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ARTIFICIAL INSEMINATION(For Veterinarians) 3) Introduction and Semen Quality(For Final Year Students) by Prof.Dr

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ARTIFICIAL INSEMINATION(For Veterinarians) 3) Introduction and Semen Quality(For Final Year Students) by Prof.Dr 1) Successful application of AI in cattle By Prof.Dr. Sayed Hattab Prof. of Theriogenology Alex Universityِ 2) ARTIFICIAL INSEMINATION(for veterinarians) 3) Introduction and Semen Quality(for final year students) By Prof.Dr. AHMED MAMDOUH OSMAN Department of Theriogenology Faculty of Veterinary Medicine Assiut University 4) ARTIFICIAL INSEMINATION By Prof.Dr.Afyfi El-Monofy Faculty of Veterinary Medicine Cairo University 1 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) ARTIFICIAL INSEMINATION (BY Prof.Dr.Sayed hattab) Definition Artificial insemination is a process by which sperm are collected from male, processed, stored and artificially introduced into the female reproductive tract for the purpose of conception. History of A.I 1322, Arab Chieftain stole semen from stallion owned by an enemy 1677, Leeuwenhook used microscope to see sperm 1780, Spallanzani reported a successful AI in dog. 1900, Ivanov of Russia Pioneered AI research in livestock, he was the first to successfully inseminate cattle 1936, Denmark was the first to establish an AI cooperative association 1938 USA first AI cooperative association at New Jersey. 1960, 1970, become popular in cattle. 1990, expansion o horse and swine AI. Objectives of A.I Genetic improvement Diseases control mechanism Possible to increase fertility Decrease breeding expense Advantages of A.I Genetic improvement wide spread use and availability of genetically superior sires. One bull can breed 500,000 cows in a life time 2 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) After death , semen can be used coldest frozen semen 40-45 years old Rapid proof of sire (progeny testing Control of venereal diseases Accurate breeding records Economic cost of very good sire reduced because extend semen cost to maintain sire reduced as do not need as many to breed all the females Safety Disadvantages Estrus detection must be good Need well trained inseminators Use of poor male may increase if not tested well Technology to store cooled or frozen difficult to maintain Need to corral and restrain females Process in A.I Centers Semen collection Methods of semen collection: Artificial vagina: Bull, stallion, buck, ram, dog sometimes poor Stimulate natural copulation Temperature at 45C ( worm water) Pressure (air and worm water) Lubrication (harmless to sperm and not excessive amount) Electro-ejaculation: used in bull and rams 3 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) increase Voltage in pulses (1-10v) low voltage --- accessory gland secretion and high voltage --- ejaculation semen volume higher and sperm concentration lower get urine often poorer quality ejaculate Massage method: Rectal massage of vesicular glands and ampullae Process in A.I Centers Precautions During semen collection Precautions must be considered during collection of semen by AV to obtain good quality and quantity semen: Adjust temperature (45c) AV must be clean, dry and sterilized Do not use any detergents or harmful chemicals to sperm Inner sleeve of AV must be free from any leakages Use of adequate amount of harmless lubricants Adjust the pressure inside AV to stimulate thrust Wash and clean perineal and perpetual regions Avoid the collection in dirty place Teasing the bull by false mount and foreplay Protect the semen collector from cold shock by hand or wool cotton piece Process in A.I Centers Semen evaluation 1) Gross examination Volume Color Consistency PH 4 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) 2) Microscopic Examination Assessment of motility Mass motility Individual motility Assessment of sperm morphology Sperm abnormalities life and dead sperm ripe and unripe sperm -pathological cells Assessment of sperm concentration use of hemocytometer Automatic counter 5 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) 6 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) 7 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) Process in A.I Centers 3) Special laboratory tests Measurement of hygienic quality visual examination catalaze test Measurement of metabolic activity Methylene blue reduction test Electrical activity of semen Fructolysis index Resistant tests resistant to sod chloride sol.1% resistant to cold shock viability test Process in A.I Centers Semen dilution Properties of semen extender: a- Isotonic -salts b- Buffered (PH 6.5) phosphate citrate Tris c- Cold shock protection Lecithin and lipoproteins find in egg yolk and milk d- Nutrients egg yolk and milk fructose and glucose e- Antibiotics Gentamycin Tylosin Lincospectin Penicillin + Streptomycin F-protection for freeze and thaw -glycerol g- preserve fertility -Known and unknown factors found by trial and error 8 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) Process in A.I Centers Processing A) semen collection do not let temperature drop below 35c B) Antibiotic treatment 3-5 minutes before dilution C) pre-dilution start at 35c No glycerol yet up to half final volume D) cooling Cool to 5 c Slow cool over 2 to 4 hours E) Final dilution cooled extender + glycerol F) Equilibration - allow glycerol to act -minimum of 4 hours G) Packaging Plastic straws 0.5ml in US 0.25ml in Europe (French straws) Glass ampoules Old method Pellets No package Most success in swine H) Freezing use of liquid nitrogen vapors first then drop into the Nitrogen -196C I) Storage Dry ice and methanol (old) Liquid nitrogen is now used Do not let tanks dry. 9 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) Semen Tank management Avoid excessive movement or abuse of tank Monitor nitrogen levels routinely and keep a record of nitrogen loss Store the semen in an area with good light but out of direct sunlight. Observe tank daily. Once the tank fails, Nitrogen is lost very rapidly. Keep the tank elevated above the concrete floor or other wet and poorly ventilated surfaces 5-store only the amount of semen needed for six months 10 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) Handling semen within the tank In the typical farm semen tank , dangerous temperatures (- 32 to 112 C) in the upper half of the neck of tank Exposure to these temperature can occur when the semen is transferred from tank to tank or when handling semen within the neck while trying to locate a specific unit of semen Thermal injury to sperm is permanent and can not be corrected by returning semen to liquid nitrogen. Semen Handling practices to minimize thermal damage 1-Transfere of semen between tanks must be coordinated and rapid 2-Avoid unnecessary searching and exposure of semen to dangerously high temperature within neck region 3- Use of Tweezer to transfer the straw to the bath. Quickly lower the rack semen and canister into the tank body. 11 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) Thawing of frozen semen Straws are thawed at 35 C for 12 seconds for 0 .5ml straw or 6 seconds for 0.25 ml straw Do not linger with the straw in the neck of LN storage unit or in the air Remove the straw from the thawing bath immediately after the proper thawing time has passed to prevent excessive worming of the semen after removal of straw carefully Dry the straws with a clean tissue Inspect the straws and discard any with cracks or defective seals 12 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) Assessment of quality of frozen semen Motility 20% of progressively motile spermatozoa are the minimal acceptable motility for frozen semen Number of motile spermatozoa per inseminate depend on breed and fertility level of each bull For most bulls each inseminate should contain at least 10 million motile sperms after thawing regardless of how semen is packaged The influence of the method of thawing on post-thaw motility will also be in the number of motile spermatozoa per inseminate. Acrosomal integrity Spermatozoa with deteriorated or damaged, acrosome unlikely to be capable of fertilization Examination of spermatozoa under differential interference phase contrast microscope The sample is incubated at 37 c and aliqutos are removed for evaluation after 0.2,4,8 hours For semen in egg yolk citrate extender, the semen diluted with two volumes of 0.2% glutaraldehyde in phosphate – buffer saline. 13 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) Insemination Use recto-vaginal technique in cow Time of insemination Cow in estrus in the morning must be inseminated in the evening Cow in estrus in evening must be inseminated in the morning Cite of insemination In anterior half of the cervical canal or just anterior to internal os in the body of the uterus 14 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) ARTIFICIAL INSEMINATION BY Prof.Dr.A.M.Osman (for veterinarians) INTRODUCTION History (Arab Mare) Advantages Precautions to avoid disadvantages Bull Selection (Progeny Test) TOOLS FOR ANIMAL IMPROVEMENTS Selection Breeding Management Reproductive Care Artificial Insemination Liquid Frozen Embryo Transfer : Capacitated Sperm Mature Oocyte ARTIFICIAL INSEMINATION IS ONE OF THE TOOL FOR ANIMAL IMPROVEMENTS AND FROZEN SEMEN USING STRAWS PROVED TO BE THE BEST ALL OVER THE WORLD Requirements for Successful AI Programmes I.Personnels: Willing and Enterest Honest ,Sincere,Patient and Clever Disciplinarian and Faithful Hard worker Scintific background on animal Reproduction Believe in his Work 15 copyright@ E V E T C -O59-I0 Prof.Dr.M.El Naggar (PM) II. Selected Male Genotype : Progeny or Sibling Phynotype : Apparent Characters Double Muscles : From Benilux Countries (Holland,Belgium and Luxumburg) Selected Normandi and Buffalo bulls III.
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