SEMEN COLLECTION TECHNIQUES IN ALPACAS (from www.alpacareproduction.com) Several techniques have been used in the past to collect semen from alpacas. One of the first techniques used was intravaginal condoms (Mogrovejo 1952). The problem with this technique was that it did not consider that the deposit of sperm in alpacas is intrauterine. Copulation of a male with a female that has a condom did not last more than 5 minutes and the ejaculate was incomplete. A second technique used was electroejaculation (Fernandez-Baca 1970), but the problem was that the semen usually was contaminated with urine, which affects motility and can kill sperm, and it was stressful for the animals and could compromise future reproductive life if not conducted carefully. Another technique described was the use of post copula sperm aspiration from the vagina (PCSAV)(Pastor 1993) . This technique was described as simple and economic to use in order to obtain ejaculates from alpacas to study semen physiology, but the problem was that the sample was mixed with vaginal secretions that may alter semen characteristics. Regrettably there are no further reports on the technique. Finally, the latest technique described is the use of a mannequin (dummy female) and an artificial reproductive tract (AART), which has been reported to be effective to collect quality samples of semen (Aller et al. 1997; Chipana 1997; Perez 1997; Bravo 2001; Davalos and Olazabal 2002; Vaughan 2002; Huanca et al. 2004). The problem with the sperm quality obtained by this latest technique is that it varies within and between males, making it hard to obtain a consistently good quality sample needed to test different freezing protocols. There is no explanation for this problem, but it could be related to the design of the AART and the need to improve it to better simulate the anatomy of the alpaca's reproductive tract. On the other hand, it is not only important to select the animals for acceptance of the mannequin, libido and testicular size, it is also necessary to evaluate consistency in semen production. Studies in the area of freezing of sperm need a reliable technique of collection that guarantees the provision of reasonable ejaculates from the experimental process. Field trials demonstrated that the training of the male to mate with the mannequin is a simple procedure. Animals that had not been used for mating service for a while were keen on mannequin mating and produced an ejaculate at the first attempt. In contrast, animals which had been used regularly for natural matings were reluctant, and they needed audiovisual stimuli in the form of a real female being mated by a male near by. It was frequently observed that some males had difficulties finding the entrance of the vagina. Others were uncomfortable when mating the mannequin, manifested by a constant movement and sometimes withdrawal of the penis and standing up for a few seconds. It was noticed that some animals were scared of the mannequin and moved away. Additionally, it was found that the back height of the mannequin was a bit low for some animals, and a piece of wood was used to increase it and give the animal easy access to the AART (Figure 4). Previous experiments conducted in our laboratory point to the need to modify the mannequin, and especially the AART, to make the animal more comfortable with the mating process and to obtain a better quality of semen. The first field trials showed that the mannequin needs to be rebuilt, using a softer material and probably modifying the shape as well. For some animals it takes time to find the entrance of the vagina and an operator is required to lead the penis. Other animals find the vagina, but may feel uncomfortable, withdraw the penis and stretch their legs, and then come back again. It seems to be a matter of training as well, but it is important to have a design that simulates the female in the copulation position (Figure 5). Also, the external insulation seems to be insufficient when collection is performed outdoors, as the AART goes from 45°C at the start of the collection to 37°C at the first 5 minutes, 36.5°C at 10 minutes, 36°C at 15 minutes and 35°C at 20 minutes. The electric blanket that covers the collection tube seems to be effective at maintaining the temperature indoors, keeping the initial temperature of 40°C after 45 minutes, but when it was used outdoors the collecting tube lost 3°C. The new design has been built and is double insulated (internal/external) to avoid the temperature drop (Figure 6). It would be desirable to keep the collection tube at 37°C, as temperatures over or under this will affect the Figure 4: Monty mating Consuelo, a mannequin developed by Dr. Jane Vaughan at Belbourie Alpaca Stud, Wilton Park, NSW. Figure 5: A natural mating, inspiration for the new mannequin that is being testing at our laboratory. Figure 6: PC the new mannequin in the posture adopted by the female in real mating. PC has been made w