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Semen Banking—Is It Now Feasible for Captive Endangered Species?

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Semen Banking—Is It Now Feasible for Captive Endangered Species? Semen banking—is it now feasible for captive endangered species? W. V. Holt and H. D. M. Moore This article proposes that serious consideration should now be given to the establishment of banks of genetic material from rare and endangered species. In the light of advances in the technology associated with semen assessment and preservation, the limitations that have so far discouraged the practice of semen freezing and banking can now be minimized and circumvented. The authors review these developments and suggest that semen preservation should be recon- sidered as a practical means of preserving the genetic diversity currently contained within captive stocks of wild animals. The World Conservation Strategy published by and freezing was not sufficiently advanced to the International Union for Conservation of permit such a venture but considerable progress Nature and Natural Resources (IUCN) in 1980 has been made since then. Given the current identified the preservation of genetic diversity as rate of scientific development and innovation, an important priority, both for conservation and the view expressed by Foose et al. (1985), that the future development of scientific and indust- another two centuries of research will be re- rial innovation. In order to implement this prin- quired before germplasm banking is a realistic ciple we need to make the best use of new possibility, is unduly pessimistic. technological progress where appropriate. The last decade has seen many developments in the International co-operation in the maintenance technology associated with animal reproduction. by zoos of detailed studbooks, such as those for Ironically, much of the stimulus for this work has the Arabian oryx and Przewalski's horse, means arisen through the need for human infertility that the relative contribution of particular foun- treatments and greater agricultural efficiency. It der or parent animals to the current gene pool is appropriate, therefore, to use the fruits of this can be calculated. Certain individuals may con- research in conservation, countering the effects tribute excessively, minimally or not at all; these of world-wide population pressures. effects, which limit the gene pool for subsequent generations, need to be countered by planned The purpose of this review is to outline ways in breeding programmes (for examples, see Ches- which new advances in artificial insemination ser et al, 1980; Foose, 1980). These typically technology and genetics could now be applied involve the expensive and stressful transport of to achieve this end with mammalian species; we animals, but artificial insemination and embryo recognize that conservation of other phyla is transfer technology can be used both to avoid equally important, but these will not be discussed this procedure and to amplify the international here. In particular this article proposes that seri- distribution of valuable genetic resources. Furth- ous consideration should now be given to the ermore, recent developments in molecular biol- idea of setting up an international repository of ogy should facilitate the genetic analysis of popu- frozen semen from endangered species; ten lations and assist with the unequivocal identifica- years ago the technology for semen assessment tion of parent-offspring relationships. 172 Oiyx Vol 22 No 3, July 1988 Downloaded from https://www.cambridge.org/core. IP address: 170.106.40.40, on 25 Sep 2021 at 23:16:31, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0030605300027800 Artificial insemination in endangered Table 1. Mammalian species that have been successfully species bred by artificial insemination Ten years ago artificial insemination and semen Primates Baboon freezing in cattle breeding were already estab- Chimpanzee Pan troglo- Fritz, 1986; lished procedures, and some advocates of their dytes Martin etal., application in conservation were rather naively Gorilla Gorilla g. 1978. gorilla Douglass, optimistic about applying the techniques to other 1981 species. Hence, some zoos started to collect and Carnivores Cheetah Acinoyx jubatus freeze semen from a wide range of species, with- Puma Felis concolor Moore etal., out due regard to the problems of using the Snow Panthera 1981 semen for insemination. These are underlined leopard uncia when it is considered that, after more than 30 Fox Unspecified Aamdal etal., 1 Q7R years of research farmers still do not use frozen Ungulates Addax Addax Densmore ram or boar semen for breeding because the nasoma- etal., 1987 culatus success rate is uneconomically low. Blackbuck Antilope Holt etal., cervicapra 1988 Despite these problems, artificial insemination Speke's Gazella Boever eta/., has been successfully performed in a number of gazelle spekei 1980 Red deer Cervus Krzywinski and species (Table 1), and the secret of success seems elaphus Jaczewski, to lie in the careful monitoring of semen quality 1978 and timing of inseminations. Since the economic Wapiti Cervus Haigheta/., elaphus 1984 aspects of artificial insemination are of minor Others Giant panda Ailuropoda Moore etal, importance when the aim is propagation of a melanoleuca 1984 rare species, the conservationist is not con- strained to use minimal sperm numbers in in- semination doses, and considerable care can for penetrating the tough outer layers that sur- be taken in introducing the spermatozoa into round the ovum, it has always seemed reason- the female reproductive tract. Catheters to de- able to regard sperm motility as a good indicator liver semen samples can be guided through the of fertility. Unfortunately, the commonest cervix, or directly into the uterine horns, under method of rating sperm motility has always been visual control of the operator using fibreoptic to examine a drop of semen through the micro- endoscopes developed for surgical procedures. scope, making some subjective estimate of These procedures lead to improved pregnancy motility based on a simple scoring system. This rates, compensating for the damaging effects of technique is open to many pitfalls, and has pro- freezing upon sperm longevity and vigour. vided little hard evidence for relationships be- tween motility and fertility. Recent research has shown, however, that Advances in semen assessment sperm motility can be measured objectively Faced with a fresh or frozen semen sample, it is using either time-lapse photography or newer necessary to evaluate its fertilization potential in computerized techniques to analyse images. order to avoid spending large sums on preserv- Measurements of sperm swimming speed (Milli- ing semen that stands minimal chance of ever gan et a/., 1980; Aitken et a/., 1982; Holt et a/., achieving a pregnancy. Clearly, attempting to 1985) established that, for human sperm, swim- predict fertility without performing direct trials is ming speed was a very valuable guide to fertility a major problem, but techniques to achieve this in vitro and in vivo. This now forms the basis of end are currently under development. a diagnostic test for fertility, and there is every reason to believe that it could be usefully applied Since sperm movement is one of the most obvi- to semen from the great apes. So far it has not ous characteristics of a semen sample, and is been possible to validate this technique for ungu- essential both for sperm colonization of the cer- lates, although data gathered in the Institute of vix and oviduct prior to fertilization as well as Zoology, Zoological Society of London, show Semen banking for captive endangered species 173 Downloaded from https://www.cambridge.org/core. IP address: 170.106.40.40, on 25 Sep 2021 at 23:16:31, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0030605300027800 that blackbuck, Pere David's deer and oryx sper- suggests that more parameters, such as the pro- matozoa exhibit sensitive alterations of swim- portion of motile sperm and sperm survival time, ming speed in response to different diluting need to be monitored to obtain good corre- media, as well as to cooling and freezing (see lations with fertility; these problems require Figure 1). Published research on bull semen further technical development, but should be (a) Arabian oryx (b) Pere David's deer Fresh sperm diluted in BWW Fresh sperm diluted in BWW 18 14 10 6 2 i_ Stored in egg yolk/citrate for 24 h at 5°C 18 Diluted in egg yolk/citrate (not cooled) 14 14 - 10 10 - 6 2 1 6 _> - 5 2 Stored in egg yolk/citrate for 48 h at 5°C 18 14 10 6 Diluted in egg yolk/citrate (cooled to 5°C) 2 26 22 22 Frozen /thawed 18 18 14 14 10 10 6 6 2 2 15 30 45 60 75 90 105 120 135 15 30 45 60 75 90 105 Swimming speed (Mm/sec) Figure 1. Frequency histograms showing how the swimming speeds of spermatozoa from (a) Arabian oryx and (b) Pere David's deer responded to the various stages of processing involved in semen freezing. Typically, these entail dilution in a cryoprotective medium containing egg yolk and glycerol, slow cooling to approximately 5°C, then freezing in liquid nitrogen vapour. After thawing, Pere David's deer spermatozoa, which showed little change in speed during processing, recovered full motility. In this example, the poor survival of Arabian oryx spermatozoa was indicated by their serious decline in swimming speed during processing. Frozen/thawed sperm were non-motile. (BWW refers to a physiologically balanced salt solution.) 174 Oryx Vol 22 No 3, July 1988 ' Downloaded from https://www.cambridge.org/core. IP address: 170.106.40.40, on 25 Sep 2021 at 23:16:31, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0030605300027800 solved readily using the sophisticated computer that a universally applicable technique may soon techniques available today. be developed.
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