Cloning has advanced through the recent demonstrations that it is feasible to produce, in principle and with significant effort, an unlimited number of individuals of identical genotype from differentiated cell lines that have been frozen and thawed. These advances have been based upon understanding the importance of interactions between the stage of the cell cycle of both the oocyte and donor cell for the success of the nuclear transfer. Whilst the impact of the biological advance is immense for biomedicai applications, the significance is less clear for livestock breeding. In our view the scientific issues for breeding programmes lie in whether clones can increase genetic progress without a cost to biodiversity. Biodiversity within a species may be categorized as: (i) betvjeen-breed variation; (ix) genetic variation among parents within breeds; (iii) genetic variation among individuals within a farm; and (iv) allelic variation within an individual. In the face of a rapid global decline in breed diversity, cloning, in particular cloning of adults, may be an important route to protect biodiversity since it may allow far more genetic variation to be made available for new breed development in the future than is practicable at present. For variation among parents, the judicious use of clones may give significantly faster rates of progress without increasing the rate of loss of genetic variation and furthermore can help improve traits associated with health and welfare which are at present less tractable than, say, milk yield. Local diversity within a farm may be greatly affected if cloning is utilized to disseminate genetic progress widely and more answers are required on the importance of genetic variation within any one locality either in buffering diseases or ameliorating other management problems. Experience from clonal forestry can provide some indications but now there are models capable of answering this question directly in livestock. Allelic variation within individuals per se is not generally advantageous but at loci where it is identified to he beneficial, the use of cloning may exploit it more widely.

This review addresses the reasons for the lack of progress in the control of superovulation and highlights the importance of understanding the mechanisms underlying follicular development. The present inability to provide large numbers of viable embryos from selected females still restricts genetic improvement, whilst variability in ovarian response to hormones limit the present capacity for increasing reproductive efficiency.

Females are born with a large store of eggs which rapidly declines as puberty approaches. If these oocytes are normal then there is scope for increasing the reproductive potential of selected females. Oocytes must reach a certain size before they can complete all stages of development and the final changes that occur late in follicular development. It is likely that oocytes that do not produce specific factors at precise stages of development will not be viable. Hence, it is important to characterize oocyte secreted factors since there are potential indicators of oocyte quality.

The mechanisms that determine ovulation rate have still not been fully elucidated. Indeed follicular atresia, the process whereby follicles regress, is still not known. A better understanding of these processes should prove pivotal for the synchronization of follicular growth, for more precise oestrous synchronization and improved superovulatory response.

Nutrition can influence a whole range of reproductive parameters however, the pathways through which nutrition acts have not been fully elucidated. Metabolic hormones, particularly insulin and IGFs, appear to interact with gonadotrophins at the level of the gonads. Certainly gonadotropins provide the primary drive for the growth of follicles in the later stages of development and both insulin and IGF-1, possibly IGF-2, synergize with gonadotrophins to stimulate cell proliferation and hormone production. More research is required to determine the effects of other growth factors and their interaction with gonadotropins.

There is evidence, particularly from studies with rodents, that steroids can also modulate follicular growth and development, although information is very limited for ruminants. There may be a rôle for oestrogens in synchronizing follicular waves, to aid in oestrous synchronization regimes and for removing the dominant follicle to achieve improved superovulatory responses. However more information is required to determine whether these are feasible approaches.

Heritability for litter size is higher in sheep than in cattle. Exogenous gonadotropins are a commercially ineffective means of inducing twinning in sheep and cattle. Although there are differences in circulating gonadotropin concentrations, the mechanism(s) responsible for the high ovulation appear to reside essentially within the ovaries. The locus of the Booroola gene, a major gene for ovulation rate, has been established but not specifically identified. However sheep possessing major genes do provide extremely valuable models for investigating the mechanisms controlling ovulation rate, including a direct contrast to mono-ovulatory species such as cattle.

In conclusion, the relationship between oocyte quality, in both healthy follicles and those follicles destined for atresia, must be resolved before the future potential for increasing embryo yield can be predicted. In addition, a greater understanding of the factors affecting folliculogenesis in ruminants should ensure that the full benefits ensuing from the precise control of ovarian function are achieved. The improved use of artificial insemination and embryo transfer that would ensue from a greater understanding of the processes of folliculo genesis, coupled with the new technologies of genome and linkage mapping, should ensure a more rapid rate of genetic gain.

Artificial insemination, superovulation and embryo transfer have had beneficial impacts on animal production but a limiting factor to realizing the full potential of these techniques and of other reproductive technologies is the availability of fertile oocytes. To overcome this problem, methods for maturing oocytes in vitro (IVM) have been developed. The production of bovine embryos by IVM is in commercial use but the rate of success and quality of embryos is low. The lack of success may be due to the quality of oocytes that are being matured and it would be preferable to utilize the abundant source of immature oocytes from preantral and primordial follicles by developing systems for in vitro growth (IVG). Several culture systems that utilize early growing follicles as a source of oocytes have been developed for laboratory species and these have been successful in producing live young. IVG in association with IVM and cryopreservation have the potential to maximize the genetic potential of high genetic merit females and shorten generation intervals. This paper presents the current status of technology for the in vitro growth and development of immature oocytes, in vitro maturation and cryopreservation of germ cells in domestic ruminants.

The effects of high and low dietary dietary intake on reproduction in female cattle and sheep will be considered at the level of the pituitary gland, ovary and uterus. In sheep, increased dietary intake for a relatively short time will increase ovulation rate, by increasing gonadotropin secretion. Dietary intake can affect steroids such as progesterone and also intra-follicular concentrations of some growth factors such as IGF-1 and IGF-2. The effects of altered energy intake on gonadotropins and steroids in cattle are not as repeatable as those in sheep but follicular growth rates can be altered. High nutrition has a negative effect on oocyte quality, with animals on ad-libitum high energy diets particularly at risk. Overfeeding can decrease embryo quality in both sheep and cattle and it appears that this results from changes primarily at the level of the follicle or oocyte. Restricted nutrition for a short time will enhance pregnancy rates in cattle; most of this benefit appears to occur if food is restricted before insemination. Thus feeding levels before mating are particularly important to subsequent reproductive success. High dietary crude protein may decrease pregnancy rate in lactating cows. In ewes and heifers supplementation with urea failed to have any effect on pregnancy rates when good quality embryos were transferred to recipient animals exposed to high dietary crude protein. In donor ewes there were adverse effects on early embryo development following urea treatment, suggesting that the mechanism affecting the reproductive process was primarily operating at the level of the oocyte. Collectively, these data identify the overall deleterious effects of high dietary intake and excess crude protein on fertility and highlight the importance of dietary intake before ovulation on the likelihood of establishing a viable pregnancy.

The emphasis in nutritional studies on foetal growth has now moved from the last trimester of pregnancy, when most of the increase in foetal size takes place, to earlier stages of pregnancy that coincide with foetal organogenesis and tissue hyperplasia. At these stages absolute nutrient requirements for foetal growth are small but foetal metabolic activity and specific growth rate are high. It is thus a time when nutrient supply interacts with maternal factors such as size, body condition and degree of maturity to influence placental growth and set the subsequent pattern of nutrient partitioning between the gravid uterus and maternal body.

Throughout pregnancy the maternal diet controls foetal growth both directly, by supplying essential nutrients and indirectly, by altering the expression of the maternal and foetal endocrine mechanisms that regulate the uptake and utilization of these nutrients by the conceptus. Nutritional effects on the endocrine environment of the embryo during the early stages of cell division can alter the subsequent foetal growth trajectory and size at birth; so too can current in vitro systems for oocyte maturation and embryo culture up to the blastocyst stage. There is increasing evidence that subtle alterations in nutrient supply during critical periods of embryonic and foetal life can impart a legacy of growth and developmental changes that affect neonatal survival and adult performance. Identifying the specific nutrients that programme these effects and understanding their mode of action should provide new management strategies for ensuring that nutritional regimens from oocyte to newborn are such that they maximize neonatal viability and enable animals to express their true genetic potential for production.

Fertility is an important component of herd production efficiency, as each additional oestrous cycle that does not result in a planned pregnancy adds to the cost of dairy farming. In addition to the negative impact on milk production, the high costs of veterinary intervention, re-insemination and herd replacement, subfertility can affect the rate of genetic gain in traits of economic merit. In contrast with the steady increase in average milk yield per cow during the last 30 years, there has been a decline in conception rate to artificial insemination in both the USA and in the UK.

The genetic correlation between yield and fertility has been equivocal. Attempts to improve the reproductive efficiency in dairy cattle through breeding and selection have been frustrated to date by the lack of heritable reproductive parameters conducive to high fertility. However, the traditional fertility parameters of interval to first service, services per conception, days open and calving intervals are highly influenced by managerial decisions and have, as expected, heritabilities too low to permit a meaningful genetic gain through selection. An alternative approach is to use the growing body of evidence that the majority of endocrine factors affecting reproduction are a result of gene expression at the hypothalamic, pituitary, ovarian or uterine level. Mechanisms such as commencement of post-partum cyclicity, follicle wave patterns, manifestation of oestrus, luteal competence and level of embryo mortality are appropriate for study. Research is required to investigate the genetic component of the variation between animals in these parameters, their phenotypic and genetic correlations with fertility and their association with other production traits.

In summary: (a) subfertility is a syndrome with multiple causes and only the symptom in common; (b) improvement in fertility will continue to be frustrated until recorded traits provide more accurate estimates of breeding values; (c) techniques are now available to estimate the genetic variation in physiological components conducive to improved reproductive efficiency; (d) once the heritable components of fertility are identified, these tools could be introduced into progeny testing and breeding nuclei, from which the genetic improvement can be widely disseminated. Selection for those components with sufficient genetic variation will result in the improvement of the integral endocrine and other physiological mechanisms favourably correlated with high fertility (e) these tools may also assist in detecting quantitative trait loci for faster genetic gain through markers-assisted selection.