Investigation of association between bovine viral diarrhoea virus and cervid herpesvirus type-1, and abortion in New Zealand farmed deer.

This study tested for association between bovine viral diarrhoea virus (BVDv) and cervid herpesvirus type-1 (CvHV-1) exposure and abortion in New Zealand farmed red deer. Rising two-year-old (R2, n = 22,130) and mixed-age (MA, n = 36,223) hinds from 87 and 71 herds, respectively, throughout New Zealand were pregnancy tested using ultrasound early in gestation (Scan-1) and 55-89 days later (Scan-2) to detect mid-term abortion. Sera from aborted and non-aborted hinds at Scan-2 were tested for BVDv and CvHV-1 using virus neutralisation tests. Available uteri from aborted hinds and from hinds not rearing a calf to weaning were tested by PCR for herpesvirus DNA. In herds with aborted hinds, 10.3% of 639 R2 and 17.2% of 302 MA hinds were sero-positive for BVDv and 18.6% of 613 R2 and 68.5% of 232 MA hinds were sero-positive for CvHV-1. There was no association between BVDv sero-status and abortion at animal level (R2 p = 0.36, MA p = 0.76) whereas CvHV-1 sero-positivity was negatively associated with abortion in MA hinds (p = 0.01) but not in R2 hinds (p = 0.36), MA). Eleven of 108 uteri from aborted R2 hinds but no MA hinds were positive for herpesvirus DNA. Vaginal samples from four R2 and one MA aborted hinds tested were negative for herpesvirus DNA. A Cervid Rhadinovirus type-2 (CRhV-2) was identified in seven PCR positive uteri samples. Findings suggest that BVDv and CvHV-1 may not be associated with abortion in R2 hinds, but association needs to be tested further in MA hinds. The role of CRhV-2 requires clarification.

Lack of association between Leptospira spp. serovars Hardjobovis and Pomona and pregnancy and mid-term abortion in New Zealand farmed red deer.

This paper investigates Leptospira borgpeterseni serovar Hardjobovis and L. interrogans serovar Pomona as potential causes of sub-optimum pregnancy rates and mid-term abortion in farmed red deer. Rising two-year-old (R2, n = 22,130) and mixed-age (MA, n = 36,223) hinds from 87 and 71 herds, respectively, throughout New Zealand were ultrasound scanned early in gestation (Scan-1) and a sub-sample re-scanned (Scan-2) 55-89 days later and mid-term daily abortion rate calculated. A sub-sample of sera from pregnant and non-pregnant hinds at both scans, both with (case herds) and without aborted hinds was tested for Leptospira using the microscopic agglutination test with titre cut-point ≥1:48 as positive. At Scan-1, 44.3% of 661 and 4.6% of 647 hinds were sero-positive for Hardjobovis and Pomona, respectively. The geometric mean titre (GMT) for Pomona was higher in pregnant than non-pregnant MA hinds (p = 0.015) but not in R2 hinds. At Scan-2 in case herds, 40.3% of 2242 and 7.1% of 2243 hinds were sero-positive for Hardjobovis and Pomona, respectively. There was no association between Hardjobovis or Pomona serology and non-pregnancy (Scan-1) or mid-term abortion (Scan-2) at animal or herd level. In case herds, GMT was higher in non-aborted than aborted hinds for Hardjobovis (p = 0.018) in MA herds and for Pomona in R2 herds (p = 0.006). No uteri from hinds not pregnant or aborting at either scan, or not rearing a calf to weaning, and fetuses as available, were positive on PCR. Evidence is insufficient to confirm that Leptospira Hardjobovis and Pomona play a significant role in sup-optimum early pregnancy or mid-term abortion rates in deer.

The relationship between body-mass and puberty in young red deer (Cervus elaphus) hinds: evidence of early-life effects on permissive live-weight thresholds.

This study, based on the analysis of growth and reproductive records of 6158 young red deer hinds, tested the hypothesis that the nutritional environment in early life influences the permissive body mass threshold for puberty at around 16 months of age. Live-weight at 3 months (i.e. weaning weight) was a proxy for their nutritional environment between birth and weaning, live-weight at 14 months was the indicator of pre-mating body mass, and pregnancy status at 18-19 months was a proxy for entry into puberty. Data were obtained for two sub-populations of hinds, the commercial stud herds, across four consecutive years. The modelled relationships between pre-mating live-weight and the predicted pregnancy rate demonstrated between-year variation for the logistic curves for the commercial herd for the live-weight range from 50 to 90 kg (P<0.001). For the stud herd, the predicted pregnancy rate at any given live-weight did not vary significantly between years (P>0.05) and the pregnancy rates for live-weights <90 kg were higher than for the commercial herd. The regression of average weaning weight on probability of pregnancy showed significant positive associations (P<0.05) at 60 kg, 70 kg and 80 kg (R(2)=0.513, 0.517 and 0.439, respectively). There were no significant regressions at pre-joining live-weights at 90 kg or above (P>0.05). The study supports the central hypothesis of early-life influences on the permissive body mass threshold for entry into puberty.

Seasonal luteal cyclicity of pubertal and adult red deer (Cervus elaphus).

Reproductive failure of rising-two-year-old (R(2)) hinds and seasonal misalignment between calving and pastoral feed production are two factors limiting reproductive productivity of farmed red deer hinds in New Zealand. This study aimed to better understand processes around female puberty and breeding seasonality by describing the potential breeding season (i.e., oestrous cyclicity) of three red deer genotypes. A total of 27 hinds born in December 2005, representing Eastern European (Cervus elaphus hippelaphus), Western European (C.e. scoticus) and F1 crossbred (C.e. hippelaphus×scoticus) red deer, were blood sampled thrice-weekly for 7-8 months (February-September/October) across two years spanning the potential breeding seasons as R(2)'s in 2007 (i.e., puberty) and as adults in 2008. Plasma progesterone profiles were used to construct breeding cycle histories for each hind. Four R(2) hinds failed to initiate oestrous cycles (i.e., puberty failure). The remaining R(2) hinds, including all F1 hinds, exhibited between two and seven oestrous cycles. F1 hinds were significantly earlier to initiate, and later to terminate, cyclic activity, resulting in a longer mean pubertal breeding season (139 days) than for Eastern (86 days) and Western hinds (86 days). However, the data for R(2) hinds are confounded by live-weight, with the F1 hinds being significantly heavier than other genotypes. There were significant correlations between live-weight and seasonality parameters in 2007. All hinds were cyclic as adults in 2008, exhibiting between four and nine oestrous cycles, and a mean breeding season duration of between 132 (Western) and 137 (F1) days. For adult hinds there were no significant genotype differences in cyclic onset and cessation timing, and no observable relationships between live-weight and any reproductive parameter. However, the mean dates for the onset of the breeding season for all genotypes in 2008 were 2-3 weeks later than normally expected for adult hinds in New Zealand. The reasons for this are unclear but may relate to chronic stress of frequent animal handling. The study has demonstrated that puberty in red deer hinds is associated with a shorter potential breeding season than for adult hinds, and that perturbation of breeding activity appears to be quite common, leading to incidences of puberty failure and possibly other aberrant cyclic events. Live-weight×genotype interactions may influence puberty but do not appear to be strongly expressed in adults. However, the relatively late onset of oestrous cyclicity in the adult hinds may be an artefact of the study that has masked genetic influences on seasonal breeding patterns.

Reproductive cycles of deer.

The cervids are a complex assemblage of taxa showing extreme diversity in morphology, physiology, ecology and geographical distribution. Reproductive strategies adopted by various species are also diverse, and include a range from highly seasonal to completely aseasonal birth patterns. The recent growth in knowledge on cervid reproduction is strongly biased towards the larger-bodied, gregarious mixed grazer-browser species that have adapted well to human management and commercialisation. These species tend to represent 'K-selected' climax species characterised by very productive annual breeding success, singleton births and long breeding life (10+ years). Conversely, we know relatively little about the reproductive patterns of the 'r-selected' smaller-bodied, solitary (and often highly territorial), forest-dwelling browser species, often characterised by great fecundity (twinning) and shorter breeding life (<10 years). This group includes many of the endangered cervid taxa. This review extends earlier reviews to include more recent work on cervid reproductive cycles, particularly in relation to environmental factors influencing gestation length.

Effect of melatonin implants on the incidence and timing of puberty in female red deer (Cervus elaphus).

A study was conducted to test the hypotheses that exogenous melatonin treatment of 11-13 month-old red deer hinds: (1) advances the timing of first ovulation, (2) increases the proportion of individuals attaining puberty at ∼16 months of age, and (3) reduces the live-weight threshold for attainment of first pregnancy. A total of 3901 rising-2-year-old (R2) hinds within two herds (A and B) across two years either received single melatonin implants on two occasions in summer (n=1399) or were untreated controls (n=2502). Hinds were joined with stags from mid January to mid May, and were subjected to real-time rectal ultrasonography in early June to assess pregnancy status (proxy for puberty attainment) and foetal age for conception date assignment. Live-weights were recorded for each hind in January (12 months of age) as a proxy for weight at puberty. Melatonin treatment of hinds was associated with a significant advancement in mean conception dates in both herds in both years (P<0.05), with a cohort difference in mean dates between treated and control hinds ranging from 9 to 17 days. Analysis of the temporal distribution of conception dates for each cohort revealed bi-modal or tri-modal patterns of conception indicative of conceptions to first or subsequent ovulations (oestrous cycles). Across all cohorts, melatonin treatment was associated with higher conception rates to first ovulation (P<0.05) resulting in greater overall synchrony of conceptions. Regression analysis demonstrated a significant negative slope for conception date against live-weight (P<0.001), but there was no evidence that this slope varied with treatment, herd or year (P>0.05); for every 10kg increase in live-weight conception date was advanced by an average of 1.3 days. In Herd A, melatonin treatment was associated with significantly higher pregnancy rates in both years (90.3% vs. 78.0% in Year 1 and 84.4% vs. 57.1% in Year 2; P<0.05). The principle effect of melatonin treatment was to increase the pregnancy rate of hinds of low body-mass. In Year 1, at 60kg live-weight a logit regression model indicated a pregnancy rate of 52% for untreated hinds and 83% for treated hinds. At 105kg the rate for both cohorts was 90%. In Herd B, melatonin treatment was associated with higher conception rates in both years but these differences were not significant following correction for slight differences in mean live-weight (P>0.05). The study has demonstrated that factors influencing puberty attainment in R2 red deer hinds can vary between populations. In Herd A, in which body mass of hinds immediately prior to their first potential breeding season may have been the principle limiting factor, melatonin treatment appears to have instigated the pubertal process in hinds that would otherwise be of insufficient body mass.

Evaluation of management variables to advance conception and calving date of red deer (Cervus elaphus) in New Zealand venison production systems.

The ability to shift the supply of New Zealand chilled venison from farmed yearling red deer stags to obtain premium prices in seasonal European markets necessitates early calving of hinds combined with high growth rates of their calves. Two studies over a three-year period evaluated three management variables that offer potential to advance calving date. Under the conditions of the studies there was no consistent evidence that the management practices of early stag introduction, early weaning and enhanced hind nutrition prior to conception (lactation) and pre-calving (third trimester of pregnancy) advanced conception date and calving date in red deer hinds. However, the nutrition effect was diminished by the difficultly in achieving the dietary contrast necessary for the targeted 5kg differentiation in hind live weight at strategic times of the year. Across all hinds there was a significant pre-mating (mid-March) live weight effect on conception day in the one year in which a 5kg difference between nutritional regimens was achieved, but the driver was live weight and not nutrition. There were significant effects of nutrition on calf growth, with the growth rates of calves weaned in mid-March significantly higher when their dams grazed a high plane of nutrition pre-conception. There were significant and consistent inverse relationships between conception day and calving date that implied variation around gestation length, with early- and late-conceiving hinds exhibiting longer and shorter gestation lengths, respectively. Across all treatments, calving date was predicted to advance by approximately 5 days for every 10-day advance in conception date. However, there was a significant carry-over effect of nutrition pre-conception on calving date, with hinds on a high plane of nutrition pre-conception exhibiting shorter (2-4 days) gestation lengths. There were also indications that hinds may manipulate gestation length in response to live weight gain pre-calving. These findings suggest that fetal growth trajectory may be the principle driver of gestation length and calving date. Although there were no direct effects of hind nutrition pre-mating on conception dates, nutrition remains an important component of the management of hinds and their calves in venison production systems. The outcomes of the 3-year program suggest that there are limited opportunities to manipulate calving date through manipulation of management variables.

The effect of conception date on gestation length of red deer (Cervus elaphus).

Recent studies have demonstrated that gestation length of red deer (Cervus elaphus) is highly variable and influenced by various environmental factors, and this may confer survival advantages for neonates. The current study investigated the relationship between conception date and gestation length to test the hypothesis that within-herd synchrony of red deer births is facilitated by a 'push/pull' control over gestation length, such that hinds conceiving early and late in the breeding season have longer and shorter gestation periods, respectively. In Study 1, data on conception and calving dates were obtained for 393 naturally cycling hinds across two herds. In Study 2, conception and calving dates were obtained from 91 hinds in which oestrus/conception were artificially synchronised across a 4-week range of dates spanning the natural rut. Gestation length for each population was analysed by linear regression, fitting conception day followed by terms for the fixed effect which included hind age (pubertal vs. adult), hind genotype (Cervus elaphus scoticus vs. Cervus elaphus hippelaphus and their crossbreds), calf sex, sire genotype (Study 1 only), birth weight and year. In Study 1, both populations of naturally cycling hinds exhibited highly significant (P<0.001) negative slopes (-0.36, -0.49) for the regression of gestation length against conception date, with indications of a significant hind genotype effect favouring shorter overall gestation lengths for crossbred hinds. Other effects for hind age, calf sex, birth weight, sire genotype and year were not significant. In Study 2, in which conception dates were artificially induced, there was a highly significant negative slope (-0.19), with a notable but non-significant effect of hind age favouring shorter overall gestation length for primiparous (pubertal) hinds (P>0.05). Other effects for hind live weight, calf sex and calf birth weight were not significant. All data sets support the hypothesis, and indicate that for every 10 days difference in conception date there was a change in gestation length of 1.9-4.9 days. This hints at the adaptive importance of optimisation of birth date in wild populations of red deer but the precise physiological mechanisms remain to be resolved. It is postulated that variation in fetal age during the latter stages of pregnancy, when feed quality and voluntary feed intake cycles are in a state of flux, may drive differential growth trajectories for early and late conceived fetuses, leading to nutritional control over fetal maturation and induction of parturition. However, consideration is also given to a putative direct effect of prevailing photoperiod on control of parturition processes in red deer.

The temporal relationship between oocyte maturation and early fertilisation events in relation to the pre-ovulatory LH peak and preimplantation embryo development in red deer (Cervus elaphus).

The temporal relationships among oocyte maturation, gamete transport and fertilisation following the pre-ovulatory luteinsing hormone surge in red deer were established; and secondly, early preimplantation development to the blastocyst stage in relation to the onset of oestrus was determined for red deer. In the first series of observations, oestrus was synchronised in April (N=22), for the fixed time recovery of gametes from 0 to 36 h after the estimated pre-ovulatory LH peak. Matings were observed and the time of the LH peak was determined from the retrospective analysis of blood plasma collected at 3h intervals. Gametes were recovered surgically and the meiotic status of follicular and ovulated oocytes assessed. Spermatozoa were recovered from the oviduct and their motility analysed by videomicroscopy. Nineteen of 22 hinds exhibited a pre-ovulatory LH surge and were observed to mate. Oocyte metaphase I occurred between 11 and 18 h, and metaphase II was completed within the follicle between 20 and 25 h following the pre-ovulatory LH peak. Fertilised ova were recovered from 30 to 36 h in both the ampulla and isthmic portions of the oviduct. Motile spermatozoa were first recovered from the isthmus and the ampulla at 13 and 21 h, respectively, after the LH peak. Hyperactive spermatozoa were observed in both the isthmus and the ampulla flushings but only from the eight hinds that had ovulated. In the second series of observations, 16 mature hinds were synchronised and allocated to groups for embryo collection on days 3, 5 and 7 after oestrus. Eight embryos were recovered; an 8-cell at 90 h, 3 morulae at 137, 138 and 186 h, and 4 blastocysts at 180, 182 and 190 h post-mating. Blastocysts were only recovered from the uterine horns and the mean+/-S.E.M. number of nuclei per blastocyst was 93.5+/-10.0 with a range of 66-114 cells. The results of this study will improve the application of assisted reproductive technologies to red deer as they indicate that oocyte maturation, fertilisation and early embryonic development of the red deer is similar to other domestic ruminants with the exception that the red deer embryo enters the uterus at the blastocyst stage.

Gestation length in red deer: genetically determined or environmentally controlled?

The red deer (Cervus elaphus) of European origin (e.g. subspecies scoticus, hispanicus, hippelaphus) is a medium sized (100-150kg mature hind weight) ruminant that exhibits highly seasonally patterns of autumn conceptions and summer births. Historic data indicate average (+/- s.d.) gestation length of 233-234 (+/- 2-4) days. Recently, however, there has been growing awareness that there is considerably greater variation in gestation length than earlier indicated and that there is a significant element of environmental, and possibly even social, control over the duration of pregnancy in this species. Imposition of variable levels of nutrition over late pregnancy of red deer hinds has been observed to influence fetal growth trajectory and gestation length, with no apparent effect on birth weight. This supports a hypothesis that under conditions of modest feed imbalance, variation in gestation length compensates for variation in fetal growth trajectory to ensure optimisation of birth weight. More recent studies on primiparous (24 month old) red deer hinds have identified surprisingly large variation in gestation length (193-263 days) compared with adult hinds (228-243 days), with earlier conceiving individuals within the primiparous cohort expressing significantly longer gestation than the later conceiving hinds, resulting in a higher level of calving synchrony than expected from known conception dates. This introduces an intriguing hypothesis of social indicative effects on parturition timing to promote within-cohort birth synchrony. Collectively, these data debunk the commonly held notion that gestation length of red deer is genetically fixed within strict limits. A review of the literature points to this as possibly a common phenomenon across a range of non-domesticated ruminant species but this conclusion is not supported by numerous conflicting studies on domestic sheep and cattle.

Reproductive performance of pubertal red deer (Cervus elaphus) hinds: effects of genetic introgression of wapiti subspecies on pregnancy rates at 18 months of age.

Low reproductive productivity of young red deer (Cervus elaphus) hinds on New Zealand deer farms appears to reflect high incidences of puberty failure at 16 months of age. This is despite the general attainment of average liveweights 15-25 kg in excess of the accepted minimum threshold for puberty in subspecies of western European origin (scoticus, elaphus and hippelaphus) that form the basis of the national herd. The present study tests the hypotheses that introgression of the larger North American wapiti subspecies (nelsoni, manitobensis and roosevelti) into breeding herds (1) can be assessed from morphological features of individuals, (2) that there is a relationship between the level of wapiti parentage and non-pregnancy rate at 18 months of age (a proxy for puberty failure) and (3) that minimum liveweight thresholds for puberty increase with increasing levels of wapiti parentage. A total of 4329 18-month-old hinds across four "red" deer farms in southern New Zealand were scanned for pregnancy status. Each hind was assigned a wapiti score (WS) as a subjective assessment of the obviousness of wapiti features. Various body measurements were additionally recorded for each hind. A hair sample was collected for DNA analysis (14 markers) to objectively assign subspecies pedigree (i.e. "Elkmeter") on a subset of 1258 individuals. A total of 506 (11.7%) hinds were not pregnant at 18 months of age with rates varying between 4.1 and 37.3% between farms and years. Mean WS differed significantly between farms and reflected the genetic management policy of each farm. WS was positively correlated to Elkmeter for each farm/year (<0.05) although regression slopes varied significantly. WS was able to be adjusted for these differences to assign a corrected WS (CWS) for all 4329 individuals that estimated the proportion wapiti parentage. Discriminant analysis of morphological variables relative to Elkmeter supported the first hypothesis and showed that shoulder height and body length were good indicators of the degree of wapiti parentage within individuals. This enabled the development of an objective estimate of wapiti parentage (EWP). The actual level of such parentage within herds ranged from <5 to >55%. There was a significant negative association between wapiti parentage and pregnancy, which was strongly influenced by liveweight, supporting the second and third hypotheses. This was manifest as marked displacement of pregnancy probability curves in relation to liveweight between genotype groups, particularly for those groups with >20% wapiti parentage. For example, predicted threshold liveweights required to achieve a 90% pregnancy rate for EWP values that represent 0, 10, 20, 30, 40 and 50% wapiti parentage were 81, 81, 85, 106, 127 and approximately 137 kg, respectively. Within the study herds, the majority of hinds of 0-20% wapiti parentage exceeded the predicted 90% threshold liveweights for their genotype cohort. However, hinds with higher levels of wapiti parentage generally fell below the predicted threshold for their genotype group. The data strongly suggest that under liveweight performance levels measured for red deer, hinds with >20% wapiti parentage are at high risk of puberty failure.

Influence of level of nutrition during late pregnancy on reproductive productivity of red deer I. Adult and primiparous hinds gestating red deer calves.

The present study aimed to relate feed intake of red deer hinds during late pregnancy to dam body condition, foetal development and calf growth. Across 3 years, multiparous (n=33) or primiparous (n=18) hinds with known conception dates were housed in individual pens from days 150-220 of pregnancy, during which time they were each offered one of three daily allowances of pelletised rations (11 MJME/kg DM; 16% CP): high (H; ad libitum), medium (M; approximately 30% less; multiparous hinds only) and low (L; approximately 50% less). Restricted intake levels were retrospectively calculated from the mean intake of H hinds in the previous week. Hinds were returned to pasture at day 220 and calving was closely monitored. Liveweights, body condition score (BCS), and lactation score (LS) of hinds were recorded weekly from day 130 of pregnancy until calves were weaned at 12 weeks of age. Calves were tagged and weighed at birth, and subsequently weighed at 7 and 12 weeks of age. Additionally, hinds in the first year of study underwent CT scans on days 150 and 215 of pregnancy to assess compositional changes of the dam and conceptus. Mean daily ad libitum intakes of H hinds increased from 1.8 to 2.0 kg DM (0.6-0.7 MJME/kg0.75) at around day 150 to 2.8-3.2 kg DM (0.8-0.9 MJME/kg0.75) by day 220. Those of M and L hinds peaked at 1.8 and 1.6 kg DM, respectively, by day 220. This was reflected in significant treatment effects on liveweight gain and change in BCS and LS by the time of calving. CT scans indicated a significant treatment effect by day 215 on the mass of hind tissues (fat and lean) and a non-significant trend on conceptus/foetal weights. Despite apparent differences in foetal growth trajectories, there were no discernable treatment effects on sex-adjusted birth weights. However, there was an unexpectedly wide spread in calving dates that reflected considerable variation in gestation length. Furthermore, gestation length was negatively correlated with change in hind liveweight (but not BCS) between days 150 and 220 of pregnancy for all groups of hinds (P<0.05). Of three neonatal calf mortalities, none were related to overweight (dystocia) or underweight (non-viability) calves. Subsequent growth rates (g/day) of surviving calves did not reflect prior treatment of their dams, although variation in birth date influenced weights on specific dates. It is concluded from this study that while variation in nutrition to hinds during the last trimester may strongly influence foetal development, under conditions of modest feed imbalance, variation in gestation length compensates to ensure optimisation of birth weight.

Influence of level of nutrition during late pregnancy on reproductive productivity of red deer (2) Adult hinds gestating wapitixred deer crossbred calves.

The present study aimed to relate feed intake of red deer hinds in the later stages of gestating wapitixred deer crossbred foetuses on dam body condition, gestation length, birth weight and calf growth. Multiparous hinds (N=18) conceiving at known dates to either wapiti (n=12) or red deer (n=6) sires were housed in individual pens from days 150-220 of pregnancy, during which time they were offered either ad libitum access to pelletised rations (n=6 crossbred-bearing hinds [HH] and n=6 red deer-bearing hinds [RH]) or a restricted offer (n=6 crossbred-bearing hinds [HL]) set at 70% of the average ad libitum intake of HH hind in the previous week. Hinds were returned to pasture at day 220 and calving was closely monitored. Liveweights, body condition score (BCS), and lactation score (LS) of hinds were recorded weekly from day 130 of pregnancy until calves were weaned at 12 weeks of age. Calves were tagged and weighed at birth, and subsequently weighed at 7 and 12 weeks of age. HH and RH hinds exhibited similar patterns and levels of MEI/kg0.75, which peaked at 7.8 MJME/kg0.75 at day 220. HL hinds peaked at approximately 5 MJME/kg0.75 and showed significantly lower rates of liveweight gain during pregnancy. Interestingly, both crossbred-bearing groups initiated mammary development in advance of the RH hinds. While there were significant effects of foetal genotype on mean gestation length (239 days versus 234 days for crossbred versus red deer) and mean birth weight (14.5 kg versus 10 kg), the nutritional contrast for gestation length of crossbred-bearing hinds (i.e. HH versus HL) was not significant but approached significance for birth weight (14.5 kg versus 11.9 kg; P=0.06). Regression analysis revealed weak relationships between changes in hind liveweight and gestation length (P>0.05) but a significant relationship with birth weight (P<0.05). However, change in hind BCS was significantly related to both gestation length and birth weight. Crossbred calves reared by HH hinds were 30% heavier at 7 and 12 weeks of age than the red deer calves. However, those reared by HL hinds were significantly lighter than their genotype contemporaries and only marginally heavier than the red deer calves. These results generally contrast with the previous studies on red deer hinds gestating red deer foetuses [Asher, G.W., Mulley, R.C., O'Neill, K.T., Scott, I.C., Jopson, N.B., Littlejohn, R. 2004. Influence of level of nutrition during late pregnancy on reproductive productivity of red deer, (1) Adult and primiparous hinds gestating red deer calves. Anim. Reprod. Sci., in press] and indicate that the genetically determined higher growth requirements of crossbred foetuses may override any mechanism of compensatory control of gestation length at the expense of calf birth weight. Furthermore, there were marked carryover effects of late gestational feeding on crossbred calf growth and their dam's BCS that highlight the high nutritional demands of lactation.

New developments reproductive technologies in deer.

In vitro embryo production is the platform for advanced reproductive technologies, such as cloning. The in vitro embryo production system developed for farmed red deer (Cervus elaphus) evolved along similar lines to that pioneered by other domestic species researchers. However, applying existing in vitro embryo production methods from these other species resulted in limited success and has necessitated developing a species-specific methodology for red deer based on the their physiology. Analysis of oviduct fluid led to the development of a semi-defined fertilization and culture media system, Deer Synthetic Oviduct Fluid (DSOF), which resulted in successful culture of red deer embryos to the blastocyst stage. Transvaginal ultrasound-guided ovarian examination and ovum pickup has enabled the study of seasonality constraint and propagation from selected female genetics, respectively. During the 4-month breeding season (April-July), 15% of cleaved oocytes developed to blastocysts, whereas no blastocysts developed from oocytes collected after July. The process of developing an in vitro embryo production system for farmed red deer may serve as a beneficial model for the propagation of endangered cervine species.

Development of in vitro embryo production systems for red deer (Cervus elaphus). Part 1. Effect of epithelial oviductal monolayers and heparin on in vitro sperm motility and penetration of in vitro matured oocytes.

In vitro fertilisation (IVF) protocols for red deer have yielded low fertilisation rates, with no embryo development beyond the eight-cell stage when heparin was used as the in vitro capacitation agent. As this low fertilisation rate may result from reduced motility, the present study investigated the use of red deer oviduct epithelial cell monolayers (COEM) and conditioned medium (Cm) from the monolayers to maintain red deer sperm motility in vitro. A second experiment compared the fertilisability of red deer sperm pre-incubated for 4-12h on COEM or for 4h in TALP medium supplemented with 20 microg of heparin.COEM was superior in maintaining red deer sperm motility compared with either Sp-TALP alone or Cm (P<0.05). COEM sustained sperm motility at levels comparable to the initial motility over the 24h period. The motility of sperm incubated in Sp-TALP and Cm was similar and had declined to less than 10% by 4h and no motile sperm were observed by 8h. Overall, the penetration rates of in vitro red deer oocytes were low (5-28%) regardless of sperm treatment. Sperm pre-incubated on COEM penetrated more oocytes than sperm incubated with heparin (P<0.001). Penetration rates were similar for 4-12h pre-incubation of sperm on COEM (P>0.50). Penetration rates were greater across all treatments when both sperm and oocytes were co-incubated for 24h compared to 12h (P<0.001). There were no differences in penetration rates among the four donor stags used in the study. It was concluded that COEM sustains red deer sperm motility in vitro during the 24h observation period. Pre-incubating sperm on COEM does increase sperm penetration rates compared with heparin alone, but at a rate too low and variable to be used on a routine basis. Overall, the penetration rates were comparable to those previously reported for red deer even though differences in heparin concentration, fertilisation systems and stags were used.

Development of in vitro embryo production systems for red deer (Cervus elaphus). Part 2. The timing of in vitro nuclear oocyte maturation.

The time course of in vitro red deer nuclear oocyte maturation was determined. Ovaries were obtained at slaughter and oocytes were aspirated from follicles greater than 2mm in diameter. Oocytes with compact cumulus cells were matured in 50 microl microdrops (10 per drop) under mineral oil containing TCM 199 supplemented with 0.33 mM pyruvate, 10 microg LH and FSH, 1 microg oestradiol and 10% foetal bovine serum. Oocytes were matured at 39 degrees C and 5% CO(2) in air. At 3h intervals (0-27 h) oocytes were removed from incubation, cumulus expansion scored and removed, and fixed oocytes in ethanol:acetic acid (3:1) for 48 h. Oocytes were stained with lacmoid (1%) and nuclear maturation assessed. Oocytes were arrested in the germinal vesicle (GV) stage at aspiration and up to 6h of incubation. The nuclear membrane began to disperse after 6h and by 10.6+/-0.6h of incubation 75% of the oocytes exhibited germinal vesicle breakdown (GVBD). The mean time for 50% of the oocytes to reach metaphase one (MI) and metaphase two (MII) was 11.7+/-0.4 and 24.8+/-0.9h, respectively. Cumulus oophorus were tightly compacted at aspiration and did not begin expansion until 12h of culture. Full expansion was complete by 18 h of culture. Corona radiata cells did not begin expansion until 15 h and were fully expanded by 24h. Results indicate that in vitro red deer oocyte maturation follows a similar time course of nuclear maturation as reported for bovine and ovine oocytes.

Development of in vitro embryo production systems for red deer (Cervus elaphus). Part 3. In vitro fertilisation using sheep serum as a capacitating agent and the subsequent birth of calves.

The following experiments investigated the use of sheep serum (SS) as a capacitating agent for red deer (Cervus elaphus) sperm during in vitro fertilisation. Red deer oocytes were collected at slaughter and matured in vitro for 24h in TCM-199 supplemented with 10% foetal calf serum, 10 microg ml(-1) FSH and LH, and 1microg ml(-1) of oestradiol. Fertilisation medium was IVF-SOF modified to contain 5mM Ca(2+) and no glucose. Experiment 1 investigated the addition of heparin, BSA (8 mg ml(-1)) or 20% SS. All oocytes were penetrated when IVF-SOF was supplemented with SS compared to 10 and 0% penetration when either heparin or BSA was present (P<0.01). However, 43.8% of these oocytes were polyspermic when the medium contained SS. In Experiment 2, the effect of sperm concentration on penetration rates during in vitro fertilisation was investigated. Total sperm penetration and monospermic penetration rates increased with increased sperm concentrations in a log linear manner (P<0.001) and both approached an asymptote at 0.4 x 10(6) sperm ml(-1) with 93.6 and 77% for total and monospermic penetration, respectively. Polyspermic fertilisation also increased with increasing sperm concentrations (P<0.05) but was variable (range 3.5+/-4.2 to 42.3+/-10.6%), especially at the lower sperm concentrations. Experiment 3 investigated the viability of these oocytes after transfer into red deer recipients. Fifteen 2- and 4-cell embryos were transferred into the oviducts of synchronized recipients 28 h post in vitro insemination. An additional fourteen embryos (8-10 cell) were transferred into synchronised recipients after 48 h of in vitro culture in either SOFaaBSA (n=10) or on red deer epithelial oviduct monolayers (n=4). Five (33% 5/15) of the recipients that received 2- and 4-cell embryos were pregnant at Day 45 (verified by ultrasonography) and four recipients subsequently calved. One recipient receiving an embryo cultured in SOFaaBSA was pregnant at Day 45 and subsequently calved. The birth of five normal calves indicate that full developmental competence of red deer oocytes matured and fertilised in vitro can be achieved by the techniques described.

A comparison of biochemical and meat quality variables in red deer (Cervus elaphus) following either slaughter at pasture or killing at a deer slaughter plant.

To investigate effects of pre-slaughter handling on blood and muscle biochemistry and venison quality, paddock-shot (n=8) and commercially slaughtered red deer (n=8) were compared. The deer were kept in two larger groups. One stag per group per day was head-shot, exsanguinated, electrically stimulated then transported 150 m to a deer slaughter premises (DSP) for processing. Prior to each slaughter day one of the groups was mustered into a deer yards and six (including two experimental) deer were selected for commercial handling and processing (including electrical stimulation) at the same DSP. Blood samples taken during exsanguination showed higher levels of cortisol, progesterone, glucose, lactate, albumin, creatine kinase, lactate dehydrogenase, aspartate aminotransferase and packed cell volume in the DSP-killed deer compared with the paddock-shot deer (P<0.05). Interpretation of these values indicated that pre-slaughter handling created moderate stress and high levels of muscular exertion or damage, possibly related to antagonism during lairage. However muscle glycogen, pH and meat quality measurements showed only minor, muscle-specific differences between treatments.

Storage of semen and artificial insemination in deer.

Methods of collection and freezing of semen of some deer species and aspects of controlled reproduction associated with the use of frozen-thawed semen by artificial insemination (AI) are discussed.

Genetic influences on reproduction of female red deer (Cervus elaphus) (1) seasonal luteal cyclicity.

This study compared the onset and duration of the breeding season of female red deer (Cervus elaphus scoticus) and its hybrids with either wapiti (Cervus elaphus nelsoni) or Père David's (PD) deer (Elaphurus davidianus). In Trial 1 (1995), adult red deer (n=9), F1 hybrid wapiti x red deer (n=6) and maternal backcross hybrid PD deer x red deer (i.e., 14 PD; n=9) were maintained together in the presence of a vasectomised red deer stag for 12 months. They were blood-sampled daily or three times weekly so that concentration profiles of plasma progesterone could be used to identify the initiation, duration and cessation of luteal events. There was clear evidence of luteal cyclicity between April and September, with the transition into breeding associated with an apparent silent ovulation and short-lived corpus luteum (i.e., 6-12 days) in every hind. A significant genotype effect occurred in the mean time to first oestrus (P<0.05), with wapiti hybrids and 14 PD hybrids being 9 and 5 days earlier than red deer. Between six and nine oestrous cycles were exhibited by each hind, with no difference in mean cycle length (19.5-19.6 days) between genotypes (P0.10). The overall length of the breeding season was significantly longer for wapiti hybrids (143 days) than for either red deer (130 days) and 14 PD hybrids (132 days, P<0.05). In Trial 2 (1998), adult red deer (n=5), 14 PD hybrids (n=5) and F(1) PD x red deer hybrid (n=5) hinds were maintained together from mid-February (late anoestrus) to early May, in the presence of a fertile red deer stag from 1 April. Thrice-weekly blood sampling yielded plasma progesterone profiles indicative of the onset of the breeding season. Again, there was a significant genotype effect on the mean time to first oestrus (P<0. 05), with F(1) PD hybrids and 14 PD hybrids being 13 and 5 days earlier than red deer. However, conception dates were influenced by the timing of stag joining, and were not significantly different between genotypes. The results indicate genetic effects on reproductive seasonality. However, seasonality observed for PD x red deer hybrids more closely approximated that of red deer than PD deer.