenomenal growth of AI occurred in the 1940s in the United States. The procedures developed in the United States became established worldwide In 1938, an AI cooperative was established in New Jersey, modeled after the Danish system

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1941 : Salisbury and associates improved the media by using egg yolk with sodium citrate, permitting the use of semen at 5° C for up to three days

1949: Polge and co-workers were the first to freeze fowl and bull spermatozoa by using glycerol in the extender media, by describing its cryoprotective functions. (Most important milestone in AI) In 1951, with Steward, they reported frozen semen pregnancy. Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

1950 : In 1950 Cornell University scientists (New York) discovered the benefit of antibiotics added to semen in artificial insemination processes, protecting against possible contamination.

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1960 : Adler developed the technique of freezing the semen in straws using liquid nitrogen vapor.

1963 : Cassou developed the cotton plug consisting of PolyvinylAlcohol powder for sealing straws made of Polyvinyl Chloride. Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

1964-68 : Cassou developed semen straws of various size and volume and called them as medium French straws ( 135mm,2.8mm, 0.5ml) and mini French straws ( 135mm, 2mm, 0.25ml). 1972 : Germany developed plastic semen straws called Lanshut system or German straws or mini tube, sealed by metallic or plastic ball (65 mm. 1974 : Nishekawa develpoed technique of freezing semen in liquid Helium at – 265 degree celsius.

HISTORY OF A.I. IN INDIA 1939 : A I was done for the first time by Sampat Kumaran at Palace dairy farm Mysore. ■ He used HF semen to cross Hallikar cows and got 33 positive results. 1942 : A pilot project under Dr .P.Bhattacharya was started to study the feasibility of the in India. ■ This team recommended the AI to be feasible in indian conditionand

■ can be used at a mass scale. ( Pan India cross breeding began ) ■ 1943: First Buffalo calf was born through AI at Allahabad Agricultural Institute. ■ 1951-61: AI was given prominence in the breed improvement program in the first and second five year plan , key village centers were formed for this purpose.

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Re-cap – What we learned ■ The advantages of AI include wider use of good bulls and prevention of venereal diseases, opportunities for breeders to select and use of semen even after death of the bull. ■ A major disadvantage of AI is the reduction in gene pool and requirement of specific instruments and trained personnel. ■ Antony Van Leuvanhock 1677 were the first to visualize spermatozoa

■ Lazarro Spallanzani were the first to perform AI in a bitch (1784) ■ Amantia (1914) developed the first artificial vagina for dog ■ Polge discovered glycerol for semen cryopreservation (1949)

■ Adler (1960) developed the method of semen freezing in liquid nitrogen. ■ Cassou (1964) developed the french medium and French mini straws. Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

Contact me Prof. Dr. G.N Purohit MVSC, PhD, MNVAS Head Dept. Veterinary (Gynecology & Obstetrics) College of veterinary & Animal Science, Bikaner, Raj.India.

Email:[email protected] Linked in : linkedin.com/in/dr-govind-narayan-purohit-13a04a17 YouTube Channel link : https://www.youtube.com/user

/gnpobs61/videos

Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

KINDLY LISTEN TO THE LECTURE COMPLETELY, SHARE THE VIDEO AND SUBSCRIBE TO MY YOU TUBE CHANNEL GOVIND NARAYAN PUROHIT IF YOU LIKE THEM.

Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

FUNCTIONAL ANATOMY AND PHYSIOLOGY OF MALE DOMESTCIC ANIMALS- BASIC STRUCTURES AND SCROTUM Prof. Dr. G.N Purohit MVSC, PhD, MNVAS Head Dept. Veterinary (Gynecology & Obstetrics) College of veterinary & Animal Science, Bikaner, Raj. India. Email: [email protected]

Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

About Me : Professor G N Purohit, MVSc,Ph.D, MNVAS Dean Post Graduate Studies and University Head, Department of Veterinary Gynecology and Obstetrics, Veterinary College, Bikaner, Rajasthan, India

Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

Todays Topic –Male Reproductive Organs ■ Testis and other structures ■ Testicular descent ■ Scrotum

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Embryonic development of the male reproductive tract

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The male reproductive tract develops from the Wolffian duct and body or mesonephri c tubules.

The male reproductive organs consist of two testes which are present in the scrotum , ducts, accessory sex glands , penis and prepuce.

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The Testes- Function

Spermatozoa- Male gamete

Male sex hormone (androgens). Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

SCROTUM The scrotum is a bi-lobed sac or pouch (derived from the skin and fascia) and is the external part of male reproductive system which encloses the testis.

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Structures help in fluid portion of semen and transport of semen from testes to the vagina of female

The ducts that transport the sperms from testes include

Copulatory organ Penis enclosed in the prepuce

Accessory sex glands Prostrate

Epididymis

Seminal Vesicles Ductus Deferens

Bulbourethral glands

Urethra Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

Testicular descent Testis during the fetal life is located in the abdominal cavity, they pass through the abdominal inguinal canal, and inside the scrotum

Enlargement of the gubernaculum and its disappearance play an important role in testicular descent Intra-abdominal pressure in the fetus also determines testis descent

Androgen from fetal testis favors testis descent

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■ Time of testicular descent – Stallion - 9 -11 months of gestation – Cattle -3 ½ - 4 months of gestation – Sheep and goat -2 ½-3 months of gestation – Swine -3 months of gestation – Dogs- 5 days after birth ■ Failure of testis to descent into the scrotum is called as cryptorchidism. ■ The retained testis is called as cryptorchid testis. Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

SCROTUM

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Location of scrotum ■ The scrotum is located between the thighs except in boar and tom cat. ■ In boar and tom cat, it is located caudal to the thighs, caudal and ventral to the ischiatic arch. ■ Scrotum is externally composed of skin which is relatively devoid of hair except in ram (male sheep) and buck (male goat) and tom cat (male cat).

Structure of Scrotum  Beneath the scrotal skin is the dartos muscle, which consists of fibroelastic tissue and unstripped cremaster muscle.  Dartos muscle divides scrotum in to two halves.

the

 It is closely attached with the tunica vaginalis and scrotal ligament which is a remnant of gubernaculum.

Scrotal ligament  The Testis and epididymis are fixed in scrotum by means of scrotal ligament which is attached near the tail of the epididymis and mesorchium.  The Scrotal ligament is absent in bulls.

The Scrotum has the following 8 layers from externally ■ Epidermis – outermost layer ■ Dermis ■ Dartos muscle ■ Connective tissue

■ Parietal layer of vaginal process (PVP) ■ Cavity of vaginal process (CV)

■ Visceral layer of vaginal process (VVP) ■ Tunica albuginea Dr. GN Purohit - Confidential - Content & Video Not Permitted for Commercial Purposes

Seminiferous Tubules  The Tunica albuginea penetrate the testicular parenchyma to join at mediastinum.  These fibrous septa divide the testicular parenchyma into lobules.  The lobules are having the highly coiled seminiferous tubules.  About 75% of the testicular mass is composed of seminiferous tubules.

Blood supply to scrotum External pudental artery and also Internal pudental artery in boar and cat. Nerve supply to scrotum Genital nerve which is a branch of genito-femoral nerve arising from second to fourth lumbar nerve and perineal nerve. Smooth muscle of the scrotum is supplied by the spermatic plexus from pelvic plexus.

Functions of scortum It encloses the male gonad It provides suspensory attachment of testes with remaining animal body. It protects the testes from adverse stress conditions. It serves as a thermic chest of male gonad. It maintains gonadal temperature through thermo regulatory mechanism for optimum spermatogenesis

■ The Scrotum, cremaster muscle and pampiniform plexus maintain the testicular temperature.

The scrotum becomes flaccid and elongated due to relaxation of the dartos and cremast...