Semen collection in the ostrich

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1 Semen collection in the ostrich Irek A. Malecki 1, Paulina K. Rybnik 2, Jaroslaw O. Horbanczuk 2, Ewa Lukaszewicz 3 and Henryk Naranowicz 4 1 School of Animal Biology M085, Faculty of Natural and Agricultural Sciences, University of Western Australia, Crawley WA 6009, Australia 2 Institute of Genetics and Animal Breeding, Polish Academy of Sciences, Department of Animal Sciences, Jastrzebiec, Wolka Kossowska, Poland 3 Department of Poultry Breeding, Wroclaw University of Environmental and Life Sciences, Wroclaw, Poland 4 Stypulow Ostriches, Kozuchow n/zielona Gora, Poland Introduction The ostrich industry originated in Africa and has later spread to almost all parts of the world. While the history of ostrich farming is more than 200 years old a method for obtaining semen from ostriches was only published 30 years ago. Von Rautenfeld (1977) described the manual massage method for collecting semen from ostriches that was further improved and applied by Bertshinger et al. (1992), Irons et al. (1996) and Hemberger et al. (2001). The manual massage method, however, showed limited success and has been problematical due to stress to the birds and the handlers during restraining and phallus manipulation. The manual massage method was not reliable, could not guarantee quality ejaculates routinely and therefore ostrich farming did not adopt this method. In practice, semen is collected for evaluation of male fertility, diagnosis of fertility problems, semen preservation and for artificial insemination. For such purpose semen needs to be physiologically normal and attempts to develop a stress free method that would evoke normal ejaculation were subsequently made (Rozenboim et al., 1996, 1999, 2003; Ya-jie et al., 2001). In those attempts docile ostrich pairs that cooperated with the handlers were used and ejaculates were collected by interrupting mating with the artificial cloaca (AC) (Malecki et al., 1997a,b), although Rozenboim et al. (2003) found that the AC was stressful for their pairs and a vacuum system used for collecting semen from turkeys was giving better results. Despite those attempts the training method and the collection techniques have not been clearly established. Moreover, these methods require co-operation of a pair of birds that at times can be problematical and therefore do not often guarantee ejaculates on demand. The method not involving a female, the nonteaser approach, was first developed in ratites for the emu (Malecki et al., 1997a). The method is based on development of desirable human-male bird interactions. In the

2 ostrich, this concept was proven by Malecki and Martin (2005) who showed that those male ostriches that respond to humans by squatting and performing a ground display (kantling) are good candidates to non-teaser training and can be trained to mount the dummy (Malecki and Martin, 2005). Later research has provided more knowledge on training conditions for the teaser and non-teaser methods in ostriches and some quantitative characteristics of the ostrich ejaculate (Rybnik et al. 2007). The methods described in this paper are a summary of our recent work and some former research carried out on this topic by other researchers. Semen collection Manual massage method The male ostrich may be confined in a specially constructed crush in order to prevent injury to the bird and the handlers (Fig. 1) or may just be held in the corner of the yard. Once the bird is restrained, the phallus is extruded out of the cloaca and held down with one hand, using a cloth to allow a firmer grip. The fingers of the right hand are introduced into the phallic groove and passed deeply into the cloaca until the semen papillae are located (Fig. 2). Figure 1. The male ostrich restrained by handlers for semen collection by massage. Figure 2. Massage of the extruded ostrich phallus. Gentle massage of the papillae may lead to ejaculation and then semen would run down the phallic groove. Teaser method The artificial cloaca (AC) is prepared with warm water and taken to the birds (Malecki et al., 1997a, Rybnik et al., 2007). The collector appears near the holding yard

3 and walks towards the teaser to induce crouching. The male ostrich approaches the crouching female and attempts to mount her. Then the collector enters the enclosure with AC in his hand and approaches the birds from behind (Fig. 4). While the male is attempting to penetrate the female s cloaca with his partially extruded phallus, the AC is placed next to the female s vent and the phallus is directed into the AC (Fig.5). Once intromission is achieved in the AC the phallus becomes erect and the male ejaculates while arching his neck and swinging his head sideways and grunting. Figure 3. Approach of copulating birds from behind to intercept the phallus. Figure 4. Holding of the AC after intercepting the phallus into the AC. Non-teaser method The AC is inserted into the dummy (Malecki and Martin 2005; Rybnik et al., 2007) and the collector approaches the male enclosure. Once the collector opens the gate and puts a dummy on the floor the male comes up with his wings lifted and attempts to mount the dummy (Fig. 6). The collector lowers himself while the male performs a mount putting the right leg on the dummy, spreading and waving the wings and swinging the head from side to side. When the male achieves intromission in the AC he ejaculates while arching his neck forward and swinging his head sideways and grunting, a sequence of behaviours performed during normal copulation or a teaser method collection (Fig. 7). Once ejaculation is complete the male dismounts and leaves the floor. Characteristics of ostrich ejaculates In the ostrich (Table 1), collecting semen during mating (teaser or non-teaser) appears to yield more sperm per ejaculate than when the manual massage is used.

4 Figure 5. Male ostrich about to mount the dummy. Figure 6. Male ostrich sitting on the dummy. Parameters of semen quality such as motility and proportions of live and normal spermatozoa are not yet defined. They vary considerably within and between males and collection methods. In general, in ejaculates collected by manual massage mass motility was highly variable and considerably reduced (Bertshinger et al., 1992: 0-90%; Hemberger et al., 2001: 42-96%). Table 1. Characteristics of the ostrich ejaculates collected by manual massage, teaser and non-teaser method. 1 Method Volume (ml) Concentration (x 109/mL) Manual massage Bertshinger et al., 1992 Hemberger, et al., 2001 Teaser Rosenboim et al., 2003 Ya-jie et al., 2001 Rybnik et al., 2007 Non-teaser Rybnik et al., Source 3 10 /µl; Male age: 2 years and more; Male age: 2 years In the ejaculates collected by the AC spermatozoa show less variable mass motility (Ya-jie et al., 2001: 78-92%; Rybnik et al., 2007: %) and are within normal range for fresh avian semen. The proportions of live and normal spermatozoa tended to be lower for the manual massage (Bertshinger et al., 1992: 72-77%; Hemberger et al., 2001: 74-94%) than for the teaser method (Rozenboim et al., 2003: 93-96%; Yajie et al., 2001: 87-93%; Rybnik et al. unpublished data: 82-98%) or the non-teaser method (Rybnik et al. - unpublished data: 78-96%).

5 Semen yield When semen was collected once, twice and three times per week using the manual massage the total output decreased with increasing frequency (Hemberger et al., 2001). In the emu, collecting semen twice daily yields more semen and spermatozoa than once a day, when the production of sperm is at its peak (Malecki et al., 1997b). Thus, it can be expected that in the ostrich the frequency higher than once a day would yield more semen than collections once or even less than once a day. However, the teaser or non-teaser approach has not yet been used to determine the effect of collection frequency on semen output so it is still unclear how much semen can be obtained daily or weekly, and what rates of extension of semen for artificial insemination will be achievable. Conclusion Development of the animal friendly method for collecting semen from ostriches has advanced considerably in recent years. Normal ejaculates can be collected regularly but the males need to be trained and important human-bird interaction stimulating birds to perform sexual behaviour need to recognized and taken advantage of in training. Semen collected by the teaser or non-teaser method is of good quality and quantity and is suitable for storage. While our results and those of others indicate normal values for ostrich semen, the daily output of semen and spermatozoa are yet to be determined. The ratite production is inefficient because it is constrained by the biology of the species that are not matched by management strategies or practices. Inadequate rates of egg production, fertility, hatchability, too high embryo mortality, poor chick survival or growth rates are some of the problems. The solution would be a structured breeding program that would achieve rapid genetic improvement in reproductive and economic traits. The ostrich industry appears to be in a good position for development of the artificial insemination (AI) technology although adoption of the AI technology in ostrich breeding will mean substantial changes to the current industry structure. Acknowledgment Our work was supported by the Polish Committee for Scientific Research (Grant no PO6Z ) and Rural Industries Research & Development Corporation (Grant no UWA71A/88A). References: Bertschinger, H.J., Burger, W.P., Soley, J.T. and de Lange, J.H. (1992). Semen collection and evaluation of the male ostrich. Proceeding of the Biennial Congress of the

6 South African Veterinary Association, 7-10 September 1992, Grahamstown, South Africa, pp Hemberger, M.Y., Hospes, R. and Bostedt, H. (2001) Semen collection, examination and spermiogram in ostriches. Reproduction in Domestic Animals 36, Irons, P.C., Bertshinger, H.J., Soley, J.T. and Burger, W.P. (1996). Semen collection and evaluation in the ostrich. In Deeming, D.C. (ed.) Improving our Understanding of Ratites in a Farming Environment. Ratite Conference, Oxfordshire, pp Malecki, I.A. and Martin, G.B. (2005). Development of reproductive technology and fertility assessment for the emu and ostrich farming. Rural Industries Research and Development Corporation, Australia (RIRDC Publication No 05/200). Malecki, I.A., Martin, G.B. and Lindsay, D.R. (1997a). Semen Production by the Male Emu (Dromaius novaehollandiae). 1. Methods for semen collection. Poultry Science 76, Malecki, I.A., Martin, G.B. and Lindsay, D.R. (1997b). Semen Production by the Male Emu (Dromaius novaehollandiae). 2. Effect of collection frequency on the production of semen and spermatozoa. Poultry Science 76, Rozenboim, I., Navot, A., Gvaryahu, G., and Degen, A. (1996). Collecting semen ostrich a new and friendly method. Poultry Science Association, 85 th Annual Meeting, July 8-12, 1996, Louisville, Knetucky, Abs. 72. Rozenboim, I., Navot, A., Snapir, N., Gvaryahu, G., Rosenshtrauch, A., and Degen, A. (1999). A method for obtaining semen from the ostrich (Struthio camelus) and its quantitative and qualitative characteristics. In: Proceedings of the International Conference on Bird Reproduction September 1999, Tours, France (Poster). Rozenboim, I., Navot, A., Snapir, N., Rosenshtrauch, A., El Halawani, M.E., Gvaryahu, G., and Degen, A. (2003). Methods for collecting semen from the ostrich (Struthio camelus) and some of its quantitative and qualitative characteristics. British Poultry Science 44: Rybnik, P.K., Horbanczuk, J.O., Naranowicz, H., Lukaszewicz, E. and Malecki, I.A. (2007). Semen collection in the ostriches using a dummy or a teaser female. British Poultry Science (in press). Von Rautenfeld, D.B. (1977). Mitteilungen zur kunstlichen besamung, geschlechts und altersbestimmung beim strau (Struthio camelus australis, Gurney). Der praktische Tierarzt, 5: Ya-jie, J. I., Yan-bo, Y. and Wu-zi, D. (2001). Studies on ostrich semen character and semen storage at low temperature. Journal of Economic Animal 5,

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