Artificial insemination (AI) in alpacas provides several advantages but also faces challenges. AI allows for the widespread use of elite males to improve herd genetics. It facilitates disease prevention but requires reliable semen collection techniques as alpaca semen has low volume, concentration, and motility. Current limitations include a lack of methods for chilled or frozen sperm storage. Successful AI also depends on inducing ovulation at the right time.

1. Artificial insemination in alpacas Jorge Reyna Alpaca Breeding Technologies

2. Why alpacas are becoming so popular worldwide? Fibre production Ability to tolerate harsh climatic conditions Less erosive effect (fibro elastic pads) Meat production Skin Fuel Transport Companion animals

3. Genetic improvement data collection statistical and prediction models selection reproductive technologies (AI, ET, IVP) as a tool to disseminate the best genes in the population.

4. Why the genetic improvement is slow in alpacas? Males reach puberty from 1-3 years Long gestation length (11.5 months) Limited number of offspring in females in the whole reproductive life (3-4 in Peru).

5. Advantages of AI in alpacas I Widespread use of elite sires Facilitates progeny testing Improved performance of the national herd Permits crossbreeding to change a production trait Accelerates introduction of new genetics

6. Advantages of AI in alpacas II Reduces risk of spreading sexually transmitted and other (lice, JD) diseases Eliminates the need of transport of animals, reducing the need for “mobile matings” Provides a useful tool for investigating reproductive physiology

7. Current limitations of the AI in alpacas Need reliable technique to collect sperm. Length of copulation 25.65 minutes (17 to 43) The unique mucoid character of the sperm. Low spermatozoa concentration and low motility in the ejaculate. Frequency of use of the male. Lack of techniques to store sperm in chilled or frozen form. The characteristic of induced ovulation in the female – timing of AI?

8. Ejaculatory process in alpacas Copula takes up to 40 minutes. Urethral contractions are distributed evenly during the ejaculatory process. Ejaculation process is constantly (dribbling) Seminal plasma is gelatinous and holds sperm until ovulation takes place. > time of copulation > chance to find sperm on the ejaculate.

9. Semen collection techniques in alpacas Intravaginal condom (1952) Electroejaculation (1968) Intravaginal sponge (1970) Artificial vagina (1983) Vaginal aspiration (1994) Artificial reproductive tract (2005)

10. Collection by Artificial Reproductive Tract (ART) 1. Prep. artificial vaginal 2. Fit into a mannequin 3. Mating process 4.Collection of ejaculate 5 . Assessments

11. Parameters of alpaca semen (5 males, 3 locations, 15 collections) Macroscopic evaluation: Volume 2.57 ml (1 to 6) Colour (Milky white) Consistency (Viscous, gelatinous) Microscopic evaluation: Motility 56.67 % (10 to 70%) Concentration 87.43 x 10 6 sperm/ml (4.8 - 220) Live sperm (58 – 83 %) Abnormalities (heads 3-13%, tails 9-15%)

12. Alpaca sperm appearance White creamy appearance with foam on top

13. Alpaca sperm viscosity Alpaca sperm is highly viscous, which makes it difficult to obtain a homogenous sample when it is mixed with extender for artificial insemination and/or freezing procedures

14. Alpaca sperm under the microscope

15. Concentration of sperm from farm animals (million/ml) Rooster 3000 - 7000 Ram 2000 - 3000 Bull 800 - 2000 Boar 200 - 300 Stallion 150 - 300 Cat 168 - 361 Rabbit 150 - 500 Alpaca 48 - 124

16. Sperm processing Extenders are required to protect semen against cold shock during freezing procedures. The first step is liquefaction of the semen to mix the extender with the sperm Incubation at 37C for 8 h. Hydrolytic enzymes (Trypsin, Collagenase, Fibrinolysin, Hyaluronidase ) mechanical stirring other?

17. Cryopreservation An excellent extender has the following functions: Provides nutrients as a source of energy. Protects against the harmful effect of rapid cooling Ensures the pH will be maintained (buffer) Maintains osmotic pressure and electrolyte balance Inhibits bacterial growth Increases the volume of semen Protects sperm cells during freezing

18. Ovulation in alpacas

19. Lifespan of the dominant follicle

20. Preparation of the females for AI Induction of ovulation is required using a vasectomised males or with hormones like GnRH or hCG and inseminate at ????? hours later. Another technique that is being used in Peru is seminal plasma (0.8 -1.0 ml) i.m achieving 60 % of pregnancies.

21. Artificial Insemination Intrauterine insemination: Cervix is stabilised per rectum with the left hand and pipette is introduced and semen deposited in both uterine horns. Laparoscopic insemination: abdominal incisions are required and sedation. Semen is deposited in the uterine horn ipsilateral to the ovary containing an ovulatory-size follicle.

22. Contact us Jorge Reyna BSc (Hons), MScVetSc (Sydney Univ.) 2/122 Newington Rd, Petersham, NSW 2049 Australia Phone: +61 2 9568 1370 Mobile: +61 428 ALPACA E-mail: [email_address]