Biotech in Animal Reproduction

Terms Artificial Insemination Sperm Semen The deposition of spermatozoa in the female by artificial means Sperm The male sex cell, produced by the testes Semen A fluid substance produced by the male reproductive system containing spermatozoa suspended in secretions of the accessory glands

Terms Sire Quarantine Protectant The male parent Process that requires animals to be kept in isolation for a period of time to make sure they do not have a disease Protectant Substance such as glycerin added to semen before it is frozen

Terms Estrus Artificial Vagina Ejaculation The period that a female animal is ready for mating Artificial Vagina A rigid tube used to collect semen for artificial insemination Ejaculation The discharge of semen from the reproductive tract of the male

Terms Conception Motility Extender The beginning of gestation; when a sperm fertilized an ovum Motility Active movement in artificial insemination of the sperm Extender A substance, such a milk, egg yolk, glycerin, or/or antibiotics, that is added to semen to dilute and protect it

Terms Straw Hormone Estrus Cycle Small hollow tubes that contain frozen semen Hormone A chemical substance produced by the body or introduced into the body to produce a specific effect Estrus Cycle The reproductive cycle in nonprimates

Terms Follicle Stimulating Hormone (FSH) Follicle Embryo Transfer A hormone to induce estrus Follicle A small blisterlike development on the surface of the ovary that contains the developing ovum Embryo Transfer The process of removing an embryo from a superior female and implanting it into an inferior female.

Terms Dam Progeny Testing Donor Cow The female parent Test of the value of an individual's genotype by looking at the progeny produced by different matings Donor Cow A cow that is of unusual value as a breeding animal, used for embryos

Terms Recipient Cow Superovulation Prostaglandins A cow of ordinary value that will be implanted with a superior female’s embryo Superovulation Process of injecting the donor animal with a follicle stimulating hormone which will cause the animal to release several eggs instead of just one Prostaglandins Hormones that will make a recipient cow come into estrus

Terms Corpus Luteum Sperm Sexing Cytometer Cell Sorter Active tissue that produces progesterone if conception occurs (Yellow Body) Sperm Sexing Separating the female (X) cell from the male (Y) cell in order to produce only male or only female offspring Cytometer Cell Sorter An instrument used to sort (X) sperm from (Y) sperm according to the amount of light given off by the chromosome

In this chapter: Biotech in animal breeding Advantages of artificial insemination Process of artificial insemination Semen processing, storage and shipment Estrus synchronization Egg production Advantages of embryo transfer Process of embryo transfer Advantages of sexed semen Sperm sorting An Overview Every aspect of the livestock industry is dependent on animal reproduction!!!!

Artificial Insemination The deposition of spermatozoa in the female by artificial means Sperm The male sex cell, produced by the testicles Semen A fluid substance produced by the male reproductive system containing spermatozoa suspended in secretions of the accessory glands

Advantages Use of the highest quality animal available Cost of semen vs whole animal Availability of records # of offspring (potential for 40,000+ from one male) Goals & selectability for herd needs Farm safety (No bull/aggressive male) Reduced transmission of disease (genetic & sexual) No quarantine time on out of country stock Reduced sire cost to producers Increased uniformity

Sire Progeny Testing The male parent Test of the value of an individual's genotype by looking at the progeny produced by different matings

Development As far back as the middle ages Collection of prized enemy stallions First successful insemination: 1780 Lazarro Spallanzani Dog Little economic value at the time 1st large scale use Russians 1900 Horse repopulation Cattle & Sheep 1920/30s

How it works:

Problems? Preserving live sperm Freezing! ALL HAIL THE 1950s!! 2-3 day life span Freezing! Relatively few sperm survived ALL HAIL THE 1950s!! Protectant: Substance such as glycerin added to semen before it is frozen Temperature lowered at a specific rate until -320°F Will remain viable for years (30+!)

Any drawbacks? Labor intensive! Subject to potential abuse? Monitor for estrus Singular restraint & insemination Requires skilled technician Subject to potential abuse? Accentuate damage of poor sire? Additional capital & management Others?

Semen Collection & Processing Dummy animal & artificial vagina Sperm examination Number (15,000,000+ = normal) Motility Shape Add extenders Semen Collection & Processing Artificial Vagina A rigid tube used to collect semen for artificial insemination Extender A substance, such a milk, egg yolk, glycerin, and/or antibiotics, that is added to semen to dilute, nourish and protect it

Volume of Semen and Conc. Per Insemination Species Dilution Ratio Volume/ Service # Sperm/ Insemination Bull 1:200 .5 cc 15,000,000 Stallion ---- 5-10 cc 250,000,000 Boar 1:8 50-100 cc 2,000,000 Ram 1:1 50,000,000 Volume of Semen and Conc. Per Insemination *1 cc is the volume of semen contained in each straw of bull and ram semen

Semen Collection by Specie http://www.youtube.com/watch?v=y6IEk9RNvA4 http://www.youtube.com/watch?v=lzJhhfyoxY0 http://www.youtube.com/watch?v=qgf6h588v-I Semen Collection by Specie Bull Boar Turkey

Straws: Small hollow tubes that contain frozen semen Examined and packaged in straws Stored at -320°F in liquid nitrogen Thawed immediately before placed in female Proper temp & speed Straws: Small hollow tubes that contain frozen semen

When to breed:

Heat Detection Visible activity: But what if you’re not there? Excitability, restlessness & noisy Attempt to ride other cows Stand to be ridden by other cows But what if you’re not there? Ruffled hair on tale head High tail head appearance Genital mucus Swollen vulva Tail chalk, kamars, heat detect systems *In Cattle

Repro Cycle in Farm Animals Species Length of Estrus Cycle (d) Length of Estrus Time of Ovulation Gestation Length (d) Age @ Puberty (m) Avg. Range Avg Mare 21 10-37 5-6 d 1-14 d 24-28 h before end of estrus 336 310-350 10-12 Cow 19-21 16-24 16-20 h 8-30h 10-14h after end of estrus 281 274-291 4-8 Ewe 16 14-20 30 h 20-42h 1 h before end of estrus 150 140-160 Sow 18-24 1-5d 18-60h after estrus begins 112 111-115 5-7

Factors for Consideration Highest conception rates = 60-90 days after calving Breed from 9 hrs following heat to 24 hrs after heat Sperm will live about 24 hrs in the female repro tract True heat is when a cow will stand to be ridden Fertilization will occur immediately if sperm is present when the egg is released

Think about It: Discuss how AI might be of economic importance to a livestock producer. What is the female hormone produced by the ovary that regulates the heat period? What is parturition? What is semen extender?

Control of the Estrus Cycle Elimination of consistent monitoring Big savings! Time Labor Costs Hormone regulation!! Control of the Estrus Cycle Estrus Cycle The reproductive cycle in non-primates

Hormones!! FSH LH Progesterone Prostaglandin Estrogen A hormone to induce estrus & stimulate ovum release LH Triggers ovulation Progesterone Produced by CL to maintain pregnancy Prostaglandin Hormones that will make a recipient cow come into estrus Estrogen Released by the follicle; signals heat

In other words: Organ/Site Hormone Released Target Site Result Pituitary Gland FSH Ovary Egg Mature/Release Follicle Estrogen Heat LH Ovulation Corpus Luteum Progesterone Uterus Maintains Pregnancy Prostaglandin If not pregnant Blood Balanced Cycle Over

FSH Stimulates follicle Begin cycle at injection Synch groups of animals to ovulate at the same time SAVES BIG! Resources at breeding Resources at calving/farrowing Groups of same age animals

Embryo Transfer http://www.youtube.com/watch?v=hRUJSMfFgfc

Benefits of Embryo Transfer Advancement of genetics from the dam Progeny testing of females Import and export without quarantine Dual production system (milk & meat) Twinning Conversion from grade to registered/purebred

Disadvantages Narrowed genetic base Loss of diversity Case and point: Guernsey cattle

Process of Embryo Transfer Selection of donor and recipient cows Donor depends on: Milking ability Ability to grow Reproductive capacity Show appeal Recipient depends on: Health Ability to maintain pregnancy Ability to deliver a healthy calf Process of Embryo Transfer Donor A cow that is of unusual value as a breeding animal, used for embryos Recipient A cow of ordinary value that will be implanted with a superior female’s embryo

Get them In Synch!

Not like that! Prostaglandin Monitor CL Implant embryos Superovulation Donor Animals Recipient Animals Superovulation 12-15 eggs Inject FSH Several follicles to shelter growing eggs Prostaglandin Causes estrus Insemination Collection Grown for ~1 week Prostaglandin Causes estrus Monitor CL Implant embryos In uterus

The Flush! And not like this either!!! Or this!!

The REAL Flush! Long rubber catheter Inflatable bulb Passed through cervix to uterine horn Inflatable bulb Seals entrance to the uterus Solution injection Fill fallopian tubes and uterus Drainage Solution and eggs drain into collection cylinder ~6 embryos per flush And afterwards? The uterus is flushed again with a solution that will kill any remaining embryos to prevent infection.

The numbers Up to 30 embryos could be obtained in one flush (usually 6-7) Survival to term: 50-70% Superovulated vs non Non= 5 calves/year Super= 9-12 calves/year Recovery = 50-90%* Cattle = 50-80% Horses = 40-90% *Nonsurgical methods

Think About It What support technology & procedures are involved in ET? What are the advantages to embryo transfer? What are the expected result rates of ET? What is superovulation and why is it used? Indicate expected recovery rates concerned with ET? What is estrus synch?

Sperm Sexing Why? Depends on industry Dairy: Females = milk producers Swine: Boar meat = odorous & unacceptable Beef: Males = bull studs for breeding Poultry: Females = layers

Chromosomes A refresher: Females have 2 X chromosomes Males have an X and a Y When eggs are formed they all have an X Sex determined by which sperm fertilized the egg

Fluorescent dye adheres to sperm DNA 1989: Method patented Fluorescent dye adheres to sperm DNA Female chromosome has more DNA More dye in the female chromosome (by 2.8-7.5%)!! Sperm Sexing Sperm Sexing: Separating the female (X) cell from the male (Y) cell in order to produce only male or only female offspring Cytometer Cell Sorter: An instrument used to sort (X) sperm from (Y) sperm according to the amount of light given off by the chromosome

And now? And now? Guarantee 85-95% accuracy (cattle)